The inhibitory receptor NKG2A determines lysis of vaccinia virus-infected autologous targets by NK cells

Signals transduced by inhibitory receptors that recognize self-MHC class I molecules prevent NK cells from being activated by autologous healthy target cells. In order for NK cells to be activated upon contact with an infected cell, the balance between the activating and inhibitory signals that regulate NK cell function must be altered in favor of activation. By studying liver-derived NK cells, we show that only a subpopulation of NK cells expressing high levels of the inhibitory receptor NKG2A are able to lyse autologous vaccinia-infected targets, and that this is due to selective down-regulation of HLA-E. These data demonstrate that release from an inhibitory receptor:ligand interaction is one mechanism that permits NK cell recognition of a virally infected target, and that the variegated expression of inhibitory receptors in humans generates a repertoire of NK cells with different antiviral potentials.     

RasGRP1 transmits prodifferentiation TCR signaling that is crucial for CD4 T cell development

TCR signaling plays a governing role in both the survival and differentiation of bipotent double-positive thymocytes into the CD4(+) and CD8(+) single-positive T cell lineages. A central mediator of this developmental program is the small GTPase Ras, emitting cytoplasmic signals through downstream MAPK pathways and eventually affecting gene expression. TCR signal transduction orchestrates the activation of Ras by integrating at least two Ras-guanyl nucleotide exchange factors, RasGRP1 and Sos. In this study, we have characterized the relationship between RasGRP1 function and its potential roles in promoting ERK activity, cell survival, maturation, and lineage commitment. Investigations on RasGRP1(-/-) mice expressing a transgenic (Tg) MHC class II-restricted TCR revealed that the development of CD4 T cells expressing this Tg TCR is completely dependent on RasGRP1. Unexpectedly, a small number of functional CD8 single-positive thymocytes expressing the Tg MHC class II-restricted TCR exists in mutant mice. In addition, RasGRP1(-/-) double-positive thymocytes exhibit marked deficits in TCR-stimulated up-regulation of the positive selection marker CD69 and the antiapoptotic protein Bcl-2, whereas CD5 induction is unaffected. To evaluate the role of RasGRP1 in providing cellular survival signaling, we enforced Bcl-2 expression in RasGRP1(-/-) thymocytes. These studies demonstrate that RasGRP1 function cannot be fully complemented by Tg Bcl-2 expression. Therefore, we propose that RasGRP1 transmits differentiation signaling critically required for CD4 T cell development.     

Salinity tolerance of mung bean (Vigna radiata L.): Seed production

Plant growth and seed yield of mung bean were studied in sand culture at different levels of NaCl [0, 50, 100, 150, 200, 250 mM] in the root medium. Results showed that both dry matter yield and seed yield of plants grown for 14 weeks at 50 mM NaCl and 100 mM NaCl were around 60 % and 25 %, respectively of those for plants grown in control solution. Higher concentrations caused wilting and necrosis of leaves. Very effective exclusion of Na and Cl from salt grown mung bean seed was observed with concommitant high accumulation of Na and Cl in the stem. It is speculated that mung bean plant stem may act as a ‘sink’ for NaCl during the reproductive stage of the plant growth cycle.     

Separation of motile human spermatozoa by means of a glass bead column

Fertility potential of semen depends upon the presence of viable and motile spermatozoa. A variety of techniques has been reported whereby motile spermatozoa are separated from semen for use in homologous insemination. Using a column of glass beads, we tested various sizes for spermatozoa-separating efficiency. This procedure is quick and simple and results in a signficantly better recovery of motile and viable spermatozoa from poor-quality semen. Therefore it has the potential for use in intrauterine insemination for treating male factor infertility.     

Relationships between the physiological characteristics and pathogenicity of Nocardia brasiliensis

The analysis of 19 physiological properties of 28 strains of Nocardia brasiliensis isolated from soil reveal differences which could be of great value in distinguishing pathogenic from nonpathogenic strains.     

The methylated nucleotide sequences in HELA cell ribosomal RNA and its precursors

The methylated nucleotide sequences in HeLa cell ribosomal RNA and its nucleolar precursors were examined by RNA fingerprinting and sequencing methods. 18 S RNA was found to contain approximately 46 methyl groups, 28 S RNA some 70 methyl groups and 5.8 S RNA one methyl group. Most methyl groups occur in different T₁ ribonuclease oligonucleotides, and most of these were recovered approximately once per molecule of 18 S or 28 S RNA. There are also, however, several multiply methylated oligonucleotides, a few short products that occur more than once and a few “fractional” products. The great majority of methylations occur at the level of 45 S RNA, but six further methylations occur late during the maturation of 18 S RNA, and one fractional one occurs during 28 S maturation. The transcribed spacer regions of the precursor molecules are unmethylated. Chemical analysis of the methylated components and sequences indicates that all except five “early” methylations are on ribose groups, the remaining five being on bases within the 28 S sequence. The late methylations are all on bases, four of those on 18 S RNA giving rise to the sequence, … Gpm₂^post6Apm₂^post6ApCp… The product, pCmpUp, previously reported by Choi &; Busch (1970) as being the 5′ end-group of rat hepatoma 28 S, 32 S and 45 S RNA, is not present in HeLa cell 28 S RNA or its precursors. Implications of this work are discussed.     

Adsorption of 5′-AMP and catalytic synthesis of 5′-ADP onto phosphate surfaces: Correlation to solid matrix structures

A non-enzymatic formation of 5-ADP starting from phosphorylation of 5-AMP in the presence of either calcium phosphate or calcium pyrophosphate precipitates is reported. This reaction is taken as a model for the study of heterogeneous catalysis of transphosphorylation in prebiotic conditions. Experiments were performed in completely aqueous media and in media containing dimethyl sulfoxide (Me₂S0), to simulate periods of dehydration in primitive aquatic environments. It has been observed that the nucleotide is adsorbed onto both calcium phosphate and calcium pyrophosphate in accordance with Langmuir isotherms. Adsorptive capacity and affinity of the precipitates for nucleotide are changed by the presence of Me₂SO, suggesting that the interaction between biomonomers and surfaces can be modulated by the degree of hydration of the anionic components of these compounds. In completely aqueous environments, formation of 5-ADP from 5-AMP adsorbed on precipitates of calcium phosphate and calcium pyrophosphate is very small. However, in the presence of 60% Me₂SO this synthesis increases by factors of 3 and 6 for surfaces of calcium phosphate and calcium pyrophosphate, respectively, and follows first-order kinetics. Determinations of free energy changes show that phosphorylation of 5-AMP adsorbed to these precipitates is thermodynamically favorable. Depending on the precipitation time of the samples and the composition of the medium, structural analysis of these precipitates by electron and X-ray diffraction shows changes in their cristallinity grade. It is proposed that these changes are responsible for the modulation of the quantity of adsorbed nucleotides to the surface of solid matrices as well as the catalytic activity of the precipitates.Abbreviations 5-AMP 5-adenosine monophosphate - 5-ADP 5-adenosine diphosphate - BTP l,3-bis[tris(hydroxymethyl)-methylamino]propane - CTEM conventional transmission electron microscopy - Tris tris(hydroxymethyl)aminomethane - Pi (H₂PO ₄ ^– /HPO ₄ ^2– ) orthophosphate - Pi.Ca calcium phosphate - PPi (H₃P₂O ₇ ^– /H₂P₂O ₇ ^2– ) pyrophosphate - PPi.Ca calcium pyrophosphate - EGTA [ethylenebis(oxyethylene)nitrilo]tetraacetic acid This work has been submitted to the Department of Biochemistry, Institute of Biomedical Sciences, UFRJ, by A.C.T. in partial fulfillment of requirements for the MS degree.     

Expression of the human beta-amyloid protein of Alzheimer's disease specifically in the brains of transgenic mice

The human beta-amyloid protein is deposited in senile plaques and in the cerebro-vasculature of people with Alzheimer's disease and Down's syndrome. The precise role of beta-amyloid in Alzheimer's disease pathology is presently unknown. To study the properties of beta-amyloid in vivo, we generated transgenic mice that harbor the gene for the carboxyl-terminal 100 amino acids of the human amyloid precursor protein, beginning with the beta-amyloid region, under control of the JC viral early region promoter. The mRNA is expressed exclusively in brain tissue. Further, we demonstrate increased levels of beta-amyloid immunoreactivity on fixed brain tissue. These animals will be useful as a model to study beta-amyloid deposition and its consequences.