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Biomedical subjects

J B Hansen

Publications and source records attributed to J B Hansen.

At least 127 records · Page 7Linked to original sources

9-acridinyl and 2-methoxy-6-chloro-9-acridinyl derivatives of aliphatic di-, tri-, and tetraamines. Chemistry, cytostatic activity, and schistosomicidal activity.

9-acridinyl derivatives of 1,6-hexanediamine, 1,8-octanediamine, bis(3-aminopropyl)amine, N,N'-bis(3-amino-propyl)piperazine, and N-ethyl-1,6-hexanediamine in the form of their hydrochlorides were prepared in high yields and converted into potential hetero bis DNA intercalating diacridines. The corresponding potential homo bis DNA intercalating reagents were prepared by heating the above amines with 9-chloroacridines. The chemical stability of the acridines was examined. Their cytostatic activity against Cloudman melanoma cells, in vitro, has been determined. The strongest cytostatic activity was observed for the acridine derivatives of the tri- and tetraamines. The schistosomicidal activity of selected acridine and diacridine derivatives against Schistosoma mansoni in mice was found to be insignificant. The S. mansoni egg development was apparently suppressed by this treatment.

Aminoacridines↗

Pseudomonas streptomycin resistance transposon associated with R-plasmid mobilization.

Plasmid pMG1 encodes resistance to gentamicin, streptomycin, sulfonamides, and mercuric ions and also mobilizes pRO161, a transfer-deficient plasmid derived from RP1. Upon mobilization, pRO161 acquires streptomycin resistance (Smr) and can subsequently be remobilized by pMG1 at significantly higher frequencies than pRO161 itself. Both the initial acquisition of Smr and the subsequent mobilization of the transfer-deficient plasmid are recA independent: thus, the Smr determinant appears to be located on a transposon, disignated Tn904. Tn904 transposes to a variety of other plasmids, including RP1, FP2, R388, K, pRO1600, and pBR322, and in some cases the acquisition of this transposon accompanied deletions in the target plasmid. When no deletion occurred, target plasmids gained 5.2 kilobase pairs of DNA and new restriction endonuclease cleavage sites for AvaI, BglII, PstI, SmaI, and SstI. Physical analysis of such plasmids showed that the Tn904 termini are inverted repeat DNA sequences of approximately 124 base pairs. After cloning into vector pRO1723, a single site for restriction endonuclease AvaI was identified within the Smr determinant of Tn904. In Escherichia coli, but not in Pseudomonas aeruginosa. Tn904 shows a gene dosage-dependent expression of streptomycin resistance.

DNA Transposable Elements↗

The development and application of a radioimmunoassay for rat phosphoenolpyruvate carboxykinase.

We employed a newly developed radioimmunoassay to quantitate P-enolpyruvate carboxykinase protein directly in liver and kidney of intact rats. The radioimmunoassay was dependent upon the presence of sodium dodecyl sulfate in the competitive binding reaction mixture. In conjunction with assayable activity measurements, the radioimmunoassay results also made it possible to assess the average specific activity of the enzyme. In the fed state, liver P-enolpyruvate carboxykinase was 0.89 microM (micromoles/kg of tissue) and the kidney enzyme was 1.90 microM. Following a 48-h fast, the enzyme in liver increased to 2.83 microM and that in kidney to 6.83 microM. Although liver enzyme concentration increased 3-fold, total liver enzymic activity was increased only about 30%. The discrepancy was due to the decrease in liver weight which occurred during fasting. The kidneys, which do not lose weight during fasting, had a 65% elevation of total organ activity. Total organ enzyme activity was predominantly dependent on changes in enzyme mass. In the fed state, the kidney/liver enzyme mass ratio was 0.44, in the fasted state it was 0.68. Variation in enzyme concentration, enzyme half-life, and response to tryptophan and chronic triamcinolone administration all gave evidence for organ-specific regulation.

Animals↗

Insulin and dexamethasone effects on liver fructose bisphosphatase.

Insuling perfusion of isolated diabetic rabbit liver and glucocorticoid perfusion of isolated normal rabbit liver resulted in an increased accumulation of immunoreactive fructose bisphosphatase (nmol/g). The insulin effect was maximal at 30 min with concentration (nmol/g) and specific activity (units/nmol) returning to normal within 2 h after insulin removal. Similarly, dexamethasone perfusion increased enzyme concentration and decreased its specific activity; however, the effects were maximal at 60-120 min. The results parallel those previously demonstrated in whole animals and establish that both hormones influence the enzyme by direct effects on the liver.

Animals↗

Streptococcus mutans genes that code for extracellular proteins in Escherichia coli K-12.

Chromosomal DNA from Streptococcus mutans 6715 (serotype g) was cloned into Escherichia coli K-12 by using the cosmid pJC74 cloning vector and a bacteriophage lambda in vitro packaging system. Rabbit antiserum against S. mutans extracellular proteins was used for immunological screening of the clone bank. Twenty-one clones produced weak to strong precipitin bands around the colonies, but only after the lambda c1857 prophage was induced by being heated to lyse the E. coli cells. None of the clones expressed enzyme activity for several known S. mutans extracellular enzymes. One of these clones contained a 45-kilobase recombinant plasmid designated pYA721. An 8.5-kilobase fragment of S. mutans DNA from pYA721 was isolated and recloned into the BamHI restriction site of the plasmid vector pACYC184 to construct pYA726. pYA726 contained all, or nearly all, of the gene for a surface protein antigen (the spaA protein) of S. mutans 6715. This was deduced from immunological studies in which extracts of cells harboring pYA726 reacted with antisera against both purified 6715 spaA protein (about 210,000 daltons) and the immunologically similar antigen I/II of serotype c strains of S. mutans. In addition, the S. mutans spaA protein was found to possess at least one antigenic determinant not present on the protein specified by pYA726. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of E. coli clone extracts revealed that pYA726 produced a polypeptide with a molecular mass of about 180,000 daltons which was predominantly found in the periplasmic space of E. coli cells. Antisera to the spaA protein of S. mutans reacted with extracellular protein from representative strains of S. mutans serotypes a, c, d, e, f, and g, but not b.

Antigens, Bacterial↗

Cloning and expression of the beta-D-phosphogalactoside galactohydrolase gene of Lactobacillus casei in Escherichia coli K-12.

Lactose metabolism in Lactobacillus casei 64H is associated with the presence of plasmid pLZ64. This plasmid determines both phosphoenolpyruvate-dependent phosphotransferase uptake of lactose and beta-D-phosphogalactoside galactohydrolase. A shotgun clone bank of chimeric plasmids containing restriction enzyme digest fragments of pLZ64 DNA was constructed in Escherichia coli K-12. One clone contained the gene coding for beta-D-phosphogalactoside galactohydrolase on a 7.9-kilobase PstI fragment cloned into the vector pBR322 in E. coli strain chi 1849. The beta-D-phosphogalactoside galactohydrolase enzyme isolated from E. coli showed no difference from that isolated from L. casei, and specific activity of beta-D-phosphogalactoside galactohydrolase was stimulated 1.8-fold in E. coli by growth in media containing beta-galactosides. A restriction map of the recombinant plasmid was compiled, and with that information, a series of subclones was constructed. From an analysis of the proteins produced by minicells prepared from transformant E. coli cells containing each of the recombinant subclone plasmids, it was found that the gene for the 56-kilodalton beta-D-phosphogalactoside galactohydrolase was transcribed from an L. casei-derived promoter. The gene for a second protein product (43 kilodaltons) was transcribed in the opposite direction, presumably under the control of a promoter in pBR322. The relationship of this second product to the lactose metabolism genes of L. casei is at present unknown.

Cloning, Molecular↗

Protective choledochoduodenostomy in multiple common duct stones in the aged.

Forty-nine patients more than 70 years of age had a follow-up examination two to eight years after choledochoduodenostomy for common duct stones or for common duct strictures following a previous biliary tract operation. Death followed operation in two patients and was not related to the choledochoduodenostomy itself. Results of the follow-up examination revealed that none of the patients had had signs or symptoms of cholangitis, jaundice or episodes of fever. One patient only had recurrent, but mild, biliary colic. Choledochoduodenostomy is well tolerated in old age and justified in elderly patients with multiple stones in dilated common ducts as a precaution against retained or recurrent common duct stones.

Age Factors↗

Dexamethasone effects on liver pyruvate kinase.

Dexamethasone in the medium perfusing isolated rabbit livers caused a fast-acting and reversible effect on liver pyruvate kinase. The effect was to lower the assayable V activity (units/g tissue) without changing the concentration (nmol/g enzyme protein). In effect, glucocorticoid lowered the specific activity (units/nmol of enzyme) by direct action on liver. The effect on liver pyruvate kinase is mediated by a relatively stable alteration; 30 min after perfusate (with steroid) was replaced by perfusate (without steroid), the effect remained strongly evident.

Animals↗

Characterization of the Streptococcus mutans plasmid pva318 cloned into Escherichia coli.

We further characterized the cryptic plasmid pVA318 of Streptococcus mutans. It had a contour length of 5.64 +/- 0.26 kilobases and a guanine-plus-cytosine content of 32 to 34 mol %. Upon cloning the pVA318 plasmid into the vector pBR322 in Escherichia coli, we made the following observations. The expression of tetracycline resistance by HindIII-cloned chimeras, where the insert was in the tetracycline resistance promotor, depended on the orientation of the pVA318 insert. Both HindIII-cloned chimeras segregated from polA(Ts) cells at a nonpermissive temperature. Chimeric molecules cloned with PstI initially showed much instability; the reason for this is unknown, although stable variants were obtained. Both HindIII-cloned variants and a PstI-cloned chimera produced a pVA318-specific protein of approximately 20,000 molecular weight in E. coli minicells. The biological function of this protein is not known; it had no bacteriocin activity against S. mutans or group A Streptococcus indicator strains, and it did not appear in the E. coli periplasm. We constructed a map of pVA318 for restriction endonucleases HindIII, HpaI, PstI, and HaeIII. A previously reported BamHI site in pVA318 did not appear in the pVA318 portion of any of our chimeric clones.

Cloning, Molecular↗

Intracellular concentration of skeletal and cardiac muscle phosphofructokinase in diabetic and normal animals.

An RIA has been developed for rabbit muscle PFK, an allosteric enzyme which catalyzes the pacemaker reaction of glucose utilization. The assay which could accurately measure as little as 12 ng (37.5 fmol) of the tetrameric enzyme was applied to extracts of skeletal and cardiac muscle of animals subjected to four different nutritional conditions and to alloxan diabetes. The concentration of enzyme remained unchanged in all conditions: mean value in anterior tibial muscle was 2.99 microM (S.D., 0.20; N, 32); in left ventricle, 0.66 microM (S.D., 0.07; N, 29). It may be concluded that control of the reaction is mainly, if not exclusively, by allosteric phenomena and not by insulin- or nutrition-dependent changes in intracellular concentration of the enzyme.

Animals↗

Analogues of [3H] chloramphenicol for photoaffinity labeling.

The synthesis of [3H]chloramphenicol and its erythro-diastereoisomer with specific activities of 1.25 Ci/mmol, and the further transformation of the [3H]chloramphenicol to a series of azido and diazo-substituted derivatives are described. The antibiotic activity of the compounds was considered insufficient for their use as photoaffinity labels.

Affinity Labels↗

The prophylaxis of septic complications in colo-rectal surgery. A controlled trial of metronidazole and oxytetracycline.

The prophylactic effect of systemic metronidazole on infectious complications following elective colo-rectal surgery was compared with the prophylactic effect of oxytetracycline. Allocation to receive metronidazole or oxytetracycline was random. A total of 137 patients were accepted for the trial, 67 receiving metronidazole and 70 oxytetracycline. The numbers of abdominal wound infections, intra-abdominal infections, fever and deaths were few in both groups, and the differences statistically insignificant. Clinically significant side effects of the antimicrobial agents were not observed. The influence of the antimicrobial agents on colonic flora was also studied. No significant difference in the effect on colonic anaerobic flora was demonstrated, whereas the aerobic flora was significantly suppressed by oxytetracycline. It is concluded that systemic metronidazole, although acting solely on the anaerobic flora, is as effective as oxytetracycline in preventing postoperative infectious complications following elective colo-rectal surgery. The prophylactic use of metronidazole is advocated.

Administration, Oral↗