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Biomedical subjects

H Das

Publications and source records attributed to H Das.

At least 19 recordsLinked to original sources

Epidemiology of chronic dacryocystitis and success rate of external dacryocystorhinostomy in Nepal.

PURPOSE: To study the epidemiology of chronic dacryocystitis and the success rate of external dacryocystorhinostomy (DCR) in Nepal. PATIENTS AND METHODS: This is a retrospective descriptive and interventional case series. A total of 662 records of patients with chronic dacryocystitis due to nasolacrimal duct obstruction, who were treated by means of external DCR without bicanalicular silastic tube intubation, were studied. STATISTICS: Mean for age with standard deviation and confidence intervals were calculated. The significance of the difference in means was calculated using the unpaired t-test. RESULTS: Of 662 patients, 67.6% were female. The mean age of the patients was 27.4 +/- 13.7 years (SE = 0.53, 95% CI = 26.34-28.46). Chronic dacryocystitis due to nasolacrimal duct obstruction was found in 86 patients (13.0%) from the high hills (mean age = 35.3 +/- 11.4 years, 95% CI = 32.85-37.75), but most patients (576, 87.0%) were from the subtropical lowlands with a monsoon climate (mean age = 26.9 +/- 13.3 years, 95% CI = 25.8-28.0; p = < 0.01). An overall success rate of 88.6% (89.8% for high-hill patients and 87.4% for those from the lowlands) was obtained after DCR. CONCLUSION: Chronic dacryocystitis is more common in the subtropical plains of Nepal with a monsoon climate than in the high hills. It is predominantly found in pre-menopausal females. External DCR without bicanalicular silastic tube intubation is an effective method for treating chronic dacryocystitis due to NLDO in these patients.

Adolescent↗

Human V gamma 2V delta 2 T cells produce IFN-gamma and TNF-alpha with an on/off/on cycling pattern in response to live bacterial products.

Whereas cytokine production in alphabeta T cells is rapidly regulated by exposure to peptide Ag, the mechanisms regulating cytokine production by gammadelta T cells are unknown. In this study, we demonstrate that human Vgamma2Vdelta2 T cells produce IFN-gamma and TNF-alpha as early as 2 h after Ag exposure, and that they produce these cytokines in a dose- and time- dependent manner in response to stimulation with a live bacterial product, iso-butylamine (IBA), but not to dead bacteria or LPS. gammadelta T cells began, ceased, and then resumed IFN-gamma and TNF-alpha generation in an on/off/on cycling pattern, both in vitro and in vivo, depending on the presence or absence of IBA. IFN-gamma and TNF-alpha, whose optimum production was dependent on IBA-stimulated gammadelta T cells, were critical for monocyte-mediated killing of Escherichia coli. By limiting cytokine production to periods of direct contact with live bacteria, gammadelta T cells focus their resources at the site of infection, while limiting systemic immunopathology. Thus, human gammadelta T cells may mediate innate resistance to extracellular bacteria via tightly regulated cytokine production without necessarily expanding in number.

Animals↗

Vgamma2Vdelta2 T-cell receptor-mediated recognition of aminobisphosphonates.

Aminobisphosphonates, potent derivatives of bisphosphonates, are frequently used for the treatment of conditions such as osteoporosis and bone metastases that are characterized by excessive osteoclastic bone resorption. Using T-cell receptor (TCR) transfer studies, we show that recognition of antigenic aminobisphosphonates that are known to stimulate human gammadelta T cells in vitro and in vivo (potency: risedronate > alendronate > pamidronate) requires expression of the Vgamma2Vdelta2 TCR and is thus Vgamma2Vdelta2 TCR-dependent. Myeloma cells or monocytes pulsed with risedronate and then washed rendered these target cells sensitive to lysis by a Vgamma2Vdelta2 T-cell clone or cell line. These results suggest that Vgamma2Vdelta2 TCR-dependent recognition leading to direct cytolysis of aminobisphosphonate-sensitized osteoclast or tumor targets may be a mechanism whereby aminobisphosphonate treatment of cancers metastatic to bone decreases osteoclastic activity and tumor burden and also may account for the decreased osteoclastic activity associated with successful treatment of osteoporosis.

Alendronate↗

MICA engagement by human Vgamma2Vdelta2 T cells enhances their antigen-dependent effector function.

Vgamma2Vdelta2 T cells comprise 2%-5% of human peripheral blood T cells, recognize ubiquitous nonpeptide antigens, and expand up to 50-fold during microbial infection. It is not clear why these Vgamma2Vdelta2 T cells expand only after microbial infection. We show here that the stress-inducible molecule, MICA, is induced on the surface of dendritic and epithelial cells by infection with M. tuberculosis in vitro and in vivo. MICA engagement by the activating receptor, NKG2D, present on Vgamma2Vdelta2 T cells, resulted in a substantial enhancement of the TCR-dependent Vgamma2Vdelta2 T cell response to nonpeptide antigens and protein superantigens alike. Thus, a MICA-NKG2D interaction may be necessary for an effective innate immune response to microbe-associated antigens that also are constitutively present in vivo.

Cytokines↗

Kinetic analysis of cytokine gene expression in patients with GVHD after donor lymphocyte infusion.

Patients who receive a donor lymphocyte infusion (DLI) for the treatment of relapsed leukemia after allogeneic BMT (alloBMT) often developed GVHD. To determine whether cytokines might have a role in GVHD, an intensive kinetic analysis of in vivo cytokine gene expression was performed on PBMC from three such patients. Expression of IL-1beta, IL-2, IFN-gamma, IL-4, IL-5, IL-8, IL-10, IL-12, TNF-alpha, and IL-2Ralpha was examined using a sensitive semi-quantitative reverse transcription (RT)-PCR assay system. Six normal controls were also analyzed for comparison. Expression of type 1 T helper (Th1) cytokines, IL-2 and IFN-gamma was greatly increased in all three patients. In particular, the changes in IL-2 gene expression correlated well with disease progression, suggesting that IL-2 has a critical role in the development of GVHD. Although the pattern of type 2 T helper (Th2) cytokine gene expression differed in each patient, the expression of IL-4 was inversely related to expression of Th1 cytokines. These results suggest that Th1 dominates in the development of human clinical GVHD.

Adult↗

Robot-assisted microsurgery: a feasibility study in the rat.

OBJECTIVE: Telerobotic surgery is a novel technology that can improve a surgeon's manual dexterity as well as the results achieved with microsurgical procedures. METHODS: A prototype Robot-Assisted MicroSurgery (RAMS) microdexterity enhancement system developed by the Jet Propulsion Laboratory and MicroDexterity Systems, Inc., was tested in 10 rats. Carotid arteriotomies were created and closed using either the RAMS system or conventional microsurgical techniques. The time required, the technical quality (vessel patency and suture line integrity), the error rate, and subjective difficulty were compared. RESULTS: All procedures were successfully completed using the RAMS system to manipulate the vessel but not to hold the needle or place the sutures. The precision, technical quality, and error rate of telerobotic surgery were similar to those of conventional techniques. However, the use of the RAMS system was associated with a twofold increase in the length of the procedure. CONCLUSION: Surgery using a microdexterity enhancement system, or RAMS prototype, is feasible. With further development, such as a stereotelevisualization and haptic feedback system, this system could be used for telerobotic surgery in neurosurgical practice.

Animals↗

Antibacterial effect of human V gamma 2V delta 2 T cells in vivo.

V gamma 2V delta 2 cells, a class of T cells found only in primates, are reactive to nonpeptide organophosphate and alkylamine antigens secreted by bacteria and parasites. These cells make up 2-5% percent of human peripheral blood T cells but expand to make up 8-60% of peripheral blood T cells during bacterial and parasitic infections. We show here, using a chimeric severe combined immunodeficiency (SCID) mouse (hu-SCID) model, that human V gamma 2V delta 2 T cells mediate resistance to extracellular gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli and Morganella morganii) bacteria, as assessed by survival, body weight, bacterial loads, and histopathology. Surprisingly, this bacterial resistance was evident 1 day after infection, and bacteria were cleared well before gamma delta T cell expansion was detected 6 days after infection. Decreased resistance in V delta 2 T cell-depleted hu-SCID mice correlated with decreased serum IFN-gamma titers. Intravenous treatment of infected, reconstituted hu-SCID mice with pamidronate, a human V gamma 2V delta 2 T cell-specific aminobisphosphonate antigen, markedly increased the in vivo antibacterial effect of V gamma 2V delta 2 T cells. Therefore, this large pool of antigen-specific, yet immediately reactive memory human V gamma 2V delta 2 T cells is likely to be an important mediator of resistance against extracellular bacterial infection and may bridge the gap between innate and acquired immunity.

Animals↗

Quantitation of minimal residual disease in t(8;21)-positive acute myelogenous leukemia patients using real-time quantitative RT-PCR.

t(8;21) is one of the common chromosomal translocations in acute myelogenous leukemia (AML). Using a recently developed real-time quantitative polymerase chain reaction (PCR) system, we analyzed the minimal residual disease (MRD) in bone marrow samples from seven AML patients with t(8;21) at different time points during the clinical courses of their disease. Four of these patients received chemotherapy and allogenic bone marrow transplantation (allo-BMT), and the other three were treated with chemotherapy alone. Two of the patients that received allo-BMT suffered a relapse. In these patients, the levels of AML1-MTG8 mRNA expression were shown to quantitatively increase. After re-induction chemotherapy and donor lymphocyte infusion therapy, AML went into remission and the expression levels decreased. In the other two patients receiving allo-BMT, the disease went into remission and the level of AML1-MTG8 mRNA expression remained under the detectable range. The other three patients received several courses of chemotherapy, without allo-BMT, and all of them clinically reached the hematological and cytogenetic remission state. However, there were low but detectable levels of MRD in their bone marrow samples. These results suggest that the real-time quantitative PCR assay is very useful for the monitoring of MRD and detecting an early relapse. This assay may also be useful in determining the quantitative difference in myelo-ablative activity between the chemotherapy alone and chemotherapy in conjunction with allo-BMT.

Adult↗

Trophoblastic cells expressing human chorionic gonadotropin genes in peripheral blood of patients with trophoblastic disease.

We attempted to identify the cells expressing alpha and beta subunits of human chorionic gonadotropin (hCG) in the peripheral blood of patients with trophoblastic disease and normal pregnant women by using reverse transcriptase polymerase chain reaction (RT-PCR) and Southern blot. By this method, the mRNAs of hCG alpha and hCG beta were detected in the peripheral blood mononulear cells (PBMNC) from 3 of 7 hydatidiform mole (mole) and 1 of 4 choriocarcinoma patients as well as from normal pregnant women during the first trimester. None of the mRNAs of hCG subunits was detected in the PBMNC from healthy male and nonpregnant healthy women examined. The expression of hCG alpha and hCG beta in patients with trophoblastic disease and normal pregnant women almost correlated with their plasma levels of intact hCG. The present study indicates that the cells expressing hCG alpha and hCG beta, which virtually represent trophoblasts, are circulating in the peripheral blood of patients with trophoblastic disease as well as of normal pregnant women.

Blotting, Southern↗

Quantitation of Fas and Fas ligand gene expression in human ovarian, cervical and endometrial carcinomas using real-time quantitative RT-PCR.

Alterations in the expression of Fas (CD95/APO-1) and its ligand (FasL) have been demonstrated in various types of cancers as a mechanism for tumour cell to escape from the immune system. In the present study, we evaluated the expression of the Fas and FasL genes in a wide range of primary gynaecological carcinomas. These included 31 ovarian, 29 cervical and 25 endometrial carcinoma tissues as well as four ovarian and three cervical carcinoma cell lines. Our real-time quantitative reverse transcription polymerase chain reaction analysis revealed that down-regulation of Fas expression is more prominent than the up-regulation of FasL expression in all types of gynaecological cancer studied. This down-regulation of Fas expression was also true for the seven carcinoma cell lines. Only one cervical carcinoma cell line, DoT, exhibited a high level of FasL expression. These results indicated that down-regulation of Fas expression is a common abnormality in many types of cancers including gynaecological cancers, whereas an increase in FasL expression is not a common phenomenon in these cancers.

Cervix Uteri↗

Kinetics of serum cytokines after allogeneic bone marrow transplantation: interleukin-5 as a potential marker of acute graft-versus-host disease.

Cytokine levels in sera from 14 patients undergoing allogeneic bone marrow transplantation (alloBMT) or donor lymphocyte infusion (DLI) were sequentially measured to evaluate the roles of cytokines in clinical graft-versus-host disease (GVHD). In clinical courses, interleukin (IL)-1 alpha, IL-1 beta, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma were measured by enzyme-linked immunosorbent assay. Among the evaluable cases, 6 patients developed acute GVHD. Serum IL-5 levels increased to more than 100 pg/mL in 5 of the 6 patients before, or simultaneously with, the clinical manifestation of acute GVHD. Elevation of IL-5 was transient in 3 patients. In the other 2 patients who showed regimen-related toxicity and/or thrombotic microangiopathy as well as acute GVHD, remarkable and sustained elevation of the serum IL-5 level was observed. In 7 patients without acute GVHD, IL-5 levels remained below 100 pg/mL. An association of acute GVHD was less prominent with TNF-alpha than with IL-5 in our study. Elevation of IL-6 was associated with infections. In 2 patients with severe extensive chronic GVHD, serum IL-10 was elevated in parallel with exacerbation of clinical symptoms. Our findings suggest that an elevated serum IL-5 level may be a marker of acute GVHD.

Acute Disease↗

Soluble fas ligand in natural killer cell lymphoma.

We measured serum soluble Fas ligand (sFasL) in a patient with natural killer cell lymphoma, and investigated relationship between sFasL and liver dysfunction. An elevated level of sFasL was decreased after local radiation therapy, and liver function improved. When lymphoma relapsed, liver dysfunction reappeared and the level of sFasL increased parallelly. Lymphoma cells expressed mRNA of FasL. This suggested that this liver dysfunction was induced by some remote effectors, and sFasL was one of candidates of these effectors.

Adult↗

Evaluation of a telerobotic system to assist surgeons in microsurgery.

A tool was developed that assists surgeons in manipulating surgical instruments more precisely than is possible manually. The tool is a telemanipulator that scales down the surgeon's hand motion and filters tremor in the motion. The signals measured from the surgeon's hand are transformed and used to drive a six-degrees-of-freedom robot to position the surgical instrument mounted on its tip. A pilot study comparing the performance of the telemanipulator system against manual instrument positioning was conducted at the University of Southern California School of Medicine. The results show that a telerobotic tool can improve the performance of a microsurgeon by increasing the precision with which he can position surgical instruments, but this is achieved at the cost of increased time in performing the task. We believe that this technology will extend the capabilities of microsurgeons and allow more surgeons to perform highly skilled procedures currently performed only by the best surgeons. It will also enable performance of new surgical procedures that are beyond the capabilities of even the most skilled surgeons.

Evaluation Studies as Topic↗

Carbohydrate analysis of bradyrhizobial (NC92) lipopolysaccharides by high performance-anion exchange chromatography with pulsed amperometric detection.

Composition analysis of monosaccharides of Sepharose 4B purified NC 92 LPS and the polysaccharides fractions from Sephadex G-50 chromatography was performed by high performance anion exchange chromatography using pulsed amperometric detection. Rhamnose, mannose, galactose and glucose are present in a substantial amount in the purified LPS (Pk I). High molecular weight purified polysaccharides (PS I) obtained after sephadex gel filtration of the purified LPS (Pk I) acid hydrolysate showed an increase in glucose:galactose ratio. This indicates the presence of the peanut root lectin (PRA II) specific sugar in higher proportion on the O-antigen part of the LPS molecule, which may aid in the critical recognition reaction.

Bradyrhizobium↗

Levels of soluble FasL and FasL gene expression during the development of graft-versus-host disease in DLT-treated patients.

Three patients with different clinical symptoms of graft-versus-host disease (GVHD) who had received donor lymphocyte transfusion (DLT) for the treatment of relapsed leukaemia after an allogeneic bone marrow transplantation (BMT) from HLA-matched sibling donors were analysed for the presence of soluble FasL (sFasL) in the sera and for the expression of the Fas ligand (FasL) gene in the peripheral blood mononuclear cells (PBMNC). Two patients who demonstrated liver damage with increased levels of serum bilirubin showed significantly increased levels of serum sFasL. The increase in the sFasL level was observed prior to the increase in the bilirubin during the clinical courses of both patients. The high dose of methyl predonisolone administered to one of these patients greatly reduced the levels of sFasL in the serum. The bilirubin levels were also reduced thereafter. The third patient (without liver damage) did not show any increase in the serum sFasL level. The expression of the FasL gene in the PBMNC of these three patients was examined. All three patients showed increased levels of the FasL gene expression during their clinical courses. However, only one patient showed a parallel alteration of FasL gene expression with sFasL in the serum. These cases provide evidence that the Fas/FasL system is closely associated with human GVHD, especially in the development of liver GVHD.

Adult↗

Evaluation of the anticancer property of a new alpha-methylene-gamma-lactone derivative of phthalimide.

The anticancer property of a new alpha-methylene-gamma-lactone derivative of phthalimide (2, NSC 640168) was evaluated in two murine ascitic tumors namely Ehrlich ascites carcinoma (EAC) and sarcoma-180 (S-180) by in vivo screenings and in a battery of human tumor cell lines by in vitro screening. It was found that the compound has exhibited marginal to moderate in vivo activity in EAC and S-180, respectively, and significant in vitro cytotoxicity in SF-268, a human CNS tumor cell line. The compound, however, has not reached the criteria of significant anti-HIV activity.

Animals↗

Evaluation of toxicity of cypenhymustine, a new anticancer compound, in mice.

The toxicity of cypenhymustine, a potential anticancer compound 1 (Cancer Letters, 70 (1993) 1-6), was assessed in normal as well as in Ehrlich ascites carcinoma (EAC), Sarcoma-180 (S-180) and Dalton's lymphoma (DL)-bearing Swiss male mice by measuring drug-induced changes in (1) hematological parameters and (2) femoral bone marrow cellularity on day 9 following drug treatment at the optimum dose of 3.0 mg/kg body weight from days 1 to 7. Detailed studies were also made by noting sequential changes in the above parameters in normal and EAC-bearing mice on days 12, 15, 18 and 21, respectively. The results indicate that the compound did not adversely affect hematopoiesis. From the sequential studies, it was observed that after a mild initial decrease in hematological counts, particularly in EAC-bearing treated mice, normalcy was reached within 11-14 days after termination of drug therapy. Drug induced hepatotoxicity and nephrotoxicity were also sequentially evaluated in normal and EAC-bearing mice on days 9, 12 and 15 but no such toxicities were detected. Also, body weight, skin and hair texture, and behavioural pattern (food and water intake and activity) did not reflect any toxic reaction in the host mice at this optimum dose.

Alanine Transaminase↗