The stability of L-ATC hydrolase participating in L-cysteine production

Summary In the production of L-cysteine from D,L-ATC stability of the relevant enzymes produced byPseudomonas sp. was tested, and strategies to improve the stability of L-ATC hydrolase were investigated in view of water activity and ionic strength. Among the three enzymes which participate in L-cysteine production, i.e., ATC racemase, L-ATC hydrolase, and S-carbamyl-L-cysteine hydrolase, L-ATC hydrolase is the least stable. Various mixtures of salts and sorbitol were added to adjust the water activities of the tested solutions. As water activity decreased from 0.93 to 0.80, the stability of L-ATC hydrolase was sharply enhanced. In the absence of sorbitol the stability of L-ATC hydrolase increased in proportion to ionic strength. Even though enzyme stability was not good at a low ionic strength, it was enhanced by lowering water activity with addition of sorbitol. The half life of L-ATC hydrolase in sorbitol-salt mixtures increased by tenfold to twentyfold compared to that of a control.     

Identification of three genetic loci controlling leaf senescence in Arabidopsis thaliana

Four mutants that show the delayed leaf senescence phenotype were isolated from Arabidopsis thaliana . Genetic analyses revealed that they are all monogenic recessive mutations and fall into three complementation groups, identifying three genetic loci controlling leaf senescence in Arabidopsis . Mutations in these loci cause delay in all senescence parameters examined, including chlorophyll content, photochemical efficiency of photosystem II, relative amount of the large subunit of Rubisco, and RNase and peroxidase activity. Delay of the senescence symptoms was observed during both age-dependent in planta senescence and dark-induced artificial senescence in all of the mutant plants. The results indicate that the three genes defined by the mutations are key genetic elements controlling functional leaf senescence and provide decisive genetic evidence that leaf senescence is a genetically programmed phenomenon controlled by several monogenic loci in Arabidopsis . The results further suggest that the three genes function at a common step of age-dependent and dark-induced senescence processes. It is further shown that one of the mutations is allelic to ein2-1 , an ethylene-insensitive mutation, confirming the role of ethylene signal transduction pathway in leaf senescence of Arabidopsis .     

Ginkgolide B production in cultured cells derived from Ginkgo biloba L. leaves

Callus cultures and cell suspension cultures derived from Ginkgo biloba L. leaves produced ginkgolidc B. In cell suspension cultures, the production reached a maximum by the 13th day of subculture and followed by a sharp decrease. The medium of Murashige and Skoog induced the highest ginkgolide B content in cultures while the medium of Schenk and Hildebrandt promoted cell growth. For the maximal production of ginkgolide B, cells were cultured in Murashige and Skoog medium modified to contain 1.0 mg/l of -naphthaleneacetic acid, 0.1 mg/1 of kinetin, 30 g/1 sucrose and 1.25 mM potassium phosphate with a molar ratio of ammonium to nitrate ions of 1 3.Abbreviations B5 Gamborg et al (1968) medium - GKB Ginkgolide B - MS Murashige and Skoog (1962) medium - NAA -naphthaleneacetic aicd - SH Schenk and Hildebrandt (1972) medium     

HhaI and HpaII DNA methyltransferases bind DNA mismatches, methylate uracil and block DNA repair.

The hydrolytic deamination of 5-methylcytosine (5-mC) to thymine (T) is believed to be responsible for the high mutability of the CpG dinucleotide in DNA. We have shown a possible alternate mechanism for mutagenesis at CpG in which HpaII DNA-(cytosine-5) methyltransferase (M.HpaII) can enzymatically deaminate cytosine (C) to uracil (U) in DNA [Shen, J.-C., Rideout, W.M., III and Jones, P.A., Cell, 71, 1073-1080, (1992)]. Both the hydrolytic deamination of 5-mC and enzymatic deamination of C create premutagenic DNA mismatches (G:U and G:T) with the guanine (G) originally paired to the normal C. Surprisingly, we found that DNA-(cytosine-5) methyltransferases have higher affinities for these DNA mismatches than for their normal G:C targets and are capable of transferring a methyl group to the 5-position of U, creating T at low efficiencies. This binding by methyltransferase to mismatches at the recognition site prevented repair of G:U mismatches by uracil DNA glycosylase in vitro.     

Effect of temperature on the growth,total lipid content and fatty acid composition of recently isolated tropical microalgae Isochrysis sp., Nitzschia closterium,Nitzschia paleacea,and commercial species Isochrysis sp. (clone T.ISO)

The effect of temperature from 10 °C to 35 °C on the growth, total lipid content, and fatty acid composition of three species of tropical marine microalgae, Isochrysis sp., Nitzschia closterium, N. paleacea (formerly frustulum), and the Tahitian Isochrysis sp. (T.ISO), was investigated.Cultures of N. closterium, Isochrysis sp. and T.ISO grew very slowly at 35 °C, while N. closterium did not grow at temperatures higher than 30 °C or lower than 20 °C. N. paleacea was low-temperature tolerant, with cells growing slowly at 10 °C. N. paleacea produced the highest percentage of lipids at 10 °C, while the other species produced maximum amounts of lipid at 20 °C. None of the species maintained high levels of polyunsaturated fatty acids (PUFAs) at high growth temperature and there was a significant inverse relationship between the percentage of PUFAs and temperature for N. paleacea. A curved relationship was found between temperature and percentage of PUFA for N. closterium and tropical Isochrysis sp., with the maximum production of PUFA at 25 °C and 20 °C, respectively. The two Nitzschia species produced higher levels of the essential fatty acid eicosapentaenoic acid [20:5(n-3)] at lower growth temperatures, but the two Isochrysis species had little change in percentage of 20:5(n-3) with temperature. Only T.ISO had the highest percentage of 22:6(n-3) at lowest growth temperature (11.4% total fatty acids at 10 °C).School of Mathematical and Physical SciencesAuthor for correspondence     

Vasoactive properties of urotensins I and II in a columbiform and three galliform birds, and a bioassay for urotensin I

Summary Crude extracts ofGillichthys urophyses and chromatographically purified urotensin I (UI) and urotensin II (UII) (fromCatostomus urophyses) were injected intravenously intoCoturnix coturnix japonica, Colinus virginianus, Alectoris graeca (Galliformes) andColumba livia (Columbiformes). Changes in arterial blood pressure were monitored. UI elicited dose-dependent vasodepressor responses in all birds. Thioglycollate treatment abolished the depressor action of arginine vasotocin but not that of UI (in all birds). UII was a pressor agent inCoturnix andColinus, both members of the Galliformes. However, the hormone had no pressor activity inColumba, a member of the Columbiformes, and in another member of the Galliformes,Alectoris. The pressor effect of UII was also dose-dependent. Injection of crude urophysial extract intoColinus andCoturnix, therefore, elicited biphasic responses. UII effects can be abolished by prior incubation with carboxypeptidase A. Intravenous injection of the -adrenoceptor blocker, phenoxybenzamine, prior to urotensin injections had no effect on the responses to urotensins. As the chukar,Alectoris graeca, is sensitive to UI and resistant to anesthesia and surgery, and does not readily develop tachyphylaxis to repeated UI injections, its use is recommended as a routine bioassay animal for UI.     

Chromosomal and histological changes in the reproductive organs of infertile men

Summary The chromosomal changes in the process of spermatogenesis in 27 infertile men have been examined. Normal chromosomal meiotic activity was found in 44% of cases, various chromosomal anomalies were seen in 18%, and no cells in meiosis were detected in 37% of cases.     

An in vitro comparison of Trypanosoma spp. (subgenus Schizotrypanum) from bats

Summary Epimastigotes of Trypanosoma vespertilionis from diphasic blood agar cultures were on the average longer and the distance between the nucleus and kinetoplast greater than epimastigotes of T. hedricki, T. myoti and T. dionisii. Also, no yellow granules were seen in the epimastigotes of T. vespertilionis whereas they were obvious in the other three species. Long thin trypomastigotes which are characteristic of T. hedricki, T. myoti and T. dionisii cultures were not seen in T. vespertilionis. T. dionisii was much less infective to fibroblasts from mice and did not develop in fibroblasts from chicken, as did T. hedricki and T. myoti. Blood trypomastigotes were seen in chicken embryos inoculated with blood agar cultures of T. hedricki and T. myoti, but none was seen in embryos infected with T. dionisii.The cultural characteristics examined could not be used to differentiate T. hedricki from T. myoti. ac]19810317