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Biomedical subjects

M Mathur

Publications and source records attributed to M Mathur.

At least 91 records · Page 5Linked to original sources

Nosocomial outbreak of Salmonella typhimurium infection in a nursery intensive care unit (NICU) and paediatric ward.

A nosocomial outbreak of multidrug resistant Salmonella typhimurium in a Nursery Intensive Care Unit (NICU) and Paediatric ward is reported. Eight (16.6%) out of a total of 48 babies taken ill during this outbreak expired. Clinical manifestations included diarrhoea and fever. The organism was isolated from stool samples/rectal swabs of all the 48 cases and blood of 6 cases. An investigation was undertaken to trace the source of infection. The organism was isolated in pure cultures from three suction machines of NICU. The epidemic was immediately controlled with identification of the source but the question of how the suction machines were infected by this organism remained unsolved.

Child, Preschool↗

Interactions of the cytoplasmic domain of the desmosomal cadherin Dsg1 with plakoglobin.

Dsg1 is a 165-kDa glycoprotein component of suprabasal epidermal desmosomes and the prototype of a subset of the cadherin superfamily of cell-cell adhesion proteins known as desmogleins. The adhesive function of classical cadherins is known to be dependent upon their association with cytoplasmic components called catenins. In the case of desmogleins, a single interaction has been described with a protein called plakoglobin that is found in desmosomal plaques, adherens junctions, and the cytosol. Several proteins with homology to plakoglobin have been described that regulate junction assembly and implement morphoregulatory signals. To address the functional significance of plakoglobin-desmoglein interaction, we have mapped the sequences of Dsg1 that are crucial for this association by using blot overlay techniques. By examining the binding of plakoglobin to a deletion series of the Dsg1 cytoplasmic domain expressed as fusion proteins, we have defined a 19-amino acid sequence that is important for association. This region of Dsg1 sequence shows significant similarity to the catenin-binding domain of classical cadherins, suggesting a common mechanism for the association of plakoglobin with desmosomes and adherens junctions.

Amino Acid Sequence↗

Structure of DSG1, the bovine desmosomal cadherin gene encoding the pemphigus foliaceus antigen. Evidence of polymorphism.

The cadherin superfamily of calcium-dependent cell-cell adhesion and recognition proteins can be categorized into a number of subsets on the basis of the distinct cytoplasmic sequences of their members. Currently these families include classical cadherins, desmogleins, desmocollins, protocadherins, and the products of the Drosophila genes FAT and Dachsous. Dsg1, the prototype of the desmoglein family, is a major component of epidermal desmosomes and the antigenic target of antibodies found in the sera of patients with the blistering disease, pemphigus foliaceus. In this study, we determined the organization of the bovine DSG1 gene. This gene consists of 15 exons distributed over > 37.5 kilobases of genomic DNA. A comparison of DSG1 with genes encoding classical cadherins revealed a striking conservation of exon boundaries in regions encoding the ectodomain and to a more limited extent among those encoding the cytoplasmic domain. Polymorphism was found in a sequence of DSG1 encoding protein proximal to the external face of the plasma membrane. This region is topologically equivalent to a domain of classical cadherins that harbors epitopes recognized by adhesion-disrupting antibodies. We discuss these results with regard to the evolution of the cadherin superfamily and their implications for the definition of pemphigus epitopes.

Animals↗

Duplications of the alimentary tract: clues to the missing links.

This is a review of the past 10 years' experience with duplications of the alimentary tract at the All India Institute of Medical Sciences, New Delhi, during which 14 infants and children with a total of 15 duplications were treated. Esophageal duplications represented the majority of cases (55%). Certain unexplained associations in this malformation led the authors to perform (1) detailed histological studies of the duplication cysts to seek explanation for the high incidence of gastric mucosa in their walls and its presence in duplications even in the distal parts of the gut, and (2) myelography and postmyelography computerized axial tomography scans to substantiate the spinal pathology. Three of four myelograms in patients with esophageal duplications showed an intraspinal pathology, and all three had vertebral defects. Histological evaluation of the cyst wall showed that gastric mucosa was the most common ectopic tissue in the walls of the cysts, and the tissues were in various primitive stages of differentiation. On the basis of these data, the authors hypothesize that (1) the embryonic endoderm of the entire alimentary tract has the potential to develop into gastric mucosa at the time of inception of duplications, which explains the high incidence of gastric mucosa and its presence in duplications even in the distal parts of the gut, (2) persistence of the embryonic relationship of the esophagus with the vertebral column is responsible for the high incidence of vertebral defects in esophageal duplications. The results of myelography in this series also support the theory of split notochord syndrome.

Child↗

Cell culture studies on human nerve sheath tumors.

The main controversy about nerve sheath tumors (NSTs) has been their histogenesis. A Schwann cell origin has been proposed by many investigators for both schwannomas and neurofibromas. However Erlandson and Woodruff observed that while schwannomas appeared to be composed predominantly of Schwann cells, neurofibromas consisted of mainly perineurial cells. In addition, variable numbers of fibroblast-like cells and intermediate cells also have been reported in the 2 lesions. Whether these represent distinct cell types or variants of Schwann cells is still debatable. In an attempt to solve this controversy, the present study was undertaken to observe the morphology and the behaviour of these tumors in culture. These studies showed that all nerve sheath tumors are basically of Schwann cell origin and that intermediate cells and fibroblasts are variants of Schwann cell. Tissue culture studies done chiefly on schwannomas showed that the morphological features of schwannomas are preserved in 'in vitro culture' condition and therefore the difference between neurofibroma and schwannoma appears to be due to inherent differentiating property of the Schwann cells along with some environmental stimulus.

Adolescent↗

Spectrum of ulcerative colitis in north India.

We analyzed retrospectively the characteristics of 87 patients with ulcerative colitis seen in a tertiary care center in north India. Ulcerative colitis was diagnosed on the basis of clinical features, sigmoidoscopy, rectal biopsy, and exclusion of microbiological causes of colitis. Severe clinical disease was seen in 53 (60%) patients that correlated with extensive colonic involvement (p < 0.001), severe changes on sigmoidoscopy (p < 0.001) and histology (p < 0.001), erythrocyte sedimentation rate (ESR) of > 30 mm for the first hour, and serum albumin of < 3 g/dl (p < 0.001). Remission of the disease was maintained on sulfasalazine in 18 (20.7%) patients, whereas 47 (54%) required steroids. Proctocolectomy with ileoanal anastomosis was done in 22 (25.3%) patients in whom there was poor response to medical treatment. Postoperative complications were seen in seven (31.8%) patients, and death occurred in four (18.1%) patients. Ulcerative colitis thus commonly presents as severe disease, with the majority of patients requiring surgery due to poor response to medical therapy. This pattern of disease as seen in an academic referral hospital in north India is virtually indistinguishable from that seen in similar centers in the West. However, in our setting, proctocolectomy and ileoanal anastomosis is cost-effective in patients unresponsive to, or unable to afford, drug treatment.

Adult↗

Differential regulation of S-adenosylmethionine synthetase isozymes by gibberellic acid in dwarf pea epicotyls.

Dwarf pea epicotyls contained a single activity peak of S-adenosylmethionine (AdoMet) synthetase (isozyme I). Gibberellic acid (GA3, 1 microM) induced two additional isozymes (II and III). Cycloheximide (20 microgram/ml) blocked the appearance of GA3-induced isozymes, suggesting that it is dependent on de novo protein synthesis. Conclusive proof was obtained by labelling the isozymes II and III with [35S]SO2-(4) in vivo. The purified 35S-labelled AdoMet synthetase isozymes (II, III) showed a single protein band that coincided with the single radioactive peak on SDS-PAGE. Molecular-sieve chromatography of the isozyme I from control dwarf pea epicotyls and three isozymes of GA3-treated epicotyls on Sepharose CL-6B showed a single activity peak with an identical molecular mass of 174 kDa for each isozyme. Analysis of purified AdoMet synthetase isozymes (I, II, III) on SDS-PAGE showed a single silver-stained protein band with a molecular mass of 87 kDa. This proved the dimeric nature of all the isozymes of AdoMet synthetase which could be physically separated by ion-exchange chromatography on DE-52. In vitro molecular hybridization of physically separated isozymes by NaCl-freeze-thaw treatment method revealed that the three isozymes (I, II, III) in GA3-treated dwarf pea epicotyls are formed through the random dimerization of two different species of enzyme subunits that differ in their net charge. Thus, the two flanking activity peaks (isozymes I, III) represent homodimers, while the middle activity peak (isozyme II) is a heterodimer. Apparently, the single isozyme I in control epicotyls is a product of one gene of AdoMet synthetase (SAM 1), while three isozymes in GA3-treated epicotyls are the product of two genes of AdoMet synthetase. We speculate that the differential regulation of AdoMet synthetase in GA3-treated epicotyls is achieved by the expression of an alternate gene of AdoMet synthetase (SAM 2).

Chromatography, Gel↗

A sequential cell kinetic study of meningioma cells in primary explant culture using bromodeoxyuridine.

The present study was undertaken to evaluate the sequential BrdU-LI at weekly intervals upto four weeks in 18 primary explant cultures of meningiomas. This revealed three distinct patterns of growth which could be arbitrarily defined as 'degenerating' (group I), 'proliferating' (group II) and 'adaptive' (group III) types. Interestingly two cases of malignant and two of recurrent meningiomas fell into the 'degenerating' group I pattern. The possible explanations for the observed relatively higher in vitro LI values compared to lower in vivo values as reported in the literature and the theoretical implications of the three distinct patterns of sequential LI values are discussed.

Adult↗

A transmission and scanning electron microscopic study of tumoral and peritumoral microblood vessels in human gliomas.

The tumor microblood vessels (MBVs) of 25 cases of gliomas of varying grades were studied and compared with those in peritumoral region using both transmission and scanning electron microscopy (TEM and SEM). The TEM study revealed numerous villous projections with pinocytotic vesicles (PCVs) and large vacuoles (LVs) concentrated mainly at the luminal aspect in tumor MBVs which increased with increasing severity of edema. The peritumoral MBVs, in addition to showing some increase in villous projections on the luminal surface, also showed increased number of PCVs and LVs concentrated at the abluminal aspect with some of them even communicating with the extravascular space. The SEM study largely corroborated the TEM findings. The sites of formation of PCVs and LVs appeared as small pits or large craters on the luminal surface of the endothelial cells of tumor MBVs. We feel that the morphological evidence of increased permeability in tumor MBVs represents their role in the development of edema and that the occurrence of reverse pinocytosis in peritumoral MBVs is a distinct possibility which may be associated with resorption of edema fluid.

Astrocytoma↗

In-vivo proliferative potential of primary human brain tumors; its correlation with histological classification and morphological features: I. Gliomas.

This study was undertaken to investigate in-vivo proliferative potential of neoplastic cells in 66 cases of gliomas of different histological types following peroperative intravenous infusion of bromodeoxyuridine (BrdU). Histological typing according to the recent modification of WHO classification did not often correlate with in-vivo cell kinetics. Among the different morphological features used for the classification, necrosis, mitosis, increased cell density and increased endothelial cell proliferation showed good correlation with tumor cell labelling index (LI) (p < 0.01-p < 0.001). A preliminary follow-up study of 36 cases for a period ranging from 9 to 36 mths suggested the possibility that higher in-vivo tumor cell LI might be associated with an early recurrence. Thus in vivo BrdU LI may supplement the histological classification of gliomas and together they may help in a better assessment of their growth rate, degree of malignancy and biological behaviour which in turn facilitate the planning of therapeutic management for individual cases.

Adolescent↗

In-vivo proliferative potential of primary human brain tumors; its correlation with histological classification and morphological features: II. Nonglial tumors.

In-vivo cell kinetics study following peroperative intravenous infusion of bromodeoxyuridine (BrdU) was done in 10 cases of primitive neuroectodermal tumors (PNET) and 44 nonglial tumors of different histological types. The histological features usually regarded as indicators of aggressive behaviour were examined in these tumors and correlated with in-vivo labelling index (LI). In the case of meningiomas, increased cellularity, pleomorphism and mitosis had no correlation with LI. Benign non-recurrent meningiomas usually showed LI < or = 1% (mean 0.6 +/- 0.3%) whereas recurrent and malignant meningioma showed higher LI (mean 2.6 +/- 0.5% and 2.8 +/- 0.4% respectively). Follow-up study suggested that meningiomas having benign histological appearance with LI > 1% might have increased chance of recurrence. In cases of PNETs among different histological features, mitosis and differentiation seemed to be related to the biological behaviour. Astrocytic differentiation was associated with lower rate of proliferation. In pituitary adenomas different hormone producing and null cell adenomas showed similar low proliferative potential (0.6 +/- 0.3%). Benign nerve sheath tumors, craniopharyngioma and choroid plexus papilloma showed low in-vivo LI of less than 1%. Thus the present study revealed the inadequacies of routine histological examination in assessing the aggressiveness of the nonglial tumors, especially meningiomas. In-vivo LI may be a good supplement to histological diagnosis as well as helping to assess the prognosis and accordingly the management of individual cases.

Adolescent↗

Morphological appearance, growth kinetics and glial fibrillary acidic protein (GFAP) expression in primary in vitro explant culture of astrocytic neoplasms.

Astrocytomas of different grades of malignancy were cultured as primary explant and their sequential growth pattern, glial fibrillary acidic protein (GFAP) expression and labelling index (LI) using bromodeoxyuridine (BrdU) were assessed and correlated with the grade of malignancy of the original tumor tissue. Low-grade astrocytomas showed patterns of growth that diverged from anaplastic astrocytomas and glioblastoma multiforme. The GFAP expression decreased with increasing time in culture in all astrocytomas irrespective of the grading. Maximum GFAP was, however, expressed in the morphologically well-differentiated stellate cells. Contrary to expectations, lower BrdU LI was observed in glioblastoma multiforme in comparison to low-grade astrocytomas, which suggests some unidentified mechanism of differentiation in vitro for astrocytomas of higher grade of malignancy. Hence, in contrast to reported literature on the prognostic value of studies on primary cultures, the present study cautions the extrapolation of the in vitro findings for astrocytomas.

Adolescent↗

Phytohormonal regulation of S-adenosylmethionine synthetase by gibberellic acid in wheat aleurones.

Gibberellic acid (GA3) brought about a 3-fold stimulation of AdoMet synthetase activity in wheat aleurones. At the qualitative level, three isozymes of AdoMet synthetase were observed by DE-52 chromatography in GA3-treated wheat aleurones. In contrast, the control wheat aleurones showed a single isozyme. Thus the phytohormone (GA3, 1 microM) induced two additional isozymes of AdoMet synthetase in wheat aleurones. The activity of all the three isozymes in GA3-treated aleurones was considerably decreased by the simultaneous presence of abscisic acid (ABA, 10 microM). Cycloheximide (20 micrograms/ml) also significantly lowered the levels of the three isozymes of AdoMet synthetase in Ga3-treated aleurones, thereby suggesting the requirement of de-novo protein synthesis for the complete induction of isozymes. However, wheat aleurones excised from embryonated wheat seeds, did not require the application of GA3 for the induction of two additional isozymes of AdoMet synthetase. Apparently, the transport of GA3 from the embryo to aleurones induced two new isozymes of AdoMet synthetase. Three isozymes of AdoMet synthetase were also observed in wheat embryos excised from germinated wheat grains, without exogenous application of GA3. The molecular weight of all the three isozymes of AdoMet synthetase in wheat system is 181,000. The molecular weight of the subunit of the enzyme is 84,000. The dimeric nature of AdoMet synthetase was established by SDS-PAGE analysis of the purified enzyme. In-vitro hybridization of two flanking isozymic peaks I and III by NaCl-freeze-thaw method resulted in the appearance of an additional middle activity peak (isozyme II). However, no additional isozymic peaks were generated when isozymic peaks I and III were individually given a freeze-thaw treatment. Thus the flanking isozymic peaks I and III represent homodimers that differed in their net charge. In contrast, the middle isozymic activity peak II, when subjected to NaCl-freeze-thaw treatments yielded two additional isozymic peaks, I and III, thereby suggesting its heterodimeric nature. We envisage that the three isozymes in GA3-treated wheat aleurone layers are formed by the random dimerization of two classes of enzyme subunits. The two enzyme subunits which differ in their net charge could be the product of two genes of AdoMet synthetase (SAM1 and SAM2). Based on this assumption, we propose that a single isozyme I in water imbibed control wheat aleurones is the product of SAM1 gene of AdoMet synthetase. The occurrence of three isozymes in GA3-treated aleurones could be ascribed to the expression of an alternate gene of AdoMet synthetase (SAM2 gene).(ABSTRACT TRUNCATED AT 400 WORDS)

Chromatography, Liquid↗

Molecular cloning and sequencing of the gene for mycocerosic acid synthase, a novel fatty acid elongating multifunctional enzyme, from Mycobacterium tuberculosis var. bovis Bacillus Calmette-Guerin.

Mycocerosyl lipids are found uniquely in the cell walls of pathogenic mycobacteria. Mycocerosic acid synthase (MAS) is a multifunctional protein which catalyzes elongation of n-fatty acyl-CoA with methylamalonyl-CoA as the elongating agent (Rainwater, D. L., and Kolattukudy, P. E. (1985) J. Biol. Chem. 260, 616-623). To understand how the various domains that catalyze the reactions involved in chain elongation are organized, mas gene from Mycobacterium tuberculosis bovis BCG was cloned. A lambda gt11 library of AluI partially digested genomic DNA from the organism was screened with an oligonucleotide probe designed from the N-terminal amino acid sequence of purified MAS. Using terminal segments of inserts from positive clones as the probe, the library was rescreened and the process was repeated. Sequencing of four overlapping clones revealed a contiguous sequence of 9699 base pair(s) (bp) of mycobacterial genome containing a 6330-bp open reading frame that could code for a protein of 2100 amino acids with a molecular mass of 225,437 daltons. The authenticity of the open reading frame as that of MAS was verified by correspondence of the amino acid sequences deduced from the gene with the directly determined amino acid sequences of the N terminus and three different internal peptide fragments. By comparing the MAS amino acid sequence with the sequences in the active site regions of known fatty acid synthases and polyketide synthases the functional domains in MAS were identified. This analysis showed that the domains were organized in the following order: beta-ketoacyl synthase, acyl transferase, dehydratase-enoyl reductase, beta-ketoreductase, acyl carrier protein; no thioesterase-like domain could be found. These results establish MAS as the first case of an elongating multifunctional enzyme composed of two identical subunits that resemble the vertebrate fatty acid synthase in size, subunit structure, and linear organization of functional domains. Southern and Western blot analyses showed absence of mas gene and encoded proteins in Mycobacterium smegmatis and Escherichia coli. This result is consistent with the report that mycocerosic acid is present only in pathogenic mycobacteria.

Acyltransferases↗

S-100 staining for the diagnosis of melanoma of the anal canal.

Thirty-two cases of anorectal melanoma are presented. These were diagnosed at the Pathology Department of the All India Institute of Medical Sciences, New Delhi, over a period of 18 years from 1970 to 1988. In 13 cases the clinical diagnosis was carcinoma melanoma was suspected clinically in only 9 of the 32 cases. Epithelioid tumours constituted the majority (26/32) 81%, followed by the pleomorphic type (4/32), 12.5% and then the sarcomatoid variant (2/32) 6.2%. Staining for S-100 protein was done on 21 cases and was found positive in 16/21 or 76%.

Anus Neoplasms↗