Acta allergologica, ig^6, X,

From the Medical Clinic, State University, Utrecht, and from the Laboratory ofthe Willem Arntsz Hoeve in Den Dolder, The Netherlands

S T U D I E S ON H I S T A M I N E

INFLUENCE OF PHYSICAL EXERTIONON BLOOD HISTAMINE^

By

J. C. M. VERSCHURE, J. A. H. GOOSZEN, P. SJOUKES,

JOHANNA W. MAAS & TH. STRENGERS

I N T R O D U C T I O N

In the following experiments the blood histamine in normalsubjects was investigated before, during and after physicalexercise. Anrep & Barsoum (1935) discovered that duringcontractions of striated skeletal muscles histamine was set freein the venous blood—an observation re-affirmed by Anrep(1944) after Kwiatkowski (1941) had rejected the fact.Gabraoni (1950) also verified that after using the hand-muscles histamine was set free in the V. cubiti, detected onlyif the bloodsamples were taken immediately. Anrep, Barsoum,Talaat & Wieninger (1939) demonstrated that this histamineoriginates from a muscle depot. Anrep, Barsoum & Talaat(1936) demonstrated that extensive amounts of histamine alsooriginate from heart muscle. Code, Evans & Gregory (1938)doubted if histamine was formed by the heart, but Marcou(1939) confirmed the observation by' Anrep et al. Ambache& Barsoum (1939) demonstrated, that histamine is also setfree during contraction of smooth muscle.

' This investigation has been supported by a research grant from the "Neder-landse Organisatie voor Zuiver VS^etenschappelijk Onderzoek (Z.W.O.)".

137

METHODS AND RESULTS

The histamine determinations were carried out accordingto Code's method (1937) or according to the .micromodifica-tion of Gooszen, Donker & Strengers. (This series no. I I ) .

In a series of experiments we investigated whether a rise inblood histamine occurred after physical exertion. In a firstseries bloodsamples were withdrawn from the V. cubiti from10 normal males, after which they raced up and down 34stairs twice. Further samples were then withdrawn at once.The histamine values of both samples are given in Table I.

TABLE I

Normal males

I

2

3456

78

910

Histamine contentbefore physical exertion

in r/100 ml

6.44 . 0

3.89.0

6.1

8.8

5.75.0

7.72.8

Histamine contentafter physical exertion

in Y/lOO ml

8.0

11.8

9.89.88.1

9.812.3

344 . 0

2.1

Out of the 10 males, 7 were found to have an increasedlevel of blood histamine. In three cases the rise was quiteconsiderable, being greater than the experimental errors. Inthree cases a slight fall was observed.

A second series of experiments was carried out on 15 menand 7 women. They were asked to rest on a couch, during whichtime three blood samples were withdrawn. They were thenasked to raise and lower their legs a number of times keepingthem straight, during which time more samples were taken.Finally, during the following rest period, blood samples wereagain withdrawn. All blood samples were withdrawn from the

138

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10 Acta Allergologica, X, 3

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V. cubiti and all at intervals of 30 sec. The results of thehistamine determinations on the males are given in Table II.The results on the females in Table III. From Table III it isapparent tbat 3 of tbe females sbowed results quite differentfrom tbe otbers i.e. a bigb blood bistamine before tbe exerciseand a lower level during it. Statistical analysis tberefore wasonly carried out on tbe results obtained from tbe males.

Tbe bistamine levels found before, during and after tbeexercise were studied in order to see if tbey remained constantor if tbey sbowed any systematic increase or decrease. A trendtest (Kendall, 1948; Kaarsemaker en van Wijngaarden,1952; Sillitto, 1947), based on rankcorrelations, was tbere-fore applied to:

A. Observations before pbysical exertion.B. Tbe last observation before, togetber witb tbe observa-

tions during pbysical exertion.C. Tbe last observation before, togetber witb tbe observa-

tions during tbe second restperiod.

Tbe results are summarized in Table IV. (See for tbe in-dices used in tbe statistical analysis tbe scbema.)

From tbe table it appears that tbe blood bistamine risesduring pbysical exertion and falls after it. Tbere is also anindication tbat it rises just before tbe pbysical exertion started.Tbe levels before, during and after tbe exercise were next-

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141

TABLE IVTrend test on the hlood histamine values before {A), during (B), and after (C)

physical exertion.

Stot.Tail

probability

ABC

+ 7+— 24

8.1920.5211.28

4- 1-95+ 3-6i— 2.13

0.0s -t-

< 0.001 +0.03 —

TABLE VComparison of the hlood histamine values before (A), during (B) and after (C)

physical exertion, using Wilcoxon's Two Samples Test.

U —[U — | Tail

probability

B-AC-BC-A

— 25.520

I

11.96

"•738.gi

— 2.131.73o.ii

0.03 -f-0.08 —0.91

compared by means of Wilcoxon's Two Sampe Test. Tberesults of wbicb are given in Table V.

Tbis table sbows tbat tbe blood bistamine is bigber duringpbysical exertion tban before it, tbat a sligbt difference existsbetween tbe levels during and after pbysical exertion and tbattbere is no difference between tbe levels before and after tbepbysical exertion.

A furtber analysis was tben carried out and tbe following"differences" were computed:

a. Tbe difference between tbe second and first observa-tions during tbe period before pbysical exertion. Tbisdifference being a measure of tbe rise at tbe start of tbeexperiment.

b. Tbe difference between tbe last observation during tbeperiod before pbysical exertion and tbe last observationbut one during tbat period. Tbis difference being ameasure of tbe rise directly before pbysical exertion.

142

c. The difference between the first observation after thestart of the period of physical exertion and the last onebefore it. This difference being a measure of the riseat the start of the period of physical exertion.

The hypothesis that the differences c—a and c—b respectivelyhave the same probability distribution, was investigated bymeans of the Test of Symmetry (Hemelrijk, 1950). Theresults are given in Table VI.

TABLE VITest of symmetry to compare rises before and at start of physical exertion.

c-ac-b

n

15

r

g7

Tail probability

I 6 0.02 —

I 4 > o.io —

From these results we may conclude that the rise at the be-ginning of the experiment is greater than the rise at the startof the period of physical exertion and that there is no reasonto accept a difference between the values just before and justafter the start of the period of physical exertion.

To investigate whether a demonstrable increase occurs bothat the beginning of the experiment and at the start of theperiod of physical exertion, the test of symmetry (Tj) wasapplied to the differences a and c separately. (See Table VII.)

No increase in blood histamine was indicated in either case.We must therefore conclude that the increases before and

TABLE VIITest of symmetry Tt to investigate increase of blood histamine at the beginning

of the experiment and at the start of the period of physical exertion.

Tail probability

a 13 7 2 I >o . io +

c 13 6 2 3 > o.io —

14.3

during the period of physical exertion, shown in Table IVwere secured gradually.

Starting physical exertion did not appear to have any effecton the blood histamine. This perhaps may be explained bythe hypothesis that histamine is set free by the contractingmuscle. This histamine will only appear after a lapse of time,depending on the rate of circulation.

The influence of cessation of physical exercise on the bloodhistamine was next investigated. Table IV shows a fall in theblood histamine after physical exertion. Table V shows nodifference between the levels before and after physical exer-tion. In the subsequent analysis the following "differences"were computed:

d. The difference between the last observation during theperiod of physical exertion and the last observation butone during that period. This difference being a measureof the rise at the end of the period of physical exertion.

e. The difference between the last observation during theperiod of physical exertion and the first observation inthe second restperiod. This difference being a measureof the decrease after the cessation of physical exertion.

These values were tested by applying the test of sym-metry T2 to the differences d-e, while the same test was alsoapplied to the differences e themselves. The results will befound in Table VIII.

TABLE VIIITest of symmetry Ti to compare ihe rise at the end of the period of physicalexertion with the decrease immediately after cessation of physical exertion.

d-ee

n r

7g

u

I

2

Tail probability

0.04 —

0.05 —

It appears that the rise just before the end of the physicalexertion period is less than the fall just after it. There is also

144

an indication that a decrease does occur on cessation of physicalexertion. The question arises whether from this decrease weare justified in assuming that physical exertion does indeedcause an increase of blood histamine. It is possible that theincreased level found at the start of the experiment and whichcannot be attributed to physical exertion, gradually fell tobelow normal, in an effort to recover the equilibrium, and thatthe moment of decrease coincided with the end of the periodof physical exertion. We therefore investigated whether theblood histamine was still raised during the latter part of theperiod of physical exertion, by applying the test of symmetryT2 to d. A tailprobability > o . i o was found. Towards the endof the period of exertion therefore no increase could be de-monstrated.

The rise in histamine during the physical exertion periodwas analysed further by comparing the histamine values ofthose people, on whom observations were made over a longperiod (long series), with the histamine values of those, onwhom observations were made over a short period (shortseries). The total number of observations was counted hereas the number of observations from the start of the experi-ment upto and including the end of the period af exertion. Inthe one group only 7 observations or less had been made oneach person, in the other group, more than 7. In each weinvestigated whether the difference d (the difference be-tween the last observation and the last observation but oneduring the physical exertion period) was positive or nega-

TABLE IXDistinction between short and long series of observations from the beginning of

the experiment upto and including the end of physical exertion period.

d

longshort

Total

+

r6

7

-

53

g

Total

6

9

15

145

tive. The results of these computations are summarized inTable IX.

By means of a 2 X 2 table we checked the hypothesis thatdiscernment between long and short observational series isnot dependant on discernment between increase or decreaseat the end of the exertion period. The theory was also checkedagainst an alternative hypothesis that the probability of adecrease at the end of the exertion period is greater in longseries than in short series. The one sided tailprobability turnedout to be = 0.08. We thus found a slight indication thatcontinuous physical exertion over a long period, results in adecrease of histamine before the exertion ceases.

In order to investigate the behaviour of the histamine inde-pendantly from physical exertion, a distinction was again madebetween the short and long series and also between the positiveand negative differences between the 7th and 8th observa-tions. (In the long series, this differences fell during theexertion period, in the short series it fell between the lastobservation during, and the first observation after the physicalexertion period. In cases with less than 7 observations, thislatter difference was also used.) Results will be found inTable X., . . . •

TABLE XDistinction betiueen short and long observational series and positive and negative

differences betiueen the 7th and 8th observations.

Total

longshort

Again we find a one sided tailprobability of 0.035. I" thiscase it indicates that when physical exertion was continuedbeyond the 7th observation, the blood histamine was moreoften increased than in those cases who stopped after or be-fore 7th observation. Analysing further, but counting thelength of the series from the beginning of the exertion period

146

instead of the beginning of the experiment and then againdistinguishing hetween long and short series, the applicationof the 2X2 table resulted in one sided tailprohabilities of0.02 and 0.76. The first result supports our previous con-clusions that in long series the decrease occurs already heforethe end of the exertion period. The large tailprobahility in thesecond result, might he due to the fact that hy shifting thehoundary hetween the long and short series, it has ap-proximated the end of the physical exertion period so closelythat in most cases a decrease had already occurred.

Finally in order to investigate the trend of the decreaseafter the period of physical exertion, the trend test was ap-plied to the ahove ohservations hut this time omitting the lastobservation during the exertion period. The results will hefound in Tahle XI.

TABLE XITrend test to investigate the decrease of histamine after physical exertion

omitting the last observation during the exertion period.

hot

— 3 7.60

^tot.0

0-39

Tailprobability

0.70 —

The tailprobability was found to increase from 0.03 to 0.70when omitting the last observation during the period of exer-tion (cf. Table IV). This indicates that the fall in histamineafter physical exertion occurs for the greater part directlyafter the exertion has ceased.

As the increase in the tailprohahility is partly caused hy thedecrease of the numbers of ohservations, the test of symmetrywas also applied to the differences hetween the first and lastobservation after physical exertion. A tailprohahility of> o. 10 (—) was found. No decrease was found therefore,to support our previous conclusion.

147

D I S C U S S I O N

On reviewing the results of the statistical analysis, we findthat the conclusions formed by different tests are all com-patible. Whilst some tests, however, give a dear answer to acertain question, others give only an indication.

It appears, that the blood histamine rises before and duringphysical exertion. After the exertion has ceased, the histaminedecreases, but in some cases this decrease occurs even earlier,probably in those cases with a long period of physical exertion.The decrease after the exertion period is probably a quick one.These last conclusions are in accordance with the opinion thatthe normal organism compensates increases of histamine ra-pidly (see no. I of this series).

The fall in histamine during a long period of exertion couldbe explained perhaps by presuming that histamine is set freefrom muscle, bound histamine changing into free, but thatduring longer periods of muscular exertion some depletionof the bound histamine occurs. Here we refer to the investiga-tions of Strengers and Maas (this series no. IV) who foundthat in muscle of the guinea-pig the bound histamine isprobably completely changed into free histamine during con-tractions in the agonal phase. Referring to the increasedhistamine during physical exertion we were unable to ascertainwhether this gradual increase results from physical exertion,or is a continuation of the increase already observed beforethe exertion period. But referring to the investigations men-tioned in the introduction (this series no. I) our opinion is,that physical exertion does have a direct influence on thehistamine level. Most striking is the established increase duringthe period before physical exertion. We are bound to concludethat a psychic factor is the cause here of the "auslosend Mo-ment" of histamine mobilisation. Relating to this we wouldlike to draw attention to the irregular behaviour of the his-tamine level before physical exertion in three of the sevenwomen under investigation. We refer also to Gooszen (thisseries no. VI) who ascertained that in a number of schizo-phrenic patients rapid changes in the histamine level occurred.

148

SUMMARY

The effect of physical exertion on blood bistamine wasinvestigated.

The blood histamine rises just before (due to a psycbicfactor) and during pbysical exertion. The blood histaminedecreases after the exertion has ceased, and in some cases tbedecrease occurs even earlier. The decrease after the exertionperiod is probably a quick one.

REFERENCES

Ambache, N. & Barsoum, G. S.: J. Physiol., p(5, 139, 1939.Angrep, G. F. & Barsoum, G. S.: ]. Physiol., 55, 409, 1935.

- - & Tataat, M.: J. Physiol., 5d,+3i, 1936.- - - & Wieninger, E.: J. Physiol., p5, 148, 1939 and 9^,240,1939.- - Salama, S. & L. Spuidan: ]. Physiol., loj, 297, 1944.

Code, C. F.: J. Physiol., ^p, 257, 1937.- Evans, C. L. & Gregory, R. A.: J. Physiol., 5<5, 431, 1938.

Gabraoni, B. & Roy, J.: Egypt. Med. Assoc, 33, 677, 1950.Hemelrijk, J.: Proc. Kon. Ned. Akac. v. Wet., 5^,945, 1186, 1950.Kaarsemaker, L. Sc IVijngaarden, A. van: Rapport R 73 of the Computation De-

partment of the Mathematical Centre, Amsterdam, 1952.Kendall, M. G.: Bank Correlation Methods, London, 1948.Kiviatkowski, H.: J. Physiol.,/00, 147, 1941.Marcou, I.: C. R. Soc. Biol., 130, 573, 1939.Sillitto, G. P.: Biometrica, 34, 36, 1947.