Molecular role of the PAX5-ETV6 oncoprotein in promoting B-cell acute lymphoblastic leukemia

PAX5 is a tumor suppressor in B-ALL, while the role of PAX5 fusion proteins in B-ALL development is largely unknown. Here, we studied the function of PAX5-ETV6 and PAX5-FOXP1 in mice expressing these proteins from the Pax5 locus. Both proteins arrested B-lymphopoiesis at the pro-B to pre-B-cell transition and, contrary to their proposed dominant-negative role, did not interfere with the expression of most regulated Pax5 target genes. Pax5-Etv6, but not Pax5-Foxp1, cooperated with loss of the Cdkna2a/b tumor suppressors in promoting B-ALL development. Regulated Pax5-Etv6 target genes identified in these B-ALLs encode proteins implicated in pre-B-cell receptor (BCR) signaling and migration/adhesion, which could contribute to the proliferation, survival, and tissue infiltration of leukemic B cells. Together with similar observations made in human PAX5-ETV6⁺ B-ALLs, these data identified PAX5-ETV6 as a potent oncoprotein that drives B-cell leukemia development.     

Structure and atomic fluctuation patterns of potato carboxypeptidase a inhibitor protein

Molecular dynamics (MD) simulation methods were applied to the study of the structural and dynamic fluctuation properties of the potato carboxypeptidase A inhibitor protein (PCI) immersed in a bath of 1259 water molecules. A trajectory of 200 ps was generated at constant temperature and pressure. The crystallographic structure of PCI, as found in its complex with bovine carboxy-peptidase A (CPA), was used to seed the MD simulation. Analyses show that the structure of the PCI core is fairly rigid and stable in itself, and that little deformation is caused by the protein-protein interactions found in the PCI-CPA complex. The N-terminal tail fluctuates to approach the core structure and appears as a relatively disordered region. In contrast, the conformations of the C-terminal tail, which is involved in the inhibitory mechanism, fluctuates in the neighborhood of the X-ray structure in orientations which facilitate CPA binding. Comparison with the structural entries for PCI in water obtained from both 2D-NMR experiments and X-ray data shows that important features of the MD structural results fluctuates between the initial crystal values and those obtained from the NMR solution structure. This fluctuation is not uniform; minor regions move away from the X-ray conformation while they do not approach the NMR conformation. The results reported suggest that the trajectory is long enough to show a behavior that is consistent with the conformational space available to the protein in solution.Abbreviations CPA Carboxypeptidase - DG Distance Geometry - NMR Nuclear Magnetic Resonance - NIS Non Inertial Solvent - MD Molecular Dynamics - PBC Periodic Boundary Conditions - PCI Potato Carboxypeptidase Inhibitor - RMSD Root Mean Square Deviation - a.m.u. Atomic mass units Correspondence to: O. Tapia     

A novel cysteine proteinase inhibitor from seeds of Enterolobium contortisiliquum and its effect on Callosobruchus maculatus larvae

This study focused on the characterization of a novel cysteine proteinase inhibitor from Enterolobium contortisiliquum seeds targeting the inhibition of the growth of Callosobruchus maculatus larvae, an important cosmopolitan pest of the cowpea Vigna unguiculata during storage. The inhibitor was isolated by ion-exchange besides of size exclusion chromatography. EcCI molecular mass is 19,757 Da, composed of two polypeptide chains. It strongly inhibits papain (Ki_app 0.036 nM) and proteinases from the midguts of C. maculatus (80 μg mL^?1, 60% inhibition). The inhibitory activity is reduced by 40% after a heat treatment at 100 °C for 2 h. The protein displayed noxious activity at 0.5% and 1% (w/w) when incorporated in artificial seeds, reducing larval mass in 87% and 92%, respectively. Treatment of C. maculatus larvae with conjugated EcCI-FIT and subsequent biodistribution resulted in high fluorescence intensity in midguts and markedly low intensity in malpighian tubules and fat body. Small amounts of labeled proteins were detected in larvae feces. The detection of high fluorescence in larvae midguts and low fluorescence in their feces indicate the retention of the FITC conjugated EcCI inhibitor in larvae midguts. These results demonstrate the potential of the natural protein from E. contortisiliquum to inhibit the development of C. maculatus.     

A new species of Proctoeces and reinstatement of Proctoeces humboldti George-Nascimento and Quiroga 1983 (Digenea: Fellodistomidae) based on molecular and morphological evidence

The most studied digenean of marine organisms in Chile is by far Proctoeces humboldti, a parasite of the intestine of the clingfish Sicyases sanguineus and gonad of the keyhole limpet Fissurella spp. (progenetic metacercariae). The mussel Perumytilus purpuratus has been suggested as the first intermediate host for this digenean. In a study examining the parasites of S. sanguineus from central Chile, we found specimens of Proctoeces showing significant morphological differences with P. humboldti. To assist in the resolution of the taxonomic identification of these specimens, as well sporocysts obtained from the mussel P. purpuratus from central and northern Chile, phylogenetic studies using DNA sequences from the SSU rRNA, as well the LSU rRNA and Cox 1 gene were performed. Results showed that the clingfish S. sanguineus is a host for two species of Proctoeces (P. humboldti and P. syciases n. sp.) along the northern and central Chilean coast, without geographic separation; the mussel P. purpuratus is the first intermediate host for P. syciases n. sp. but not for P. humboldti in central and northern Chile. Fissurellids (Archaeogastropoda) along the Chilean coast harbor only progenetic stages of P. humboldti, but there is no evidence of progenesis for P. syciases. The reinstatement of Proctoeces humboldti is strongly suggested.     

Acquisition of thermotolerance in sea urchin embryos correlates with the synthesis and age of the heat shock proteins

Preheating at 31 degrees C induces thermotolerance in Paracentrotus lividus embryos, which therefore become able to withstand 1-h treatment at the otherwise lethal temperature of 35 degrees C, and to develop normally. The acquisition of thermotolerance is positively correlated with the amount of heat shock proteins produced during the 31 degrees C treatment. Evidence is provided that the heat shock proteins, although present in the embryo for long periods after synthesis, lose their effect on thermotolerance within 3 h of the cessation of synthesis.     

White matter fractional anisotropy is related to processing speed in metabolic syndrome patients: a case-control study

Background  

Metabolic Syndrome (MetSd) is a cluster of vascular risk factors that may influence cerebrovascular pathology during aging. Recently, microstructural white matter (WM) changes detected by diffusion tensor imaging (DTI) and processing speed deficits have been reported in MetSd patients. We aimed to test the relationship between WM alteration and cognitive impairment in these patients.     

Selected plasma biochemistry parameters in gilthead seabream (Sparus aurata) juveniles

The aim of this study was to assess plasma biochemistry parameters with the potential of being used as indicators of the nutritional status for healthy gilthead seabream juveniles. Triplicate groups of 18 seabream (body weight of 58 g) were kept unfed for 24 h, 7 or 14 days. Nine fish per treatment were then sampled randomly for blood collection and the following parameters analyzed in the plasma using standard clinical methods: glucose; protein; triglycerides; cholesterol; calcium; magnesium; inorganic phosphorus; alkaline phosphatase (ALP); aspartate aminotransferase (AST); lactate dehydrogenase (LDH); gamma‐glutamyl transferase (GGT); creatine phosphokinase (CPK); and lipase. Biochemical parameters showed lower variability among individuals than did enzymatic parameters. Plasma glucose, protein, cholesterol, calcium and inorganic phosphorus levels were inversely related to the duration of starvation. On the contrary, plasma triglycerides decreased significantly during the first week of starvation and remained stable in the second week. Plasma ALP, AST and LDH decreased significantly after 1 week of starvation and then remained constant. In healthy seabream juveniles, plasma glucose, protein, cholesterol, calcium and inorganic phosphorus are responsive to starvation and may be useful indicators of the nutritional status of the animals. Indicative baseline reference values for gilthead seabream juveniles starved for 24 h and held at optimum temperature are: protein, 3.7–4.9 g dl^?1; cholesterol, 341–407 mg dl^?1; calcium, 13.1–8.0 mg dl^?1; and inorganic phosphorus, 10–14.2 mg dl^?1. Plasma triglycerides, along with plasma enzyme activities, may be useful as indicators of short term starvation. For these parameters baseline values after 1 week of starvation were: triglycerides: 138–230 mg dl^?1; ALP: 58–125 U L^?1; AST: 15–127 U L^?1; and LDH 61–677 U L^?1. Plasma glucose is only responsive to longer starvation periods, remaining relatively stable during the first week of starvation, and ranging from 59 to 196 mg dl^?1.     

Gene expression modifications in the liver caused by binge drinking and S-adenosylmethionine feeding. The role of epigenetic changes

Chronic ethanol ingestion, achieved by feeding ethanol at a constant rate using intragastric tube feeding, alters the expression of genes in the liver. This is done by epigenetic mechanisms, which depend on the blood alcohol levels at the time of killing. However, acute bolus feeding of ethanol changes gene expression without lasting epigenetic changes. This occurs with histone 3 methylation and acetylation modifications. The gene expression response to an acute bolus of ethanol might be modified by feeding S-adenosylmethionine (SAMe), a methyl donor. In the present study, rats were given a bolus of ethanol (6 g/kg body weight (bw), SAMe (1 g/kg bw), ethanol + SAMe, or isocaloric glucose. The group of rats (n = 3) were killed at 3 and 12 h post bolus, and gene microarray analysis was performed on their liver cells. SAMe reduced the 3 h blood ethanol levels and increased the ALT levels at 3 h. Venn diagrams showed that alcohol changed the expression of 646 genes at 3 h post bolus and 586 genes at 12 h. SAMe changed the expression of 1,012 genes when fed with ethanol 3 h post ethanol bolus and 554 genes at 12 h post ethanol bolus. SAMe alone changed the expression of 1,751 genes at 3 h and 1,398 at 12 h. There were more changes in gene expression at 3 h than at 12 h post ethanol when ethanol alone was compared to the dextrose control. The same was true when SAMe was compared to SAMe + ethanol. Ethanol up regulated gene expression in most functional pathways at 3 h. However, when SAMe was fed with ethanol at 3 h, most pathways were down regulated. At 12 h, however, when ethanol was fed, the pathways were half up regulated and half down regulated. The same was true when SAMe + ethanol was fed. The expression of epigenetically important genes, such as BHMT and Foxn3, was up regulated 3 h post alcohol bolus. At 3 h, SAMe down regulated the expression of genes, such as BHMT, Mat2a, Jun, Tnfrs9, Ahcy 1, Tgfbr1 and 2, and Pcaf. At 12 h, the insulin signaling pathways were half down regulated by ethanol, which was partly prevented by SAMe. The MAPK pathway was up regulated by ethanol, but SAMe did not prevent this. In conclusion, profound changes in gene expression evolved between 3 h and 12 post ethanol bolus. SAMe down regulated these changes in gene expression at 3 h, and less so at 12 h.     

Divergence of the IGS rDNA in Fusarium proliferatum and Fusarium globosum reveals two strain specific non-orthologous types

A phylogenic analysis of Fusarium proliferatum and closely related species was performed using the most variable part within the intergenic spacer of the nuclear ribosomal DNA (IGS) and compared with a previously reported phylogeny performed in the same group of samples with a partial region of the nuclear single copy gene encoding the elongation factor 1α (EF-1α). The phylogenies from both genomic sequences were not concordant and revealed the presence of two non-orthologous IGS types, named types I and II, in F. proliferatum and Fusarium globosum.