Non-T cell activation linker (NTAL): a transmembrane adaptor protein involved in immunoreceptor signaling

A key molecule necessary for activation of T lymphocytes through their antigen-specific T cell receptor (TCR) is the transmembrane adaptor protein LAT (linker for activation of T cells). Upon TCR engagement, LAT becomes rapidly tyrosine phosphorylated and then serves as a scaffold organizing a multicomponent complex that is indispensable for induction of further downstream steps of the signaling cascade. Here we describe the identification and preliminary characterization of a novel transmembrane adaptor protein that is structurally and evolutionarily related to LAT and is expressed in B lymphocytes, natural killer (NK) cells, monocytes, and mast cells but not in resting T lymphocytes. This novel transmembrane adaptor protein, termed NTAL (non-T cell activation linker) is the product of a previously identified WBSCR5 gene of so far unknown function. NTAL becomes rapidly tyrosine-phosphorylated upon cross-linking of the B cell receptor (BCR) or of high-affinity Fcgamma- and Fc epsilon -receptors of myeloid cells and then associates with the cytoplasmic signaling molecules Grb2, Sos1, Gab1, and c-Cbl. NTAL expressed in the LAT-deficient T cell line J.CaM2.5 becomes tyrosine phosphorylated and rescues activation of Erk1/2 and minimal transient elevation of cytoplasmic calcium level upon TCR/CD3 cross-linking. Thus, NTAL appears to be a structural and possibly also functional homologue of LAT in non-T cells.     

Primary hyperparathyroidism associated with hypocalcemia in a patient presenting with kidney disease

Elevated serum parathyroid hormone (PTH) level together with hypocalcemia in chronic kidney disease usually suggests secondary hyperparathyroidism. However, primary hyperparathyroidism should also be considered, especially if concomitant vitamin D deficiency is suspected. We report a case of parathyroid adenoma associated with hypocalcemia and metabolic bone disease in a patient presenting with kidney disorder. The patient was successfully treated by parathyroidectomy that was preceded and followed by intensive calcium and vitamin D supplementation.     

Development and characterization of anionic liposaccharides for enhanced oral drug delivery

The aim of this study was to synthesize charged amphoteric molecules, which after complexation with poorly bioavailable drugs would have the potential to improve their oral uptake. Novel anionic liposaccharide derivatives containing d-glucose and lipoamino acids were synthesized by solution phase peptide synthesis. High sensitivity isothermal titration microcalorimetry was used to determine the critical aggregation concentration and the thermodynamic profiles. Hemolytic and cytotoxic activities of the liposaccharides were studied and they revealed that the liposaccharides were non-toxic at the concentration used for oral administration. Mixing a model drug, tobramycin, with the liposaccharide containing two lipids formed aggregates around 200 nm, which increased tobramycin partitioning between n-octanol/water. The results suggested that the studied liposaccharide with two lipids was safe to apply biologically and may have an absorption enhancing activity on hydrophilic, orally poorly available drugs.     

Is rat liver affected by non-alcoholic steatosis more susceptible to the acute toxic effect of thioacetamide?

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic condition of the liver in the western world. There is only little evidence about altered sensitivity of steatotic liver to acute toxic injury. The aim of this project was to test whether hepatic steatosis sensitizes rat liver to acute toxic injury induced by thioacetamide (TAA). Male Sprague-Dawley rats were fed ad libitum a standard pelleted diet (ST-1, 10% energy fat) and high-fat gelled diet (HFGD, 71% energy fat) for 6 weeks and then TAA was applied intraperitoneally in one dose of 100 mg/kg. Animals were sacrificed in 24-, 48- and 72-h interval after TAA administration. We assessed the serum biochemistry, the hepatic reduced glutathione, thiobarbituric acid reactive substances, cytokine concentration, the respiration of isolated liver mitochondria and histopathological samples (H+E, Sudan III, bromodeoxyuridine [BrdU] incorporation). Activities of alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase and concentration of serum bilirubin were significantly higher in HFGD groups after application of TAA, compared to ST-1. There were no differences in activities of respiratory complexes I and II. Serum tumour necrosis factor alpha at 24 and 48 h, liver tissue interleukin-6 at 72 h and transforming growth factor β1 at 24 and 48 h were elevated in TAA-administrated rats fed with HFGD, but not ST-1. TAA-induced centrilobular necrosis and subsequent regenerative response of the liver were higher in HFGD-fed rats in comparison with ST-1. Liver affected by NAFLD, compared to non-steatotic liver, is more sensitive to toxic effect of TAA.     

Integration of genetic and physical maps of the chickpea (Cicer arietinum L.) genome using flow-sorted chromosomes

Cultivated chickpea is the third most important legume after field bean and garden pea worldwide. Despite considerable breeding towards improved yield and resistance to biotic and abiotic stresses, the production of chickpea remained stagnant, but molecular tools are expected to increase the impact of current improvement programs. As a first step towards this goal, various genetic linkage maps have been established and markers linked to resistance genes been identified. However, until now, only one linkage group (LG) has been assigned to a specific chromosome. In the present work, mitotic chromosomes were sorted using flow cytometry and used as template for PCR with primers designed for genomic regions flanking microsatellites. These primers amplify sequence-tagged microsatellite site markers. This approach confirmed the assignment of LG8 to the smallest chromosome H. For the first time, LG5 was linked to the largest chromosome A, LG4 to a medium-sized chromosome E, while LG3 was anchored to the second largest chromosome B. Chromosomes C and D could not be flow-sorted separately and were jointly associated to LG6 and LG7. By the same token, chromosomes F and G were anchored to LG1 and LG2. To establish a set of preferably diagnostic cytogenetic markers, the genomic distribution of various probes was verified using FISH. Moreover, a partial genomic bacterial artificial chromosome (BAC) library was constructed and putative single/low-copy BAC clones were mapped cytogenetically. As a result, two clones were identified localizing specifically to chromosomes E and H, for which no cytogenetic markers were yet available.     

Czech socio-economic deprivation index and its correlation with mortality data

Objectives:  The aim of the study was to create an index of socio-economic deprivation (SESDI) and to analyse correlation between SESDI and mortality data. Methods:  The SESDI components were selected from the census data (2001) at enumeration district and district level. Two methods were used for creating the SESDI: 1/ a sum of Z-scores of specific components (INDEX1); and 2/ standardized score – average values of specific components were divided by a maximum value of the specific component at the corresponding geographical level (INDEX2). Pearson’s correlation coefficient was used for assessing the relationship between indices, and between indices and mortality data (SMR). Results:  The final indices were applied to districts in the Czech Republic (N = 77). The correlation of INDEX1 and INDEX2 was high (r = 0.99). Analysis of relationships between degree of deprivation and total and selected specific SMR in the Czech Republic confirmed that mortality was associated with degree of deprivation. Conclusion:  The use of socio-economic deprivation indices in analysis of routinely collected mortality data in public health might help to explain health inequalities. Submitted: 14 November 2007; revised: 06 May 2008, 12 June 2008; accepted: 17 September 2008     

Characterization of seven new monoclonal antibodies against human DR, DR + DP and DQ1 + DQ3 antigens

Seven murine monoclonal antibodies were prepared that react with human class II antigens. Four of them (HL-39, MEM-12, MEM-24G, and MEM-32B: react with a monomorphic determinant dependent on association of heavy and light chains of DR antigens, two others (HL-38 and HL-40) recognize a monomorphic determinant localized on the light chains of DR and DP antigens. The antibody HL-37 is directed against a determinant present on DQ1 and DQ3, but not DQ2 molecules; at least in the case of DQ1, the epitope recognized is located on the light chain.     

Endogenous development, pathogenicity and host specificity of Eimeria cahirinensis Couch, Blaustein, Duszynski, Shenbrot and Nevo, 1997 (Apicomplexa: Eimeriidae) from Acomys dimidiatus (Cretzschmar 1826) (Rodentia: Muridae) from the Near East

Eimeria cahirinensis Couch et al. 1997 was found in faecal samples of Acomys dimidiatus from three different localities in the Near East. Twenty-two of 104 (21%) A. dimidiatus trapped on both the south- and north-facing slopes of “Evolution Canyon”, Lower Nahal Oren, Mt. Carmel, Israel in August 2001 and 2002 were infected with E. cahirinensis. Oocysts were also obtained from a single individual of A. dimidiatus trapped in Wadi Ramm, Jordan in the summer of 1999. Laboratory-reared spiny mice (Acomys spp.) were inoculated to determine the prepatent and patent period, sporulation time, site of infection, immunogenicity, pathogenicity, pathology and morphology of endogenous stages of E. cahirinensis. Both asexual and sexual stages were localised in the apical part of duodenal and jejunal villi. An experimental inoculation of representatives of several rodent genera revealed the host range of E. cahirinensis to be limited to the genus Acomys.