Search

Hospital isolates of Enterobacter cloacae were analysed by polyacrylamide gel electrophoresis for enzyme polymorphism and the results were compared with established serotyping, phage typing and biotyping techniques. Initially, the diversity of electromorphs of 13 enzymes was determined on a representative set of 62 distinct strains. Two broad clusters of strains were found in the species, and analysis by serotype suggested a limited diversity within the most frequent O serotypes. A subset of three enzymes, lactate dehydrogenase, 6-phosphogluconate dehydrogenase, glutamate dehydrogenase and an unidentified marker, were selected and used to type groups of hospital isolates. There was good general agreement between the two systems, although the enzyme method failed to distinguish between some strains with the same serotype. This method provided useful epidemiological information and, in the absence of established typing systems, it is a practical approach to subdividing the species.

The relative mobilities of six enzymes from the trophozoites of five axenically-cultured isolates of Giardia from human, cat, and guinea pig hosts were compared by starch and polyacrylamide gel electrophoresis. The six enzymes compared were malate dehydrogenase (NAD+) (MDH) (EC 1.1.1.37), malate dehydrogenase (decarboxylating) (ME) (EC 1.1.1.40), hexokinase (EC 2.7.1.1), 6-phosphogluconate dehydrogenase (EC 1.1.1.44), glucose-6-phosphate dehydrogenase (G6P) (EC 1.1.1.49), and alpha-glycerophosphate dehydrogenase (EC 1.1.1.8). The latter three enzymes have not been previously reported in Giardia. On the basis of zymogram patterns, the five Giardia isolates were divided into three zymodemes. Zymodeme I comprised human-1/England, human-1/Bethesda, and cat-1/Portland, Zymodeme II the guinea pig-1/Portland isolate, and Zymodeme III the human-1/Portland isolate. These zymodemes were further substantiated when several physical and kinetic properties of three of the enzymes, MDH, ME, and G6P, were examined. Our results, in which Giardia isolated from different mammalian hosts share multiple isoenzymes, question the validity of the practice of assigning Giardia species names on the basis of the animal host from which the protozoan was obtained.

Enzymes of glucose metabolism were assayed in crude cell extracts of Frankia strains HFPArI3 and HFPCcI2 as well as in isolated vesicle clusters from Alnus rubra root nodules. Activities of the Embden-Meyerhof-Parnas pathway enzymes glucokinase, phosphofructokinase, and pyruvate kinase were found in Frankia strain HFPArI3 and glucokinase and pyruvate kinase were found in Frankia strain HFPCcI2 and in the vesicle clusters. An NADP+-linked glucose 6-phosphate dehydrogenase and an NAD-linked 6-phosphogluconate dehydrogenase were found in all of the extracts, although the role of these enzymes is unclear. No NADP+-linked 6-phosphogluconate dehydrogenase was found. Both dehydrogenases were inhibited by adenosine 5-triphosphate, and the apparent Km's for glucose 6-phosphate and 6-phosphogluconate were 6.86 X 10(-4) and 7.0 X 10(-5) M, respectively. In addition to the enzymes mentioned above, an NADP+-linked malic enzyme was detected in the pure cultures but not in the vesicle clusters. In contrast, however, the vesicle clusters had activity of an NAD-linked malic enzyme. The possibility that this enzyme resulted from contamination from plant mitochondria trapped in the vesicle clusters could not be discounted. None of the extracts showed activities of the Entner-Doudoroff enzymes or the gluconate metabolism enzymes gluconate dehydrogenase or gluconokinase. Propionate- versus trehalose-grown cultures of strain HFPArI3 showed similar activities of most enzymes except malic enzyme, which was higher in the cultures grown on the organic acid. Nitrogen-fixing cultures of strain HFPArI3 showed higher specific activities of glucose 6-phosphate and 6-phosphogluconate dehydrogenases and phosphofructokinase than ammonia-grown cultures.

Pulmonary alveolar macrophages (PAM), obtained by bronchoalveolar lavage from 47 individuals, reduced hexavalent chromium [Cr(VI)] and decreased its mutagenicity. Their specific activity--mostly mediated by cytosolic, enzyme-catalyzed mechanisms--was significantly higher than in corresponding preparations of mixed-cell populations from human peripheral lung parenchyma or bronchial tree, or from rat lung or liver. At equivalent number of PAM, Cr(VI) reduction, total protein, and some oxidoreductase activities were significantly increased in smokers. No appreciable variation could be detected between lung cancer and noncancer patients. In rats, the Cr(VI)-reducing activity of PAM preparations was induced by Aroclor 1254. Thus, alveolar macrophages provide crucial defense mechanisms not only by phagocytizing metals, but also by metabolically reducing Cr(VI). The epithelial-lining fluid (ELF) also displayed some Cr(VI) reduction. Together with already investigated metabolic processes occurring inside lung cells, these mechanisms are expected to determine thresholds in the pulmonary carcinogenicity of chromium.

The genetic relationships within and between stocks of Trypanosoma cruzi were studied by the in vitro isolation of clones and sub-clones and by the comparison of their isoenzyme patterns in thin-layer starch-gel electrophoresis. Altogether 13 clones and 36 sub-clones were isolated from stocks CL89 and Y207 of T. cruzi. Employing the enzymes L-alanine aminotransferase (ALAT), L-aspartate aminotransferase (ASAT), glucose phosphate isomerase (GPI), malic enzyme (ME), malate dehydrogenase (MDH), glucose-6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), and phosphoglucomutase (PGM), two zymodemes, B and C, emerged among the clones with distinct banding patterns. These zymodemes were consistently distinguished by ALAT, ASAT, GPI, G6PD, 6PGD, and PGM and the differences in enzyme profiles were simultaneously reflected in all six enzyme systems. That the enzymic characters as genetic determinants are stable was demonstrated after recloning and successive replica-platings, i.e., the distinct enzyme patterns remained consistent and homogeneous, the siblings retained the enzyme profiles of their parental clones, and no segregation of the enzyme patterns was observed. Our data from clone and sub-clone examinations show that the isoenzymes act as labels to characterize T. cruzi stocks. Furthermore, enzyme variation was demonstrated among clones isolated from stock CL89.

Enzymatic study in noncultured amniotic fluid cells from 87 women at different stages of pregnancy was performed by electrophoresis. Of these patients, 33 had normal pregnancies and the remaining 54 had various complications of pregnancy. The electrophoretic patterns of lactate dehydrogenase, alpha-esterase, and acid phophatase were altered in samples obtained either at different gestation periods or from normal and complicated pregnancies. Malate dehydrogenase and protein bands did not show any alteration. Glucose-6-phosphate dehydrogenase and alkaline phophatase were not detectable. 6-Phosphogluconate dehydrogenase was detectable in samples of complicated pregnancies. The significance of the altered patterns of enzymes is discussed.

This study is concerned with the histochemical activity of hydrosteroid dehydrogenases (HSD), glucose-6-phosphate dehydrogenase (G6PDH), and 6-phosphogluconate dehydrogenase (6PGDH) in testis and epididymis of 20 male Pekin ducks. Various 3 beta-HSD, the G6PDH, and the 6PGDH were localized in these organs. 3 beta-etiocholane was much quicker utilized by the 3 beta-HSD than epiandrosterone, dehydroepiandrosterone, and androstendiol. The capability of testis Leydig cells of utilizing the 3 beta-etiocholane was higher in winter than in summer. The G6PDH and the 6PGDH in the Leydig cells and basal cell layer of the tubuli seminiferi contorti were active throughout the year. There was weak 3 beta-HSD activity in the epithelia of the ductuli efferentes distales and in the ductus epididymidis from March to June. The G6PDH and the 6PGDH showed positive reaction in the rete testis and ductuli efferentes proximales throughout the year. The 3 beta-HSD activity in testis and epididymis is saisonally different: In summer, there is no 3 beta-HSD activity in both organs. In winter, the 3 beta-HSD is demonstrable in the testis but not in the epididymis. In spring, there is 3 beta-HSD activity in the epididymis but not in the testis.

We have investigated the occurrence of dosage compensation in D. willistoni and D. pseudoobscura, two species whose X chromosome is metacentric with one arm homologous to the X and the other homologous to the left arm of chromosome 3 of D. melanogaster. Crude extracts were assayed for isocitrate dehydrogenase (XR), glucose-6-phosphate dehydrogenase (XL?), 6-phosphogluconate dehydrogenase (XL?), and alpha-glycerophosphate dehydrogenase (chromosome 2) in D. willistoni, and for esterase-5 (XR), glucose-6-phosphate dehydrogenase (XL?), 6-phosphogluconate dehydrogenase (XL?) and amylase (chromosome 3) in D. pseudoobscura. Our results indicate that a mechanism for dosage compensation is operative in both arms of the X chromosome of these two species.

The antigenic architecture of membrane vesicles prepared from Escherichia coli ML 308--225 has been studied using crossed immunoelectrophoresis. Progressive immunoadsorption experiments conducted with control vesicles and with physically disrupted vesicles were used to monitor and quantitate the expression of 14 different immunogens. Eleven immunogens, including NADH dehydrogenase (EC 1.6.33.3), D-lactate dehydrogenase (EC 1.1.1.27), dihydro-orotate dehydrogenase (EC 1.3.3.1), 6-phosphogluconate dehydrogenase (EC 1.1.1.43), polynucleotide phosphorylase (EC 2.3.7.8), and beta-galactosidase (EC 3.2.1.23), exhibit minimal expression (10% or less) unless the vesicles are disrupted. Three unidentified antigens are expressed to a similar extent in untreated and disrupted vesicles. Consideration of these and other results [Owen, P., & Kaback, H. R. (1978) Proc. Natl. Acad. Sci. U.S.A. 75, 3148] in terms of membrane polarity, dislocation of antigens, and possible transmembrane orientation of some immunogens reveals that over 95% of the membrane in the vesicle preparations is in the form of sealed sacculi with the same orientation as the intact cell. Furthermore, antigens are distributed across the membrane in a highly asymmetric manner, indicating that dislocation of components from the inner to the outer surface of the membrane during vesicle preparation does not occur to an extent exceeding 10%.

Phosphogluconate dehydrogenase (PGD) and xanthine dehydrogenase (XDH) were genetically studied by agar gel electrophoresis in the mosquito Aedes togoi. Adult homogenates displayed banding patterns with one zone of activity of either enzyme. Eight backcrosses were conducted to map the two loci, Pgd and Xdh, to linkage group III with the following arrangement: Pgd--(ca. 15 map units)--bl (bleached pupa)--(ca.5)--y (yellow larva)--(38.0 +/- 1.7)--pm (plum eye)--(6.1 +/- 0.9)--Xdh. All Pgd and Xdh loci thus far mapped in other mosquito species are reviewed for a comparison with those in this species.

A case of homicide with postmortem amputation of the penis was described. The penis found later in a home freezer in the apartment of the culprit showed "zigsaw" fit to the victim. In order to substantiate the morphological findings, phenotypes of three isozymes, phosphoglucomutase1 (PGM1), phosphogluconate dehydrogenase (PGD), and esterase D (EsD) together with ABO blood type were determined in the skin of the penis amputated. All of the four genetic markers determined with the penis were the same as those determined with heart blood of the victim.

After administration of insulin into rats activity of dehydrogenases of pentosephosphate pathway was not altered in cortex and medulla of kidney as distinct from the liver tissue. Concentration of free fatty acids was slightly lower in soluble fraction of kidney cortex than in blood plasma and higher than in preparation from kidney medulla. Alterations in content of free fatty acids were not observed in the preparations of kidney of rats with alloxan diabetes.

Sixty isolates of Plasmodium falciparum collected from hospital in-patients in Rangoon and out-patients from villages near Rangoon were tested in vitro for chloroquine sensitivity and then cultured to carry out starch gel electrophoresis of the following parasite enzyme: glucose phosphate isomerase (GPI) (EC.5.3.1.9.), NADP-dependent glutamate dehydrogenase (GDH) EC.1.4.1.4), lactate dehydrogenase (LDH) (EC.1.1.1.27) and 6-phosphogluconate dehydrogenase (6PGD) (E.C.1.1.1.44). Variations were observed in three (GPI, GDH and LDH) of the four enzymes examined. Three forms of parasite GPI, two forms of parasite GDH, two forms of parasite LDH and only one form of parasite 6PGD were identified. Enzyme polymorphism was more common in isolates with higher resistance to chloroquine. One isolate with parasite GDH-2 and six isolates with parasite LDH-2 were recorded from chloroquine-resistant cases.

The steroid hormone has an important role in the early stages of reproduction. There has been abundant histochemical evidence that oocytes contain steroid hormones and are able to synthesize these hormones. But there have been few methods of analyzing one oocyte biochemically because it is too small and light. In order to study steroidogenesis in the oocyte, a microassay method sensitive enough to analyze the enzyme activities in one oocyte was developed using enzymatic cyling for amplifying the reaction product to 10,000-fold. An oil-well technique and a microtube method were applied in the assay for achieving the reaction in a medium as small as 1.0 to 5.0 microliters under a stereomicroscope. Immature Wistar rats were superovulated by PMS-hCG administration. Oocytes were collected by puncturing the follicle and flushing the tube. They were freeze-dried after washing to remove cumulus cells. The dry weight of one oocyte was 51.2 +/- 6.2 ng in a quartz fiber fishpole balance. The activity of 3 beta hydroxysteroid dehydrogenase (3 beta HSD) (picomol/oocyte/hr, substrate:pregnenolone) in the PMS-treated oocyte was 2.66 +/- 0.59, which corresponds to 3 times the activity of the ovarian homogenate as control, indicating the high capacity of oocytes to produce progesterone. The activity increased significantly (P less than 0.01) by hCG administration up to 4.17 +/- 0.29 after ovulation, suggesting that gonadotropin regulates steroidogenesis in the oocyte. The activities of G6PD and 6PGD were 8.41 +/- 1.09 picomol/oocyte/min and 3.85 +/- 2.02 picomol/oocyte/hr, respectively. The high activity of G6PD (more than 10 times that of the ovarian homogenate) suggests that the pentose phosphate shunt concerned with steroidogenesis is active in the oocyte. HCG decreased the activities of both G6PD and 6PGD. The present results show that steroidogenesis in the oocyte is very active under the control of gonadotropin, suggesting that steroid hormones may play an important role in oocyte maturation, ovulation and fertilization.

The glucose flow in Xanthomonas campestris was investigated with radio-labelled glucose and by enzymological studies. Only 7% of the radioactivity was incorporated into the cell material, but 41% was oxidized to carbon dioxide and 28% transformed to xanthan. Up to 16% of cell dry weight consisted of the polysaccharide glycogen. In the presence of 2.7 mM methionine, which is an inhibitor of xanthan formation, increased carbon dioxide formation (51%) occurred. This increase was in accordance with a twofold increase in the NAD-dependent isocitrate dehydrogenase activity. The other carbon dioxide liberating enzyme, 6-P-gluconate dehydrogenase, was not influenced by methionine, but its occurrence indicates the presence of an active pentose phosphate pathway in X. campestris. Among the other enzymes detected in X. campestris was glucose dehydrogenase. The presence of this enzyme together with hexokinase indicates the operation of two different glucose metabolizing steps: one oxidative, the other phosphorylative. Only the latter directly provides phosphorylated glucose as a precursor for the activated sugars required for xanthan synthesis.

A fluorimetric procedure for the determination of phosphoglycerate kinase in single human hair follicles is described. Enzyme studies on different parts of hair follicles after dissection show that the distribution of glucose-6-phosphate dehydrogenase matches that of phosphoglycerate kinase. Glucose-6-phosphate dehydrogenase can therefore be used as a reference enzyme to compensate for differences in hair follicle sizes. It was shown that the variation in the values found in individual hair follicles is improved by relating phosphoglycerate kinase to glucose-6-phosphate dehydrogenase activity. In areas of the world where glucose-6-phosphate dehydrogenase deficiency occurs frequently, an autosomally inherited reference enzyme may be preferred. It is shown that 6-phosphogluconate dehydrogenase is useful in this respect. Upon storage a gradual drop in the activity of all three enzymes was observed, but the rate of decrease was about equal: the enzyme activity ratio was, therefore, almost unaffected for a period of one week. This allows the determination of phosphoglycerate kinase even in mailed hair follicles.

A man aged 42 years died from a hemorrhage resulting from stab wounds in his neck, chest and abdomen. The MN blood group and PGM1 phenotype of the victim's heart blood were inconsistent with those of the blood stains around the victim. The victim was transfused with 4000 ml of blood during the 9 hours before his death. Because PGM1 phenotypes obtained from blood stains at the scene of the crime and from the weapon corresponded with those from the victim's dental pulp, peripheral nerve and bone marrow, the victim's blood groups and isoenzymic phenotypes could be determined accurately.

At Makthlawaiya, in the Paraguayan Chaco, the prevalence of Trypanosoma (Schizotrypanum) cruzi infection among both domestic Triatoma infestans and domestic dogs was 38%, and IgG anti-T. cruzi antibody was detected by the quantitative enzyme-linked immunosorbent assay (ELISA) in 80% (105/133) of human sera. Ninety percent (25/28) of T. cruzi strains isolated from both T. infestans and dogs showed heterozygous isoenzyme profiles for glucose phosphate isomerase, phosphoglucomutase and 6-phosphogluconate dehydrogenase. These strains appeared to be closely related to Bolivian zymodeme 2. Three Paraguayan T. cruzi strains showed homozygous isoenzyme profiles, similar to those of major Brazilian zymodemes. It was concluded that T. cruzi strains with heterozygous isoenzyme profiles predominate in domestic transmission cycles in this highly endemic area of the Paraguayan Chaco.

Perfluorodecanoic acid (PFDA) alters the circulating level of thyroid hormones, but the physiological significance of this change at the target tissue remains to be defined. To this end, the activities of thyroid-responsive hepatic enzymes were examined in adult male rats 1 week after treatment with a single dose of PFDA (20, 40 or 80 mg/kg). Since PFDA treatment caused a dose-related reduction in feed intake, vehicle-treated rats pair-fed to their counterparts receiving PFDA were used to determine if any of the PFDA-induced alterations in enzyme activity were secondary to hypophagia. Following the administration of PFDA, L-glycerol-3-phosphate dehydrogenase, a liver mitochondrial enzyme sensitive to thyroid status, exhibited a modest increase in activity, whereas that of succinate dehydrogenase, a constitutive mitochondrial marker enzyme, was similar in both PFDA-treated rats and their pair-fed counterparts at all dose levels examined. Activity of cytosolic lactate dehydrogenase was also augmented modestly in livers of rats receiving PFDA. In contrast, activity of cytosolic malic enzyme, a thyroid-responsive enzyme, was increased markedly in PFDA-treated rats. Hepatic activity of glucose-6-phosphate dehydrogenase, which also responds to alterations in thyroid status, exhibited a modest increase with 20 and 40 mg/kg PFDA but was similar in both PFDA-treated rats and their pair-fed counterparts at the 80 mg/kg dose level. Absolute and relative liver mass was elevated in PFDA-treated rats at all dose levels in comparison to the appropriate vehicle-treated pair-fed animals. Total hepatic content of DNA was maintained in PFDA-treated rats at all dose levels, whereas a significant decrease in liver DNA was found in the vehicle-treated rats pair-fed to animals receiving 80 mg/kg PFDA. Following administration of PFDA, protein content per total liver was similar to that of their pair-fed counterparts. Thus, the pattern of activity of thyroid-responsive hepatic enzymes was not compatible with a functional shift toward a lessened thyroid status in rats treated with PFDA.

During aging, there is a decrease in the activity of the 6-phosphogluconate dehydrogenase enzyme in rat liver. The "old" 6-phosphogluconate dehydrogenase enzyme is about 26% less active than the "young" enzyme. In this paper, some biochemical and chemical properties of this enzyme are studied. 2,4,6-Trinitro-benzenesulfonic acid measurements indicate that the old enzyme has 11 lysine residues less than the young enzyme. The proteolysis with trypsin produces more peptides in the young enzyme than in the old one. However, similar numbers of peptides were produced when endoproteinase Arg-C was used on both enzymes. Moreover, the treatment of the young enzyme with ascorbate for 15 min produces the loss of 8 lysine residues. These results suggest that during aging there is a modification of the lysine residue, and this could be involved in the loss of its enzymatic activity.

The activities of six enzymes associated with carbohydrate metabolism were measured in 24 benign prostatic hyperplasias and 29 carcinomas of the prostate with the intention of investigating whether these measurements might eventually predict the clinical course of the disease. The following findings are recorded: the activities of PFK, alpha-GPDH and PHI were significantly lower in carcinomas. Comparison between well and poorly-differentiated carcinomas indicated a significantly lower activity of 6PGDH and higher alpha-GPDH/6PGDH ratios in the latter. These findings are the opposite of those observed in carcinomas of the breast.

The electrophoretic resolution of seven relatively polymorphic human gene-enzyme systems expressed in tissue culture cells can be used as a sensitive genetic monitor for intraspecific cell contamination. An identical genotype at each of the same allozyme loci provides a 95% (or greater) confindence estimate of the identity of two cultured lines, on the basis of the allelic frequencies of the seven enzyme loci in natural populations and in populations of independently derived cultured cells. Of 27 commonly used human cell lines examined, only one of 351 pairwise comparisons proved genetically indistinguishable.

In this study, isozyme patterns for 14 different enzymes were compared for culture strains of Leishmania braziliensis, L. hertigi, L. mexicana, L. donovani, L. tropica, and L. adleri. The isozyme separation was made by means of cellulose acetate electrophoresis. Each of the species had distinct isozyme patterns for aspartate aminotransferase, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and fructokinase. For other enzymes, two or more species had identically migrating bands; however, by using combinations of the other 10 enzymes it was possible to separate any one of the six species. In addition to these interspecific differences the Panama strains of L. braziliensis had two different malic dehydrogenase isozyme patterns; therefore, they fell into two distinct groups. These strains otherwise had identical isozyme patterns.

The effects of administration of opiates and naloxone on the activities of PFK, 6PGDH and alpha-GPDH and alpha-GPDH/6PGDH ratios in human breast carcinomas were investigated in patients awaiting mastectomy. Injection of naloxone or fentanyl into an antecubital vein resulted in a statistically significant reduction in the activity of alpha-GPDH. Fentanyl was also effective in reducing the activity of 6PGDH. Injection of morphine into a branch of the internal mammary artery during mastectomy failed to induce changes in the activities of any of the enzymes but injection of naloxone resulted in a significant rise in the activity of 6PGDH. It is postulated that these alterations in the activities might not be associated with the binding of these drugs to opiate receptor proteins in the carcinoma. Furthermore opiate agonists or antagonists might not produce the required changes in the activities of any of the enzymes.

Membrane vesicles isolated from Escherichia coli ML 308--225 have been analyzed by crossed immunoelectrophoresis, and immunoprecipitates corresponding to the following cellular components have been identified: ATPase (EC 3.6.1,3), two or three NADH dehydrogenases (EC 1.6.99.3), D-lactate dehydrogenase (EC 1.1.1.27), glutamate dehydrogenase (EC 1.4.1.4), dihydro-orotate dehydrogenase (EC 1.3.3.1), 6-phosphogluconate dehydrogenase (EC 1.1.1.43), polynucleotide phosphorylase (EC 2.3.7.8), beta-galactosidase (EC 3.2.1.23), lipopolysaccharide, and Braun's lipoprotein. The cellular origin of many of the vesicle immunogens is determined, and Braun's lipoprotein is used as a marker to quantitate the extent of outer membrane contamination (less than 3%). Membrane antigens are also characterized with regard to their amphiphilic or hydrophilic properties by charge-shift crossed immunoelectrophoresis. Furthermore, the following immunogens cross-react with components in membrane vesicles prepared from Salmonella typhimurium: one of the three NADH dehydrogenases, ATPase, polynucleotide phosphorylase, 6-phosphogluconate dehydrogenase, Braun's lipoprotein, and three unidentified antigens. In the accompanying paper [Owen, P., & Kaback, H. R. (1979) Biochemistry 18 (following paper in this issue)] quantitative immunoadsorption is utilized to establish the topology of the vesicles with respect to the distribution of antigens on the inner and outer faces of the membrane.

The studies conducted showed that the beginning of immunogenesis following the administration of live plague vaccine was preceded by a period of primary toxic action of multiplying microbes when the activity of a number of enzymes in the organs of guinea-pigs was temporarily reduced. Subsequently, inductive and productive phases of formation of antibodies started against the background of normalization and growth of the enzymatic activity associated with mobilization of energy. The earliest hyperplastic changes during immunogenesis occurred in the thymus-dependent areas of the lymphoid organs. Somewhat later, hyperplasia of B-structures dependent on the bone marrow started to prevail.

At each of five loci in 829 Escherichia coli clones from 156 samples from diverse natural sources, electrophoretic analysis reveals a prominent mobility class (frequency over 0.70) and 2 to 11 distinct mobility classes at lower frequencies. The frequency distribution of the classes argues against the importance of neutral mutations in allozymic variation. Heterosis is not the universal cause of genic polymorphism.

The potential therapeutic effects of differentiating agents on leukemic and solid tumor cells are being evaluated worldwide. These effects can be followed by morphologic as well as biochemical parameters. The enzymatic profile of four enzymes and the level of carcinoembryonic antigen were studied in 24 human colorectal carcinoma specimens and their adjacent uninvolved mucosa. The enzymes studied were thymidine kinase and 6-phosphogluconate dehydrogenase as markers of proliferation, and alkaline phosphatase and gamma-glutamyl transpeptidase as markers of differentiation. A consistent finding was a marked increase in the activities of thymidine kinase and 6-phosphogluconate dehydrogenase in the tumor cells as compared with the adjacent normal mucosa. The activity of gamma-glutamyl transpeptidase was not significantly different between tumor and uninvolved colon tissue. Alkaline phosphatase activity was markedly reduced in the tumor specimens. A relationship between the degree of differentiation and the degree of penetration and CEA expression was demonstrated in the tumor specimens as well as in their surrounding uninvolved mucosa.

A versatile fluorimetric assay based on the reduction of resazurin to resorufin demonstrated high specific activities for a number of important pyridine nucleotide-linked dehydrogenases in tobacco leaves. The Michaelis constant for the important photosynthetic enzyme, D-glyceraldehyde-3-phosphate:NADP+ oxidoreductase (EC 1.2.1.13), determined by the fluorimetric method, was considerably lower than constants determined by conventional extraction and assay methods reported for the enzyme from other plants. The sensitivity of the fluorimetric method enabled the use of dilute enzyme preparations with resultant low background and high substrate specificity. Inclusion of the anti-oxidant diethyldithiocarbamate in the extraction medium preserved the enzymes during extraction. Primary amines inhibited competitively, and phenazine methosulfate non-competitively each of the eight dehydrogenases tested with the fluorimetric assay. The Mn2+ dependence of NADP-linked dehydrogenases specific for isocitrate and malate was confirmed. The method is rapid, requires a simple combination of ingredients and should be useful for surveying dehydrogenase activity in leaves.

In an experimental population of the japanese quail Coturnix coturnix japonica, two polymorphic ovoglobulin loci, two polymorphic esterase loci, and PGD versus G-6-PD loci are in epistatic disequilibrium. Epistatic interactions occur between structurally and physiologically very closely related proteins and enzymes.

The enzymes of 35 adult Paragonimus uterobilateralis were analysed using thin-layer starch gel electrophoresis. From a total of 21 enzyme systems studied, 15 proved to be useful for the description and recognition of this species. All individuals were identical concerning 11 enzymes. In four remaining enzymes, alanine aminotransferase (ALAT, hexokinase, aspartate aminotransferase and phosphogluconate dehydrogenase, two or three variants, also being partly typical for this species were observed. In a comparison involving seven different enzymes, there were no differences between the electrophoretic patterns of 35 adult and 24 juvenile P. uterobilateralis. Additional examinations of 30 adult P. uterobilateralis with isoelectric focusing on ultrathin-layer polyacrylamide gels revealed clearer separations of enzymes. The method showed corresponding results or identiy of all individuals tested with three representative enzymes (ALAT, glucosephosphate isomerase and adenylate kinase).

The reservoir animals, sandfly vectors and strains of Leishmania from foci in the southern region of Israel were studied. The rodent host species are: Psammomys obesus, Meriones crassus and probably Nesokia indica. The vector species are Phlebotomus papatasi, which were caught at all collecting sites and Ph. sergenti, which were collected in the area of the Dead Sea and in the Central Arava. Strains of Leishmania major isolated from rodents, vectors and man were serologically and enzymologically identical with regard to their excreted factor (EF) serotypes, their malate dehydrogenase (MDH), glucose-phosphate isomerase (GPI) and glucose-6-phosphate dehydrogenase (G6PDH) enzyme variant types, but exhibited three variant subtypes of 6-phosphogluconate dehydrogenase (6PGDH). The distribution of the 6PGDH subtypes correlates with three different geographical locations. Scarcity of water is the main factor limiting the biotopes of the sandflies and the spread of leishmaniasis. The subjects discussed are the dependence of sandfly distribution on rodent-burrow depth in arid areas and the inter-relationship between the leishmanial subtypes, vectors and hosts.

Gonadectomized male and female rats were treated with equimolar doses of estradiol benzoate (EB) or testosterone pripionate (TP) daily for one week and enzyme activities were measured in the basomedial hypothalamus, corticomedial amygdala, and pituitary. In females, the hypothalamus showed estrogen-dependent increases in glucose-6-phosphate dehydrogenase (G6PDH), isocitrate dehydrogenase (ICDH), and malate dehydrogenase (MDH). Activities of ICDH and MDH were elevated in the amygdala. In the pituitary, estrogen administration resulted in increased levels of G6PDH, 6-phosphogluconate dehydrogenase (6PGDH), and lactic dehydrogenase (LDH). The estrogen antagonist, MER-25, effectively blocked estrogen-dependent increases in pituitary G6PDH and 6PGDH. Administration of TP did not result in changed enzyme levels. In males, treatment with EB and TP resulted in significant elevations in some but not all enzymes that were increased by EB in the female. Estrogen-dependent increases of activity in males were noted in pituitary G6PDH, 6PGDH, and LDH, in hypothalamic MDH, and in amygdaloid ICDH. Administration of TP led to increased levels of pituitary G6PDH, 6PGDH, LDH, ICDH, and MDH, hypothalamic ICDH and G6PDH, and amygdaloid MDH. The pattern of enzyme changes found in male and female brain and pituitary is discussed in relation to behavioral responses to gonadal hormones, nuclear uptake of gonadal hormones, and metabolism of androgen.

The Griqua of Campbell in the Cape Province of South Africa are reputed to be descended from an amalgam of Khoi ("Hottentots") with various Caucasoid, Negroid and East Asian elements at the Cape of Good Hope. A stormy history of migration and deprivation has left this small residuum on the edge of the Kalahari Desert. It was hoped that a sero-genetic study of these people, who proudly preserve their identity, might provide further insights into the genetic consitution of their Khoi forebears. Despite the retention of the language of their own, an social mores distinct from of other Mixed populations, they have been shown, however, to have received an appreciable inflow of non-Khoisan genes. This has probably resulted from social factors such as the prestige attaching to Griqua membership and the willingness of the Griqua to assimilate outsiders, combined with the tendency of Campbell Griqua to migrate elsewhere. There are no significant differences between "Griqua" members of the population and those who are not recognized as "Griqua."

A method of mapping genes which specify enzymes without the necessity of obtaining genetic variants has been explored. Three enzymes whose structural genes have known genetic positions were chosen to see if the relationship between gene dosage and enzyme activity could be used as a tool in cytological localization. Zw, the gene specifying G6PD, is located in the X chromosome region, 18D-18F. The structural gene for 6PGD, Pgd, maps in the X chromosome bands 2C1-2E1. Idh-NADP, the gene which specifies IDH-NADP, is found on the third chromosome, in bands 66B-67C.

The value of 6-phosphogluconate dehydrogenase (6-PGDH) activity as a cytosolic marker of tumour cellularity has been assessed, together with its use as a prognostic indicator for primary breast cancer in 87 patients over 4 years. 6-PGDH activity shows a good correlation with histologically-assessed tumour cellularity in a sample of 114 patients (correlation coefficient = 0.83). Patients whose primary breast tumour had a high 6-PGDH activity showed poor relapse-free survival times when compared to those with low 6-PGDH activities (Log Rank chi 2 = 6.87, P less than 0.01). This compared with a Log Rank chi 2 of 2.22, P less than 0.20 for oestrogen positive and negative patients. These results suggest that 6-PGDH activity is a better prognostic indicator in primary breast cancer than oestrogen receptor status.