Histone H1 dephosphorylation is not a general feature in early apoptosis

Histone H1 is a family of nucleosomal proteins that exist in a number of subtypes. These subtypes can be modified after translation in various ways, above all by phosphorylation. Increasing levels of H1 phosphorylation has been correlated with cell cycle progression, while both phosphorylation and dephosphorylation of histone H1 have been linked to the apoptotic process. Such conflicting results may depend on which various apoptosis-inducing agents cause apoptosis via different apoptotic pathways and often interfere with cell proliferation. Therefore, we investigated the relation between apoptosis and H1 phosphorylation in Jurkat cells after apoptosis induction via both the extrinsic and intrinsic pathways and by taking cell cycle effects into account. After apoptosis induction by anti-Fas, no significant dephosphorylation, as measured by capillary electrophoresis, or cell cycle-specific effects were detected. In contrast, H1 subtypes were rapidly dephosphorylated when apoptosis was induced by camptothecin. We conclude that histone H1 dephosphorylation is not connected to apoptosis in general but may be coupled to apoptosis by the intrinsic pathway or to concomitant growth inhibitory signaling.     

Environmental enrichment,sexual dimorphism,and brain size in sticklebacks

Evidence for phenotypic plasticity in brain size and the size of different brain parts is widespread, but experimental investigations into this effect remain scarce and are usually conducted using individuals from a single population. As the costs and benefits of plasticity may differ among populations, the extent of brain plasticity may also differ from one population to another. In a common garden experiment conducted with three‐spined sticklebacks (Gasterosteus aculeatus) originating from four different populations, we investigated whether environmental enrichment (aquaria provided with structural complexity) caused an increase in the brain size or size of different brain parts compared to controls (bare aquaria). We found no evidence for a positive effect of environmental enrichment on brain size or size of different brain parts in either of the sexes in any of the populations. However, in all populations, males had larger brains than females, and the degree of sexual size dimorphism (SSD) in relative brain size ranged from 5.1 to 11.6% across the populations. Evidence was also found for genetically based differences in relative brain size among populations, as well as for plasticity in the size of different brain parts, as evidenced by consistent size differences among replicate blocks that differed in their temperature.     

Expression of an HCV core antigen coding gene in tobacco (N. tabacum L.)

Hepatitis C virus (HCV) is the major agent causing chronic liver disease. The core gene is the most conserved sequence in the HCV genome and proved immunoreactive when expressed in bacteria and antigenic in humans. In order to test the ability of plants to express the core gene for the production of core antigen, transgenic tobacco plants carrying the core gene were generated. The core protein was stably synthesized in T(0) and T(1) generations and was found to be immunoreactive, not only with anti-core polyclonal and monoclonal antibodies, but also was able to recognize the HCV virus in infected human serum. The prospects of producing a plant based vaccine and/or a food vaccine for this important virus are discussed.     

Toxoplasmosis in pregnant women in Crete

In Greece there are no official recommendations concerning the management of pregnant women for the prevention of congenital toxoplasmosis. A protocol for monitoring pregnant women was designed in order to differentiate between acute and latent toxoplasmosis and was tested successfully for 7 years. The maternofetal transmission rate in Crete was assessed and a map showing seroprevalence of pregnant women in all prefectures of Greece was prepared. The high seroprevalence of Toxoplasma gondii in Greece (up to 46% in some areas) may be explained by: (a) the presence of a great number of stray cats; (b) the Greek diet consisting of large amounts of raw, wild vegetables and salads that could easily be contaminated with oocysts; (c) the high consumption of meat, smoked pork and sausages, well-documented sources of T. gondii infection. T. gondii genotypes were characterized, directly from clinical samples, after PCR-RFLP on the SAG2 gene and sequence analysis at the restriction sites. They belonged to all 3 clonal lineages.     

Absorption of intact proteins by the intestinal epithelium of trout,Salmo gairdneri

In the rainbow trout, a carnivorous species with a stomach, orally administered horseradish peroxidase (HRP) crossed the intestinal epithelium into the circulation. The protein first appeared in the blood 7 to 8 h after intubation, and could be assayed up to 24 h after administration. The method used, which combines ELISA (enzyme-linked immunoabsorbant assay) and chemiluminescence, enabled the transfer to be measured quantitatively. There was a direct correlation between the quantity ingested and the quantity transferred to the plasma within the experimental limits chosen. The clearance was monophasic and exponential (clearance rate: 3% per minute). Up to 6% of the ingested HRP was transferred to the blood. By cytochemistry it was possible to demonstrate that the protein crossed the intestinal cells at the level of the posterior segment, escaping the particularly intense intracellular lysosomal digestion. After entering the intercellular space, HRP was transferred to the interstitial space of the subepithelial lamina propria. During this transfer the HRP was in close contact with infiltrated macrophages and leukocytes resembling lymphoid cells. Thus, the passage of these protein particles could be the first indispensable step in the possible triggering of a local and/or a systemic immune response.     

Immunological responses in cancer patients after vaccination with the therapeutic telomerase-specific vaccine Vx-001

Vx-001, an HLA-A*0201 restricted telomerase (TERT)-specific anti-tumor vaccine, is composed of the 9-mer cryptic TERT₅₇₂ peptide and its optimized variant TERT_572Y. We have previously shown that Vx-001 is non-toxic, highly immunogenic and in vaccinated NSCLC patients early specific immune response is associated with prolonged survival. The aim of the present study was to investigate the specific T-cell immune response against Vx-001. Fifty-five patients with chemo-resistant advanced solid tumors were vaccinated with TERT_572Y (2 subcutaneous injections) followed by TERT₅₇₂ peptide (4 subcutaneous injections) every 3 weeks. Specific immune response was evaluated by IFN-γ and perforin ELISpot and intracellular cytokine staining assays. TERT-reactive T cells were detected in 27 (51%) out of 53 evaluable patients after the 2nd vaccination and in 22 (69%) out of 32 evaluable patients after the completion of 6 vaccinations. Immune responses developed irrespective of the stage of disease and disease status before vaccination. Patients with disease progression at study entry who developed a post-vaccination-induced immunological response had a significant overall survival benefit compared to the post-vaccination non-responders. The Vx-001 vaccine is a promising candidate for cancer immunotherapy since it can induce a TERT-specific T-cell immune response that is associated with prolonged survival.     

Mutations in ANO3 Cause Dominant Craniocervical Dystonia: Ion Channel Implicated in Pathogenesis

In this study, we combined linkage analysis with whole-exome sequencing of two individuals to identify candidate causal variants in a moderately-sized UK kindred exhibiting autosomal-dominant inheritance of craniocervical dystonia. Subsequent screening of these candidate causal variants in a large number of familial and sporadic cases of cervical dystonia led to the identification of a total of six putatively pathogenic mutations in ANO3, a gene encoding a predicted Ca²⁺-gated chloride channel that we show to be highly expressed in the striatum. Functional studies using Ca²⁺ imaging in case and control fibroblasts demonstrated clear abnormalities in endoplasmic-reticulum-dependent Ca²⁺ signaling. We conclude that mutations in ANO3 are a cause of autosomal-dominant craniocervical dystonia. The locus DYT23 has been reserved as a synonym for this gene. The implication of an ion channel in the pathogenesis of dystonia provides insights into an alternative mechanism that opens fresh avenues for further research.     

Fetal-specific DNA methylation ratio permits noninvasive prenatal diagnosis of trisomy 21

The trials performed worldwide toward noninvasive prenatal diagnosis (NIPD) of Down's syndrome (or trisomy 21) have shown the commercial and medical potential of NIPD compared to the currently used invasive prenatal diagnostic procedures. Extensive investigation of methylation differences between the mother and the fetus has led to the identification of differentially methylated regions (DMRs). In this study, we present a strategy using the methylated DNA immunoprecipitation (MeDiP) methodology in combination with real-time quantitative PCR (qPCR) to achieve fetal chromosome dosage assessment, which can be performed noninvasively through the analysis of fetal-specific DMRs. We achieved noninvasive prenatal detection of trisomy 21 by determining the methylation ratio of normal and trisomy 21 cases for each tested fetal-specific DMR present in maternal peripheral blood, followed by further statistical analysis. The application of this fetal-specific methylation ratio approach provided correct diagnosis of 14 trisomy 21 and 26 normal cases.