Localization of the globin gene in the template active fraction of chromatin of Friend leukemia cells.

Friend leukemia cell chromatin has been fractionated into template active and inactive components. The globin gene sequence is associated with the template active component both prior to and after the cells are induced with dimethyl sulfoxide to synthesize hemoglobin and therefore appears to be in an active configuration in uninduced as well as in induced Friend leukemia cells. In cells which have lost the ability to produce hemoglobin, the globin gene sequence is not associated with the template active fraction of chromatin. These results demonstrate the success of the fractionation procedure.

Plant cell biodegradation of a xenobiotic nitrate ester, nitroglycerin.

The ability of plants to metabolize the xenobiotic nitrate ester, glycerol trinitrate (GTN, nitroglycerin), was examined using cultured plant cells and plant cell extracts. Intact cells rapidly degrade GTN with the initial formation of glycerol dinitrate (GDN) and the later formation of glycerol mononitrate (GMN). A material balance analysis of these intermediates indicates little, if any, formation of reduced, conjugated or cell-bound carbonaceous metabolites. Cell extracts were shown to be capable of degrading GTN with the simultaneous formation of GDN in stoichiometric amounts. The intermediates observed, and the timing of their appearance, are consistent with a sequential denitration pathway that has been reported for the microbial degradation of nitrate esters. The degradative activities of plant cells are only tenfold less than those reported for bacterial GTN degradation. These results suggests that plants may serve a direct degradative function for the phytoremediation of sites contaminated by organic nitrate esters.

Particle-associated RNA dependent DNA polymerase and high-molecular-weight RNA in a human cell line derived from polycythemia vera bone marrow.

A particle fraction with a density of 1.15-1.19 g/cm3 was isolated from the cytoplasm of a human cell line established in culture from the bone marrow of an untreated patient with polycythemia vera. Electron micrographs of cross sections of cells and cell homogenates revealed virus-like particles on which DNA could be synthesized. An RNA-dependent DNA polymerase, isolated from the particles, preferred poly(rA)-oligo(dT) over poly(dA)-oligo(dT) and was able to polymerize deoxyguanosine monophosphate in a reaction stimulated by poly(rC)-oligo(dG).

Molecular cloning and sequence of the gene for p9Ka. A cultured myoepithelial cell protein with strong homology to S-100, a calcium-binding protein.

The gene for p9Ka, a protein of molecular weight 9000 that is expressed in cultured rat mammary myoepithelial cells, has been isolated from a normal rat genomic library in bacteriophage lambda, by its ability to hybridize to a cloned complementary DNA corresponding to p9Ka mRNA. The cloned rat genomic DNA fragment hybridized to translatable p9Ka mRNA. A nucleotide sequence of 2340 base-pairs of genomic DNA surrounding the p9Ka cDNA sequence has been obtained; the gene contains one intervening sequence of 675 nucleotides. The 3' end of the p9Ka mRNA has been identified on the gene sequence to be 13 nucleotides downstream from a poly(A) addition signal AATAAA. The gene contains an open reading frame of 101 amino acid residues, which is the only open reading frame in the entire gene long enough to encode a protein of molecular weight at least 9000. This hypothetical protein sequence shows greater than 40% homology to rat or bovine S-100 protein and over 30% homology to bovine intestinal calcium-binding protein. The results suggest that p9Ka may be related to a family of low molecular weight calcium-binding proteins.

Effects of methadone on thyroid function in mother, fetus, and newborn.

The effects of maternal use of methadone during pregnancy on tests of thyroid function were studied in 14 women and their offspring. Twelve normal nonaddicted women and their infants served as a control group. Concentrations of thyrotropin (TSH), thyroxine (T4), and triiodothyronine (T3) were not significantly different in maternal and cord serum, and in infant serum at age 24 hours. There was no significant difference in infants' TSH concentrations at 2, 3, and 7 days of age. At 2 and 7 days, T3 levels were significantly higher in the methadone than in the control group (P = .01 and less than .01, respectively). At 3 days, differences were of borderline significance (P = .08). Increased T4 levels in the methadone group of infants were statistically significant at 2 days (P less than .01) and at 3 and 7 days (P = .05). The mechanisms responsible for the biochemical evidence of hyperthyroidism were not determined, but may have been related to altered autonomic function and/or increased metabolic activity which occurs during neonatal narcotic withdrawal.

Coamplification of 3-hydroxy-3-methylglutaryl coenzyme A reductase genes in methotrexate-resistant human leukemia cell lines.

Methotrexate (MTX)-resistant K562 human myelocytic leukemia sublines with 20- and 200-fold amplified dihydrofolate reductase (DHFR) genes localized to homogeneously staining regions (HSRs) on the long arms of chromosomes 5, 6, and 19 were used to examine whether other genes mapping close to the DHFR genes were coamplified. The gene for 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, located on chromosome 5q13.3-14, was coamplified 4-14-fold, corresponding to the levels of resistance exhibited by these cells. Similar observations were made with a MTX-resistant subline of the promyelocytic leukemia cell line, HL-60R, with 200 gene copies of DHFR. These observations indicate a tight linkage of DHFR and HMG-CoA genes on chromosome 5q.

Induction of protective immunity in chickens immunised with plasmid DNA encoding infectious bursal disease virus antigens.

Direct DNA inoculations were used to determine the efficacy of gene immunisation of chickens to elicit protective immune responses against infectious bursal disease virus (IBDV). The vp2 gene of IBDV strains GP40 and D78, and the vp2-vp4-vp3 encoding segment of strain D78 were cloned in an expression vector which consisted of human cytomegalovirus (HCMV) immediate early enhancer and promoter, adenovirus tripartite leader sequences and SV40 polyadenylation signal. For purification of vaccine-quality plasmid DNA from E. coli, an effective method was developed. Chickens were vaccinated by inoculation of DNA by two routes (intramuscular and intraperitoneal). Two weeks later, chickens were boosted with DNA, and at 2 weeks post-boost, they were challenged with virulent IBDV strain. Low to undetectable levels of IBDV-specific antibodies and no protection were observed with DNA encoding VP2. However, plasmids encoding VP2-VP4-VP3 induced IBDV-specific antibodies and protection in the chickens. DNA immunisation opens a new approach to the development of gene vaccines for chickens against infectious diseases.

Anaphylaxis to artesunate?

Artesunate, an artemissin derivative is a highly efficacious and relatively safe antimalarial agent. Common adverse reactions to artemissin derivatives are nausea, vomiting, anorexia and dizziness. More serious but less-frequent toxic effects of artesunate use are neutropenia, anemia, hemolysis, elevated liver enzymes and severe allergic reactions. However, anaphylactic reaction to artesunate is a rare entity. Here, we report a case of anaphylaxis to parenteral artesunate and its successful management in a female patient to whom intravenous artesunate was administered during surgery under general anesthesia.

Knowledge about Anaesthesia and Anaesthesiologist Amongst General Population in India.

SUMMARY: Anaesthesiologists are playing a decisive role in patient management. Present day anaesthesiology is based on the use of newer and safer drugs, better patient monitoring, pain management and critical care. But the general public knows little of these developments. The study was undertaken to assess the perception regarding the anaesthesiology and anaesthesiologists among the general population. The present study was conducted on 300 persons (patient, patient's attendant and medical undergraduates) between 18-75 years of age to assess the knowledge regarding anaesthesiology and the anaesthesiologists. All collected data were categorized into 5 groups as per the educational status of the study population. Perception of anaesthesiologist as a doctor in illiterate, graduate and postgraduate population was 19.51%, 58.57% and 87.88% respectively. Anaesthesiology as a separate medical discipline was not known to 100%, 73.87%, 64.29% and 51.52% of the illiterate, upto matriculation, graduate and postgraduate population respectively. Among the population who knew something about general anaesthesia, none from upto matriculation and 33.87%, 44.83% and 100% from the graduate, post graduate and medical undergraduate groups respectively knew that anaesthesia is administered with specialized equipments along with monitoring. Illiterates did not know about regional anaesthesia, while most of others had some knowledge about it. The results of the study reflect the wide spread ignorance and misconceptions about anaesthesiology and anaesthesiologists still prevalent in public in India.

Organization of soybean chalcone synthase gene clusters and characterization of a new member of the family.

Chalcone synthase (CHS; EC 2.3.1.74), the first committed enzyme of the multibranched pathway of flavonoid/isoflavonoid biosynthesis is encoded by a multigene family in soybean, (Glycine max L. Merrill). Our results suggest that this gene family comprises at least seven members, some of which are clustered. We have identified four chs clusters in the allo-tetraploid G. max genome and chs5, a newly characterized member of the chs gene family is present in two of them. We describe the complete nucleotide sequence of chs5, the identification of its immediate neighbors and the organization of the four hitherto identified chs clusters in the Gm genome.