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Biomedical subjects

M Singh

Publications and source records attributed to M Singh.

At least 379 records · Page 21Linked to original sources
PubMed1999

Monte Carlo simulation of photon scattering in biological tissue models.

Monte Carlo simulation of photon scattering, with and without abnormal tissue placed at various locations in the rectangular, semi-circular and semi-elliptical tissue models, has been carried out. The absorption coefficient of the tissue considered as abnormal is high and its scattering coefficient low compared to that of the control tissue. The placement of the abnormality at various locations within the models affects the transmission and surface emission of photons at various locations. The scattered photons originating from deeper layers make the maximum contribution at farther distances from the beam entry point. The contribution of various layers to photon scattering provides valuable data on variability of internal composition. Introduction.

Models, Anatomic↗
PubMed1999

Evaluation of hemorheological risk factor profile in plasmacytoma patients.

The hemorheological parameters, plasma viscosity, hematocrit, erythrocyte aggregation and deformability of blood samples obtained from plasmacytoma patients are measured by capillary viscometer, microcentrifuge, Myrenne aggregometer, and filtrometer, respectively. These parameters are significantly altered in patients compared to those of normal subjects. The plasma viscosity and erythrocyte aggregation are increased, whereas erythrocyte deformability and hematocrit are decreased. Based on these parameters the hemorheological risk factor profiles are developed and by their overall risk factors, the patients with low and high risk factors are identified.

Blood Viscosity↗
PubMed1999

Influence of local anaesthetics on the aggregation and deformability of erythrocytes.

The effects of two local anaesthetics, procain and tetracaine, on erythrocyte aggregation and deformability under in vitro conditions are analysed. Fresh blood samples from healthy volunteers are obtained by venepuncture in test tubes containing heparin as anticoagulant. After centrifugation, plasma and cells are separated and buffy coat is discarded. The erythrocyte suspensions for aggregation and deformability measurement are prepared at hematocrit 34 and at 8% in buffered medium containing glucose and albumin, respectively. A part of the suspending medium in these samples is replaced by an equal volume of anaesthetic solution to obtain its final concentration ranging from 0.001 to 1 g/l. The results show that with the increase of anaesthetic concentration the deformability index and the aggregation index are reduced. The change in erythrocyte shape may contribute to the decrease in aggregation. The combined effect of these parameters may affect blood flow under varied experimental conditions.

Anesthetics, Local↗
PubMed1998

Reversal of a novel multidrug resistance mechanism in human colon carcinoma cells by fumitremorgin C.

We selected a human colon carcinoma cell line in increasing concentrations of mitoxantrone to obtain a resistant subline, S1-M1-3.2, with the following characteristics: profound resistance to mitoxantrone; significant cross-resistance to doxorubicin, bisantrene, and topotecan; and very low levels of resistance to Taxol, vinblastine, colchicine, and camptothecin. This multidrug resistance (MDR) phenotype, which was not reversed by verapamil or another potent P-glycoprotein (Pgp) inhibitor, CL 329,753, was dependent, in part, upon an energy-dependent drug efflux mechanism. Pgp and the multidrug resistance protein (MRP) were not elevated in the resistant cells relative to the drug-sensitive parent, suggesting that resistance was mediated by a novel pathway of drug transport. A cell-based screen with S1-M1-3.2 cells was used to identify agents capable of circumventing this non-Pgp, non-MRP MDR. One of the active agents identified was a mycotoxin, fumitremorgin C. This molecule was extremely effective in reversing resistance to mitoxantrone, doxorubicin, and topotecan in multidrug-selected cell lines showing this novel phenotype. Reversal of resistance was associated with an increase in drug accumulation. The compound did not reverse drug resistance in cells with elevated expression of Pgp or MRP. We suggest that fumitremorgin C is a highly selective chemosensitizing agent for the resistance pathway we have identified and can be used as a specific pharmacological probe to distinguish between the diverse resistance mechanisms that occur in the MDR cell.

ATP Binding Cassette Transporter, Subfamily B↗
PubMed1998

The preparation and characterization of polymeric antigen delivery systems for oral administration.

Although polymeric delivery systems are well established for the oral administration of conventional drugs, they have not yet been commercially developed for vaccine delivery. The problems inherent with the oral route of delivery, including low pH, gastric enzymes, rapid transit and poor absorption of large molecules, has made the goal of oral delivery of antigens very challenging. Nevertheless, several polymeric delivery systems for the oral administration of vaccines are currently being evaluated, including microencapsulation in poly(lactide-co-glycolides), alginates, polyanhydrides, starch, polymethacrylates, polyamino acids and enteric coating polymers. These approaches are designed to protect the antigen in the gut, to target the antigen to the gut-associated lymphoid tissue, or to increase the residence time of the antigen in the gut through bioadhesion. Each of these approaches is discussed in relation to antigen encapsulation and integrity, process reproducibility, ease of preparation and encapsulation efficiency. Potential problems associated with the scale-up of these approaches are also briefly addressed. Of particular relevance are the prospects for the application of these formulation processes for commercial development.

Journal Article↗
PubMed1998

Antimutagenicity of hydrolyzable tannins from Terminalia chebula in Salmonella typhimurium.

A tannin fraction (TC-E) from the dried fruit pulp of Terminalia chebula was obtained by successfully extracting with 95% ethyl alcohol and ethyl acetate. TC-E was subjected to silica gel chromatography which yielded four fractions, viz., TC-EI, TC-EII, TC-EIII and TC-EIV. Thin layer chromatography (TLC) and 13C-NMR revealed that TC-EI was gallic acid (GA) derivative while the other fractions were tannin in nature. TC-E and its fractions were evaluated for their antimutagenic potential against two direct-acting mutagens, 4-nitro-o-phenylenediamine (NPD) and 4-nitroquinoline-N-oxide (4NQNO), and S9-dependent mutagen, 2-aminofluorene (2AF) in TA98 and TA100 strains of Salmonella typhimurium. The study revealed that the extract (TC-E) and its fractions were highly significant against S9-dependent mutagen, 2AF. The effect was found to be more or less corresponding with the nature of the fractions, as the monomeric TC-EI (a GA derivative) was least effective as compared to other fractions which were oligomeric, and the order of their effectiveness as per their IbD50 value being TC-EIV (8.9 micrograms)>TC-EIII (17.8 micrograms)>TC-EII (45 micrograms)>TC-EI (320 micrograms) in TA98; TC-EIV being 40 times more effective than TC-EI in inhibiting his+ revertants. A similar effect was noticed in TA100 too, where TC-EI was the least effective and TC-EII had the maximum effect. A similar result was noticed when the antimutagenicity of GA (a monomeric) was compared with tannic acid (TA, an oligomeric). However, chebula tannins were found to be partly effective against NPD but not at all effective against 4NQNO.

Antimutagenic Agents↗
PubMed1998

A novel method of purifying lung surfactant proteins A and D from the lung lavage of alveolar proteinosis patients and from pooled amniotic fluid.

A simple procedure has been developed for the purification of the surfactant proteins SP-A and SP-D from lung lavage of patients with alveolar proteinosis. The SP-D is purified by fractionation of the supernatant obtained after spinning the lavage at 10000 X g for 40 min, while the bulk of the SP-A is purified by fractionation of the pellet. The supernatant is applied to a maltosyl-agarose column and the bound SP-D is specifically eluted using MnCl2. The pellet is solubilised in 6 M urea and, following renaturation, the solubilised proteins are applied to maltosyl-agarose and SP-A eluted using a gradient of EDTA. Both SP-A and SP-D are further purified by gel-filtration on Superose-6. This procedure has also been used to prepare successfully human SP-A and SP-D from amniotic fluid and may be generally applicable to the isolation of these surfactant proteins from lung washings obtained from other species.

Amniotic Fluid↗
PubMed1998

Poly(lactide-co-glycolide) microparticles for the development of single-dose controlled-release vaccines.

With the exception of the provision of clean water supplies, vaccination remains the most successful public health intervention strategy for the control of infectious diseases. However, the logistics of delivering at least two to three doses of vaccines to achieve protective immunity are complex and compliance is frequently inadequate, particularly in developing countries. In addition, newly developed purified subunit and synthetic vaccines are often poorly immunogenic and need to be administered with potent vaccine adjuvants. Microparticles prepared from the biodegradable and biocompatible polymers, the poly(lactide-co-glycolides) or (PLG), have been shown to be effective adjuvants for a number of antigens. Moreover, PLG microparticles can control the rate of release of entrapped antigens and therefore, offer potential for the development of single-dose vaccines. To prepare single-dose vaccines, microparticles with different antigen release rates may be combined as a single formulation to mimic the timing of the administration of booster doses of vaccine. If necessary, adjuvants may also be entrapped within the microparticles or, alternatively, they may be co-administered. The major problems which may restrict the development of microparticles as single-dose vaccines include the instability of vaccine antigens during microencapsulation, during storage of the microparticles and during hydration of the microparticles following in vivo administration. In the present review, we discuss the adjuvant effect of PLG microparticles, and also their potential for the development of single-dose vaccines through the use of controlled-release technology.

Journal Article↗
PubMed1998

Host vector system for high-level expression and purification of recombinant, enzymatically active alanine dehydrogenase of Mycobacterium tuberculosis.

The 40-kDa antigen of M. tuberculosis, which is an alanine dehydrogenase, is a species-specific antigen that is potentially useful for strain identification. Large quantities of the purified protein are required for immunological, as well as for detailed biochemical and structural, characterization. The AlaDH gene was cloned by PCR from H37Rv (virulent) and H37Ra (partially attenuated) strains of M. tuberculosis, and their DNA sequence was determined. A host-vector system suitable for the production of sufficient quantities of the recombinant AlaDH antigen was developed. The AlaDH gene was expressed under the control of strong, transcriptional (bacteriophage pLpR) and translational (atpE) signals. High-level expression of soluble AlaDH was obtained using the recombinant E. coli K-12 strain CAG629 [pMSK12], which is deficient in Lon protease and the heat-shock response. A simple two-step procedure for the rapid purification of the recombinant protein was developed. The protein was purified to near homogeneity, and the purified AlaDH showed a specific enzyme activity comparable to the native protein isolated from M. tuberculosis. In addition, the product showed an expected amino acid sequence and reacted strongly to the 40-kDa (AlaDH)-specific mAb HBT-10. Furthermore, the epitope of the mAb HBT-10 was mapped to a 12-amino-acid region. Contrary to the published results, we show that the AlaDH and the PNT (pyridine nucleotide transhydrogenase) of M. tuberculosis do not share common epitopes reacting to the species-specific mAb HBT-10. The availability of highly purified AlaDH should now enable a detailed biochemical and structural characterization of this important enzyme of M. tuberculosis.

Alanine Dehydrogenase↗
PubMed1998

Computational learning reveals coiled coil-like motifs in histidine kinase linker domains.

The recent rapid growth of protein sequence databases is outpacing the capacity of researchers to biochemically and structurally characterize new proteins. Accordingly, new methods for recognition of motifs and homologies in protein primary sequences may be useful in determining how these proteins might function. We have applied such a method, an iterative learning algorithm, to analyze possible coiled coil domains in histidine kinase receptors. The potential coiled coils have not yet been structurally characterized in any histidine kinase, and they appear outside previously noted kinase homology regions. The learning algorithm uses a combination of established sequence patterns in known coiled coil proteins and histidine kinase sequence data to learn to recognize efficiently this coiled coil-like motif in the histidine kinases. The common appearance of the structural motif in a functionally important part of the receptors suggests hypotheses for kinase regulation and signal transduction.

Algorithms↗
PubMed1998

Sequence analysis of the tryparedoxin peroxidase gene from Crithidia fasciculata and its functional expression in Escherichia coli.

Tryparedoxin peroxidase from Crithidia fasciculata is an essential component of the trypanothione-dependent hydroperoxide metabolism in the trypanosomatids (Nogoceke, E., Gommel, D. U., Kiebeta, M., Kalisz, H. M., and Flohé, L. (1997) Biol. Chem. 378, 827-836). The tryparedoxin peroxidase gene and its flanking regions have been isolated and sequenced from a C. fasciculata genomic DNA library. It consists of an open reading frame of 564 base pairs encoding a protein of 188 amino acid residues. The gene, modified to encode 6 additional histidine residues, was expressed in Escherichia coli and the recombinant protein was purified to homogeneity by metal chelating chromatography. Recombinant tryparedoxin peroxidase has a subunit molecular mass of 21884 +/- 22 and contains two isoforms of pI 6.2 and 6.3. It exhibits a kinetic pattern identical to that of the authentic tryparedoxin peroxidase and has a similar specific activity of 2.51 units mg-1. The enzyme unequivocally belongs to the peroxiredoxin family of proteins, whose members have been found in all phyla. A phylogenetic tree comprising 47 protein and DNA sequences showed tryparedoxin peroxidase and a homologous Trypanosoma brucei sequence to form a distinct molecular clade. The consensus sequence: xnAx5-6Fx9Gx3Vx2Fx1Px2Fx1FVCPTEx21Sx1Dx7Wx16-19Dx15- 16Gx3Rx2Fx2Dx27Ax 1Qx4-11Cx1-3Wxn was demonstrated by alignment of the sequences of tryparedoxin peroxidase and 8 other peroxiredoxins with established peroxidase function.

Amino Acid Sequence↗
PubMed1998

Role of monensin PLGA polymer nanoparticles and liposomes as potentiator of ricin A immunotoxins in vitro.

Monensin is a carboxylic ionophore which can potentiate the activity of ricin based immunotoxins (IT) in vitro and in vivo against a variety of human tumours. Monensin was encapsulated into nanoparticles (NP) by using biodegradable poly(DL-lactide-co-glycolide) (PLGA, 50:50). The NP were prepared by modified emulsification-solvent evaporation method. High shear homogenization followed by simultaneous stirring and bath sonication were used for preparing NP. The size of NP was determined by photon correlation spectroscopy using a BI 90 particle sizer (Brookhaven Instruments). The average size of NP could be decreased from 567 nm to 163 nm by increasing the concentration of polyvinyl alcohol from 10% to 100% of PLGA. The NP were spherical in shape as observed by Atomic Force Microscopy. The concentration of monensin in the NP was analyzed by HPLC and the entrapment efficiency was found to be more than 12%. The zeta potential of NP was -25.8 (+/- 1.3) mv, which did not change significantly after resuspension of the freeze dried sample. The NP were tested against HL-60 and HT-29 human tumour cell lines in vitro. Monensin NP potentiated the activity of IT by 40 to 50 times against these cell lines. There was however, no difference between the NP and liposomes for their potentiating affect of IT against the two tumour cell lines.

Humans↗
PubMed1998

Fine-needle aspiration biopsy of colonic masses.

Between 1989 and 1996, fine-needle aspiration biopsy was performed in 22 patients with palpable colonic masses. In all of these patients colonoscopic examination was either not possible or could not be carried out successfully. The results of aspiration biopsy were confirmed by surgery and histopathological examination of the tissue. Aspiration biopsy correctly diagnosed all except one case. It identified all cases of colonic cancer. One patient with colonic tuberculosis was also diagnosed correctly. However, in another patient with colonic tuberculosis, aspiration biopsy showed only nonspecific changes in the form of inflammatory and epithelial cells. There were no false-positive results or complications from the procedure. It is concluded that fine-needle aspiration biopsy is a simple, rapid, and accurate method of diagnosing palpable colonic masses in patients in whom a colonoscopy is not possible or cannot be performed satisfactorily.

Adenocarcinoma↗
PubMed1998

Volume localized in vivo proton MR spectroscopy of breast carcinoma: variation of water-fat ratio in patients receiving chemotherapy.

Results are reported on in vivo volume localized proton magnetic resonance spectroscopy (MRS) of patients (n = 44) suffering from carcinoma of the breast, using a bilateral breast surface coil. Localized proton MR spectra of the unaffected contralateral breast of these patients are dominated by resonances arising from fat and are similar to the breast tissue from normal volunteers (controls, n = 13), while in the malignant breast tissues the water resonance dominates. On the other hand, the water suppressed proton MR spectra of malignant breast tissue reveal several metabolites of low concentration including the choline peak around 3.2 ppm and other resonances attributable to purine and pyrimidine nucleotides, in the 8.5 ppm region. Elevated water- fat (W-F) ratios are measured in the malignant tissues, compared with the normal breast tissue of controls and from the contralateral unaffected breast tissue of the patients (n = 11). In the case of patients receiving chemotherapy resulting in the reduction of primary tumor size, the W-F ratio shows a statistically significant (P < 0.01) decrease compared with the pre-therapy value, thus providing a non-invasive indicator of favourable clinical outcome of neoadjuvant chemotherapy for locally advanced breast cancer. The method provides the potential for non-invasively monitoring and assessing the response of breast cancer to neoadjuvant chemotherapy.

Adult↗
PubMed1998

CD8(+), radiosensitive T cells of parental origin, oppose cells capable of down-regulating cytotoxicity in murine acute lethal graft-versus-host disease.

Murine graft-versus-host (GVH) disease takes two forms depending upon the parental/F1 strain combination employed. Anemia, lymphopenia, hypogammaglobulinemia, profound anti-F1 cytotoxicity, and the loss of cytotoxic potential against third party alloantigen is seen in acute lethal GVH disease. In contrast to this, in chronic GVH disease there is polyclonal B cell activation, auto-antibody production, no anti-F1 cytotoxicity, and retained cytotoxicity against allotargets. We have previously reported that this marked disparity in disease expression results from a radiosensitive host veto cell which protects the F1 mouse from parental anti-F1 cytotoxicity in mice undergoing CGVH disease. This cell could be induced in vitro or in vivo in CGVH disease. Using an in vitro system, we now demonstrate that a CD4(+), radiation-sensitive, T cell does emerge in acute lethal GVH disease which is capable of down-regulating cytotoxicity. The cell does not appear to be a veto cell in that it attenuates cytotoxicity directed against nonself alloantigen. The function of this cell does not appear to be influenced by minor lymphocyte stimulatory gene products. We further report that, in ALGVH disease, regulation by this cell is not readily apparent due to the emergence of a CD8(+) T cell of parental (B6) origin, which opposes its action.

Acute Disease↗
PubMed1998

Rotational Analysis of the 0-0 Band of the a3Pi-X1Sigma+ Transition of AlCl

The 0-0 band of the a3Pi-X1Sigma+ transition of AlCl at 4073.6 A has been photographed at high resolution and an unambiguous analysis of its rotational structure is presented. Precise values of the rotational constants of the a3Pi state have been determined. Copyright 1998 Academic Press. Copyright 1998Academic Press

Journal Article↗
PubMed1998

Genomic polymorphism among Blastocystis hominis strains and development of subtype-specific diagnostic primers.

Genomic polymorphisms among nine strains of Blastocystis hominis were examined by random amplified polymorphic DNA (RAPD) using four different arbitrary polymerase chain reaction (PCR) primers. Based on the RAPD patterns, nine strains were classified into three groups. The specific primers designed from the unique bands yielded a single band from within each same group, but did not amplify between all the groups examined. Specificity of these diagnostic primers was tested against several common intestinal parasites and a yeast, and no amplification was confirmed. Since the current criteria indicates that Blastocystis organisms isolated from humans are designated as B. hominis, the authors propose to classify several subtypes among B. hominis groups based on the difference of genomic DNA using three diagnostic primers.

Animals↗