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Biomedical subjects

P Bhatnagar

Publications and source records attributed to P Bhatnagar.

At least 19 recordsLinked to original sources

Evidence-based practice in the utilization of knee radiographs--a survey of all members of the British Orthopaedic Association.

There has been a debate amongst orthopaedic surgeons about which plain radiographs are required in common knee conditions and this is a 5-year review assessing the influence of recent publications on the use of radiographs. A postal survey of all members of the British Orthopaedic Association was performed. The response rate was 60% (990/1,650). There was a broad agreement on the use of anteroposterior and lateral radiographs. The use of the posteroanterior (PA) weight-bearing radiograph has increased from 82 to 86%. The use of skyline view for suspected arthritis and after knee replacement has increased from 23 to 41%. There is still disparity amongst orthopaedic surgeons regarding the knee flexion angle for skyline and weight-bearing views. After review of the literature we recommend that a single skyline view in 30 degrees flexion is adequate and should be a standard investigation in knee disorders. We also stress the importance of weight-bearing PA radiograph in 30 degrees knee flexion for adequate assessment in all patients with suspected arthritis.

Arthrography↗

Assessment of organochlorine pesticide residue levels in dairy milk and buffalo milk from Jaipur City, Rajasthan, India.

The widespread application of pesticides in agriculture, public health, and industry and in and around the home can result in the accumulation of pesticides in the environment. Therefore, a survey has been conducted during 1993-1996 to investigate the magnitude of contamination of bovine milk with organochlorine pesticide (OCP) residues from Jaipur City, Rajasthan, India. Milk samples, i.e., dairy (toned and whole) and buffalo milk, were collected seasonally, and pesticide residues were assessed using a gas chromatograph (GC) with an electron capture detector (ECD). The results indicate that all the milk samples were contaminated with dichlorodiphenyltrichloroethane (DDT) and its metabolites (DDE and p,p'-dichlorodiphenyldichloroethane [DDD]), isomers of hexachlorocyclohexane (HCH; alpha, beta, and gamma), heptachlor and its epoxide, and aldrin. Seasonal variations of these pesticide residue levels were also observed in all the milk samples. Samples collected during winter season were found to contain higher residue levels as compared to other seasons.

Animals↗

Transgenic manipulation of the metabolism of polyamines in poplar cells.

The metabolism of polyamines (putrescine, spermidine, and spermine) has become the target of genetic manipulation because of their significance in plant development and possibly stress tolerance. We studied the polyamine metabolism in non-transgenic (NT) and transgenic cells of poplar (Populus nigra x maximowiczii) expressing a mouse Orn decarboxylase (odc) cDNA. The transgenic cells showed elevated levels of mouse ODC enzyme activity, severalfold higher amounts of putrescine, a small increase in spermidine, and a small reduction in spermine as compared with NT cells. The conversion of labeled ornithine (Orn) into putrescine was significantly higher in the transgenic than the NT cells. Whereas exogenously supplied Orn caused an increase in cellular putrescine in both cell lines, arginine at high concentrations was inhibitory to putrescine accumulation. The addition of urea and glutamine had no effect on polyamines in either of the cell lines. Inhibition of glutamine synthetase by methionine sulfoximine led to a substantial reduction in putrescine and spermidine in both cell lines. The results show that: (a) Transgenic expression of a heterologous odc gene can be used to modulate putrescine metabolism in plant cells, (b) accumulation of putrescine in high amounts does not affect the native arginine decarboxylase activity, (c) Orn biosynthesis occurs primarily from glutamine/glutamate and not from catabolic breakdown of arginine, (d) Orn biosynthesis may become a limiting factor for putrescine production in the odc transgenic cells, and (e) assimilation of nitrogen into glutamine keeps pace with an increased demand for its use for putrescine production.

Animals↗

Inhibition of enriched stem cells in vivo and in vitro by the hemoregulatory peptide SK&F108636.

Replacement of the labile sulfhydryl group (-SH) of the hemoregulatory peptide monomer pyroGluGluAspCysLys (HP5b) with an isosteric methylene group yields a chemically stable compound, SK&F108636. In this study, we describe the effects of SK&F108636 on highly enriched Lin-Sca1+ hematopoietic stem cells. SK&F108636 significantly reduced the fraction of cycling progenitor cells, granulocyte macrophage colony-forming cells (GM-CFC), in vitro and in vivo. There was no effect on GM-CFC or Mix-CFC colony formation. SK&F108636 significantly inhibited proliferation of high proliferative potential (HPP)-CFC in semisolid agar cultures stimulated by stem cell factor + interleukin 3 (IL-3) + IL-1, but had no effect in cultures stimulated with M-CSF + IL-3 + IL-1. SK&F108636 was shown to act directly on the stem cells since SK&F108636 inhibited proliferation of Lin-Sca1+ cells in single cell assays. Administration of SK&F108636 to lethally irradiated mice transplanted with 2000 Lin-Sca1+ cells significantly inhibited proliferation/differentiation of cells developing into colony forming units-spleen (CFU-S) (preCFU-S) and the reconstitution of HPP-CFC and GM-CFC. There was no effect of SK&F108636 on CFU-S colony formation or mature cell regeneration in bone marrow, spleen and blood. Hence, the hemoregulatory peptide monomer SK&F108636 is a potent primitive stem cell inhibitor in vivo and in vitro. Inhibition of stem cell proliferation by small specific inhibitors may protect hematopoiesis from myelotoxic side effects during chemotherapy treatment.

Animals↗

Zinc is a possible toxic contaminant of silicone oil in microdrop cultures of preimplantation mouse embryos.

A batch of silicone oil (dimethylpolysiloxane) is described which had differential effects on the development of 1- and 2-cell preimplantation mouse embryos in vitro when used as a microdrop overlay over two culture media: CZB and KSOM. A high rate of development into blastocysts was observed when using CZB medium; in contrast, development was strongly inhibited when KSOM was used. Other batches of silicone oil or paraffin oil permitted development from the zygote to the blastocyst of an outbred strain of mouse without arrest at the 2-cell stage. Our results show that the higher concentrations of ethylenediaminetetraacetic acid (EDTA) and bovine serum albumin (BSA) in CZB medium, in comparison with KSOM, protect against the toxic component in the oil. Observations also gave circumstantial evidence that the toxic component in the oil is zinc. The beneficial effect of including EDTA in a medium is usually attributed to its chelating toxic metals introduced as impurities in other components of the medium. Our results now show that EDTA also protects against impurities in the oil overlay.

Animals↗

CSF-1 and mouse preimplantation development in vitro.

The effects of macrophage colony stimulating factor on the development of the zygote to the blastocyst stage of an outbred strain of mouse have been studied in KSOM, an improved medium that supports a high rate of in vitro development. Macrophage colony stimulating factor accelerates the formation of the blastocyst cavity by day 4 (96 hours post-hCG). It also increases overall embryonic cell number through a differential increase in the number of trophoblast cells, with no significant effect on the number of inner cell mass cells. By day 5 of culture (120 hours post-hCG), colony stimulating factor-treated embryos have about 20 more trophoblast cells than control embryos, an increase of about 30 percent of the total number of cells in a control blastocyst. The maximum response of embryos was obtained at a concentration around 540 U ml-1 colony stimulating factor (identical to 918 Stanley units ml-1), and the cytokine can produce the same effects even if it is present in the medium for only part of the culture period. This in vitro stimulation of preimplantation development with macrophage colony stimulating factor is compatible with continued normal fetal development in vivo.

Animals↗

Interobserver agreement in the diagnosis of cervical smears.

The present study was carried out to reveal the magnitude of individual variation in the diagnosis of Pap smears between two cytoscreeners and their compatability with cytopathologists and subsequent final diagnostic comparison with biopsy in 1,17411 cervical smears collected from different hospitals of Delhi during ten-year period. Smears diagnosed as dysplasia at initial level by any one of the cytoscreeners were screened by cytopathologists for confirmation of diagnosis. An overall agreement of 94.9 percent was observed between two screens. 79.5 percent was agreement between screeners and cytopathologists. An agreement between cytology and histology in the diagnosis of dysplasia and malignancy were found to be 61.9 percent and dysplasia and malignancy were found to be 61.9 percent and 40.1 percent respectively. From the above study, it was observed that consistency among the two screeners was fairly acceptable. Keeping this observation in view, we can possibly practice two tier screening in place of three.

Female↗

Mapping of neutralizing epitopes and the receptor binding site of human interleukin 1 beta.

Antibodies to synthetic peptides of human interleukin 1 beta (IL-1 beta) and to recombinant human IL-1 beta were used to identify epitopes of IL-1 beta associated with the neutralization of its biological activity. Analysis of antisera raised to 17 synthetic peptides derived from the mature IL-1 beta sequence showed that five regions (residues 6-15, 49-80, 58-80, 92-101, and 120-133) were both immunoprecipitating and neutralizing. Using a hexamer epitope mapping method, comparison of the regions recognized by four neutralizing rabbit antisera with those recognized by a rabbit antiserum raised to denatured IL-1 beta suggested two further neutralizing epitopes, residues 39-48 and 83-95. Finally, a neutralizing monoclonal antibody was shown to bind to the peptides 6-11 and 87-95 by peptide binding and mutagenesis. All of these regions appear predominantly on one face of IL-1 beta. The effect of mutations in residues 4-11 and 88-97, which lie within this face, on receptor binding and biological activity was determined. Most of the mutations tested affected both receptor binding and activity, whereas mutations in another face of IL-1 beta (residues 74-80) had no effect. Purification of two of the mutants with reduced bioactivity and receptor binding and analysis by two-dimensional NMR indicated no gross changes in tertiary structure. A third mutant had reduced bioactivity in two different bioassays but no change in receptor binding. Although two-dimensional NMR revealed no gross changes in conformation, small changes did occur at a site distal from that mutated. The data are consistent with other epitope mapping and receptor binding mutagenesis data and suggest that the neutralizing antibodies and receptor recognize different but overlapping regions of IL-1 beta.

Amino Acid Sequence↗

Inflammatory lesions of the breast: diagnosis by fine needle aspiration.

Amongst 1061 breast lesions diagnosed by fine needle aspiration (FNA) over a period of 6 years (1985-1990), 128 were reported to be showing changes consistent with an inflammatory lesion. On review, the cytodiagnosis was found to be inaccurate in 31 cases. The cytological features of the 97 cases that were correctly reported are described in this report. The cytological diagnoses issued in these 97 cases were acute mastitis or breast abscess (57 cases) and tuberculous mastitis (30 cases). Non-specific chronic mastitis and miscellaneous conditions accounted for four and six cases respectively. Acid fast bacilli (AFB) were demonstrated in 28.0% of tuberculous mastitis cases and 10.0% of those diagnosed as acute mastitis or breast abscess. FNA cytology was found to be useful for the diagnosis of inflammatory lesions of breast and their classification, as only five out of 57 cases of acute mastitis/breast abscess and one out of 30 tuberculous mastitis cases were suspected on clinical grounds.

Abscess↗

A teratogenic study of carbaryl in Swiss albino mice.

To study the teratogenicity of carbaryl, groups of 10 pregnant mice were dosed, by gavage, with 0, 100, 150 or 200 mg carbaryl/kg body weight, in corn oil, on day 8 or day 12 of pregnancy or daily (as daily doses) from day 6 to day 15. The two higher doses were toxic to both dams and foetuses regardless of the timing of treatment. Treated dams generally showed reduced weight gains but total weight gain was significantly reduced only for dams given 200 mg carbaryl on day 8 or day 12 of gestation. Maternal mortality was increased in most groups given 150 or 200 mg carbaryl. Carbaryl treatment tended to reduce litter size, to increase the percentage of resorbed foetuses, and to reduce foetal weight. There were increased incidences of open eye, of certain visceral abnormalities, and of reduced ossification in virtually all treated groups. The variety of abnormalities in treated foetuses reflected the dysmorphogenic potential of the pesticide. More aberrations were seen in foetuses from dams treated throughout organogenesis than in those from dams given a single dose.

Abnormalities, Drug-Induced↗

Antiserum to a novel peptide sequence reacts selectively with epithelial subpopulations.

A synthetic peptide corresponding to a novel protein sequence isolated from bovine kidney was used to immunize rabbits. When applied to Western blots of bovine kidney extracts, antiserum to this peptide recognizes proteins with molecular weights of 23 and 18 KD. Immunohistochemical examination of a variety of bovine and rat tissues with this antiserum revealed a unique distribution of immunoreactivity with the intermediate layers of a variety of stratified epithelia, in addition to renal glomeruli. The pattern of reactivity differed from previously described epithelial markers such as cytokeratins. These results indicate that this antiserum may be useful as a tool for the identification of cells of the intermediate layer of stratified epithelia and, as such, may aid in the study of this differentiating/proliferating tissue compartment.

Amino Acid Sequence↗

Identification and characterization of calcitonin gene-related peptide receptors in porcine renal medullary membranes.

Membranes prepared from the medullary region of the porcine kidney displayed high affinity, high density (Kd, 0.12 nM; binding capacity, 127 fmol/mg protein) receptors for calcitonin gene-related peptide (CGRP). Human CGRP (hCGRP), rat CGRP (rCGRP), and the hCGRP analog [hCGRP-(8-37)] competed for the binding of [125I]hCGRP, whereas salmon calcitonin (sCT) and CGRP-(22-37) were very weak in displacing [125I]hCGRP binding. In accordance with these binding data, CGRP stimulated adenylate cyclase activity in these membrane preparations in a concentration-dependent manner, with an EC50 similar to that of the Kd for binding. In the same preparations, sCT was ineffective in stimulating adenylate cyclase activity, suggesting that porcine kidney medullary membranes possess receptors specific for CGRP. Further hCGRP-(8-37), a CGRP antagonist, inhibited CGRP-stimulated adenylate cyclase activity in a competitive manner. Covalent cross-linking of [125I]hCGRP to these membranes resulted in the specific labeling of one major band at approximately 30,000 mol wt and two minor bands at about 58,000 and 78,000 mol wt. The presence of CGRP receptors and their coupling to adenylate cyclase suggest a role for CGRP in kidney function, such as local regulation of the microcirculation, electrolyte transport, or water homeostasis in the porcine kidney.

Adenylyl Cyclases↗

Clinical ventilatory functions and immunological studies in bronchial asthma.

Clinical characteristics, ventilatory functions and serum immunoglobulins (IgG, IgM and IgA) were assessed in 30 asthmatic patients. Sixty per cent patients belonged to early onset group ie, had onset of disease before 30 years of age. Seasonal asthmatics formed the largest group (56.6%) followed by perennial (26.6%) and irregular (16.6%) type. On the basis of ventilatory function tests, maximum number of cases (15) had asthma of moderate severity, followed by mild (12 patients) and severe forms (3 patients). Mean serum levels (mg%) of IgG (1631.93 +/- 270.71) and IgM (233.82 +/- 86.93) were significantly elevated in asthmatics as compared to controls. Higher serum IgA levels were observed in bronchial asthma patients (257.58 +/- 81.06) as compared to controls (225.69 +/- 49.75), but this rise was not statistically significant. Duration, periodicity of asthmatic attacks and the severity had no influence on the levels of the above three mentioned immunoglobulins, but it was observed that patients who had associated other allergies and positive family history of atopic disorders exhibited higher serum IgA and IgM levels, compared to those who had no such history. This difference was however not seen in serum IgG levels.

Adult↗