9
Cancer and Human Liver Catalase* EDWARDE.MASON,TING-FONGCHIN,YAOW.Li,ANDSIDNEYE.ZIFFREN (Department, nf Surgery, Stale University of Iowa, College of Medicine, lou-a City, loica) SUMMARY Data presented in the present paper indicate that human liver catalase depression is related to weight loss. A statistical study was first made to determine the catalase activity in correlation of the iodotitrimetric and spectrophotometric methods for biopsy and autopsy samples from cancer and cancer-free patients. Cancer patients had a 22 per cent lower liver catalase activity than cancer-free patients. A two-by-two factorial analysis of variance comparing presence or absence of cancer and weight loss revealed that weight loss accounted for the catalase depression and that, when correc tion was made for the weight loss effect, no additional cancer effect was seen. Distribu tion of catalase in subcellular fractions was also studied and failed to show any sig nificant cancer effect but did demonstrate the relationship between weight loss and catalase depression in the soluble fraction. No effect of sex was observed, which is con sistent with Adams' observations in laboratory animals that, when the male of a species does not show an increased liver catalase as compared with female, no sig nificant depression of liver catalase is observed (2). Studies of liver catalase depression by cancers of men were pioneered in 1910 by Blumen thai and Brahn (6). They reported that the liver catalase activity was very low in human beings who had died as a result of various forms of cancer. Experi mental work prior to 1941 has been summarized by Greenstein (11) and has amply demonstrated that the peroxide-splitting enzyme, catalase, is con siderably reduced in the liver of some strains of mice and rats bearing tumors. The decrease in liver catalase in susceptible animals is progressive with the growth of the tumor. There have been re ports that in some strains of animals depression of catalase did not occur despite rapid tumor growth to a large size (4). When an effect is observed the tumor must grow rapidly and reach a size equal to 5 per cent of the animal's total body weight before the effect can be detected (10). Recently Nakahara and Fukuoka (15) isolated a heat-stable, protein-like catalase-depressing material from human tumors and demonstrated its effect in mice. Nakagawa (14) has isolated a liver catalase-depressing substance from the urine * This work was supported by the J. B. Phillips Memorial Grant for Cancer Research from the American Cancer Society. Appreciation is expressed to Prof. E. F. Mason for proofreading and checking of statistical calculations. Received for publication May 4, 1960. of a cancer patient and the gastric juice of patients with stomach cancer. A comparison between the liver catalase activity of patients with malignant tumors and patients with gastric or duodenal ulcer was reported in 1958 by Kiyota (13). They found a lower catalase activity in the cancer patients but did not evaluate the possible nonspecific ef fect of poor nutrition. A nutritional effect in liver catalase activity of animals has been reported by Van Pilsum (17) in 1957. The specificity of de pression of liver catalase due to cancer in man is therefore still open to question, since malnutrition alone could cause the mild depression observed. It has also been observed by Adams (1) that only the liver catalase of animals which is increased by testosterone is depressed by cancer. The present work was undertaken in three rather distinct phases. Initially an investigation was made to find out whether there was any differ ence between patients with cancer and patients who were operated upon for noncancerous condi tions. At the same time the methods used were evaluated for reliability. Secondly, another im portant factor was examined, namely, the presence of a greater frequency and per cent of weight loss in cancer patients as compared with noncancer patients. This was an amplification in which a two- by-two factorial analysis was used, and the two 1474 Research. on February 2, 2019. © 1960 American Association for Cancer cancerres.aacrjournals.org Downloaded from

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Page 1: Cancer and Human Liver Catalase*cancerres.aacrjournals.org/content/canres/20/10_Part_1/1474.full.pdf · Cancer and Human Liver Catalase* EDWARDE. MASON,TING-FONGCHIN, YAOW. Li, ANDSIDNEYE

Cancer and Human Liver Catalase*

EDWARDE. MASON,TING-FONGCHIN, YAOW. Li, ANDSIDNEYE. ZIFFREN

(Department, nf Surgery, Stale University of Iowa, College of Medicine, lou-a City, loica)

SUMMARY

Data presented in the present paper indicate that human liver catalase depression isrelated to weight loss. A statistical study was first made to determine the catalaseactivity in correlation of the iodotitrimetric and spectrophotometric methods forbiopsy and autopsy samples from cancer and cancer-free patients. Cancer patients hada 22 per cent lower liver catalase activity than cancer-free patients. A two-by-twofactorial analysis of variance comparing presence or absence of cancer and weight lossrevealed that weight loss accounted for the catalase depression and that, when correction was made for the weight loss effect, no additional cancer effect was seen. Distribution of catalase in subcellular fractions was also studied and failed to show any significant cancer effect but did demonstrate the relationship between weight loss andcatalase depression in the soluble fraction. No effect of sex was observed, which is consistent with Adams' observations in laboratory animals that, when the male of a

species does not show an increased liver catalase as compared with female, no significant depression of liver catalase is observed (2).

Studies of liver catalase depression by cancersof men were pioneered in 1910 by Blumen thai andBrahn (6). They reported that the liver catalaseactivity was very low in human beings who haddied as a result of various forms of cancer. Experimental work prior to 1941 has been summarized byGreenstein (11) and has amply demonstrated thatthe peroxide-splitting enzyme, catalase, is considerably reduced in the liver of some strains ofmice and rats bearing tumors. The decrease inliver catalase in susceptible animals is progressivewith the growth of the tumor. There have been reports that in some strains of animals depression ofcatalase did not occur despite rapid tumor growthto a large size (4). When an effect is observed thetumor must grow rapidly and reach a size equalto 5 per cent of the animal's total body weight

before the effect can be detected (10).Recently Nakahara and Fukuoka (15) isolated

a heat-stable, protein-like catalase-depressingmaterial from human tumors and demonstratedits effect in mice. Nakagawa (14) has isolated aliver catalase-depressing substance from the urine

* This work was supported by the J. B. Phillips MemorialGrant for Cancer Research from the American Cancer Society.Appreciation is expressed to Prof. E. F. Mason for proofreadingand checking of statistical calculations.

Received for publication May 4, 1960.

of a cancer patient and the gastric juice of patientswith stomach cancer. A comparison between theliver catalase activity of patients with malignanttumors and patients with gastric or duodenal ulcerwas reported in 1958 by Kiyota (13). They founda lower catalase activity in the cancer patientsbut did not evaluate the possible nonspecific effect of poor nutrition. A nutritional effect in livercatalase activity of animals has been reported byVan Pilsum (17) in 1957. The specificity of depression of liver catalase due to cancer in man istherefore still open to question, since malnutritionalone could cause the mild depression observed. Ithas also been observed by Adams (1) that only theliver catalase of animals which is increased bytestosterone is depressed by cancer.

The present work was undertaken in threerather distinct phases. Initially an investigationwas made to find out whether there was any difference between patients with cancer and patientswho were operated upon for noncancerous conditions. At the same time the methods used wereevaluated for reliability. Secondly, another important factor was examined, namely, the presenceof a greater frequency and per cent of weight lossin cancer patients as compared with noncancerpatients. This was an amplification in which a two-by-two factorial analysis was used, and the two

1474

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MASON et al.—Cancer and Human Liver Catatase 1475

variables, cancer-no cancer and weight loss-noweight loss, were subjected to an analysis ofvariance after an accumulation of sufficientreplications of appropriately selected patients toprovide equal numbers of patients in each of thefour groups. Finally, the distribution of humanliver catalase in the subcellular fractionations wasinvestigated in relation to the effect of cancer andweight loss. Also, an analysis was made of therelationships between the human liver catalaseactivity and sex. All these findings are consistentwith experimental work in certain strains ofanimals in which liver catalase is not directly influenced by cancer. The analysis of data leading tothis conclusion and the interpretation of thesefindings comprise the following presentation.

EXPERIMENTAL AND RESULTSPART A

A study of arithmetic means and variations ofliver catalase activity in cancer and noncancerpatients as compared with the statistical correlation of two analytical methodsSelection of patients and samples.—Liver bi

opsies were obtained from patients whose operative procedure involved exposure of the upperabdomen. Livers from autopsies were obtainedwhen patients were examined within 6 hours ofdeath. Samples were obtained from patients whodid not have primary liver disease and from areascontaining no metastatic tumor. The specimenswere chilled immediately and taken to the laboratory, where a 5 per cent homogenate was preparedin 0.05 M phosphate buffer at pH 6.8 with aPotter-Elvehjem homogenizer surrounded bycrushed ice.

Methods used in analysis.—

1. The titrimetric method used for thesestudies is a modification of the method of Sumnerand Dounce (7). A 0.05-ml. aliquot of the 5 percent homogenate was added to 50 ml. of 0.1 Mhydrogen peroxide buffered with a 0.0067 Mphosphate buffer at pH 6.8 and in an ice-cooledbeaker. Stirring was done with a Teflon-coveredmagnet in a moving magnetic field. Samples of5 ml. volume were withdrawn from the reactionchamber at 0, 1,2, and 3 minutes and injected into5 ml. of 2 N sulfuric acid. To each of the samples10 ml. of 10 per cent potassium iodide and 1 dropof 1 per cent molybdic acid were added. Exactly3 minutes after the addition of the iodide, titrationwas carried out with 0.005 M thiosulfate, with afew drops of starch added toward the end of thetitration to improve the end-point. A constantartificial lighting was used, since sunlight wasobserved to affect the titration.

2. For the spectrophotometric method (5) aBeckman Model DU spectrophotometer was usedwith 1-cm. light path quartz cuvettes. The rate ofdisappearance of hydrogen peroxide was measuredat 240 imi by change in optical density. As a preliminary step, two cuvettes were prepared with 3ml. of an appropriately dilute homogenate in 0.5 Mphosphate buffer of pH 6.8. At zero time 1 ml. ofa 3.5 X 10~2M solution of hydrogen peroxide in

buffer was added to one of the cuvettes and 1 ml.of buffer was added to the other as a blank.Optical density measurements were then taken at10-second intervals for 4 minutes. The unit ofcatalase activity used is defined by this equation:

TT ¿»3 * Id r /-l-l

U is the unit of catalase activity per minute pergram of fresh liver sample.I0 is the initial concentration of the thiosulfate.It is the concentration of the thiosulfate at tminutes.T is reaction time in minutes.C is the concentration of homogenate.

From Table 1 the mean catalase activity for theseventeen biopsy samples was found to be 22.5 percent lower in cancer patients than in the patientshaving no cancer. The correlation between thesetwo methods in 24 samples (autopsy included) was0.90. It was 0.79 in the seventeen biopsy samples.This indicates a high degree of correlation betweenthese two methods. Although the correlation ishigher when the autopsy data are included, thecorrelation coefficient of 0.79 was sufficiently highin the biopsy data to warrant use of either methodfor studies of catalase activity in human liversamples obtained from living patients with andwithout cancer. Subsequent analyses were therefore carried out with the titrimetric method alone.

PART BComparison of cancer and weight loss as they

relate to liver catalase activity with use ofa two-by-two factorial analysis

In Part A we found that the liver catalaseactivity was 22.5 per cent lower in the cancerpatients than in the patients having no cancer.The question remained as to the cause of this depression in cancer patients. Body weight loss wasoften present as an evidence of poor nutrition. Todetermine the relationship of liver catalase activity to body weight loss and to cancer, the patientswere divided into four groups. These groups ofpatients were selected as having: (a) less than 10per cent body weight loss and no cancer, (b) more

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1476 Cancer Research Vol. 20, November 1960

than 10 per cent body weight loss but withoutcancer, (c) less than 10 per cent body weight losswith cancer, and (¿)more than 10 per cent bodyweight loss with cancer. Determination of bodyweight loss was by the history recorded in thepatient's hospital record. This usually consisted ofboth the "present" and "usual" weight and the

absolute weight loss plus the time interval ofweight loss. Because of variations in body build,obesity, and duration of weight loss, it was considered desirable to simply divide the patientsinto groups as they fell above or below an arbitrary 10 per cent body weight loss and thus toconsider weight loss as a categorical rather thanan ordered variable. The procedures for thebiopsy collection and homogenate preparationwere the same as in Part A, but only the iodotitri-metric method was used for determination ofcatalase activity. Some data from the earlier studywere used, as indicated by the identical patients'

numbers in the tables. The additional patients

were selected preoperatively on the basis of presence or absence of cancer or body weight loss, aswas required to supply an equal number of reiterations in the deficient groups for the projected two-by-two factorial analysis.

The data used for the analysis of variance areshown in Table 2, and the actual analysis is shownin Table 3. It is assumed that these are randomsamples from a normal population and that thevariance is the same for each portion of this normalpopulation. There are two columns in whichcancer and noncancer patients are compared andtwo rows wherein weight loss and no weight lossare compared. The final analysis culminates in anF value of 4.25 with which these statistics must becompared to determine their significance. F.95 (1,20) = 4.25 (8) indicates that if any of the Fratios are greater than 4.25 they are significantat a confidence level of 95 per cent. It is, therefore, apparent that the two-row means with an Fratio of 10.67 were significantly different, or in

TABLE1DATAFORCOMPARISONOFLIVERCATALASEACTIVITYINCANCERANDCANCER-FREEPATIENTS

Caseno.12345(i789101112131415161718192021222324Age6154466250807768717049687954597378837836812«8282SexMMFMFMFMMFMMFMFMMMFFMMMMPATIENTS'TumorwtBody

wt.0.260.530.7500.0000.10¿02050à

302.503.00WENTIFICATIONWt.l«sBody

wt.001.55Ó3163819•j1102231018?tttttDiagnostaPeripheral

vasculardiseaseGastriculcerDuodenalulcerDuodenalulcerCholecystitisDuodenal

ulcerMeanof biopsies,noncancerGastric

carcinomaEsophagealcarcinomaEsophagealcarcinomaRectaladenocareinomaColoniecarcinomaAbdominalcarcinomatosisGastriccarcinomaPancreaticcarcinomaPancreaticcarcinomaGastriccarcinomaGastric

carcinomaMeanof biopsies,cancerArteriosclerosisAcute

tracheobronchitisMultipleabscessesHip

fracture,diabetesBacterialendocarditisMean

of autopsies,noncancerGastric

cancerUrinarybladdercancerMeanof autopsies, cancerCATALASETitrimetric221323231280231205*249

±44205131145153213162133ins261214170*179±41140ias81191116«142

+4710142*72

+ 42ACTIVITYSpectro-photometric228264228247211156•222

±37202122192187230194158151208182180*183±3012514496173115•131+2910855*82

+ 37

* Arithmetic mean of groups ±standard deviation (root mean square deviation),

t Autopsy samples.

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MASON et al.—Cancer and Human Liver Catalase 1477

other words there was a relationship betweenweight loss and liver catalase activity. The columnmeans representing a comparison between cancerand noncancer were not significantly different.Since only weight loss had an effect, no interaction between weight loss and cancer was to beexpected, and none was found.

PARTCSubcellular distribution of catalase in patients

with the variables cancer-no cancer andand weight loss

Although we can be 95 per cent sure that arelationship between weight loss and liver catalasedoes exist, a statement that cancer does notspecifically cause a decrease in liver catalase cannot be made with the same degree of confidence.This is inherent in the nature of the statement andin the statistical methods. All one can say is thatno relationship could be found. Because it is generally believed that toxohormone does depress human liver catalase, it seems important to look forsuch an effect by making use of a more sensitiveexperimental technic. Since the soluble (non-particulate, nonsedimentable) catalase is reportedto be the only portion of liver cell catalase depressed early and by the smaller tumor growthsin laboratory animals (2, 3, 18), an investigationwas made of the activity distribution of catalasein subcellular fractions of human liver, again including patients selected with the variable cancerand weight loss for study.

In this experiment a PVP'-sucrose buffermedium (19) (10 per cent polyvinylpyrrolidonand 20 per cent sucrose, at pH 6.8) was used forhomogenization, and 0.1 per cent triton-XlOOwas added after fractionation to assure dissolutionof all the catalase-containing particles. The 5 percent homogenate was centrifuged at a speed ofIS.OOOX? for 30 minutes. The two fractionswere then separated, and each was made up tothe original volume. The titrimetric method wasused for determination of catalase activity in thewhole homogenate and in each of the fractions.The data indicating the distribution of humanliver catalase activity are shown in the Table 4.

Table 4 shows no cancer-specific depression ofeither the soluble supernatant catalase or the sedi-mentable particulate catalase. Recovery was only90.7 per cent in the patients who had both cancerand weight loss. The instability of the enzymeseemed to be slightly greater in the sediment thanin the supernatant. An analysis of variance was

1PVP donated by Farbenfabriken Bayer AG, Germany:molecular weight about 70,000.

carried out with the data from column C (Table 4)and again showed a significant (90 per cent confidence) relationship between weight loss andcatalase activity but no evidence of a selective orspecific tumor-produced agent which actively depressed the soluble portion of human liver catalase.The combination of cancer and weight loss wasassociated with a higher mean soluble catalaselevel—96—thanwas seen in patients with weightloss and no cancer—-88.These figures are not significantly different and cannot be used to provethat cancer increases liver catalase in patientswith weight loss, but they do support the earlierfinding that cancer has no significant depressiveeffect on the catalase of human liver.

A Mocancer biopsyA Cancer biopsyO No cancer autopsy•Cancer autopsy

40 80 120 160 200 240 280 320Units by Tilrimetric Method

CHART1.—Aplot of biopsy and autopsy data for cancerand cancer-free patients with the titrimetric method on theabscissa and the spectrophotometric method on the ordinate.Since the correlation coefficient, r = \/b b', where 6 and 6'are estimates of the regression lines of K on A' and X on Y,

these regression lines were drawn with the biopsy data usedalone (r = 0.79) and for all the data (r = 0.90).

In those strains of laboratory animals whichshow no depression of liver catalase by cancerthere is also no difference in catalase activity between the two sexes (2). Cancer-free patients fromthe present study were, therefore, paired according to weight loss into male and female groups. Itwas hypothesized that the means of the twopopulations are equal, and this was tested with thestatistic "t" (8) :

Zi(male) - X2 (female)+ U/AT2)

,-2) = -2.179;

-2) = +2.179

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1478 Cancer Research Vol. 20, November 1960

Since the observed t fell between the limits expected when an a of 0.05 was used, there is noreason to doubt that men and women have thesame level of liver catalase activity.

DISCUSSIONThe above experimental data suggest that

cancer depresses human liver catalase indirectlythrough the malnutrition produced in the host.That the liver catalase activity in cancer patientsis usually lower than in cancer-free patients is confirmed by Part A of the present study, but thecause of the lowering cannot be assigned to a

specific effect of cancer. Usually cancer is accompanied by weight loss, as is illustrated in PartB of this study. The patients having more than 10per cent body weight loss but without cancershowed a significant lowering of liver catalase.By contrast, the patients with cancer but withoutweight loss showed no significant liver catalasedepression. As a result of these analyses whichtake into consideration the additional variable,weight loss, it is necessary to reject the hypothesisof Blumenthal and Brahn and of Kiyota, neitherof whom considered body weight loss.

From these results, a new hypothesis is sug-

TABLE 2

DATAOFLIVERCATALASEACTIVITYINPATIENTSCLASSIFIEDACCORDINGTOPRESENCEORABSENCEOFCANCERANDWEIGHTLoss

PATIENTS*IDENTIFICATIONCase

no.AgeSexWt.

lossBody

wt.DiagnosisCATALASE

ACTIVITY

Group 1—< 10% body weight loss and no cancer

5i1Ì52627505461586649FMMMMM000000060709Group

2—> 10%body28293303132485546844949MMFMMF111215162234CholecystitisGastric

ulcerPeripheralvasculardiseaseGastric

ulcerDuodenalulcerPeptic

ulcerMeanweight

loss but withoutcancerGastric

ulcerDuodenalulcerDuodenalulcerPyloricobstructionObstructedstomachAbdominal

anginaMean231323221237228256*249

+3822023923116117896*188

+ 52

Group 3—< 10% body weight loss with cancer

331114343515744954737759MMMMFF(100103050609Group

4- >10%3616913378487371795968FMMFFM171819223038Gastric

carcinomaColoniecarcinomaPancreaticcarcinomaGastriccarcinomaColoniecarcinomaPancreatic

carcinomaMeanbody

weight loss withcancerColonie

carcinomaGastriccarcinomaEsophagealcarcinomaGastriccarcinomaGastriccarcinomaEsophageal

cancerMean166213183232•220261*213±34166214145133204131»166+ 35

' See footnote *, Table 1.

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MASON*et al.—Cancer and Human Lirer Catatase 1479

gested—that cancer has a very mild (if any) specificeffect on human liver catalase activity. This doesnot mean that human cancers do not produce toxo-hormone, since Nakahara and Fukuoka have isolated a catalase-depressing material, toxohormone,from human tumors and demonstrated its effect inmice. From Adams' series of papers it appears

that in susceptible strains of animals the action ofthe specific cancer-produced, liver catalase-in-hibiting material is to interfere or compete withthe testicular and adrenal hormones in theirstimulation of increased catalase production. If astrain of animals has the same liver catalaseactivity regardless of the level of testicular andadrenal hormones, then no cancer catalase-depressing effect is to be expected. The human subject, showing no sexual difference in liver catalaseactivity, is like those strains of laboratory animalsin which liver catalase is not susceptible to depression by toxohormone.

Additional support for this new hypothesis wasfound in the study of the distribution of humanliver catalase in subcellular fractions. Price (16)points out that, if cancer affects liver catalase, thedistribution will vary in fractions from cancer ascompared with cancer-free animals. He observedthat the insoluble fraction was not reduced in theliver of tumor-bearing animals, whereas thecatalase of the soluble fraction was reduced to

less than one-third that found in the corresponding fraction from normal liver. In our experiment,we found no significant effect of cancer on thedistribution of human liver catalase.

In a later paper Price (9) showed that the appearance of activity in the soluble fraction of micewas an artifact, since in the PVP-sucrose medium80-99 per cent of the activity was in the particu-late fraction, depending on the pH of the homog-enate. When catalase was obtained directly fromthe isolated part ¡culatea uniformly high specificactivity is found. There is less enzyme in the liverof tumor-bearing animals, but qualitatively it isidentical to that found in normal animals. Adamsrecently has again raised the question of distribution of catalase and gives data to indicate thatthere is a decrease in permeability of the membrane (3) of catalase-containing particles as a result of cancer. It was because of these reports thatfractional studies were undertaken and that PVP-sucrose medium was used. Since approximately50 per cent of human liver catalase was found assoluble, nonparticulate catalase, additional comparative studies were carried out in other specieswith the same homogenate medium and conditions(such as pH, centrifugal force, time and temperature for homogenization, etc.) The results weremost interesting and will be reported in detail in aseparate paper; there was a striking difference in

TABLE 3

STATISTICALANALYTICALDATAOFHUMANLIVERCATALASEACTIVITYINTWO-BY-TWOFACTORIALANALYSIS:CANCER-NOCANCERANDBODY

WEIGHTLoss-No BODYWEIGHTLoss

A: DATA OF TOTAL»

< 10% body> 10% bodyweight

lossweight lossNo.

cancer1,496

01,126.02,622.0Cancer1,276.0994.02,270.0Tot»!2,772.0¿.11904,891.0

B: ANALYSISOF VARIANCE;

How meansColumn meansInteractionSubtotalWithin

groups

TotalSum

ofsquares17,670

5,40049023,50033

,05056.550D.f.*1

1132023Mean

squnre17,670

5,4004301,652K

rati«10.67

3.270.20F

95 il, 20)= 4.35

* Degrees of freedom.

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1480 Cancer Research Vol. 20, November 1960

the distribution of catalase in the liver cells ofdifferent species. In our own laboratory 97 percent was sedimentable in mice at 18,OOOX<7,compared with 50 per cent in man. There was alsoan inactive sedimentable fraction which variedgreatly among species and was released or activated by triton-XlOO.

The relationship of body weight loss and livercatalase depression observed in this study is probably due to malnutrition. Nutritional effects havebeen documented in the laboratory animal. Iron,magnesium, copper (12), isoleucine, tryptophan,and phenylalanine (17) are all essential to catalase

formation, and other substances are probably alsoessential.

In conclusion, human liver catalase did showa demonstrable depression by cancer, but it can beexplained on a nonspecific basis. The distributionof catalase in human liver was different than intoxohormone-responsive animals. There was nosexual difference in liver catalase activity in man.These latter two findings help to correlate the findings in man and those in experimental animals.Although human liver is relatively insensitive totoxohormone, this does not invalidate the findingof others that toxohormone is produced by human

TABLE 4SUBCELLULAR DISTRIBUTION' OF LlVER CATALASE IN CANCER-FREE AND CANCER PATIENTS

COMPAREDWITHBODYWEIGHTLoss

PATIENTS'IDENTIFICATIONCaseAgeSexWt.

lo»«Bodywt.DiagnosisCATALASE

ACTIVITYOrigi

nalhomog-enate

ASedi

ment18000X»BSupernatant18000XICSTATISTICSDistribution5

xiooA- XIOOARecoveryB+C

—— XIOOA

Group 1—<10% body weight loss

38394041425835714964FFFMF00099GallbladderstoneGallstonesGallstonesPepticulcerPyloric

obstruction256265248244261115130142138132139131129107114«124+ 134549575150*505449524444*49999910»9594*99

Group 2—>10% body weight less

43444546474857497749MFMMF11U151734GastriculcerGallbladderstonesObstructed

stomachPseudotumorSuperior

mesentericarteryocclusion1992951632148987162761193497121768959»88

±234358465539*484941474267*4992999397105*97

Group 3—< 10% body weight loss

48495051525874647759MMFFM00355PulmonarycancerGastriccarcinomaPulmonarycarcinomaColoniecarcinomaColonie

carcinoma3751472422032121367314711110021768105105109»121+ 183650615547»505846445153*509496104106101•100

53545556577063794876FMFFM1216161727Group

4—>10%Gastric

carcinomaGallbladdercarcinomaPulmonary

carcinomaColoniecarcinomaGastric

carcinomabody

weightloss24718018415726512791055093103728685132»96

+ 235151523335*444240475550»479391988785»91

* Arithmetic mean of groups + standard deviation.

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MASON et al.—Cancer and Human Lirer Caialase 1481

TABLE 5

STATISTICALANALYTICALDATAOF HUMANLIVERCATALASEACTIVITYIN SOLUBLEFRACTION(SUPERNATANT)IN TWO-BY-TWOFACTORIALANALYSIS:CANCER-NOCANCERANDBODYWEIGHTLoss-No BODYWEIGHTLoss

A. DATA OF TOTALS

< 10% body weight loss> 10% body weight lossNo.

cancer620

4411,061Cancer6044781,082Total1,2249192,143

B. ANALYSISor VARIANCE

Row meansColumn meansInteractionSubtotalWithin

groups

TotalSum

ofsquares4,651

231394,81317,75022,563D.f.1

1131619Mean

square4,651

231391,109F

ratio4.2

0.020.10F

.90 (1, 16)= 3.07

cancers. The mechanism of toxohormone production and action remains a challenge in the humancancer problem.

REFERENCES1. ADAMS,D. H. Mouse Liver Catalase, the Antagonistic

Effect of Tumor Tissue upon the Hormonal ControlMechanisms. Brit. J. Cancer, 5:409-16, 1951.

2. . Sex Differences in Tissue Catalase Levels, andTheir Relation to the Catalase Depressing Action of Tumors. Ibid., 10:748-57, 1956.

3. . The Effect of Sarcoma 37 on the Intracellular Distribution of Mouse Liver Catalase. Ibid., 13:704-10, 1959.

4. APPLEMAN,D.; SKAVINSKI,E. R.; and STEIN,A. M. Catalase Studies on Normal and Cancerous Rats. Cancer Research, 10:498-505, 1950.

5. BEER, R. F., JR., and SIZER,I. W. A SpectrophotometricMethod Measuring the Breakdown of Hydrogen Peroxideby Catalase. J. Biol. Chem., 196:133-49, 1952.

6. BLUMENTHAL,F., and BRAUN,B. Die Katalasewirkung innormaler und in carcinomatoser Leber. Ztschr. Krebs-forsch., 8:436-40, 1910.

7. COLOWICK,S. P., and KAPLAN,N. O. Methods in Enzy-mology, 11:775-81, 1955.

8. DIXON,W. J., and MASSET,F. J. Introduction to Statistical Analysis, pp. 139-188, 119-124. 2d ed. New York:McGraw-Hill, 1957.

9. GREENFIELD,R. E., and PRICE,V. E. Liver Catalase. J. Biol. Chem., 220:607-18, 1956.

10. GREENSTEIN,J. P. Chemistry of the Tumor-bearing Host.Biochemistry of Cancer, pp. 518-40. New York: AcademicPress, Inc., 1954.

11. GREENSTEIN,J. P.; JENRELTE,W. V.; and WHITE,J. TheLiver Catalase Activity of Tumor-bearing Rats and theEffect of Extirpation of the Tumors., J. Biol. Chem., 141:327-28, 1941.

12. GUBLER,C. J. Enzyme Activities and Ion Metabolism inCopper and Iron Deficiencies. J. Biol. Chem., 224:533-46,1957.

13. KIYOTA,O. U.; KTJNIHIRO,M.; JUNZO,I.; YASTJMASS,A.;and TOSHIO,K. Liver Catalase Activity of Patients withGastric Cancer. Tohoku J. Exper. Med., 67:159-61, 1958.

14. NAKAOAWA,S. Studies on Catalase Inhibitors in LivingBodies. Medicine and Biology, 36(1): 27-31, 1955.

15. NAKAHARA,W., and FÃœKÃœOKA,F. A Toxic Cancer TissueConstituent as Evidenced by Its Effect on Liver CatalaseActivity. Jap. M. J., 1:271-77,1948.

16. PRICE,V. E., and GREENFIELD,R. E. Catalase Fractionsfrom Normal and Tumor-bearing Rats. J. Biol. Chem.,209:363-75, 1954.

17. VANPILSÃœM,J. F. Essential Amino Acid Deficiency andEnzyme Activity. Arch. Biochem. & Biophys., 68:42-53,1957.

18. VON EULER, H., and HELLER, L. Katalaseaktivität inLeberfraktionen normaler und sarkomtragender Ratten.F. Krebsforsch., 66:393-406, 1949.

19. WOODS,M. W. Polyvinylpyrrolidone as Adjunct in Centrifugal Separation and Enzymic Assay of Subcellular Components. Proc. Soc. Exper. Biol. & Med., 87:71-73,1954.

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1960;20:1474-1481. Cancer Res   Edward E. Mason, Ting-fong Chin, Yao W. Li, et al.   Cancer and Human Liver Catalase

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