Growth, Development and Survival of Holothuria scabra Larvae in Different Microalgal Regimens and Water Rearing Media

Different aspects of Holothuria scabra larval production, including feeding regimen and water treatment, were investigated under experimental conditions. This study highlights the optimization of techniques and simplification of the requirements of sea cucumber larval rearing. The growth performance, development, and survivorship of H. scabra larvae were measured to assess which treatment provides optimum results. Chaetoceros gracilis (Cgr) and Chaetoceros calcitrans (Cc) were administered singly and in combination (Cgr-Cc) to sea cucumber larvae. Growth was highest in combined Cgr-Cc feed with mean final length of 2088µm, followed by Cc with 1855 µm and Cgr with 1800 µm, but with no significant difference (p > 0.05). Similarly, survival rates among treatments were not statistically different (Cgr-Cc = 2.23%; Cgr = 1.6%; Cc = 1.3%) (p > 0.05). However, larval development was better in combined Cgr-Cc and Cc single diet, with 90% and 100% composition of early juveniles on Day 30. Slower development was observed in Cgr single feed, with only 90% early juveniles observed later on Day 35. Different microalgal concentration of Cgr-Cc (10,000, 30,000 and 50,000 cells.mL-1) were also tested. Juveniles (~3 mm) yielded from 50,000 cells.mL-1 microalgal concentrations were five times larger than when fed at 10,000 cells.mL-1 microalgae. Development of larvae was also faster in 50,000 cells.mL-1, yielding harvestable juveniles in 25 days. However, water replenishment in tanks with high microalgal density should also be regularly done at 50-70% rate in two days interval to mitigate fouling. In addition, sand-filtered, chlorinated, and UV-treated seawater were also tested for their efficiency as culture media. Growth rates were significantly highest in sand-filtered seawater (68.3 µm.d-1), followed by UV-treated seawater (52.4 µm.d-1), and by chlorinated seawater (34.8 µm.d-1) (p < 0.05). Larval development did not differ in sand-filtered and UV-treated seawater, yielding ~1 mm juveniles as early as Day 25. Likewise, sand-filtered seawater rendered highest survival of larvae (10.24%) followed by UV-treated seawater (6.24%); chlorinated seawater yielded lowest (2.60%) (p < 0.05). Although a sterilization process is advised, findings on sand-filtered seawater as a rearing medium were notable

Space-time correlations in inflationary spectra, a wave packet analysis

The inflationary mechanism of mode amplification predicts that the state of each mode with a given wave vector is correlated to that of its partner mode with the opposite vector. This implies nonlocal correlations which leave their imprint on temperature anisotropies in the cosmic microwave background. Their spatial properties are best revealed by using local wave packets. This analysis shows that all density fluctuations giving rise the large scale structures originate in pairs which are born near the reheating. In fact each local density fluctuation is paired with an oppositely moving partner with opposite amplitude. To obtain these results we first apply a ``wave packet transformation'' with respect to one argument of the two point correlation function. A finer understanding of the correlations is then reached by making use of coherent states. The knowledge of the velocity field is required to extract the contribution of a single pair of wave packets. Otherwise, there is a two-folded degeneracy which gives three aligned wave packets arising from two pairs. The applicability of these methods to observational data is briefly discussed.Comment: revised version, accepted for publication in PRD ; the complementarity and the usefullness of wave packet analysis have been emphasized ; 32 pages, 6 figure

Composition of Ragusano Cheese During Aging

Ragusano cheese is a brine-salted pasta filata cheese. Composition changes during 12 mo of aging were determined. Historically, Ragusano cheese has been aged in caves at 14 to 16 degrees C with about 80 to 90% relative humidity. Cheeses (n = 132) included in our study of block-to-block variation were produced by 20 farmhouse cheese makers in the Hyblean plain region of the Province of Ragusa in Sicily. Mean initial cheese block weight was about 14 kg. The freshly formed blocks of cheese before brine salting contained about 45.35% moisture, 25.3% protein, and 25.4% fat, with a pH of 5.25. As result of the brining and aging process, a natural rind forms. After 12 mo of aging, the cheese contained about 33.6% moisture, 29.2% protein, 30.0% fat, and 4.4% salt with a pH of 5.54, but block-to-block variation was large. Both soluble nitrogen content and free fatty acid (FFA) content increased with age. The pH 4.6 acetate buffer and 12% TCA-soluble nitrogen as a percentage of total nitrogen were 16 and 10.7%, respectively, whereas the FFA content was about 643 mg/100 g of cheese at 180 d. Five blocks of cheese were selected at 180 d for a study of variation within block. Composition variation within block was large; the center had higher moisture and lower salt in moisture content than did the outside. Composition variation within blocks favored more proteolysis and softer texture in the center

Refractive change following pseudophakic vitrectomy: a retrospective review

Background To assess the occurrence and magnitude of refractive change in pseudophakic eyes undergoing 20 gauge pars plana vitrectomy without scleral buckling and to investigate possible aetiological factors. Methods Retrospective case note review of 87 pseudophakic eyes undergoing 20 gauge pars plana vitrectomy for a variety of vitreo-retinal conditions over a three-year period. Anterior chamber depth (ACD) was measured before and after vitrectomy surgery in 32 eyes. Forty-three pseudophakic fellow eyes were used as controls. Results Eighty-seven eyes (84 patients) were included in the study. Mean spherical equivalent refraction prior to vitrectomy was -0.20 dioptres, which changed to a mean of -0.65 dioptres postoperatively (standard deviation of refractive change 0.59, range-2.13 to 0.75 dioptres) (p < 0.001). Sixty-one of the 87(70%) eyes experienced a myopic shift and 45(52%) eyes had a myopic shift of -0.5 dioptres or more. Mean fellow eye refraction was -0.19 dioptres preoperatively and -0.17 dioptres postoperatively (p = 0.14)(n = 37) Mean ACD preoperatively was 3.29 mm and postoperatively 3.27 mm (p = 0.53) (n = 32) and there was no significant change in ACD with tamponade use. Regression analysis revealed no statistically significant association between changes in anterior chamber depth, as well as a wide variety of other pre-, intra and postoperative factors examined, and the refractive change observed. Conclusion Significant refractive changes occur in some pseudophakic patients undergoing 20 g pars plana vitrectomy. The mean change observed was a small myopic shift but the range was large. The aetiology of the refractive change is uncertain

Next-generation sequencing reveals large connected networks of intra-host HCV variants

Background: Next-generation sequencing (NGS) allows for sampling numerous viral variants from infected patients. This provides a novel opportunity to represent and study the mutational landscape of Hepatitis C Virus (HCV) within a single host. Results: Intra-host variants of the HCV E1/E2 region were extensively sampled from 58 chronically infected patients. After NGS error correction, the average number of reads and variants obtained from each sample were 3202 and 464, respectively. The distance between each pair of variants was calculated and networks were created for each patient, where each node is a variant and two nodes are connected by a link if the nucleotide distance between them is 1. The work focused on large components having > 5% of all reads, which in average account for 93.7% of all reads found in a patient. The distance between any two variants calculated over the component correlated strongly with nucleotide distances (r = 0.9499; p = 0.0001), a better correlation than the one obtained with Neighbour-Joining trees (r = 0.7624; p = 0.0001). In each patient, components were well separated, with the average distance between (6.53%) being 10 times greater than within each component (0.68%). The ratio of nonsynonymous to synonymous changes was calculated and some patients (6.9%) showed a mixture of networks under strong negative and positive selection. All components were robust to in silico stochastic sampling; even after randomly removing 85% of all reads, the largest connected component in the new subsample still involved 82.4% of remaining nodes. In vitro sampling showed that 93.02% of components present in the original sample were also found in experimental replicas, with 81.6% of reads found in both. When syringe-sharing transmission events were simulated, 91.2% of all simulated transmission events seeded all components present in the source. Conclusions: Most intra-host variants are organized into distinct single-mutation components that are: well separated from each other, represent genetic distances between viral variants, robust to sampling, reproducible and likely seeded during transmission events. Facilitated by NGS, large components offer a novel evolutionary framework for genetic analysis of intra-host viral populations and understanding transmission, immune escape and drug resistance

Effect of dietary docosahexaenoic acid supplementation on the participation of vasodilator factors in aorta from orchidectomized rats

Benefits of n-3 polyunsaturated fatty acids (PUFAs) against cardiovascular diseases have been reported. Vascular tone regulation is largely mediated by endothelial factors whose release is modulated by sex hormones. Since the incidence of cardiovascular pathologies has been correlated with decreased levels of sex hormones, the aim of this study was to analyze whether a diet supplemented with the specific PUFA docosahexaenoic acid (DHA) could prevent vascular changes induced by an impaired gonadal function. For this purpose, control and orchidectomized rats were fed with a standard diet supplemented with 5% (w/w) sunflower oil or with 3% (w/w) sunflower oil plus 2% (w/w) DHA. The lipid profile, the blood pressure, the production of prostanoids and nitric oxide (NO), and the redox status of biological samples from control and orchidectomized rats, fed control or DHA-supplemented diet, were analyzed. The vasodilator response and the contribution of NO, prostanoids and hyperpolarizing mechanisms were also studied. The results showed that orchidectomy negatively affected the lipid profile, increased the production of prostanoids and reactive oxygen species (ROS), and decreased NO production and the antioxidant capacity, as well as the participation of hyperpolarizing mechanisms in the vasodilator responses. The DHAsupplemented diet of the orchidectomized rats decreased the release of prostanoids and ROS, while increasing NO production and the antioxidant capacity, and it also improved the lipid profile. Additionally, it restored the participation of hyperpolarizing mechanisms by activating potassium. Since the modifications induced by the DHA-supplemented diet were observed in the orchidectomized, but not in the healthy group, DHA seems to exert cardioprotective effects in physiopathological situations in which vascular dysfunction existsThis study was supported by grants to MF from the Fondo de Investigaciones Sanitarias (PI1100406), Comunidad de Madrid (S2013/ABI- 2783, “INSPIRA1-CM”), Fondo Europeo de Desarrollo Regional, and Centro de Estudios América Latina (Grupo Santander-UAM)

Origen geno-geográfico de haplotipos STR del cromosoma Y en una muestra caucásico-mestiza y afrodescendiente de Colombia

Introduction: Y chromosome STR haplotypes have been widely used in population studies to establish the origin of diverse populations.Objective: We analyzed Y chromosome STR haplotypes (8 loci) in 134 Caucasian-mestizo and 137 African-descent Colombian unrelated individuals to correlate the geographical origin with historical data as well as the genetic relationships and possible admixture patterns.Materials and methods: One hundred samples of African descent and 137 Caucasian-mestizo samples analyzed for Y chromosome STR haplotypes by PCR followed by acrylamide electrophoresis.Results: No evidence of population substructure was found for the African descent. Two point fiftynine per cent of the haplotypes were shared between the two groups with the possible existence of Caucasian gene flow towards Afro-descendants.Conclusion: The Caucasian-Mestizo Colombian population is grouped with other populations of the Iberian Peninsula and Europe, while the Afro-Colombian population is grouped with other African populations reported. doi: http://dx.doi.org/10.7705/biomedica.v33i3.807 Introducción. Los haplotipos STR de cromosoma Y han sido ampliamente utilizados en estudios de poblaciones para establecer el origen de diversas poblaciones.Objetivo. Se analizaron haplotipos STR del cromosoma Y (8 loci) en 134 afrodescendientes y caucásico-mestizos no relacionados de Colombia, para correlacionar el origen geográfico con los datos históricos, así como las relaciones genéticas y posibles patrones de mezcla.Materiales y métodos. Se analizaron los haplotipos STR del cromosoma Y mediante PCR seguidas de electroforesis en acrilamida, de 134 muestras de afrodescendientes y 137 muestras de caucásicos mestizos.Resultados. No se encontró evidencia de subestructuración de la población afrodescendiente. El 2,59 % de los haplotipos eran compartidos en los dos grupos analizados, con la posible existencia deflujo génico de caucásico-mestizos hacia los afrodescendientes.Conclusión. La población caucásico-mestiza colombiana se agrupa con otras poblaciones de la península Ibérica y Europa, mientras que la población afrodescendiente colombiana se agrupa conotras poblaciones africanas reportadas. doi: http://dx.doi.org/10.7705/biomedica.v33i3.80

IND-Enabling Studies for a Clinical Trial to Genetically Program a Persistent Cancer-Targeted Immune System

PURPOSE: To improve persistence of adoptively transferred T-cell receptor (TCR)-engineered T cells and durable clinical responses, we designed a clinical trial to transplant genetically-modified hematopoietic stem cells (HSCs) together with adoptive cell transfer of T cells both engineered to express an NY-ESO-1 TCR. Here, we report the preclinical studies performed to enable an investigational new drug (IND) application. EXPERIMENTAL DESIGN: HSCs transduced with a lentiviral vector expressing NY-ESO-1 TCR and the PET reporter/suicide gene HSV1-sr39TK and T cells transduced with a retroviral vector expressing NY-ESO-1 TCR were coadministered to myelodepleted HLA-A2/Kb mice within a formal Good Laboratory Practice (GLP)-compliant study to demonstrate safety, persistence, and HSC differentiation into all blood lineages. Non-GLP experiments included assessment of transgene immunogenicity and in vitro viral insertion safety studies. Furthermore, Good Manufacturing Practice (GMP)-compliant cell production qualification runs were performed to establish the manufacturing protocols for clinical use. RESULTS: TCR genetically modified and ex vivo-cultured HSCs differentiated into all blood subsets in vivo after HSC transplantation, and coadministration of TCR-transduced T cells did not result in increased toxicity. The expression of NY-ESO-1 TCR and sr39TK transgenes did not have a detrimental effect on gene-modified HSC's differentiation to all blood cell lineages. There was no evidence of genotoxicity induced by the lentiviral vector. GMP batches of clinical-grade transgenic cells produced during qualification runs had adequate stability and functionality. CONCLUSIONS: Coadministration of HSCs and T cells expressing an NY-ESO-1 TCR is safe in preclinical models. The results presented in this article led to the FDA approval of IND 17471

Insight into genetic predisposition to chronic lymphocytic leukemia from integrative epigenomics.

Genome-wide association studies have provided evidence for inherited genetic predisposition to chronic lymphocytic leukemia (CLL). To gain insight into the mechanisms underlying CLL risk we analyze chromatin accessibility, active regulatory elements marked by H3K27ac, and DNA methylation at 42 risk loci in up to 486 primary CLLs. We identify that risk loci are significantly enriched for active chromatin in CLL with evidence of being CLL-specific or differentially regulated in normal B-cell development. We then use in situ promoter capture Hi-C, in conjunction with gene expression data to reveal likely target genes of the risk loci. Candidate target genes are enriched for pathways related to B-cell development such as MYC and BCL2 signalling. At 14 loci the analysis highlights 63 variants as the probable functional basis of CLL risk. By integrating genetic and epigenetic information our analysis reveals novel insights into the relationship between inherited predisposition and the regulatory chromatin landscape of CLL