Regulation of urea-uptake in the cyanobacterium Anabaena doliolum

Abstract The utilization of urea was studied in the cyanobacterium Anabaena doliolum . The uptake of urea was unaltered in the presence of ammonium. The cells receiving ATP exogenously showed an induced level of urea-uptake as compared with the control cells. Urease inhibitor acetohydroxamic acid and hydroxyurea as well as glutamate analogue, MSO, did not affect the uptake of urea. These results suggest: (1) urea and ammonia have different uptake sites, (2) urea-uptake is an energy dependent process, and (3) during short-term experiments, urea uptake is not linked with the enzyme urease or the ammonium assimilating enzyme glutamine synthetase.     

UDP-Gal: <Emphasis Type="Italic">N</Emphasis>-acetylglucosamine β 1–4 galactosyltransferase expressing live attenuated parasites as vaccine for visceral leishmaniasis

As compared to cutaneous leishmaniasis, vaccination against visceral leishmaniasis (VL) has received limited attention. In this study, we demonstrate for the first time that an UDP-Galactose: N-acetylglucosamine β 1–4 galactosyltransferase (GenBank Accession No. EF159943) expressing attenuated LD clonal population (A-LD) is able to confer protection against the experimental challenge with the virulent LD AG83 parasite. A-LD was also effective in established leishmania infection. The vaccinated animals showed both cell mediated (in vitro T-cell proliferation, and DTH response) and humoral responses (Th1 type). These results demonstrate the potential of the attenuated clones as an immunotherapeutic and immunoprophylactic agent against visceral leishmaniasis.     

Effect of vitamin B6 deficiency on pancreatic acpinar cell function

The present study was designed to determine the effect of vitamin B₆ deficiency on pancreatic acinar cell function. Rats were either fed $\text{ad}$$\text{lib}$ or rendered B₆-deficient by a purified B₆-deficient diet; half of the latter being replenished with IP pyridoxine before sacrifice. Body weight, pancreatic weight, RNA and DNA content were decreased in B₆-deficient animals. These changes were considered to be due to inanition resulting from decreased food intake. Amylase content of pancreas in B₆-deficient animals was less compared with B₆-replenished animals. Although slightly higher in B₆-deficient animals, the incorporation of L-phenylalanine¹⁴C into total tissue proteins was not significantly different in the three groups of animals. On B₆-replenishment, incorporation of L-phenylalanine¹⁴C into nascent proteins was diminished in spite of higher tissue amylase and protein content. Vitamin B₆ deficiency decreased total RNA content and adenine-8-¹⁴C incorporation into RNA. DNA content was diminished but incorporation of thymidine-2-¹⁴C into DNA was increased. On replenishment with B₆, thymidine-2-¹⁴C incorporation decreased significantly compared to control animals. Secretion of amylase was diminished commensurate with decreased content. It is concluded from these studies that B₆-deficiency induced DNA injury, decreased RNA turnover and increased protein turnover resulting in diminished amylase content. These data indicate that B₆-deficiency so frequently encountered in alcoholism may contribute to the pancreatic injury in this clinical condition.     

Protoplast formation and regeneration in <Emphasis Type="Italic">Lactobacillus delbrueckii</Emphasis>

Method for production and regeneration of Lactobacillus delbrueckii protoplasts are described. The protoplasts were obtained by treatment with a mixture of lysozyme and mutanolysin in protoplast buffer at pH 6.5 with different osmotic stabilizers. The protoplasts were regenerated on deMan, Rogosa and Sharpe (MRS) with various osmotic stabilizers. Maximum protoplast formation was obtained in protoplast buffer with sucrose as an osmotic stabilizer using a combination of lysozyme (1 mg/ml) and mutanolysin (10 μg/ml). Maximum protoplast regeneration was obtained on MRS medium with sucrose (0.5 M) as an osmotic stabilizer. The regeneration medium was also applicable to other species of lactobacilli as well. This is, to our knowledge, the first report on protoplast formation and efficient regeneration in case of L. delbrueckii.     

Nif-hybrids of Enterobacter: selection for nif gene integration with chlorate

The nif gene group from Klebsiella can be transferred into Enterobacter cloacae by conjugation using Escherichia coli donor cells carrying the composite self-transmissible nif-plasmid pRD1. A small fraction of the hybrids obtained is stable upon prolonged passaging without selection. Their stability is due to integration of pRD1 into the chromosome. Such integration hybrids were chlorate resistant, and nitrate reductase negative, which indicated that integration preferentially occurred within one of the genes for the production or functioning of this enzyme. Chlorate resistance could, therefore, be used to select for additional nitrate reductase-negative sublines with pRD1 in their chromosome. Such sublines have been analyzed further for the presence of nif genes, other pRD1 markers, and for stability. In all except one the complete plasmid seems to have been integrated. Some tend to revert to nitrate utilisation (chlorate sensitivity).