Presence of vascular endothelial growth factor during the first half of IVM improves the meiotic and developmental competence of porcine oocytes from small follicles

The aim of the present study was to investigate the effect of vascular endothelial growth factor (VEGF) on the meiotic and developmental competence of porcine oocytes from small follicles (SF; 0.5-3mm diameter). When cumulus-oocyte complexes (COCs) from medium-sized follicles (MF; 3-6mm diameter) and SF were cultured for IVM, the maturation rates were significantly higher for oocytes from MF than SF. Concentrations of VEGF in the medium were significantly higher for COCs cultured from MF than SF. When COCs from SF were exposed to 200ngmL-1 VEGF during the first 20h of IVM, the maturation rate improved significantly and was similar to that of oocytes derived from MF. The fertilisability of oocytes was also significantly higher than that of VEGF-free SF controls. Following parthenogenetic activation, the blastocyst formation rate improved significantly when SF COC culture was supplemented with 200ngmL-1 VEGF, with the rate similar to that of oocytes from MF. The results of the present study indicate that VEGF markedly improves the meiotic and developmental competence of oocytes derived from SF, especially at a concentration of 200ngmL-1 during the first 20h of IVM

Differential susceptibility in youth: evidence that 5-HTTLPR x positive parenting is associated with positive affect ‘for better and worse'

Positive affect has been implicated in the phenomenological experience of various psychiatric disorders, vulnerability to develop psychopathology and overall socio-emotional functioning. However, developmental influences that may contribute to positive affect have been understudied. Here, we studied youths' 5-HTTLPR genotype and rearing environment (degree of positive and supportive parenting) to investigate the differential susceptibility hypothesis (DSH) that youth carrying short alleles of 5-HTTLPR would be more influenced and responsive to supportive and unsupportive parenting, and would exhibit higher and lower positive affect, respectively. Three independent studies tested this gene–environment interaction (GxE) in children and adolescents (age range 9–15 years; total N=1874). In study 1 (N=307; 54% girls), positive/supportive parenting was assessed via parent report, in study 2 (N=197; 58% girls) via coded observations of parent–child interactions in the laboratory and in study 3 (N=1370; 53% girls) via self report. Results from all the three studies showed that youth homozygous for the functional short allele of 5-HTTLPR were more responsive to parenting as environmental context in a ‘for better and worse' manner. Specifically, the genetically susceptible youth (that is, S'S' group) who experienced unsupportive, non-positive parenting exhibited low levels of positive affect, whereas higher levels of positive affect were reported by genetically susceptible youth under supportive and positive parenting conditions. These GxE findings are consistent with the DSH and may inform etiological models and interventions in developmental psychopathology focused on positive emotion, parenting and genetic susceptibility

Anti-seizure therapy with a long-term, implanted intra-cerebroventricular delivery system for drug-resistant epilepsy: A first-in-man study

Background: A clinical feasibility study was undertaken at a single center of long-term intra-cerebroventricular drug delivery of the anti-seizure medication valproic acid, into the cerebrospinal fluid (CSF) in order to treat drug resistant focal seizures, using an implantable infusion system. The primary objective was to establish the dose range of VPA administered in this manner. Secondarily, safety, pharmacokinetics (PK) and a preliminary estimate of effectiveness were evaluated. Methods: In this single arm study, five adult subjects, with 29-234 focal onset seizures per month from a seizure focus involving the mesial temporal lobe were implanted with the system (clinicaltrials.gov identifier NCT02899611). Oral valproic acid (VPA) had previously been ineffective in all subjects. Post-surgery, pharmacokinetic studies of CSF infused VPA were performed. Valproic acid doses were increased stepwise in a standardised protocol. Findings: The procedure and implantation were well-tolerated by all subjects. Four subjects responded with > 50% seizure reduction at the highest tested dose of 160 mg/day. Two subjects experienced extended periods of complete seizure freedom. All five subjects reported significant quality of life improvement. No clinical dose limiting side effects were encountered and there was no evidence of local periventricular toxicity in three subjects who were evaluated with imaging (T2 MRI). Side effects included nausea and appetite loss but were not dose-limiting. Mean CSF valproic acid levels were 45 μg per ml (range 20-120 μg per ml), with corresponding serum levels of 4-14 μg per ml.  Subjects have received therapy for up to 2.5 years in total . The efficacy analysis presented focuses on the period of time with the current pump with a mean 12.5 months, range 11.5-15 months. Pump failure requiring reimplantation was a significant initial issue in all subjects but resolved with use of pumps suitably compatible with long-term exposure to valproic acid. Interpretation: The study demonstrated that chronic intraventricular administration of valproic acid is safe and effective in subjects with medically refractory epilepsy over many months. The procedure for implanting the infusion system is safe and well-tolerated. High CSF levels are achieved with corresponding low serum levels and this therapy is shown to be effective despite unsuccessful earlier use of oral valproate preparations. Drug side effects were minimal. Funding: The study was funded by Cerebral Therapeutics Inc., Suite 137 12635 East Montview Blvd Aurora CO 80045

Medulloblastoma Exosome Proteomics Yield Functional Roles for Extracellular Vesicles

<div><p>Medulloblastomas are the most prevalent malignant pediatric brain tumors. Survival for these patients has remained largely the same for approximately 20 years, and our therapies for these cancers cause significant health, cognitive, behavioral and developmental sequelae for those who survive the tumor and their treatments. We obviously need a better understanding of the biology of these tumors, particularly with regard to their migratory/invasive behaviors, their proliferative propensity, and their abilities to deflect immune responses. Exosomes, virus-sized membrane vesicles released extracellularly from cells after formation in, and transit thru, the endosomal pathway, may play roles in medulloblastoma pathogenesis but are as yet unstudied in this disease. Here we characterized exosomes from a medulloblastoma cell line with biochemical and proteomic analyses, and included characterization of patient serum exosomes. Further scrutiny of the proteomic data suggested functional properties of the exosomes that are relevant to medulloblastoma tumor biology, including their roles as proliferation stimulants, their activities as attractants for tumor cell migration, and their immune modulatory impacts on lymphocytes. Aspects of this held true for exosomes from other medulloblastoma cell lines as well. Additionally, pathway analyses suggested a possible role for the transcription factor hepatocyte nuclear factor 4 alpha (HNF4A); however, inhibition of the protein’s activity actually increased D283MED cell proliferation/clonogenecity, suggesting that HNF4A may act as a tumor suppressor in this cell line. Our work demonstrates that relevant functional properties of exosomes may be derived from appropriate proteomic analyses, which translate into mechanisms of tumor pathophysiology harbored in these extracellular vesicles.</p> </div

D283MED exosomes affect interferon-gamma output from activated PBMCs.

<p>IPA Networks 3 and 8 in combination (<b>A</b>) suggested that exosomes may be involved in immune cell cytokine release (boxed terms and scores for each work are show at right). Networks are represented as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042064#pone-0042064-g005" target="_blank"><b>Figures 5</b></a><b>, </b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042064#pone-0042064-g006" target="_blank"><b>6</b></a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042064#pone-0042064-g007" target="_blank"><b>7</b></a>. Immune-related cytokines are also noted in larger font and with gray fill. (<b>B</b>) shows exosome-induced changes in PHA-activated PBMCs; healthy donor PBMCs were stimulated with PHA (5 µg/ml) for 48 hrs and D283MED exosomes at the concentrations listed were added as well. Interferon-γ release was measured by ELISA. Differences between groups were statistically evaluated by ANOVA; significant differences (p<0.05) between groups are indicated by different “star cluster” numbers (eg, *, **, ***).</p

Categorization of D283MED exosomal proteins by subcellular localization and by function.

<p>The proteins listed in Table S1 were categorized as percentages of the total number of proteins identified using Ingenuity Pathway Analysis descriptions and literature searches. Proteins are classified by subcellular (or extracellular) localization (<b>A</b>) or by function (<b>B</b>).</p

Inhibition of hepatocyte nuclear factor 4α (HNF4A) actually increases D283MED cell growth.

<p>IPA Networks 3 and 5 in combination (<b>A</b>) reveal that HNF4A (circled in red) sits at a node of interaction with nearly a dozen other proteins in networks tied to cancer cell metabolism (boxed terms and scores for each work are show at right). Networks are represented as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042064#pone-0042064-g005" target="_blank"><b>Figures 5</b></a><b>, </b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042064#pone-0042064-g006" target="_blank"><b>6</b></a><b>, </b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042064#pone-0042064-g007" target="_blank"><b>7</b></a><b>and </b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042064#pone-0042064-g008" target="_blank"><b>8</b></a>. (<b>B</b>) D283MED cells were treated with MEDICA16 (125 µM) to inhibit HNF4A; cells were also treated with D283MED exosomes (25 or 100 µg/ml) with or without MEDICA16. Cells were grown in a clonogenic assay for 8 days and were quantified following the various treatments. Differences between groups were statistically evaluated by ANOVA; significant differences (p<0.05) between groups are indicated by different “star cluster” numbers (eg, *, **, ***). Control cell growth (D283MED cells with no drug or exosome treatments) was defined as 100%.</p