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P Sinha

Publications and source records attributed to P Sinha.

At least 37 records · Page 2Linked to original sources
PubMed2001

Enhanced expression of human ABC-transporter tap is associated with cellular resistance to mitoxantrone.

Multidrug resistance (MDR) phenotypes have been associated with the overexpression of various members of the superfamily of ATP binding cassette (ABC) transporters. Here we demonstrate that a member of the ABC-transporter family, the heterodimer 'transporter associated with antigen processing' (TAP), physiologically involved in major histocompatibility complex class I-restricted antigen presentation, is significantly overexpressed in the human gastric carcinoma cell line EPG85-257RNOV exhibiting a mitoxantrone-resistant phenotype. This tumor cell line shows an atypical MDR phenotype in the absence of 'P-glycoprotein' or 'MDR-associated protein' overexpression but with an enforced 'breast cancer resistance protein' expression level. Transfection of both TAP subunits encoding cDNA molecules, TAP1 and TAP2, into the drug-sensitive parental gastric carcinoma cell line EPG85-257P conferred a 3.3-fold resistance to mitoxantrone but not to alternative anti-neoplastic agents. Furthermore, cell clones transfected with both, but not singularly expressed TAP1 or TAP2, reduced cellular mitoxantrone accumulation. Taken together, the data suggest that the heterodimeric TAP complex possesses characteristics of a xenobiotic transporter and that the TAP dimer contributes to the atypical MDR phenotype of human cancer cells.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2001

Platelet-activating factor (PAF)-acetylhydrolase and PAF-like compounds in the lung: effects of hyperoxia.

Platelet-activating factor (PAF)-acetylhydrolase is the enzyme modulating in tissues and biological fluids the concentration of the proinflammatory factors PAF and PAF-like oxidation products of phospholipids (PAF-like compounds). We investigated whether there is a relation between PAF-acetylhydrolase activity and the concentration of PAF-like compounds in bronchoalveolar lavage (BAL). We found that alveolar type II cells are an additional source of PAF-acetylhydrolase in BAL beside macrophages. Secretion of PAF-acetylhydrolase was stimulated by phorbol ester in alveolar type II cells but not in macrophages. Studies in BAL suggested that secreted PAF-acetylhydrolase was bound to alveolar surfactant. Exposure of rats to high oxygen concentration reduced the activity of PAF-acetylhydrolase in BAL and macrophages, but not in plasma or alveolar type II cells. In contrast, hyperoxia increased the concentration of PAF-like-compounds, lipid hydroperoxides and malonedialdehyde in plasma but not in BAL. Therefore, we conclude that neither the oxidant-induced decrease of the PAF-acetylhydrolase activity nor the direct peroxidation of surfactant lipids in the alveoli provide a likely mechanism for hyperoxia-induced lung injury. Instead, lung injury is apparently caused by lipid peroxidation in plasma rather than by high oxygen pressure in the alveoli.

1-Alkyl-2-acetylglycerophosphocholine Esterase↗
PubMed2001

Immunological dysregulation of lung cells in response to vitamin E deficiency.

Vitamin E supplementation exhibits anti-inflammatory properties. In the lung, the beneficial effects of vitamin E supplementation on inflammation and infections are well documented, but potential consequences of alimentary vitamin E deficiency to the immunological status of lung cells are not known. It is unclear if temporary vitamin E deficiency exhibits deleterious consequences or can be compensated for by other cellular antioxidants. To address this question, the alimentary vitamin E supply to rats was modified. We then investigated the effects on major histocompatibility molecule (MHC) class II, cell adhesion molecules, interleukin (IL)10, tumor necrosis factor (TNF)alpha in various lung cells. The constitutive expression of MHC class II, intercellular adhesion molecule (ICAM)-1, L-selectin, alpha5-integrin, and CD 166, was demonstrated by flow cytometry on type II pneumocytes, alveolar macrophages, and on co-isolated lymphocytes. Vitamin E depletion increased ICAM-1 and CD166 on type II cells and macrophages, whereas the expression of L-selectin increased only on macrophages. Furthermore, the vitamin E depletion increased the cellular content and secretion of IL10 in type II cells, but decreased the content and secretion of TNFalpha. Vitamin E depletion decreased the cellular vitamin E content, but did not change the activity of antioxidant enzymes (catalase, superoxide dismutase) and the glutathion (GSH)/oxidized glutathion (GSSG) ratio in alveolar type II cells. The shift of protein kinase C (PKC) from the cytosol to membranes indicates that a PKC-dependent signaling pathway may be involved in the change of the immunological status of type II cells. All these effects were reversed by vitamin E repletion. In summary, these results are clearly compatible with the view that a temporary vitamin E deficiency induces a reversible immunological dysregulation in alveolar type II cells and lung macrophages. This deficiency might predispose the lung to develop acute or chronic inflammation.

Animals↗
PubMed2001

Receiver-operating characteristic as a tool for evaluating the diagnostic performance of prostate-specific antigen and its molecular forms--What has to be considered?

BACKGROUND: Receiver-operating characteristic (ROC) analysis is often applied as evaluation tool to compare the diagnostic validity of laboratory tests. The aim of this study was to draw attention to preconditions which should be taken into account when ROC analysis is used to assess the diagnostic performance of total prostate-specific antigen (tPSA) and its molecular forms in differential diagnosis between prostate cancer and benign prostatic hyperplasia (BPH). METHODS: Using a standard software (GraphROC for Windows), ROC analyses were performed and the areas under the curves were calculated for four hypothetical pairs of groups. Every group included 40 patients with prostate cancer and with BPH showing different tPSA concentrations (range of 2-10 microg, but similar free-to-total PSA ratios (fPSA%). RESULTS: The area under the fPSA% ROC curve showed the highest value, whereas the areas under the tPSA ROC curves were dependent on the distributions of tPSA concentrations in the patients. The ability of fPSA% to improve the differential diagnosis between prostate cancer and BPH in comparison to tPSA was not furthermore evident, if the prostate cancer group included more patients with higher tPSA concentrations than the BPH group. CONCLUSIONS: When the diagnostic performance of tPSA and its derivatives like molecular forms in patients with prostate cancer and BPH should be compared by ROC analysis, a matching procedure is recommended prior to ROC analysis to compensate the effect of possible unequal tPSA distributions in both groups. Each BPH (or PCa) patient should be matched with a PCa (or BPH) patient with nearest tPSA concentration so that an optimum of overlapping tPSA concentrations in both groups can be achieved.

Diagnosis, Differential↗
PubMed2001

An economised craniofacial identification system.

It has been attempted to develop an economised craniofacial identification system, as a special automated version of photo/video superimposition technique, that can deal with common cases of personal identification with the aid of a skull and a nearly front view face photograph of the suspected victim. The proposed method is economic in respect of (i) cost of hardware configuration, (ii) processing time as well as (iii) manual labour involved. Over and above, it has got a capability to take care of ambiguities due to soft tissue thickness during the selection of facial features, which is a part of the procedure. In order to reconstruct a 2-D cranial image, superimposable over the facial one, the new method does not need any reconstruction of a digitised 3-D cranial image. It works simply by a suitable segment-wise processing of a 2-D cranial image with the aid of the symmetry perceiving adaptive neuronet (SPAN), that has recently been introduced in connection with nearly front view facial image recognition. The final comparison of the facial and the superimposable cranial images is as versatile as the same for facial image recognition by SPAN.A practical application of this extended version of SPAN has been demonstrated in the present paper.

Child↗
PubMed2001

Characterization of the differential protein expression associated with thermoresistance in human gastric carcinoma cell lines.

Resistance to chemotherapeutic agents is one of the major problems faced during palliative therapy of tumor cells. Thus, chemotherapy is frequently combined with other modes of therapy such as radiation therapy and/or hyperthermia. Tumor cells respond to heat stress with development of thermotolerance and the interactions between chemo- and thermoresistance phenomena are not clearly understood. In this paper, we analyze the differential protein expression in vitro in human stomach cancer cells, their chemoresistant and thermoresistant counterparts using proteomics. The immediate aim was to identify sets of proteins that may lead to the development of thermoresistance. Based on these results, we aim to develop functional tests and methods for the modulation of thermoresistance and chemoresistance phenomena that may assist the therapy of inoperable cancers.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2001

A new silver staining apparatus and procedure for matrix-assisted laser desorption/ionization-time of flight analysis of proteins after two-dimensional electrophoresis.

We report on a new silver stain especially developed for staining large gels (25 cm x 20 cm) from the Hoefer ISO-DALT system for matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis of proteins. The staining protocol can be summarized as follows: the gels are sensitised in tetrathionate/potassium acetate solution and washed several times in distilled water. After impregnation with silver nitrate, the silver is reduced in the presence of potassium carbonate, thiosulphate and formaldehyde. The staining procedure is stopped with Tris/acetate after which the gels are rinsed and stored in water before spot picking for MALDI-TOF analysis is performed. This protocol has several advantages over existing ones. The gels are stained in a new apparatus that reduces gel handling to a minimum thus also reducing the contamination with keratins to a minimum. The development times in potassium carbonate are very long (up to 40 min) thus improving batch-to-batch reproducibility. Only the surface of the proteins is stained and the silver can be oxidized, thereafter MALDI-TOF can be performed with protein loads as little as 100 micrograms per gel.

Cell Line↗
PubMed2001

Proteomics for studying cancer cells and the development of chemoresistance.

Extensive studies during the last decades have identified several mechanisms through which cells escape the cytotoxic effects of a variety of chemotherapeutic drugs. One type of drug resistance is called multidrug resistance (MDR), because selection with one anticancer drug leads to cross-resistance with a wide range of other drugs. These MDR cells express frequently plasma transport proteins like p-glycoprotein. But cellular resistance to chemotherapy is multifactorial and may be affected by the cell cycle stage and proliferation status, biochemical mechanisms such as detoxification, cellular drug transport, or DNA replication and repair mechanisms. Several laboratory techniques, such as polymerase chain reaction, immunocytochemistry, flow cytometry, blotting, and fluorescent microscopy have been used for the identification of MDR markers and mechanisms. We review the possibilities in studying cancer biology and development of chemoresistance in cancer treatment using the proteomic approach.

Animals↗
PubMed2001

The IML3/MCM19 gene of Saccharomyces cerevisiae is required for a kinetochore-related process during chromosome segregation.

The mcm19 mutation in budding yeast affects minichromosome maintenance. In this work we have shown that this mutation leads to defects in the segregation of minichromosomes and chromosomes. The mutant cells show defective kinetochore function as judged by three criteria-- relaxation of the transcriptional block normally associated with a CEN box, stable maintenance of a dicentric plasmid in mutant cells, and mild sensitivity to the antimicrotubule drug benomyl. The MCM19 gene has been cloned and found to be the same as IML3, which codes for the ORF YBR107C. Deletion of the gene was not lethal, nor did it confer any growth defects on the mutant cells. However, the mcm19 null mutation conferred growth defects in the presence of a mutation in the TUB1 gene coding for alpha-tubulin. Two-hybrid experiments showed an interaction between Im13p/Mcm19p and the kinetochore protein Ch14, indicating that the Im13/Mcm19 protein has a role in kinetochore function.

Benomyl↗
PubMed2001

Semiquantitative analysis of types I and III collagen from tendons and ligaments in a rabbit model.

The present study presents a simple and reliable micro-method for the semiquantitative analysis of types I and III collagen in tendons and ligaments in a rabbit model. After pretreatment of the analytical material by homogenization, a double cyanogen bromide cleavage was performed and the peptide fragments were visualized by polyacrylamide gel electrophoresis and silvcr staining. On the basis of this procedure, the method presented here can be used to analyze very small amounts of sample material (less than 10microg) by electrophoresis. The results of the study showed that type I collagen is predominant in the ligaments and the tendons of the knee, e.g., medial and lateral collateral ligaments. anterior and posterior cruciate ligaments. patellar tendon, achilles tendon, and semitendinosus tendon. However, a markedly higher proportion of type III collagen was detected in the ligaments (approximately 10%) than in the tendons (approximately 5%). The ligaments differ markedly from the tendons in biochemical mapping; the ligaments are functionally and metabolically the more active tissue and have a higher adaptation potential.

Animals↗
PubMed2001

Long-term outcomes after cardiac transplantation: an experience based on different eras of immunosuppressive therapy.

BACKGROUND: Constantly changing practices in heart transplantation have improved posttransplant survival in patients with end-stage heart disease. The objective of this study was to evaluate long-term outcomes in different eras of immunosuppressive therapy after cardiac transplantation at a single center during a two-decade period. METHODS: A retrospective review of 1,086 consecutive cardiac allograft recipients who underwent transplantation between 1977 to 1999 was performed. Patients were divided into four eras based on type of immunosuppressive therapy: era 1 = steroids, azathioprine (n = 26, February 1977 to March 1983), era II = steroids, cyclosporine (n = 43, April 1983 to April 1985), era III = cyclosporine, steroids, azathioprine (n = 752, April 1985 to December 1995), era IV = cyclosporine, steroids, mycophenolate mofetil (n = 315, January 1996 to October 1999). RESULTS: The actuarial survival of the entire cohort of 1,086 patients undergoing cardiac transplantation was 79%, 66%, and 49% at 1, 5, and 10 years, respectively. There were significant trends in recipient age and gender distribution among the four eras with increasing proportion of older age (> 60 years) and female recipients in eras III and IV (p = 0.001 and 0.02). Early mortality and long-term survival improved significantly over all eras (p < 0.001). Rejection as a cause of death decreased over time (era I, 24%; era II, 21%; era III, 15%; era IV, 9%; p = 0.02), whereas the contribution of transplant coronary artery disease as a cause of death remained unchanged. CONCLUSIONS: Cardiac transplantation provides satisfactory long-term survival for patients with end-stage heart failure. The improving outcomes in survival correlate with improved immunosuppressive therapy in each era. Although the reasons for improvement in survival over time are multifactorial, we believe that changes in immunosuppressive therapy have had a major impact on survival as evidenced by the decreasing number of deaths due to rejection.

Actuarial Analysis↗
PubMed2001

Systematic instrumental errors between oxygen saturation analysers in fetal blood during deep hypoxemia.

BACKGROUND: During a study of artificially produced deep hypoxemia in fetal cord blood, systematic errors of three different oxygen saturation analysers were evaluated against a reference CO oximeter. METHODS: The oxygen tensions (PO2) of 83 pre-heparinized fetal blood samples from umbilical veins were reduced by tonometry to 1.3 kPa (10 mm Hg) and 2.7 kPa (20 mm Hg). The oxygen saturation (SO2) was determined (n=1328) on a reference CO oximeter (ABL625, Radiometer Copenhagen) and on three tested instruments (two CO oximeters: Chiron865, Bayer Diagnostics; ABL700, Radiometer Copenhagen, and a portable blood gas analyser, i-STAT, Abbott). The CO oximeters measure the oxyhemoglobin and the reduced hemoglobin fractions by absorption spectrophotometry. The i-STAT system calculates the oxygen saturation from the measured pH, PO2, and PCO2. The measurements were performed in duplicate. Statistical evaluation focused on the differences between duplicate measurements and on systematic instrumental errors in oxygen saturation analysis compared to the reference CO oximeter. RESULTS: After tonometry, the median saturation dropped to 32.9% at a PO2=2.7 kPa (20 mm Hg), defined as saturation range 1, and to 10% SO2 at a PO2=1.3 kPa (10 mm Hg), defined as range 2. With decreasing SO2, all devices showed an increased difference between duplicate measurements. ABL625 and ABL700 showed the closest agreement between instruments (0.25% SO2 bias at saturation range 1 and -0.33% SO2 bias at saturation range 2). Chiron865 indicated higher saturation values than ABL 625 (3.07% SO2 bias at saturation range 1 and 2.28% SO2 bias at saturation range 2). Calculated saturation values (i-STAT) were more than 30% lower than the measured values of ABL625. CONCLUSION: The disagreement among CO oximeters was small but increasing under deep hypoxemia. Calculation found unacceptably low saturation.

Blood Gas Analysis↗
PubMed2001

Role of motion integration in contour perception.

Contours are believed to play a key role in the visual analysis of scenes by the primate brain. In dynamic scenes, the presence of contours is often signaled by discontinuities in motion fields. However, it is unclear whether the motion fields over which the visual system extracts discontinuities, correspond to the local optic-flow or the pattern motion fields obtained by integrating local estimates. A resolution of this issue would provide important clues about the organization of visual motion and form analysis processes. In this paper, we present experimental evidence which suggests that the perception of motion defined contours is strongly dependent on motion integration - an operation that is believed to take place relatively late in the visual stream.

Form Perception↗
PubMed2001

Child hospitalization: an opportunity for parental smoking intervention.

BACKGROUND: Hospitalization with tobacco-related illness increases smokers' interest in cessation. Because parental smoking increases the child's risk of developing respiratory and other illnesses, a child's hospitalization might motivate a smoking parent to consider changing smoking behavior. It is unclear if parents would be receptive to smoking-cessation interventions at the time when their child is hospitalized. METHODS: In March 1999, parents of 298 consecutive children admitted to the medical services at Children's Hospital Boston were interviewed to determine the smoking status of household members. Smoking parents were invited to complete a 35-item questionnaire regarding personal smoking history and acceptability of three types of cessation interventions. RESULTS: Sixty-five smoking parents were identified among the 298 admissions; 62 of 65 (95%) participated in the survey. Among respondents, only 15% had ever participated in any smoking-cessation program, and only 31% had ever used a medication to try to quit. Although 78% of parents were willing to speak with a counselor about their smoking while their child was in the hospital, and 74% would enroll in a telephone-based smoking-cessation program, only 26% were interested in a free program requiring travel back to the hospital. All parents believed that pediatricians should offer parental smokers the chance to participate in a smoking-cessation program. CONCLUSIONS: At the time of a child's hospitalization, parents are willing to enroll in smoking interventions that include in-hospital and telephone counseling but not to travel back to the hospital. A child's hospitalization may provide a unique opportunity to enroll parents who smoke into cessation programs.

Adult↗
PubMed2001

Spatial regulation of DELTA expression mediates NOTCH signalling for segmentation of Drosophila legs.

The Notch (N) signalling pathway is recruited for segregation of cell fates in a number of Drosophila tissue types. We show here that N dependent segmentation of Drosophila legs is regulated by a dynamic pattern of expression of its ligand, DELTA (DL). During third larval instar and early stages of pupation, high levels of DL expression is seen in stripes of cells in the leg imaginal discs which later form the proximal borders of leg joints. These domains also displayed heightened Dl enhancer activity. During subsequent stages of pupation, following segmentation of the leg primordium, DL expression becomes uniform throughout these segments barring the joints. We further show that regulatory Dl mutations or mis-expression of DL abolish leg segmentation. Domains of N signalling for segmentation of legs of flies are thus set up by a stringent spatial regulation of expression of its ligand at the segment border. Further, a comparable role of DL in antennal development reveals a common paradigm of DL-N signalling for segmentation of appendages in flies.

Animals↗
PubMed2001

The management of eclampsia by obstetric anaesthetists in UK: a postal survey.

Eclampsia and pre-eclampsia are major causes of morbidity and mortality in the obstetric population. The latest triennial report on Confidential Enquiries into Maternal Deaths found hypertensive disorders of pregnancy to be the second most common cause of maternal deaths directly attributable to pregnancy. The management of eclampsia includes the control and prevention of further convulsions by pharmacological methods but the choice of drugs may vary. Current evidence supports the use of magnesium sulphate as the drug of first choice for treating eclampsia. Until recently, UK clinicians used other anticonvulsants for this purpose. Questionnaire studies conducted amongst UK obstetricians published in 1991 and 1998 showed that magnesium sulphate was used in the management of severe pre-eclampsia by 2% and 40% of clinicians respectively. We conducted a survey among lead obstetric anaesthetists in 264 obstetric units in the UK to examine their current practice. We also asked how magnesium was used and included a question on the use of nimodipine. The response rate was 86%. Our results show that magnesium sulphate is used for the treatment of eclampsia in 90% of units and for severe pre-eclampsia in 68%. Most administered magnesium for 24-48 h while nimodipine was used by very few units.

Journal Article↗
PubMed2001

Gallbladder stone shown on a hepatobiliary scan.

Hepatobiliary scans are performed frequently to identify gallbladder-related disease. Although gallbladder stones are a common cause of hepatobiliary disease, they are visualized directly only rarely on hepatobiliary scans. Large gallbladder stones produced a photopenic defect in the gallbladder in a hepatobiliary scan.

Biliary Tract↗