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PubMed1990

Differentiation of clinical isolates of Entamoeba histolytica by using specific DNA probes.

Most individuals infected with Entamoeba histolytica are reported to be clinically asymptomatic. On the basis of the electrophoretic migration of hexokinase and phosphoglucomutase isoenzymes, two groups of E. histolytica isolates have been classified. Those derived from symptomatic cases were found to have fast-migrating hexokinase bands and were labeled pathogenic. The others, isolated from cyst passers, had (in most cases) slow-migrating bands and were called nonpathogenic. Differences between these two groups of E. histolytica were found recently at the DNA level. Two sets of different DNA probes derived from tandemly repeated sequences present in extrachromosomal circular DNA elements in each group of E. histolytica were characterized. Using these probes with procedures for direct hybridization of trophozoites on nylon membranes, we could correctly correlate hexokinase electromobility with the DNA hybridization signal of 81 different isolates of E. histolytica. The advantages of using DNA probes lie in their sensitivity (fewer than 200 trophozoites can be detected) and specificity. The probes hybridized only with amebae from the E. histolytica species and not with other enteric protozoa and can be useful as a diagnostic tool.

Animals↗
PubMed1979

Genetic polymorphisms in diabetics and non-diabetics.

Phenotype distributions of some genetic polymorphisms are reported in a sample of 721 diabetics and 515 non-diabetic, non-blood donor controls. Reference is also made, in the case of the ABO and Rhesus systems, to previously published results for blood donors resident in the Durham area. Non-insulin-taking diabetics show an increased frequency of blood group A1 (and A1 + A2) when compared with controls. This difference is particularly marked in male diabetics. When diabetics are compared with age matched controls, the difference is confined to the older cases. It is proposed that this effect is predominantly the result of a deficiency of group A1 in controls rather than the result of increased susceptibility to the disease among A1 people. No association with any of the Rhesus phenotypes is shown. In non-diabetics, the results suggest an enhanced survival value for the rr genotype. No significant associations are seen when the MNSs, Kell, Lewis, Duffy, haptoglobin, red cell acid phosphatase, phosphoglucomutase, adenylate kinase, and adenosine deaminase distributions in these groups of subjects are compared.-

ABO Blood-Group System↗
PubMed1982

Isoenzyme characterisation of Trichomonas vaginalis.

Clones of 32 strains of Trichomonas vaginalis isolated from patients attending a venereal diseases clinic were compared among themselves and with authentic Pentatrichomonas hominis on the basis of their isoenzyme patterns for eight enzymes by thin-layer starch-gel electrophoresis. The enzymes examined were: glucose phosphate isomerase (GPI); phosphoglucomutase (PGM); malic enzyme (NADP+) (ME); hexokinase (HK); malate dehydrogenase (NAD+) (MDH); glucose-6-phosphate dehydrogenase (G6PD); aldolase (ALD); and lactate dehydrogenase (LDH). From the isoenzyme patterns of four enzymes (LDH, MDH, HK, and GPI) the strains of T vaginalis could be divided clearly into five groups. PGM showed differences in only one strain, while two other enzyme patterns (ME and ALD) were the same for all the strains of T vaginalis tested. All isolates were clearly distinguishable from P hominis. Although G6PD patterns were not sharp some differences were evident among T vaginalis strains.

Animals↗
PubMed1986

Chronic stimulation of mammalian muscle: changes in enzymes of six metabolic pathways.

Twenty-one enzymes of different metabolic systems were measured in the rabbit fast-twitch tibialis anterior (TA) muscle after electrical stimulation (10 Hz, 24 h/day) for 1 day to 10 wk. Nine analytical methods are either new, (3-oxoacid CoA-transferase, branched-chain-amino-acid aminotransferase, carnitine acetyltransferase, thiolase), improved (glutamate dehydrogenase, glycogen synthase, adenylic acid deaminase), or specially adapted (hexokinase, phosphoglucomutase). The activities (based on protein) of 12 mitochondrial or partly mitochondrial enzymes were lower in control TA than in control (slow) soleus (30-84% of soleus level). After 2 wk, 11 of these had surpassed the control soleus level. Maximal increases (3- to 14-fold) occurred after 2-5 wk, and thereafter six of the enzymes declined, whereas the other five maintained or increased their levels. Five glycolytic and two high-energy phosphate transfer enzymes, originally much higher in control TA than in control soleus, decreased gradually to levels at 8-10 wk only 27-123% higher than in soleus. Noncollagen protein concentration dropped 46%, explained largely by a sixfold increase in extracellular (chloride) space and a modest increase in collagen. The data constitute strong evidence for coordinate regulation of (mainly cytosolic) enzymes of glycolysis, glycogenolysis, gluconeogenesis, and high-energy phosphate transfer. Changes in the (mainly mitochondrial) enzymes of oxidative metabolism were more divergent, partly because of a hitherto undescribed secondary phase in the metabolic response. This phase may reflect a lower energy consumption in muscles adapted to continuous activity.

Adaptation, Physiological↗
PubMed1988

G-1,6-P2 in human skeletal muscle after isometric contraction.

The content of glucose 1,6-bisphosphate (G-1,6-P2), an in vitro activator of phosphofructokinase (a rate-limiting enzyme for glycolysis), and the glycolytic rate in skeletal muscle during isometric contraction have been determined. Subjects contracted the knee extensor muscles at two-thirds maximal voluntary force to fatigue. Biopsies from the quadriceps femoris muscle were obtained before and immediately after contraction. G-1,6-P2 increased in all subjects from a mean of 101 +/- 15 (SE) mumol/kg dry wt at rest to 128 +/- 24 at fatigue (P less than 0.05). Muscle glucose did not change significantly, whereas hexosemonophosphates were significantly increased after contraction. The glycogenolytic and glycolytic rate averaged 70.0 +/- 13.8 and 47.3 +/- 6.7 mmol.kg dry wt-1.min-1, respectively, and the glycolytic rate was positively correlated with the accumulation rates of fructose 6-phosphate (F-6-P) (r = 0.95, P less than 0.01) and G-6-P (r = 0.96, P less than 0.01). Phosphocreatine and ATP decreased by 87 and 17%, respectively, whereas ADP increased by 31% after contraction. These data demonstrate that intense, short-term isometric contraction results in an elevation of the muscle content of G-1,6-P2. The increase in G-1,6-P2 could not be accounted for by the side reactions of phosphoglucomutase or phosphofructokinase. It remains to be determined whether the observed increase in G-1,6-P2 is sufficient to account for the high glycolytic rate during intense exercise. The lack of increase in muscle glucose while G-6-P increased (which will inhibit hexokinase) suggests that the debranching enzyme complex was not active during contraction.

Adenine Nucleotides↗
PubMed1985

Enzymes and metabolites of glycogen metabolism in canine cardiac Purkinje fibers.

Canine Purkinje fibers were isolated by microdissection and analyzed for four enzymes of glycogen metabolism and eight related metabolites. Purkinje fiber glycogen levels were very high, confirming earlier reports. Glycogen synthesizing enzymes, glycogen synthase and UDP glucose pyrophosphorylase, were on the average 47 and 70% higher, respectively, in Purkinje fibers than in myocardium. Phosphorylase activity was approximately equal in the two tissue types, and phosphoglucomutase was 31% lower in Purkinje fibers. The metabolites of glycogen 6-phosphate were all higher in Purkinje fibers (P less than 0.001), but glucose 1,6-bisphosphate was lower by 50%. Phosphocreatine and ATP remained high in Purkinje fibers during 2 min of ischemia, while the phosphocreatine level in myocardium was falling by 75%. The results of this study suggest that the high glycogen synthetic capability, high precursor levels, and overall lower metabolic rate in Purkinje fibers compared with myocardium may explain the much higher glycogen levels.

Adenosine Triphosphate↗
PubMed1984

Subcellular sites of insulin hydrolysis in renal proximal tubules.

The subcellular sites of insulin degradation as measured by trichloroacetic acid precipitation were defined for rabbit renal proximal tubule cells. Fractionation in linear sucrose gradients of the postnuclear supernates prepared from isolated proximal tubule segments revealed three pools of insulin hydrolytic activity. Insulin hydrolytic activity assayed at pH 3.5 distributed in the gradients in a manner nearly identical to the activity of the lysosomal enzymes, N-acetyl-beta-glucosaminidase and alpha-mannosidase. At pH 7.4 the insulin-degrading activity distributed in a bimodal fashion with the major component following the cytosolic enzyme, phosphoglucomutase, and the minor component nearly identically overlapping with the activity of the inner mitochondrial enzyme, cytochrome oxidase. Upon microperfusion of 125I-insulin through proximal straight nephron segments, metabolites of the hormone were not observed in the collected perfusates for six of eight experiments. Average values for percent intact insulin in the original and collected perfusates showed no significant difference. These data suggest that three potential sites for insulin hydrolysis are present in proximal tubule cells, including lysosomes, the cytosol, and mitochondria. The results do not support the concept of degradation occurring at the brush border or contraluminal membranes.

Animals↗
PubMed1995

Contractility and phenotype transitions in serosal thickening of obstructed rabbit bladder.

Partial outlet obstruction of rabbit bladder induces serosal thickening and smooth muscle (SM) cell hypertrophy that are accompanied by phenotypic changes in the expression of cytoskeletal and cytocontractile proteins. In the present study, we compare the observed progressive phenotypic changes with the contractile responses of strips of the thickened serosa. At 15 days after partial outlet obstruction, although cells in thickened serosa demonstrate the presence of nonmuscle (NM) myosin of A-like type, vimentin, and SM alpha-actin, no contractile responses of this tissue were noted. At later times (30 days), this tissue expressed in addition SM myosin, and this pattern was paralleled by the development of KCl-stimulated contractility. It is only after 60 days that the serosa demonstrated the expression of desmin, phosphoglucomutase (PGM)-related protein, and was locally negative for NM myosin, indicating a maturation toward adult SM cells. Concomitant to this phenotypic change, the response to KCl increased, and a bethanechol-stimulated contractile response developed. At no time period did the serosal layer react with anti-synaptophysin or anti-neurofilament proteins nor did the strips respond to field stimulation (via release of neurotransmitters), showing that SM cell differentiation and development of contractile responses during serosal thickening are independent of innervation.

Animals↗
PubMed1980

Evidence of similar organization of the chromosomes carrying the major histocompatibility complex in man and other primates.

The chromosome localization and gene synteny of the major histocompatibility complex (MHC) of the great apes and rhesus monkey were investigated using somatic cell hybrids. The presence of the MHC antigens was determined either with a microadsorption technique employing primate alloantisera, or with a radioimmune assay. The enzymes phosphoglucomutase 3 (PGM3), glyoxalase 1 (GLO1), mitochondrial superoxide dismutase (SOD2), and soluble maleic enzyme (ME1) were assayed in those hybrids where electrophoretic separations could be achieved. A chromosome homologous to the human No. 6 was found in the chimpanzee, gorilla, orangutan and rhesus monkey, and its genomic organization is similar to that of man.

Animals↗
PubMed1983

Complex chromosome homologies between the rhesus monkey (Macaca mulatta) and man.

The chromosome localization and gene synteny of soluble malate dehydrogenase (MDH1), soluble isocitrate dehydrogenase (IDH1), mitochondrial superoxide dismutase (SOD2), phosphoglucomutase-3 (PGM3), mitochondrial malate dehydrogenase (MDH2), beta-glucuronidase (GUSB), nucleoside phosphorylase (NP), pyruvate kinase M2 (PKM2), hexosaminidase A (HEXA), inosine triphosphatase (ITPA), and N-acetyl-alpha-D-galactosaminidase (NAGA) were determined in the rhesus monkey using somatic cell hybrids. Comparison with the human and Pongidae syntenic groups shows that chromosome banding homologies do not always correlate with gene mapping data.

Animals↗
PubMed1992

Linkage of two enzyme loci in the fish genus Poecilia (Teleostei:Poeciliidae).

The linkage of loci coding for glucose-6-phosphate dehydrogenase (G6PD) and phosphogluconate dehydrogenase (PGD) is described in fish of the genus Poecilia (Teleostei:Poeciliidae) and designated Poecilia linkage group I. These two loci were shown to assort independently from six other informative markers (peptidase S, malate dehydrogenase 2 [soluble], mannose phosphate isomerase, parvalbumin 2, phosphoglucomutase, and glyceraldehyde-3-phosphate dehydrogenase 2) within the limits of the data obtained. Data for the linkage analyses were generated by scoring starch-gel electrophoretic phenotypes of the eight loci in reciprocal backcross hybrids obtained from matings between Poecilia perugiae and P. vittata. The linkage chi 2 for G6PD-PGD locus pairs was significant (P less than 0.001) in all reciprocal backcross hybrid broods (22.7% recombinants in the combined data), indicating linkage in both parental species. The linkage of G6PD and PGD in gene maps of the poeciliid genera Xiphophorus and Poeciliopsis documents homology of this linkage within the family. Linkages in salmonid and centrarchid fishes suggest conservation of this linkage group in most or all teleosts. The six additional indpendently assorting loci have been assigned to independent linkage groups in Xiphophorus; thus, no example of poeciliid linkage group divergence has yet been identified.

Animals↗
PubMed1997

Chromosome location of sixteen genes in the common shrew, Sorex araneus L. (Mammalia, Insectivora).

This report extends the genetic map of the common shrew (Sorex araneus) by use of a clone panel of shrew-Chinese hamster and shrew-mouse hybrid cells (Pack et al., 1995; Matyakhina et al., 1996). This set of hybrid clones made it possible to assign the shrew genes for isocitrate dehydrogenase 2 (IDH2), inorganic pyrophosphatase (PP), glutamicpyruvate transaminase (GPT), adenosine kinase (ADK), glucuronidase 2 (GUSB) and acid phosphatase 2 (ACP2) to chromosome ik; the genes for adenylate kinases 1 and 3 (AK1 and AK3) to chromosome af; the genes for glutamate-oxaloacetate transaminase 2 (GOT2), peptidase D (PEPD) and growth hormone (GH) to chromosome hn; the gene for phosphoglucomutase 2 (PGM2) to chromosome go, the gene for enolase 1 (ENO1) to chromosome ji, the gene for ornithine carbamoyl-transferase (OTC) to chromosome de, the gene for aminoacylase 1 (ACY1) to arm m (chromosome mp), the gene for glutamate-oxaloacetate transaminase 1 (GOT1) to arm q (chromosome qr). Thus, the genetic map of the common shrew now contains 33 genes and it is possible to compare the syntenic associations with other species.

Animals↗
PubMed1977

The use of the FST statistic of Wright for estimating the effects of genetic drift, selection and migration populations, with special reference to Ireland.

The Fst of Wright has been used to examine the available blood group, serum protein and enzyme data for the world, NW Europe and the counties of Ireland. These include the ABO, secretor, Lewis, MNSs, Rh, Kell, Duffy, Lutheran, Kidd, P, Diego, haptoglobin, Gc, Lp, Ag, adenosine deaminase, adenylate kinase, acid phosphatase, 6-phosphogluconate, phosphoglucomutase and transferrin systems. The highest value was found for the Fy gene. Much lower values than those calculated for world data were found for NW Europe and Ireland with the exception of the Lpa antigen which had high values in Ireland. sigma2p was used to estimate rates of genetic drift in an Irish population and it was estimated that a migration rate of 4% would counter genetic drift in Ireland.

Blood Group Antigens↗
PubMed1979

The inherited blood factors of some Northern Nigerians.

Results are presented on 147 individuals from northern Nigeria who were tested for the red cell antigens A, A1, B, H, M, N, S, s, He, P1, C, D, Du, E, c, e, Ce, v, Lua, Jka (some for Jkb), Lua, K, Jsa (some for Jsb), Kpa, Rd, Fya and Fyb, and for variants of the serum proteins haptoglobin and transferrin and of the red cell enzymes acid phosphatase, phosphoglucomutase, glucose-6-phosphate dehydrogenase, adenylate kinase, adenosine deaminase, phosphohexose isomerase and lactate dehydrogenase. The results found are of interest as they are among the very few published for this area of Nigeria, but they show little that is unexpected for people living in this region.

Blood Group Antigens↗
PubMed1982

Distribution of serum proteins, red cell enzymes and haemoglobins in vitiligo.

125 Sudanese patients suffering from vitiligo were investigated for the distribution of serum proteins (haptoglobins and transferrins), red cell enzymes (acid phosphatase, 6-phosphogluconate dehydrogenase, phosphoglucomutase and glucose-6-phosphate dehydrogenase) and hemoglobins. The results were compared with the published healthy population series investigated for the same genetic markers. There was no significant association with any of the marker systems in vitiligo except glucose-6-phosphate dehydrogenase. An excess deficiency of this enzyme was observed in vitiligo patients compared to the control series.

Blood Proteins↗
PubMed1983

Blood groups, haptoglobins and red cell isoenzymes of the Jat Sikhs of Ludhiana District, Panjab, India.

Blood samples from 156 Jat Sikhs were examined for the ABO, MNS, Rhesus, Duffy and Kell blood groups. Serum and lysate samples were prepared and electrophoretically tested for haptoglobin, esterase D, adenylate kinase, phosphoglucomutase and the acid phosphatase systems. Comparisons were made with other populations of the Indian sub-continent and of contiguous regions.

Blood Group Antigens↗
PubMed1983

Genetic studies in Poland.

Blood specimens were studied from 213 persons aged 3-51 years, in Olkusz, a town between Krakow and Katowice in south Poland. The data show general similarity in blood groups and red cell enzymes to those from other central European populations, but in several systems there are suggestions that Polish frequencies are somewhat peripheral, for example phosphoglucomutase, acid phosphatase, esterase D, while in others where clines exist the results fit well into those expected from the geographical position of the sample.

Adolescent↗
PubMed1984

Genetic polymorphisms in the Kuwaiti Arabs.

Blood samples were collected from 162 Kuwaiti Arabs. These samples were typed for the ABO, MNSs, Rh, Kell and Duffy blood group systems, serum protein haptoglobins, the red cell isoenzymes acid phosphatase, phosphoglucomutase (locus 1), adenylate kinase, 6-phosphogluconate dehydrogenase, and the lactate and malate dehydrogenase variants. Comparisons were made with serological findings for other Arab populations in the Arabian peninsula.

Blood Group Antigens↗