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Biomedical subjects

S Ray

Publications and source records attributed to S Ray.

At least 37 records · Page 2Linked to original sources

Developmental expression of the embryonic chicken brain DNA polymerase alpha and its binding with monoclonal antibodies against human KB cell DNA polymerase alpha.

Changes in DNA polymerase alpha activity accompanying tissue development have been well established in several systems. In most cases, DNA polymerase alpha activity decreases with development. Here, we report observed changes in DNA polymerase alpha activity throughout embryonic chicken brain (ECB) development. The level of DNA polymerase alpha activity was found to gradually decrease by 60% (2.3 to 0.8 nmol of [3H]dCMP incorporated/mg protein/h) between 9- and 19-day-old ECB. An enzyme-linked immunosorbent assay of DNA polymerase alpha utilizing monoclonal antibody SJK 237-71 (human KB cell DNA pol-alpha binder) also demonstrated a gradual decrease (up to 60%) of antigen over this same range of development. Analysis of DNA polymerase alpha from 11- and 19-day-old ECB by a 10 to 30% glycerol density gradient revealed a high molecular weight peak sedimenting near catalase (11.3 S) with activity at the 11th day being approximately 3-fold greater than activity at the 19th day. A Western immunoblot analysis utilizing monoclonal antibody SJK 237-71 (against human KB cell DNA polymerase alpha) showed a decrease in DNA polymerase alpha from 186 kilodaltons in 9- and 11-day ECB cell-free extracts to 120 kilodaltons in extracts from 13- to 19-day ECB. The conversion of DNA polymerase alpha from a higher to a lower molecular weight form may be a regulatory mechanism in eukaryotic DNA replication.

Animals

A PMA degrading constitutive organomercurial lyase in a broad-spectrum mercury resistant Bacillus pasteurii strain DR2.

A broad-spectrum Hg-resistant strain of B. pasteurii DR2 utilized phenylmercuric acetate (PMA) as sole source of carbon. This bacterial strain contained a constitutive organomercurial lyase which specifically degraded PMA but not other organo-mercurials. This PMA-lyase activity was also stimulated to different extents when this bacterial strain was grown in presence of different organic compounds as sole source of carbon.

Bacillus

Role of arginine 67 in the stabilization of chymotrypsin inhibitor 2: examination of amide proton exchange rates and denaturation thermodynamics of an engineered protein.

We have examined the contribution to protein stability of an interaction involving a charged hydrogen bond from an arginyl side chain (Arg67) in the serine proteinase inhibitor chymotrypsin inhibitor 2 (CI-2), by replacing this side chain with an alanyl residue by protein engineering. Using nuclear magnetic resonance spectroscopy (NMR), we have examined the effect of this mutation on the hydrogen-deuterium exchange rates of several backbone amide protons in the native and engineered proteins at 50 degrees C. These exchange rates provide a localized probe at multiple discrete sites throughout the protein and from comparison of native and mutant exchange rates allow calculation of the difference in free energy of exchange (delta delta Gex) resulting from the mutation. The results show that for the majority of amides observed this mutation results in delta delta Gex of ca. 1.7 kcal mol-1 over the whole CI-2 molecule. However, for two relatively exposed amide protons the exchange rates are found to be far less perturbed, implying that local unfolding mechanisms predominate for these protons. Direct measurement of the stability of both proteins to denaturation by guanidinum hydrochloride shows that the interaction contributes 1.4 kcal mol-1 to the stability of the molecule. This value is comparable to those obtained from the NMR exchange measurements and indicates that the exchange processes reflect the differences in stability between the native and mutant proteins.(ABSTRACT TRUNCATED AT 250 WORDS)

Arginine

Monomer sequence determination of carbohydrates using fast-atom bombardment mass spectrometry of periodate-oxidized acetate ester derivatives.

A derivatization method, adapted from that of Angel et al. (ref. 10), for sequencing sugar residues in partially degraded poly- and oligo-saccharides using positive-ion f.a.b.-m.s. is described. Derivative selection provides sequence information by directing fragmentation exclusively to both sides of glycosidic oxygen atoms and, in the case of opened rings, between glycosidic carbon and ring oxygen atoms. Polysaccharides or oligosaccharides are subjected to sequential periodate oxidation, borodeuteride reduction, and acetylation. The derivatized polysaccharides are then subjected to partial degradation, acetylation, and high-performance liquid chromatography (h.p.l.c.) purification. F.a.b.-m.s. data obtained on model compounds, using 3-nitrobenzyl alcohol as matrix for f.a.b.-m.s., demonstrated direction of fragmentation to both sides of the glycosidic oxygen atom in unoxidized residues, and to both sides of the acetal oxygen atoms in oxidized residues. Oligosaccharide linkage and sequence may thus be determined by observing fragmentation from both the reducing and non-reducing ends of the molecule. Two Salmonella lipopolysaccharides, derivatized by this procedure, were partially hydrolyzed and then acetylated. Analysis of the h.p.l.c.-purified oligosaccharide derivatives by f.a.b.-m.s. demonstrated the applicability of the technique for sequencing nmol quantities of branched structures.

Benzyl Alcohols

A pilot study of quinidine and epirubicin in the treatment of advanced breast cancer.

Thirty-one patients were entered into a pilot study combining oral quinidine with epirubicin 100 mg m-2 as first line chemotherapy in advanced breast cancer. Three patients were treated with quinidine 1 g b.d., and developed symptoms of toxicity. Of eight subsequent patients treated with quinidine 500 mg b.d., two experienced tiredness and nausea and one severe oral toxicity with epirubicin. The remaining 20 patients received quinidine 250 mg b.d.; one developed cinchonism and one malaise, the remainder showing no excess toxicity compared with epirubicin alone. The median nadir WBC was similar with or without quinidine (2.3 vs 1.6 x 10(9) l-1) as was median nadir platelet count (175 vs 157 x 10(9) l-1). There was no evidence of significant cardiac toxicity. The median plasma quinidine level achieved was 5.6 mumol l-1 (range 2.1-22.1), which is within the range of concentrations which is effective in vitro at reversing experimental anthracycline resistance. A randomised controlled study is proposed to assess the impact of this potential modulation on the efficacy of epirubicin in advanced breast cancer.

Adult

Scoliosis in trisomy 18.

Patients with trisomy 18 typically present with multiple congenital anomalies and most die within the first year. However, long-term survivors are not uncommon. Seventeen patients with trisomy 18 were evaluated to study the development of scoliosis associated with this disorder. There were 13 females and 4 males with ages ranging from birth to 22 years. Twelve patients died by age 2. None developed scoliosis or had vertebral anomalies. The five patients who survived beyond age 2 developed scoliosis. Curve progression was demonstrated in the patients who returned for follow-up. Bracing was not well tolerated in two patients with curves of 48 degrees and 58 degrees. Both had poor motor control and sitting ability. One patient with a 30 degree curve was successfully managed by bracing. Another patient with a severe scoliosis was successfully fused with anterior and posterior instrumentation. Patients with trisomy 18 should be carefully evaluated for scoliosis. Scoliosis in the older child, surviving beyond age 2, may be progressive and difficult to manage.

Abnormalities, Multiple

Novel exercise protocol suitable for use on a treadmill or a bicycle ergometer.

Many exercise protocols are in use in clinical cardiology, but no single test is applicable to the wide range of patients' exercise capacity. A new protocol was devised that starts at a low workload and increases by 15% of the previous workload every minute. This is the first protocol to be based on exponential rather than linear increments in workload. The new protocol (standardised exponential exercise protocol, STEEP) is suitable for use on either a treadmill or a bicycle ergometer. This protocol was compared with standard protocols in 30 healthy male volunteers, each of whom performed four exercise tests: the STEEP treadmill and bicycle protocols, a modified Bruce treadmill protocol, and a 20 W/min bicycle protocol. During the two STEEP tests the subjects' oxygen consumption rose gradually and exponentially and there was close agreement between the bicycle and the treadmill protocols. A higher proportion of subjects completed the treadmill than the bicycle protocol. Submaximal heart rates were slightly higher during the bicycle test. The STEEP protocol took less time than the modified Bruce treadmill protocol, which tended to produce plateaux in oxygen consumption during the early stages. The 20 W/min bicycle protocol does not take account of subjects' body weight and consequently produced large intersubject variability in oxygen consumption. The STEEP protocol can be used on either a treadmill or a bicycle ergometer and it should be suitable for a wide range of patients.

Adult

Studies on synergism between penicillins and ambodryl (bromodiphenhydramine HCl), an antihistamine with antimicrobial property.

With respect to 31 different selected test bacteria, all sensitive to benzyl penicillin (Pc), ampicillin (Ap), methicillin (Me), ceporan (Ce), cloxacillin (Cx), streptomycin (Sm), kanamycin (Km), gentamicin (Gm), chloramphenicol (Cm), tetracycline (Tc), polymyxin (Pm) as well as ambodryl [Am; an antihistamine (bromodiphenhydramine HCl) with distinct antimicrobial properties], it was found that Am in combination either with Pc, Ap, Ce or Me consistently showed enhancement of antimicrobial effects resulting from synergism. A combination of Am with either Sm, Km, Gm or Tc, on the other hand, showed only additive effects. An interaction of the activities of Am with Pm also resulted in indifference effects. Determination of the area of inhibition zone, calculated from its diameter for the degree of synergism in case of Am and Pc, showed these synergistic effects to be significant (P less than 0.05) in comparison with their individual effects: this was corroborated by the determination of the fractional inhibitory concentration (FIC) index which was found to be less than 0.5. The synergism of Am-Pc combination was confirmed by in vivo studies by challenging mice with a virulent strain of S. typhimurium and looking for protection.

Bacteria

Acid excretion and serum electrolyte patterns in patients with advanced chronic renal failure.

Hyperchloremic metabolic acidosis can be seen in advanced chronic renal failure (CRF). To study this further, we measured acid excretion under controlled conditions in 19 patients with severe but stable CRF. Twelve patients had a hyperchloremic metabolite acidosis, 3 had an elevated anion gap acidosis ('delta acidosis'), while the 4 remaining patients had a slightly decreased tCO2 with a normal anion gap and serum chloride level ('mild acidosis'). Ammonium excretion was markedly reduced in CRF patients (11 +/- 1 vs. 32 +/- 2 mEq/day in normal subjects, p less than 0.005), and likewise urinary titratable acid was diminished (18 +/- 2 mEq/day in patients, vs. 28 +/- 3 mEq/day in normal subjects; p less than 0.005). When expressed per 100 ml GFR, ammonium and titratable acid excretion were markedly increased in all groups of patients compared to normal subjects. The Tm for bicarbonate was 22.5 +/- 3.7 mEq/l for all CRF patients and 24 +/- 1 mEq/l for the patients with a hyperchloremic metabolic acidosis. We conclude that the hyperchloremic metabolic acidosis in advanced renal failure is due to diminished excretion of ammonium and titratable acid and is not due to an increased bicarbonate leak. As renal failure advances and renal mass declines, the remaining functioning nephrons hypersecrete acid.

Acidosis

Pattern of acute myocardial infarction in a district hospital in Brunei Darussalam--a pilot study.

The objective of the authors was to study the pattern of presentation, risk factors, and natural course of acute myocardial infarction in the general population of Belait District in Brunei Darussalam. A prospective study was done of 100 consecutive cases of acute myocardial infarction admitted to the coronary care beds of a District General Hospital. The patients were followed up to 12 weeks after admission to hospital. There were three times more males than females (75 males, 25 females). Nine cases out of 75 males were below the age of 40 years, 3 being below 30 years. The mean age of the male denominator was 57.4 years while that of the female counterpart was 67.48 years showing a mean difference of 10.08 years. There were significant association with hypertension (31%), smoking (30%) and diabetes mellitus (27%). The majority of the patients had prodromal symptoms, the most common presenting symptoms were chest pain (63%) and shortness of breath (27%). Only 4% of the patients had silent infarction. Acute myocardial infarction is common in Brunei forming 2.6% of all the patients admitted to the medical wards and the relative rate is 3 times higher in males than in females. There is significant association of IHD with 3 main risk factors namely hypertension, smoking and diabetes mellitus.

Adult

Physiological changes in rats after exposure to low levels of microwaves.

The effects of exposure to sublethal levels of microwaves were studied. Young albino rats of both sexes were exposed for 60 days to 7.5-GHz microwaves (1.0-KHz square wave modulation, average power 0.6 mW/cm2) for 3 h daily. During and after microwave exposure several physiological parameters were measured in both control and exposed animals. It was found that the animals exposed to microwaves tended to eat and drink less and thus showed a smaller gain in body weight. Some of the hematological parameters and organ weights were also significantly different. It is proposed that a nonspecific stress response due to microwave exposure and mediated through the central nervous system is responsible for the observed physiological changes.

Animals

Prevalence of hepatitis-B surface antigen in the pregnant women of Brunei Darussalam.

In view of the high incidence of mother-to-child transmission of hepatitis B virus (HBV), routine screening of pregnant at antenatal clinic and vaccination of the newborn has become a common practice throughout the world. The prevalence of HBsAg in pregnant women of Brunei Darussalam was assessed. Of 1,267 pregnant women who attended the antenatal clinic of Suri Seri Begawan Hospital, Kuala Belait, the overall prevalence of HBsAg was 3.2% with significant differences in various ethnic groups (p less than 0.05). Malays, who form the major ethnic group had the lowest rate of 1.9%.

Adolescent

Methylglyoxal-catabolizing enzymes of Leishmania donovani promastigotes.

Methylglyoxal is a toxic metabolite with growth inhibitory properties against Leishmania donovani promastigotes. We have shown in the present study that both log and stationary phase promastigotes of L. donovani can catabolize methylglyoxal to D-lactate as the major end product. The specific activity of methylglyoxal reductase was found to be the highest of all the catabolic enzymes. In contrast, the anabolic pathway for methylglyoxal could not be detected. Moreover, when control promastigotes or promastigotes in which the glycolytic pathway was inhibited were incubated with glucose, glycerol or dihydroxyacetone phosphate as energy source, neither methylglyoxal nor D-lactate could be detected.

Alcohol Oxidoreductases

Accuracy of inpatient oxygen administration.

The accuracy of supplemental oxygen delivery was assessed in two medical units. Only 90 of 206 patients (44%) were receiving oxygen as prescribed. The ward oxygen rotameters were inaccurate at normal clinical flow rates, the range of flows being delivered as a percentage of the indicated flow varying from 15% at 8 l/min to 40% at 1 and 2 l/min.

Equipment Design