MANUAL FOR

PHYSIOLOGY LAB WORK

Slides made and arranged by:

Prof. dr Jelena Popadić Gaćeša

in cooperation with:

Ass. dr Vedrana Karan

Ass. dr Danijel Slavić

and lab technicians

Marija Bjeljac

Dejana Ravnjak

Bruno Krčmar

EXCITABLE TISSUES

Equipment for excitable tissues’ lab work

Electrodes without polarization (Zn + ZnSO4)

Device for stimulation with direct current (UDJS)

Equipment for excitable tissues’ lab work

Electrical stimulator

Equipment for excitable tissues’ lab work

Kymograph

Equipment for excitable tissues’ lab work

Preparations in neurophysiology

galvanoskopic leg sample

suspension method vertically and horizontally

neuromuscular sample

Transport through membrane

pasive transport

Na+

O2

fructose

transporter

simple diffusion transport

through ionic

channels

facilitated diffusion

active transport

vesicular transport

endocytosis exocytosis

primary active

transportsecondary active

transport

Na+ Na+Na+

ATP ADP+Pi

K+ K+Na+

glukoza

Transport through membrane Electrical potentials of the membrane

resting

membrane

potential

outside

inside

Action potential (AP)

membrane potential (mV)

overshoot

threshold

hyperpolarization

time (ms)

Nerve fibers division

velocity (m/s) diametar (μm) type functions

70–120 12–20 Aα motor neuron, inervates skeletal muscle

30–70 5–12 Aβ fine touch, vibration

15–30 3–6 Aγ motor neuron, inervates muscle spindles

12–30 2–5 Aδ sharp pain, cold

3–15 < 3 B vegetative nervous system

0,5–2 0,3–1,2 C dull pain, warm, itch, tickle, sexual sensations

Action potential conduction

-in myelinated fibers

- saltatory conduction

- fiber diametar 1-20 μm

- conduction velocity 6-120 m/s

-in unmylinated fibers

- continuous conduction

- fiber diametar about 1μm

- conduction velocity1 m/s

Ranvier node myelin sheet

neuron

Du Bois-Reymond law

stimulus response

1. intensity

2. duration

3. slope

Relationship between stimulus intensity

and duration of stimulation

signal intensity (I)

chronaxie (C)

time

time (t)

rheobase (R)

Polarization current

+ -direct current

--

--

-

-- - -

++

++

+

++ + +

polarization current

Electrotonus

+ -konstantna struja

hypopolarization =

katelectrotonus

+-- +

+ + ++ ++ +----- - -- - - - -

+- - --- - -

+ -direct current

+-- +

+ ++++ + ++++++++ +----- - -- - - - -

+- - --- - -

MRP

Electrotonus

+- -- - - - -

+

+ -direct current

--

hyperpolarization

= anelectrotonus+ ++++ + ++++++++ +

- ------- +- - -

MRP

MUSCLES

Types of muscles in human body:

• striated

• smooth • heart

Skeletal muscle

blood vessel wall

heart

muscle

muscle fiber

nuclei

striated striated

muscle fiber

nucleus

discus

intercalates

muscle fiber

nucleus

Specificities - structure

characteristics skeletal smooth cardiac

distribution skeleton internal organs heart

hystology striated smooth striated

contractile elements actin and myosin actin and myosin actin and myosin

distribution of miofibrils A:M = 2:1 irregular A:M = 2:1

number of nucleus ~ 100 1 1

sarcomere + - +

sarcoplasmatic reticulum ++ - +

T-tubules triade - diade

Ca++ sarcopl.reticulum extracellular sarcopl.reticulum

syntitium - +/- +

characteristics skeletal smooth cardiac

elasticity + +/- +

plasticity - + -

hierarchy - - +

automatism - -/+ +

characteristics of contraction

fast contraction and decontraction

slow contraction and decontraction

prolonged contraction

energy sources glucose, FFA glucose, FFA lactate, FFA, glucose

action potentialdepolariz.(Na+)/

repolariz.(K+)1-2 ms

depolariz.(Na+/Ca++)/repolariz.(K+)

10-20 ms

plateau (Ca++)

300 ms

Specificities - function

characteristics skeletal smooth cardiac

innervation alfa motoneuron VNS (Sy and PSy) automatismVNS (Sy and PSy)

movements voluntary involuntary involuntary

motor plate + - -

neurotransmitter AcCh Epi, AcCh Epi

receptors nicotinic gap junctions gap junctions

motor unit + - -

gap junctions - +/- +

Specificities - innervation

Action potential of cardiomyocyte

plateau

repolarizationdepolarization

Single-twitch muscle contraction =

myographAB – latent period

BC – contraction phase

CD – decontraction phase

A B C D kymographic recording

Neuromuscular junction = motor plate

axon

action

potential

AcCh-esterase

presynaptic membrane synaptic

cleft

postsynaptic membrane -

sarcolemma

AcCh

Influence of t on single-twitch muscle

contraction

t

LP, CP, DP

amplitude

t

LT, CP, DP

amplitude

t

t

kymographic recording

contraction

phase (CP)

decontraction

phase (DP)

late

nt

per

iod

(L

P)

Motor unit

alpha-motor neuron

+

muscle fibers

“all or none” response

motor plate

axon terminal

Influence of signal intensity on size of

muscle contraction

kymographic recording

Division of muscle contractions

- impulse number

single-twitch and tetanic

- length or tone lenght

isotonic and isometric

- movement

static and dynamic

- shortening or lenghtening

concentric and eccentric

Tetanic muscle contraction

Z – incomplete tetanus

G – complete tetanus

f

in decontraction phase in contraction phase

summation

Sources of E for contraction

• aerobic

– CH

– fats

• anaerobic

o ATP

o ADP

o CP

o anaerobic glycolysis

Sources of E for contraction

– carbohydrates

Krebs cycle

and

oxidative

phosphorylation

ATP

glucose pyruvate

2

lactate

O2

anaerobic glycolysis

ATP36Σ transport of e- across mitochondrion

membrane–

mitochondrioncytoplasm

AcCoA

O2

Sources of E for contraction

– fats

Krebs cycle

and

oxidative

phosphorilation

beta oxidation

ATPΣ depends on C atom number

AcCoA

O2

Glycerol

+

3 fatty acids Acil-CoA +

HOOC – CH2 – CH2 – ... – CH2 – CH2 – CH3 + CoA fatty acid

mitochondrion

Work and power

J = N m

A = F s

force

distance

P = A

t

work

time

W = J/s Muscle work is the greatest with medium loads.

Power of muscle contraction depends on: - physiological CSA

- resting fiber length

- impulse frequency and intensity

- muscle origins and insertions angles

- angles between joints etc.

Mechanical efficiency (ME)

ME = 2-30%

chemical E

ATP

mechanical E = work

thermal E = heat

hypertrophy

hyperplasia atrophy

muscle

CSA/Vol

muscle

CSA/Vol

CSA/Vol of myocyte

( n of filaments)

n of myocytes

CSA/Vol of myocyte

( n of filaments)

Muscle fatigue

causes of fatigue

centralperipheral

Muscle sorreness

micro traumas of muscle fibers

Signs of inflammation: pain

redness

temperature

edema

function

RESPIRATION

Respiration

• ventilation

• gas exchange between alveoli and blood

• gas exchange between blood and intercellular fluid

• cell breathing

Mechanics of respiratory movements

• inspiratory musculature– diafragm

– mm.intercostales externi

– additional musculature(m.sternocleidomastoideus,

mm.serrati ant.,

mm.scaleni)

• expiratory musculature

–mm.intercostales interni

– abdominal musclesmm.intercostales

externi

mm.intercostales

interni

diafragm

Interpleural pressure

parietal pleura

visceral pleura

interpleural

space

interpleural pressure

-8 to -1 mmHg

Donders model

+

0

-

+

0

-

1.

rest

-4 mmHg

Donders model

+

0

-

+

0

-

2.

inspirium

- 8mmHg

Donders model

+

0

-

+

0

-

3.

expirium

Donders model

+

0

-

+

0

-

4.

Müller manoeuvre(inspirium with closed airways)

Donders model

+

0

-

+

0

-

5.

Vansalva manoeuvre(expirium with closed airways)

Donders model

+

0

-

6.

pneumothorax

+

0

-

Surfactant

• Alveolocites type II

• from 7. intrauterine month

• Functions: • decrease of surface tension in alveoli

• allows equal filling of alveoli

• prevents collaps of alveoli

Disturbance in secretion – respiratory distress syndrome (RDS)

neonatal adult

Respiratory unit

Alveolus + capillary

Ventilation disturbance – lung bases

V/Q ratio ≈ 1

V => V/Q tends to 0 Q => V/Q teži ∞

Perfusion disturbance – lungs apexes

Q – perfusion (vascularisation)V – ventilation

Gas diffusion through respiratory

membrane

Diffusion flux depends on:

– diffusion coeficient (gas solubility)

– differences in partial pressure (pO2, pCO2)

– membrane surface

– membrane thickness

– temperature

same for O2 and CO2

Pneumography

• kinds of breathing

• types of breathing

• thoracal

• abdominal

• mixed

Vrsta disanja Vt (ml) f (udaha/min)

Eupnea 500 12-16/min

Tachipnea N

Bradipnea N

Hypopnea N

Hyperpnea N

Polypnea

Oligopnea

Apnea - -

Dyspnea – subjective feeling of air lacking

Ortopnea – dyspnea that depends on body position

Spirometry

Vt

IRV

ERV

RV

TLCVC

IC

FRC

Lung volumes:

Vt – 0.5l

IRV – 2.5-3l

ERV – 2-2.5l

RV – 1.2l

Lung capacities:

VC – 3-7l

TLC – 4-8l

IC – 3-3.5l

FRC – 3-3.5l

Forced spirography

FEV1%

N FEV1%

N FEV1%

FEV1%=

FEV1

FVC

FEV1%- opstructive ventilatory disorder

asthma

chron.bronchitis

emphysema

N FEV1%- restrictive ventilatory disorder

pneumonia

atelectasis

pneumothorax

lung edema

lung fibrosis, ...

Transport of gases through blood

• physically dissolved

• chemically bonded

O2 CO2

3%

97%

Physically dissolved

Chemically bonded:

oxyhemoglobin

O2-Hb

Physically dissolved 10%

90%

Chemically bonded:

bicarbonate HCO3-

carbonic acid H2CO3

carbaminohemoglobin

CO2-Hb

Composition of inhaled, exhaled and

alveolar air

inhaled air

O2 – 20.8%

CO2 – 0.04%

exhaled air(inhaled + alveolar)

O2 – 15.7%CO2 – 3.6%

alveolar air

O2 – 13.6%

CO2 – 5.3%

Regulation of breathing

• Neural– voluntary

– center for breathing

– reflex

• Humoral• O2

• CO2• pH

BLOOD

TRANSPORT

gases – O2 i CO2

nutrients

metabolic waste products

TRANSPORT OF HORMONES

IMUNOLOGICAL

HEMOSTASIS

TERMOREGULATION

MAINTAINANCE OF A-B BALANCE – BUFFERS

FUNCTIONS OF BLOOD

BUFFERS

BUFFERS IN BLOOD PHYSIOLOGICAL BUFFERS

ORGANIC:

Hb and plasma proteins

NEORGANIC:

bicarbonate, phosphate

LUNGS

KIDNEYS

HOMEOSTASIS

constant blood composition

BLOOD pH 7,35 – 7,45

Acid-base balance

• pH 7.35 – 7.45

• PaCO2 – 40 mmHg

• HCO3- – 24 mmol/l

Physiological compensatory

mechanisms

• Blood buffers

• Lungs – ventilatory changes

• Kidneys – H+ excretion and HCO3- synthesis

physiological

buffers

Blood buffers

• Blood buffers

– hemoglobin

• Plasma buffers

organic inorganic

- plasma proteins

- albumins

- globulins

- fibrinogen

- bicarbonate

- phosphate

- amoniac

BE (base excess) 20:1 –20x more base components in blood buffers

Physiological buffers

• Lungs

– alveolar ventilation

• Kidneys

– regulation of HCO3- plasma concentration

– excretion of acidic metabolites (NH4+, H2PO4

-)

Acid-base balance disorders

• ACIDOSIS

– metabolic

– respiratory

• ALCALOSIS

– metabolic

– respiratory

HCO3-

pCO2

HCO3-

pCO2

BLOOD

PLASMA CELLULAR ELEMENTS

55% 45%WATER 90%

HEMATOCRIT ELECTROLITES

PROTEINSI

60-80 g/l

ALBUMINES

GLOBULINES

FIBRINOGEN

α1 i α2

ß

γ = Imunoglobulines Ig:

7% = 5-6l

IgG, IgM, IgA, IgE, IgD

Hct 0.40 – 0.45

1h

2h

3-10 mm

8-16 mm

SE

SEDIMENTATION

-Number

-Shape and size

-Creation/degradation

-Life span

-Cellular characteristics

-Functions

-Disorders

ERYTHROCYTE NUMBER COUNT

3mm

1mm

1/40mm1/20mm1/5mm

V = 1

20

1

20

1

10mm3

V = 1

4000mm3

ERYTHROCYTE COUNT

Br Er =N

n4000 R 106

n = 16 5 = 80

N – Er number

n – square number

4000 – correction (to 1mm3)

R – dilution (100 or 200)

106 – in l (dm3)

V = 1

4000mm3

HEMOGLOBIN

Concentration 120 – 180 g/l

Function – gases transport

Types of Hb

Hb + O2 = OXY-HEMOGLOBIN

Hb + CO2 = CARBAMINO-HEMOGLOBIN

Hb + CO = CARBOXY-HEMOGLOBIN

Hb – O2 = DEOXY-HEMOGLOBIN

Hb-Fe3+ = MET-HEMOGLOBIN

globine

globine hem

Fe2+

bilirubine

protoporphyrine

Fe2+

IRON – Fe

TRANSFERRIN

FERRITIN

HEMOSIDERIN

transport

depot

BLOOD

LIVER

Daily intake – 5 mg, apsorption 10%

Sources of Fe

ERYTHROCYTE INDEXES

MCV =

MCH =

MCHC =

conc. Hb (g/l)

nr. Er/l blood≈ 30 pg

Hct

nr. Er/l blood

≈ 90 fl

conc. Hb (g/l)

Hct≈ 300 g/l

HEMOLYSIS

physical

chemical

biological

imunological

OSMOTIC RESISTANCE OF

ERYTHROCYTES

Hypotonic solution hemolysis

Minimal osmotic resistance 0.42-0.46% NaCl

Maximal osmotic resistance 0.30-0.34% Na Cl

Range of osmotic resistance 0.30-0.46% NaCl

DECREASED CREATION

OF Er

INCREASED

DEGRADATION OF

Er

BLEEDING

TypeType::Aplastic

Megaloblastic

Sideropenic

Hemolytic

CauseCause::

-B12

- folic acid

Fe

Loss of erythrocytes

ANEMIAS

LEUKOCYTE COUNT

Nr Leu =N

n10 R 106

n = 1 4 = 4

N – Leu number

n – square number

10 – correction (to 1mm3)

R – dilution (10 or 20)

106 – in l (dm3)

V = 1

10mm3

V = 1 1 1

10mm3

DIFFERENTIAL BLOOD COUNT

Neutrophils 50-75%

Eosinophils 2-4%

Basophils 0-1%

Lymphocytes 20-40%

Monocytes 2-10%

Granulocytes (polymorphonuclears)

Agranulocytes (mononuclears)

BLOOD SMEAR

THROMBOSISBLEEDING

HEMOSTASIS

1. MAINTAINANCE OF

BLOOD IN THE BLOOD

STREAM2. PREVENTION OF BLOOD

COAGULATION IN THE BLOOD

STREAM

3. STOP OF BLEEDING AFTER INJURY

ProcoagulantsAnticoagulants

HEMOSTASIS

1. VASOCONSTRICTION

2. CREATION OF

PLATELETS CLOT

3. COAGULATION – FACTORS OF COAGULATION

4. FIBRINOLYSIS

BLEEDING TIME

- Ivy (6-7 min.)

- Duke (1-3 min.)

COAGULATION TIME

- Foney (10-18 min.)

BLOOD TYPES

blood type A B AB 0

agglutinogen A B A, B -

agglutinin anti-B anti-A -anti-A,

anti-B

agglutination

Blood derivates

• blood

• fresh frozen plasma

• erythrocytes

• platelets

• ...

BLOOD TYPING

anti-A anti-B anti-Rh

serums

KIDNEYS

Urine formation

• Glomerular filtration

– Primary urine – plasma ultrafiltrate

– GFR (glomerular filtration rate)

= 125ml/min or 180l/day

- Neto filtration pressure – 10mmHg

• Tubular reabsorption and secretion

– sekundary urine

– Diuresis 1.5 – 2l/day

Blood Plasma Primary urine Final urine

Cellular elements + - - -

Water + + + +

Salts + + + +

Proteins + + - -

Amino acids + + + -

Glucose + + + -

Waste metabolic

products

+ + + +

Harmful substances + + + +

glomerul

Primary urine

Filtration pressurehydrostatic pressure

in capillaries

oncotic pressure

in capillaries

hydrostatic pressure of

Bowman capsule–= –

10mmHg 60mmHg 32mmHg 18mmHg= ––

a.afferens a.efferens

filtration of plasma – 125ml/min

Proteins are not filtrated because:

- molecule size

- electrical charge

(“anion”)

proximal

tubule

distal tubule

Henle’s loop

thin descendent

segment

thin ascendent

segment

thick ascendent

segment

collecting

duct

primary

urine

final urine

H2O (65%)

Na+ (65%)

Cl- (65%)

HCO3- (65%)

K+ (70%)

Ca++

glucose

amino acids

Mg++ (25%)

H+

Na+ (25%)

Cl- (25%)

K+ (25%)

Mg++ (65%)

Na+ (4-7%)

Cl-

Mg++ (1-5%)

Na+ (do 2%)

Cl-

H+

K+

distal tubule

collecting ducts –

hormonal dependent

reabsorption:

ADH

Aldosterone

Parathormone

H2O (20%)

H2O

Urine characteristics

• amount

• appearance

• color

• smell

• specific gravity

• pH

• osmolarity

• diuresis oko 1.5 l

• clear

• yellow

• aromatic

• 1.015-1.022 (1.003-1.040)

• 5-6 (4.8-8)

• 500-800mosmol/l

Urine characteristics - amount

Diuresis

1.5l/dan

anuria

< 150ml/dan

oliguria

150-500ml/dan

polyuria

> 2l/dan

Urine characteristics - appearance

opaque

becomes transparentheating or reaction

with base

uratesphosphates

adding acetic acid

adding hydrochlorid

acid

oxalates

Opacity remains:

-Er

-Leu

-Epithelial cells

-Bacterias

-Salts

-Fats

-Pus

-Mucus

Urine characteristics – color

• Urobilinogen - 1

Urine color change:

-food (e.g. beet)

-medications

-Er

Hematuria

Micro-

some Er, urine color

remains unchanged

Macro-

red color of urine

Urine characteristics – specific gravity

Hyposthenuria – damaged kidney urine-concentrating ability

Isosthenuria – no concentration – specific gravity of primary and finalurine is the same

Hypersthenuria – dehydration or presence of pathological elements in urine

In single urine portions specific gravity in range

1.003-1.040

Urine characteristics – pH

pH 5-6 (4.8-8)

Plant food,

vegetariansAnimal food

Chemical composition of urine

• Qualitative analysis of chlorides

urin

AgNO3

White precipitate

of AgCl

Chemical composition of urine

• Quantitative analysis of chlorides

AgNO3 + NaCl AgCl + NaNO3

Urine

+

K-chromate

AgNO3

Beli talog AgCl – dok ima hlorida

white precipitate

red precipitate Ag-chromates

(consumable AgNO3 – 1ml) 0.01 150 = ml/day NaCl

Chemical composition of urine

• Qualitative analysis of sulphates

urin

BaCl2

Beli talog BaSO4

neorganski sulfati

Ba(OH)2

krupni talog BaSO4

ccHCl

filtrat

heat

white precipitate BaSO4

organic sulphates

Chemical composition of urine

• Qualitative analysis indicane

urine

ccHCl FeCl3

indigo blue

Proteinuria in physiological conditions

• Probe with cc HNO3

urine

ccHNO3

Heller’s

ring

+ reaction

-physical work

-swimming in a cold water

-pregnancy • Probe with sulfosalicylic acid

Proteinuria in physiological conditions

urine

sulfosalicylic

acid

opacity

reaction+

— reaction no opacity

white opaque

+ barely visible opacity – proteins in traces

++ visible opacity – clearly positive result

+++ creation of small flakes – strongly positive

++++ coarse sediment – very positive

• Probe with heating

Proteinuria in physiological conditions

heating

urine

acetic

acid

opaque

reaction+

— reaction opacity disappears by adding acetic

acid (precipitate from carbonates or

phosphates)

opacity remains after adding acetic

acid

Glycosuria in physiological conditions

• Fehling’s probe

Fehling I

+

Fehling II

urine

heat

red precipitate

-per os intake od large amounts of monosaccharides

-iv. infusion of monosaccharides

Ketonuria in physiological conditions

• Legal’s probe (probe with sodium nitroprusside)

urine

Na-nitroprusside NaOH

-starvation

-pregnancy

-acetone

-acetoacetic acid

-Beta oxybutyric acid

Ketonuria in physiological conditions

• Lieben’s probe (iodoform probe)

urine

Lugol’s solution

(aqueous iodine solution)NaOH

iodoform

Urine sediment

• Salts

– Urates

– Oxalates (Ca-oxalate)

– Phosphates (ammonium, Ca, Mg-phosphate)

• Epithelial cells

• Erythrocytes – up to 5 cells

• Leukocites – up to 10 cells

Plasma clirens

• Amount of plasma cleared of some substance in

the unit of time

– Creatinine – 1 (≈125ml/min)

– Proteins – 0 (no filtration)

– Glucose – 0 (complete reabsorption)

ENDOCRINOLOGY

Insulin

• Insulin synthesis

• Insulin secretion

• Insulin acts on following organs:– liver

– skeletal muscle

– adipose tissue

Leading

segmentB-chain C-peptide A-chain

preproinsulin

B-chain C-peptide A-chain

proinsulin

B-chain

A-chain

insulin

--

-S

S

S

S--

-

C-peptide

Metabolic functions of insulin:

• Carbohydrates– utilisation of glucose from circulation ( BG)

– glycolysis

– glycogenesis

• Fats– synthesis of free fatty acids

– depot of fats

• Proteins– synthesis of proteins

– depot of proteins

– glyconeogenesis

in circulation

glucose

fatty acids

amino acids

Glycemic curve in OGT test

120 min60 min 180 min0 min time

Glycemia (mmol/l)

5

10

15

20

GLUCOSE INTOLERANCE

PHYSIOLOGICAL AREA

Oral glucose tolerance test (OGT) in

physiological conditions

OGT TEST Glycemia (mmol/l)

measurement time referral values pathological values

0 min 3.3 – 5.5 mmol/l > 7.2 mmol/l

60 min < 8.88 mmol/l > 12.2 mmol/l

120 min < 6.6 mmol/l > 7.77 mmol/l

180 min below 0 min > 7.2 mmol/l

DIGESTION

DIGESTIVE TRACT

• DEFINITION

• FUNCTIONS:

– INGESTION (food intake, cheewing, swallowing)

– DIGESTION (mechanical, chemical)

– ABSORPTION (passive, active)

– AMINO ACIDS, CHOLESTEROL, GLUCOSE, VITAMINS, MINERALS, WATER – DIRECT ABSORTION

CHEMICAL BRAKE DOWN OF FOOD

– CH – monosaccharides

- FATS – free fatty acids

- PROTEINS – amino acids

• CONDITIONS FOR DIGESTION

– PRESENCE OF SUBSTRATE

– PRESENCE OF ENZYME

– TEMPERATURE

– OPTIMAL pH

– TIME

DIGESTION IN ORAL CAVITY

• CARBOHYDRATES• Starches + saliva (alpha-amylase) disaccharides

– Fehling probe red precipitate

• PROTEINS• Egg white + saliva no reaction

– Biuret reaction solution remains blue

• FATS• Oil + saliva no reaction

– Probe with chrome-sulfuric acid solution remains orange

t

t

t

+

-

-

Fehling reaction

Fehling I: CuSO4

Fehling II: C4H4KNaO6 + NaOH

CH2OH(CHOH)4CHO + 2CuO CH2OH(CHOH)4COOH + Cu2O

red precipitate

Fehling I

+

Fehling II

supernatant

with saliva

starches Fehling I

+

Fehling II

heat

• Biuret probe

– Reagents: NaOH + CuSO4

(H2N-CO-)2NH

biuret

Cu2+

O C

H N

C R

O C

H N

C R

C O

N H

R C

C O

N H

R C

+ NaOH

+ CuSO4

rastvor ostaje

plave boje

positive

reaction -

solution

becomes velvet

Supernatant

with saliva

Egg white

supernatant

• Probe with chrome-sulfuric acid

K2Cr2O7 + H2SO4 + H2O 2H2CrO4 + K2SO4

orange solution green solution

+ K2Cr2O7

+ H2SO4

Supernatant

with saliva

oil

supernatant

positive

reaction -

solution

becomes green

due to glycerol

solution remains

orange

Rhodane probe in saliva

Saliva + FeCl3 + 6 NaCNS Na3Fe(CNS)6 + 3 NaCl

Saliva

red solutionexcrete

secrete

Digestion in stomack

• Gastric glands (types of gastric cells)

• Gastric juice composition

• Stomack motility

FUNCTIONS OF HCl

• pH

• bacteriostatic

• Denaturation of proteins

• activation of pepsinogen

• Fe3+ into Fe2+

REGULATION OF GASTRIC

SECRETION

• CEFALIC – neural

• GASTRIC – neural/humoral

• INTESTINAL – neural/humoral

• HUMORAL FACTORS: - GASTRIN

- AcCh

- HISTAMINE

- GIP

- SECRETIN

- CCK

Qualitative probe on gastric juice acidity

universal indicator – read the pH values

pH of gastric juice – 1-2

Concentration of HCl in gastric juice – 160mmol/l

Quantitative probe on gastric juice acidity

10 2 3 4 5 6 7 8 9 10 11 12 13 14

dimethylaminoazobenzol

phenolphtalein

gastric juice

Gastric juice

+dimethylaminoazobenzol

+phenolphtalein

NaOH

I

II

III

Referral values in 1000ml of gastric juice:

Total HCl = 500-600ml

Free HCl = I 300-400ml

Bonded HCl = Total HCl – free HCl 200-250ml

Total acidity = III 600-650ml

Acidity of other equivalents = III - 50ml

II+III2

II+III2

consumption in ml 0.00365 10 = g% HCl

Digestion in small intestine

small intestine juice composition

regulation of secretion

Pancreas

pancreatic juice composition

regulation of secretion (cephalic, gastric, intestinal phase)

Bile

bile composition

functions in digestion

regulation of secretion

THE SENSES

SOMATIC SENSATIONS

• Mechanical (fine touch, crude touch/pressure, vibrations, tickle and itch sensations, sexual

sensations)

• Thermal (heat, cold)

• Pain (mechanical, thermal, chemical stimuli)

• receptors

• nerve fibers

• pathways

Exteroreceptive sensations

(touch, heat)

• Fine touch

• Crude touch

• Heat

Proprioceptive sensations(pressure, vibrations, position,movement)

• vibrations

• kinesthesia

• myesthesia

• graphesthesia

• barognosia

THE EYE

• Light-refraction components

– Cornea

– Aqueous humor

– Lens

– Vitreous humor

• Light-reception components

– Retina

Refractive power of the eye

• Diopter

• Accomodation

Refractive power Retina Lens Eye

distant objects ~40 D ~20 D ~60 D

near objects ~40 D ~30 D ~70 D

Watching near and distant objects

accomodation

Formation of an image in complex

optical system of the eye

Image of near object which is formed by

refraction through the convex lens:

-realistic

-smaller

-upside down (inversed)

-reversed (as an image in a mirror)

Image of an object which is formed by

refraction through the concave lens:

-imaginary

-bigger

- straight up

Image of an object from infinitive distance

which is formed by refraction through the

convex lens is

point

Snellens charts

presbyopia

myopia

hyperopia

astigmatism

optotips

The Retina

RODS CONES

rod-like shape cone-like

125 milions number 5 milions

periferal localization central

shades of grey function color sight

periferal sight, smooth precisesness central sight, sharp

rhodopsin photochemical photopsin

slower, but better dark adaptation fast, but weaker

Color sight assessment

• Ishiharas charts

• 3 kinds of cones

• Types of color blindness

Visual pathway

Optic nerve n.opticusOptic

chiasm

Optic

tracttractus

opticus

Lateral

geniculate body

corpus

geniculatum

laterale

Optic

radiationradiatio

optica

Visual cortex

Brodmann 17, 18, 19

Perimetry

The Field of Vision

The Field of SightShape of the field of vision

Wideness of the field of vision

Pupillary Reflexes

reflex myosis mydriasis accomodation

receptors cones and rods

afferent nerv n.opticus

center Westphal-Edingers nucleus centrum ciliospinale Westphal-Edingers nucleus

efferent nerv n.oculomotorius Sy n. facialis

effector m.sphincter pupillae m.dilatator pupillaem.ciliaris

m.sphincter pupillae

Ophthalmoscopy

Fundus of the eye

-The yellow spot

-The blind spot

-Blood vessels

Mariotte’s experiment

• The blind spot– physiological scotoma

• Abnormalities in the Fields of Vision:

– Scotoma

– Hemianopsias (blindness in one half of the visual field)

• homonymous

• heteronymous

– Quadrantanopsias (blindness in a quarter of the visual

field)

Consecutive images

• Lateral inhibition

– Horizontal cells – for improving of contrast

– Bipolar cells

– Ganglion cells

• Excitation – excitated cell generates action potential

• Inhibition – surrounding cells are inhibited by the lateral

inhibition mechanism

• Under the change in light intensity, opposite phenomenon

occurs

THE EAR

• The external ear

• The middle ear

• The inner ear

• Frequency – level (Hz)

• Intensity – volume (dB)

• Bone conduction

• Air conduction

Tuning fork tests

• Rinne test

• Schwabach test

• Weber test

For an examination of:

air and bone conduction

lateralization of sound

Tone Audiometry

Tone audiogram

Examination of balance

• The Romberg’s test

– The test is performed in upright position with feet together. Arms are straight forward.

– Eyes are closed, and examiner stands close to the subject.

– The test is positive if the subject sways or falls.

The

Central

Nervous

System

The Spinal Cord

• Functions

– Reflexes

– Conduction

• Afferent pathways (tractus spinotalamicus and lemniscus

medialis)

• Efferent pathways (pyramidal and extrapyramidal)

Reflexes

• Definition

• Division of reflexes:

– Biological significance (alimentary, defensive, locomotor,...)

– Receptors (extero-, intero-, proprio-)

– Localization of reflex center (spinal, bulbar, pontile, cortical)

– Complexity of reflex center (mono- i polysinaptic)

– Clinical significance (depep, superficial, vegetative)

– Pavlov’s division (unconditional and conditional)

Reflex arch

1. Receptor

2. Afferent fiber

3. Center

4. Efferent fiber

5. Effector

Evaluation of different functions of

nervous systema on the spinal frog model

• Spinal shock

• Reflex time for flexion

• Spatial and temporal summation

• Iradiation of reflexes

• Withdrawal reflex

• Stepping reflex

• Periferal inhibition of reflex reaction

Clinically significant human reflexes

- muscle (deep) reflexes

reflex biceps triceps patellar Achilles

receptor for stretching

afferent nerve n. musculocutaneus n. radialis n. femoralis n. ischiadicus

center C4–C6 C6–C7 L2–L4 L5–S2

efferent nerve n. musculocutaneus n. radialis n. femoralis n. ischiadicus

effector m. biceps m. triceps m. quadriceps m. gastrocnemius

Clinically significant human reflexes

- superficial reflexes of skin

and mucous membrane

reflex abdominal plantar conjunctival

receptor for touch

afferent nerv segmental n. tibialis n. trigeminus

center Th8-Th12 L5-S2 medulla oblongata

efferent nerv segmental n. tibialis n. facialis

effector muscles of abdominal

wall

apropriate muscles m. orbicularis oculi

Reaction time

• Reflex time

• Reaction time

– Simple/complex

– Visual/acustic stimuli

Electrophysiological metods –

electroencephalography (EEG)

Electrophysiological metods–

electroencefalography (EEG)

Alpha rhythm – restfulness, closed eyes, f 8-13Hz, A up to 100 µV

Beta rhythm – f 14-30Hz, A up to 25 µV

Theta rhythm – f 4-7Hz, A up to 150 µV, superficial phases of non-REM, stress, kids

Delta rhythm – f 0.5-3Hz, A up to 300 µV, deep phases of non-REM

Gamma rhythm – f above 30Hz, learning, intensive attention

Indications for EEG examination:- epilepsy and other disorders of consciousness

- expansive intracranial processes

- sleep disorders

- encephalitis

- vascular brain injury

- intoxications

- craniocerebral injuries

- degenerative brain diseases

THE HEART

Cardyomyocites

• T-tubuls – diads

• Sarcoplasmic reticulum

• Nucleus, size, intercalated discs

• Contractile proteins (actin i myosine)

• Electrical junctionsGap junctions in intercalated discs – gap junctions

Intercalated discs

Action potential of cardiommyocites

• Plateau

– Fast sodium channels Na+

– Slow Ca++-Na+channels

– Slow potassium channels K+

prolongating the duration of

DEPOLARIZATION

delaying of

REPOLARIZATION

10x longer

contraction

The velocity of conduction

-Working muscle fibers 0.3-0.5 m/s

-Conductive muscle fibers 0.02-4 m/s

mV

- 100

- 80

- 60

- 40

- 20

0

20

250 ms

plateau

Na+

Ca++

K+

depolarization repolarization

Specialized musculature

Sinoatrial node – S-A node

Internodal pathways i interatrial pathway

Atrioventricular node – A-V node

Bundle of Hiss– left and right branch

Purkinje fibers

Special system for generating and conducting of impulses

• Generating rhythmical impulses

• Fast conduction of impulses through the cardiac muscle

S-A node

• Wall of the right atrium

• P-cells – pacemaker cells

• The head of cardiac rhythm- pacemaker

• f = 60-80 beats/min.

sinoatrial (S-A) node

Ca2+K+

Atrial pathways

• Anterior interatrial pathway

• Internodal pathways:

– Anterior Bachman’s

– Middle Wenckebach’s

– Posterior Thorel’s

internodal pathways

A-V node

• Posterior part of interatrial septum of the right

atrium

• Slowing the conduction of impulses –

synchronization of contraction

• f = 40-60 beats/min.

atrioventricular (A-V) node

Bundle of Hiss and Purkinje fibers

• Right and Left bundle branch – for right and left ventricle

• Up to around 1/3 of musculature of ventricles

• f = 15-40 beats/min.

left bundle

branchBundle of

His

Purkinje fibers

right bundle

branch

ECG

• The record of summary electrical activity of

cardiac muscle

The basics od ECG registration

- Tissues are electrical conductors

- Polarization of cell membrane

- The current flows from depolarized to non-polarized part, due to difference in potentials

• They registrate the oscillations in potential differences between positive and negative electrode

• 12 standard leads:

– 3 bipolar limb leads

– 3 unipolar limb leads

– 6 precordial leads

The Leads

The Leads

in frontal plane

BIPOLAR AND MONOPOLAR LEADS

Lead Positive electrode Negative electrode

I Left arm Right arm

II Left leg Right arm

III Left leg Left arm

aVr Right arm Left arm and left leg combined

aVl Left arm Right arm and left leg combined

aVf Left leg Right arm and left arm combined

The Leads

in horizontal plane

PRECORDIAL LEADS

Lead Pozitive electrode

V1 IV ICS right by sternum

V2 IV ICS left by sternum

V3 Middle between V2 and V4

V4 V ICS medioclavicular line virtuel heart center

V5 Middle between V4 and V6

V6 V ICS midaxillary line

Electrocardiography – ECG

P wave T wave

complex

Electrocardiography – ECG

Waves on ECG

• P wave: atrial depolarization

pozitivan je u standarnim odvodima i negativan u aVr

P

R

Q S

T

• QRS complex: ventricular depolarization (atrialrepolarization mimicked by QRS)

• T wave: ventricular repolarization

ventricular repolarization spreads in reversed direction than depolarization

• P-Q segment: complete atrial depolarization

- since the end of P wave, until the beginning of Q

Segments on ECG

P

Q

R

S

T

P

Q

R

S

T

• S-T segment: complete

ventricular depolarization

- since the end of S wave, until the beginning of T wave

P-Q interval

- Since the beginning of the P wave until the beginning of

the QRS: 0.12 - 0.21 s, depending on the heart rate

- The time between the beginning of atrial excitation until

the beginning of ventricular excitation komora

- The time of impulse conduction from SA to AV node

Intervals on ECG

Q-T interval

- Since the beginning of QRS complex until the end of T wave

- Time necessary for ventricular depolarization and repolarization

- Depends of heart rate:

of HR is 75 beats/min: 0,35-0,40 s

P

Q

R

S

T

P

Q

R

S

T

Extrasystole

• Premature beat- Premature Atrial Contractions (PACs) – additional P waves

– Premature Ventricular Contractions (PVCs) – bizzare QRS complexes

Characteristics:• Compensatory pause

• Post-extrasystole

- on ECG looks the same as the complex before extrasystole

- on mechanocardiogram has bigger amplitude

Extrasystole

S – systole

ES – extrasystole

PES – postextrasystole

CP – compensatory pause

CP

Regulation of heart pumping

autoregulation

Frank-Starling mechanism of the heart

neural regulation

sympathicus – heart pumping

parasympathicus – heart pumping

reflex regulation – the experiment of Goltz, for example...

humoral regulation

hormons: epinephrine, norepinephrine, T3, T4 – heart pumping

acetil cholin – heart pumping

K+ – stops the heart in relaxed state

Ca++ – stops the heart in contracted state

temperature

t C – heart pumping

t C – heart pumping

ECG Analysis

• Rhythm

• Axis

• Frequency

• Pathological signs: - cardiac arrhythmias

- Ischemia

- hypertrophy of left and right

heart

- electrolite dissorders

ECG Analysis

• During the standard speed of registration:

25 mm/sec (1mV= 1cm ):

• the smallest vertical square represents: 0.1 mV

• the smallest horizontal square represents 0.04 s, and bigger square 0.2 s

Rhythm

• Sinus – physiological

– P wave precedes to QRS complex

– Each P wave in the same lead is identical

Electrical axis of the heart

- Physiological axis: 59 (-20 -100 )

1. Recordings in standard leads (I, II, III) –

Einthoven’s law – R is the highest in II

2. Looking on leads I and aVf:

I lead aVF Axis

+ + normal axis

+ - left axis deviation

- + right axis deviation

- - extreme left axis deviation

Frequency

Physiological while resting: 60-80 beats/min

- depends on pacemaker

Ways to assess frequency

I way

1mm is 0,04 s

Count the number of PQRST in particular time interval – 5 or 6 s, then multplying it with 12 or 10, in order to get the valueof heart rate in one minute.

II way

300 150 100 75 60 50

ECG: sinus rhythm, normal axis, f 70/min,

without any pathological signs

The cardiac cycle

• Systole

– atriums 0.1s

– ventricles 0.3s

• Diastole

• atriums 0.7s

• ventricles 0.5s

Mutual systole of atriums and ventricles – doesn’t exist

Mutual diastole of atriums and ventricles – 0.4s

v atrium

ventricle

systole diastole

Pressures in heart chambers

Volume (ml)

Pre

ssure

(mm

Hg)

0

150

AB

E

D

C

Left ventricle

A – the beginning of diastole, mitral valves open – pressure is 0 mmHg

B – the beginning of systole, closing of mitral valves

C – the beginning of ejection phase, opening of aortic valves – pressure is 80mmHg

D – pressure rises up to 120mmHg

E – end of systole, closing of aortic valves

Pressures in heart chambers

Volume (ml)

Pre

ssure

(mm

Hg)

0

30

AB

E

D

C

Right ventricle

A – the beginning of diastole, tricuspid valves opened – pressure is 0 mmHg

B – the beginning of systole, closing of tricuspid valves

C – the beginning of ejection phase, opening of pulmonary valves – pressure is 7-

8mmHg

D – pressure rises up to 25mmHg

E – end of systole, closing of pulmonary valves

Hemodynamic parameters of heart

• Stroke volume – SV = 70-80 ml

• End-systolic volume – ESV = 40-50 ml

• End-diastolic volume – EDV = 110-120 ml

• Cardiac output – CO = f · SV 4-5 l/min

(COmax 20-30 l/min)

• Ejection fraction – EF = 60%

SV

EDV

Heart sounds

• 4 sounds

• Sound vibrations appear during the pumping of the heart

• Only closure of valves can be heard, not their opening

Phonocardiogram from I and II heart sound

I II

0.12s 0.08s

I heart sound

– Genesis is complex

• Valvular component – closure of AV

valves

• Mural component – contraction of

the walls of ventricles

– Systolic sound

– Lower tone

– Duration 0.12s

– Gradually appears and disappears

(crescendo-decrescendo typ)

– Best is heard on the projection of mitral

and tricuspid area on the chestwall

+

II heart sound

– Origin

• Closure of semilunar valves

– Diastolic sound

– Higher tone

– Duration 0.08s

– Suddenly appears and disappears

– Best is heard on the projection of

aortic and pulmonary area on the

chestwall

III and IV heart sound

• III heart sound originates due to vibrations of the

walls of ventricles during their fast load phase

(physiologicaly, common in children)

– early diastolic sound

• IV heart sound originates during the systole of

atriums

– late diastolic sound – usually a pathological sign

Auscultation of heart sounds

• Projections of areas on the chestwall

– Mitral area – IV or V ics left, 1cm medial to

medioclavicular line

– Aortic area – II ics right by sternum

– Pulmonary area – II ics left by sternum

– Tricuspid area – VI ics right by sternum

• Stenosis – contriction

of the confluence

murmur generates while the

blood flows through

constricted confluence

• Insufficiency – valves don’t close

the confluence entirely –

regurgitation

murmur generates when the

blood returns backwards after

the closure of the valve

Systolic and diastolic murmurs

Organic and functional genesis

I-VI degrees

Causes:

– Valvular disfunction

– Turbulence of blood flow

– Congenital anomalies

Heart murmurs

Pressures in systemic circulation

0

20

40

60

80

100

120

Left

ven

tricle

Great

veins

Pressures in pulmonary circulation

0

25

Rig

ht

Ven

tricle

pulmonary

viens

Measurement of arterial blood

pressure

• Directly and indirectly

Korotkoff methodRiva-Rocci method

TA 120/80 mmHg

TA hypertesion

TA hypotension

Capillary

• Cross-sectional area of capillaries

• Structure of capillaries

• Exchange of fluid in the capillaries

Fluid filtration across capillaries

• Difference in pressures

• Structure of capillaries

capillary

Oncotic pressure

40mmHg

18mmHg

28mmHg