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Skylab Experiments, Volume 4, Life Sciences.National Aeronautics and Space Administration,Washington, D.C.EP-113May 73104p.; For related documents, see SE 016 464 and 465,SE 017 106, and SE 017 788-790Superintendent of Documents, Government PrintingOffice, Washington, D.C. 20402

EDRS PRICE MF-$0.75 HC-$5.40 PLUS POSTAGEDESCRIPTORS *Aerospace Technology; *Biological Sciences; Biology;

*Instructional Materials; *Science Activities;Science Education; *Science Materials; SecondarySchool Science

IDENTIFIERS NASA; *Skylab Education Program

ABSTRACTBasic knowledge about Skylab experiments is presented

in this book, one of a series, for the purpose of informing highschool teachers about scientific research performed in orbit andenabling the teachers to broaden their basis for material selection.This fourth volume is concerned with experiments designed to improveman's understanding of himself and his physiological functions andneeds, and studies on the biochemical and biophysical behavior oflower organisms and single human cells in the weightless environmentof space. The content includes topics relating to mineral balance,bioassay of body fluids, bone mineral measurement, cytogenic studies,Ian's immunity during and following exposure to space flight, blood-volume and red cell life span, red blood cell-metabolism, hematologiceffects, inflight lower body negative pressure, vectorcardiogram,metabolic activity, human vestibular function, sleep monitoring,effects of zero gravity on single human cells, and experiments withpocket mice and vinegar gnats. Experiment background, scientificobjectives, relationships between the Skylab investigations and highschool topics, and suggestions for classroom activities are provided.Included is a glossary of terms. (CC)

EP- i'31 1 ljt_A-.4k 1 T.,t. N T OF MLAL TH

1.1 01:LIU( :al ION

ExperimentsVolume 4Life Sciences

Information for Teachers, Including Suggestionsc on Relevance to School Curricula.

NATIONAL AERONAUTICS AND SPACE ADMINISTRATION

ri.

Experiments

Volume 4Life Sciences

Produced by the Skylab Program and NASA's Education ProgramsDivisin.n in Cooperation with the University of Colorado

NATIONAL AERONAUTICS AND SPACE ADMINISTRATIONWashington, D.C. 20546, May 1973

For sale by the Superintendent of Documents, U.B. Government Printing Office, Washington, D.C. 20402

PREFACE

Characteristically, new scientific knowledge reaches general application in classrooms yearsafter it has been obtained. This long delay stems, to a large extent, from a lack of awarenessthat information is available and that it has relevance to secondary school curricula. Toaccelerate this process, the National Aeronautics and Space Administration has prepared aseries of documents concerning Skylab experiments to apprise the educational communityin detail of the investigations being conducted in the Skylab Program, and the types ofinformation being produced.

The objective is not to introduce the Skylab Program as a subject in the classroom, butrather to make certain that the educational community is aware of the information beinggenerated and that it will be available for use. Readers are urged to use these books as anaid in planning development of future curriculum supplement material to make the mostappropriate use of this source of scientific knowledge.

National Aeronautics and Space AdministrationWashington, D.C. 20546

May 1973

11

CONTENTS

SECTION 1INTRODUCTION 1

SECTION 2MINERAL AND HORMONAL BALANCE 5Skylab Experiment M071, Mineral Balance 11Skylab Experiment M073, Bioassay of Body Fluids 13Skylab Experiment M078, Bone Mineral Measurement 15

SECTION 3HEMATOLOGY AND IMMUNOLOGY 17Skylab Experiment M111, Cytogenic Studies of the Blood 23Skylab Experiment M112, Man's Immunity, in vitro Aspects 25Skylab Experiment M113, Blood Volume and Red Cell Life Span 27Skylab Experiment M114, Red Blood Cell-Metabolism 29Skylab Experiment M115, Speeial Hematologic Effect 31

SECTION 4CARDIOVASCULAR STATUS 35Skylab Experiment M092, Inflight Lower Body Negative Pressure (LBNP) 41Skylab Experiment M093, Vectorcarcliogram 43

SECTION 5ENERGY EXPENDITURE 47Skylab Experiment M171, Metabolic Activity 49

SECTION 6NEUROPHYSIOLOGY 53Skylab Experiment M131, Human Vestibular Function 53Skylab Experiment, Sleep Monitoring 59

SECTION 7BIOLOGY 65Skylab Experiment 5015, Effects of Zero Gravity on Single Human Cells 66Skylab Experiment S071,Circadian RhythmPocket Mice 69Circadian RhythmVinegar Gnats 72

SECTION 8CLASSROOM ACTIVITIES 77

SECTION 9GLOSSARY OF TERMS 87

iii

INTRODUCTION

The Skylab Education Program

This year the United States' first manned scientific space station, Skylab, was launched intoorbit to be the facility in which successive crews of astronauts can perform more than 270scientific investigations in a variety of fields of interest. These investigations can be dividedinto four categories: physical sciences, biomedical sciences, earth applications, and spaceapplications.

The Skylab Program will produce information that will enhance present scientific knowledgeand perhaps extend the frontiers of knowledge on subjects ranging from the nature of theuniverse to the structure of the single human cell. It is the objective of the NationalAeronautics and Space Administration that the knowledge derived from the SkylabProgram's investigations be made available to the educational community for applications tohigh school education at the earliest possible date.

For this reason, the Skylab Education Program was created to assure that maximumeducational benefits are obtained from the Skylab effort, documentation of Skylabactivities is adequately conducted, and understanding of scientific developments isenhanced.

This document, one of several volumes prepared as part of the Skylab Education Program,has the dual purpose of (1) informing high school teachers about the scientific investigationsperformed in Skylab, and (2) enabling teachers to evaluate the educational benefits theSkylab Program can provide.

These books will define the objectives of each experiment, describe the scientificbackground on which the experiment is based, outline the experimental procedures, andindicate the types of data anticipated.

In preparing these documents an attempt has been made to illustrate relationships betweenthe planned Skylab investigations and high school science topics. Concepts for classroomactivities have been included that use specific elements of Skylab science as focal points fordemonstrations of selected subjects. In some areas these address current curriculum topicsby providing practical applications of relatively familiar, but sometimes abstract principles;in other areas the goal is to provide an introduction to phenomena rarely addressed in highschool science curricula.

It is the hope of the National Aeronautics and Space Administration that these volumes willassist the high school teacher in recognizing the educational value of the informationresulting from the Skylab Program which is available to all who desire to make use of it.

Application

Readers are asked to evaluate the investigations described herein in terms of the scientificsubjects taught in secondary schools. The related curriculum topics identified should serveas suggestions for the application of Skylab Program-generated information to classroomactivities. As information becomes available from the Skylab Program, announcements willbe distributed to members of the educational community on the NASA EducationalPrograms Division mailing list. To obtain these announcements send name, title, and fullschool mailing list (including zip code) to:

iv

National Aeronautics and Space AdministrationWashington, D.C. 20546

Mail Code FE

This volume covers a broad spectrum of scientific investigations on Skylab that have beendesigned to improve man's understanding of himself and his physiological functions andneeds. Additionally, a small group of related studies on the biochemical and biophysicalbehavior of lower organisms and single human cells in the weightless environment of space isincluded.

So that the reader will have a better understanding of the timing, setting, and scope of thetotal scientific endeavor on Skylab, a brief description of the Skylab Program has beenincluded.

Acknowledgments

Valuable guidance was provided in the area of relevance to high school curricula by Dr.James R. Wailes, Professor of Science Education, School of Education, University ofColorado; assisted by Mr. Kenneth C. Jacknicke, Research Associate on leave from theUniversity of Alberta, Edmonton, Alberta, Canada; Mr. Russel Yeany, Jr., ResearchAssociate, on leave from the Armstrong School District, Pennsylvania; and Dr. Harry Herzerand Mr. Duane Houston, Education and Research Foundation, Oklahoma State University.

The Skylab Program

The Skylab orbiting space station will serve as a workshop and living quarters for astronautsas they perform investigations in the following broad categories: physical sciences,biomedical sciences, Earth applications, and space applications.

The spacecraft will remain operational for an eight-month period, manned on threeoccasions and unmanned during intervening periods of operation. Each manned flight willhave a crew of three different astronauts. The three flights are planned for durations of onemonth, two months, and two months, respectively.

A summary of objectives of each of the categories of investigation follows.

Physical Science

Observations free of filtering and obscuring effects of the Earth's atmosphere will beperformed to increase man's knowledge of (1) the sun and of its importance to Earth andmankind, and (2) the radiation and particulate environment in near-Earth space and thesources from which these phenomena emanate.

Biomedical Science

Observations under conditions different from those on Earth will be made to increase man'sknowledge of the biological functions of living organisms, and of the capabilities of man tolive and work for prolonged periods in the orbital environment.

Earth Applications

Techniques will be developed for observing from space and interpreting (1) Earthphenomena in the areas of agriculture, forestry, geology, geography, air and water pollution,land use and meteorology, and (2) the influence of man on these elements.

Space Applications

Techniques for adapting to and using the unique properties of space flight will be developed.

The Skylab Spacecraft

The Skylab cluster contains five modules (see illustration).

1) The orbital workshop is the prime living and working area for the Skylab crews. Itcontains living and sleeping quarters, food preparation and eating areas, and personalhygiene equipment. It also contains the equipment for the biomedical science experimentsand for some of the physical science and space applications experiments. Solar arrays forgeneration of electrical power are mounted outside this module.

2) The airlock module contains the airlock through which suited astronauts emerge toperform activities outside the cluster. It also contains equipment used to control thecluster's internal environment and the workshop electrical power and communicationssystems.

3) The multiple docking adapter provides the docking port for the arriving and departingcommand and service modules, and contains the control center for the telescope mountexperiments and systems. It also houses the Earth applications experiments and materialsscience and technology experiments.

4) The Apollo telescope mount houses a sophisticated solar observatory having eighttelescopes observing varying wavelengths from visible, through near and far ultraviolet, toX-ray. It contains the gyroscopes and computers by which the flight attitude of Skylab iscontrolled. Solar arrays mounted on this module generate about half of the electrical poweravailable to the cluster.

5) The command and service module is the vehicle in which the crew travels from Earthto Skylab and back to Earth, and in which supplies are conveyed to Skylab, and experimentspecimens and film are returned to Earth.

Skylab will fly in a circular orbit about 436 kilometers (235 nautical miles) above thesurface of the Earth, and is planned to pass over any given point within latitudes 50° northand 50° south of the equator every five days. In its orbital configuration, Skylab will weighover 91,000 kilograms (200,000 pounds) and will contain nearly 370 cubic meters (13,000cubic feet) for work and living space (about the size of a three bedroom house).

vi

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1 Apollo telescope mount 6 Ward room 10 Airlock module2 Solar arrays 7 Orbital workshop 11 Multiple docking adapter3 Sleeping quarters 8 Experiment compartment 12 Earth resources experiments4 Personal hygiene 9 Airlock external hatch 13 Command and service module5 Biomedical science experiment

Figure 1 Skylab Orbiting Station

Skylab Life Science Investigations

Major study emphasis in the Skylab Program is on life sciences as summarized in Figure 2.

The human body incorporates a large number of interdependent body systems inhomeostatic balance, which is essential to the proper functioning and well being of the bodyas a whole. Therefore, the program shown in figure 2 recognizes that measurement ofchange in a single body system or function is not a final answer in itself. Ultimate answersrequire a considerable amount of cross correlation among the six principal study areas.

The six areas of scientific investigations being emphasized in Skylab appear on the left sideof the figure. The investigative program for these areas includes measurement of individualevents and subsequent integration of all data to establish the effects of interactions.

The inquiring reader of this book can discover that the educational potential of the programof investigations discussed is not limited to the specific scientific thrust of the experimentbut has a broader association which cuts across many elements of the high schoolcurriculum. Thus, the general subject area of this book, life sciences, can provide sourcematerial in such diverse fields as physics, biology, and chemistry.

Table 1 is a cross index of general curriculum elements to specific investigations within theSkylab Life Sciences Experiment program.

vii

Mineral and HormonalBalance

Hematology andImmunology

M071 Mineral Balance

M078 Bone MineralMeasurement

M073 Bioassy ofBody Fluids

Cardiovascular Status

Energy Expenditure

Neurophysiology

Biology

M092 Lower BodyNegative Pressure

M093 Vectocardiogram

M171 Metabolic7Activity

.....1

M111 CytogeneticStudies of Blood

..4,HM112 Man's Immunityin vitro Aspects

LM113 Blood VolumeRed Cell Life Span

H M114 Red BloodCell Metabolism

5015 Effects of Zerogon Single Human Cells

Figure 2. Experimental Program

viii

5071 Circadian Rhythm,Pocket Mice

.0....i S072 Circadian Rhythm,Vinegar Gnat

M115 SpecialHematologic Effects

M131 HumanVestibular Function

M133 SleepMonitoring

Tab

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Section 1

Introduction

MannedSpaceflight

EXPERIMENTS PROGRAM BACKGROUND

Since the beginning of manned spaceflight in the UnitedStates, there has been a continuing concern regarding man'sability to live and operate efficiently while on extendedflights in space. Answers to this concern can be obtained onlythrough a careful quantitative assessment of the crews'performance and physiological well being while working inthe space environment. The study must develop data ontrends and rates of adjustments while providing aninstantaneous assessment of the crewmen's status andperformance at critical points during the flight. Equallyimportant is the ability of the crew to readjust to life onEarth after return from orbit. Interpretation of data dependson comparing flight results with a data base obtained inground laboratories. Such a combination of ground andinflight experimentation forms the basic plan for the Skylabbiomedical program.

Missions up to 14 days have produced changes in theastronaut's body tissues and systems as expected. Althoughthe vector of these changes has been in the expecteddirection, individual changes have often differed from thepredicted value. While these earlier missions have shown thatman's gross capabilities to operate in the space environmentcontinue at or near Earth equivalence, new techniques forperforming a given task on orbit have sometimes beenrequired to reflect the unique character of the spaceenvironment.

The Skylab Program and a life science experiments that areto be performed offer an opportunity to increase man'sknowledge of the biological and physiological functions ofliving organisms. This will be achieved by making scientificobservations of living organisms in an environment differentfrom that on Earth, and evaluating the transitional effects asthe organism responds and hopefully adapts to these newconditions.

Specifically, the Skylab biomedical experiments provide Data requirements for longinformation on man's ability to-- duration spaceflight planning

1) perform effectively while in near weightless flight up to56 days,

2) identify natural biological cycles,

3) identify the adequacy of or need for additionallife-support provisions to control changes in bodychemistry within acceptable bounds,

4) confirm or refine the habitability design criteria forfuture long-duration manned systems.

1

PhysiologicandPsycho logicChanges

2

In addition, information will be developed to aid inunderstanding crew psychological reactions to long-durationconfinement; evaluating the problems associated with food,water, and waste management; and for developing criteria fordesigning better crew-monitoring systems for future missions.The Skylab Program should provide these data at asufficiently early date to affect the next major mannedsystems.

In terms of life-science research, Skylab aims primarily atidentifying and defining physiologic and psychologic adaptivechanges and establishing whether these will be progressive orself-limiting. If progressive, they could necessitate returning acrew back to Earth before the scheduled time. If self-limiting,the crew should pass through a period of adjustmentfollowed by stabilization appropriate to the newenvironment. It is well known that when man travels to ahigh-altitude city or works in an undersea environment forlong periods, the body processes pass through an initialadjustment followed by stabilization at new levelsappropriate to survival in the new environment. Thistransient adjustment is also expected for the Skylab crew.The new stable levels of accommodation are expected to beadequate for the normal health and function of man duringcontinued orbital flight. Upon return to the terrestrialenvironment the space-stabilized levels of accommodationmay become significant factors in the crews' well being; thatis, the acclimatization to space which has occurred mayrequire specialized assistance to help the crewmen readapt toEarth following long duration space flight. Based on shorterduration past missions, the predicted course of events isencouraging for these longer manned spaceflights.

The experiments selected for Skylab reflect a synthesis of theobservations made during previous flights. Body systemswhich have shown changes and which could present aproblem for the crewmen have received priority in thiscontinuing research.

These studies have been organized for the purposes of thisdocument in the following order:

Section 2, Mineral and Hormonal Balance

Section 3, Hematology and Immunology

Section 4, Cardiovascular Status

Section 5, Energy Expenditure

Section 6, Neurophysiology

Section 7, Biology

Acclimatable and accommo-dative reflexes trigger transi-tional adjustments in bodysystems.

In each section a general introductory background isincluded. The purpose of this background is to establish afundamental relationship between the experimental programand basic classroom science. This background is orientedtowards establishing a broad scientific link between theSkylab ex perimental program and classroom sciencecurricula. Hence, this approach emphasizes the generalscientific background of the experimental program andseparates the Skylab technological goals from the broaderteaching aspects. Other subsections describe the specifictechnical objectives, equipment, and expected data outputfrom each experiment.

3

Section 2Mineral andHormonal BalanceMineral BalanceSkylab Experiment M071

Bioassy of Body FluidsSkylab Experiment MG; 3

Bone Mineral MeasurementSkylab Experiment M078

CN

0

U(1)

Man's In order to maintain life, an organism must be able toCommunication coordinate a wide variety of different functions. For this

purpose, animals, including man, have developed twoimportant communication systems. The nervous system hasthe form of an elaborate telegraph (neural) network in whichcommunications paths are reasonably well defined byanatomical connections which, in many cases, provide veryspecific routing of transmitted messages to discreteanatomical sites. The other communication system is theendocrine system consisting of many ductless glands (Figure2-1). In it, messages are carried in the bloodstream in theform of complex chemical substances (hormones) thatinteract with special cells to receive the hormone and to reactto it in a specific way. These "target cells" may be limited toone particular type of cell, e.g., the gonadal cells that respondto the gonadotrophic hormone, or the renal tubular cells thatrespond to aldosterone. However, some hormones, such asinsulin, act upon many different types of cells.

System

.4Brain

Hypothalamic Nuclei

Hypophysis (Pituitary)

Thyroid

Parathyroids

Pancreas

Adrenal Cortex

Adrenal Medulla

Kidney

Testes (in male)Ovaries (in female)

Figure 2-1 Major Endocrine Glands in Man

Hormonesany of variousinternally secreted compoundsformed in endocrine organs thataffect the functions of receptiveorgans when carried to them bybody fluids.

5

Neuroendo-crine System

6

There are many interactions between the nervous andendocrine systems, so that they are sometimes consideredcollectively as the neuroendocrine system. For example,transmission between neurones in the sympathetic nervoussystem occurs via adrenergic hormones. These hormones are Adrenergicactivated oralso produced by the adrenal medulla, an organ which transmitted by epinephrine.functions as an endocrine gland, but is embryologicallyderived from neural tissue. Another example of the closerelationship between the neural and endocrine systems is therequirement that certain hormones be present for the normaldevelopment and functioning of nerve tissue. It is known, forinstance, that lack of thyroid hormone in infancy results inmental retardation, and in adults in the slowing ofconduction in the nerves. The complex systems whichcontrol some physiological variables such as body watercontent are known to include both neural and endocrineelements. The type of action which hormones may elicitfrom target cells is shown in Table 2-1. The extent ofinteractions between the neurohormonal processes and thebasic behavior of target cells is shown in Figure 2-2.

Table 2-1 Hormone Elicited Control

Altered blood flow to cells, e.g., nearly all tropichormones on target glands.Altered membrane properties of specific cells.

1) ADH: renal loop of Hen le and H2 0 transport.2) Aldosterone: renal proximal tubule and Na

transport.3) Various hormones and amino acid transport, e.g.,

178-estradial increases transport by uterus.4) Insulin: glucose transport; K' transport; Pi

transport; and transport into muscles but notliver.

Altered rates of protein metabolism.1) Anabolismincreased protein synthesis e.g.,

a) GH: bone, muscle, liverb) TSH: thyroid; ACTH; adrenal cortex, etc.c) Thyroxin: many cellsd) Cortisol: liver

2) Catabolismincreased protein degradationa) Cortisol: muscle, boneb) Thyroxin (large doses): many cells

Altered enzymatic activity1) CHO Metabolism e.g.,

a) Epinephrine: liver, muscle phosphorylaseand glycogenolysis

b) Insulin: muscle and glycogen synthesis2) Lipid metabolism

a) Many hormones: lipolysis in adiposetissue.

b) Insulin: antilipolysis in adipose tissue.Mineral, water and electrolyte metabolismEffects may be secondary to those listed in categoriesabove, e.g., PTH and Ca4÷ mobilization may besecondary to increased catabolism of bone protein, oraltered membrane changes.

Sensory Input

Change in External ofInternal Environment

Brain

Portal Circulation

Concentrationof Componentin Plasma

R F

ETC

Parathyroid

ADH Oxytocin

ACTH

AdrenalCortex

CNS

AdrenalMedulla

Epinephrine

Cortisol;Aldosterone

TSH

Thyroid

ThyroxinTRIT

MSH GH PTH

Estrogens Androgens

Glucose

Pancreas

Insulin

Relatively Hormone Specific Target Cells

Output = Alteration in Metabolic Behavior of Target Cells

Figure 2-2 Neurohumoral Interactive Processes Influencing Metabolic Activity of Specific Cells

7

Compositionof Bone

8

Referring to Figure 2-2, it is apparent that the interactionsare many and complex and further that these actions can betriggered by outside environmental influences that can affectcomponent concentration in the plasma, the portalcirculation, and even the brain through sensory organ inputs.During the evolution of man, gravitational forces haveunquestionably played a major role in his physiologicaldevelopment. The significance of this force in the health andgrowth of man is of uncertain limits today.

One rather interesting aspect of this is demonstrated by theinfluence which such external forces may have on the growthof bone in the body.

Bone is a unique tissue comprising a very special body organsystem. Living bone requires the same nutrients, controls,and environments as other living tissue in the body. Incontrast, however, to other body tissue and because of itscalcified nature, bone provides a semipermanent biologicalrecord. Hence, like the growing tree, the structure of bonerecords the anomalies of its past including irregularities ofdiet, disease, and environment.

Bone shows many levels of organization (Fig. 2-3). Typically,hones are composed of two basic types of material: compact(iamellar) bone and spongy (cancellous) bone. Compact bonemakes up the wall of the central portion (diaphysis) as well asthe outer shell of the flared ends (metaphysis). The interiorof the metaphysis is composed of spongy bone which in timeis built up of a collection of plates and bars called trabeculae(little beams). There is little difference in the bony materialin these two types of bone although the porosity of eachtype is quite different.

The outer surface of the bone is covered by a membraneknown as the periosteum: the inner surface is covered by amembrane known as the endosteum. The volume that iscontained within the wall of the diaphysis is called themedullary cavity and it is filled with marrow as are the spacesbetween the trabeculae of spongy bone. Yellow marrow isfound in the large cavities of the long bones. It consists forthe most part of fat cells. It may be replaced by red marrowin anemia. Red marrow is the site for production of thegranular leucocytes and the erythrocytes (red blood cells).Red marrow is found in the flat and short bones, the articularends of the bones, the bones of the vertebrae, the cranialdipole and the ribs. Both types of marrow have a supportingconnective tissue and numerous blood vessels.

Bone is formed by cells called osteoblasts, and destroyed orresorbed by other cells called osteoclasts. These two celltypes are formed from undifferentiated mesenchymal cellslying on the surface and in the spaces of the bones. When a

Cancellous of a reticular,spongy, or lattice-like structure:used mainly of bony tissue.

Diaphysisthe shaft of a longbone.

Metaphysisthe wider part atthe extremity of the shaft of along bore, adjacent to theepiphysical disc.

BoneStructure

Epiphyses

Spongy bone

Articulates withAcetabulum

Compact bone

Periosteum

YellowMarrow

NutrientArtery

MedullaryCavity

--.,011111111111

HaversianCanal

Line of EpiphysealFusion

Articulate withPetalis and Tibia

Figure 2-3 Structure of Human Femur

Osteocyte

Canaliculi

ConcentricLamellae

CircumferentialLamellae

certain stimulus is received, these mesenchymal cells beginfunctional and morphologic transitions which terminate inthe formation of specilized osteoblasts and osteoclasts asappropriate to the stimulus. The deposition or mobilizationof calcium under the influence of structural loads on thebone completes the growth of new adult bone. Bone cells(osteocytes) were originally osteoblasts which becameincluded in the bone matrix. They are the living elements ofbone tissue and their function is to maintain the chemicalenvironment of the surrounding nonliving tissue. In lamellarbone, there are about 20,000 osteocytes per cubic millimeter,residing in the same number of lacunae (spaces) and they areabout 10 x 15 x 25 microns in size. Osteocytes die for avariety of reasons; after death they are absorbed and leaveempty lacunae behind. Human osteocytes live an average of25 years.

There are a wide variety of observations that indicate that thegrowth system, whose function is to penetrate and remodel

Osteoblasta cell that arisesfrom a fibroblast and which, asit matures, is associated with theproduction of bone.

F ibroblasta connective tissuecell.

Osteocytesan osteoblast thathas become imbedded within thebone matrix, which connects toother osteocytes to form bone.

9

BoneMeasurement

10

bone, is somehow responsive to the local state of stress; thatis, physical loads of various kinds alter the growth rate,shape, and structure of bone.

Measurements of bone weight show that inactivity throughconfinement, immobilization, or paralysis causes a significantreduction in both the amount and quality of bone. Severalstudies of this phenomena involving patients immobilized inpelvic girdle and leg casts have been made. During theconfinement period, significant levels of calcium,phosphorus, and nitrogen were found in urinary and fecalexcretions. Animal experiments indicate that immobilizationof a limb invariably leads to disuse atrophy and a reductionin muscle and bone weight.

Long bones with an angulation caused by a poorly set but Angulationthe formation of ahealed fracture have been observed in children to gradually sharp obstructive anglestraighten. These changes and adjustments in shape requirebone to be resorbed on the tension or minimum compressionside and deposited on the more heavily loaded side. Thus, itcan be observed that the bone cross section drifts in adirection to minimize loading eccentricity so that boneappears to possess a stress directed growth regulatingmechanism which adjusts the bone architecture in responseto external loads.

Generally, it can be said that usage in the young growingskeleton leads to changes of shape and structure while inolder, fully grown bones usage leads mainly to changes instructure. It is as if the growing bone would grow into a newpattern, determined by mechanical forces, while fully grownbone has become more rigid in its outer shape, but can adaptto changed forces by a new orientation of its internalstructural elements.

The skeleton during man's daily activities is exposed to awide range of loads varying in magnitude and direction. Thegrowth stimulus will therefore also vary in both intensity anddirection. Thus, bone organization in the adult is finallydetermined by those activity patterns most frequent in thelifetime of the individual, superimposed, of course, on thegenetically controlled pattern unique to the species. Activitypatterns not characteristic of the species would therefore beexpected to stimulate growth contrary to genetically carriedtraits, and result in a bone weakened for characteristicactivities of the species.

The Skylab Program affords a unique near-weightlessenvironment in which to test many of the theories that havebeen advanced as to the manner in which body health andgrowth are influenced by the environment, and specifically toevaluate the role of mineral balance and neurohumoralinfluence on this growth.

MineralBalance

SKYLAB EXPERIMENT M071

Bed rest immobilization studies have shown that in healthyyoung adults urinary calcium increases to 2-3 times thecontrol level within 5 weeks after confinement. X-ray studiesof the bones have demonstrated demineralization as early as2-3 weeks after immobilization. Gemini pre- and postflightx-rays have suggested a similar loss of mineral from peripheralbones; the Gemini 7 mineral balance experiment hasdemonstrated a trend toward negative mineral balance.Continuous losses of calcium and nitrogen, such as thoseduring long duration missions, might result in impairment ofskeletal and muscle integrity and the formation of kidneystones. Identification of the rates of actual deterioration willallow specific countermeasures to be taken on later flights,such as the institution of exercise routines and themanipulation of dietary constituents.

The principal method of assessing the effect of a stressor onthe biochemical integrity of the skeletal and muscularsystems is to determine whether the stressor promotes acatabolic response that is greater than the anaboliccapabilities of the tissues. The change in equilibrium may bereflected in an imbalance between the nutrient intake of theconstituent in question and the output of it and/or itsmetabolites. A state of negative nitrogen or calcium balanceis not itself detrimental unless it is of an extent and durationthat results in compromise of the integrity of muscle or bonewith resultant increases in susceptibility to disease or actualpathology. Prior to the onset of recognizable disease,however, minor changes in function suggesting laterdeterioration can be demonstrated.

SCIENTIFIC OBJECTIVES

The objective of this experiment is to determine the effectsof space flight on the muscle and skeletal body systems byquantitative assessment of the gains and losses of biochemicalconstituents of metabolic importance. These constituents arewater, calcium, phosphorus, magnesium, sodium, potassium,nitrogen, urea, hydroxyproline, creatinine and chloride.

EQUIPMENT

The Mineral Balance and Bioassay of Body Fluids experimentdata were obtained from a detailed, daily inventory of foodand water intake, of body mass, and of output of wasteproducts. Waste products, feces, urine and vomitus aremeasured, processed, and stored onboard for return to Earthwith the crew, and subjected to detailed analysis. Equipmentdesigned to perform the specimen and body massmeasurement, M074 and M0172, respectively, is used tosupport this experiment.

Stressora stimulus, such as painor fear, that disturbs orinterferes with the normalphysiological equilibrium of anorganism.

Anabolismany constructiveprocess by which samplesubstances are converted intomore complex compounds,constructive metabolism.

Catabolismany destructiveprocess by which complexsubstances- are converted byliving cells into more simplecompounds; destructivemetabolism.

11

ExperimentInvestigations

12

PERFORMANCE

The performance of the Mineral Balance Investigations starts21 days before flight and continues throughout the flight andfor 18 consecutive days after return to Earth. Each day ofthe observing period, the following functions will beperformed by the crew:

1) Body weight (or mass) will be measured immediatelyafter the first urine voiding following the sleep period.

2) A predetermined diet will be used since the compositionof the crewman's diet must be known and carefullycontrolled. In the preflight period, each crewman will usethis diet to allow the establishment of individualmetabolic equilibrium. Every effort will be made to makethe diet palatable. The premeasured menu for each mealwill be prepared and the mass of any leftover food will bemeasured and recorded.

3) Fluid can be taken as desired, but all intake will berecorded. This includes fluid used for foodreconstitution.

4) All urine, feces, and vomitus will be collected pre- andpostflight and preserved for analysis. Inflight, the amountof daily urine output from each crewman will bedetermined, and a measured homogeneous sample of atleast 45 milliliters (2 for tracer method volumedetermination) taken, frozen, and stored for analysis. Allfeces and vomitus passed will he collected, the mass willbe measured, and the specimens will be dried and storedfor postflight analysis.

5) Periodic blood samples will be taken and theconcentration of selected constituents determined.Inflight blood samples will be processed and frozen forpostflight analysis.

DATA

During the Skylab Program, three crews (three men in each)will occupy the orbital workshop on three separate occasions.The initial mission will last up to 28 days and the other twofor up to 56 days each. The Mineral Balance Experiment willbe performed on all three missions so that by the end of theSkylab Program a continuous quantitative assessment of themuscle and skeletal body systems for nine differentindividuals will have been obtained. For each individual, apreflight baseline will be followed by a day-by-day profile ofhis physiological reaction to the space environment, andpostflight, his readaptation to Earth's normal conditions.Specifically, the following data on a daily basis will beobtained preflight, inflight and postflight:

Bioassy ofBody Fluids

1) food consumptionnutritional, mineral, and caloriccontent;

2) fluid consumption;3) fecesmass and concentration of biochemical

constituents;4) urinetotal voids volume and concentration of the

biochemical constituents;5) vomitusmass and concentration of the biochemical

constituents;6) body mass.

In addition, blood samples will be taken periodically pre-, in-and postflight. These samples will be analyzed to determinealkaline phosphates, total protein, electrophoresis pattern,sugar, calcium, phosphorus, magnesium, sodium, potassium,hydroxyproline, chloride, and creatinine.


Although many external influences contribute to theenvironment of the human organism as a whole, theenvironment of its basic unit, the living cell, is whollyinternal. Since changes in extracellular fluid produce changesin the composition of the intracellular fluid, it is essential tothe normal function of cells that the constancy of this fluidbe maintained. This is achieved by the close interaction ofseveral organ systems, the kidney holding a predominant role.The kidney is thus viewed as an organ which not onlyremoves metabolic wastes, but actually performs highlyimportant homeostatic functions by adjusting plasma volumeand composition.

The necessity of elucidating the homeostatic controlmechanisms that govern plasma volume and composition isevident when one realizes the complex and, as yet,unexplained interactions of these metabolic and endocrinecontrols. In the changing external environment, there is anarrow margin within which man's physiological well-beingcan be accommodated by these functions. Evidence exists tosuggest that these mechanisms may play a significant role inman's adaptation to stress, including gravity.


The objective of the Bioassay of Body Fluids Experiment isto evaluate the endocrinological adaptation resulting fromexposure to the spaceflight environment for periods up to 56days and to readaptation to the Earth environment.Specifically, the following elements in blood and/or urinewill be evaluated: adrenocorticotropic hormone (ACTH),17-hydroxy-corticosterone (Cortisol), angiotensin II, renin,

Homeostaticuniformity orstability in the normal bodystates of an organism.

13

ExperimentInvestigations

14

aldosterone, antidiuretic hormone (ADH), epinephrine,norepinephrine, urine electrolytes (sodium and potassium),urine and plasma osmolality, extracellular fluid volume, totalbody water, serum thyrocalcitonin parathyroid hormone,serum thyroxine, hydrocortisone, renin activity, total andfractional ketosteroids, insulin, human growth hormone, andthyroid stimulating hormone.

EQUIPMENT

The Mineral Balance and Bioassay of Body Fluids data isobtained from a detailed, daily inventory of food and waterintake, of body mass, and of output of waste products. Wasteproducts, feces, urine, and vomitus are measured, processed,and stored onboard for return to Earth with the crew, andsubjected to detailed analysis in the postflight period.Equipment for performing the specimen and body massmeasurements, M074 and M0172, respectively, is used tosupport this experiment.

PERFORMANCE

The experiments will be accomplished in three phases: (1)preflight, for 21 days, (2) inflight, and (3) postflight untilreadaptation has been established, beginning immediatelyafter flight.

The following functions will be performed each day of theobserving period:

1) Body weight (or mass) will be measured immediatelyafter the first urine voiding following the sleep period.

2) A predetermined diet will be used since the compositionof the crewman's diet must be known and carefullycontrolled. In the preflight period each crewman will usethis diet to establish individual metabolic equilibrium.Every effort will be made to make the diet palatable. Thepremeasured menu for each meal will be prepared and themass of any leftover food will be measured and recorded.

3) Fluid can be taken as desired, but all intake will berecorded. This includes fluid used for foodreconstitution.

4) All urine, feces, and vomitus will be collected pre- andpostflight and preserved for analysis. Inflight, the amountof daily urine output from each crewman will bedetermined, and a measured homogeneous sample of atleast 75 milliliters for bioassay of body fluidsexperiments taken, frozen, and stored for analysis. Allfeces and vomitus passed will be collected; the mass willbe measured; the specimens will be dried and stored forpostflight analysis.

Bioassydetermination of theactive power of a substance bytesting its effect in an organism.

Bone MineralMeasurement

5) Periodic blood samples will be taken and theconcentration of selected constituents determined.Inflight blood samples will be processed and frozen forpostflight analysis.

DATA

The Bioassay of Body Fluids Experiment will occur on allthree missions so that by the end of the Skylab Program, acontinuous quantitative assessment of the endocrinologicaladaptation for nine different individuals will have beenobtained. For each individual, a preflight baseline will beobtained, folIPwed by a day-by-day profile of hisphysiological reaction to the space environment, and afterflight, his rea riaptation to Earth normal conditions.Specifically, the following data on a daily basis will beobtained preflight, inflight, and postflight:

1) Data obtained from Mineral Balance, Experiment M0712) Urineconcentration of the biochemical constituents Biochemical -- dealing with living

specified in the Objectives. matter.

In addition, blood samples will be taken periodically pre-, in-and postflight and those parameters specified in theObjectives will be determined.


Stimulus of bone metabolism is a function of the forcesexerted on the bone by the attached muscles and the forceexerted along the longitudinal axis of the skeletal system bygravity. Both forces are altered during complete bed rest andabsence of gravity. Consequently, bone mineral losses havebeen associated with long-term bed rest and were anticipatedas a potential problem for the crews of long-term spaceflights.

In both Gemini and early Apollo flights, small but significantlosses have been measured in astronaut bone mass. Incontrast to the Gemini and early Apollo studies, which used aradiographic densitometry technique, the bone mineralstudies performed on Apollo 14, using the gamma rayabsorption technique, revealed no significant losses in bonemineral content. More data from both ground based andinflight studies are necessary to resolve the issue beforecommitting man to extended space travel.


The objective of the Bone Mineral Measurement experimentis to determine, by a photon absorptiometric technique, the

15

occurrence and degree of bone mineral changes in the Skylabcrewmen that might result from exposure to the weightlesscondition.

EQUIPMENT

The Bone Mineral Measurement experiment consists ofpreflight and postflight measurement of the condition of twobones in each astronaut. The ground based equipment forthis experiment is a photon scanning deyice in which aniodine-125 photon source is mounted in opposition to an Photon--a quantum of electro-x-ray detector. The astronaut's limb containing the bone to magnetic radiation.be measured is firmly held in the required position within thescanning device. Foot molds and restraining equipment areused for accurate positioning of the subject for the scan.

PERFORMANCE

Experiment The Bone Mineral Measurement performance using theInvestigations photon scanner starts 30 days before flight. The bones to be

measured are the heel bone (os-calcis) in the left foot and oneof the bones (radius) in the right forearm. (See Fig. 2-3.)X-rays of the heel will be made seven days before the firstphoton scanner observation.

Preflight scans of the left os-calcis and right radius will beaccomplished on the flight crew, backup crew, and on acontrol group 30, 14 and 3 days before the launch of theflight crew.

Postflight measurements will be made on the flight crew andon the control group within 10 hours after splashdown and atthe following intervals: 2 to :3 days, 5 tn 10 days, and 30 to45 days if baseline values have not been reached in the 5- to10-day period.

The measurement on recovery day should be made at theearliest time possible (within 10 hours) after crew recovery tominimize the effect of weight bearing by the os-calcis andmaximize the data accuracy on the extent of the bonemineral losses under zero gravity conditions.

Additional postflight measurements will be required if theflight crewmembers have not reestablished their baselinevalues by the third measurement.

DATA

The data returned by this experiment will be the pre- andpostflight bone density measurements of the os-calcis andradiqs of each Skylab crewmember (three from the 28-daymission and six from the 56-day mission) and nine controlgroup members. The data will be used to determine theimpact of the spaceflight environment on the degree andoccurrence of the bone mineral changes.

16

Section 3Hematology andIrnmunnlogy

Cytogenetic Studies of the Blood,Skylab Experiment M111

Man's "unity, !n Vitro Aspects,Skylab Experiment M112

Blood Volume and Red Cell Life Span,Skylab Experiment M113

Red Blood Cell Metabolism,Skylab Experiment M114

Special Hematologic Effect,Skylab Experiment M115

Hematology Hematology, the study of the form and function of blood, inlarge part is concerned with changes in the concentration ofthe functional elements of the blood. These changes cancome about as the result of disease processes andenvironmental changes, or they may be induced as apurposeful experiment.

Regardless of how these changes are precipitated, if theyexceed relatively narrow physiological limits they will beaccompanied by dramatic changes in the effectiveness of theindividual even to the extent of causing death. The followingdiscussion is intended to provide a brief resume of thefunctions and functional elements that form the basis for ourpresent understanding of the role of blood in health anddisease.

FUNCTIONS OF BLOOD

Every living organism, simple or complex, requires for itssurvival, the ability to exchange materials with itsenvironment, extracting from the environment thosematerials which it can metabolize for its energy requirementsand rejecting to the environment those materials whichwould poison it. In the case of single cell and other relativelysimple organisms, the process of transmembrane diffusionprovides an effective mechanism for satisfying this need.Larger, more complex (multicellular) organisms must providesophisticated organ systems to accomplish this function sincethe external environment makes intimate contact with theorganism's body only at its surface. For example, the skinsurface of the average adult human amounts to only about1.7 m2 resulting in a surface to volume ratio of about 0.02min2 /mm3 . Furthermore, this surface is relatively imperviousto most materials, and serves only as a means for eliminatingexcess heat and a small amount of water. Hence, theexchange of materials between the cells of the body and theexternal environment must be carried on by specializedsystems such as the lung, kidney, gastrointestinal (GI) tract,blood, and interstitial fluid.

These specialized systems are related as shown in the diagramof Figure 3-1. Reference to this diagram shows that while thelung, kidney, and GI tract interface with the externalenvironment, they do not interface directly with the cells ofthe body. The materials upon which the body is dependentpass through two intermediate mechanisms, the circulatorysystem, and the interstitial fluid. The interstitial fluid, whichbathes all cells in the body, provides a uniform environmentfrom which the cells extract their material needs and towhich they reject waste products. The circulatory systemprovides a transport mechanism that assures physical andchemical uniformity (within relatively narrow physiological

Interstitial Fluidthat portionof the body water outside of thecells and outside of the plasmavolume.

17

VitalTransportFunctions

18

limits) of the interstitial fluid. The circulatory system sopermeates the interstital compartment that in the heart, forinstance, no cell is further than about 0.008 mm from acapillary.

In addition to the transport of gases, material, and wasteproducts, blood has other equally vital functions. Thesefunctions include carrying the heat generated by the cells'metabolic activity to the surface for easy transfer to thesurrounding environment, and certain defense mechanisms bywhich bacteria and foreign particles are removed from thebody.

External Environment Interstitial fluid

Figure 3-1 Transport Functions of Blood

COMPOSITION OF BLOOD

Because of the significant part which blood plays inmaintaining a suitable environment for other tissue in thebody essentially free of foreign matter and infectious agents,it is inconceivable that changes in the blood would not effectthese other tissues. In recent times we have therefore come tounderstand that, in addition to quantity, the quality of bloodis also significant. In order to understand how blood qualitycan vary, it is necessary to know something about thecomposition of blood.

The blood, which fills the vascular pathways of thecirculatory system, is a truly unique substance. Evenprimitive man probably suspected the vital role that bloodplays and recognized that survival from the hazards of thehunt and combat were in direct proportion to any loss ofblood. We have already indicated that blood performs manyvital functions which suggests that blood is more than asimple fluid. Indeed if blood is examined even casually it canbe seen to be composed of two phases, a fluid phase and acellular phase. This can be easily demonstrated; for example,

Cell Types

Blood Plasma

if a fresh sample of blood is rapidly cooled and centrifugedfor, say, five minutes, it can be separated into a collection ofheavy particles in the bottom of the sample tube leaving aclear fluid on top (supernatant). This clear fluid (about 55%of the sample volume) can be poured off leaving theseparated sediment in the bottom half of the tube. Thesupernatant fluid will be noted after a short while to exhibita unique property, that is, it will congeal into a jelly likemass. This liquid phase of the blood which is capable ofcongealing is called plasma. If fresh plasma is continuouslystirred with a glass rod, it will be found after awhile thatcongealing will not occur. Instead, it will be noted that a paleyellow material (fibrin) will collect on the stirring rod. Theremaining fluid in the tube, which will now not congeal, iscalled serum.

tteturning to the red sediment that was collected, a carefulmicroscopic examination will reveal that a variety of formedelements (cells) have been collected.

Many studies of these cells have been performed and it isknown that the cells in the blood include a least threefamilies of cell types as shown in the tabulation.

A. ERYTHROCYTESB. LEUCOCYTES

1) Granular Polymorphonuclear Leucocytes(Granulocytes)a) Neutrophilsb) Eosinophilsc) Basophils

2) Lymphocytes3) Monocytes

C, PLATELETS

Hence, it seems that whole blood consists of at least fourfunctional elements: plasma, erythrocytes, leucocytes, andplatelets.

The plasma has been demonstrated above to include severalcomponents, since fibrin could be isolated by stirring.Actually, plasma is a complex solution of electrolytes andproteins. The concentration of these plasma constituentsas found in normal blood is tabulated.

ELECTROLYTE COMPOSITION IN BLOOD PLASMA (m eq/liter)

Na+ 140 HCO-3 27

K+ 4 CI. 103

Ca++

mg++

4

2

Proteins-

Misc

15

5

150 150

Fibrina whitish insolubleprotein that forms the essentialportiOn of the blood clot.

19

,Function ofPlasma

20

For the specific roles of each component of plasma, thereader is referred to the abundant literature on hematology.Some of the generalizations which we can note here ascribespecific roles for plasma in (1) neutralizing the acid endproducts of metabolism, (2) regulation of heat loss from thebody, (3) regulation of the proportions of water in thevascular and interstitial compartments, (4) prevention ofexcessive hemorrhage from wounds, and (5) immunologicaldefense against invading viruses and foreign protein.

The formed cells identified previously are also specializedwith respect to their function in the overall role of blood.The erythrocytes, responsible for the transport of gases, arespecialized with respect to form and also chemically by thepresence of the protein hemoglobin. The special shape oferythrocytes, a sort of flattened sphere with a centraldepression (Figure 3-2) results in a cell having a large surfacearea to volume ratio, which improves the efficiency withwhich gases diffuse across the.cell membrane. Simultaneouslythis innovative form achieves a low viscosity for efficientpumping by the heart. The presence of hemoglobin inerythrocytes is principally responsible for the gas transporteffici6cy of blood. Analysis indicates that at sea levelpressures the oxygen dissolved in blood would beapproximately 3 ml per liter of plasma. Under similarconditions the presence of hemoglobin increases the oxygenconcentration to approximately 200 ml. The actual mannerby which gases are taken up by the red cells is not nearly assimple as this discussion may imply and the reader is referredto the many good texts for an in-depth treatment of thissubject.

Nominally7.2 microns

Figure 3-2 Erythrocyte

2.2 microns

The leucocytes, as noted previously, comprise a spectrum ofcell types. Observations made with blood suggest that thesecells have been specialized to remove bacteria and foreignparticles from the blood. Other evidence suggests that thebulk of the activity of leucocytes, however, takes placeoutside of the vascular pathways. The leucocytes therefore

Hemoglobintheoxygen-carrying pigment of theerythrocytes

Plasmathe fluid portion of theblood in which the corpusclesare suspended.

EnvironmentalConditions

are transient residents of the blood stream on their way tosome location in the tissue where they may be needed todestroy invading bacteria or particulate matter. Leucocytesperform their function by a process of phagocytosis(ingestion and digestion).

No one knows with any certainty the origins of each of thecell types nor is the exact role of each in the body's defenseknown. Some generalizations that can be made, however, arethat the three varieties of granulocytes are transported by theblood to tissue sites where they disintegrate releasing activeenzymes, (1) the neutrophils that are specifically active inphagocytosis of bacteria, (2) the lymphocytes that circulatecontinuously between the tissue and bloodstream and areactive in immune responses, and (3) the monocytes that arecirculating phagocytes active in phagocytizing nonlivingparticulate matter.

The platelets are very small and can be easily overlooked inthe blood. They are known to be cell fragments originating inbone marrow and tend to disintegrate readily. Platelets carrysubstantial amounts of serotonin, a vasoconstrictor which isactive in reducing blood loss from severed vessels.Additionally, platelets are believed to play a significant rolein the maintenance of the endothelial lining of the vascularpathways.

ENVIRONMENTAL INFLUENCES ON BLOODPHYSIOLOGY

Most of us would have little difficulty accepting the premisethat disease can influence the form and function of blood.Most are even vaguely aware that the external environment,particularly radiation in the form of x-rays, can causedysfunction of blood. Many, however, will not recognize thatenvironmental changes considered to be physically tolerablemay stress the circulatory system, including thehematopoietic (related to blood forming) process and theimmunological mechanism, beyond acceptable physiologicallimits.

A commonly recognized physiological response to changedenvironmental conditions is found in the deeper and morerapid breathing which is experienced when we go tounaccustomed high altitudes. This accommodative reflex isthe body's response to the hypoxic (low oxygen) atmosphereof the new environment. If this stress persists for a longenough period of time, compensating changes will be made inthe functional elements of the blood; that is, the body willgenerate more erythrocytes (red blood cells) so as to increasethe oxygen-carrying capacity of the blood. This latter changeis an acclimatizing change which minimizes the energyexpended in maintaining an adequate level of oxygen in the

Phagocyte any cell that ingestsmicroorganisms or other cellsand foreign particles.

21

BiochemicalFunctionsInvestigations

22

blood. This acclimating process, however, is not withoutpenalty since the resulting "higher hematocrit" (increased redcell content of the blood) increases the viscosity of the bloodand imposes added pumping loads on the heart. If largeenvironmental changes of this type were permanentlyendured, serious consequences might result. It is to beexpected that even more subtle changes in the blood may beprovoked by other environmental changes. Since some ofthese changes could produce irreversible damage to the body,it is essential to understand how these changes may occur andwhat physiological limits may be endured without harm.

Orbital flight in a spacecraft such as Skylab provides alaboratory having an artificially maintained atmosphere, nearweightlessness, and nearly closed ecological environment.Studies conducted here may provide new informationrelating to man's ability to acclimate safely to spaceflight andmay also provide valuable insight into the underlyingmechanisms by which these changes are accommodated.

The Skylab Hematology and Immunology experimentsdescribed on the following pages include five investigations toevaluate specific aspects of man's immunologic andhematologic systems that might be altered by or respond tothe spaceflight environment. The biochemical functionsinvestigated with these experiments include (1) cytogeneticdamage to blood cells, (2) immune resistance to disease, (3)regulation of plasma and red cell volumes, (4) metabolicprocesses of the red blood cell, and (5) physical-chemicalaspects of red blood cell functions.

The investigations being conducted are not meant to providean all inclusive coverage of immunohematologic functions,but are expected to serve as sensitive indicators ofstress-induced changes in these functions in man. Selection ofthese specific protocols has been based upon experiencegained from previous manned spaceflight missions andassociated ground based studies. The specific goals of theseexperiments are to determine-

1) environmental factors r-::;ponsible for the loss of red cellmass noted during the Gemini Program;

2) relative roles of the 100% oxygen environment, nitrogenin small amounts, weightlessness, duration of exposure,diminished red cell production versus increaseddestruction, ambient total pressure, vibration, ambienttemperature, and dietary factors;

3) spaceflight factors responsible for changes in plasmavolume;

Hematocrit the volumepercentage of erythrocytes inwhole blood

CytogeneticStudies ofthe Blood

4) influence of long-duration spaceflight on the coagulationprocess, platelet function, and vascular friability, and toassess the environmental factors responsible for suchchanges;

5) any alterations in inflammatory response that may occuras a result of long-duration spaceflight, and evaluate;

6) extent to which spaceflight may influence mitosis (celldivision) and/or chromosomal composition, and whichenvironmental factors are responsible and whatpreventive action can be taken if such changes do occur.

SKYLAB EXPERIMENT Mill

EXPERIMENT BACKGROUND

Because chromosome aberration yields in peripheral bloodleucocytes have been found to be sensitive indicators ofexposure to radiation, such measurements can be employedas in vivo dosimeters.

In mitosis (cell reproduction), each chromosome duplicatesitself with the duplicates being separated from each other atcell division. One duplicate chromosome goes into thenucleus of one daughter cell and the other duplicate goes intothe nucleus of the second daughter cell. The end product ofthis process is cell division which involves several phases.Each phase is characterized by a particular pattern ofchromosome behavior. It is during one of these phases (themetaphase) that chromosomal aberrations may bemicroscopically observed.

Chromosome analyses were done for all of the Geminimissions (with the exception of Gemini VIII which wasterminated early) under the operational medical program.Significant, though slight, increases in some types ofchromosomal aberrations were seen following some of themissions. This effect could not be correlated with missionduration, extravehicular activities, isotope injection of thecrews or other obvious flight parameters. Observations on theSkylab crewmembers can assist in elucidating the mechanismof this phenomenon.

Measurement of the number of chromosome aberrations hasbeen demonstrated by ground based studies to be a sensitivemethod of biological radiation dose estimation. Ambientradiation encountered during long duration missions orunexpected solar flare events could produce significantincreases in aberration levels. Even if no detectable increasesin aberration levels are observed in the Skylab missions, theexperiment will have served the useful purpose ofdemonstrating the lack of a detectable genetic hazardassociated with these missions.

Metaphasethe middle stage ofmitosis during which thelengthwise separation of thechromosomes in the equatorialplate occurs.

23

StandardStatisticalProcedures

24

The method of detecting chromosomal aberration will bevisual analysis that involves counting the chromosomes, thenumber of breaks, and types where possible, and then makinga comparison between the identifiable chromosome formswith groups of chromosomes that compose the normalhuman complement.

Standard statistical procedures will be used to determine if asignificant increase in aberrations appears postflight. Thisanalysis will include comparisons of preflight aberrationlevels in normal individuals of the general population."Predicted" aberration levels for postflight samples will becalculated by using inflight physical dose radiationmeasurements (available from Experiment D008, Radiationin Spacecraft, and other sources) and existing experimentallydetermined chromosomal aberration production coefficients.The effects of any other operational or experimentalprocedure likely to produce chromosomal aberration (such asradioisotope injections) will be measured on normal controlsubjects. These control subjects will be similar in age andphysical attributes to the crewmembers. The control subjectswill participate in all tests and medical procedures that areundertaken by the flight crewmembers. Examination will bemade of the chromosomes of the control members and theflight crew before initiation of preflight procedures and teststo detect any chromosomal aberrations already present.

By allowing for predicted aberration yields and the yields dueto experimental or operational procedures, any aberrationfrequency difference evident from comparisons of preflightand postflight samples can be ascribable to radiation or otherspace parameters.


The objectives of this experiment are to make preflight andpostflight determinations of chromosome aberrationfrequencies in the peripheral leucocytes of the Skylab flightcrewmembers and to provide in vivo dosimetry. These datawill add to the findings of other Skylab cytologic andmetabolic experiments to determine the geneticconsequences of long-duration space travel on man.

EQUIPMENT

No inflight hardware is required since the experiment usesblood samples taken pre- and postflight beginning one monthbefore launch and ending three weeks after recovery.

PERFORMANCE

The leucocytes will be placed in a short-term tissue culture.During the first cycle of mitotic activity in the in nitrocultures, standard chromosome preparations of theleucocytes will be prepared.

Dosimetrythe accurate andsystematic determination ofdoses.

Man'sImmunity,in vitroAspects

The leucocytes from the cell culture will be removed duringmetaphase and "fixed." A visual analysis will be performedwhich involves counting the chromosomes, the number ofbreaks and types where possible, and then making acomparison between the identifiable chromosome forms withgroups of chromosomes comprising the normal humancomplement.

D.A.TA

The data from this experiment will consist of thechromosome aberration frequencies that appear after flightfor nine men, three of whom will have experienced 28 daysin Earth orbit and the rest 56 days each. An estimate of theradiation dose experienced by each man will be made basedon the number of chromosome breaks.



Information on man's humoral and cellular immunity andcoagulation phenomena during and following exposure tospace flight is essential before flight crews can be committedto extended missions. Significant alterations of the immunitymechanisms will produce prejudicial effects upon thisinherent defense system, thereby seriously compromising thecrewman's operational status. The cellular immunity systemis exquisitely sensitive to radiation, and the coagulationstatus is affected by man's activity.

The experimental program measures items which contributeto man's ability to combat infections and repair traumatized(injured) tissues after exposure to weightlessness, spacecraftatmosphere, sublethal ionizing radiation, the monotonousimmunologic stimulation of a closed environment, and theunusual orientation and physical activity. Significantalterations of the extracellular or cellularimmune-mechanisms may produce detrimental effects uponnormal physiological functions, may result in increasedsusceptibility to infections, and conceivably can induce theonset of autoimmune diseases.


The objective of this experiment is to assay changes inhumoral and cellular immunity as reflected by theconcentrations of plasma and blood cell proteins, blastoidtransformations, and the synthesis of ribonucleic acid (RNA),and deoxyribonucleic acids (DNA) by the lymphocytes(white cells in lymph).

Auto immunizationtheproduction in an organism ofreactivity to its own tissues

25

EQUIPMENT

The inflight blood collection system will draw venous bloodand centrifuge the samples for preservation. Onboard freezingwill be used to preserve the samples during the mission andmaintain them in a frozen state. They will be returned in thefrozen state in a urine-return container for postflight analysis.This inflight blood collection system and allied facilities willbe used to obtain, process, preserve, and return hematologysamples for Experiments M071, M073, M112, M113, M114,and M115. (See M115 for more details on this equipment.)

PERFORMANCE

Experiment The experiment will obtain preflight baselines, which will beInvestigations indications of normal metabolism, from the crewmembers

and a control group composed of three men physicallysimilar to the crewmembers who will serve as controls whilethe crewmembers are in spaceflight. Inflight blood sampleswill be taken four times from each crewman during the28-day mission and eight times from each crewman duringthe 56-day missions. Upon recovery after the spaceflight,information will be again obtained from the crewmembersbefore body functions "normalize," and compared with thepreflight baselines, inflight profiles, and with the data beingobtained from the ground control group (GCG) to detect anysignificant deviations. An extensive battery of analyses willbe performed using appropriate laboratory techniques todetect qualitative and/or quantitative changes. Periodicexaminations will be made of the blood proteins andlymphocytes until the possible altered concentrations arelikely to have stabilized. Protocols for the experiment aregiven in the section on Experiment M115.

DATA

Data will be gathered from blood samples taken preflight,inflight and postflight. These samples will be analyzed for thefollowing constituents:

1) total plasma proteins;

2) plasma protein fractions (albumins, alpha globulins, betaglobulins, gamma globulins);

3) lymphocyte morphology;

4) ribonucleic acid (RNA) and deoxyribonucleic acid (DNA)synthesis rates in lymphocytes;

5) kinetics of lymphocyte RNA and DNA;

6) observation of blastoid formation;

7) lymphocyte functional response to antigen;

8) quantification of 4.5s, 7s, and 19s.26

Antigena substance thatstimulates antibodies

BloodVolume andRed Cell LifeSpan



Environmental factors encountered during spaceflight mayaffect the red cell mass and plasma volume. These includenear weightlessness, the spacecraft breathing atmosphere, andionizing radiation environments.

Weightlessness may cause a redistribution of the total bloodvolume between the arterial and venous segments and byreducing the quantity of blood in the limbs. In the weightlessstate, a redistribution of pulmonary blood volume may occurwith relatively greater perfusion in the apex of the lungs. Inthis condition, ventilation and perfusion ratios might beexpected to be similar to those produced when the subject islying down in zero gravity. Results from previous missionssuggest that these changes have not produced pathologicalchanges in the blood vessels. It is not known, however,whether long-duration missions could predispose to changesincluding thrombosis (blood clot). Previous missions havealso led to changes in circulating plasma volume at recovery.Analysis has suggested that these changes are of a magnitudesimilar to those found in bed rest. The rate at which thischange occurs and whether the plasma volume decreases or isaugmented during the stress of reentry is unknown. It isbelieved that plasma volume changes may have contributedto the temporary orthostatic hypotension and to the decreasein exercise tolerance that crewmembers experienced soonafter recovery.

The site of red blood cell (RBC) production in the matureadult is the marrow of membranous bones (e.g., sternum andvertebrae). The rate of production is dependent on metabolicdemands and the current red cell population. The rate ofRBC production can be measured quantitatively by injectionof a known quantity of a radioactive iron tracer. Theradioiron, combined with globulin, is transported to otherparts of the body. That part of the iron which reaches themembranous bones is incorporated into the heme portion ofhemoglobin by the bone marrow. Since not all the ironappearing in the plasma is used for erythrocyte productionbut is instead taken up by the iron pools of the body, afraction of the injected radioiron will be unavailable forincorporation into developing RBCs. This can be determinedby measuring the concentration of radioiron in thecirculating RBC after seven days and comparing it with theinitial concentration of radioiron in the plasma.

Since the rate of RBC production acts with RBC loss toincrease or decrease the total RBC mass present at a giventime, any changes in the rates of RBC production anddestruction will be necessarily reflected in the red cell mass.

Hemethe nonprotein,insoluble, iron protoporphyrinconstituent of hemoglobin

27

Red Cell LifeSpan

28

Such changes in red cell mass can be measured and analyzedby injection of radioactive chromium (in the form of sodiumchromate) tagged red cells. The sodium chromate diffusesthrough the cell membrane where it is converted tochromium chloride and, in this form, bound to hemoglobin.The volume of RBCs is then calculated by allowing thechromium-tagged cells to disperse through the circulatorysystem and measuring the extent to which the chromium hasbecome diluted. The fact that chromium does not reenter thered cell makes it a good tracer for RBC mass determinations.Chromium incorporated into the hemoglobin structure of thecirculating red cell also provides a means for estimating therate of random cell destruction by monitoring the rate atwhich chromium disappears from the red cell mass.

To determine selective age dependent erythrocytedestruction and mean red cell life span, carbon 14 labeledglycine will be injected into a superficial arm vein of eachcrewmember and control subject. The radioactively labeledglycine, by its incorporation into the heme portion ofhemoglobin, labels the erythrocytes produced on that day.Sequential blood sampling will then give the percentage oflabeled erythrocytes in the blood at any given time (days),and by plotting this data, survival and destruction curves canbe obtained. The resultant curves can then be analyzedmathematically and a mean life span for the cells determined.Since only a small portion of the carbon 14 is reutilized inerythropoiesis, it is an ideal RBC label.

Finally, plasma volume changes can be measured by adding aknown amount of radioiodinated human serum albumin toeach crewmember's blood. Albumin is a major constituent ofthe plasma and is the protein most responsible formaintaining the osmotic pressure at the capillary membrane.It acts to prevent plasma fluid from leaking out of thecapillaries into interstitial space.

Erythropoiesisthe productionof erythrocytes

The Skylab life support system provides a combination ofbreathing gases which because of the lower than normalnitrogen partial pressure will be hypobaric. The partial Hypobariccharacterized by lesspressure of oxygen in the breathing atmosphere will be than normal pressure or weightslightly increased at least during extravehicular activity bythe crew. Additionally, the atmosphere will contain traceamounts of other gases released from onboard equipment andsupplies. Small amounts of other gases may also be releasedas a result of the crew's metabolic activity and from bacteriaand molds which may be present.

In the Gemini flight, red cell mass decreases were found thatwere generally greater than the red cell mass decreases foundin Apollo flight crewmembers. Total pressure of theatmospheres in the Gemini and Apollo missions were quitesimilar while partial pressures of oxygen and nitrogen were

RedBlood Cell -Metabolism

differentmore oxygen and no nitrogen present in theGemini missions. It is probable that there were alsodifferences in the trace gases respired during the missions.

The red cell mass differences in these previous missions wereprobably related to the above factors although other factorsmight have been important also. As an example, red cell masschanges are expected as a result of changes in the amount ofphysical activity performed during the mission. Athleticconditioning is associated with an increase in red cell mass.Conversely, red cell mass declines have been found in certainbed rest studies, although in the studies to date the amountof blood drawn for research testing may have been a factor inthe decreased red cell mass.

Since a number of inflight environmental conditions havebeen found capable of affecting the total blood volume andits constituents, delineation of inflight effects unhematopoiesis is necessary in order to predict the course and Hematopoiesisthe formation ofconsequences of long duration spaceflights on future flight blood cellscrewmembers.


The objective of this experiment is to determine the effect ofEarth orbital missions on plasma volume and red blood cellpopulations with particular attention paid to changes in redcell mass, red cell destruction rate, red cell life span, and redcell production rate.

PERFORMANCE

Blood samples will be collected preflight, postflight, andinflight, and processed in accordance with the protocOlshown in M115.

DATA

The data to be collected in support of this experimentinclude the following variables:

1) red cell life span;

2) red cell production, distribution, and destruction rates;

3) plasma volume.



At one time, the red blood cell was believed to be an inertparticle composed of water and hemoglobin. We now know,

29

Influence ofSpacecraftEnvironment

30

of course, that the erythrocytes are living cells, doing work,and requiring energy just as other tissues in the body do. Theaverage life span of the human red blood cell is estimated tobe 120 days. During this period, it is estimated that theaverage erythrocyte travels 100 miles between the heart andthe tissue that it serves. In order to remain functional, the redcorpuscle must (1) maintain an internal environment againsta steep ionic gradient across the cell membrane, (2) resistforces that try to change its characteristic bi-concave shape tospherical, (3) maintain active transport mechanisms thatsupport the metabolic requirements of the cell. Interruptionto these processes would render the red blood cell ineffectualand would be fatal in a matter of days.

This experiment will assess the influence of the spaceflightenvironment on the metabolic processes of red cells. The redblood cell requires for its energy source a continuous supplyof glucose.. The process by which the glucose penetrates thered blood cell's membrane is not known; however, it isbelieved to involve an active transport process rather thansimple. diffusion. It is suspected that the membrane'sframework, in particular the fatty (lipid) fraction of theframework, is functional in this process. Because themembrane is the dynamic component in the active transportprocess, its chemical composition and structural integrity willbe examined by this experiment.

Through the metabolic breakdown of glucose, particularlyADP (adenosine diphosphate) to ATP (adenosinetriphosphate) energy is stored in chemical bonds. Changes inthe glucose metabolic pathway, which may occur as a resultof spaceflight, will be analyzed by examination of several keyintracellular enzymes found in preflight and postflight bloodsamples.


The objective of this experiment is to determine if anymetabolic and/or Membrane changes occur in the human redblood cell as a result of exposure to the spaceflightenvironment.

EQUIPMENT (See M115.)

PERFORMANCE (See M115.)

DATA

The data that will be taken in support of this experiment willbe the preflight, inflight, and postflight values of thefollowing variables:

1) methemoglobin;

Enzymeorganic substancecapable of presenting chemicalchanges in organic substances bycatalytic action, such as indigestion

2) reduced gluthathione;

3) glyceraldehyde-6-phosphate dehydrogenase;

4) pyruvate kinase;

'6) glyceraldehyde-3-phosphate dehydrogenase;

6) phosphofructokinase;

7) hexokinase;

8) phosphoglyceric acid kinase;

9) acetylcholinesterase;

10) lipid peroxide levels;

11) adenosinetriphosphate;

12) 2,3-diphosphoglycerate.

Special SKYLAB EXPERIMENT M115HematologicEffect EXPERIMENT BACKGROUND

Data collected from pre-Skylab spaceflights indicate that thespaceflight environment may induce a loss of red blood cellsin the crew. These earlier investigations have suggested thatthis loss may result from a self-limiting hemolysis which Hemolysisthe liberation ofappears to be related to alterations in mean corpuscular hemoglobin

volume and increased osmotic fragility. Since reticulocyte(an early form of the red blood cell) counts which indicatebone marrow activity have not revealed depression of thebone marrow activity due to such flights, and sincereticulocytosis (above normal reticulocyte count) did notappear until the fourth day, it is likely that red cell lossresulted from a reduced need for red cells for the transport ofoxygen.

Similar data gathered during Apollo missions provided a basisfor comparison with the data derived from the Geminiprogram. Comparison of these data suggested that a causativefactor in the reduced RBC was the Gemini atmosphericcomposition7specifically, the high oxygen concentration.Data obtained from the later Apollo flights have alsoindicated alteration of fluid and electrolyte balance, whileexamination of red cell electrolytes by electron probeanalysis indicates similar shifts.

A "bestfit" hypothesis based on these previous data wouldhave it that the hyperoxic atmosphere in spacecraft (andpossibly other environmental parameters such as

31

In flight BloodCollection

32

weightlessness) induces chemical changes in the red blood cellmembrane. These changes directly or indirectly disrupt theactive transport mechanisms leading to increased osmoticpressures and eventual lysis (destruction) of the cell eitherthrough membrane swelling and rupture or through lipidbreakdown and fragmentation.


The objectives of this experiment are to examine criticalphysiochemical hematological parameters relative to themaintenance of homeostasis, and to evaluate the effects ofspaceflight on these parameters.

EQUIPMENT

The inflight blood collection system that supports thisrelated series of studies is designed to collect and processblood samples without coagulation or contamination. Theinflight blood collection system contains the following majorcomponents:

1) syringes capable of extracting approximately 11milliliters of venous blood from each crewman under Venousrelating to a vein.sterile procedures;

2) small blood sampling vials, into which the crewmanplaces a few drops of whole blood already containing afixative;

3) automatic sample processors, two-chamber spring-loadedcontainers, used to process and separate the blood cellsfrom the plasma by centrifugation;

4) a two-speed centrifuge used to receive the automaticsample processors and separate the blood into red cellsand plasma, and to maintain this separation in theweightless environment.

PERFORMANCE

The blood samples for the group of Hematology andImmunology Experiments will be collected in accordancewith Figure 3-3. Specific handling and processing procedureswill be followed to obtain the data for each of theexperiments.

Records will be maintained that include sampling times andthe time, name, and quantity of any injections taken(including x-rays), plus any illness of a subject. A record willbe maintained during the flight of the following events: flightduration, extravehicular activity, solar events, any abnormalexposure to radiation, infectious diseases, symptoms of

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illness, use of medication and types of drugs used, withdosage and duration of usage. Also any deviation from theexpected orbital workshop atmospheric total pressure orpartial pressure of its constituents will be recorded.

Blood samples will be taken inflight four times from eachcrewman during the 28-day mission and eight times fromeach crewman during the 56-day mission.

DATA

Data in support of Experiment M115 will be taken preflightand postflight only and will consist of information about thefollowing variables of routine hematology:

1) red cell count;

2) hematocrit;

3) hemoglobin;

4) red cell indexes;

5) reticulocyte count;

6) white cell count;

7) differential and morphology;

8) platelet count;

9) acid and osmotic fragilities.

and special hematology:

1) red cell critical volume and red cell volume distribution;

2) hemoglobin characterization;

3) single cell hemoglobin distribution;

4) membrane and cellular ultrastructure;

5) intraerythrocytic electrolytes;

6) red cell electrophoretic mobility;

7) red cell density separation;

8) cellular RNA, protein distribution.

Section 4Cardiovascular Status

Lower Body Negative Pressure,Skylab Experiment M092

Vectorcardiogram,Skylab Experiment M093

Blood Flow

GENERAL BACKGROUND

The cardiovascular system, one of the vital systems of thebody, functions as a transport media linking man's internalenvironment (the interstitial fluid surrounding all cells) withthe three large surface areas exposed to his externalenvironment (lungs, gastrointestinal tract and kidneys). Inessence, it consists of a very large number of very smalltubular vessels (capillaries) which perMeate all regions of theinterstitial fluid and the externally exposed areas oflungs, etc. The extensive capillaries are joined by largervessels (arteries and veins) to a pump (the heart), thusforming a closed system through which the transporting fluid(the blood) is kept flowing at an adequate rate.

The circular nature of the blood flow is demonstrated in theschematic diagram of the cardiovascular system of Figure 4-1.The system consists of two .-apillary networks connected inseries with two pumps to form a closed loop. The twopumps, the right and left ventricles of the heart, shownphysically separated in the diagram, actually comprise asingle organ. The blood leaving the right ventricle flows tothe capillary bed of the lungs while that leaving the leftventricle flows to capillaries distributed throughout the restof the body. Each ventricle is preceded by a supportingatrium, a chamber of the heart, which acts as a receivingreservoir and auxiliary pump to fill the associated ventricle.

Blood leaves the right ventricle through a single, large vessel,the pulmonary artery, and enters the left atrium from the

Pulmonaryartery(15 mm Hg)

Rightventricle

PulmonaryiAcapillaries146(.6 mm Hg)

Rightatrium(0 mm Hg)

Venoussystem

,---Pulmonaryvein

Leftatrium(6 mm Hg)

Average mean pressures areindicated in parentheses

Flow

0111111111or

SystemicgqscapillariesN(25 mm Hg)

Leftventricle

Arterialsystem(90 mm Hg)

Figure 4-1 Schematic Diagram of the Circulatory System

Arteriesoxygenated bloodVeinsunoxygenated bloodcarrying carbon dioxide

35

SystemicCircuits

lungs by way of four pulmonary veins; it leaves the leftventricle through the large aorta, and returns from all parts ofthe body by way of two large veins, the superior and inferiorvenae cavae. All vessels carrying blood from the heart tocapillaries are arteries, and those conveying blood fromcapillaries back to the heart are veins. The circulating bloodvolume in man is about 76 ml/kg of body weight, about 5.3liters in an average man weighing 70 kilograms.

The part of the vascular system traversed by the blood ingoing from the left ventricle to the right atrium is called thesystemic circuit. It includes the capillary beds of all parts ofthe body except the lungs. The pumping force that movesblood through this circuit comes from contractions of theleft ventricle. The large number of capillary networks thatsupply the organs and tissue of the body constitute manycircuits connected in parallel and supplied by the extensivelybranched arterial system that begins with the aorta. Bloodleaving the capillaries enters small veins that join together toform larger and larger veins. This arrangement continues untilall the systemic blood returns to the right atrium through thetwo large venae cavae. The systemic circuit contains about80% of the total circulating blood volume, approximately4240 milliliters for the 70-kilogram average man.

Capillaries The capillaries, which have very thin and permeable walls,exchange water and dissolved materials (including gases) withthe interstitial fluid. This two-way passage (in and out) offluid has been estimated to exceed 200 liters per minute.

The capillaries are very small but their number is so large thatthe total area available for diffusion is very large, approaching100 square meters. The number of capillaries in tissue can becorrelated with the metabolism of the tissue; for example, ina heart muscle where oxygen use is high, microscopic studiesshow as many as 6000 capillaries in each square millimeter oftissue cross section. Since many capillaries may "closedown," especially in the skeletal muscle except duringexercise or work, the volume of blood present in the systemiccapillaries is variable. It has been estimated that in an averageadult this volume may range between 60 and 200 milliliters.

The systemic arteries act as conduits to distribute blood tothe systemic capillary beds. The aorta at its beginning (theascending aorta) has an outer diameter of about 2.5centimeters and a wall thickness of nearly 2 millimeters. Ateach branch point, the cross sectional area of each branch issmaller than the parent vessel while the total cross sectionalarea of the branches is greater. The arteries do not contributesignificantly by diffusion to the exchange of materials withthe interstial fluid. The volume of blood contained within allthe systemic arteries under normal circumstances is estimatedto be about 20% (one liter) of the total blood volume.

36

Total Blood Volume"'5 litersVolume in SystemicArteries"-1 liter

PulmonaryCircuit

The smallest branches of the arterial system are calledarterioles. Arterioles are different from small arteries in thattheir walls are thick in relation to their internal diameter.This extra wall thickness consists of circumferentiallyoriented smooth muscle fibers. The cross sectional area of thearteriolar lumen varies with contractions of the smoothmuscle which are controlled by neurohumoral factors. Thesechanges control resistance to blood flow in the systemiccircuit and distribution of the blood flow to the capillarybeds.

The veins form an extensive recovery system for conveyingthe blood returning from capillary beds to the. right atrium.Although their walls are much thinner than those of thearteries, they also permit no significant diffusion ofsubstances through them. The distinctive feature of thevenous system is its large volume. The volume of bloodcontained within the veins is about 75% of the volume in thewhole systemic circuit or about 3200 milliliters. Veins, unlikeother vessels, are equipped with one-way valves that permitblood flow only toward the heart.

The pulmonary circuit, which is sometimes called "the lessercirculation," extends from the pulmonary artery as it leavesthe right ventricle to the pulmonary veins as they enter theleft atrium. The pulmonary circuit carries venous blood (i.e.,blood which has come from systemic capillaries and istherefore low in oxygen content and high in carbon dioxidecontent) to the lungs, where it is converted to arterial blood(blood which is freshly oxygenated and has had much of thecarbon dioxide removed). The pulmonary artery is the onlyartery that carries venous blood, and the pulmonary veins arethe only veins that carry arterial blood. The pulmonarycircuit, like the systemic, is composed of arteries, capillaries,and veins, but all vessels are generally shorter and larger indiameter than corresponding systemic vessels. In thepulmonary circuit thick-walled arterioles are absent and thevery small arteries (<100 microns in diameter) show little orno smooth muscle in their walls.

In the human lungs, air is conducted via the trachea, bronchi,and their branches to about 300 million small air sacs oralveoli, which are honeycomb-like structures about 200 to250 microns in diameter. The pulmonary capillaries, whichare 10 to 14 microns in length, and 7 to 9 microns indiameter, extend as a dense network within the thin,membranous partitions separating adjacent alveoli.Calculations have shown the surface area of capillary wallexposed to the alveolar air averages to be from 50 to 70square meters in the average adult. The total volume of bloodwithin the capillaries of the pulmonary circuit is of the orderof 100 milliliters.

Microna millionth part of ameter

37

DYNAMICS OF THE CARDIOVASCULAR SYSTEM

Much of our knowledge of the dynamic characteristics of thecardiovascular system, and the mechanisms governing itsresponse to changes in activity, environment, and disease hascome from observations on animals, chiefly dogs. In recentyears, great technological advances in bioinstrumentationhave made it possible to confirm and extend theseobservations on man. It is still true, however, that manymeasurements can be made only on anesthetized animals,some can be made as well on unanesthetized animals but noton man, and some on both animals and man in the consciousstate. Under all conditions, the method of measurement usedwill disturb, to some extent, the cardiovascular system of thesubject, and influence the variable being measured. It isalways important to minimize this disturbance, to assess itssignificance, and to take it into account, when possible, ininterpreting the results.

Hem o dynamically, the circulatory parameters of mostinterest are pressures and flows in various parts of thecardiovascular system. Because of the rhythmicalcontractions of the heart, the input of blood into bothpulmonary and systemic circuits is intermittent.Consequently, the blood flow in all the systemic arteries, thesystemic veins close to the heart, and probably all of thepulmonary vessels is pulsatile rather than steady. In thesystemic capillaries and peripheral veins, the flow isessentially steady because of the low-pass filtering action ofarterial compliance and arteriolar resistance. Pressures,therefore, are pulsatile it. all four chambers of the heart andin all parts of the vascular system with the exception ofsystemic capillaries and peripheral veins.

Mechanically, the heart is a muscular organ consisting of twomajor pumping chambers, the right and left ventricles, eachof which receives blood from a contractile antechamber, rightand left atria, respectively. Backflow of blood from ventricleto atrium is prevented by the valve on the right side(tricuspid) and the valve on the left side (mitral). The right Rightventricle ejects blood into the pulmonary artery from which Atriumbackflow is prevented by a semilunar pulmonary valve. Right LeftBackflow of blood into the left ventricle from its outflow Ventricle Ventriclevessel, the aorta, is prevented by a similar semilunar aortic Diagram of Humanvalve.

Right LeftLung Lung

LeftAtrium

Heart

Heart An outstanding characteristic of cardiac activity is the fact AutomaticityoccurringAutomaticity that the heart continues to contract rhythmically when all of spontaneously, unvoluntarily

its connections with the nervous system are severed, or evenwhen it is removed completely from the body. This indicatesthat, unlike skeletal muscle, cardiac muscle is not dependentupon impulses coming to it from the nervous system for itsactivation. It has the property of automaticity.

38

ContractionSequence

Problems Dueto Lack ofGravity

Normally, the rhythmically occurring stimuli responsible forcardiac contraction arise in the sinus node. This "built-instimulator" or pacemaker of the heart has the property ofautomatically discharging rhythmical stimuli which activatethe heart. This wave of excitation, beginning in the sinusnode and traveling in all directions through the atrial muscle,is known as the cardiac impulse. It consists of electrical(action currents), chemical and thermal changes, and isfollowed closely by mechanical contraction. As in the case ofa nerve impulse traveling along a nerve fiber, or in impulsetraversing a single skeletal muscle fiber, the cardiac impulse Depolarizationthe act ofalso involves a progressive depolarization of cell membranes. neutralizing polarity

The normal sequence of contraction of the heart chambersresults from the spread of the cardiac impulse. This impulsearises in the sinus node, located in the wall of the rightatrium; hence this chamber contracts first. Because the rateof conduction of the impulse through atrial muscle is quiterapid, the left atrium contracts only a very short time afterthe right. Practically, the two atria contract almostsimultaneously. The impulse is slowed down considerably,however, while traveling between the interatrial septum andthe ventricular muscle mass. This is because conductionbetween atria and ventricles can only occur by way of the

A-V (atrio-ventricular) node and "bundle" (a specializedconducting tissue). The only other tissue normallyconnecting auricles with ventricles is connective tissue, whichdoes not have the property of conducting an impulse. Thisresults in the completion of atrial contraction beforeventricular contraction begins. During the period ofcontraction and depolarization, action currents similar tothose in all muscles of the body can be sensed at the surfaceof the body.

ENVIRONMENTAL INFLUENCE ON CARDIOVASCULARFUNCTION

During routine daily activities, the force of gravity influencesthe distribution of blood in the cardiovascular system and,consequently, changes the nature of blood flow regulation.Thus, the function of this system is dependent upon theeffects exerted by the Earth's gravitational pull.

The effects of null or diminished gravitational forces that areexperienced during space flight have not been fullyelucidated by past studies. However, flight observationscoupled with analyses of the cardiovascular system haveindicated the following potential problem areas when theforces of gravity are altered.

Cardiac ContractilityReduction in work performed byantigravity muscles reduces the nutritive and waste removal

39

requirements imposed upon the cardiovascular system; hence,the heart leads a more sedentary existence. One mechanismby which it can decrease its total output is to reduce theforcefulness of each contraction.

Vascular ResponsivenessAlterations in the responses of thevascular smooth muscle, which occur with decreasedstimulation (hormonal, nervous, mechanical) with changesin the contractile properties of smooth muscle cells, and withalterations in vessel geometry caused by change inintravascular volume, will result in qualitative andquantitative changes in the cardiovascular function. Inparticular, the vascular neural reflexes that compensate fordrainage of intravascular fluid away from the heart when aman assumes an upright position in a gravitational fieldbecome less effective. This situation, called orthostatichypotension, can develop after a few days in a weightlessstate and upon return to Earth could lead to temporary boutsof fainting because of inadequate brain circulation.

Capillary Exchange Alteration in the transcapillary pressuregradients or the properties of the capillary walls change thequantity and the composition of the fluid that moves fromthe intravascular into the extravascular spaces.

Viscoelastic Characteristics of VesselsChanges in vasculardistensibility, especially that of the capacitance vessels(located principally in the venous circulation), markedly alterthe quantity of blood returned to the heart as well as thetime-dependent characteristics of venous blood return, andthence, alter the output of the heart.

Decreased Endocrine ActivityDepletion of vasoactive vasoactiveexerting an effecthormone stores (e.g., norepinepherine) or decreases in their upon the blood vesselsrates of synthesis effectively limit or alter the ability of thevascular system to respond to stress when the need arises.Smooth muscle responses to given concentrations ofvasoactive hormones may also assume different patterns asadaptation to a new environment occurs.

Decreased VascularityBoth an enriched oxygen atmosphereand a decrease in oxygen demands of tissues associated withdecreased work will reduce the oxygen transportrequirements placed upon the cardiovascular system. The rateof oxygen diffusion through the tissue spaces to meet cellularoxygen needs will become lower. Thus, the diffusion pathscan be lengthened without compromising these needs. Underthese circumstances, tissues become less vascular through arelatively slow process of acclimatization.

Shifts of Intravascular Fluid VolumesVolume receptors thatrespond to vessel or heart chamber wall stretch are stimulatedby shifts of fluid within the vascular spaces when

40

Inflight LowerBody NegativePressure(LBNP)

gravitational forces are removed or decreased. Throughneuroendocrine pathways, a diuresis is initiated and the totalwater content of the body is reduced. This results in partialdehydration of body tissues and reduces the capacity torespond to stress.

Nonspace Related ConditionsConsideration must also begiven to debilitating cardiovascular diseases and to otherEarth environmental conditions not unique to weightlessness.The occurrences of these are not precluded by weightlessness;hence, long term comprehensive longitudinal studies on flightcrews and improved prediction models are required toestablish probability tables to assist medical support of longduration missions.

The Skylab program incorporates two experiments thatprovide information on the performance of thecardiovascular system in the near weightless environment ofspace. This information will aid in establishing the relativeimportance of the above problems for long-durationspaceflight.



Reduced orthostatic tolerance can be demonstrated by Orthostaticpertaining to orprovocative (stimulating) testing during weightless flight. One caused by standing erect.method of provocative testing involves the application oflower body negative pressure (subambient pressure to thebody surface below the diaphragm). The equipment to applythis negative pressure is the lower body negative pressuredevice (LBNPD). Physiological indicators are heart electricalactivity, pulse pressure, blood pressure, heart rate and limbvolume.

The body stresses produced by LBNP on the cardiovascularsystem of a supine subject in 1-g in certain respects closely Supinelying on the back in aresembles the effects produced by a normal upright posture.These stresses are an effective decrease in resistance to bloodflow to dependent portions of the body, and increase in thelevel of venous pressure in dependent veins required to returnblood to the heart. LBNP results in pooling of blood in thelower body hindering its return to the heart, and thereforeelicits a similar cardiovascular response to that experiencedduring' upright posture on Earth. LBNP thus provides anobjective means of provoking cardiovascular responses thatcan be used to determine the extent of cardiovasculardeconditioning. The technique could conceivably serve as acountermeasure to deconditioning due to the near weightlessenvironment, by providing periodic calibrated stresses thatmay minimize the effects of spaceflight.

relaxed position

41


Cardiovascular The objective of this experiment is to provide informationconcerning the time course of cardiovascular deconditioningduring flight and to provide inflight data for predicting thedegree of orthostatic intolerance and impairment of physicalcapacity which is expected following Earth return.

Deconditioning

42

EQUIPMENT

The major components of the experiment equipment usedare-

1) lower body negative pressure device (LBNPD),2) leg volume measuring system (LVMS),3) blood pressure measuring system (BPMS),4) body temperature measuring system (BTMS) (M171

experiment equipment),5) vectorcardiograph (VCG) (M093 experiment equipment).

The LBNPD consists of a cylindrical tank with a waist sealthat provides positive isolation of the inner volume and lowerlimbs from the ambient atmosphere. An inner crotch supportprevents the crewman from being pulled into the device bythe reduced internal pressure during operation. This devicecan be evacuated to a controllable pressure of 0-50 mm Hgbelow the ambient cabin pressure.

The LVMS senses the changes in the circumference of the legat the level of the calf muscle by a capacitive type detectorleg band. Data signals from the leg band are then conditionedand displayed on a console panel as percent volume change.

The BPMS includes an inflatable arm cuff and a microphonefor detecting the Korotkoff sounds characterizing blood flowusing the auscultatory method. These pressures can bedisplayed and/or recorded for transmission to the ground.

PERFORMANCE

Each of the crewmen will participate in the LBNP tests atapproximately the same time each day, every third dayduring the mission. A LBNP test will consist of a 5-minuteperiod of rest at 15 minutes of progressively decreasingpressure on the lower limbs until 50 mm Hg (1 psi) belowcabin pressure is reached. This is followed by 5 minutes ofrest. Physiologic measurements are made throughout the testprogram.

A minimum of five measurement sessions are planned foreach flight crew at approximate intervals as follows: selectionof the crew, 60 days, 21 days, 10 days, and 2 days beforelaunch. These data will serve as both the experimentalcontrol and the baseline reference for postflight data.

Korotkoff Methodtodetermine blood pressure

The auscultatorymethodlistening for sounds inthe body to ascertain thecondition of the lungs, heart, etc

Postflight measurements will be made beginning as soon aspossible after recovery, at recovery plus 24 hours, and at each24-hour interval thereafter until normal preflight values areobtained.

DATA

The principal measurements taken during the experiment areas follows:

1) pressure differential (AP across the chamber),2) leg circumference (correlatable with leg volume),3) blood pressure,4) internal temperature (LBNPD chamber temperature),5) cabin temperature (ambient temperature to the subject's

head, shoulders, and upper torso),6) subject's body temperature,7) vectoreardiogram,8) heart rate.

Vectorcardi- SKYLAB EXPERIMENT M093ogram


The excitation process in cardiac muscle is similar to that inskeletal muscle and in nerves. It results in a propagateddepolarization of cell membranes described below and ismade evident by action currents. The electrocardiogram is agraphic record of the action currents from the heart muscle.

These action currents may be recorded by placing pickupelPetrodes at two locations on the surface of the body. Atany instant during the cardiac cycle the heart may beconsidered as a battery with the depolarized region of theheart (through which the impulse has already passed at thatinstant) acting as a negative pole, and the inactive region ofthe heart (through which the impulse has not yet passed)acting as the positive pole. This battery is immersed in anelectrically conducting medium, the body. Consequently,action currents will flow, not only within the heart, butthrough all the tissues of the body, including those at thesurface. Hence, surface electrodes may be used to lead off andrecord these potential differences as an index of cardiacaction currents.

Electrocardio- As the cardiac impulse spreads through the heart during eachgram heart cycle, the spatial and electrical relationships between

active and inactive regions of the heart are constantlychanging, thus causing the surface potentials to change frommoment to moment, both in magnitude and direction. Thesechanging potential differences between lead-off points on thebody surface, when graphically recorded, constitute theelectrocardiogram.

Vectorcardiograma graphicrecord of the magnitude anddirection of the electrical forcesof the heart

43

ElectricalAxis of theHeart

MeasurementPoints

44

The electrical condition of the heart at any instant may berepresented by a vector whose direction indicates theorientation of the negative or depolarized portion of theheart with respect to the positive or inactive region of theheart at that instant. The magnitude of this vector isproportional to the instantaneous potential differencebetween the positive and negative portions of the heart. Thisvector, which undergoes cyclical changes in magnitude anddirection in space during each heart cycle, is known as theelectrical axis of the heart. The instantaneous potentialdifference recorded between any two surface lead-off pointsis proportional to the projection of the electrical axis uponthe line-joining these two points.

Thus, a record taken from two points on the surface of thebody will provide information concerning the cyclicalvariations of the electrical axis of the heart. Moreinformation can be obtained by recording from several pairsof surface points than can be obtained by recording fromonly one pair. Hence, it is customary to record, in turn, fromthree pairs of lead-off points known as the three standardlimb leads. Lead I is taken from the two arms, lead II fromthe right arm and left leg, and lead III from the left arm andleft leg. (See Figure 4-2.)

is

X AxisP

U

Z

Typical ECG-Lead

Standard ECG Limb Leads

TimeSI (left arm-right arm)

Legend:

Positive Negative F FrontLead Terminal Terminal B Back

Left Arm (LA) Right Arm (RA)I RS Right SideII Left Leg Right Arm LS Left SideIII Left Leg Left Arm LC Left Chest

Figure 4-2 Standard ECG Connections

If some portion of the heart is injured as the result ofinadequate blood supply or some disease process, injurycurrents will flow. These injury currents may appear in thestandard limb leads, but sometimes, if the injured area issmall, or if it is located in particular regions of the heart, theinjury current can be recorded only by applying oneelectrode to the surface of the body as close to the heart aspossible, i.e., on the chest. Consequently, it is customary to

Electrocardiograma graphictracing of the electric currentproduced by the contraction ofthe heart muscle

Reading theECG Wave

Vectorcardio-gram

record, in addition to the limb leads, two or more chest leadsin which one electrode is placed on the chest and the otherelectrode on an arm or leg. Additional leads are often takenfrom the chest and sacral regions.

The form of the electrocardiogram recorded from any lead isdetermined by the site of origin of the cardiac impulse, itspathway of spread through the heart, the position of theheart within the chest, and the presence or absence of injurycurrents. This is simply another way of saying that the formof the electrocardiogram is determined by the cyclicalvariations in direction and amplitude of the elt,dirical axis ofthe heart and the point of attachment of the leads.

The form of the electrocardiogram for a typical normal heartcycle is indicated in Figure 4-2. This curve is typical for thelimb leads; minor variations are seen in the chest leads. Theimportant characteristics of the curve are the P wave, theQRS complex, the S-T segment, and the T wave. Importanttime intervals are the P-R interval and the duration of theQRS complex.

The P wave results from the activation of the auricles. TheQRS complex results from the activation or depolarization ofthe ventricles. The S-T segment occurs during the time inwhich the ventricles are in the completely depolarized state,and _the T wave represents ventricular repolarization. Nosignificance is attached to the U wave which is sometimesseen.

A specialized form of the electrocardiogram calledvectorcardiogram (VCG) can be obtained by plotting onechannel against the other. Figure 4-3 shows a typicalarrangement for generation of VCGs. A cardiologist trainedin interpretation of VCGs can tell much about the activitiesof the heart from such a display.

Horizontal

Leg

Figure 4-3 VCG Connections

Oscilloscope

Typical VCG (leaps I & V1)

Cardiac Impulsethe impulse orbeat of the heart at the fifthintercostal space at the left sideof the sternum

45

Actually experiment M093 will analyze the three channels ofinformation on a computer to get the information needed.Among the things that can be determined by an analysis ofVCG are the electrical orientation of the heart and itsmovements. This experiment will analyze the VCG taken oneach of the nine astronauts who fly in Skylab both preflight,inflight, and postflight. VCGs obtained while the astronaut isat rest and at a number of exercise levels on the ergometer Erg ometeran instrument forwill also be analyzed. measuring the force of muscular

contraction


The objective of this experiment is to measure thevectorcardiographic potentials of each astronaut periodicallythroughout the mission so that flight-induced changes inheart function can. be detected and compared with a baselineestablished preflight.

EQUIPMENT

The equipment consists of the various electrodes and wiresfor "picking up" the voltages from the astronaut's body andthe electronics equipment for amplifying the voltages,protecting the astronaut from electrical shock, andconditioning the voltages into the three channels ofinformation.

PERFORMANCE

The experiment is scheduled to be performed every threedays on each astronaut. The subject crew member attachesthe VCG electrodes, rests on the ergometer for five minutes,then pedals for two minutes at a set work rate of 150 watts.He then rests for 10 minutes.

DATA

The data, recorded on tape to be telemetered to the nearestground tracking station, consist of the three standard vector-cardiogram voltage signals and a heart rate channel, along withvoice identification of the conditions of recording.

46

Section 5Energy Expenditure

Metabolic Activity,Skylab Experiment M171

LU

C0U

(f)

EnergyProduction

GENERAL BACKGROUND

The most basic process of living systems is production ofenergy by metabolism. The energy derived from theseprocesses is used for physiological and biochemical functions,permitting the organism to perform work.

When an individual is not moving or working and has notrecently eaten, all of the energy appears as heat. During thiscondition, energy production can be directly measured bydirect calorimetry. However, this covers only a small portionof the time when total energy measurements are required.

Energy production can also be calculated using indirectcalorimetry which measures the oxygen consumed. Sinceoxygen is not stored (except for about one liter asoxyhemoglobin and myoglobin in the blood and tissues), itsconsumption keeps pace with immediate needs and isproportionate to the total energy produced. One problemwith using only oxygen consumption as a measure of energyproduction is the fact that the energy released per mol ofoxygen is dependent upon the food being oxidized. Althoughan estimate of 4.82 k cal/mol oxygen is accurate enough formost purposes, the actual values can be determined from ananalysis of the respiratory quotient (RQ), which requires thatcarbon dioxide production (during steady state conditions)be measured simultaneously with oxygen consumption.

The measurement of energy production through the use ofindirect calorimetry has been accomplished clinically and inthe laboratory using two general methodsclosed circuit andopen circuit. Excellent discussions of these two types ofmeasurement can be found (Consolazio, et al. 1963; Best andTaylor, 1961; Bard, 1961; Bartels, 1963.) In the closedsystem the subject rebreathes from a container of 100%oxygen and a carbon dioxide absorber. The oxygenconsumption is then determined by the decrease in volume ofthe system or by the amount of oxygen added to keep thevolume constant. If carbon dioxide production is desired, thecarbon dioxide absorber must be weighed or analyzed, or beassessed by a system that determines volumes before andafter carbon dioxide absorption. Luft (1958) has reviewedspirometric methods which have been used for closed circuitindirect calorimetry.

The standard open circuit method of determining metabolicrate is ba'sed on minute volumes and gas concentrations.Usually only expired volume (collected in a spirometer -Tissot method; in a bag - Douglas method; or measured witha dry gas meter with a portion of gas being saved for analysis -Kofranyi-Michaelis respirometer) is measured and analyzedfor its oxygen, carbon dioxide, and nitrogen content. Fromthese values it is possible to calculate inspired volume andtherefore V02, Ve02, and RQ.

C a 1 o r imetrymeasuremen ts ofthe amounts of heat absorbed orgiven out

47

Life Support

48

If man is to be qualified for long duration missions, it isimperative that energy metabolism data be collected in flight.This is needed because of logistics and life supportrequirements, as well as the possible physiologicaldegradation of work performance.

Until inflight data can be collected, it is also required thatenergy metabolism and work performance, as well aspulmonary function, be evaluated pre- and postflight. Shoulda physiological degradation of ability to do work be evident,an acceptable limit of this degradation will have to beestablished. These limits will be determined by requiredactivities for the successful completion of a mission,considerations relating to the mechanism behind thedegradation, and whether countermeasures are possible.

The life support systems both in the spacecraft and the suitsfor EVA operations must be adequate to accommodate the EVAextravehicular activityfunctional work capacity (both sustained and transientthermal loads) of the typical astronaut. While the workcapacity in the spacecraft is not likely to exceed that of anaircraft pilot in the geogravitational environment, this cannotbe said of EVA operations, where an emergency may raisethe metabolic level to that experienced during heavy exertionon Earth. Fletcher (1964) has compiled data from severalsources (Figure 5-1), to show metabolic curves for first classathletes and healthy men engaged in exerting activities(running, rowing, cycling, etc) under normal atmosphericconditions. The design criteria for the life support systemsfor EVA operations should be such as to permit the levels ofactivity shown by Fletcher, thus providing for thoseemergencies which call for these higher metabolic levels.

2.1.501.25

4) 1.00

0.75cn

0.50

A 0.25

00.1

Time, hr0.25 1 3 5 10

First class athletes

Healthy men

"MI

1.0 10.0Time, min

100

Metabolic RateBtu Kcalhr hr20,000 5,000

3.0 18,000 4,5004-2 14,000 4,000

2.5 0^14,000 --3,5003

2.0 1'12,000-3,00010,000-- 2,5008,000-2,000

1.0 bag 6,000-1,5004,000-1,000

O2,000-500

0-01,000

Figure 5-1 - Maximum Sustained Work Capacity

MetabolicActivity



Metabolism has been an area of investigation from the firstmanned missions. During the Gemini Program in the mid 60s,a number of the medical investigations were concerned withmetabolism. Many tests were performed to determinewhether the crew would become fatigued doing the types ofactivities that had been planned, and whether the life supportsystem in the spacecraft and in the space suit would be ableto meet the astronauts' requirements. During these earlymissions, it was learned that there were some significantproblems. The astronauts became much more fatigued thanhad been expected, tasks were much harder to perform, andthe possibility arose that under certain conditions the carbondioxide levels in the space suit rose to higher than desirablelevels. Problems with the design of equipment and the workregimens specified for the astronauts were discussed. Theseconcerns and investigations continued into the ApolloProgram, and still many questions remain to be answered.

Of particular interest is the relationship between themetabolic requirements of certain physical tasks in spacecompared to the requirements for the same tasks on Earth. Acommon means of determining these requirements is toascertain the oxygen consumption and carbon dioxideproduction of the body during the performance of thespecified task.

These consumption and production rates cannot be measureddirectly at the cellular level where the demand exists.Measuring intake and output for the body as a whole isconsidered to represent the aggregate demand for all cellularactivity. Consumption and production are usually calledoxygen uptake and carbon dioxide output. The former isdetermined by measuring the amount of oxygen that aperson breathes in over a given period of time and subtractingfrom it the amount of oxygen that he breathes out over thesame period of time. The latter is determined similarly: thevolume of carbon dioxide breathed in over the period issubtracted from the volume of carbon dioxide breathed outover that same period. For certain types of exercise, theoxygen uptake is equal to oxygen consumption and carbondioxide output is equal to carbon dioxide production. Byproperly planning an experiment, the oxygen consumptionand carbon dioxide production can be determined from theoxygen uptake and carbon dioxide output measurements(along with other supportive measurements and information).

By doing such determinations while a subject is performing ameasured amount of physical work, the relationship betweenmetabolic activity and the physical work can be determined.

Metabolismthe sum of all thephysical and chemical processesby which living organizedsubstance is produced andmaintained.

49

Such measurements are commonly done in the laboratorytaking the measurements while the subject is performingwork on a treadmill or an ergometer, each of which is ameans of providing known amounts of physical work for theindividual to do.


The objectives of this experiment are to determine if man'smetabolic effectiveness in doing mechanical work isprogressively altered by exposure to the space environment,and to evaluate the bicycle ergometer as an exerciser for longduration missions.

EQUIPMENT

Measurements Five major pieces of equipment are used for this experiment.

1) Metabolic Analyzer determines the oxygen update forone minute and five minute periods; carbon dioxideoutput for one minute and five minute periods; minutevolume, the total gas expired by the subject during aone-minute period; respiratory exchange ratio, the ratioof carbon dioxide output to oxygen uptake; and vitalcapacity, the measurement of the maximum amount ofgas that a person expires after a full inspiration.

2) Ergometer, a controllable workload pedal device, having aworkload selector and tachometer. The pedals connect tothe workload which offers mechanical resistance set to adesired level. The ergometer measures work rate, theactual work rate which the astronaut performs on theergometer; revolutions per minute, the rate at which theastronaut pedals the ergometer; and total workperformed during a given period.

3) Body Temperature Measuring System electricallymeasures the temperature in the subject's mouth using anoral thermistor.

4) Vectorcardiograph senses three channels of electricalsignals from the heart and permits heart rate to bedetermined,

5) Blood Pressure Measuring System comprises amicrophone and inflatable arm cuff.

PERFORMANCE

The metabolic activity experiment will begin 12 monthsbefore flight. The experiment will be repeated at one-monthintervals from 6 months before launch and also 5 days beforeflight. Inflight the experiment will be repeated five times by

50

Thermistor a special type ofresistance thermometer thatmeasures extremely smallchanges in temperature

each crewman during the 28-day flight, and eight timesduring the 56-day flight. After recovery the exercise capacitytest will be performed on each crewman as soon as possibleafter recovery atid again after 24 hours. If there Pre anysignificant changes in metabolic performance during thecourse of the mission, these changes will be followed untilbaseline levels are reached.

Work profiles for this experiment include (1) the subjectremains relaxed and motionless for 5 minutes, (2) he thenpedals for five minutes with an energy output fixed at 25% ofhis preflight determined maximum capacity; (3) the workrate is changed to 50% of his preflight maximum capacity,and the subject operates the ergometer at this rate withoutinterruption for another five minutes; (6) the work rate ischanged to 75% of the preflight determined maximumcapacity and operated at this rate for five minutes; (7) thesubject stops the ergometer, relaxes, and remains motionlessfor 5 minutes.

DATA

The data return from this experiment will consist of theprofiles that occur during a prescribed exercise regime in zerogravity for the following variables: blood pressure, heart rate,vectorcardiogram, and metabolic rate. The metabolic rate iscomputed from measurements of oxygen consumption andcarbon dioxide production. The data derived from thisexperiment are recorded on the spacecraft tape recorder andtransmitted to ground. Manual data recording is available as abackup mode. Voice comments are also recorded. Motionpicture data will be obtained using a 16mm camera.

51

Section 6Neurophysiology

Human Vestibular Function,Skylab Experiment M131

Sleep Monitoring,Skylab Experiment M133

(.0

0

cna.)

HumanVestibularFunction

GENERAL BACKGROUND

The physiology of the nervous system in man is complex,ranging from the sensing and processing of input informationvia the afferent nervous system, to the higher orderneurological functions having a poorly understoodneurophysiological basis, e.g., language, learning, and statesof consciousness.

Two investigations being conducted on Skylab will evaluateaspects of the central nervous system and the impact ofweightlessness. The first, an experiment involving the humanvestibular apparatus, will investigate the effects ofweightlessness on the crewman's ability to maintainperceptual acuity and orientation in space. This experimentwill identify changes in the normal functioning of thevestibular apparatus after long periods of weightlessness.

The second investigation will use the electroencephalogram(EEG) in an attempt to evaluate the sleep patterns derivedfrom m the brainwaves in the space environment. Thisexperiment should establish whether there are changes in thesleep quality or quantity associated with extended flights.



Body position and movement is perceived principally byspecialized sense organs in the inner ear, collectively termedthe vestibular apparatus or the labyrinthine receptors. Theseorgans are the otoliths and the semicircular canals. Theotoliths are stimulated by linear accelerations (includinggravity) and are the specialized organs for the sensoryperception of head tilt relative to the direction of the localforce field. The semicircular canals sense the magnitude anddirection of angular accelerations. The brain integrates thesesensory data inputs to determine and maintain the body'sposture and orientation.

The importance of the vestibular organs in day to dayactivities becomes apparant if one considers theirneuroanatomical connections to the--

1) reticular system, dealing with alertness and attention,2) eye muscles,3) autonomic nervous system, dealing with regulation of

respiration, heart rate, GI tract motility, etc,4) voluntary and anti-gravity body muscles, and

5) cerebral cortex.

Cerebral Cortexthe convolutedlayer of gray substance thatcovers each cerebal hemisphere

53

Reaction ofAstronauts onPreviousFlights

54

Experimentally produced discrepancy between visual,vestibular, and tactile-kinesthetic spatial perceptions havebeen shown to cause a stressful sensory conflict which,depending on the individual, produces symptoms rangingfrom disorientation to nausea and vomiting.

Varying degrees of discomfort and disorientation have alsobeen noted under conditions of near weightless spaceflight.During the first American orbital flight (MA-6, February 20,1962) Astronaut Glenn in his pilot report noted that he hadexperienced an illusion of tumbling forward after cessation ofthe initial acceleration of his vehicle and entry into theweightless environment. He also noted a false sensation ofaccelerating in a direction opposite to retrorocket firingbefore reentry.

Astronaut Cooper, referring to his Mercury Flight (MA-9,May 15, 1963), has been quoted as having felt

. . . somewhat strange for the first few minutes. . ." afterwhich he "readily adapted" and felt "completely at ease."Later in the mission, following a short nap, he is quoted asawaking "with no idea where I was and it took me severalseconds to orient myself." He also stated that, with respectto sleep, "you have trouble regrouping yourself for a shortwhile when you come out of it." He had an experience,similar to Astronaut Carpenter's on an earlier flight (MA-7,May -24, 1962), when the cockpit seemed to be "somewhatdifferently located in respect to myself," upon onset of theweightless state. He felt, during the early part of the firstorbit, a moving forward in the seat in spite of tightly fastenedrestraint straps, and the equipment storage kit on his rightseemed at a different angle relative to him than when on thelaunch pad. He felt that he was sitting upright although helater described a feeling of hanging upside down because ofpressures against his shoulder straps.

The sensation of hanging upside down from their restraintstraps was also noted by Russian Cosmonauts Yegorov andFeoktistov (VOSKHOD 1, October 12, 1964) but weresimilarly brief and always disappeared, upon the onset ofreentry acceleration. Cosmonauts Gagarin (VOSTOK 1, April12, 1961), Titov (VOSTOK 2, August 6, 1961) and Popovich(VOSTOK 4, August 12, 1962) also briefly experienced thisinversion illusion during their space flights but thephenomenon was reported to have had no effect on theirperformance.

A more significant feature of early Russian spaceflightexperience was the apparent motion sickness syndromereported by Titov during VOSTOK 2. After five or six orbitshe noted symptoms of decreased appetite, giddiness, andnausea which were aggravated by sharp head movements andreduced by keeping his head still. In spite of the fact that the

VestibularAreas of In-vestigation

vehicle was probably rotating slowly and that repeated headmovements combined with anxiety and the initial inversionexperiences could theoretically account for the problem, theresults were interpreted as from a direct"otolithic-vegetative" disorder. Subsequently, a great dealmore investigation and training was devoted to thelabyrinthine and pro prioceptive systems in the Soviet spaceeffort with somewhat ambiguous results. Symptoms asdramatic as Titov's were not again reported until the flightsof Apollo 8 and 9, although temporary motion sickness wasapparently experienced in a milder form by both Feoktistovand Yegorov. The eight and fourteen days of Gemini V(August 21, 1965) and Gemini VII (December 4, 1965),respectively, produced no such symptoms in the four U. S.astronauts involved, although they experienced an increasedgravity sensitivity during retrofire and reentry. This apparentform of increased vestibular sensitivity disappearedpostflight.

Finally, the nausea and vomiting that occurred during theApollo 8 and 9 flights was almost certainly caused byvestibular system malfunctions. The relationship betweenthese symptoms and the greater freedom of intravehicularmovement in the Apollo CM deserves careful attention.

Because of the inability to produce conditions of prolongedweightlessness on Earth, at least two major areas Df vestibularinvestigation are necessary for the qualification of man inlong duration space flight:

1) Ensure that permanent otolithic changes attributable tospaceflight conditions do not occur; .

2) Ensure that temporary vestibular disturbances will notoccur inflight that will interfere with crew safety andmission success.

These areas are also significant to the study of man'saaaptation to rotating space vehicles and artificial gravitytechniques.

The experimental approach in Skylab uses a healthy,well-trained subject adapted somewhat to the unusualvestibular stimuli he has experienced as an aviator in theflight phase. Control groups have been selected so as toinclude some known vestibular defectives. In one test, themeasurement of angular acceleration threshold, a rotatingchair device will be used to provide the subject stimuli ofphysiological character, at best unusual in a subgravityenvironment, and a test goggle to measure oculogyral illusion.Oculogyral illusion, the most sensitive indicator of threshold,is a form of apparent motion that has its genesis in thecupula-endolymph mechanism and may be viewed undermany different circumstances. The test goggle used provides

Labyrintha system ofintercommunicating systems orcanals, such as the inner ear

Propri oceptive receivingstimulations within the tissues ofthe body

55

ReflexVestibularDisturbances

56

the favorable conditions of dimly lighted three-dimensionaltarget viewed in darkness and fixed with respect to thesubject. The expected apparent motion of the target is in thedirection of accleleration.

In another part of the experiment, provocative tests will serveto evaluate the subject's susceptibility to reflex vestibulardisturbances and to motion sickness and may, in addition,measure his ability to cope with such disturbances. Thesetests will measure the coriolis sickness susceptibility indexusing, again, the rotating chair but with the subject makingstandardized head motions out of the plane of rotation.

A unique feature of this test is the method of scoring theinvestigators have developed. This highly effective methodyields a single value, the index, enabling the investigator tomake comparisons within and among test subjects. The levelof severity of acute motion sickness will, therefore, beabsolutely controlled to a definite end point defined asMalaise IIA. (See Table 6-1). The test end point will be fixed,while the level of stresses required to reach that end pointwill be measured (number of sequence of head motions androtational velocity).

Table 6-1 Diagnostic Categorization of Different Levels of Severityof Acute Motion Sickness

Category

Pathog-nomic16 Points

Major8 Points

Minor4 Points

Minimal2 Points

AOS*1 Point

Nauseasyndrome

Skin

ColdsweatingIncreasedsalivation

Drowsi-ness

Pain

Centralnervoussystem

Vomitingor retch-ing

Nausea+II, III

PallorIII

III

III

III

Nausea 1

Pallor II

II .

II

II

Epigas-tric dis-comfortPalor I

I

I

I

Epigastricawareness

Flushing/sub-jective warmth...- II

Headache >IIDizziness, eyesclosed ..>- II,eyes open III

FrankSickness

Levels of Severity Identified by Total Points ScoredSevereMalaise

ModerateMalaise A

ModerateMalaise B

SightMalaise

(S)?.-- 16points

(M III)

8-15points

(NI IIA)

5-7points

(M IIB)

3-4points

(M I)

1-2points

*AOS = Additional qualifying symptoms I = moderate,+III = severe or marked, II = slight.

Epigastricpertaining to theupper middle region of theabdomen, located within thesternal angle

SpatialLocalization

The spatial localization experiment uses the rotating litterchair in the static tilt and litter modes, together with theotolith test goggles, blindfold, and a reference sphere andmagnetic pointer. The astronaut's capability to determine hisorientation is tested with the chair upright as well as tilted.

The subject is first tilted to various positions relative to thespacecraft with his eyes closed and is asked to indicate bothhis perceived direction of gravity and body orientation. Heindicates this both by setting the direction of an illuminatedline in the test goggles and by lining up a magnetic indicatorrod on a handheld sphere.

In the litter mode, the reference sphere and magnetic pointerand blindfold are used. Tests will investigate orientationbased upon a sensed g-vector provided by external referencecues and internal reference cues. The chair is converted to alitter and the subject attempts to align the magnetic pointerto an external reference while blindfolded. Settings are madewith the litter in a horizontal position and with the litter in atilted position.


The objectives of this experiment are to acquire data tovalidate measurements of specific behavioral andphysiological responses as influenced by vestibular activityunder Earth gravity and in free-fall conditions. These studiesare also expected to determine the crewman's adaptability tounusual vestibular conditions, and to predict habitabilityrequirements for future spacecraft which may be rotated togenerate an artificial gravity force leading to unusual coriolisforces. Another objective is to measure the accuracy andvariability in the crewman's judgment of spatial coordinatesbased upon abnormal receptor cues and inadequate visualcues.

EQUIPMENT

The equipment for this experiment consists of --

1) rotating litter chair,2) proximity device,3) blindfold,4) otolith test goggle (with a biteboard mounting),5) reference sphere with a magnetic pointer.

The rotating litter chair is a precision motor-drivenservocontrolled, rotating chair which can be tilted in twodirections or converted to a litter; however, it is only rotatedin the upright position.

Coriolis Forcesa fictitiousforce used to describe motion(aircraft or cloud formation)relative to a noninertial,uniformly rotating frame ofreference

57

MotionSensitivity

58

The right eyepiece of the otolith test goggles contains aself-illuminated target with pitch and roll adjustment. Thetarget is a slit of light that can be rotated to indicate a rollposition; a break in the middle indicates the pitch position;and a second small break at one end indicates polarity. Thetarget, the only thing visible to the subject, is illuminated bya radioactive source in the goggles.

The otolith test goggle is fastened to the chair so that whenthe subject bites on a biteboard and the goggles are properlyadjusted, the head will be held in the correct position for thetest.

The reference sphere with magnetic pointer is a device formeasuring spatial localization using nonvisual clues. Amagnetic pointer is held against the reference sphere andmoved by the subject. The subject's judgments are measuredby a three dimensional readout device that allows freetranslational movement.

PERFORMANCE

Each of two crewmen will participate in dynamic vestibulartests consisting of motion sensitivity tests using the rotationlitter chair in the rotating mode to determine susceptibilityto coriolis forces as a function of time in weightlessness, andto measure semicircular canal response thresholds byconducting oculogyral illusion (OGI) threshold tests. Thissequence will be done on 5 equally spaced occasions duringthe first 28-day mission. Each test sequence requiresapproximately 30 minutes per crewman.

Each of three crewmen Will participate in spatial localizationtests using the otolith test goggles and the rod and spheredevice. These tests will be performed once early in themission, once in the middle of the mission, and once late inthe mission.

DATA

During motion sensitivity tests:

1) test subject symptomatology in accordance with Figure6-1 for determination of time course of symptomdevelopment to the test end point;

2) experiment data pertaining to chair velocity, direction,head motion characteristics, and experimental equipmentperformance.

During oculogyral illusion tests:

1) oculogyral illusion response;

0 c ul ogyralpertaining tomovement of the eye about theanteroposterior axis

SleepMonitoring

2) stimulus characteristicschair acceleration, velocity anddeceleration;

3) experimental equipment performance data.

During spatial localization tests:

1) accuracy of subjects spatial localization and testconditions (subjects actual orientation relative to thespace labs spatial coordinates).

Motion picture films of the experiment performance in allmodes will be obtained.



One of the most dramatic and fundamental ways in whichthe behavior of a mammal can change is in its state ofconsciousness. There is a spectrum of changes from alertnessthrough quiet resting to drowsiness to various kinds of sleep.These changes in states of consciousness affect the activity ofthe nervous system in many ways. In the last few years ourunderstanding of states of consciousness has increased (andchanged) markedly.

One way of examining what is going on in the brain is bymeans of an EEG (electroencephalograph). The EEG is arecording of the electrical activity from the cerebral cortex.It is clear, that the EEG is not just due to action potentials incells, although these probably contribute. Synaptic currentsplay the largest part. There are good correlations between theEEG and states of consciousness.

During alert, attentive activity the EEG is "low voltage, fast"i.e., waves of less than 100 microwatts recorded from thesurface of cortex with frequencies mostly greater than 15 Hz.When the subject is in a relaxed state, conscious but notattentive to anything, the EEG has an alpha rhythm. Thealpha rhythm has a frequency of 8 Hz and amplitude of up to400 microwatts. As the subject becomes more drowsy, thefrequency becomes slower. This is mixed with bursts ofelectrical activity at 10 to 14 Hz lasting a few seconds. Insome kinds of sleep, the EEG waves are 2 to 3 Hz and about400 microwatts. However, in another kind of sleep, the EEGis low voltage, fast, almost exactly as in the alert state. Allmammals demonstrate this same pattern although the exactfrequency is somewhat different from animal to animal.

The arousal mechanism subserves attentive alert behavior andthe associated EEG pattern. There is an anatomical site that

Synapticpertaining to theanatomical relation of one nervecell to another

59

ParadoxicalSleep

60

governs this activity, the reticular formation of themesencephalon (in the upper brain stem) and the adjacentposterior parts of the hypothalamus. This is sometimes calledthe reticular activating center. Electrical stimulation of thissite causes the animal to become alert, a response that willlast longer than the duration of the stimulus. Destruction ofthis center produces coma. Among the normal inputs to thiscenter are those from all the sensory systems. A single cell inthis center may be fired by a flash of light, a ringing bell, atouch on the skin, or a smell. It is polysensory. This makessense in terms of the function of the center, for any kind ofsensory stimulation, if sufficiently intense, will cause ananimal to become alert.

There are many aspects of arousal. The musculature increase5its tone, and the cardiovascular system is affected. Witharousal the sensitivity of most aspects of sensory systems isdecreased. This may seem paradoxical, but this eliminatesirrelevant inputs and presumably allows greater attentiontoward specific sensory inputs. There is no simpleinterpretation of the effects of arousal on the activity of thecerebral cortex. Some cells fire more, some less; some aremore excitable, some are less excitable.

There are two kinds of sleep. One kind, which we will callslow wave sleep, comprises about 75% of sleeping time. Theother kind of sleep, which we will call paradoxical sleep, inhumans comes in episodes of 10 to 20 minutes durationevery 30 to 90 minutes and comprises about 25% of sleep.(Paradoxical sleep is also called rapid eye movement sleep orREM sleep.) Paradoxical sleep can only be reached by goingthrough slow wave sleep.

Paradoxical sleep is very stereotyped. It starts abruptly, itinvolves a loss of tone in muscles, except those of the faceand eyes. The facial muscles twitch and there are rapidmnvements of the eyes. The blood pressure decreases. TheEEG is low voltage fast frequency. If a human is awakened,especially during the early stages of deep sleep, he willusually report a dream. If an animal is awakened as soon asan episode of paradoxical sleep begins, another episode ofdeep sleep cannot start again for at least 30 minutes. Ifanimals are awakened whenever they begin to have anepisode of deep sleep, because of this "refractory period,"they can be prevented from spending much time inparadoxical sleep, although their total sleeping time remainsnormal. All animals, after being deprived of paradoxicalsleep, show an increased proportion of paradoxical sleep thenext time they sleep.

A center for paradoxical sleep has been found in the pons(part of the brain stem). Stimulation of this site initiatesparadoxical sleep; destruction of the site prevents paradoxical

Muscul turethe muscularapparatus of the body, or anypart of it

Slow WaveSleep

sleep. Norepinephrine, which is presumed to be a synaptictransmitter substance, is concentrated in these areas andappears to be very much involved in paradoxical sleep.

Slow wave sleep (non REM or NREM) can be initiated bystimulation of many parts of the brain. Many other sitesinitiate slow wave sleep if stimulated at about 8 Hz, even aperipheral nerve. Slow wave sleep, unlike arousal andparadoxical sleep, seems to require that the cerebral cortexbe present. However, a major contribution to slow wave sleepcomes from the medulla (part of the brain stem). Serotonin, Serotonina constituent ofwhich is presumed to be a synaptic transmitter substance, is blood plateletsconcentrated in these areas and appears to be very muchinvolved with slow wave sleep. If a barbiturate generalanesthetic is injected into the blood going to the posteriorbrain's stem (i.e., to the pons and medulla) the animal willwake up. If the anesthetic is allowed to go to the moreanterior part of the brain the animal will be anesthetized,which is similar to slow wave sleep in many ways. Slow wavesleep and alert behavior are part of a continuum. There aremany intermediate stages. Paradoxical sleep, however, is allor none, with no intermediate stages.

It would appear that there are three active mechanismsinvolved in states of consciousness. Sleep is not just theabsence of consciousness.

The blood flow and oxygen consumption of the brain is thesame in arousal and slow wave sleep, but is increased inparadoxical sleep. There have been a few experiments inwhich recordings have been made from the same nerve cellwhile the animal was spontaneously awake, and then asleep.Some neurons fire more during sleep, some less. Mostneurons fire more during paradoxical sleep. Therefore, thebrain does not rest in sleep. In fact, it is most active inparadoxical sleep, and it is resting when it is awake. Thecommon sense view and the scientific view until about 1963was that the function of sleep was to rest the brain. Todaywe are not certain as to what the function of sleep is, but wedo know that it is essential to physical and psychological wellbeing.

From the beginning of the space program there has beenconsiderable interest in whether the astronaut was gettingadequate sleep. In order to answer this question NASA hasbeen investigating the use of the EEG for inflight monitoringof sleep. These investigations include determination of theminimum number of brain areas from which signals that yielduseful information are picked up, and whether an onboardcomputer can be used for performing the analysis, thusreducing the complexity of the telemetry system that isneeded to return data to Earth. A preliminary approach to

61

MonitoringSleep ofAstronauts

62

the problem was undertaken to 1965 and continues to thepresent time under the direction of the Principal Investigatorfor this experiment.


The primary goal will be to monitor the sleep status of aspacecraft crewmember during selected periods throughoutan extended space mission, utilizing automatic onboardanalysis of electroencephalogram (EEG) andelectro-oculograph (EOG) with telemetry of results. Sleepprofiles will thus be available in the Mission Control Centerand may be readily evaluated in order to detect anyalterations in sl,:p quantity or quality.

EQUIPMENT

The major components of the experiment equipment are:

1) cap assembly,2) preamplifier and accelerometer assembly, and3) control panel assembly.

The cap assembly precisely fits the astronaut's head andcorrectly positions seven electrodes, which are part of it. Theelectrodes are funnel-shaped, soft "rubbery" devicescontaining electrolyte soaked sponges. The lower portion ofeach electrode is cut off by scissors just before the crewmanputs on the cap, thus exposing the electrolyte for propercontact with the scalp. A new cap will be supplied for eachexperiment sleep period.

PERFORMANCE

Three consecutive nights of sleep recording will be requiredof the prime and backup crewmember within 60 days beforelaunch, plus a one-hour session in the Principal Investigator'slaboratory.

A total of 15 selected sleep periods shall be recorded duringthe first (28-day) mission, and 21 sleep periods during thesecond mission (56 days).

Three nights of sleep recording are desired on postflight days1, 3, and 5, using the same procedures and basic equipment.

The procedure for this experiment is quite simple. On thespecified nights, before retiring, the astronaut takes a capassembly, cuts the tips off of the electrodes, connects the capassembly to the rest of the equipment, dons the cap, andsecures himself in the sleep restraint. After checking theoperation of the equipment he goes to sleep in a normal

manner. As he sleeps the equipment automatically performsthe analyses and provides data to the telemetry system. Uponawaking he disconnects the equipment, throws away the cap,and makes any comments needed in the log and to the voicedata system.

DATA

1) Copies of all data tapes obtained during preflight andpostflight baseline testing will be provided to thePrincipal Investigator for analysis.

2) A log entry is required after each monitored sleep period,preflight, inflight, and postflight. This entry, made byvoice recording, will contain information on the durationof sleep, quality of the sleep, and any medication used bythe subject with 24 hours before the beginning of themonitored sleep period.

63

i

Section 7

Biology

Effects of Zero Gravity on Single Human Cells,Skylab Experiment S015

Circadian Rhythm-Pocket Mice,Skylab Experiment S071

Circadian Rhythm-Vinegar Gnats,Skylab Experiment S072

BiologicalProcesses

GENERAL BACKGROUND

Many biological processes of living organisms are periodic innature. The most well known of these are the circadian(daily) rhythms of sleep, body temperature, flower petalmovements, etc, but there are many other biologicaloscillatory frequencies as well. These rhythms are believed tobe evolutionary in origin and responsive to terrestrial cuessuch as light/dark cycles. During manned space travel beyondhis evolutionary environment, temporal factors will have tobe included as part of the provided environment to avoidbreakdown of periodicities. Although work/rest schedulesmay be the governing input, there are basic biologicalquestions related to circadian rhythms that may be ofimportance.

It is possible to divide circadian rhythm research into four Circadianpertaining to a periodcategories: (1) occurrence and general characteristics, (2) of about 24 hoursenvironmental phase-setting, (3) the basic mechanism, and(4) the importance of these rhythms to the organism.

To summarize briefly: (1) from unicellular organisms to man,there are few biological processes that do not demonstrateperiodicity; and (2) circadian processes have a labile temporalrelationship with the environment and can be phase-set(entrained) with appropriate light or temperature stimuli.

The latter two aspects, however, have evaded scientificinquiry, although many investigators are attempting todetermine the mechanism of circadian rhythms and thephysiological consequence of their disruption.

Circadian rhythms, rather than being direct biologicalresponses to the immediate temporal environment, may servea more basic role in the functioning of living systems.Processes must occur not only at the proper place and insufficient quantity, but must also occur at the propertime. Thus, the circadian mechanism permits intervalsynchronization of an organism's processes and at the sametime synchronizes the total organism to its environment.

A review of the literature on human circadian rhythms showsover 50 physiological variables with significant circadianvariations. Although this variation is only a small percentageof the total change for a function such as vital capacity, itaccounts for the majority of the variation in others such aselectrolyte excretion. The circadian changes observed areoften caused by variations in other processes. Thus, it is notalways possible to determine the direct relationship betweenthe observed rhythm and the basic causal mechanism.

Changes in circadian rhythms may occur during certainpathological conditions. In addition, there may be decreased

65

Circad ianRhythmTheories

Effects ofZero Gravityon Single Hu-man Cells

psychological performance associated with changes Dysrhythmiadisturbance of(dysrhythmia or desynchronization) in circadian rhythms. rhythm

This phenomena is becoming a well-known effect of rapidtravel across several time zones.

There are two theories regarding the basic mechanism ofcircadian rhythms: (1) they are the result of a completelyendogenous mechanism (biochemical or biophysicaloscillator) with other environmental factors, such as light ortemperature, acting as phase - setter;,, and (2) they representan organism's response to subtle geophysical fluctuations(such as magnetic fields) with light and temperature againacting as phase-setters between the basic mechanism and theenvironment.

The majority of investigators subscribe to the first theory;however, the required critical evidence has not beencollected. The Skylab Program offers an environment inwhich the two hypotheses can be tested, since during orbitalflight the geophysical variations are no longer available to theorganism on a 24-hour basis, and during interplanetary flightthey should be completely absent or greatly distorted.

At present, there is only suggestive evidence of detrimentaleffects due to disruption of normal rhythms in abnormaltemporal environments. To date, it has not been possible tocompletely abolish circadian rhythms, only to perturb them.If the second hypothesis proves to be true, a completely newfactor will have to be considered during long duration flights,i.e., the lack of a basic temporal coordination mechanism.

SKYLAB EXPERIMENT 5015


A great deal is known about the behavior of human cells in a1-g environment; both normal and abnormal behavior is wellunderstood. However, virtually no information exists on thebehavior of cells in a near weightless environment.

This experiment to study the influence of weightlessness onliving human cells is designed to obtain information on celldevelopment and propagation by using two different andseparate methods.

Observation of The first method will be through observation of two livingCells cells using microscopes and cameras. These cells will be kept

alive for the ,entire mission and returned alive. The cellbehavior will be periodically recorded during the mission.This technique, using phase contrast microscopes (specimendoes not have to be stained) and time-lapse photographs willallow, postflight study of the visible portions of a cell.

66

ChemicalPropertiesof Cells

Examples of this will be changes in cell size, cell division(mitosis), and changes in size and movement of specificparticles in the cell (organelle). Kidney cells will be usedbecause they flatten out in the specimen chamber so that thenucleus and other particles are easily seen under a low powermicroscope.

The second method will deal with the chemical properties ofcells and will be conducted during periods of rapid growthrate. This portion of the experiment will be completed on thefourth and tenth days of the mission, at which time the cellsare fixed and will be held in that condition until they arereturned for ground analysis.

The cell chemical study will use 24 separate em ryonic lungcell cultures. The cells will be allowed to grow :or four daysand ten days, respectively, after the start of the mission. Onthese days various radioactive materials that can beassimilated by the cell will be introduced into some of thespecimen chambers at specific times and for specificdurations. This will mark or label various functions of thecells at that particular time. After the cells have been labeled,excess radioactive material will be rinsed from the chamberand all of the cells preserved with a fixative that will holdthem in that state until they are returned for ground analysis.

The information that will be gained from this portion of theexperiment will be (1) the distribution of chemicalcomponents in the cell (histochemical), (2) the amounts and Histochemicalchemistry ofrates of synthesis of various chemical constituents, and (3) tissues

the ultra structure, or arrangement of the very smallestelements of a cell.


This experiment will determine effects of liear weightlessnesson living human cells in a tissue culture.

EQUIPMENT

The experiment package consists of two major subsystemsboth enclosed in a single hermetically sealed packagethemicroscope-camera subsystem and the biopack subsystem.See Figure 7-1 for a simplified block diagram showing thefunctions of the experiment.

The biopacks and the microscope-camera subsystems arecontained in a hermetically sealed experiment package whichalso includes two clocks that control the automaticoperations. The two biopacks are enclosed in anotherhermetically sealed container within the experiment package.

67

Human CellExperiment

68

Camera microscope- Speciman MediaControl

H4OXcamera chamber pump

20X microscope- Specimencamera chamber

j. Motoranddrivetrain

Biopack 1

Media Label Rinse Label Rinse Fix

II .1141.LSelector valve

11112111111111111Specimen e eE a arm FA. e ,E,chambers

4 Biopack 2

Pressure Shell

Figure 71 Simplified Schematic of Human Cell Experiment

The 20X and 40X power microscopes shown in Figure 7-1have exposure lamp assemblies that project an image down alight-tight tube to a mirror and film gate assembly. The imageis reflected on the film or in a viewport for ground checkoutwhen the mirror is manually rotated 90 degrees. The viewingmechanism is used only for ground checkout and isspring-loaded to prevent the mirror from being inadvertentlyleft in the viewport position after ground checkout. Nofurther adjustments are made in flight.

Each camera assembly operates independently and iscontrolled by separate internal clocks. Both cameras use16mm film for recording microscope images of the cells.

The two camera assemblies operate identically. One cameraphotographs time-lapse motion pictures of images from the40X magnification microscope. The second cameraphotographs images from the 20X microscope. A removablefilm pack is used to contain the 16mm film that is requiredfor each camera. The two films travel in opposite directions,thereby maintaining a constant center of gravity.

Two separate specimen chambers, one for each microscope,will provide a temperature-controlled environment in whichthe cells can live.

Each specimen chamber has an independent media exchangeassembly to provide fresh nutrients to the living cells. Twice

Time-Lapse Photographya slowand continuous process atregular intervals

CircadianRhythmPocket Mice

each day (not concurrently with the photographing period)fresh nutrients are forced into the specimen chamber andwaste removed.

Each of the two specimen chambers contain two thin glasscoverslips with a molded rubber gasket held between them.The specimens are attached to the inside of the coverslipnearest the objective lens. Two tubes are attached to thecoverslip for supplying nutrients, and removing wastematerial from the space between the coverslips. Eachspecimen chamber has a media pump assembly. This pumpconsists of a cylinder, a piston and a lead screw. Twice dailythe lead screw is turned one revolution to advance the pistona short distance into the cylinder. This action forces freshmedia into the specimen chamber and waste media to thereturn side of the pump.

The biopack subsystem consists of two independentassemblies enclosed in a sealed container inside theexperiment package. Each assembly controls 12-cellchemistry experiments and each assembly contains 12 cellchambers, a motor drive assembly, a pump for providingfresh nutrients and removing waste from the cells, andprovisions for labeling, rinsing, and fixing the cell cultureswhenever commanded by a crewmember.

PERFORMANCE

The filming of the living cells is performed automatically.The ,:rewman has to check only for the proper operation ofthe camera indicator lamp for the first 10 days. In the otherpart (chemical properties) of the experiment, the crewmanwill operate the experiment manually at the 4- and 10-daypoints to perform the radioactive fluid labeling activities.

During the mission, the experiment package must not besubjected to temperatures above 95°F or below 50°F, or toradiation x-ray sources strong enough to damage the film.

DATA

Time-lapse motion pictures of the cell activity, as seenthrough the 20X and 40X magnification microscope, will bemade and the results of the cell chemical analyses will bepublished in study reports, possibly early in 1975.



If the stability (precision) or the period of physiologicalrhythms of small mammals change significantly during flight,then there is a strong indication that biorhythms of animals

69

Body Tem-perature andActivity

70

on Earth are timed by some factor (or environmental force)which is absent or significantly altered in space. This changein the behavior of pocket mice, along with similar evidencefrom the vinegar gnat circadian rhythm experiment, wouldimply that weightless spaceflight alters the functioning ofbasic control mechanisms for metabolic activity. Themaintenance of normal biological rhythms in man duringspaceflignt is important to his well-being and effectiveness inspace. Insofar as we can assume a common response inmammalian function, if the pocket mice in space continuetheir terrestrial biorhythms, then we can conclude that spaceconditions impose no stress on the basic biological clockmechanism and that man's performance will not be degradedbecause of rhythm disturbances.


The objective of this experiment is to study the circadianrhythms of body temperature and activity in pocket mice( genus, perognathus) in an environment of constanttemperature, pressure, and total darkness, and in the absenceof geophysical variables within a 24-hour period.

EQUIPMENT

The experiment consists of a sealed animal enclosure coupledto an environmental control system. The animal enclosurecontains six isolated mouse cages, the biotelemetry receivers,and monitoring detector circuitry. The environmental controlsystem maintains the mice in a controlled environment ofconstant temperature, pressure, and darkness.

The cages are cylindrical tanks with porous sealed covers andhave an internal height of 1.5 inches and a floor area of 18square inches. Each cage contains food for the mice. All thecages receive equal ventilation from a common air supply.Figure 7-2 shows a cross section of the cages. The inside ofthe cage except for the fiberglass tubing is covered with alayer of porous material.

The environmental control system is illustrated in Figure 7-3.The environmental control system tank houses a fan tocirculate the air, a charcoal-lithium hydroxide absorptioncanister to remove carbon dioxide and ammonia, a dew-pointcontrol heat exchanger, a moisture separator, and

ttemperature contro heater. The accessories mounted on thetank include an ox gen supply tank and regulators, the fluidpump control elements and plumbing for connecting the heatexchanger to an external cold plate. The entire experimentpackage is surrounded by a multilayered blanket ofaluminized Mylar.

A biotelemeter with a self-contained power source isimplanted in each mouse to monitor and transmit body

Biotelemetrythe recording andmeasuring of certain vitalphenomena of living organisms,occurring at a distance from themeasuring device

Heaterelement

Shield screen

Foam

Orifice

Foam

Air inlet

Subplenum

Housing

4111111°a

/=.-

Mouse cagecavity

Figure 7.2 Cross Section of Cage

Plenum

Air outlet

Antennas

Inlet

To mousehouse, airtemperaturecontrolled

Mouse fCage

Mouse \Cage

Mouse Cage Mouse 7.."FCa e

Plenum Heater controlsensor

Mouse.- Cage Mouse '

Lo pressureregulator

Hi pressureregulator

02 02 chargingtank valve

-02Outlet

.411-

Moistureseparator

Figure 7-3 Environmental Control System

Outlet

Heatexchanger Li-OH

.C-N. /is N. 12AN b-Gas absorption Fancanister froActivatedcharcoal 2 typs

--/

t 1To supplementalcooling system

71

CircadianRhythmVinegarGnats

72

temperature information. The animal activity is determinedfrom variations in the signal strength as the mouse changesposition with respect to a receiving antenna in the center ofeach cage. The data acquisition and processing is done by theelectronics data system.

PERFORMANCE

The experiment is performed during 28 days of the mission.Before launch, 24 pocket mice are placed in a simulatedflight hardware environment and maintained in a constantenvironment for at least three weeks to document the "freerunning" circadian periods of body temperatures and activityof each mouse. Two groups cf six mice are selected from the24 and placed into the two flight units following the baselinerun. One of the flight units is placed into the launch vehiclewhile the remaining flight unit is used as a ground control.The circadian rhythms of each mouse are continuouslymonitored from launch for 28 days minimum whilecontinuously maintaining the constant internal environmentof the flight hardware.

Additional groups (those remaining from the 24) of pocketmice are maintained in a constant environment at the launchsite, and monitored simultaneously with the flight group forbody temperature and activity.

DATA

The measurements from this experiment are taken for theduration of the mission. Data will be transmitted to theground data stations every 8 hours. Analyses of the data fromthe flight and ground control mice will be reported in ascience report published about a year after the mission.



The first formal record of biological rhythms was probablymade by the astronomer, De Mairan, who in 1729 describeddiurnally periodic leaf movements in plants held in the dark.The literature is now replete with evidence of diurnalperiodicity in many forms of life at many levels of Periodicitytendency to recur atorganization. Present day research emphasizes on one hand, regular intervals

the mechanisms of the so-called "biological clock," and onthe other, the coupling of the clock to environmental stimuli.Whether control of the period of the clock is predominantly aphysiological or environmental phenomenon is debatable onthe basis of existing data. Theoretically, the study ofbiorhythms in space would permit resolution of the questionas to whether terrestrial stimuli indeed set the period orsimply determine its phase.

Effect of ZeroGravity onCircadianRhythm

The principal evidence for external stimuli setting the periodof the circadian rhythm is the (1) remarkable persistence andprecision of the rhythm in organisms kept in constant.vironments apparently free of stimuli that are capable ofentraining a self-sustaining oscillation, and (2) the remarkableinsensitivity of the free running period to the level oftemperature (temperature compensation). If the control ofthe period were truly endogenous (metabolic), changes intemperature should have produced pronounced changes inperiod in poikilothermic organisms. On the basis of theseobservations, Dr. Frank Brown of Northwestern Universityhas postulated a precise 24-hour component in the circadianrhythm resulting from entrainment to some "pervasivegeophysical force."

Most workers, however, believe circadian periodicity to beprimarily inherent in the organism. Recent evidenceimplicates a role of genetic structure in setting the period,loci having been found which can effect an aperiodicity. TheSkylab experiment is an effort not so much to provide adefinitive answer to the opposing contentions as to shed lighton the effect of zero gravity and removal from the Earth'simmediate geophysical parameter on a precisely measuredbiological phenomenon.

Drosophila eclosion rhythm is the best studied circadiansystem of any organism. Techniques fully developed atPrinceton University permit the selection of a pupalpopulation that would emerge in, e.g., three or foursuccessive peaks of activity separated by a precise circadianperiod. Methods of assaying the emergence of the adults inthe weightless state and providing the environmental controlwhich the experiment requires have been under study atNorthrop Corporate Laboratories.


The objective of this experiment is to study the circadianrhythms of vinegar gnat pupae (genus, drosophila) underconditions of weightlessness and in the absence ofgeophysical variables within a 24-hour period associated withthe Earth's rotation.

EQUIPMENT

The equipment consists of four pupae compartments, thecircadian data system and power supply, and the controlelectronics.

Each cylindrical scaled pupae compartment consistsessentially of a control post that supports 180 pupae on aplate, scanning lamps, photosensors, a stimulus light, a to light

Drosophila the fruit fly, usedextensively in experimentgenetics

Photosensora device sensitive

73

EiperimentInvestigation

74

solution of potassium hydroxide, and environmentalControls. The pupae plate contains 12 beds equally spacedaround its periphery. Each bed contains 16 holes. Pupae areglued over 15 of the holes so that they completely obscurethe holes; the 16th is uncovered to monitor the scanninglight. One hundred ninety-two photosensors are mountedunder the pupae plate, one for each hole. Twelve of thesephotosensors monitor the scanning lights.

Each of the 12 beds has an overhead red scanning light thattransmits through the pupae onto their individualphotosensors. The detection system can discriminate betweenan empty pupae case and developing pupae which exhibitvarious degrees of transparency during development, but onlyafter eclosion is sufficient light transmitted through theempty pupae case to activate the detection system. The bedsare scanned sequentially for 0.5 seconds every 10 minutes.

The sealed compartments are charged with air of specifiedcomposition to maintain the pupae throughout theexperiment. The initial charging pressure is adjusted so thatwhen the compartments are heated to their experiment level,the pressure will rise to sea level pressure. The carbon dioxideconcentration and humidity in the compartments arecontrolled by a potassium hydroxide solution. The internaltemperature is regulated to automatically supply or removeheat.

A white light provides a stimulus with a wavelength between400 to 700 nanometers to the pupae to determine the phaseof the biological clock controlling eclosion.

PERFORMANCE

The experiment is performed during the first 20 days of themission. All crew operations are to be performed only upondirection from the ground flight controllers.

In the circadian rhythmvinegar gnat experiment, apopulation of 720 vinegar gnat pupae divided into 4 groups isplaced in Earth orbit in a dormant state. When in orbit, thepupae are warmed to approximately 20°C to allowdevelopment. Sometime after the temperatures are stabilized,the pupae are exposed to a single 2-minute pulse of whitelight. The light pulse sets the phase of the circadian rhythmsand establishes the baseline from which the circadianrhythms are determined.

The eclosion of pupae (emerging from puparia) in each of Pupariapupal cases formed ofthe compartments is continuously monitored until the cuticula of preceding larvalexperiment termination. instars

In addition to the inflight pupae, an equal population ofpupae will be housed at the launch site. These pupae will be

stimulated simultaneously with the flight group, exposed tothe same controlled environment, and monitored for thesame data.

The data acquired from the experiments will be analyzed todetermine any variance among the circadian rhythmsobserved in the flight group while in orbit and the circadianrhythms observed during the baseline run and those observedin the ground control groups. A significant digression of'either the precision or length of the free-running circadianperiods measured in space from those periods measured onEarth would constitute evidence of dependency of circadianorganization upon conditions of spaceflight. Conversely,continuance in space of the precise free-running circadianperiods measured on the ground would constitute evidence infavor of the independence of circadian organization ofspaceflight conditions, including weightlessness.

DATA

The data from this experiment are recorded from prelaunchuntil deactivation of the experiment. Data will be transmittedto the ground data stations every 8 hours. Results of theanalysis of this data will be published in a science reportpossibly early in 1975.

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Section 8Classroom Activities

i

In preceding sections a fundamental tie has been establishedbetween the Skylab experimental program and the sciencethat guides and disciplines man's quest for knowledge abouthimself. In this section, a few classroom activities, discussion

...topics and student experiments which the teacher mayemploy to demonstrate the principles and techniques ofscientific inquiry are suggested. These simple qualitativeclassroom activities can be coupled with more precise andquantitative data and aids from Skylab to enhance theeducational process. The suggested demonstrations thatrepresent only a very small sample of those which might beperformed, have been included only to show the scope ofrelevant experimental techniques.

CLASSROOM ACTIVITIES

Body Mass The evolution of the mammalian skeleton has beensignificantly influenced by factors such as gravity, which isthe primary discussion topic of this section.

The weight of an animal is proportional to the gravitationalattraction and the mass of the body. The relative mass of abody is directly proportional to its volume. A very largesphere has much less surface in proportion to its volumethan a very small sphere: If the radius of a sphere is increased,its surface increases by the square (A = 47rr2 ), but its volumeincreases by the cube (V = 4/3/r? ).

The following discussion is adapted from DuBrul*

The operation of the principle that volume of a sphereincreases by the cube may be applied in different ways in theanimal world. Suppose we pretend that the body of ananimal is like a sphere and that four legs support this sphere.The four legs are subjected to pressure from the weight of thebody. What is the relationship of the thickness of an animal'slegs to volume of its body? A horse has long thin legs. Amoose is larger than a horse, but its legs seem to be only alittle thicker. An elephant is still larger, but its legs areenormously thicker. Now we are near the upper limit of sizefor living land animals. Not only is the proportion of legs tobody of these three animals different, but the locomotion isalso different. The horse has a rapid gallop; the moose aslower pace; the elephant a steady plod.

The African elephant can weigh as much as 61/2 tons. With itsbone structure this elephant is near the limit of its size, whilestill retaining its mobility. However, a whale, the largest of allmammals may weigh forty times as much as an elephant. The

*DuBrul, E. Lloyd: Biomechanics of the Body, BSCSPamphlet #5, January, 1963, American Institute ofBiological Sciences, pp 2, 3

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The Conceptof Cause

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whale's bones are not proportionately thickened, but theyare strong enough because the whale is supported by water.Many of the dinosaurs approached whale-like size. How werethey adapted for survival?

A discussion of one of the problems that would arise with theincrease in size of an organism follows.

If an organism were to double in size, could it retain the samestructural makeup under Earth gravity conditions? Bydoubling its size, an organism would increase eight times involume (weight) with only an increase of four times instrength of the skeleton. (The strength of a bone isproportional to the diameter squared. This scaling restraint isdiscussed more fully in Physics, D.C. Heath & Company,1960 edition, p 45 ff, or in the 1965 edition on page 48 ff.Reading these pages could lead to an interesting discussion onevaluation of organisms under varying gravitationalconditions.

Other interesting problems that could be posed for discussionpurposes are:

Are animals possessing no skeletons (invertebrates)subjected to the same limitations as vertebrates?

How much increase in size or mass would be possiblebefore additional skeletal strength would be necessary ifan organism were transferred to gravity conditions on theMoon (gravity 1/6 Earth's)?

The leg bones of one animal are twice as strong as thoseof another closely related animal of similar shape. Whatwould you expect to be the ratios of the animals' heightsand weights?

Because of the problems associated with conductingexperiments involving the use of hormones, the followingInvitations to Inquiry are submitted for discussion topics.The source of these Invitations is the Biology Teacher'sHandbook (John Wiley, 1970), prepared in cooperation withthe Biological Sciences Curriculum Study. "The teacherpresents the background of the invitations orally and whennecessary at the blackboard.* He then poses the problem ofthe invitation and invites student reaction. Thereafter hedeals with student responses as they arise, asking diagnosticquestions which help students see what is wrong with pooranswers, reviewing the logic that justifies good responses.

Physics, D.C. Heath & Company, 1965 p 48

*BSCS has developed inquiry slides which relate to theretopics and are related to these invitations.

Sound responses to early problems of an invitation then leadnaturally to the next problem in the invitation and theprocedure of diagnostic and analytical questioningcontinues."

INVITATION #17

Subject: Thyroid Action (p 108)Topic: Unit Causes

`One of the easiest of complications to understand is the factthat we can always try to break big causes into little ones. Awhole gland, such as the adrenal, for example, can be treatedas a cause. We remove it and note the consequences ofremoval. Then we interpret these consequences as indicationsof the normal effects of the adrenal gland. There is nothingerroneous about such an experiment and interpretation, butit is certainly incomplete, for we can go on to divide thegland into parts, such as medulla and cortex, and redo ourexperiment using one such part at a time. Later, we mayanalyze still further. Perhaps we woulu proceed bydistinguishing different kinds of cells in the cortex and tryingto ---,move one kind of cell at a timeand so on down tomc .cules.

`Back of this progressive analysis of causes into finer andfiner parts is the idea that somewhere we will arrive atirreducible unit causes, or causal elements. However, thepossibility of such elemental causes does not mean thatgrosser, composite causes, such as whole organs or tissuecomponents of organs, are "wrong" or useless. On thecontrary, they may be very useful, not only in appliedbiology, as in medicine, but also in research.

`Hence, what we want to convey to students is not that onelevel of analysis is better or more "scientific" than another,but only that there are many levels.' (p 108)

Invitation #17 uses this procedure to investigate the action ofthe thyroid gland.

INVITATION #21

Subject: Parathyroid Action (p 116)Topic: Multiple Causation

This Invitation illustrates that a given effect may be evokedby what appears to be several different causes.

`This is sometimes referred to as multiple causation, but theterm is something of a misnomer. It suggests that several

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genuinely different things or events can cause precisely thesame effect. This could be the case in nature. However, theconcept of causation underlying much biological researchinto causes includes the idea that, ultimately, if severalinstances of an effect are identical, the cause will also befound to be the same.

`It follows from this concept that when we think we have acase of different causes leading to the same effect, weproceed to make finer analyses (either spatial or sequential)in order to find what is common to the apparently differentcauses.

`Thus, multiple causation is a complication of enquiry at twolevels. First, it involves the simple possibility that an eventmay have "several causes." Second, it involves the moreabstruse notion that several causes may really be onethough,of course, the common factor, the "true" cause may belocated in several places or react in several differentcircumstances, or be involved in several different sequences.

`In this Invitation we deal with "multiple causation" in itssimplest form: thr idea of the same causal source lying inseveral different locations. We do not introduce thecomplication of an underlying common cause acting indifferent circumstances or through different sequences.' (p117)

INVITATION #24

Subject: Control of Thyroid Secretion (p 128)Topic: Inhibiting Causes

Invitations 17 and 21 have treated causal factors as beingpositive, leading to the appearance or the increase ofsomething. This Invitation illustrates the fact that a causalfactor may be negative and lead to the inhibition ofsomething.

INVITATION #25

Subject: PituitaryGonad MechanismsTopic: Feedback Mechanisms

`This Invitation has two major points. First, it exemplifies thefact that experimental tests of hypotheses cannot, as a rule,verify them. As a rule we can demonstrate only that thehypothesis is not possible or that it might be the case. Onlywhen all the possible alternative hypotheses are known canexperimental test, by eliminating all but one alternative,verify that one. This situation is sometimes referred to as

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CaloricContentof Food

"the falsifiability, but not the verifiability of hypotheses."The point is that we are rarely in a position to say withcertainty that we know all the possible alternatives.

`The second function of the Invitation is to add a last item toour understanding of cause-effect enquiry: the existence ofcomplex 'interactions of causes.

`The Invitation is based on a real casethe sexual cycle in themammalian female. However, it refers to organs and materialsonly by code letters, A, B, C, and so on. This is done for tworeasons. First, it makes it possible to use the Invitation at anytime, since background information is not required. Second,it permits us to put our emphasis on the general pattern ofinteraction, rather than on the specific case.' (p 130)

The previous Invitations are examples of how students caninvestigate the:

function of various glands,

relationship among gland organs and the hormonesproduced,feedback mechanism as a regulator of hormone secretion.

They stress the role of critical observation and involvestudents in designing experiments, interpreting data,formulating hypotheses, identifying problems, andperforming other tasks related to the investigative nature ofscience.

Students may determine the relative caloric content of foodby using a standard calorimeter or the homemade one shownin the sketch.

Thermometer (Celsius)(Do not leave in tubewhile heating.)

Soft drink can

Test tube (bottomshould be approximately2 cm above food.

Food

Needle

Cork

Calorimeter Cover cork withaluminum foil

Diagram from BSCS Patterns and Processes 1966 p. 7881

Heart andBreathingRates

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When using a homemade calorimeter, satisfactory results canbe obtained by following these procedures:

1) Measure 2/10 gram of food to be tested and place on theneedle;

2) Place 10 ml of water in the test tube and record thetemperature of the the water;

3) After the initial reading, remove the thermometer fromthe test tube;

4) Ignite the food and place calorimeter over burning foodwith flame directly under the test tube. Burn food to anash. If flame goes out, repeat steps 1, 2, 3, and 4.

5) Immediately measure the resulting increase of watertemperature;

Note: If water should boil, the amount of water shouldbe increased in multiples of ten; or decrease the amountof food. If the water boils, most of the heat energy isbeing used as heat of vaporization of the water (540calories per gram);

6) The caloric content of the food is calculated as follows:change in temperature (Celsius) x number of ml of water= calories present in the amount of food burned.

Note: This rocedure provides a caloric content in smallcalories (c). (c is the heat necessary to raise thetemperature of 1 ml of water 1°C.) There is a largerunit, the kilocalorie or calorie with a capital C, which is1000 times as big as the small c calories and is the unitcommonly used today in specifying caloric content offood.

Foods that provide particularly good results include nuts,cereals, sugar, and dried bread.

Students working in pairs can gather data on heart rate andbreathing rate using amount of activity as a variable. Onestudent can record the pulse rate and breathing rate ofanother student lying at rest, standing, after mild activity,and after more strenuous activity. The data gathered can betabulated in the following form.

Heart Rate Breathing RateLyingdownStandingat restMildactivityStrenousactivity

Class results can he graphed on each activity to illustrateindividual differences within the class. Average results for allactivities can be graphed to illustrate the effects of theamount of activity on these two body functions.

The heart of a double pithed frog can be attached to akymograph for observation of amplitude and rate of heartbeat. (For pithing instructions, cee Follansbee Harper AnimalBehavior, BSCS Laboratory Block, D. C. Heath & Company,p 27)

Laboratory stand

Thread attachedto frog heart

Kymograph

The heart can be cooled using iced Ringer's solution (asolution of salts comparable in composition to the frog'sbody fluid) to slow it down and a chemical, such as adrenalin(1 in 10,000), to speed up heart action.

Physical stimulation, i.e., poking with a pin or small electricalstimulation, will also alter the rate.

Lung Capacity To study the relative lung capacities, the setup shown in thesketch can be used. Have students, using a normal breath,blow into the rubber tubing and measure the amount ofwater displaced.

After refilling the jar a second student may repeat theexperiment. For comparative purposes students may wish torepeat the experiment after lungs to capacity andexhaling all the air in the lungs to displace the water.

For more accurate volume measurements, mark the waterlevel with a grease pencil, remove the jar, empty itcompletely, and with jar in upright position measure theamount of water necessary to fill the jar to the pencil mark.

Kymograph -an instrument forrecording variations or undula-tions, arterial or other

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CarbonDioxide under different levels of physical activity.Related toPhysicalActivity

Open mouth jar

0Rubber orplastic tubing

Tray

This experiment demonstrates the amount of CO2 produced

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Measure 100 ml of water and test for CO2 by adding 5 dropsof a 1% solution of phenolphthalein. If a pink color appearsin the water and remains for at least fifteen seconds, it can beassumed there is no acid in the water. If the water remainsclear, there is acid present and it must be neutralized beforethe amount of CO2 added can be determined. To neutralizethe water add dilute sodium hydroxide (NaOH) solution dropby drop until the water turns slightly pink. Be sure to countand record the number of drops required. Save the sample ofwater and use it as a control reference.

1) Have a student lie on his back inhaling through the noseand exhaling through a tube which is inserted in a flaskwith 100 ml of water containing 5 drops ofphenolphthalein for one minute.

2) Add sodium hydroxide (NaOH) drop by drop as beforeuntil the pink color appears as in the control solution.The difference in the number of drops of NaOH requiredto color the control solution and experimental solution isan indication of the amount of CO2 added.

3) For comparison purposes, the experiment can berepeated after exercising vigorously for one minute, e.g.,deep hand bends, running in place. etc.

4) Additional activity: have the student undergo somephysical activity such as running up a flight of stairs. (Thework required to run up the stairs can be calculated if sodesired.) Then have the student breathe into the

VisualIllusions

VestibularApparatus

PhotoperiodEffects

phenolphthalein solution for the same length of time asbefore. Note the number of drops of NaOH added toindicate a change.

If phenolphthalein is not available, the same relative meas-sures may be obtained by blowing into limewater and com-paring the amount of precipitate formed when CO2 reactswith the calcium hydroxide to form calcium carbonate.

Students may want to investigate some of the visual illusionsthat are discussed in most high school psychology textbooks.

Other sense investigations can also be used, i.e., have studentsmake a 1-inch grid of washable ink on the palm of the hand,back of the hand, and on the forearm. The grid should bemade up of 1/8-inch squares. By using 3 inch nails, some ofwhich have been placed in ice water and some in hot water.the students can attemp. to map the heat and cold receptorsin the grids. Note: one student should place the nails foranother student who is blindfolded. Students must take carenot to push too hard on the nails or the pain or pressurereceptors will be stimulated, confusing the experimentaloutcome.

By comparing the location and number of receptors in thethree grids, students can get an indication of the moresensitive areas to heat and cold.

The student may perform experiments that provide ameasure of the role and function of the vestibular apparatusin maintaining balance and posture. In order to stress thisbody system, the student can be seated in a swivel chair andspun "X" number of times. The effect of this stress can bequalitatively observed by assignment of selected tasks such aswalking a straight line, picking up small objects, tossing coinsinto a small bucket, etc.

English sparrows may be captured during the winter monthswhen the photoperiod is approximately nine hours. The birdsmust be sacrificed in order that the gonads may be examined.(A painless way to kill the birds is to dip their beaks into thefumes of carbon tetrachloride. Hold the birds beak into theopening of a bottle of CCL4. Students should avoid breathingthe fumes.)

Measure the weight and size of the gonads of a male andfemale. This serves as a control.

The remaining birds are to be placed in covered cages with alight and timing device to increase the photoperiod to 13 or14 hours. Birds should be exposed to the lengthenedphotoperiod for three weeks. One pair should be kept underthe 9-hour photoperiod to serve as additional control.

8b

CircadianRhythms

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At the end of the three-week period, the size of the gonads ofthe control and experimental birds may be examined.

Circadian periods such as photoperiodicity can beinvestigated in the laboratory or classroom. Mice can beplaced in a cage with an activity wheel. A 24-hour recordingdevice attached to the wheel can be used to record thenumber of revolutions and time of day. By using timers tocontrol artificial lighting, the photoperiod can be set to covera wide range of light-dark cycles. Analysis of activitypatterns, light-dark cycles, food consumption, and bodyweight will reveal the influence of photoperiod.

Section 9

Glossary of Terms

GLOSSARY OF TERMS

Aberration

Blastoid

Biorhythm

Any deviation from the normal number.

Pertaining to embryonic cell forms

A biologically inherent cyclic variation or recurrence of an event or state,such as the sleep cycle, circadian rhythms.

Cation An ion carrying a charge of positive electricity; therefore going to thenegatively charged cathode.

Chromosomal Pertaining to the body in the cell nucleus that is the bearLr of genes.

Circadian Relating to biologic variations or rhythms with a cycle of about 24 hours.

Colchicine An alkaloid obtained from colchicum.

Cytogenetics Study of hereditycytology and genetics.

Diastolic Relating to the dilation of the heart cavities, during which they fill withblood.

Diuresis Increased discharge of urine.

Diurnal Pertaining to day or daylight.

Endocrine The internal secretion of a gland.

Endogenous Originating or produced within the organism or one of its parts.

Epinephrine The chief hormone of the normal adrenal medulla, adrenaline.

Erythropoiesis The formation of red blood cells.

Exogenous Originating or produced outside.

Friability Easily crumbled or reduced to powder.

Globulin A simple protein found in blood, milk, muscle and seeds.

Hematopoietic Blood forming.

Hemolysis The alteration or destruction of red blood cells in such a manner thathemoglobin is liberated into the medium in which the cells are suspended.

Hemostatic Arresting the flow of blood within the vessels.

Histochemical Chemistry of the tissues.

Homeostatic The state of equilibrium in the living body with respect to variousfunctions and to the chemical composition of fluids and tissues.

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Homeothermic Denoting the warm blooded animals.

Humoral Relating to the extracellular fluids of the body.

Hyperbaric Pertaining to pressure of ambient gases greater than 1 atmosphere.

Hypobaric Characterized by low atmospheric pressure.

Hypodynamia Diminished power.

Hypotension Subnormal arterial blood pressure.

Leukocyte Any one of the white blood cells.

Lysis Destruction, as of cells by a specific substance (lysin),

Mesenchymal A pluripotential cell form found between the ectoderm and entoderm ofyoung embryos. These cells give rise to any of the connective orsupporting tissue.

Microflora Microscopic forms of bacterial life.

Mitosis The usual process of cell reproduction consisting of a sequence ofmodifications of the nucleus that result in the formation of two daughtercells with exactly the same chromosome and deoxyribonucleic acid(DNA) content as that of the original cell.

Morphologic Relating to the science which treats of the configuration or the structureof animals and plants.

Myocardium Heart muscle.

Neutrophil A mature white blood cell in the granulocytic series.

Oculogyria The limits of rotation of the eyeballs.

Orthostatic Relating to or caused by the erect posture.

Organelle One of the specialized part of a protozoan or tissue cell serving for theperformance of some individual function.

Osteoblastic Relating to a bone forming cell.

Osteoclastic Relating to the activity in the absorption and removal of osseous (bony)tissue.

Otolith Part of the vestibular apparatus (ear stone).

Parenchymal The distinguishing tissue of a gland.

Pathologic Diseased.

Phagocytosis Engulfing of microorganisms, other cells, and foreign particles byphagocytes.

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Poikilothermic Denoting the so-called cold blooded animals and the plants.

Reticulocyte A young red blood cell.

Spatial Relating to space, or a space.

Steroid Hormone of the adrenal cortex.

Stressor Any force which stresses the body, organ, or system.

Systolic Relating to the rhythmical contraction of the heart.

Venous Relating to a vein or to the veins.

o U. S. GOVERNMENT PRINTING OFFICE : 1973 724-916/9,1

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