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Biomedical subjects

M Mukherjea

Publications and source records attributed to M Mukherjea.

At least 19 recordsLinked to original sources

Enzymatic tumour markers in ovarian cancer: a multiparametric study.

The serum concentration of some enzymes, namely placental alkaline phosphatase, lactate dehydrogenase, 5' nucleotidase and amylase, was determined by commercially available kit in 50 ovarian cancer patients and 31 patients with benign gynaecological disease before initiation of any treatment. The values were compared with those of 30 healthy women. Multivariate analysis showed a statistically significant difference between healthy women and ovarian cancer patients. These results indicate that by using discriminating function of the above four enzyme variables it is possible to screen ovarian cancer in outpatient obstetric and gynaecological clinics (sensitivity 96%, specificity 83.3%, relative risk 11.7). Hence this system can serve as a suitable marker for ovarian cancer.

5'-Nucleotidase

Role of rusticyanin in the electron transport process in Thiobacillus ferrooxidans.

Effect of diethyl dithiocarbamate (DEDC), an antimicrobial agent, on growth of Thiobacillus ferrooxidans, possibly by inhibiting rusticyanin present in the periplasmic space of the microorganism, has been studied to gain more insight into the electron transport chain in the bioleaching process. DEDC is found to form a stable complex with rusticyanin in solution and also in polyacrylamide gel. The spectrum of the complex is identical to that of Cu-DEDC complex, suggesting binding of DEDC with copper moiety of rusticyanin and resulting in inhibition of growth. In vitro reduction of purified rusticyanin by Fe(II) in absence of acid-stable cytochrome c is very slow, indicating the importance of cytochrome c in electron transport. Thus, in the iron oxidation process, acid-stable cytochrome c is the primary acceptor of electron, transferring the electron to rusticyanin at pH 2.0, which, in turn, affects electron transfer to iron-cytochrome c reductase around pH 5.5.

Azurin

Relationship between fatty acid synthesis, transport and total lipid content during human fetal lung development.

Levels of fatty acid binding proteins (FABPs), lipids as well as activities of fatty acid synthesizing enzymes such as fatty acid synthase and ATP-citrate lyase increase with gestation showing maximum at term in human fetal lung. However, the activity of ATP-citrate lyase showed the same trend up to 30 weeks of gestation before declining slightly at term. These results indicate the importance of supply and/or synthesis of fatty acids when lung surfactant synthesis begins; thereby showing a correlation between the FABPs, lipid pattern and the activities of fatty acid synthesizing enzymes during prenatal lung development.

ATP Citrate (pro-S)-Lyase

Human fetal liver fatty acid binding proteins. Role on glucose-6-phosphate dehydrogenase activity.

Fatty acid binding proteins (FABPs) may play an important role in the transport and metabolism of fatty acids during human embryogenesis. Three fractions of FABP, namely, DE-I, DE-II and DE-III, having Mr 14,200 Da each and pI values 7.8, 6.9 and 5.4, respectively, have been detected in human fetal liver. These proteins were purified by heat and butanol precipitation of fetal liver supernatant as well as by gel filtration and ion-exchange chromatography. Fetal liver FABPs are immunochemically identical to each other. Concentrations of DE-I, DE-II and DE-III increase gradually from early gestation to term. DE-I is almost lipid-free, DE-II binds long-chain fatty acids nonspecifically and DE-III transports mainly arachidonic acid. DE-II and DE-III protect glucose-6-phosphate dehydrogenase, which furnishes NADPH for fatty acid synthesis, from the feed-back inhibition exerted by added palmitoyl-CoA and oleate. In the absence of exogenous inhibitors, this enzyme is stimulated by FABPs. DE-I has no effect on such inhibition. Thus, FABPs play a regulatory role in critical aspects of cellular physiology during human embryogenesis.

Carrier Proteins

Purification and characterization of fatty acid-binding proteins from human fetal lung.

Fatty acid-binding protein (FABP) was isolated, purified, and characterized from developing human fetal lung cytosol by gel filtration and ion-exchange chromatography. FABP exists in three immunochemically identical forms, DE-I, DE-II, and DE-III, having Mr 15,200 +/- 200 each and isoelectric pH 7.8, 6.9, and 5.4, respectively. DE-I is almost lipid-free, DE-II binds mainly long-chain unsaturated fatty acids, and DE-III is an arachidonic acid carrier. One mole of DE-II and DE-III each binds 1 mol of fatty acids noncovalently. Concentrations of all these FABPs increase gradually from early gestation to term. Defatted lung FABP reverses the inhibitory effect of palmitoyl coenzyme A (CoA) (PAL-CoA) on lung glucose-6-phosphate dehydrogenase (G6PD), a key enzyme of the hexose monophosphate (HMP) shunt pathway. This protein when added alone activates the enzyme, suggesting that the original submaximal activity is probably due to the presence of endogenous long-chain fatty acyl CoA esters in the cytosols. As FABP is present in relatively high concentration in most mammalian cells, the potent inhibitory effects of long-chain acyl CoA esters on the HMP shunt pathway in vitro are not seen in intact cells.

Carrier Proteins

Purification and characterization of fatty acid-binding protein from human placenta.

Purification of a cytosolic fatty acid-binding protein (FABP) from developing human placenta has been achieved, and its role in modulating the inhibition of human placental glucose-6-phosphate dehydrogenase (G6PD) by palmitoyl-CoA (PAL-CoA) has been studied. FABP was resolved into three peaks, viz. DE-I, DE-II and DE-III, by DEAE cellulose chromatography. DE-I was almost lipid-free. Presence of endogenous fatty acids in DE-II and DE-III was detected by thin layer chromatography (TLC). Fatty acids were the only detectable lipid component in these fractions. Gas liquid chromatography (GLC) analysis revealed that DE-II binds long chain saturated and unsaturated fatty acids nonspecifically, whereas DE-III is mainly an arachidonic acid carrier. Each of these fractions, viz. DE-I, DE-II and DE-III, has a molecular weight of 14,200 Daltons. Ouchterlony double immunodiffusion studies have confirmed the immunochemical identity of these three fractions of placental FABP. Separation in ion exchanger may be due to their different isoelectric points and varied types of binding affinities. Human placental G6PD was inhibited 50% by 0.03 mM PAL-CoA. The DE-II fraction of FABP enhanced the activity of G6PD in the absence of added PAL-CoA and protected against PAL-CoA inhibition of the enzyme. Such a modulating effect of FABP in this inhibition is attributable to binding of long chain acyl-CoA rather than to a direct effect of FABP on the enzyme itself.

Carrier Proteins

Histological changes in the ovary and uterus of rat after injectable contraceptive therapy.

Histological changes were studied in the ovary and uterus of rats receiving different doses of Depo-Provera (DMPA) and Noristerat (NET-EN) for varying duration. The effect of DMPA on ovarian and uterine tissues was strongly progestational. The whole morphological alteration in the ovary after DMPA therapy appeared to be the atresia of the follicular apparatus with degeneration of the growing follicles. Uterine histology reflected that endometrial tissues gradually became inactive and with prolonged treatment at high doses, atrophy of the endometrium was noted. In NET-EN-treated rats, absence of mature follicles and recent corpora lutea reflected the blockade of ovulation. There was no extreme atrophy of the ovary or endometrium as found with DMPA treatment. With higher doses of NET-EN, endometrial growth was arrested.

Animals

Alterations in protein, sialic acid and some enzymes in cervical mucus of female rats during NET-EN treatment.

The effect of norethisterone enanthate (NET-EN) on cervical mucus protein, sialic acid and some enzymes (e.g. peroxidase, alkaline phosphatase and alpha-amylase) were studied in adult female rats. One mg NET-EN every 12 days was found to be an effective contraceptive dose of this drug in this species, acting primarily through the cervical mucus. NET-EN produced a highly significant increase in protein content and peroxidase and alkaline phosphatase activities. However, sialic acid content and amylase activity did not exhibit any definite pattern after NET-EN therapy. The increased protein content together with persistent elevated levels of peroxidase and alkaline phosphatase corroborates the hypothesis that NET-EN creates a progestogenic phase which prevents sperm penetration and thus conception.

Alkaline Phosphatase

Characterization of microsomal ATPases from developing human placenta.

Activities and some properties of microsomal ATPases have been studied in developing human placenta. The enzyme activities (Na+ + K+ + Mg2+, Mg2+, and Ca2+ dependent) in the placenta increase steadily with gestational age until the 18th to 21st week, and decrease in the second half of pregnancy. Mg2+-dependent and Na+ + K+ + Mg2+-dependent ATPases possess nearly the same Km (apparent) for ATP, while the Ca2+-dependent enzyme shows a different one. Mg2+-dependent ATPase shows higher substrate affinity than Ca2+-dependent ATPase, although the Vmax of the Mg2+-dependent enzyme is lower than that of the latter. However, for each enzyme, the Km remains almost constant and Vmax varies during ontogenic development. Vmax of the enzymes decline at term. The enzymes are heat-labile, unaffected by amino acids, namely, L-phenylalanine, L-leucine, and L-tryptophan, and deoxycholate inhibits the enzyme activities by about 50%.

Adenosine Triphosphatases

Effect of norethisterone enanthate on the blood count and endometrial histology of Indian women.

Endometrial histology and the haematological indices were studied in women receiving 200 mg of norethisterone enanthate (NET-EN) at 60-day intervals for 24 months as a method of contraception. Significant elevations occurred in the haemoglobin concentration and red blood cell counts during one year of therapy, while eosinophil counts showed a transient increase at 4 months of trial. The changes in the haematological indices seem to be due to stimulation of the erythropoietic system by NET-EN and may also be attributable to diminished blood loss due to amenorrhoea during the course of therapy. The endometrial histology revealed proliferative glands at 2 months which turned quiescent at 6-12 months of NET-EN therapy. Long-term use of 18 months showed atrophic changes with few glands, narrow lumina and stromal oedema.

Blood Cells

A comparative study of human placental and fetal liver catalase during development.

The activity and a few properties of catalase have been compared in the developing human placenta and fetal liver. The presence of the enzyme in both the tissues is discernible as early as in the 6th wk of gestation and the activity increases gradually with the advancement of pregnancy. Maximum enzyme activity in both placenta and fetal liver is found to be associated with the soluble supernatant fraction obtained by centrifuging the tissue homogenates at 105000 X g. Kinetic studies reveal the enzymatic decomposition of H2O2 to follow first-order kinetics at lower substrate concentrations, and then to deviate from the original linearity, demonstrating mixed-order kinetics. Thermostability of placental catalase increases with prenatal development, while the enzyme from fetal liver remains moderately heat-stable throughout the gestation. Treatment of the homogenates with Triton X-100 is found to be most effective in increasing catalase activity in each of these tissues.

Catalase

Studies on some enzymes and sialic acid during progestational contraceptive therapy.

Injectable progestogen, norethisterone enanthate (NET-EN, 200 mg/ml at 60-day intervals), was administered to one-hundred-fifty women for two years as a method of contraception. Blood levels of acid phosphatase (ACP), alkaline phosphatase (AP), glutamate pyruvate transaminase (GPT), glutamate oxaloacetate transaminase (GOT), acetylcholinesterase (AChE) and sialic acid were determined in all the subjects to ascertain whether NET-EN therapy causes any adverse metabolic effect or damage to the functional status of the liver. NET-EN contraception did not alter the liver function enzymes, but there is a significant increase (P less than 0.001) in AChE activity after two years. Serum sialic acid level showed a transient increase up to one year, which however returned to control level later. The mechanism responsible for these changes and whether the rise in sialic acid and AChE activity are related to any pathological condition remain unclear at this stage.

Acetylcholinesterase

Action of polyamines on ribonucleic acid and protein synthesis during ontogeny of human fetal liver.

A higher rate of RNA synthesis in human fetal liver cells was found during the early age of gestation, after a drop, it gradually increases to a maximum value between 18 and 22 weeks, followed by a sharp decrease at later period of gestation. Total polyamine content of fetal liver tissue shows a similar trend. However, when liver cell suspension was incubated with exogenous spermine, spermidine and putrescine there is a dose-dependent inhibition in RNA synthesis. Protein synthesis in human fetal liver cells was stimulated with lower doses of spermine, spermidine and putrescine and inhibited at higher doses. The optimum dose for stimulation and the degree of stimulation was, however, not the same for fetuses of different gestational ages.

Fetus