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Biomedical subjects

S Patil

Publications and source records attributed to S Patil.

At least 91 records · Page 5Linked to original sources

Chimeric plant calcium/calmodulin-dependent protein kinase gene with a neural visinin-like calcium-binding domain.

Calcium, a universal second messenger, regulates diverse cellular processes in eukaryotes. Ca2+ and Ca2+/calmodulin-regulated protein phosphorylation play a pivotal role in amplifying and diversifying the action of Ca(2+)-binding domain was cloned and characterized from lily. The cDNA clone contains an open reading frame coding for a protein of 520 amino acids. The predicted structure of CCaMK contains a catalytic domain followed by two regulatory domains, a calmodulin-binding domain and a visinin-like Ca(2+)-binding domain. The amino-terminal region of CCaMK contains all 11 conserved subdomains characteristic of serine/threonine protein kinases. The calmodulin-binding region of CCaMK has high homology (79%) to alpha subunit of mammalian Ca2+/calmodulin-dependent protein kinase. The calmodulin-binding region is fused to a neural visinin-like domain that contains three Ca(2+)-binding EF-hand motifs and a biotin-binding site. The Escherichia coli-expressed protein (approximately 56 kDa) binds calmodulin in a Ca(2+)-dependent manner. Furthermore, 45Ca-binding assays revealed that CCaMK directly binds Ca2+. The CCaMK gene is preferentially expressed in developing anthers. Southern blot analysis revealed that CCaMK is encoded by a single gene. The structural features of the gene suggest that it has multiple regulatory controls and could play a unique role in Ca2+ signaling in plants.

Amino Acid Sequence↗

Role for platelet-derived growth factor-like and epidermal growth factor-like signaling pathways in gastrulation and spiculogenesis in the Lytechinus sea urchin embryo.

The mechanisms underlying sea urchin gastrulation and spiculogenesis have been sought for decades. We have identified two growth factor signaling pathways that are involved in these developmental events. Antibodies against mammalian platelet-derived growth factor (PDGF) receptor-beta inhibited gastrulation and spiculogenesis, and antibodies against human epidermal growth factor (EGF) receptor disrupted gastrulation and spicule placement in Lytechinus pictus and L. variegatus embryos. Our studies suggested that the antibodies affect development by inhibiting rather than activating the signaling pathways. Polyclonal and monoclonal antibodies against the mammalian receptors recognized specifically Lytechinus proteins of the expected size of 170-180 x 10(3) M(r). Growth factor binding assays indicated that there are approximately 1.25 x 10(4) platelet-derived growth factor-like receptors per cell at the mesenchyme blastula stage of L. pictus, and human platelet-derived growth factor bound with an apparent affinity of KD = 4.4 nM to dissociated cells at the mesenchyme blastula stage. Immunolabelling experiments showed that at the gastrula stage, the Lytechinus platelet-derived growth factor-like receptors are located on the primary mesenchyme cells, the gut, and most prominently on the secondary mesenchyme cells and the stomodeum. The epidermal growth factor-like receptors stained less intensely on the gut and primary and secondary mesenchyme cells. Both receptors are expressed on the ciliary band and the gut of the pluteus larva but only the PDGF-like receptor is expressed on the primary mesenchyme cells. Pulse studies showed that the embryos are sensitive to the platelet-derived growth factor receptor-beta and epidermal growth factor receptor antibodies from the blastula to sometime between the mesenchyme blastula and midgastrula stages. We show that antibodies enter the blastocoel as late as the gastrula stage. Our results suggest that platelet-derived growth factor-like and epidermal growth factor-like signaling pathways are involved in the early differentiation and morphogenesis of the sea urchin gut and spicules.

Animals↗

Quantification of sodium lauryl sulfate penetration into the skin and underlying tissue after topical application--pharmacological and toxicological implications.

Sodium lauryl sulfate (SLS) is known to penetrate skin and cause cutaneous irritation. Some of these effects have been well-defined using bioengineering techniques. In this study, the ability of SLS to penetrate skin was quantified in a hairless rat model. In addition, local deep tissue penetration and systemic exposure to SLS were also evaluated to assess the toxic potential of topically applied SLS. SLS was observed to penetrate directly to a depth of about 5-6 mm below the applied site. Systemic redistribution was predominantly responsible in determining concentrations of SLS in tissues deeper than 5-6 mm. Epidermal concentrations of SLS after application of 1% (34 mM) aqueous SLS solution for 24 h were above the threshold levels which are known to evoke typical skin irritation responses. Deeper underlying tissues including dermis, subcutaneous, and muscle may also be exposed to high levels of SLS. Topically applied SLS was also observed in blood and contralateral tissues but the observed levels were not likely to elicit any systemic side effects at these doses. Traces of SLS were observed in tissues 7 days after single 24 h application of SLS, which supports the prolonged barrier disruption data generated using conventional bioengineering techniques. Cumulative treatment of SLS significantly increased the concentration of this compound in the underlying epidermis. The known preferential affinity of SLS for skin lipids and proteins was further confirmed by both in vitro and in vivo results. However, in vitro studies failed to predict the underlying tissue toxicity of SLS under the patch site when compared to the in vivo results. Such quantitative pharmacokinetic-pharmacodynamic correlations may be useful predictors for effective use of surfactants as penetration enhancers in cosmetic, pharmaceutical, and industrial applications.

Administration, Topical↗

Radial spread of sodium lauryl sulfate after topical application.

PURPOSE: Since topical application of sodium lauryl sulfate (SLS) has been reported to elevate transepidermal water loss and decrease skin capacitance in areas immediately adjacent to the applied site, studies were carried out to quantify the extent of radial spread of SLS below a topically exposed site in a hairless rat model. METHODS: Fixed sites were demarcated and the levels of SLS measured around the applied site in the epidermis, dermis and the subcutaneous tissues. Underlying deep tissue penetration and radial spread of SLS in the presence and absence of a vasoconstrictor, phenylephrine, was also quantified. RESULTS: In a typical 24 hour study, the radial spread of SLS was observed to a distance of approximately 0.75 cm from the applied site. The use of phenylephrine (1:20000), did not significantly enhance either the local underlying tissue (apart from underlying epidermis) concentration or radial spread of SLS relative to no vasoconstrictor treatment. CONCLUSIONS: Given that SLS impairs barrier function of the skin, its radial spread could be explained by a passive diffusion process. Vasoconstrictor did not remarkably alter SLS penetration and radial spread possibly due to the competing effects of vasodilation (caused by SLS) and vasoconstriction (caused by phenylephrine).

Administration, Topical↗

Cholesterol mediated changes in beta-glucuronidase activities of rat theca interstitial cells and granulosa cells.

Effects of steroid hormones on beta-glucuronidase activities of granulosa cells and theca interstitial cells were studied in vitro in the presence and absence of cholesterol in minimum essential medium (MEM with Hank's salts). Conspicuous fall in the enzyme activities of both these cells were noticed during first 10 min of incubation in MEM without cholesterol and remained lower throughout the experiment. Addition of cholesterol to incubation medium maintained beta-glucuronidase activities of both the cells as observed in the cells of immature ovary immediately after isolation. 17 beta-estradiol did not affect beta-glucuronidase activities of these cells, while testosterone and progesterone suppressed the enzyme activities of these cells in the presence of cholesterol.

Animals↗

A comparison of performance of mathematical predictive methods for medical diagnosis: identifying acute cardiac ischemia among emergency department patients.

BACKGROUND: There is increasing interest in mathematical methods for the prediction of medical outcomes. Three methods have attracted particular attention: logistic regression, classification trees (such as ID3 and CART), and neural networks. To compare their relative performance, we used a large clinical database to develop and compare models using these methods. METHODS: Each modeling method was used to generate predictive instruments for acute cardiac ischemia (which includes acute myocardial infarction and unstable angina pectoris), using prospectivel-collected clinical data on 5773 patients, who presented over a two year period to six hospitals' emergency departments with chest pain or symptoms suggesting acute ischemia. This data set was then split into training (n = 3453) and test (n = 2320) sets. Of 200 available variables, modeling was restricted to those available within the first 10 minutes of emergency department care (history, physical exam, and electrocardiogram). RESULTS: When the number of variables was limited to eight, representing a practical number for input in the real-time clinical setting, the logistic regression's receiver-operating characteristic (ROC) curve area, as a measure of diagnostic performance, was 0.887; the classification tree model's ROC curve area was 0.858, and the neural network's ROC curve area was 0.902. When the number of variables used by a model was not limited, the logistic regression's ROC area was 0.905, the classification tree model's 0.861, and the neural network's 0.923. Among these models the neural networks had noticeably poorer calibration. When the outputs from each of these unrestricted models were presented to each of the other methods as an additional independent variable, the ROC areas of the new "hybrid" models were not significantly better than the original unlimited models (ROC areas 0.858 to 0.920). CONCLUSIONS: Logistic regression, classification tree, and neural network models all can provide excellent predictive performance of medical outcomes for clinical decision aids and policy models. Their ultimate limitations seem due to the availability of the information in data (a "data barrier") rather than their respective intrinsic properties. Choices between these methods would seem to be most appropriately based on the needs of the specific application, rather than on the premise that any one of these methods is intrinsically more powerful.

Adult↗

Psyllium for the reduction of cholestyramine-associated gastrointestinal symptoms in the treatment of primary hypercholesterolemia.

OBJECTIVE: To determine if the bulk-forming laxative, psyllium hydrophilic mucilloid (PHM), reduces the gastrointestinal side effects and enhances the cholesterol-lowering efficacy of cholestyramine resin in patients with primary hypercholesterolemia. DESIGN: After a dietary lead-in period and 6 weeks of treatment with cholestyramine, the study followed a double-blinded, placebo-controlled, crossover format. SETTING: Lipid clinic affiliated with a large metropolitan community hospital. PARTICIPANTS: Twenty-seven randomly selected male and female patients with a diagnosis of primary hypercholesterolemia. Entry criteria required a fasting low-density lipoprotein cholesterol (LDL-C) concentration of 4.91 mmol/L (190 mg/dL) or greater and a triglyceride concentration of less than 2.26 mmol/L. Patients using steroids, beta-blockers, thiazide diuretics, and lipid-lowering agents, or having a history of allergy to psyllium or aspartame were excluded. INTERVENTION: The study consisted of four interventional phases of 6 weeks' duration that included (1) dietary stabilization (National Cholesterol Education Program Step I Diet); (2) cholestyramine therapy (4 g twice daily); (3) cholestyramine with study medication (PHM [5.1 g twice daily] or placebo); and (4) cholestyramine with crossover to alternate study medication. MAIN RESULTS: Following the 6-week dietary lead-in phase, four patients were eliminated from the study because their fasting LDL-C concentrations fell below 4.14 mmol/L (160 mg/dL), and one patient was eliminated because testosterone therapy was initiated by his internist. The remaining 22 patients entered the cholestyramine treatment phase. Four left the study within 2 weeks because of intolerable gastrointestinal tract symptoms. The 18 patients who completed this phase demonstrated significant reductions in their plasma total cholesterol (7.27 vs 6.67 mmol/L [281 vs 258 mg/dL]) and LDL-C (5.38 vs 4.63 mmol/L [208 vs 179 mg/dL]) concentrations compared with baseline levels. The addition of PHM to the cholestyramine regimen provided a tendency toward further reductions in total cholesterol and LDL-C levels (6.67 vs 6.46 mmol/L [258 vs 250 mg/dL] and 4.63 vs 4.29 mmol/L [179 vs 166 mg/dL], respectively), although statistical significance was not achieved. Psyllium hydrophilic mucilloid significantly reduced the frequency and severity of constipation, abdominal discomfort, and heartburn. No reports of new gastrointestinal tract symptoms or untoward effects were noted with the addition of PHM. CONCLUSION: These data suggest that the addition of PHM to cholestyramine therapy may improve a patient's compliance by reducing the associated gastrointestinal tract side effects.

Abdominal Pain↗

Perceived exertion and muscle efficiency in Parkinson's disease: L-DOPA effects.

Weakness, easy fatiguing, and lack of endurance are commonly perceived by patients with Parkinson's disease (PD). Although the slowed motor repertoire in PD may underlie these experiences, other abnormalities in skeletal muscle utilization also may be involved. We investigated whether an index of metabolic efficiency during a continuous exercise task, the latency until anaerobic threshold (AT), is altered by L-DOPA (LD). While pedalling a bicycle ergometer against a uniform workload, subjects were monitored for expired O2 and CO2. As compared to an unmedicated state, LD treatment delayed AT by a mean (+/-SE) of 5.67 +/- 0.89 to 6.62 +/- 1.23 min (p < 0.05), paired t test). Subjects did not differ in their perceived exertion upon reaching AT. With relief of parkinsonism by LD, the efficiency of energy utilization is also increased in exercised skeletal muscle.

Adult↗

Effect of age and sex on the elicitation of irritant contact dermatitis.

Irritant contact dermatitis causes significant disability to numerous consumers and individuals in industry. Knowledge of the prevalence of this disease remains inadequate. Irritant contact dermatitis occurs in workers engaged in occupations where the subjects are exposed to different types and doses of irritants. The intensity of the reaction depends on the quality and the quantity of exposure. Irritant thresholds and dose responses depend on many different factors. This article reviews the clinical and physiological parameters that affect the age and sex related differences in irritant contact dermatitis.

Adolescent↗

Effect of hepatoprotective ayurvedic drugs on lysosomal enzymes during hepatic injury induced by single dose of CCl4.

Effects of single doses of kumari asav, kumari kalp, arogyavardhini and tamra bhasma on lysosomal enzymes (acid phosphatase and beta-glucuronidase) of rat liver and kidney were studied during hepatitis induced by single 0.3 ml/kg body wt dose of CCl4. Histologically all the drugs showed significant hepatoprotection. While acid phosphatase activities of liver and kidney were suppressed, activities of beta-glucuronidase were enhanced by these drugs. The results indicate that acid phosphatase and beta-glucuronidase behave differently, although they are lysosomal in nature.

Animals↗

Neural network in the clinical diagnosis of acute pulmonary embolism.

The purpose of this investigation was to test the hypothesis that computer-based pattern recognition can accurately assess the likelihood of acute pulmonary embolism (PE) based on readily obtainable clinical characteristics. Data were obtained from 1,213 patients who participated in the collaborative study of the Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED). Characteristics of the history, physical examination, electrocardiograph, chest radiograph, and arterial blood gases of patients with suspected acute PE were presented to a back propagation neural network. The 1,213 patients were divided into training set A (n = 606) and test set B (n = 607). These groups were then reversed into training set B (n = 607) and test set A (n = 606). A receiver operating characteristic (ROC) curve was constructed from PIOPED clinical assessment, and from neural network clinical assessment in groups A and B. Areas under the respective ROC curves were 0.7450, 0.7477, and 0.7324. All differences were not significant. Areas under ROC curves for PIOPED clinical assessment combined with ventilation/perfusion (V/Q) scan results were compared with neural network clinical assessment combined with V/Q scan results in groups A and B. The respective ROC areas were 0.8324, 0.8203, 0.8496 (all differences not significant). These data show that neural networks were able to predict the clinical likelihood of PE with an accuracy comparable to experienced clinicians.

Acute Disease↗

Multiple neural inductions in area opaca by grafts of the Hensen's node do not retard host chick embryo development.

Four hundred and forty four full primitive streak stage chick embryos were cultured in vitro and 261 were transplanted with 1, 3 or 5 Hensen's nodes in the area opaca. Irrespective of the number of grafts, neural induction was observed in 90% cases. The development of control and grafted embryos and the size of blastoderm area were monitored at the time of grafting and after 20 hr. We find that the induced neural tissue and differentiated tissue of graft-origin neither fuse with the host embryonic axis, nor retard its development.

Animals↗

Proficiency testing in clinical cytogenetics. A 6-year experience with photographs, fixed cells, and fresh blood.

The College of American Pathologists and the American Society of Human Genetics offer a proficiency testing program in clinical cytogenetics. Two hundred twenty-five laboratories now provide data for this survey, which was begun in 1986. Challenges have consisted of photographed metaphases, fixed lymphoblastoid cell suspensions, fresh peripheral blood, and disarranged karyotypes. The "correct" response was based on 80% or greater consensus among either the referees or the participants. Referee laboratories performed better than participants. More laboratories were able to report accurate recognition of abnormalities by using a coded list than could write the interpretation in standardized nomenclature. Deletions, unbalanced translocations, and inversions were more difficult challenges than balanced translocations or trisomies. Prenatal and lymphocyte challenges were more likely to result in consensus than were bone marrow challenges. Participants performed best on whole-blood challenges. Fixed cell suspensions were less satisfactory. Excellent quality case material is essential for a successful challenge. A grading system has been devised to separate artifacts of the survey process from proficiency variables.

Chromosome Aberrations↗

Epidermal water permeation in vertebrates: an in vitro study using tritiated water.

Structural lipids play an important role in the water holding property of the skin. beta emitters are known to pass through the skin barrier. A permeability cell was constructed and tritiated water was used to determine the rate of in vitro water permeation through the epidermis. The epidermal sheets from different classes of vertebrates were compared for water permeation. Some of the samples were treated with solvents, surfactants and conditioners, to fathom permeability changes of the epidermis after these treatments, at various time intervals. Solvent treated epidermal sheets show very high degree of permeation, both in rat pup skin and pigeon skin. Glycerol, Brij 99 and beewax treated skin shows negligible permeation; or rather, helps in retention of water compared to normal and solvent treated skin. Comparison of water permeation in rat, pigeon and lizard skin shows that the reptilian integument is more efficient, as it allows very less permeation of water compared to the avian and mammalian epidermis. These observations suggest that tritiated water could be used for determining in vitro water permeation through the epidermis.

Animals↗

Effect of hepatoprotective ayurvedic drugs on lipolytic activities during CCl4 induced acute hepatic injury in albino rats.

Daily treatment of CCl4(3 ml/kg body wt) for 7 days induced acute hepatic necrosis in albino rats. Treatment of CCl4 caused significant alterations in the activities of acid lipase, alkaline lipase, lipoprotein lipase of liver, kidney and adipose tissue and hormone sensitive lipase of adipose tissue of albino rat. Administration of hepatoprotective ayurvedic drugs (kumari asav, kumari kalp, arogyavardhini and tamra bhasma) concomitant with CCl4 counteracted the action of CCl4 on lipolytic enzymes exhibiting hepatoprotection. The possible physiological significance of alterations in lipolytic enzymes during hepatic necrosis induced by CCl4 and hepatoprotection by the above ayurvedic drugs is discussed.

Animals↗

Cysteine proteinase inhibitor in cultured human medullary thyroid carcinoma cells.

The TT cell line of human medullary thyroid carcinoma, that retains some of the differentiated functions of thyroid C cells including the synthesis and secretion of calcitonin, was found to contain and release into the culture medium cysteine proteinase inhibitor(s), cystatin(s). The major inhibitor, which is similar to, if not identical with, cystatin C, is constitutively released, or secreted, by TT cells. The rate of secretion of cystatin, quantified by titration of inhibition of papain, was stimulated by dibutyryladenosine 3':5'-cyclic monophosphate, forskolin, the calcium ionophore A 23187, and by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA). Neither forskolin nor TPA had, however, an effect on the level of the inhibitor in TT cells. Treatment with n-butyrate strongly inhibited the proliferation of TT cells, and led, in 4 to 7 days, to a doubling of the intracellular concentration of cystatins. Northern blot hybridizations to a 32P-labeled riboprobe complementary to human cystatin C cDNA indicated that cAMP, forskolin, and TPA had no effect on the steady-state levels of cystatin C mRNA. These data indicate that release of cystatin(s) from TT cells is regulated by cAMP-calcium-protein kinase C mechanisms that appear to be similar to those that regulate the secretion of calcitonin from these cells. However, in contrast to the calcitonin gene, the expression of the cystatin C gene in these cells is not regulated by cAMP or TPA. By a combination of acetone fractionation, affinity chromatography on Cm-papain-Sepharose, and gel exclusion chromatography a protein of approximately 14 kilodaltons was isolated from TT cells that reacted with antibodies against human cystatin C, and strongly inhibited papain. Cystain secreted by TT cells also had a molecular weight of 14 kilodaltons, and reacted with anti-human cystatin C antibodies. The physiologic and pathologic roles of cystatins in different cell types remain to be established. The TT cells provide a suitable cell type to study the regulation of the expression of the cystatin gene and the mechanism of cystatin release.

Blotting, Northern↗