Ocular manifestations of parenteral administration of scopolamine (hyoscine).
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Glaucoma↗
Biomedical subjects
P Chandra
Publications and source records attributed to P Chandra.
[Remarks on the article by M. Reim. Experiences on the applications and stability of 5-iodouracil desoxyriboside].
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Drug Stability↗
A study of tolerance of ether anaesthesia to dogs with varying grades of chronic anaemia.
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Anemia↗
Sickle cell anemia- clinical manifestations in 100 patients and review of the literature.
The clinical and hematological features of 100 patients with sickle cell anemia are reviewed. The heart was enlarged and a murmur was heard in nearly 80 percent of patients. Pneumonia and pulmonary infarction occurred in 43 percent and 12 percent of patients, respectively. Musculoskeletal involvement included the hand-foot syndrome (15 percent), leg ulcers (55 percent), aseptic necrosis ofbone (11 percent), and osteomyelitis (4 percent). Symptoms and signs related to the gastrointestinal system included jaundice (55 percent), hepatomegaly (50 percent), splenomegaly (23 percent), hepatitis (11 percent) and gallstones (9 percent). Three patients underwent cholecystectomy and three patients had their spleens removed. Pyelonephritis occurred in 17 patients, priapism in five and hematuria in seven. Nineteen women had 39 pregnancies, of which 35 resulted in the birth of healthy infants. At least 328 painful crises occurred in 73 patients. There were also 13 hemolytic crises, eight sequestration crises, and five aplastic crises. A trail of alkali therapy in 33 crises in children failed to produce beneficial effects greater than hydration and analgesics alone as used in the control group. Laboratory findings in the 100 patients were comparable to those previously reported in the literature. The renal concentrating defect in most patients was confirmed. There were six deaths: hepatic coma secondary to post-transfusion hepatitis, thrombosis of inferior vena cava, congestive heart failure, exsanguination from erosion of the pancreaticoduodenal artery, extensive bronchopneumonia, and pulmonary infarction.
Adolescent↗
Determination of the exposure rate constant for 125I using a scintillation detector.
A scintillation spectrometer was employed to determine the exposure rate constant for an 125I seed of the type used in therapeutic radiology. The method consisted of converting counting rate to photon fluence incident upon the detector, and then calculating the exposure rate from the photon fluence. Correlations are required for absorption of the 125I photons in the air path, in the aluminum window of the NaI crystal, and in the aluminum-oxide reflector. Corrections are also required for the escape of iodine K-characteristic radiation, and the intrinsic peak efficiency of the crystal. The experimentally determined exposure rate constant in a direction perpendicular to the long axis of the seed is 1.309 Rcm2mCi-1 h-1 +/- 5.8%, and this compares favorably with a calculated value of 1.284 R cm2mCi-1 h-1. The photon fluence as a function of angle of rotation about an axis perpendicular to the seed's long axis was also measured. These data were used to estimate an average exposure rate constant for 125I seeds of 1.089 Rcm2mC-1 h-1.
Brachytherapy↗
Corneal injuries during general anesthesia.
A corneal abrasion is the most common eye complication during general anesthesia and recovery. It is painful, may progress to inflammation of the uveal tract, and in the presence of contamination may lead to a serious infection. Both eyes may be involved, as the two reported cases show. Possible causes include mask friction on the open eye or other careless technics. Treatment includes local application of an antibiotic ointment and an eye pressure patch. In addition, a cycloplegic and mydriatic solution is instilled to prevent synechiae (in the presence of a secondary iridocyclitis) and to relieve the pain associated with spasm of the iris and ciliary muscle. Topical application of local anesthetics should be avoided, as they delay regeneration of corneal epithelium and may promote keratitis.
Adult↗
Direct regulation of Na(+)-dependent myo-inositol transport by sugars in retinal pigment epithelium: role of phorbol ester and staurosporin.
An Na(+)-dependent active process for myo-inositol (MI) uptake, sharing a common carrier system with glucose and sensitive to phlorizin, was previously established in primary cultures of bovine retinal pigment epithelial (RPE) cells (26, 32). The present report further examines the nature of glucose-induced inhibition of MI transport in primary cultures of RPE cells. RPE cells were grown in supplemented Dulbecco's modification of Eagle's medium (DMEM) containing 5 mM D-glucose (basic growth media) or 40 mM D-glucose or its nonmetabolizable analogue, alpha-methyl-D-glucoside (alpha MG); 1-5 mM nonradioactive MI, pyruvate, or lactate; or 0.2-20 microM phorbol 12-myristate 13-acetate (TPA) or straurosporin (modified growth media), for up to 4 weeks. The capacity of RPE cells to accumulate 3H-MI (ratios of intracellular transported radioactive MI, [MI]i, to external free MI concentration, [MI]i/[MI]o) decreased by up to 41% or 34% when cells were grown for 10 days or longer with 40 mM D-glucose or 40 mM alpha MG, respectively, compared to cells grown in basic growth media. The rate of uptake of 3H-MI also was reduced to 63 +/- 15% or 48 +/- 8% of the control values when cells were fed 1 or 5 mM nonradioactive MI, respectively. In addition, cellular capacity to bind to [3H]phlorizin was reduced to 52 +/- 7%, 61 +/- 5%, or 38 +/- 6% of the controls when RPE cells were fed 40 mM D-glucose, 40 mM alpha MG, or 5 mM nonradioactive MI, respectively. Growth media containing either pyruvate or lactate, the glucose metabolites, did not suppress the ability of RPE cells to accumulate MI. An 18 +/- 8% reduction in [3H]thymidine incorporation into DNA occurred when cells were grown in 40 mM glucose for 12-14 days, compared to cells grown with 5 mM glucose. Chronic treatment (12-14 days) of the cells with phorbol ester, an activator of protein kinase C, caused up to twofold increase in MI uptake, [3H]phlorizin binding, cell number, and DNA synthesis. However, when the rates of MI uptake into cells grown in basic growth media or TPA-treated media were normalized to cell number, no significant difference in MI uptake was found between the treated and untreated cells. Addition of staurosporin, a protein kinase C inhibitor, together with TPA, in the growth media reversed the phorbol-induced increase of MI uptake.(ABSTRACT TRUNCATED AT 400 WORDS)
Alkaloids↗
Histone-mediated transfer and expression of the HIV-1 tat gene in Jurkat cells.
We studied the gene transfer efficiency of lipofection reagents in comparison to DEAE-Dextran. DOTAP, Dosper, and Lipofectin have lower transfection efficiency; Lipofectamine has a 2.5-fold better efficiency compared with DEAE-Dextran. We report a novel and highly efficient DNA transfer system based on the DNA-binding proteins histone 3 and histone 4. We have transferred the HIV-1 tat gene and measured the transactivation of HIV-1 LTR by the transactivator protein, expressed in Jurkat cells. The HIV-1 LTR was linked to the CAT gene as a reporter. Compared to DEAE-Dextran-mediated transfection, histone-mediated transfection resulted in a sevenfold higher expression of the CAT gene. The maximum transfection efficiency mediated by histones is dependent on the relative concentration (DNA:histone ratio) and the incubation time. In a gel-retardation assay, an optimal complex formation was observed under the same conditions that allowed the highest transfection efficiency. This ability of histones to increase the delivery and transgenic expression of foreign DNA in eukaryotic cells is not simply due to the positive ionic character of the histone proteins. Polylysine, histone H1, and histone H2A were unable to mediate gene transfection in our system. Monoclonal antibodies that recognize antigenic determinant present on all five histone proteins (anti-histone, pan) were able to neutralize the transfection-enhancing potential of histone 3 and histone 4. However, anti-histone IgG enhanced the retardation of mobility of histone-DNA complexes. The results of this study allow us to conclude that histones H3 and H4 can catalyze gene transfer and gene expression in eukaryotic cells without any requirement for additional constituents. For this reason, we have termed the new gene-delivery system as histonefection.
Animals↗
Mechanistically different acting tumor promoters induce consistent qualitative and quantitative changes in protein expression patterns of murine epidermis.
Systematic screening of different organs has shown that induction of particular proteins by 12-0-tetradecanoyl-phorbol-13-acetate is specific for mouse skin. Different compartments of skin showed all the same expression. For experimental reasons we focused the analysis performed thereafter on the epidermis. As an end- point of mouse skin tumorigenesis papilloma and carcinoma were also screened for changes in protein expression. We focused our efforts on the characterization and identification of selected, de novo expressed proteins induced by various tumor promoters in murine epidermis. The analysis presented here consists of functional characterization. Having found differences in protein expression in TPA-treated murine epidermis compared with acetone-treated control mice, we then tried to correlate the expression to the signal transduction pathway which the tumor promoter exerts its activity through. For this reason we screened protein expression patterns after topical administration of mechanistically differently acting tumor promoters, which were inflammatives not bearing tumor promoting activity anti-promoters or a combination of tumor promoters with modulators of tumor promotion. The epidermis of untreated newborn mice served as a model for high cellular proliferation and differentiation. Alterations were compared with changes observed after tumor promoter application.
Animals↗
A novel protein (p10) induced by 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and other hyperplasiogenic tumor-promoting and non-promoting agents in murine epidermis.
Treatment of murine epidermis with the tumor promotor 12-O-tetradecanoylphorbol-13-acetate (TPA) shows characteristic and significant changes in protein expression analyzed by 2D PAGE, compared to that of acetone-treated mouse epidermis. Of the seven de novo expressed proteins in TPA treated murine epidermis, one is a 44 kDa protein (p44) located nearby actin, and six proteins are in the low-molecular range between 10-20 kDa (p10, pY, pCa, p1, p2 and p3). Interestingly, the incomplete promoting and inflammative hyperplasiogen 12-O-retinoylphorbol-13-acetate (RPA) and the non-promoting but inflammative and hyperplasiogenic calcium-ionophore A23187 induced the same pattern of proteins observed in the pidermis of mice treated with TPA, with minor quantitative differences. In all cases, p10 expression was quantitative the most abundant. Partial sequencing of this protein has led to the conclusion that it is a novel protein with no such sequences in the database comparisons using FASTA and TFASTA computer programs of Genetics Computer Group. The data presented here do not strictly support the functional role of de novo induced proteins to tumor promotion, but show a causal relationship to hyperplasiogenic potency of TPA, RPA and A23187.
Amino Acid Sequence↗
Peritoneal macrophages from 17 alpha-ethinylestradiol-treated NMRI mice secrete transformation-specific low molecular weight proteins.
Analysis of protein secretion was performed for a macrophage-like cell line, which was established from the peritoneal cells of NMRI mice treated with 17 alpha-ethinylestradiol. The protein secretion pattern was investigated by computerized analysis of high resolution two-dimensional gel electrophoresis (2-D PAGE) and compared to that of control macrophages, intraperitoneally activated by bacterial lipopolysaccharide. The transformed cells encode a number of low molecular weight proteins (10-20 kDa), which were not observed in control cells under identical experimental conditions. In conclusion the transformation of peritoneal macrophages by 17 alpha-ethinylestradiol leads to an upregulation of polypeptides, in particular of low molecular weight proteins. A high similarity between the induced low molecular weight protein secretion by macrophages of 17 alpha-ethinylestradiol-treated and that of 2,3,7,8-tetrabromodibenzo-p-dioxin-treated mice was found.
Animals↗
A novel polypeptide p10 expressed in tumor-promoter-treated murine epidermis and in untreated neonatal murine epidermis.
12-O-Tetradecanoyl-phorbol-13-acetate (TPA) has been shown to induce a broad variety of morphological and metabolic changes in mouse skin. These include gross changes in cell growth as well as subtle changes regarding gene expression. Changes in the induction of specific proteins have been described, but it is not yet clear, if there exists a strict causal correlation between the induction of expression of specific proteins and the phenomenon of tumor promotion. At least some of these proteins might also be correlated with an accompanying hyperproliferative effect. This has been shown for protein p10, a 10 kDa protein not found in untreated adult mouse epidermis. The expression of this protein can be induced de novo after the topical administration of TPA. It is also shown, that p10 is expressed in untreated neonatal murine epidermis. Due to the induction of this protein by irritation, hyperproliferation-causing tumor promoters and due to the observable abundance in neonatal epidermis, this protein seems not strictly related to tumor promotion, but rather to proliferation and differentiation.
9,10-Dimethyl-1,2-benzanthracene↗
Homage to Takis S. Papas, 1935-1999.
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Alpharetrovirus↗
Antibody spectrum against the viral transactivator protein in patients with human immunodeficiency virus type 1 infection and Kaposi's sarcoma.
We analyzed patterns of antibody response to recombinant transactivator protein (human Immunodeficiency virus type 1 [HIV-1] tat) in serum samples from HIV-1-negative subjects (n = 60), HIV-1-infected asymptomatic patients (n = 20), HIV-1-infected patients with Kaposi's sarcoma (n = 25), and patients with Kaposi's sarcoma without HIV-1 infection. None of the healthy subjects possessed anti-tat immunoglobulin G (IgG) in their serum. All asymptomatic patients with HIV-1 infection were anti-tat IgG-positive. Epitope mapping revealed that these sera had anti-tat IgG to all the functional domains of tat protein. Histochemical studies on lymph nodes from five asymptomatic HIV-1-infected patients showed that, in all cases, tat-positive cells were present within the germinal center at the stage of follicular fragmentation containing immunoblasts and small lymphocytes. Of the 25 HIV-1-infected patients with Kaposi's sarcoma, 4 were anti-tat IgG-positive; however, the epitope analysis revealed that IgG to functional domains of tat protein--in particular to transactivating response element (TAR)-binding site--were absent. All patients with Kaposi's sarcoma without HIV-1 infection were anti-tat IgG-negative. Presence or absence of anti-tat IgG and a prevalence of different antibody profiles in different groups of patients indicated the pathophysiologic role of tat protein. Thus, a passive immunization with anti-tat IgG could be a useful strategy to influence the pathophysiologic state of the disease.
Antibodies, Viral↗
Expression and purification of recombinant HIV-1 tat protein using HIV-1-tat specific monoclonal antibodies.
We expressed tat protein from HIV-1 coding AA 1-67 (strain HIV-Bru) in the inducible vector pTrc 99A in E. coli. The tat coding region was cloned in the ATG site of the expression vector. A sequence coding for 15 AA was added at the 3'region as a molecular marker. After sonification, the tat protein was routinely detected in Western blots using the Mab developed by us. Following precipitation and centrifugation the resulting pellets were dissolved and purified by three different methods: 1. immunoaffinity-chromatography using Affi-gel HZ coupled with a Mab recognizing the N-terminal sequence of HIV-1-tat; 2. ion exchange chromatography using DEAE-52 cellulose, and 3. isoelectric focusing in free solution. The resulting protein extracts obtained from the three purification protocols were checked in ELISA with the antibody. The peak fraction from all the procedures showed tat activity. No cross reaction in the presence of sera from uninfected persons was observed. The results showed that the purification of tat protein using monoclonal antibodies leads to highly purified preparations.
Antibodies, Monoclonal↗
Inhibition of tat-mediated HIV-1-LTR transactivation and virus replication by sulfhydryl compounds with chelating properties.
D-Penicillamine, a structural analog of cysteine, has the ability to chelate metal ions and reacts with cysteine. We have shown earlier that D-Penicillamin is a potential inhibitor of tat-mediated transactivation of HIV-1-LTR (14) and possesses anti-HIV-1 activity (23). Following this approach, we evaluated the anti-tat and anti-HIV-1 activity of several sulfhydryl compounds with chelating properties. The tested compounds: N-(2-Mercapto-propionyl)-glycin (MPG), 2,3-Dimercapto-propanol (DMP) and 2,3-Dimercapto-propane-sulfonic acid (DMPS) exhibited an inhibitory effect on the tat-mediated transactivation in Jurkat cells, as well as in U937 cells. The highest inhibitory response was shown by DMP leading to about 50% inhibition of transactivation in Jurkat cels and an 80% inhibition in U937 cells. On the contrary, DMPS (30 micrograms/ml) had no inhibitory effect in U937 cells, but did exhibit a 50% inhibition of transactivation in Jurkat cells at 30 micrograms/ml. The antiviral activity of DMP and DMPS was evaluated in H9 cells. In the concentration range which is effective for antiviral effect, both the compounds were highly cytotoxic. Mercapto-propionyl-glycin, although a weak inhibitor of transactivation, was able to inhibit synctia formation to more than 90% and inhibit the viral antigene expression to about 70%. The concentration of MPG needed to achieve this antiviral effect was very high, but it had no cytotoxicity at this concentration. We suggest that a search for compounds using this approach may be useful in developing potential inhibitors of tat-mediated transactivation.
Anti-HIV Agents↗
Inhibition of histone-mediated gene transfer in eucaryotic cells by anti-histone IgG.
In our laboratory, the gene transfer efficiency of some lipofection reagents (lipofectine, lipofectamine, DOTAP and Dosper) and histones H3 and H4 was compared to that of DEAE-Dextran (64). The histones H3 and H4 were found to have the highest transfection efficiency of all the agents tested. In the present study we have analyzed other parameters important for gene delivery by the histones H3 and H4. We transferred the HIV-1 tat gene to Jurkat cells and measured the transactivation of HIV-1-LTR by the transactivator protein, expressed in Jurkat cells. The expression of CAT as a reporter gene hybridized to LTR was a direct measure of transactivation potential. In order to investigate whether the transfection was only due to the positive ionic character of the histones H3 and H4 we tested other histones (H1 and H2A) and polylysine in our system. Under our experimental conditions, neither polylysine, nor the histones H1 and H2A were able to promote gene transfer in Jurkat cells. The inability of these reagents to promote gene transfer was not dependent on DNA condensation; in EMSA (Electrophoretic Mobility Shift Assay) all these reagents exhibited a strong retardation of DNA. In the presence of anti-histone-IgG the transfection potential of histones H3 and H4 was diminished in a concentration - dependent manner. To investigate whether the histone antibodies inhibited the condensation of DNA by histones we carried out gel retardation assays (EMSA) in the absence and in the presence of histone antibodies. Anti-histone-IgG had no effect on the retardation of histone-DNA complexes; on the contrary, retardation was increased. This observation has led us to postulate two models for the possible mechanism by which the histones H3 and H4 catalyze gene transfer in eucaryotic cells.
Chloramphenicol O-Acetyltransferase↗
Cell transforming and oncogenic activity of 2,3,7,8--tetrachloro--and 2,3,7,8 tetrabromodibenzo-p-dioxin.
We have developed a host-mediated assay system for detection of the transforming activity of chemical carcinogens on peritoneal macrophages, directly, as well as indirectly acting carcinogenic substances administered intraperitoneally to NMRI mice could be examined in this way. Resident macrophages were recovered by peritoneal lavage from treated and untreated mice and cultured in soft agar. After 5-6 days normal and transformed cells could be distinguished. Statistical analysis comparing cells from 2, 3, 7, 8-tetrachlorodibenzo-dioxin (TCDD)-treated animals with those from control mice proved that the test is positive at least on a significance level of 5%, using the t-test. TCDD revealed a cell-transforming potential that showed a dose-dependent response in this host-mediated assay. The co-carcinogenic activity of TCDD was established in experiments with diphenylhydantoin. Low doses of diphenylhydantoin which did not exhibit any transforming potential in our system gained a high oncogenic potential by the simultaneous administration of low doses of TCDD, which also had no transforming activity. We have compared the cell transforming potential of TCDD with its bromo analog TBrDD. The cell transforming potential of TCDD is 7 times that of TBrDD. We have succeeded in establishing a permanent cell lined from mice treated with TBrDD. The oncogenicity of this cell line was tested in athymic nu/nu mice. Animals treated subcutaneously with these cells (1 x 10(6) cells) developed tumors at the injection site. Using monospecific antibodies to tumor necrosis factor alpha (TNF-alpha), we have found that TCDD stimulates the secretion of TNF-alpha. The experimental data reported here lead to the conclusion that TCDD has a carcinogenic as well as a co-carcinogenic activity and has the property to induce TNF-alpha.
Animals↗