Erratum: Theory of a two-photon squeezed laserlike oscillator
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Biomedical subjects
Publications and source records attributed to R Ghosh.
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The purpose of this work was to compare the efficacy of immunizing mice with a soluble vs a cell-associated form of a tumor Ag. A murine B cell tumor (2C3), which displays an Id on its cell surface, grows progressively and gives rise to Id-negative tumor variants in nonimmunized animals. We previously reported that the tumor variants arise as a consequence of Id-specific T cell suppression of the Id+ tumor. The Id-specific effector T cells are CD4+, CD8-. Based upon these findings vaccination protocols have been designed and tested to determine whether the expansion of tumor-specific effector T cells would eliminate the Id+ tumors and prevent the subsequent generation of Id- tumor variants in vivo. MHC-restricted T cells typically recognize soluble Ag subsequent to modification by an APC, and APC may ultimately express a processed form of the Ag that is different from that expressed on the surface of the tumor cells. Based upon this assumption, the efficacy of immunizing mice with cell-associated 2C3 Id was compared to immunization with a soluble form of the same Id. Mice were immunized with either irradiated 2C3 cells or syngeneic spleen cells to which 2C3 protein was covalently linked. These immunization protocols provided a complete and lasting protection against a tumor challenge of up to 1 x 10(6) tumor cells. In contrast, most mice hyperimmunized with a soluble form of the Id did not survive this level of tumor challenge in spite of the production of significant levels of anti-Id antibodies. Mice immunized with the soluble form of the Id, which did survive, produced slowly progressing tumors expressing a 1000-fold less of the marker Id. These results illustrate the importance of understanding and properly exploiting a host's natural response to a tumor-specific Ag when designing effective immunization protocols for cancer therapy.
The incidence of sister chromatid exchange (SCE) was investigated in the lymphocyte chromosomes of 45 patients with oral submucous fibrosis and 56 age- and sex-matched nonsmoking controls. The frequency of SCE was 9.26 +/- 2.15 in patients with oral submucous fibrosis, which was significantly higher than the mean SCE value of 5.49 +/- 1.24 observed in normal controls. The frequency of SCE in patients with oral submucous fibrosis addicted to the habit of betel with tobacco chewing, "bidi"/cigarette smoking and combined habits of chewing and smoking of tobacco were 8.12 +/- 1.69, 9.43 +/- 1.87, and 10.06 +/- 2.28, respectively. These values were also significantly higher as compared with the SCE values observed in normal controls.
Cytogenetic analysis was performed in a young girl with breast carcinoma. The most important finding was the existence of double minute (dmin) chromosomes, besides other chromosomal abnormalities. Removal of the tumor resulted in reduction of chromosomal abnormalities in the leukocyte cultures repeated after 1 year, suggesting that the presence of tumor may elevate the frequency of chromosomal abnormalities. The presence of dmin chromosomes indicates that factors causing their presence in tumor cells may also operate in human leukocytes.
A recently described collagen gel culture technique has been modified to evaluate the growth characteristics and chemosensitivity patterns of genitourinary neoplasms. Fresh human surgical explants incorporated radiolabeled DNA precursors [H3)thymidine or deoxyuridine) in 97% of 38 patient specimens (18/19 bladder, 3/3 prostate, 2/2 testis, 13/14 renal), after being maintained for three to 13 weeks/passage, with several specimens reaching their sixth passage (20 months). Control cellular DNA incorporation ranged from five to 90% (#cells labeled/#cells evaluated), with median labeling for bladder 30%, prostate 80%, testis 90%, and renal 80%. Original histopathologic classification was maintained in all cases. Tumor volume and glucose consumption were other measurable parameters. Seventy-three surgical specimen cultures were treated with chemotherapeutic agents after a minimum of four weeks in culture. Single agent exposures were 24 hours at 1X and 10X reported peak plasma concentrations. Combination agents were sequenced as in current clinical protocol for bladder tumors and fourteen day continuous fluorodeoxyuridine (FdURD) exposure for renal tumors. Sensitivity was found in 1/2 prostate and 1/10 renal tumors to Adriamycin, 8/15 bladder and 1/2 testis tumors to cisplatin, 11/28 renal tumors to FdURD and 6/16 bladder tumors to MVAC combination chemotherapy. This culture system offers the advantages of in vivo-like solid tumor growth, a high culture success rate, longevity in culture, maintenance of the primary histopathology and reproducible chemosensitivity response.
Reported here are studies of Fanconi anemia fetal cells that led to the first use of umbilical cord blood for hematopoietic reconstitution in a clinical trial. Prenatal diagnosis and HLA typing were performed in fetuses at risk for Fanconi anemia (FA) to identify, prior to birth, those that were unaffected with the syndrome and were HLA-identical to affected siblings. Umbilical cord blood was harvested at the delivery of these infants; assays of progenitor cells indicated the presence of colony-forming units-granulocyte-macrophage (CFU-GM) in numbers similar to those of bone marrow CFU-GM that are associated with successful engraftment in HLA-matched allogeneic bone marrow transplantation. The possibility that umbilical cord blood from a single individual can be used as an alternative to bone marrow for hematopoietic reconstitution has now been demonstrated by the successful engraftment of two patients with FA. Progenitor cell assays of umbilical cord blood collected at the birth of a child affected with FA, who had been misdiagnosed on the basis of chorionic villus sampling (CVS) studies, indicated a profound deficiency in colony formation, consistent with previously reported abnormalities in the growth of FA cells in vitro. These results suggest that the hematopoietic disorder in FA is related to an underlying problem with cell proliferation.
Involvement of topoisomerase II in the repair of damage by N-methyl-N'-nitro-N-nitrosoguanidine, UV light and gamma-rays has been studied using quiescent V79 cells. In each case, the presence of nalidixic acid, the inhibitor of topoisomerase II, decreased the survival through suppression of potentially lethal damage repair. There was also an increase in the yield of mutants because of such suppression. The observations were in contrast with those made with exponential cells in the following aspects: (a) density-inhibited cells showed a positive response to suppression of topoisomerase II activity after treatment with UV light and gamma-rays and (b) for MNNG exposure, mutational yield increased instead of decreasing as in exponential cells. The results showed that topoisomerase II played an important part in the repair of damage of density-inhibited cells.
Distamycin-A, an oligopeptide antibiotic with a N-methylpyrrole ring system and propionamide side chain, preferentially forms stable bonds with AT rich double stranded DNA. When introduced to cell cultures, it inhibits condensation of the heterochromatic region of the Y chromosome. The frequency of metaphases showing inhibition of heterochromatin condensation of the Y chromosome was found to be dependent on the treatment time and concentration of distamycin-A in the culture medium. When distamycin-A was added to a concentration of 100 micrograms/ml at the start of the culture (72 hours), the frequency of Y heterochromatin decondensation was found to be 48%, 30% and 6% in amniotic fluid, lymphocyte and fibroblast cultures respectively. The highest frequency of metaphases with decondensed Y heterochromatin were observed when distamycin-A treatment was carried out for the last 24 hours prior to harvest, the frequencies being 94%, 72% and 59% in amniotic fluid, lymphocyte and fibroblast cultures respectively. Increase in the concentration of distamycin-A from 25 micrograms/ml to 50 micrograms/ml during the last 24 hours of culture increased the incidence of metaphases with Y heterochromatin decondensation from 51% to 69% in amniotic fluid, 40 to 49% in lymphocyte and 29% to 31% in fibroblast cultures. Highest frequency of metaphases with Y heterochromatin decondensation were observed when the cultures were exposed to distamycin-A at a concentration of 100 micrograms/ml for the last 24 hours of culture.
C21H23N2O2+.Cl-.1.5H2O, Mr = 397.90, monoclinic, P2(1), a = 7.617 (1), b = 32.463 (2), c = 7.849 (1) A, beta = 90.40 (2) degrees, V = 1940.8 (4) A3 Z = 4, Dm = 1.39, Dx = 1.36 g cm-3, lambda (Cu K alpha) = 1.5418 A, mu = 19.724 cm-1, F(000) = 844, final R = 0.047 for 3368 observed reflections. Strychnine hydrochloride is a naturally occurring alkaloid in its protonated form. One of the N atoms accepts the proton from hydrochloric acid changing its hybridization to sp3. The indole N atom is sp2 hybridized with its lone-pair electron involved in conjugation with the carbonyl group. The hydrogen bonds form a complex zigzag network which stabilizes the structure.
The epitope reactivities of 52 well-characterized monoclonal antibodies (Mabs) against carcinoembryonic antigen from 11 different research groups were studied using competitive solid-phase immunoassays. About 60% of all possible combinations of Mabs as inhibitors and as the primary binding antibody were investigated. The inhibition data were analyzed by a specially developed computer program "EPITOPES" which measures concordance and discordance in inhibition patterns between Mabs. The analysis showed that 43 of the 52 Mabs (83%) could be classified into one of five essentially noninteracting epitope groups (GOLD 1-5) containing between four and 15 Mabs each. The epitopes recognized by the Mabs belonging to groups 1 to 5 were peptide in nature. With one or two possible exceptions non-classifiable Mabs were either directed against carbohydrate epitopes (4 Mabs) or were inactive in the tests used. Within epitope groups GOLD 1, 4, and 5 two partially overlapping subgroups were distinguished. Mabs with a high degree of carcinoembryonic antigen specificity generally belonged to epitope groups GOLD 1 and 3.
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To investigate whether anti-(carcinoembryonic antigen) monoclonal antibodies (mAb) react with single or repeated epitopes, sandwich radioimmunoassays in homologous and heterologous combinations were performed. Four mAb (I-27, I-47, II-17 and to some degree II-16) gave homologous binding while two mAb (I-38S1 and II-10) did not. Taken together with previous immunoprecipitation studies we conclude that all these mAb except II-10 react with repeated epitopes. The relative positions of the epitopes recognized by these mAb and of three additional mAb (II-6, II-7 and CB-CEA-1) were investigated using a plate antibody competition test with enzyme-labelled carcinoembryonic antigen (CEA). mAb I-38S1, II-6, II-7, II-10, II-16 and CB-CEA-1 were mutually cross-reactive, and were classified as belonging to one epitope group. mAb I-27 and I-47 fell outside this group and did not interfere with the binding of CEA conjugate to mAb II-17 either. They therefore represent a second epitope group. mAb II-17 showed no interference with the binding of CEA to any of the other mAb and must therefore represent a third epitope group. The slopes of the plate antibody competition curves were used for calculation of a correlation matrix, which in turn was used to depict the relative positions of the epitopes recognized by the mAb in the large group.
Density-inhibited V79 cells when held for 24 h in complete medium after exposure to N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) show improved survival levels and decreased mutant frequencies at all dose levels, compared to cells not so held. However, when benzamide, an inhibitor of poly(ADP-ribose) synthesis was present during this 24-h holding, the improvement in survival and decrease in mutant frequencies were not observed. Rather, compared to the control, the cells became more sensitive to MNNG and mutant frequency also increased significantly for all doses studied.
V-79 cells when exposed to thymidine (5 micrograms/ml) in growth medium after treatment with X-rays, UV light and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), responded differently depending upon the agent. For treatment with X-rays and UV light, only induction of mutation was potentiated, but for MNNG treatment, both killing and mutation induction were potentiated. The increase in killing of MNNG exposed cells could be reversed by simultaneous addition of deoxycytidine with thymidine, but, for all the three mutagenic treatments, enhancement in mutation induction could not be suppressed by deoxycytidine.
The structural and motional properties of mixed bilayers of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) have been examined by using wide-line 31P, 14N, and 2H NMR. 2H and 14N NMR data showed that in mixed bilayers containing both PC and PE the conformations of the head-group moieties are essentially identical with those observed for bilayers containing a single phospholipid species. Equimolar amounts of cholesterol induce also only a small change in head-group conformation. 31P T1 relaxation measurements (at 300 MHz) at various temperatures of bilayers containing phospholipids with a mixture of phosphocholine and phosphoethanolamine head-groups and unsaturated fatty acid residues revealed in all cases a clearly defined minimum corresponding to the condition omega O tau C-1 approximately 1. For all phospholipid mixtures studied, the 31P T1 relaxation was homogeneous over the whole powder spectrum and could be fitted to a single-exponential decay. The 31P vs temperature profiles were analyzed by a simple correlation model following the analysis of Seelig et al. (1981) [Seelig, J., Tamm, L., Hymel, L., & Fleischer, S. (1981) Biochemistry 20, 3922-3932]. Rotational diffusion of the phosphate moiety in bilayers of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) was slower than that of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and the activation energy was increased by a factor of 1.7 to 31.4 kJ mol-1.(ABSTRACT TRUNCATED AT 250 WORDS)
The incidence of sister chromatid exchange (SCE) was investigated in the lymphocytes of control women, pregnant women, and women using oral contraceptives after culture at 37 degrees C and 40 degrees C. At 37 degrees C, the mean frequency of SCE (MEAN +/- S.E.) was found to be 7.91 +/- 0.30 in pregnant women and 8.53 +/- 0.29 in oral contraceptive users which were significantly higher than the SCE value of 5.56 +/- 0.21 found in control women. Increase in growth temperature to 40 degrees C elevated the SCE frequency to 11.86 +/- 0.44 in pregnant women, 12.76 +/- 0.46 in oral contraceptive users and 7.24 +/- 0.26 in control women. These data indicate that there is a differential induction of SCEs following increased cell culture temperature in the lymphocytes of pregnant women and oral contraceptive users, compared with control women.
The effect of treatment with nalidixic acid, an inhibitor of DNA topoisomerase, after exposure of V79 cells to different DNA-damaging agents on the induction of killing and mutation has been studied. The DNA-damaging agents were ultraviolet light, gamma-rays and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). It was seen that treatment with nalidixic acid potentiated the killing by MNNG and suppressed the induction of mutation. However, it had no influence upon killing and mutation by UV light and gamma-rays. The difference in the observed results could be due to the nature of the damage induced and its repair in relation to the function of topoisomerases.
The incidence of sister chromatid exchange (SCE) was investigated in lymphocyte chromosomes of 59 patients with oral leukoplakia and 65 age- and sex-matched nonsmoking controls. The frequency of SCE was found to be 8.61 +/- 1.89 in patients with oral leukoplakia, which was significantly higher than the mean SCE value of 5.58 +/- 1.26 observed in normal controls. The frequency of SCE in patients with oral leukoplakia addicted to the single habit of betel with tobacco chewing, bidi/cigarette smoking, and combined habits of chewing and smoking of tobacco were found to be 7.95 +/- 1.63, 8.17 +/- 1.66, and 9.23 +/- 2.14, respectively. These values were also significantly higher as compared to the SCE values observed in normal controls.