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Biomedical subjects

B Das

Publications and source records attributed to B Das.

At least 37 records · Page 2Linked to original sources
PubMed2001

Uropathogenic Escherichia coli causing urinary tract infections.

BACKGROUND & OBJECTIVES: The information on the characteristics of Escherichia coli causing urinary tract infections is limited. We have characterised the urovirulence factors of Esch. coli isolated from symptomatic patients of urinary tract infections (UTI) in order to determine their pathogenic potential and the antibiotic sensitivity profile. METHODS: Semi-quantitative urine culture was done on 370 symptomatic patients suffering from urinary tract infections. Phenotypic characterization of the urovirulence factors of Esch. coli was undertaken and the antibiotic susceptibility was determined. RESULTS: Esch. coli was responsible for 45.5 per cent of infections. Resistance to amoxycillin, cotrimoxazole, nalidixic acid, norfloxacin and ciprofloxacin among Esch. coli isolates ranged from 70-95 per cent. Serotype O101 was found to be the commonest serotype (7/26). The virulence factors associated with uropathogenic Esch. coli were haemolysin production (5/30), presence of mannose resistant P-fimbriae (5/30), presence of mannose sensitive type 1 fimbriae (6/30) and the presence of mannose resistant F-fimbriae (2/30). Siderophores production was seen in all the isolates causing UTI. INTERPRETATION & CONCLUSION: Esch. coli was found to be the commonest cause of UTI in our study population. Antibiotic resistance was high among the strains circulating which emphasises the need for judicious use of antibiotics. Certain virulence factors like haemolysin production and presence of fimbriae in the Esch. coli may be associated with the urovirulence.

Anti-Bacterial Agents↗
PubMed2000

Multicopy molecular dynamics simulations suggest how to reconcile crystallographic and product formation data for camphor enantiomers bound to cytochrome P-450cam.

Multiple ligand binding modes are possible in many enzyme active sites; their presence in cytochrome P450cam (P450cam) is evident from crystallographic studies of the binding of thiocamphor and phenylimidazoles. Here, we use multicopy molecular dynamics simulations to compare the binding modes of (1R)- and (1S)-camphor in the active site of P450cam. Simulations with (1R)-camphor, the natural substrate, serve to calibrate our protocol: 19 out of 20 copies of (1R)-camphor converged to coordinates very close to those observed for (1R)-camphor in its crystallographic complex with P450cam during the simulations. Simulations with the (1S)-camphor enantiomer showed greater mobility of the substrate, consistent with spectroscopic data, and resulted in 3 major binding modes. One of these is similar to the major conformation (of the two conformations assigned) in a recently determined crystal structure, but this conformation is not correctly oriented for regiospecific hydroxylation at C-5. The simulations, however, provide evidence for reorientation of (1S)-camphor upon formation of the reactive Fe-O intermediate to an orientation suitable for hydroxylation. The simulations thus permit rationalisation of the apparent inconsistency between the crystal structure and the reaction products.

Binding Sites↗
PubMed2000

Cytochrome c methyltransferase, Ctm1p, of yeast.

Cytochromes c from plants and fungi, but not higher animals, contain methylated lysine residues at specific positions, including for example, the trimethylated lysine at position 72 in iso-1-cytochrome c of the yeast Saccharomyces cerevisiae. Testing of 6,144 strains of S. cerevisiae, each overproducing a different open reading frame fused to glutathione S-transferase, previously revealed that YHR109w was associated with an activity that methylated horse cytochrome c. We show here that this open reading frame, denoted Ctm1p, is specifically responsible for trimethylating lysine 72 of iso-1-cytochrome c. Unmethylated forms of cytochrome c but not other proteins or nucleic acids are methylated in vitro by Ctm1p produced in S. cerevisiae or Escherichia coli. Iso-1-cytochrome c purified from a ctm1-Delta strain is not trimethylated in vivo, whereas the K72R mutant form, or the trimethylated Lys-72 form of iso-1-cytochrome c, are not significantly methylated by Ctm1p in vitro. Like apocytochrome c, but in contrast to holocytochrome c, Ctm lp is located in the cytosol, consistent with the view that the natural substrate is apocytochrome c. The ctm1-Delta strain lacking the methyltransferase did not exhibit any growth defect on a variety of media and growth conditions, and the unmethylated iso-1-cytochrome c was produced at the normal level and exhibited the normal activity in vivo. Ctm1p and cytochrome c were coordinately regulated during anaerobic to aerobic transition, a finding consistent with the view that this methyltransferase evolved to act on cytochrome c.

Amino Acid Sequence↗
PubMed2000

Control of intramolecular interactions between the pleckstrin homology and Dbl homology domains of Vav and Sos1 regulates Rac binding.

Vav and Sos1 are Dbl family guanine nucleotide exchange factors, which activate Rho family GTPases in response to phosphatidylinositol 3-kinase products. A pleckstrin homology domain adjacent to the catalytic Dbl homology domain via an unknown mechanism mediates the effects of phosphoinositides on guanine nucleotide exchange activity. Here we tested the possibility that phosphatidylinositol 3-kinase substrates and products control an interaction between the pleckstrin homology domain and the Dbl homology domain, thereby explaining the inhibitory effects of phosphatidylinositol 3-kinase substrates and stimulatory effects of the products. Binding studies using isolated fragments of Vav and Sos indicate phosphatidylinositol 3-kinase substrate promotes the binding of the pleckstrin homology domain to the Dbl homology domain and blocks Rac binding to the DH domain, whereas phosphatidylinositol 3-kinase products disrupt the Dbl homology/pleckstrin homology interactions and permit Rac binding. Additionally, Lck phosphorylation of Vav, a known activating event, reduces the affinities between the Vav Dbl homology and pleckstrin homology domains and permits Rac binding. We also show Vav activation in cells, as monitored by phosphorylation of Vav, Vav association with phosphatidylinositol 3,4,5-trisphosphate, and Vav guanine nucleotide exchange activity, is blocked by the phosphatidylinositol 3-kinase inhibitor wortmannin. These results suggest the molecular mechanisms for activation of Vav and Sos1 require disruption of inhibitory intramolecular interactions involving the pleckstrin homology and Dbl homology domains.

3T3 Cells↗
PubMed2000

Maximum reach envelope for the seated and standing male and female for industrial workstation design.

Maximum reach envelopes for the 5th, 50th and 95th percentile reach lengths of males and females in seated and standing work positions were determined. The use of a computerized potentiometric measurement system permitted functional reach measurement in 15 min for each subject. The measurement system captured reach endpoints in a dynamic mode while the subjects were describing their maximum reach envelopes. An unbiased estimate of the true reach distances was made through a systematic computerized data averaging process. The maximum reach envelope for the standing position was significantly (p<0.05) larger than the corresponding measure in the seated position for both the males and females. The average reach length of the female was 13.5% smaller than that for the corresponding male. Potential applications of this research include designs of industrial workstations, equipment, tools and products.

Adult↗
PubMed2000

Isometric pull and push strengths of paraplegics in the workspace: 1. Strength measurement profiles.

The isometric strength profiles of male and female paraplegics were determined for pull and push strengths in the normal, maximum, and extreme working reach envelopes. A computerized isometric strength measurement system was designed and constructed for the purpose. The strongest pull location was at extreme reach vertically above the shoulder and the strength values for males and females were 473 and 318 newtons (N), respectively. The strongest push location was at maximum reach, at vertical (phi) angle of 45 degrees and at horizontal (theta) angle of 45 degrees for males and at 0 degrees for women and the strength values were 235 and 172 N, respectively. The nature of the strength profiles was found to be similar for both the sexes. The pull and push strengths of the female were 77 and 68% that of the male, respectively.

Adult↗
PubMed2000

Isometric pull and push strengths of paraplegics in the workspace: 2. Statistical analysis of spatial factors.

The effect of reach levels, horizontal angles, and vertical angles on isometric pull and push strengths of male and female was determined. Highly significant increases in men's push strength were found between extreme to maximum reaches, and from extreme to normal reaches. However, for women's push strength, a significant increase was found only between extreme to maximum reach. Significant or highly significant increases were found in men's and women's pull strength between the horizontal angle (theta) sagittal through the active shoulder (90 degrees ) and other angles (0, 45, and 135 degrees ). However, for men's push strength, highly significant increases were found between the horizontal angle 45 degrees from the frontal plane, and other angles. For women's push strength, significant or highly significant increases were found between the horizontal angles 0 degrees and angles of 90 and 135 degrees. For men's and women's pull strength, significant or highly significant increases were found between the vertical angle (phi), 90 degrees, and other angles (-20, 0, and 45 degrees ). Similar increases were found for women's push strength between the 45 degrees angle and other angles. In the design of a workstation for paraplegics that requires pull and push forces, consideration must be given to the spatial factors.

Adult↗
PubMed2000

Evaluation of low back pain risks in a beef skinning operation.

The low back pain risks in a beef skinning operation at a high stand kill floor workstation was evaluated. The increases in compressive forces at lower back (L5/S1) between normal slump (back angle 25 degrees, measured in the sagittal plane) and severe (45 degrees ) and between normal slump and very severe (70 degrees ) bent back postures were 387 N or 28% and 616 N or 45%, respectively. The high spine load coupled with high level of repetition can have a high probability of fatigue failure in the spine structural members. Non-neutral back posture for a large portion of the total work time can be a low back pain risk factor. The videotape analysis showed that the times involved during the task performance for the bent back (more than 25 degrees ) and severe bent back (more than 45 degrees ) were 48.4 and 33.5% of the total cycle time, respectively. The upper limit from OWAS (Ovako Working Posture Analysis System) for bent back posture is 30% of the total cycle time. The bent and twisted back posture (both more than 25 degrees ) time was 10.4% compared to OWAS limit of 5%. This indicated that actions are needed in the near future to alleviate the risk of low back pain. Ergonomics redesign of the workstation was recommended for the operation.

Biomechanical Phenomena↗
PubMed2000

The role of nuclear cap binding protein Cbc1p of yeast in mRNA termination and degradation.

The cyc1-512 mutation in Saccharomyces cerevisiae causes a 90% reduction in the level of iso-1-cytochrome c because of the lack of a proper 3'-end-forming signal, resulting in low levels of eight aberrantly long cyc1-512 mRNAs which differ in length at their 3' termini. cyc1-512 can be suppressed by deletion of either of the nonessential genes CBC1 and CBC2, which encode the CBP80 and CBP20 subunits of the nuclear cap binding complex, respectively, or by deletion of the nonessential gene UPF1, which encodes a major component of the mRNA surveillance complex. The upf1-Delta deletion suppressed the cyc1-512 defect by diminishing degradation of the longer subset of cyc1-512 mRNAs, suggesting that downstream elements or structures occurred in the extended 3' region, similar to the downstream elements exposed by transcripts bearing premature nonsense mutations. On the other hand, suppression of cyc1-512 defects by cbc1-Delta occurred by two different mechanisms. The levels of the shorter cyc1-512 transcripts were enhanced in the cbc1-Delta mutants by promoting 3'-end formation at otherwise-weak sites, whereas the levels of the longer cyc1-512 transcripts, as well as of all mRNAs, were slightly enhanced by diminishing degradation. Furthermore, cbc1-Delta greatly suppressed the degradation of mRNAs and other phenotypes of a rat7-1 strain which is defective in mRNA export. We suggest that Cbc1p defines a novel degradation pathway that acts on mRNAs partially retained in nuclei.

Fungal Proteins↗
PubMed2000

Isolation and characterization of human breast adenocarcinoma cells made resistant to alpha-difluoromethylornithine.

Human breast adenocarcinoma cells MCF-7 were selected for resistance to ornithine decarboxylase (ODC) inhibitor, alpha-difluoromethylornithine (DFMO). Stepwise increments of the concentration of DFMO resulted in selection of MCF-7 cells that were capable of growing in the presence of 1.0 mM DFMO. This capacity was associated with a 10-fold increase in ODC activity and marked enhancement in the synthesis rate of ODC protein as verified by a 2-hr [35S]methionine labeling of cellular proteins followed by immunoprecipitation and SDS-PAGE. The resistant cells had much higher concentration of putrescine, spermidine, and spermine than the control cells. A 25-fold increase in ED50 (effective dose causing 50% inhibition) for the antiproliferative action of DFMO in these resistant cells was observed. The susceptibility of wild-type and resistant cell lines to other inhibitors of the polyamine biosynthetic pathway and adriamycin is also reported.

Adenocarcinoma↗
PubMed2000

Computational fluid dynamic studies of leukocyte adhesion effects on non-Newtonian blood flow through microvessels.

The study of the effect of leukocyte adhesion on blood flow in small vessels is of primary interest to understand the resistance changes in venular microcirculation. Available computational fluid dynamic studies provide information on the effect of leukocyte adhesion when blood is considered as a homogeneous Newtonian fluid. In the present work we aim to understand the effect of leukocyte adhesion on the non-Newtonian Casson fluid flow of blood in small venules; the Casson model represents the effect of red blood cell aggregation. In our model the blood vessel is considered as a circular cylinder and the leukocyte is considered as a truncated spherical protrusion in the inner side of the blood vessel. The cases of single leukocyte adhesion and leukocyte pairs in positions aligned along the same side, and opposite sides of the vessel wall are considered. The Casson fluid parameters are chosen for cat blood and human blood and comparisons are made for the effects of leukocyte adhesion in both species. Numerical simulations demonstrated that for a Casson fluid with hematocrit of 0.4 and flow rate Q = 0.072 nl/s, a single leukocyte increases flow resistance by 5% in a 32 microns diameter and 100 microns long vessel. For a smaller vessel of 18 microns, the flow resistance increases by 15%.

Animals↗
PubMed2000

Cytogenetic toxicity of vincristine.

The anticancer drugs vincristine sulphate (VCR) and cyclophosphamide (CTX) were tested for their cytogenetic effects in the bone marrow cells of Swiss mice. The end points investigated were chromosomal aberrations and mitotic index at 24 hours posttreatment and micronuclei (MN) at 30 hours posttreatment in bone marrow cells of male and female mice after a single intraperitoneal exposure. The doses tested were VCR 0.25, 0.5, and 1.0 mg/kg and CTX 40 mg/kg b.w. of mice. Significant percentages of chromosomal aberrations and significant numbers of micronuclei per thousand polychromatic erythrocytes (PCEs) that were induced were recorded from bone marrow of each of the VCR-treated groups of mice. There were no significant differences between the percentages of dividing cells in the VCR-treated group and the vehicle control groups of mice. Peculiarly, in the chromosomal aberration study, the male mice were found to be more responsive to VCR than the females, and the aberrations per hundred metaphases were found to be decreased when the dose of VCR was increased. The percentage of dividing cells was also higher with the lowest dose of VCR tested. However, there was a dose-dependent, but nonlinear, increase in MN per thousand PCEs. The results were compared with the already available fragmentary and self-contradictory data on the genotoxicity of VCR in mice and in other mammalian test systems.

Animals↗
PubMed1999

Phenotypic reversal of the btn1 defects in yeast by chloroquine: a yeast model for Batten disease.

BTN1 of Saccharomyces cerevisiae encodes an ortholog of CLN3, the human Batten disease gene. We have reported previously that deletion of BTN1, btn1-Delta, resulted in a pH-dependent resistance to D-(-)-threo-2-amino-1-[p-nitrophenyl]-1,3-propanediol (ANP). This phenotype was caused by btn1-Delta strains having an elevated ability to acidify growth medium through an elevated activity of the plasma membrane H(+)-ATPase, resulting from a decreased vacuolar pH during early growth. We have determined that growing btn1-Delta strains in the presence of chloroquine reverses the resistance to ANP, decreases the rate of medium acidification, decreases the activity of plasma membrane H(+)-ATPase, and elevates vacuolar pH. However, an additional effect of this phenotypic reversal is that activity of plasma membrane H(+)-ATPase is decreased further and vacuolar pH is increased further as btn1-Delta strains continue to grow. This phenotypic reversal of btn1-Delta can be considered for developing a therapy for Batten disease.

Chloroquine↗