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Biomedical subjects

S Sarkar

Publications and source records attributed to S Sarkar.

At least 181 records · Page 10Linked to original sources
PubMed1996

Photo-cross-linking of rabbit skeletal troponin I deletion mutants with troponin C and its thiol mutants: the inhibitory region enhances binding of troponin I fragments to troponin C.

Contraction of vertebrate striated muscle is regulated by the strong Ca(2+)-dependent interaction between troponin I (TnI) and troponin C (TnC). To critically evaluate this interaction, we generated four recombinant deletion fragments of rabbit fast skeletal TnI: the NH2-terminal fragment (TnI1-94), the NH2 terminus and the inhibitory region (TnI1-120), the inhibitory region and the COOH terminus (TnI96-181), and the COOH-terminal fragment (TnI122-181) containing amino acid residues 1-94, 1-120, 96-181, and 122-181, respectively. Native TnC and seven thiol mutants, containing single cysteine residues in the two globular domains and in the central helix of TnC, e.g., Cys-12, Cys-21, Cys-57, Cys-89, Cys-122, Cys-133, and Cys-158, were labeled with 4-maleimidobenzophenone, and their interaction with the recombinant TnI fragments and the synthetic inhibitory peptide (TnI98-114, residues 98-114) was studied by photo-cross-linking. Extensive cross-linking occurred between various domains of TnC and TnI. The cross-linking patterns (a) showed that both NH2- and COOH-terminal fragments of TnI are accessible to both of the globular domains of TnC, (b) indicated that linkage of the NH2- and COOH-terminal sequences to the inhibitory region of TnI (TnIir) caused marked enhancement of cross-linking with native TnC and all seven thiol mutants, and (c) identified the region in TnC where TnIir binds as that containing residues 98, 133, 158, and 57. Thus, the results suggest that TnI and TnC may adopt flexible and dynamic conformations in which multiple interactions involving various domains of the two polypeptides occur and TnIir acting as a linker facilitates these interactions. The interaction of TnI and its fragments with actin, TnC, and TnT, considered together with the biological activity indicates that residues 96-120 represent a key structural and functional region of TnI. Whereas the NH2-terminal region of TnI stabilizes binding to TnC and TnT, the COOH-terminal region stabilizes TnC and actin binding.

Amino Acid Sequence↗
PubMed1996

Negative signaling via FcgammaRIIB1 in B cells blocks phospholipase Cgamma2 tyrosine phosphorylation but not Syk or Lyn activation.

Crosslinking of the B cell antigen receptor surface immunoglobulin induces tyrosine phosphorylation and activation of the Src family and Syk tyrosine protein kinases, tyrosine phosphorylation of phospholipase Cgamma2 (PLCgamma2) and increases in intracellular second messengers inositol phosphates and Ca2+. These activation events, in conjunction with other pathways, culminate in the induction of B cell proliferation and differentiation. In contrast, co-crosslinking surface Ig with the B cell IgG Fc receptor prevents many of these activation events, including B cell proliferation and differentiation. The precise nature of the negative signal(s) derived from Fc receptors that prevent B cell activation is not known. Here, early activation events were examined in B cells stimulated via the antigen receptor alone or under co-crosslinking conditions. The data indicated a selective block in the tyrosine phosphorylation and activation of PLCgamma2 but not in activation of the upstream kinases, Syk and Lyn, under co-crosslinking conditions. We conclude that the negative signal acts directly on PLCgamma2 and is consistent with recent studies describing an activation-induced association of a phosphotyrosine phosphatase with tyrosine-phosphorylated B cell Fc receptor.

Animals↗
PubMed1996

Inflammatory and hyperproliferative skin disease in mice that express elevated levels of the IL-1 receptor (type I) on epidermal keratinocytes. Evidence that IL-1-inducible secondary cytokines produced by keratinocytes in vivo can cause skin disease.

Interleukin (IL)-1 induces a cascade of secondary cytokines in a large number of cell types in vitro, including monocytes, fibroblasts, synovial cells, and keratinocytes. Although it has been proposed that autocrine or paracrine activation of such cells by IL- 1 in situ could orchestrate a local inflammatory response, formal proof for such an hypothesis has been lacking. In an attempt to lower the threshold for secondary cytokine production in these cells in response to IL-1, we have generated transgenic mice (designated IR10) which overexpress functional type 1 IL-1 receptor in basal layer of epidermis keratinocytes. As predicted, keratinocytes from these animals were substantially more responsive to exogenous IL-1 than nontransgenic keratinocytes when stimulated in vitro. When challenged with known inducers of keratinocyte IL-1 synthesis and release, skin of IR10 mice exhibited an exaggerated inflammatory response, characterized by epidermal hyperplasia and an acute dermal inflammatory cell infiltrate. In this setting, the secondary epidermal cytokines gro-alpha and GM-CSF were strongly induced in transgenic epidermis but not in control skin. To confirm that these changes were indeed related to IL-1 mediated activation pathways, IR10 mice were crossed to a distinct line of transgenic mice that overexpress 17-kD IL-l alpha in basal keratinocytes. Double transgenic mice derived from this cross breeding experiment developed spontaneous inflammation of the skin, similar in appearance to that induced by PMA, both histologically and macroscopically, and distinct from that seen in either parental strain spontaneously. Furthermore, secondary cytokines were more strongly induced in the double transgenic than in either parental strain. These findings conclusively demonstrate the potential for functional autocrine pathways of keratinocyte activation mediated by IL-1 alpha in vivo, and suggest that level of expression of type 1 IL-1 receptor may function as a significant control point in physiologic IL-1 mediated autocrine pathways.

Animals↗
PubMed1996

Assignment of the human fast skeletal troponin T gene (TNNT3) to chromosome 11p15.5: evidence for the presence of 11pter in a monochromosome 9 somatic cell hybrid in NIGMS mapping panel 2.

Human fast skeletal troponin T (TnTf), the tropomyosin binding component of the multisubunit troponin complex, plays an important role in the Ca2+ regulation of striated muscle contraction. Specific primers designed from the 3' end of human TnTf cDNA were used to amplify an intronic region by polymerase chain reaction (PCR). This TnTf-specific PCR product was detected from two somatic cell hybrids containing human chromosomes 9 and 11, respectively, in NIGMS mapping panel 2. However, further studies with other somatic hybrid cell lines (Bios Laboratory) localized the TnTf gene (HGMW-approved symbol TNNT3) only to chromosome 11. This observation was further confirmed by fluorescence in situ hybridization with a 12-kb TnTf genomic probe generated by extended PCR, showing the sublocalization of the gene to band p15.5 on chromosome 11. This locus is of specific interest, as Beckwith-Wiedemann syndrome and various childhood and adult tumor-related abnormalities have been mapped to this region. The study also indicates the presence of an 11pter region in the NIGMS cell hybrid GM10611, which has previously been reported to contain only human chromosome 9.

Animals↗
PubMed1996

Photothermal effects on ovarian growth and function in the soft-shelled turtle Lissemys punctata punctata.

Investigations were carried out to find out the role of higher and lower ambient temperatures together with short and long photoperiods on ovarian growth and secretion of steroids (estradiol-17 beta and progesterone) in the adult female soft-shelled turtle Lissemys punctata punctata during the preparatory, recrudescent, and quiescent phases of its seasonal reproductive cycle. In each phase, females were treated with 3 degrees C higher and 3 degrees C lower than the mean highest and lowest ambient temperatures in association with short and long photoperiods for short-term (4 week) and long-term (8 weeks or more) durations. Short-term high-temperature treatment significantly stimulated ovarian growth and secretion of estradiol-17 beta during the preparatory phase. Eight-week treatment failed to exercise further stimulation over 4-week treatment, whereas 10-week treatment caused regressive changes of ovary (atresia of follicles). Both short- and long-term high-temperature treatments caused degeneration of ovarian follicles during the recrudescent phase. Low-temperature treatment slowed down ovarian growth and secretion in both the preparatory and recrudescent phases. In the quiescent phase, low- as well as high-temperature treatments exercised little influence over the refractory ovarian growth and secretion. Photoperiod appeared to have no effect on female reproduction in any phase. Thus, high temperature is suggested to have a triggering role on ovarian growth and secretion of estrogen at the early preparatory phase, but once the ovarian function sets in, high temperature seems to have a regressive rather than stimulatory effect on ovarian growth and function.

Aging↗
PubMed1996

Demonstration of the hypoglycemic action of Momordica charantia in a validated animal model of diabetes.

In an effort to establish and document the hypoglycaemic activity of Momordica charantia in validated models of diabetes, the alcoholic extract of the pulp was studied. In the normal glucose primed rat model, M. charantia fruit extract, 500 mg kg-1, depressed the plasma glucose levels by 10-15% at 1 h. Under similar conditions, tolbutamide (100 mg kg-1) caused approximately 40% reductions in plasma glucose both at 1 and 2 h. At 500 mg kg-1, the efficacy of M. charantia was 25-30% of tolbutamide. The reduction in plasma glucose in normal glucose primed rat was not accompanied by increased insulin secretion. There was no evidence of tachyphylaxis to the effect of M. charantia extract on repeated dosing. In streptozotocin diabetes rats, it improved the oral glucose tolerance causing significant (P < 0.002) reduction in plasma glucose of 26% at 3.5 h while metformin caused 40-50% reduction at 1, 2 and 3.5 h. M. charantia extract (500 mg kg-1) caused a 4-5-fold increase in the rate of glycogen synthesis from U-14C-glucose in the liver of normally fed rats. These data suggest that the mechanism of action of M. charantia could be partly attributed to increased glucose utilization in the liver rather than an insulin secretion effect. This is the first report on the effect of M. charantia in characterized and validated animal model systems known to respond to oral hypoglycaemic drugs.

Animals↗
PubMed1996

Bayesian procedures for the estimation of mutation rates from fluctuation experiments.

Bayesian procedures are developed for estimating mutation rates from fluctuation experiments. Three Bayesian point estimators are compared with four traditional ones using the results of 10,000 simulated experiments. The Bayesian estimators were found to be at least as efficient as the best of the previously known estimators. The best Bayesian estimator is one that uses (1/m2) as the prior probability density function and a quadratic loss function. The advantage of using these estimators is most pronounced when the number of fluctuation test tubes is small. Bayesian estimation allows the incorporation of prior knowledge about the estimated parameter, in which case the resulting estimators are the most efficient. It enables the straightforward construction of confidence intervals for the estimated parameter. The increase of efficiency with prior information and the narrowing of the confidence intervals with additional experimental results are investigated. The results of the simulations show that any potential inaccuracy of estimation arising from lumping together all cultures with more than n mutants (the jackpots) almost disappears at n = 70 (provided that the number of mutations in a culture is low). These methods are applied to a set of experimental data to illustrate their use.

Bacteria↗
PubMed1996

Complications of the pectoralis major myocutaneous flap in the oral cavity: a prospective evaluation of 220 cases.

A prospective study of 220 consecutive pectoralis major myocutaneous flaps used for oral cavity reconstruction from March of 1990 to February of 1991 showed that 89 patients (40.5 percent) developed flap-related complications and 33 patients (15 percent) had complications unrelated to the flap; 92 patients (42 percent) had an uneventful recovery and there were 6 (2.7 percent) postoperative deaths. Sixty patients (27 percent) developed flap necrosis, of whom only 6 (2.7 percent) had total flap loss. Major partial loss occurred in 20 patients (9 percent) and minor flap loss occurred in 34 (15.5 percent). Flap necrosis was significantly lower in the purely myocutaneous flaps (p < 0.00000) vis-à-vis the bipedicled and osteocutaneous flaps. Fistula formation, wound infection, dehiscence at the flap margin, and postoperative hematomas occurred with comparable frequency in both groups. The female gender, primary tongue cancer, subtotal or total glossectomy, bipedicling of flaps, prior chemotherapy, and presence of systemic disease (diabetes) emerged as significant risk factors for flap necrosis on multivariate analysis (p < 0.005).

Adult↗
PubMed1996

Comparative evaluation of somatic & excretory-secretory antigens of Entamoeba histolytica in serodiagnosis of human amoebiasis by ELISA.

The excretory-secretory antigens collected from the axenic culture medium and conventional somatic antigen prepared from the whole trophozoites of E. histolytica were compared for their efficacy in serodiagnosis of amoebiasis. A total of 280 sera collected from different clinically proven cases of amoebiasis and healthy subjects were analysed against both the antigens in ELISA. Both antigens showed a 100 per cent correlation in amoebic liver abscess cases, patients infected with enteropathogens other than amoeba and healthy subjects. However, excretory-secretory antigens showed slightly higher detection rate in patients suffering from acute amoebic dysentery and asymptomatic cyst passers groups. These results clearly suggested the use of excretory-secretory antigens by ELISA for serodiagnosis of amoebiasis due to its better or equal sensitivity, specificity and easier preparation compare to conventional antigen. The use of excretory-secretory antigen in serodiagnosis will not only help in performing more tests utilizing the same chemicals, but also save the cost, time and troubles for importing the foreign chemicals required for cultivation of E. histolytica.

Animals↗