Transforming growth factor-ß1 functional polymorphisms in myeloablative sibling hematopoietic stem cell transplantation.

Hematopoietic stem cell transplantation (HSCT) with sibling donors (s.d.) is a life-saving intervention for patients with hematological malignancies. Numerous genetic factors have a role in transplant outcome. Several functional polymorphisms have been identified in TGF-ß1 gene, such as single-nucleotide polymorphism (SNP) at +29C>T within exon 1. Two hundred and forty five patient/donor pairs who underwent a s.d. HSCT in our centers were genotyped for this SNP. In the myeloablative cohort, +29CC donors were associated with an increase in severe chronic GvHD (32% vs 16%, hazard ratio (HR) 9.0, P=0.02). Regarding survival outcomes, +29CC patients developed higher non relapse mortality (NRM) (1-5 years CC 28-32% vs TC/TT 7-10%; HR 5.1, P=0.01). Recipients of +29TT donors experienced a higher relapse rate (1-5 years TT 37-51% vs TC 19-25% vs CC 13%-19%; HR 2.4, P=0.01) with a decreased overall survival (OS) (1-5 years TT 69-50% vs TC/CC 77-69%; HR 1.9, P=0.05). Similar to previous myeloablative unrelated donors HSCT results, we confirmed that +29CC patients had higher NRM. In addition we found that +29TT donors might be associated with a higher relapse rate and lower OS. These results should be confirmed in larger series. Identification of these SNPs will allow personalizing transplant conditioning and immunosuppressant regimens, as well as assisting in the choice of the most appropriate donor.

Evaluation of alternative reporter genes for the yeast two-hybrid system

The yeast two-hybrid system is a powerful tool for screening protein-protein interactions and has also been used for large-scale studies. We evaluated two protein-coding sequences as reporter genes for the yeast two-hybrid system, to determine if it was suitable as an alternative screening strategy. Aspergillus awamori glucoamylase activity results in clear haloes around colonies producing this enzyme after growth on starch plates and staining with iodine vapors. However, transcription activation by Gal4 on Gal-regulated promoters was insufficient for this type of phenotypic visualization. A modified gene of Aequoria victoria enhanced green fluorescent protein (EGFP) was tested to determine its suitability for interaction screenings with flow cytometry. When the EGFP reporter gene system was incorporated into the cells, Gal4 transcriptional activation produced sufficient fluorescence for detection with the flow cytometer, especially when there were strong interactions

[Absence of HTLV-I/II virus circulation in blood donors from the provinces of Santa Fe and Santiago del Estero]. / Ausencia de circulación de los virus HTLV-I/II en donantes de sangre de las provincias de Santa Fe y Santiago del Estero.

In the present study, we evaluated the seroprevalence of HTLV-I/II infection among the blood donors in Santa Fe and Santiago del Estero provinces. A total of 1327 serum samples from blood donors from Rafaela blood bank of Santa Fe province and 3382 serum samples from blood donors from Dr Edgar Bouzon blood bank of Santiago del Estero province were studied. The antibody screening was done by particle agglutination assay (PA) (SERODIA, Fujirebio Inc., Tokyo, Japan) or by enzyme immuno assay (EIA) (Abbott HTLV-I/HTLV-II EIA, Abbott, Germany). The "in house" indirect immunofluorescence assay (IFA) and Western blot (Bioblot HTLV Biokit, Barcelona, Spain) were used as confirmatory assays. All the samples resulted negative for specific antibodies against HTLV-I/II. These results suggest that HTLV-I/II are not circulating in low risk populations in these provinces or that the prevalences of infection would be lower than that reported by blood banks in other provinces of Argentina.

Ausencia de circulación de los virus HTLV-I/II en donantes de sangre de las provincias de Santa Fe y Santiago del Estero / Absence of HTLV-I/II virus circulation in blood donors from the provinces of Santa Fe and Santiago del Estero

In the present study, we evaluated the seroprevalence of HTLV-I/II infection among the blood donors in Santa Fe and Santiago del Estero provinces. A total of 1327 serum samples from blood donors from Rafaela blood bank of Santa Fe province and 3382 serum samples from blood donors from Dr Edgar Bouzon blood bank of Santiago del Estero province were studied. The antibody screening was done by particle agglutination assay (PA) (SERODIA, Fujirebio Inc., Tokyo, Japan) or by enzyme immuno assay (EIA) (Abbott HTLV-I/HTLV-II EIA, Abbott, Germany). The "in house" indirect immunofluorescence assay (IFA) and Western blot (Bioblot HTLV Biokit, Barcelona, Spain) were used as confirmatory assays. All the samples resulted negative for specific antibodies against HTLV-I/II. These results suggest that HTLV-I/II are not circulating in low risk populations in these provinces or that the prevalences of infection would be lower than that reported by blood banks in other provinces of Argentina.

Ausencia de circulación de los virus HTLV-I/II en donantes de sangre de las provincias de Santa Fe y Santiago del Estero / Absence of HTLV-I/II virus circulation in blood donors from the provinces of Santa Fe and Santiago del Estero

In the present study, we evaluated the seroprevalence of HTLV-I/II infection among the blood donors in Santa Fe and Santiago del Estero provinces. A total of 1327 serum samples from blood donors from Rafaela blood bank of Santa Fe province and 3382 serum samples from blood donors from Dr Edgar Bouzon blood bank of Santiago del Estero province were studied. The antibody screening was done by particle agglutination assay (PA) (SERODIA, Fujirebio Inc., Tokyo, Japan) or by enzyme immuno assay (EIA) (Abbott HTLV-I/HTLV-II EIA, Abbott, Germany). The "in house" indirect immunofluorescence assay (IFA) and Western blot (Bioblot HTLV Biokit, Barcelona, Spain) were used as confirmatory assays. All the samples resulted negative for specific antibodies against HTLV-I/II. These results suggest that HTLV-I/II are not circulating in low risk populations in these provinces or that the prevalences of infection would be lower than that reported by blood banks in other provinces of Argentina.(AU)

Ausencia de circulación de los virus HTLV-I/II en donantes de sangre de las provincias de Santa Fe y Santiago del Estero. / [Absence of HTLV-I/II virus circulation in blood donors from the provinces of Santa Fe and Santiago del Estero]

In the present study, we evaluated the seroprevalence of HTLV-I/II infection among the blood donors in Santa Fe and Santiago del Estero provinces. A total of 1327 serum samples from blood donors from Rafaela blood bank of Santa Fe province and 3382 serum samples from blood donors from Dr Edgar Bouzon blood bank of Santiago del Estero province were studied. The antibody screening was done by particle agglutination assay (PA) (SERODIA, Fujirebio Inc., Tokyo, Japan) or by enzyme immuno assay (EIA) (Abbott HTLV-I/HTLV-II EIA, Abbott, Germany). The [quot ]in house[quot ] indirect immunofluorescence assay (IFA) and Western blot (Bioblot HTLV Biokit, Barcelona, Spain) were used as confirmatory assays. All the samples resulted negative for specific antibodies against HTLV-I/II. These results suggest that HTLV-I/II are not circulating in low risk populations in these provinces or that the prevalences of infection would be lower than that reported by blood banks in other provinces of Argentina.

Perception of sweetness and bitterness in different vehicles.

In the present study, we investigated taste-taste, taste-vehicle, and simultaneous taste-vehicle-taste mixtures. Subjects made estimates of the sweetness and bitterness of 27 stimuli. Sucrose (292, 585, and 1170 mM), caffeine (13, 26, and 52 mM), and binary mixtures of low (292-13 mM), middle (585-26 mM), and high (1170-52 mM) levels of both components were dispersed in water, carboxymethylcellulose (CMC) 1% w/v, and gelatin 6% w/v. The sweetness and bitterness of the sucrose-vehicle-caffeine combinations were significantly weaker than the respective sucrose-vehicle and caffeine-vehicle combinations. The emerged mutual suppressive effects were asymmetrical and persisted when both tastants were presented in CMC and gelatin. Moreover, the increase in vehicle consistency and the simultaneous addition of another taste reduced the perceived intensity of a taste either presented alone or dissolved in water. For both sweetness and bitterness, the total taste suppression observed was always significant.

Glycation of blood proteins during pregnancy and lactation in the rat.

Non-enzymatic glycation of blood proteins is a time and concentration dependent process and has been used clinically to monitor carbohydrate metabolism during human pregnancy. Since gestation in rats is of much shorter duration than in humans (3 weeks vs 9 mos) the question was raised whether similar differences in glycated proteins could be observed. Therefore, levels of glucose, glycated hemoglobin and fructosamine were measured during normal pregnancy and lactation in rats. Glucose levels during late pregnancy were significantly lower than in non pregnant and early pregnant rats. During lactation glucose levels return to normal. Glycated hemoglobin paralleled glucose decrease during late pregnancy and increased during lactation. Fructosamine followed a similar pattern. Therefore glycated hemoglobin and fructosamine appear to be reliable indicators of glucose status during gestation and lactation similar to humans and may have value as predictors of gestational diabetes mellitus once a suitable rat model is developed.

Differential growth requirements of several Leishmania spp. in chemically defined culture media.

17 strains of Leishmania from 4 species: brasiliensis, mexicana, donovani and garnhami have been continually cultured at 26 degrees C, in the absence of proteins, in a medium containing salts, glucose, D-ribose, 2-deoxyribose, hemin, tricine, HEPES, 34 amino acids and intermediates of amino acid metabolism, 23 vitamins, 6 nucleotides and tetrahydrofolic acid. A wide variation in growth requirements was observed among leishmaniae which permitted the preparation of different minimum culture media for each Leishmania spp. Virulence of parasites was maintained after 30 passages in these chemically defined media. The requirements for differentiation to amastigotes also varied among the species as a function of the temperature of incubation and the protein content of the culture medium. Bovine serum albumin tryptic peptides substituted fetal bovine serum as growth factors at 30-34 degrees C.

Protein and nucleotide contamination of bovine liver catalase used in culture medium explains growth of Trypanosoma cruzi.

Commercially available bovine liver catalase has been used to supplement chemically defined medium for growth of Trypanosoma cruzi. The protein extract was found to be contaminated with 25 to 30 protein bands as well as DNA and RNA polymers.