A protocol to assess cell cycle and apoptosis in human and mouse pluripotent cells

Helicobacter pylori is a highly successful pathogen uniquely adapted to colonize humans. Gastric infections with this bacterium can induce pathology ranging from chronic gastritis and peptic ulcers to gastric cancer. More virulent H. pylori isolates harbour numerous well-known adhesins (BabA/B, SabA, AlpA/B, OipA and HopZ) and the cag (cytotoxin-associated genes) pathogenicity island encoding a type IV secretion system (T4SS). The adhesins establish tight bacterial contact with host target cells and the T4SS represents a needle-like pilus device for the delivery of effector proteins into host target cells such as CagA. BabA and SabA bind to blood group antigen and sialylated proteins respectively, and a series of T4SS components including CagI, CagL, CagY and CagA have been shown to target the integrin β₁ receptor followed by injection of CagA across the host cell membrane. The interaction of CagA with membrane-anchored phosphatidylserine may also play a role in the delivery process. While substantial progress has been made in our current understanding of many of the above factors, the host cell receptors for OipA, HopZ and AlpA/B during infection are still unknown. Here we review the recent progress in characterizing the interactions of the various adhesins and structural T4SS proteins with host cell factors. The contribution of these interactions to H. pylori colonization and pathogenesis is discussed.     

Sensorimotor Plasticity after Music-Supported Therapy in Chronic Stroke Patients Revealed by Transcranial Magnetic Stimulation

Background

Several recently developed therapies targeting motor disabilities in stroke sufferers have shown to be more effective than standard neurorehabilitation approaches. In this context, several basic studies demonstrated that music training produces rapid neuroplastic changes in motor-related brain areas. Music-supported therapy has been recently developed as a new motor rehabilitation intervention.

Methods and Results

In order to explore the plasticity effects of music-supported therapy, this therapeutic intervention was applied to twenty chronic stroke patients. Before and after the music-supported therapy, transcranial magnetic stimulation was applied for the assessment of excitability changes in the motor cortex and a 3D movement analyzer was used for the assessment of motor performance parameters such as velocity, acceleration and smoothness in a set of diadochokinetic movement tasks. Our results suggest that the music-supported therapy produces changes in cortical plasticity leading the improvement of the subjects'' motor performance.

Conclusion

Our findings represent the first evidence of the neurophysiological changes induced by this therapy in chronic stroke patients, and their link with the amelioration of motor performance. Further studies are needed to confirm our observations.     

Increase in antioxidant and antihypertensive activity by Oenococcus oeni in a yeast autolysis wine model

Accelerated autolysis of Saccharomyces cerevisiae mc2 in synthetic wine medium enabled the release of 3.7 mg peptide nitrogen/l, concomitantly with an increase in antioxidant properties (243 μmol FeSO₄/l in the case of ferric reducing antioxidant power and 0.5% in 2,2-diphenyl-1-picrylhydrazyl radical scavenging) and antihypertensive activity (22% in angiotensin I-converting enzyme inhibitory activity). Sequential inoculation of a proteolytic Oenococcus oeni strain in the synthetic medium after yeast autolysis produced an increase in peptide nitrogen concentration of 1.5 mg/l after 48 h of growth. After this incubation time an improvement in antihypertensive and antioxidant activities was detected. Oenococcus oeni X₂L could give additional value to wine because of the bioactive peptides with multifunctional beneficial activity released as consequence of its proteolytic activity.     

A novel approach using transcomplementing adenoviral vectors for gene therapy of adrenocortical cancer.

Current therapies for adrenocortical carcinomas do not improve the life expectancy of patients. In this study, we tested whether a gene-transfer therapy based upon a suicide gene/prodrug system would be effective in an animal model of the disease. We employed E4- and E1A/B-depleted, herpes simplex virus-thymidine kinase-expressing adenoviral mutants that transcomplement each other within tumor cells, hereby improving transgene delivery and efficacy by viral replication in situ. Transcomplementation of vectors increased the fraction of transduced of tumor cells. This increase was accompanied by greater tumor volume reduction compared to non-transcomplementing approaches. Survival time improved with non-replicating vectors plus GCV compared to controls. However, transcomplementation/replication of vectors led to a further significant increment in anti-tumor activity and survival time (p < 0.02). In treated animals, we observed a high number of apoptotic nuclei both adjacent to and distant from injection sites and sites of viral oncolysis. Ultrastructural analyses exhibited nuclear inclusion bodies characteristic of virus production in situ, and provided further evidence that this therapy induced apoptotic cell death within tumor cells. We conclude that the efficacy of suicide gene therapy is significantly amplified by viral replication and, in combination with GCV, significantly reduces tumor burden and increases survival time.     

The brain glutamate system in liver failure

Liver failure results in significant alterations of the brain glutamate system. Ammonia and the astrocyte play major roles in such alterations, which affect several components of the brain glutamate system, namely its synthesis, intercellular transport (uptake and release), and function. In addition to the neurological symptoms of hepatic encephalopathy, modified glutamatergic regulation may contribute to other cerebral complications of liver failure, such as brain edema, intracranial hypertension and changes in cerebral blood flow. A better understanding of the cause and precise nature of the alterations of the brain glutamate system in liver failure could lead to new therapeutic avenues for the cerebral complications of liver disease.     

A selective differential medium for Lactobacillus plantarum

The quantification of exogenous lactobacilli in faecal samples is frequently required for the evaluation of the intestinal colonization by probiotic bacteria. In this study, a selective and differential medium, designated LPSM, was developed for the culture of exogenous Lactobacillus plantarum. In quantitative assays, LPSM showed a sensitivity similar to those of enriched and Lactobacillus-adapted media. The presence of ciprofloxacin made LPSM inhibitory to most intestinal bacteria, including endogenous acid lactic bacteria, whereas exogenous L. plantarum strains grew producing a yellow color caused by acid production from sorbitol in the presence of bromocresol purple. The results showed that LPSM is suitable for detection and enumeration of L. plantarum in faecal samples.     

Variations in airborne pollen in central and south-western Spain in relation to the distribution of potential sources

The present study seeks to compare daily and hourly airborne pollen concentrations at eight different stations in Castilla-La Mancha (central Spain) and Extremadura (south-western Spain) and assess pollen distribution sources. Sampling stations were located 69–440 km apart in a straight line in Albacete, Toledo, Talavera de la Reina and Ciudad Real in Castilla-La Mancha, and in Badajoz, Plasencia, Santa Amalia and Zafra in Extremadura. Airborne pollen was collected using Hirst-type volumetric spore traps. Quercus was the most abundant pollen type at all stations except for Ciudad Real, where Olea pollen predominated. Comparisons of daily data between pairs of stations revealed statistically significant positive correlations in all cases for Poaceae and Olea. Comparisons of hourly data between stations indicated greater differences than daily data. Analysis of correlation coefficients and straight-line distances between stations revealed a strong negative correlation. Analysis of total airborne pollen data for the eight sampling stations suggests that airborne pollen concentrations decrease from west to east and from south to north, partly reflecting dominant wind patterns. A clear correlation was observed between airborne pollen concentrations and the surface area covered by olive crops in a 50 km radius around the sampling stations.