Telomerase expression in B-cell chronic lymphocytic leukemia predicts survival and delineates subgroups of patients with the same igVH mutation status and different outcome.

Activation of telomerase reverse transcriptase (hTERT) is essential for unlimited cell growth and plays a critical role in tumorigenesis. We investigated hTERT gene expression in 134 B-cell chronic lymphocytic leukemia (B-CLL) cases and evaluated its prognostic value with other prognostic markers (IgVH mutation status, CD38 and ZAP-70 expression). Real-time PCR assays to quantify either all hTERT transcripts (AT) or only the full length (FL) transcript encoding the functional protein were developed. hTERT-AT levels strongly correlated with hTERT-FT levels (r=0.743, P<0.0001); both inversely correlated with the percentage of IgVH mutation (P<0.005) and were significantly higher in unmutated than in mutated cases (P=0.004 and P=0.001, respectively). The hTERT values which best discriminated between the unmutated and mutated IgVH cases were 150 and 40 copies for hTERT-AT and hTERT-FL, respectively. Using these cut-off values, there was a significant difference in the survival of patients with high or low hTERT levels (P<0.0001). Unmutated cases with low hTERT levels had an overall survival close to mutated cases with high hTERT levels. Thus, this work identifies hTERT-RNA level as a new prognostic marker in B-CLL, and may be used to identify previously unrecognized patient groups with the same IgVH mutation status and different disease outcomes.

Association between SNAP-25 gene polymorphisms and cognition in autism: functional consequences and potential therapeutic strategies.

Synaptosomal-associated protein of 25 kDa (SNAP-25) is involved in different neuropsychiatric disorders, including schizophrenia and attention-deficit/hyperactivity disorder. Consistently, SNAP-25 polymorphisms in humans are associated with hyperactivity and/or with low cognitive scores. We analysed five SNAP-25 gene polymorphisms (rs363050, rs363039, rs363043, rs3746544 and rs1051312) in 46 autistic children trying to correlate them with Childhood Autism Rating Scale and electroencephalogram (EEG) abnormalities. The functional effects of rs363050 single-nucleotide polymorphism (SNP) on the gene transcriptional activity, by means of the luciferase reporter gene, were evaluated. To investigate the functional consequences that SNAP-25 reduction may have in children, the behaviour and EEG of SNAP-25(+/-) adolescent mice (SNAP-25(+/+)) were studied. Significant association of SNAP-25 polymorphism with decreasing cognitive scores was observed. Analysis of transcriptional activity revealed that SNP rs363050 encompasses a regulatory element, leading to protein expression decrease. Reduction of SNAP-25 levels in adolescent mice was associated with hyperactivity, cognitive and social impairment and an abnormal EEG, characterized by the occurrence of frequent spikes. Both EEG abnormalities and behavioural deficits were rescued by repeated exposure for 21 days to sodium salt valproate (VLP). A partial recovery of SNAP-25 expression content in SNAP-25(+/-) hippocampi was also observed by means of western blotting. A reduced expression of SNAP-25 is responsible for the cognitive deficits in children affected by autism spectrum disorders, as presumably occurring in the presence of rs363050(G) allele, and for behavioural and EEG alterations in adolescent mice. VLP treatment could result in novel therapeutic strategies.

Reduced SNAP-25 increases PSD-95 mobility and impairs spine morphogenesis.

Impairment of synaptic function can lead to neuropsychiatric disorders collectively referred to as synaptopathies. The SNARE protein SNAP-25 is implicated in several brain pathologies and, indeed, brain areas of psychiatric patients often display reduced SNAP-25 expression. It has been recently found that acute downregulation of SNAP-25 in brain slices impairs long-term potentiation; however, the processes through which this occurs are still poorly defined. We show that in vivo acute downregulation of SNAP-25 in CA1 hippocampal region affects spine number. Consistently, hippocampal neurons from SNAP-25 heterozygous mice show reduced densities of dendritic spines and defective PSD-95 dynamics. Finally, we show that, in brain, SNAP-25 is part of a molecular complex including PSD-95 and p140Cap, with p140Cap being capable to bind to both SNAP-25 and PSD-95. These data demonstrate an unexpected role of SNAP-25 in controlling PSD-95 clustering and open the possibility that genetic reductions of the protein levels - as occurring in schizophrenia - may contribute to the pathology through an effect on postsynaptic function and plasticity.

Energy expenditure of critically ill neonatal foals.

REASONS FOR PERFORMING STUDY: Nutritional support in critically ill neonatal foals is of great importance given their high metabolic rate and minimal stores of energy and protein. Nutrient requirements of healthy growing foals have been estimated based on daily milk intake; however, little is known about the resting energy expenditure (REE) of sick foals. OBJECTIVES: To determine REE in critically ill neonatal foals (sepsis and/or hypoxic-ischaemic encephalopathy [HIE] and compare this with REE in control foals. METHODS: Critically ill newborn foals admitted to the Fundació Hospital Clinic Veterinari, Universitat Autònoma de Barcelona, Spain from March 2009 to February 2011 were included in this study. Healthy neonatal foals and foals with nonsystemic conditions were used as controls. Oxygen consumption and CO2 production were measured with a respiratory monitor connected to a tight fitting facemask and REE (kcal/kg bwt/day) was calculated with the abbreviated Weir formula. Measurements were performed within 24 h of admission and repeatedly during hospitalisation. RESULTS: Twenty-seven foals were included (16 critically ill foals and 11 controls) and a total of 47 measurements were performed. In the critically ill, REE was reduced (mean +/- s.e. 49.5 +/- 2.1 kcal/kg bwt/day) on admission relative to the controls. In surviving foals (n = 5), REE before hospital discharge was not different (68.4 +/- 7.0 kcal/kg bwt/day) from control foals (64.8 +/- 2.7 kcal/kg bwt/day). CONCLUSIONS: REE was lower in critically ill foals upon admission (40-50 kcal/kg bwt/day) and normalised before hospital discharge (60-80 kcal/kg bwt/day). POTENTIAL RELEVANCE: Critically ill neonatal foals tolerating enteral feeding would receive approximately their REE when given 10% of their bodyweight in mare's milk daily. For sick neonates unable to tolerate enteral nutrition, provision of 50 kcal/kg bwt/day would be a reasonable goal for parenteral nutrition.