Synthesis of a cytosine nucleoside of 2-amino-2-deoxy-beta-D-xylofuranose.

1-(2-Amino-2-deoxy-beta-D-xylofuranosyl)cytosine (13) was synthesized by three routes: (a) coupling of 2-deoxy-3,5-di-O-p-nitrobenzoyl-2-(trifluoroacetamido)-D-xylofuranosyl chloride (5) with 2,4-dimethoxypyrimidine and subsequent treatment with methanolic ammonia, (b) coupling of 5 with 4-N-acetyl-2-O,4-N-bis(trimethylsilyl)cytosine followed by treatment with methanolic ammonia, and (c) thiation of 1-[3,5-di-O-acetyl-2-deoxy-2-(trifluoroacetamido)-beta-D-xylofuranosyl]uracil (6) by treatment with phosphorus pentasulfide in pyridine followed by amination of the resulting 4-thionucleoside 12 with metanolic ammonia. The best yield was obtained via route (a).     

Appearance of lymphocyte surface markers and functional responses in neonatal and young rabbit spleens

We examined spleen cells from newborn to 1-month-old rabbits for easily detectable surface immunoglobulin, complement receptors, and for in vitro proliferative responsiveness to anti-immunoglobulin antisera and several mitogens. From birth through the first month of life about 15% of the cells from rabbit spleens had easily detectable surface immunoglobulin while about 45% had C3 receptors. In adults as many as 77% of the spleen cells had easily detectable surface Ig but the proportion with C3 receptors remained about 45%. The proliferative response to anti-allotype antisera was present at birth, and was at adult levels by 1 month of age. The proliferative response to pokeweed mitogen was low when cells were obtained during the first week of life but was comparable in magnitude to the response of adult cells by 2 weeks of age. In vitro responsiveness to concanavalin A was present at low levels at birth and increased sharply during the first week. We did not observe significant stimulation of spleen cells from neonatal to 4-week-old rabbits by lipopolysaccharide from Salmonella typhosa. Our data suggest that lymphocyte surface markers and functional responses appear asynchronously in spleen cells of developing rabbits.     

Studies on plasma membranes. XXIII. Hormone-like action of concanavalin A on liver plasma membranes: inhibition of (Na+-K+)ATPase.

Concanavalin A inhibits the (Na+-K+)-ATPase activity of isolated rat-liver plasma membranes, while leaving the Mg2+-ATPase unaffected. Glucagon and cyclic AMP act supplementary to the lectin in the inhibition. The lectin effect is counteracted by insulin and L-epinephrine, and is completely abolished by the beta-adrenergic blocking agent propranolol. Results are discussed on the basis of the known interactions of concanavalin A with plasma membrane components, including its hormone-like action.     

A new method for determination of elastolytic activity using (14C) labeled elastin and its application to leukocytic elastase.

A new, highly sensitive and specific assay for elastolytic activity is described which employs insoluble elastin randomly labeled with [¹⁴C]. The substrate was prepared by labeling amino groups of the protein in vitro with [¹⁴C] methyl groups by reductive alkylation. The substrate was used to quantitate elastolytic activity from human leukocytes and to compare leukocytic elastase with pancreatic elastase. Purified human leukocytic elastase was approximately one-fourth as active as pancreatic elastase. Similar difference between leukocytic elastase and pancreatic elastase activities was found when the enzymes were tested against succinyl-L-alanyl-L-alanyl-L-alanine-p-nitroanilide, but not when t-BOC-L-alanine-p-nitrophenyl ester was used.     

The utilization of bromodeoxyuridine incorporation into DNA for the analysis of cellular kinetics

Recently developed differential staining techniques based on the incorporation of bromodeoxyuridine (BUdR) into DNA permits the unequivocal identification of metaphase cells which have replicated once, twice, and three or more times. This technique has the potential of being utilized in the examination of kinetics of dividing cell populations. This potential is examined in a phytohemagglutinin-stimulated lymphocyte system. Determinations of the effect of increasing concentrations of BUdR on the distribution of metaphase cells between different generation cycles reveals no inhibition of cellular kinetics below 35 μM. The ability to distinguish third generation metaphase cells from subsequent generations is examined through the determination of “labelled” centromeric regions. The applicability of this system to current cellular kinetics is discussed.     

Anti-salmonella properties of kefir yeast isolates: An in vitro screening for potential infection control

The rise of antibiotic resistance has increased the need for alternative ways of preventing and treating enteropathogenic bacterial infection. Various probiotic bacteria have been used in animal and human. However, Saccharomyces boulardii is the only yeast currently used in humans as probiotic. There is scarce research conducted on yeast species commonly found in kefir despite its claimed potential preventative and curative effects. This work focused on adhesion properties, and antibacterial metabolites produced by Kluyveromyces lactis and Saccharomyces unisporus isolated from traditional kefir grains compared to Saccharomyces boulardii strains. Adhesion and sedimentation assay, slide agglutination, microscopy and turbidimetry assay were used to analyze adhesion of Salmonella Arizonae and Salmonella Typhimurium onto yeast cells. Salmonella growth inhibition due to the antimicrobial metabolites produced by yeasts in killer toxin medium was analyzed by slab on the lawn, turbidimetry, tube dilution and solid agar plating assays. Alcohol and antimicrobial proteins production by yeasts in killer toxin medium were analyzed using gas chromatography and shotgun proteomics, respectively. Salmonella adhered onto viable and non-viable yeast isolates cell wall. Adhesion was visualized using scanning electron microscope. Yeasts-fermented killer toxin medium showed Salmonella growth inhibition. The highest alcohol concentration detected was 1.55%, and proteins with known antimicrobial properties including cathelicidin, xanthine dehydrogenase, mucin-1, lactadherin, lactoperoxidase, serum amyloid A protein and lactotransferrin were detected in yeasts fermented killer medium. These proteins are suggested to be responsible for the observed growth inhibition effect of yeasts-fermented killer toxin medium. Kluyveromyces lactis and Saccharomyces unisporus have anti-salmonella effect comparable to Saccharomyces boulardii strains, and therefore have potential to control Salmonella infection.     

基于“心与小肠相表里”理论探讨肠道微生态与骨质疏松症的关系

近年来相关研究显示,肠道微生态在骨质疏松症的发生发展中起着重要作用。中医脏腑理论密切关注脏腑之间的生理病理关系,以中医经典《内经》“心与小肠相表里”理论为基础,探讨心、小肠、肠道微生态与骨质疏松症之间的关系。研究发现肠道微生态可能是心系疾病导致骨质疏松症的途径之一,这一发现可能为骨质疏松症的研究与防治提供一定的理论依据。     

Some aquatic oligochaetes of the Nilgiris,South India

Eight species of aquatic oligochaetes, belonging to the families Naididae and Tubificidae, are reported, of which six species are new to the Nigiris. Tubifex tubifex known from Coonoor is reported again from this area.     

Butanedione treatment reduces receptor binding of a lysosomal enzyme to cells and membranes

Treatment of the lysosomal enzyme, α-L-iduronidase, with 2,3 butanedione, an arginine modifying reagent, under conditions where enzyme activity was unaffected, reduced by 50% the internalization of the enzyme into cultured human fibroblasts. The lowered rate of internalization was a result of a reduced binding of the enzyme to cell surface receptors. The butanedione treatment of α-L-iduronidase caused a 90% reduction of binding when isolated fibroblast membranes were used as the source of receptor. This marked reduction of binding was also seen when membranes from a rat chondrosarcoma were examined. Although there is ample evidence that the receptor recognizes mannose 6-phosphate residues on the enzyme, the results suggest that other structural features, such as arginine moieties, may also be important in iduronidase binding.