Biomedical subjects

D Banerjee

Publications and source records attributed to D Banerjee.

At least 55 records · Page 3Linked to original sources

PubMed2001

Characterization and drug sensitivity of four newly established colon adenocarcinoma cell lines to antifolate inhibitors of thymidylate synthase.

Four new cell lines were established from the primary tumors of patients with untreated colorectal adenocarcinoma. Drug sensitivity and characterization of these cell lines was performed. Three of the four cell lines formed colonies in soft agar and all were tumorigenic in nude mice. The cell lines were morphologically similar but had differences in growth characteristics. Two of the cell lines, C18 (CCCL-4) and C29 (CCCL-6), had a longer doubling time compared with C85 (CCCL-1) and C86 (CCCL-2). The C18 and C29 cell lines had chromosome 17 abnormalities and evidence by immunohistochemistry of a mutant p53 and had decreased levels of thymidylate synthase and dihydrofolate reductase proteins, associated with decreased thymidylate synthase catalytic activity in C18 and no detectable activity in C29. Raltitrexed and GW1843U89 showed potent cytotoxic activity and all four cell lines displayed similar cytotoxicity to these folate thymidylate synthase inhibitors. The C18 and C29 cell lines were in general resistant to the other agents tested (methotrexate, 5-fluorouracil, nolatrexed) when compared with the C85 and C86 cell lines. These new cell lines may be useful for the study of colorectal adenocarcinoma and for evaluating new drugs or treatment schedules.

Adenocarcinoma↗

PubMed2001

Primary low-grade B-cell lymphoma of mucosa-associated lymphoid tissue type arising in the urinary bladder: report of 4 cases with molecular genetic analysis.

CONTEXT: Primary lymphoma of the urinary bladder is rare. Only 84 cases have been reported in the English literature to date, and none of these cases has had molecular confirmation of clonal immunoglobulin gene rearrangement. OBJECTIVES: To review all cases with primary urinary bladder lymphoma in our records, to classify them using the REAL classification, to confirm their immunophenotype and genotype, and to determine their outcome. DESIGN: We identified 4 cases of primary urinary bladder lymphoma in our medical records from a 30-year period. Immunohistochemical detection of immunoglobulin light chains and molecular analysis of immunoglobulin heavy-chain genes using the polymerase chain reaction were performed on paraffin-embedded material. RESULTS: All patients were older than 60 years. The male-female ratio was 1:3. All patients had a history of chronic cystitis. Histologic features of mucosa-associated lymphoid tissue lymphoma with centrocyte-like cells, plasmacytoid B cells, or both were observed in all cases. Monoclonality of B cells was demonstrated by immunohistochemistry, polymerase chain reaction, or both methods in every case. All patients presented with stage IAE disease, were treated with radiotherapy alone, and have been in continuous complete remission for 2 to 13 years. CONCLUSIONS: Primary bladder lymphomas are usually of low-grade mucosa-associated lymphoid tissue type. They are more common in females and are associated with a history of chronic cystitis. Lymphoepithelial lesions are seen only in association with areas of cystitis glandularis. B-cell clonality is readily demonstrable by immunohistochemistry and/or polymerase chain reaction analysis. Local radiotherapy appears to confer long-term control.

Aged↗

PubMed2001

Growth factors and stromal matrix proteins associated with mammographic densities.

Extensive radiologically dense breast tissue is associated with a marked increase in breast cancer risk. To explore the biological basis for this association, we have examined the association of growth factors and stromal matrix proteins in breast tissue with mammographic densities. Ninety-two formalin-fixed paraffin blocks of breast tissues surrounding benign lesions were obtained, half from breasts with little or no density and half from breasts with extensive density, matched for age at biopsy. Sections were stained for cell nuclei, total collagen, the stromal matrix regulatory protein tissue metalloproteinase-3 (TIMP-3), and the growth factors, transforming growth factor-alpha and insulin-like growth factor (IGF-I). The area of immunoreactive staining was measured using quantitative microscopy. Breast tissue from subjects with extensive densities had a greater nuclear area (P = 0.007), as well as larger stained areas of total collagen (P = 0.003), TIMP-3 (P = 0.08), and IGF-I (P = 0.02) when compared with subjects with little breast density. Differences were greater for subjects less than 50 years of age. These data indicate that increased tissue cellularity, greater amounts of collagen, and increased IGF-I and TIMP-3 expression are found in tissue from mammographically dense breasts and suggest mechanisms that may mediate the associated increased risk of breast cancer.

Age Factors↗

PubMed2001

Obesity predisposes to increased drainage following axillary node clearance: a prospective audit.

BACKGROUND: Whilst sentinel node biopsy is being evaluated for optimising treatment of the axilla, axillary dissection remains the gold standard. Seroma formation, a common sequel to axillary dissection, has been shown to be associated with an increased incidence of wound infection, delayed healing, and lymphoedema. This study was conducted to evaluate the possible contributory role of obesity in axillary drainage following lymphatic dissection. PATIENTS AND METHODS: This study comprised a prospective review of all patients undergoing axillary dissection in conjunction with mastectomy or wide local excision. The total in-patient axillary drainage and the average daily drainage was correlated with various clinical parameters, including obesity, type of surgery, level of axillary dissection and nodal involvement. The body mass index (BMI) was used as a measure of obesity. RESULTS: During a 6-month period, axillary dissection was performed in 79 women. Nineteen patients were excluded. Patey mastectomy was performed on 33 (55%) and the remaining had breast conservation. The amount or duration of axillary drainage did not correlate with the type of operation, tumour histology, level of axillary dissection or the nodal status. Higher BMI correlated with increased mean daily axillary drainage and total volume drained, whilst in hospital. (Spearman correlation coefficient 0.42; P < 0.01). CONCLUSION: Obesity predisposes to increased axillary drainage following nodal clearance.

Adult↗

PubMed2001

Genasense (Genta Inc).

Genasense (formerly known as G-3139), an antisense oligonucleotide specific for Bcl-2, is under development by Genta as an iv drip infusion for the potential treatment of various cancers including melanoma, prostate, breast and colon cancer [3083751. It is in phase III trials for malignant melanoma, for which it has been awarded Fast Track status 1359044]. Genasense received Orphan Drug status in August 2000 [3782331. In September 2000, the company announced that pivotal phase III trials in multiple melanoma, chronic lymphocytic leukemia (CLL) and acute myelocytic leukemia (AML) would be underway by 2001 [382783]. By January 2001, trials in AML and CLL had been initiated 1396512]. As of February 2001, Genta was planning the initiation of two additional, registration quality trials. Pending positive results from these trials, launch of Genasense is anticipated in 2002 13984111. A phase III trial in patients with advanced multiple myeloma at 65 centers in the US, Canada and Great Britain began in February 2001. The trial will examine whether the addition of Genasense can improve response rates, response duration and quality of life compared with dexamethasone therapy alone 13989081. Genta Inc has been issued a patent (US-05831066) for Genasense 1283005]. The patent provides protection to Genta for the composition of Genasense and its analogs. Furthermore, Genta Inc has also been issued two new patents that cover a series of compounds containing new backbone constructions that enhance the antisense affinity of the drug to the target pre-RNA, while the other patent covers the methods for preparation of antisense oligonucleotides containing the new backbone structures 12896851. Genta has already licensed the rights for the use of Bd-2 as a target for antisense- and gene therapy-based treatments from The University of Pennsylvania. The licensing agreements with Chugai Pharmaceutical Co for worldwide marketing and profit sharing places Genta in a favorable position. In January 2001, Needham & Co expected Genasense to have a potential market of 47,700 malignant melanoma patients in the US. The analysts also expected potential patient market sizes of 50,000 (CLL), 21,000 (AML), 136,000 (non-small cell lung cancer; NSLCC) and 180,000 (prostate cancer) in the US. In addition, the analysts predicted that Genasense would be approved for melanoma in the second quarter of 2002, with approvals to follow for CLL (third quarter of 2002), AML (third quarter of 2002) and myeloma (fourth quarter of 2002) 1399251].

Antineoplastic Agents↗

PubMed2001

Sequence-dependent enhancement of cytotoxicity produced by ecteinascidin 743 (ET-743) with doxorubicin or paclitaxel in soft tissue sarcoma cells.

Ecteinascidin 743 (ET-743) is a potent antitumor agent from the Caribbean tunicate Ecteinascidin turbinata and is presently in clinical trials for human cancers. To better understand how ET-743 might be used clinically, the present study used SRB assays to examine the cytotoxicity resulting from combining ET-743 with three other antineoplastic agents: doxorubicin (DXR), trimetrexate, and paclitaxel in different administration schedules in two soft tissue sarcoma cell lines, HT-1080 and HS-18, in vitro. Concurrent exposure of ET-743 with DXR resulted in synergistic interactions in both cell lines. Addition of ET-743 for 24 h before DXR was the most effective cytotoxic regimen against both cell lines. Morphological study by fluorescence microscopy revealed that combination treatment of both cells with ET-743 and DXR induced apoptosis. Exposure to paclitaxel before ET-743 was also an effective regimen. These results encourage studies of the combination of ET-743 and DXR in the treatment of soft tissue sarcoma, because each of these agents have activity in this disease.

Antineoplastic Agents↗

PubMed2001

Cholelithiasis in a child--an unusual presentation of Wilson's disease.

A nine year old mentally retarded girl with moderate splenomegaly and ascites presented with chronic cholelithiasis. The presence of Kayser-Fleischer rings and low serum ceruloplasmin level confirmed the diagnosis of Wilson's disease. Cirrhosis of liver and recurrent episodes of hemolysis--these two common complications of Wilson's disease make an ideal setting for gall stone formation. Only three such cases have been reported worldwide and ours is the first case report from India. We suggest that cholelithiasis and splenomegaly in a child without evidence of congenital hemolytic disease should be taken as a suspect of Wilson's disease.

Child↗

PubMed2000

Intranasal vaccination of mice against infection with Mycobacterium tuberculosis.

The intranasal (i.n.) route of immunisation, has recently been of active interest in endeavours to improve the efficacy of vaccination against a number of respiratory infections. Here, we examined the outcome of tuberculous infection in BALB/c mice. I.n. application of the BCG-Pasteur strain was found to be highly protective against challenge infection with the pathogenic H37Rv strain given after a 4-week interval, reflected by the 100-fold reduction of CFUs in both lungs and spleens. Vaccination with the recombinant PstS-1 antigen and cholera toxin significantly protected against the challenge given 10 days later, but only marginally after 12 weeks. Histological examination showed, that i.n. vaccination abrogated the confluent infiltration of lungs with inflammatory cells, which surrounds the granulomas in H37Rv challenged control mice. In conclusion, the strong protection demonstrated by BCG suggests that the i.n. route of vaccine delivery deserves further attention toward improving vaccination against tuberculosis.

Administration, Intranasal↗

PubMed2000

Identification of a glialblastoma cell differentiation factor-related gene mRNA in human microvascular endothelial cells.

Vascular endothelial cells (VEC) transduce mitogenic and chemoattractant signals in response to erythropoietin (Epo). An analysis of changes in gene expression in VEC would be helpful to understanding the molecular nature of mitogenic signals. An effective method for analysis of gene expression is through differential display. Using this approach, we obtained from Epo-treated human microvascular endothelial cells (HMVEC) a cDNA fragment with characteristics of the 3'end of mRNA. Using the cDNA fragment, we then isolated a full-length clone from a HMVEC cDNA library. The cDNA of interest encodes a protein consisting of 404 amino acids with a carboxy-terminal end sequence identical to glialblastoma cell differentiation factor-related protein (GBDR1). Northern blot analysis showed that GBDR1 mRNA was ubiquitously expressed in human tissues. In Southern blot analysis, GBDR1 cDNA identified a single gene on chromosome 9. Since analysis of the amino acid sequence revealed several putative phosphorylation sites for different protein kinases, the GBDR1 protein was expressed and purified from bacterial extracts and, as predicted, casein kinase II phosphorylated GBDR1 in vitro. Immunofluorescence and biochemical data revealed that the GBDR1 protein is not entirely localized in the cytosolic fraction, suggesting that it may interact with another protein(s). These findings demonstrate that GBDR1 is an intracellular signaling molecule that may play a role in the regulation of endothelial cell growth.

Amino Acid Sequence↗

PubMed2000

Levels of E2F-1 expression are higher in lung metastasis of colon cancer as compared with hepatic metastasis and correlate with levels of thymidylate synthase.

We recently reported that forced overexpression of the transcription factor E2F-1 in human HT-1080 fibrosarcoma cells resulted in corresponding high levels of thymidylate synthase (TS) and resistance to 5-fluoropyrimidines (D. Banerjee et al., Cancer Res., 58: 4292-4296, 1998). Because colorectal metastasis to the lung has higher TS levels than liver metastasis and is less responsive to treatment with 5-fluorouracil (R. Gorlick et al., J. Clin. Oncol., 16: 1465-1469, 1998), it was, therefore, of interest to measure E2F-1 expression in these tumors. In contrast to marginally increased levels of dihydrofolate reductase and topoisomerase I in lung metastasis as compared with liver metastasis, lung tumors had a 5-fold increase in E2F-1 expression as compared with liver tumors, corresponding to the relative levels of TS in these metastases. These data indicate that there exists a close correlation between E2F-1 and TS levels and provide a rationale for targeting this transcription factor, ie., E2F-1, for the treatment of certain cancers.

Adenocarcinoma↗

PubMed2000

Cloning and characterization of complementary DNA encoding human N-acetylglucosamine-phosphate mutase protein.

Endothelial cells express erythropoietin receptor (EpoR) and are responsive to erythropoietin (Epo). Upon ligand binding, EpoR activates multiple signaling cascades. Identification of genes expressed in response to Epo is important for understanding the molecular nature of the signals. Applying the differential display approach, an effective method for analysis of gene expression, we identified five differentially expressed mRNAs. In this study, we cloned human N-acetylglucosamine-phosphate mutase from a human microvascular endothelial cell (HMVEC) cDNA library using one of the differentially expressed fragments as a probe. The nucleotide (nt) sequence analysis of the longest clone displayed a 2 kb cDNA fragment and encodes a protein of approximately 542 amino acids with a predicted MW of approximately 60 kDa. Northern blotting and reverse transcriptase-polymerase chain reaction analysis revealed an upregulation of the N-acetylglucosamine-phosphate mutase mRNA after 2 h of stimulation of cells with Epo. This gene was shown to be variably expressed in human tissues and is located on chromosome 6. These studies demonstrate that the expression of N-acetylglucosamine-phosphate mutase mRNA responds to cytokines, and the presence of a 10 aa motif similar to the putative active site of several hexose-phosphate mutases provides a basis for future studies of the role of this gene in the regulation of Epo-stimulated endothelial cell proliferation.

Amino Acid Sequence↗

PubMed2000

Gene therapy for endometrial carcinoma with the herpes simplex thymidine kinase gene.

We investigated whether retrovirus-mediated transfer of the herpes simplex thymidine kinase gene into a human endometrial carcinoma (EC4) cell line can sensitize these cells to the prodrug ganciclovir (GCV) and thereby provide a therapeutic option for this cancer. A retrovirus encoding for the herpes simplex virus tip-1 (HSV) thymidine kinase (tk) gene was generated in which expression of tk is under control of the myeloproliferative sarcoma virus (MPSV) promoter/enhancer. We used human mutated dihydrofolate reductase (DHFR) cDNA as a selectable marker. Expression of tk was confirmed by Northern blot analysis and reverse transcription polymerase chain reaction. We demonstrated that the combination of retrovirally mediated tk gene transfer and GCV treatment effectively inhibits proliferation and causes death of EC4 cells in vitro. A bystander killing effect was observed when 90% of uninfected tumor cells were mixed with only 10% of HSVtk-infected cells. We suggest that a gene therapy approach to endometrial carcinoma can be established using retroviral transfer of HSVtk to tumor cells and subsequent administration of GCV.

Antiviral Agents↗

PubMed2000

Intestinal inflammation observed in IL-2R/IL-2 mutant mice is associated with impaired intestinal T lymphopoiesis.

BACKGROUND & AIMS: Although interleukin (IL)-2(-/-) and IL-2Ralpha(-/-) mice develop inflammatory bowel disease, IL-2Rbeta(-/-) animals are apparently free of gut pathology. Intraintestinal T lymphopoiesis is reported to be impaired in IL-2Rbeta(-/-) mice; we have determined whether this characteristic correlated with the apparent resistance of this mutant strain to intestinal inflammation. This led us to reassess intraintestinal T lymphopoiesis in these 3 mutant strains. METHODS: Intestinal histology and intraintestinal T lymphopoiesis were analyzed in unmanipulated mutant mice and in athymic and euthymic radiation chimeras reconstituted with bone marrow derived from IL-2(-/-), IL-2Ralpha(-/-), and IL-2Rbeta(-/-) donors. RESULTS: Intraintestinal T lymphopoiesis was ablated in the 3 mutant strains and was associated with cryptopatch abnormalities. The intestinal mucosa of mice reconstituted with lymphocytes from IL-2Rbeta(-/-) mice exhibited lesions of both the small and large bowel similar to those observed in the early stages of human gluten enteropathy and acute ulcerative colitis, respectively. Analysis of euthymic and athymic bone marrow radiation chimeras indicated that T cells located in the intestinal mucosa of unmanipulated IL-2(-/-), IL-2Ralpha(-/-), and IL-2Rbeta(-/-) mice are of thymic origin. CONCLUSIONS: Null mutations at IL-2/IL-2Ralpha and beta loci differentially affect intraintestinal and intrathymic T lymphopoiesis. These conditions are associated with lesions of intestinal inflammation that are mediated by thymus-derived T cells.

Animals↗

PubMed2000

Iris lymphoma in a pediatric cardiac transplant recipient: clinicopathologic findings.

PURPOSE: To report the unusual presentation of a confirmed iris lymphoma in the setting of posttransplant lymphoproliferative disorder. DESIGN: Case report with clinicopathologic findings. METHODS: The medical record was reviewed for clinical manifestations, course of disease, and pathologic and radiologic findings. RESULTS: A 4-year-old girl who had undergone a cardiac transplant at 3 months of age was found to have unilateral iris lesions associated with granulomatous uveitis. Her condition was unresponsive to local and systemic steroids, so she underwent an iris biopsy that was found to contain a B cell-derived monoclonal lymphoproliferation. Treatment with systemic chemotherapy including cyclophosphamide and prednisone resulted in a transient, partial response. Localized radiotherapy induced total regression of the iris lesions. She continues to be observed and has been free of recurrence or metastasis for the past 12 months. CONCLUSIONS: Because of the increased use of organ transplantation, eye care specialists should consider the diagnosis of intraocular lymphoma when confronted with uveitis-like syndrome unresponsive to topical steroid therapy. Although there are numerous treatment options for intraocular lymphoma, radiation therapy was found to be effective in the treatment of this case.

Antineoplastic Combined Chemotherapy Protocols↗

PubMed2000

Urethral carcinoma in women: results of treatment with primary radiotherapy.

BACKGROUND AND PURPOSE: Urethral carcinoma in women is uncommon. This study was undertaken to evaluate the role of radiotherapy in the treatment of these tumors. MATERIALS AND METHODS: The hospital records of 34 women with primary urethral carcinoma were retrospectively reviewed. There were 15 squamous cell carcinomas, 13 transitional cell carcinomas, and six adenocarcinomas. The primary tumor was >4cm in size in eight patients, involved the proximal urethra in 19 and extended to adjacent organs in 22. Inguinal or iliac lymphadenopathy was present in nine patients. There were eight TNM stage I/II tumors, 11 stage III tumors and 15 stage IV tumors. Radiotherapy was administered only to the primary tumor in 15 patients, and to the primary tumor and regional lymph nodes in the remaining 19 patients. Brachytherapy with or without external radiation was used to treat the primary tumor in 20 patients. RESULTS: Tumor recurred in 21 patients. The 7-year actuarial overall and cause-specific survivals were 41 and 45%, respectively. Large primary tumor bulk and treatment with external beam radiation alone (no brachytherapy) were independent adverse prognostic factors for local tumor recurrence. Brachytherapy reduced the risk of local recurrence by a factor of 4.2. The beneficial effect of brachytherapy was most prominently seen in patients with bulky primary disease. Large tumor size was the only independent adverse predictor of overall disease recurrence and death from cancer. CONCLUSIONS: Radiotherapy is an effective treatment for carcinoma of the female urethra and preserves normal anatomy and function. Brachytherapy improves local tumor control, possibly as a result of the higher radiation dose that can safely be delivered.

Adenocarcinoma↗

PubMed2000

Retrospective dosimetry: estimation of the dose to quartz using the single-aliquot regenerative-dose protocol.

We report on the application of the single-aliquot regenerative-dose protocol to retrospective dosimetry, using the optically stimulated luminescence (OSL) from quartz extracted from fired bricks. These bricks had previously been exposed to enhanced levels of ionising radiation while part of inhabited structures in the Chernobyl area. The time dependence of the OSL signals is considered first and we conclude that it is most appropriate to use the initial part of the OSL signal for dose estimation, after subtraction of a slowly varying background component. It is then shown that this signal meets the fundamental requirement of the single-aliquot regenerative-dose protocol, in that any change in the luminescence recombination probability can be corrected for by using the OSL response to a fixed test dose. The response of a particular aliquot is examined after three different treatments (untreated, reset by exposure to light and reset by heating to 500 degrees C) and it is shown that, after sensitivity correction, the dose-response curves are indistinguishable up to 10 Gy. The routine application of the protocol is then described and dose estimates are shown to be insensitive to preheat temperature and test-dose size. Finally, dose-depth profiles are presented for two bricks. These profiles demonstrate that the high precisions (approximately 1%) obtained using the regenerative-dose protocol are reflected in smooth dose-depth dependencies.

Construction Materials↗

PubMed2000

Studies of solvation of ketocyanine dyes in homogeneous and heterogeneous media by UV/Vis spectroscopic method.

Solvation characteristics of ketocyanine dyes (I-VI) have been investigated in pure solvents and heterogeneous media by absorption and fluorescence studies. The dyes are good reporters of solvent polarity. In protic solvents they exist as equilibrium mixtures of bare and hydrogen-bonded form in the ground state (S0), the latter being the emitting species. In aprotic solvents of low polarity association of the S1 state of the dye takes place. In aqueous micellar media the dye resides at the micelle water interface. The binding constant for dye-micelle interaction has been determined. Fluorescence data in beta-cyclodextrine solution resemble that for that neutral micellar solution indicating that the interaction between the -OH group of the heterogeneous part (micelle/cyclodextrine cavity) and the carbonyl oxygen of the dye is important in both the cases.

Alkenes↗

PubMed2000

Comparison of methotrexate resistance conferred by a mutated dihydrofolate reductase (DHFR) cDNA in two different retroviral vectors.

We previously reported the protection of hematopoietic cells from methotrexate (MTX) toxicity using an N2-based double copy vector containing serine 31 (S31)-mutated dihydrofolate reductase (DHFR) (DC/SV6S31). To examine whether the use of SFG-based dicistronic vectors will lead to improvement in gene transfer over the DC/SV6 vector, we compared the protection provided by MTX to NIH3T3 cells and hematopoietic progenitor cells infected with these retroviral constructs containing the S31 variant DHFR cDNA. In NIH3T3 cells, the 50% effective dose values of MTX conferred by the SFG vector were 8-fold higher than those obtained with the DC/SV6 vector. DHFR mRNA levels were 22-fold and 38-fold higher than that seen for the DC/SV6 vector according to Northern blot and real-time polymerase chain reaction analysis, respectively. However, DHFR protein expression and DHFR enzyme activity were only 1.5-fold and 2-fold higher in the SFG vector, respectively, indicating that the mRNA from the SFG vector is translated less efficiently than the mRNA generated from the DC/SV6 vector. Furthermore, the degree of MTX protection conferred by each vector in both mouse and human hematopoietic cells was the same. These results indicate that the in vitro transduction efficiency and transgene expression of human DHFR in hematopoietic progenitor cells is equally conferred by both vectors.

3T3 Cells↗