DCs Pulsed with Novel HLA-A2-Restricted CTL Epitopes against Hepatitis C Virus Induced a Broadly Reactive Anti-HCV-Specific T Lymphocyte Response

OBJECTIVE: To determine the capacity of dendritic cells (DCs) loaded with single or multiple-peptide mixtures of novel hepatitis C virus (HCV) epitopes to stimulate HCV-specific cytotoxic T lymphocyte (CTL) effector functions. METHODS: A bioinformatics approach was used to predict HLA-A2-restricted HCV-specific CTL epitopes, and the predicted peptides identified from this screen were synthesized. Subsequent IFN-γ ELISPOT analysis detected the stimulating function of these peptides in peripheral blood mononuclear cells (PBMCs) from both chronic and self-limited HCV infected subjects (subjects exhibiting spontaneous HCV clearance). Mature DCs, derived in vitro from CD14(+) monocytes harvested from the study subjects by incubation with appropriate cytokine cocktails, were loaded with novel peptide or epitope peptide mixtures and co-cultured with autologous T lymphocytes. Granzyme B (GrB) and IFN-γ ELISPOT analysis was used to test for epitope-specific CTL responses. T-cell-derived cytokines contained in the co-cultured supernatant were detected by flow cytometry. RESULTS: We identified 7 novel HLA-A2-restricted HCV-specific CTL epitopes that increased the frequency of IFN-γ-producing T cells compared to other epitopes, as assayed by measuring spot forming cells (SFCs). Two epitopes had the strongest stimulating capability in the self-limited subjects, one found in the E2 and one in the NS2 region of HCV; five epitopes had a strong stimulating capacity in both chronic and self-limited HCV infection, but were stronger in the self-limited subjects. They were distributed in E2, NS2, NS3, NS4, and NS5 regions of HCV, respectively. We also found that mDCs loaded with novel peptide mixtures could significantly increase GrB and IFN-γ SFCs as compared to single peptides, especially in chronic HCV infection subjects. Additionally, we found that DCs pulsed with multiple epitope peptide mixtures induced a Th1-biased immune response. CONCLUSIONS: Seven novel and strongly stimulating HLA-A2-restricted HCV-specific CTL epitopes were identified. Furthermore, DCs loaded with multiple-epitope peptide mixtures induced epitope-specific CTLs responses

A importância da associação obesidade e gravidez

Characteristics of the evolution of pregnancy in obese women were studied for their effect on newborn infants. Two control groups were observed - one of normal weight pregnant women, one of obese. The variables selected were: the socio-economic status of the family and the mother's age, height, arm circunference, prepregancy weight, total number of pregnancies, parity, weight gain during pregnancy, obstetric complications, birth weight, and fetal vitality. Results showed that pregnancy in obese women differs from that in normal weight women and that they show a larger incidence of obstetric complications. Children of obese mothers had a higher mortality rate principally in the perinatal period; moreover, there was also a higher rate of prematurity and a higher proportion of overweight babies among obese mothers. As a result, the distribution of the curve of the birth weight of infants of obese morthers was higher than that of infants of normal weight mothers. The conclusion reached was that whenever a pregnant obese woman reduced foot intake, with resultant insufficient weight gain, intrauterine growth was affected. Thus, it follows that pregnancy is not the best time for the obese mother to lose weight; for this reason, it is important that she receive adequate guidance in regard to diet. Obesity, therefore, is a factor contributing to high-risk pregnancy which can affect both mother and child.Foram estudados dois grupos de gestantes, sendo um de grávidas normais e outro de obesas, com a finalidade de reconhecer algumas características da evolução da gravidez, em mulheres obesas, e suas repercussões sobre o concepto. Foram relacionadas as seguintes variáveis: status sócio-econômico familiar, idade, altura, perímetro braquial, peso habitual, número de gestações anteriores, paridade materna, ganho de peso durante a gestação, idade gestacional, intercorrências durante a gestação, peso ao nascer e vitalidade do recém-nascido. Pelos resultados concluiu-se que as gestantes obesas são diferentes das normais e apresentam maior incidência de complicações obstétricas. Os recém-nascidos, filhos de obesas, registraram índice maior de mortalidade, principalmente no período perinatal. Houve maior incidência de prematuridade e de fetos macrossômicos, sendo a curva de distribuição de peso ao nascer diferente da dos recém-nascidos das gestantes normais. A média de peso ao nascer das crianças das gestantes obesas é maior que o das normais. Concluiu-se ainda que toda vez que a gestante obesa sofre restrição alimentar, com ganho de peso inadequado, o crescimento intra-uterino é afetado; não sendo, portanto, a época da gravidez a melhor para a obesa perder peso, mas, ao contrário, ela deveria receber uma orientação alimentar adequada. A obesidade é pois um fator de aumento do risco gravídico, que pode afetar tanto a mãe como o concepto

Genome-Wide Analyses of Exonic Copy Number Variants in a Family-Based Study Point to Novel Autism Susceptibility Genes

The genetics underlying the autism spectrum disorders (ASDs) is complex and remains poorly understood. Previous work has demonstrated an important role for structural variation in a subset of cases, but has lacked the resolution necessary to move beyond detection of large regions of potential interest to identification of individual genes. To pinpoint genes likely to contribute to ASD etiology, we performed high density genotyping in 912 multiplex families from the Autism Genetics Resource Exchange (AGRE) collection and contrasted results to those obtained for 1,488 healthy controls. Through prioritization of exonic deletions (eDels), exonic duplications (eDups), and whole gene duplication events (gDups), we identified more than 150 loci harboring rare variants in multiple unrelated probands, but no controls. Importantly, 27 of these were confirmed on examination of an independent replication cohort comprised of 859 cases and an additional 1,051 controls. Rare variants at known loci, including exonic deletions at NRXN1 and whole gene duplications encompassing UBE3A and several other genes in the 15q11–q13 region, were observed in the course of these analyses. Strong support was likewise observed for previously unreported genes such as BZRAP1, an adaptor molecule known to regulate synaptic transmission, with eDels or eDups observed in twelve unrelated cases but no controls (p = 2.3×10−5). Less is known about MDGA2, likewise observed to be case-specific (p = 1.3×10−4). But, it is notable that the encoded protein shows an unexpectedly high similarity to Contactin 4 (BLAST E-value = 3×10−39), which has also been linked to disease. That hundreds of distinct rare variants were each seen only once further highlights complexity in the ASDs and points to the continued need for larger cohorts

Improved Induced-Spawning Protocol for the Spotted Rose Snapper (Lutjanus guttatus)

An improved protocol for hormonal induction of spawning in spotted rose snapper (Lutjanus guttatus) resulted in increased spawning efficiency in newly-caught wild breeders and wild-caught adults maintained in captivity for more than a year. A controlled-release delivery system (implant) based on an ethylene-vinyl acetate copolymer (EVAc) matrix was loaded with gonadotropin-releasing hormone agonist (GnRHa). The required GnRHa dose was established in two stages. The first stage included meta-analysis of our earlier experiments with wild spawners; the second stage included new experiments with wild and captive breeders. A nomograph was devel- oped to calculate the required GnRHa implant dose, taking into account the origin of the female (wild vs. captive), the initial mean oocyte diameter, and body weight. The effective GnRHa dose was greater in wild than captive females and, in both cases, inversely related to mean oocyte diameter. Using this nomograph, over half the wild females with a mean oocyte diam- eter of ≥425 μm and over half the captive females with a mean oocyte diam- eter of ≥350 μm responded to GnRHa implant treatment (producing multi- ple spawning events in captives), with mean total relative fecundity ranging 80-278 x 103 eggs/kg body weight and 51-85% fertilization success. The nomograph can be used to calculate the GnRHa implant dose required to induce spawning in this species under commercial aquaculture conditions

Mugil curema and M. liza Valenciennes in Cuvier & Valenciennes, 1836 (Osteichthyes, Perciformes): proposed conservation of the specific names

Volume: 49Start Page: 271End Page: 27

Studies in egg handling and incubation in the spotted rose snapper Lutjanus guttatus (Pisces Lutjanidae) [Estudios sobre el manejo e incubaci�n de huevos del pargo flamenco Lutjanus guttatus (Pisces, Lutjanidae)]

Egg handling and incubation are short but important procedures in juvenile fish production technologies. To determine appropriate practices for egg handling and incubation in the spotted rose snapper Lutjanus guttatus (Steindachner, 1969), three experiments were carried out in 100-L fiberglass conical cylinder tanks. Eggs were incubated with and without water flow (30% volume/h), at two densities (250 eggs/L and 1000 eggs/L), and with and without formaldehyde 10-ppm prophylactic treatment for 1-h prior to incubation. Total hatching and larvae survival percentages were not significantly different between the treatments with and without water flow, while results for larvae survival at hatching time and 48 h post-hatch stage were significantly better without water flow. For incubation at two different densities, all examined performance indexes were significantly better at the lower density, except for larvae survival at 48 h after hatching. The eggs with and without prophylactic treatment were not significantly different in all analyzed survival indexes at hatching time and 48 h post-hatch. Total length was significantly different in all treatments at hatching time and 48 h post-hatch, except in the incubation without water flow. The longest larvae were obtained in the experiment with prophylactic treatment

Studies in egg handling and incubation in the spotted rose snapper Lutjanus guttatus (Pisces Lutjanidae) [Estudios sobre el manejo e incubación de huevos del pargo flamenco Lutjanus guttatus (Pisces, Lutjanidae)]

Egg handling and incubation are short but important procedures in juvenile fish production technologies. To determine appropriate practices for egg handling and incubation in the spotted rose snapper Lutjanus guttatus (Steindachner, 1969), three experiments were carried out in 100-L fiberglass conical cylinder tanks. Eggs were incubated with and without water flow (30% volume/h), at two densities (250 eggs/L and 1000 eggs/L), and with and without formaldehyde 10-ppm prophylactic treatment for 1-h prior to incubation. Total hatching and larvae survival percentages were not significantly different between the treatments with and without water flow, while results for larvae survival at hatching time and 48 h post-hatch stage were significantly better without water flow. For incubation at two different densities, all examined performance indexes were significantly better at the lower density, except for larvae survival at 48 h after hatching. The eggs with and without prophylactic treatment were not significantly different in all analyzed survival indexes at hatching time and 48 h post-hatch. Total length was significantly different in all treatments at hatching time and 48 h post-hatch, except in the incubation without water flow. The longest larvae were obtained in the experiment with prophylactic treatment

Production of Juveniles of the mullet Mugil liza Valenciennes, 1836, by controlled reproduction in Cuba

Induced spawning and larval rearing experiments were carried out with the mullet Mugil liza Valenciennes at the end of the 1987/1988 natural spawning season at Tunas de Zaza, in which the juvenile stage of a  mugilid species was reached for the first time in Cuba. All examined females had atretic oocytes. From the selected 18, four (22.2%) died during the hormone treatment and from the 14 that finished it, natural spawning or artificial fertilization was obtained from seven of them (50%). Best spawning results were obtained with 60-90 mg/kg of mullet pituitary in 3-4 partial doses (24 h interval). The eggs of one female with the highest fertilization rate (60%) were incubated at 24ºC, obtaining 90% survival of the fertilized eggs at hatching. The rearing experiments demonstrated that larvae were ready for exogenous feeding at day 3-4 and that the postlarval stage ended 40 days after hatching; also, that tarpaulin tanks were not adequate, that a strong aeration at the centre is beneficial and that the use of an intermediate food organism between rotifers and Artemia should improve the results. At the end of the rearing cycle (49 days), 220 juveniles were harvested in one of the four treatments used (0.20% survival). The difficulties faced and possible technique improvements for increasing survival are discussed