Rananos expression pattern during oogenesis and early embryonic development in Rhynchosciara americana

The Dipteran a native Brazilian insect that has become a valuable model system for developmental biology research because it provides an interesting opportunity to study a different type of insect oogenesis. Sequences from a cDNA library that was constructed with poly A + RNA from the ovaries of larvae at different ages were analyzed. Molecular characterization confirmed interesting findings, such as the presence of . The gene encodes a conserved RNA-binding protein that is required during early development for the maintenance and division of the primordial germ cells of Diptera. plays an important role in specifying the posterior regions of insect embryos and is important for abdomen formation. In the present work, we showed the spatial and temporal expression profiles of this important gene, which is involved in oogenesis and early development. Data mining techniques were used to obtain the complete sequence of . Bioinformatic tools were used to determine the following: (1) the secondary structure of the 3'-untranslated region of the mRNA, (2) the encoded protein of the isolated gene, (3) the conserved zinc-finger domains of the Nanos protein, and (4) phylogenetic analyses. Furthermore, RNA in situ hybridization and immunolocalization were used to determine mRNA and protein expression in the tissues that were studied and to define as a germ cell molecular marker.

Long-term type 1 diabetes impairs decidualization and extracellular matrix remodeling during early embryonic development in mice

Introduction: Endometrial decidualization and associated extracellular matrix (ECM) remodeling are critical events to the establishment of the maternal-fetal interface and successful pregnancy. Here, we investigated the impact of type 1 diabetes on these processes during early embryonic development, in order to contribute to the understanding of the maternal factors associated to diabetic embryopathies. Methods: Alloxan-induced diabetic Swiss female mice were bred after different periods of time to determine the effects of diabetes progression on the development of gestational complications. Furthermore, the analyses focused on decidual development as well as mRNA expression, protein deposition and ultrastructural organization of decidual ECM. Results: Decreased number of implantation sites and decidual dimensions were observed in the group mated 90-110 days after diabetes induction (D), but not in the 50-70D group. Picrosirius staining showed augmentation in the fibrillar collagen network in the 90e110D group and, following immunohistochemical examination, that this was associated with increase in types I and V collagens and decrease in type III collagen and collagen-associated proteoglycans biglycan and lumican. qPCR, however, demonstrated that only type I collagen mRNA levels were increased in the diabetic group. Alterations in the molecular ratio among distinct collagen types and proteoglycans were associated with abnormal collagen fibrillogenesis, analyzed by transmission electron microscopy. Conclusions: Our results support the concept that the development of pregnancy complications is directly related with duration of diabetes (progression of the disease), and that this is a consequence of both systemic factors (i.e. disturbed maternal endocrine-metabolic profile) and uterine factors, including impaired decidualization and ECM remodeling

Heparin and penicillamine–hypotaurine–epinephrine (PHE) solution during bovine in vitro fertilization procedures impair the quality of spermatozoa but improve normal oocyte fecundation and early embryonic development

The presence of heparin and a mixture of penicillamine, hypotaurine, and epinephrine (PHE) solution in the in vitro fertilization (IVF) media seem to be a prerequisite when bovine spermatozoa are capacitated in vitro, in order to stimulate sperm motility and acrosome reaction. The present study was designed to determine the effect of the addition of heparin and PHE during IVF on the quality and penetrability of spermatozoa into bovine oocytes and on subsequent embryo development. Sperm quality, evaluated by the integrity of plasma and acrosomal membranes and mitochondrial function, was diminished (P<0.05) in the presence of heparin and PHE. Oocyte penetration and normal pronuclear formation rates, as well as the percentage of zygotes presenting more than two pronuclei, was higher (P<0.05) in the presence of heparin and PHE. No differences were observed in cleavage rates between treatment and control (P>0.05). However, the developmental rate to the blastocyst stage was increased in the presence of heparin and PHE (P>0.05). The quality of embryos that reached the blastocyst stage was evaluated by counting the inner cell mass (ICM) and trophectoderm (TE) cell numbers and total number of cells; the percentage of ICM and TE cells was unaffected (P>0.05) in the presence of heparin and PHE (P<0.05). In conclusion, this study demonstrated that while the supplementation of IVF media with heparin and PHE solution impairs spermatozoa quality, it plays an important role in sperm capacitation, improving pronuclear formation, and early embryonic development

Somatic Mosaic Activating Mutations in PIK3CA Cause CLOVES Syndrome

Congenital lipomatous overgrowth with vascular, epidermal, and skeletal anomalies (CLOVES) is a sporadically occurring, nonhereditary disorder characterized by asymmetric somatic hypertrophy and anomalies in multiple organs. We hypothesized that CLOVES syndrome would be caused by a somatic mutation arising during early embryonic development. Therefore, we employed massively parallel sequencing to search for somatic mosaic mutations in fresh, frozen, or fixed archival tissue from six affected individuals. We identified mutations in PIK3CA in all six individuals, and mutant allele frequencies ranged from 3% to 30% in affected tissue from multiple embryonic lineages. Interestingly, these same mutations have been identified in cancer cells, in which they increase phosphoinositide-3-kinase activity. We conclude that CLOVES is caused by postzygotic activating mutations in PIK3CA. The application of similar sequencing strategies will probably identify additional genetic causes for sporadically occurring, nonheritable malformations.

Absence of inactivating mutations and deletions in the DMRT1 and FGF9 genes in a large cohort of 46,XY patients with gonadal dysgenesis

Despite advances in our understanding of the mechanisms involved in sex determination and differentiation, the specific roles of many genes in these processes are not completely understood in humans. Both DMRT1 and FGF9 are among this group of genes. Dmrt1 controls germ cell differentiation, proliferation, migration and pluripotency and Sertoli cell proliferation and differentiation. Fgf9 has been considered a critical factor in early testicular development and germ cell survival in mice. We screened for the presence of DMRT1 and FGF9 mutations in 33 patients with 46,XY gonadal dysgenesis. No deletions in either DMRT1 or FGF9 were identified using the MLPA technique. Eight allelic variants of DMRT1 were identified, and in silico analysis suggested that the novel c.968-15insTTCTCTCT variant and the c.774G>C (rs146975077) variant could have potentially deleterious effects on the DMRT1 protein. Nine previously described FGF9 allelic variants and six different alleles of the 3' UTR microsatellite were identified. However, none of these DMRT1 or FGF9 variants was associated with increased 46,XY gonadal dysgenesis. In conclusion, our study suggests that neither DMRT1 nor FGF9 abnormalities are frequently involved in dysgenetic male gonad development in patients with non-syndromic 46,XY disorder of sex development. (C) 2012 Published by Elsevier Masson SAS.

Germline-Specific MATH-BTB Substrate Adaptor MAB1 Regulates Spindle Length and Nuclei Identity in Maize

Germline and early embryo development constitute ideal model systems to study the establishment of polarity, cell identity, and asymmetric cell divisions (ACDs) in plants. We describe here the function of the MATH-BTB domain protein MAB1 that is exclusively expressed in the germ lineages and the zygote of maize (Zea mays). mab1 (RNA interference [RNAi]) mutant plants display chromosome segregation defects and short spindles during meiosis that cause insufficient separation and migration of nuclei. After the meiosis-to-mitosis transition, two attached nuclei of similar identity are formed in mab1 (RNAi) mutants leading to an arrest of further germline development. Transient expression studies of MAB1 in tobacco (Nicotiana tabacum) Bright Yellow-2 cells revealed a cell cycle-dependent nuclear localization pattern but no direct colocalization with the spindle apparatus. MAB1 is able to form homodimers and interacts with the E3 ubiquitin ligase component Cullin 3a (CUL3a) in the cytoplasm, likely as a substrate-specific adapter protein. The microtubule-severing subunit p60 of katanin was identified as a candidate substrate for MAB1, suggesting that MAB1 resembles the animal key ACD regulator Maternal Effect Lethal 26 (MEL-26). In summary, our findings provide further evidence for the importance of posttranslational regulation for asymmetric divisions and germline progression in plants and identified an unstable key protein that seems to be involved in regulating the stability of a spindle apparatus regulator(s).

Plant invasions research in Latin America: fast track to a more focused agenda

While many developed countries have invested heavily in research on plant invasions over the last 50 years, the immense region of Latin America has made little progress. Recognising this, a group of scientists working on plant invasions in Latin America met in Chile in late 2010 to develop a research agenda for the region based on lessons learned elsewhere. Our three main findings are as follows. (1) Globalisation is inevitable, but the resultant plant introductions can be slowed or prevented by effective quarantine and early intervention. Development of spatially explicit inventories, research on the invasion process and weed risk assessments can help prioritise and streamline action. (2) Eradication has limited application for plants and control is expensive and requires strict prioritisation and careful planning and evaluation. (3) Accepting the concept of novel ecosystems, new combinations of native and introduced species that no longer depend on human intervention, may help optimise invasive species management. Our vision of novel ecosystem management is through actions that: (a) maintain as much native biodiversity and ecosystem functionality as possible, (b) minimise management intervention to invasives with known impact, and (c) maximise the area of intervention. We propose the creation of a Latin American Invasive Plants Network to help focus the new research agenda for member countries. The network would coordinate research and training and establish funding priorities, develop and strengthen tools to share knowledge, and raise awareness at the community, governmental and intergovernmental levels about the social, economic and environmental costs of plant invasions.

Perfil linguístico de crianças com alteração específica de linguagem

OBJETIVO: Caracterizar o perfil linguístico de crianças com alteração específica de linguagem (AEL) utilizando a versão brasileira do Test of Early Language Development - 3rd edition (TELD-3); comparar as idades cronológica e linguística, e classificar a severidade do quadro. MÉTODOS: O teste foi aplicado individualmente a 46 crianças com idades entre 2 anos e 10 meses e 7 anos e 11 meses, diagnosticadas com AEL, que estavam em atendimento fonoaudiológico semanal. A partir dos dados obtidos, foi realizada a comparação entre a média da idade cronológica e a média da idade linguística equivalente. O tipo de comprometimento foi classificado em misto ou puramente expressivo e o grau de severidade foi estabelecido. RESULTADOS: O comprometimento misto foi o mais frequente nas crianças com AEL, porém a classificação da severidade indicou que a categoria leve foi a mais frequente, tanto na recepção quanto na expressão. A idade linguística esteve abaixo da idade cronológica na maioria dos sujeitos, em ambos os subtestes. A linguagem expressiva foi a mais prejudicada, visto que os sujeitos apresentaram menor média de idade linguística equivalente, além de ter havido maior concentração de sujeitos classificados com alteração abaixo da média e com gravidade mais acentuada. CONCLUSÃO: Nesta população predominam os quadros mistos, com maior prejuízo da expressão e cuja severidade é considerada leve. Além disso, o TELD-3 mostrou ser um instrumento útil no processo diagnóstico destas alterações de linguagem.

Pain in preterm infants: effects of sex, gestational age, and neonatal illness severity

Neonates hospitalized in a neonatal intensive care unit are exposed to many painful and stressful procedures. Biobehavioral pain reactivity in preterm infants during the neonatal period may reflect the capacity of the central nervous system to regulate arousal and neurobiological organization. We review empirical studies on the effects of sex, gestational age, and neonatal illness severity on pain reactivity in children born preterm. A literature search was conducted using PubMed, Institute of Scientific Information Web of Science, PsycINFO, Latin American and Caribbean Health Sciences Literature, and Scientific Electronic Library Online databases. Additionally, a special search was performed in online journals that publish pain studies including Pain, Early Human Development, European Journal of Pain, and Pain Management Nursing. The literature search covered the period from 2004 to 2009. Data were extracted according to predefined inclusion and exclusion criteria. Of the 18 studies reviewed, 16 analyzed gestational age, 13 examined neonatal illness severity, and eight focused on sex. Most of the studies analyzed more than one of these three variables. The majority of the studies found effects of gestational age (n = 14) and neonatal illness severity (n = 11) on pain responses. Only two studies found an influence of sex on infant pain responses. In conclusion, gestational age and neonatal illness severity influence pain responses in infants born preterm. Further studies should be conducted to examine the influence of sex on pain responses.

Early development, survival and growth rates of the giant clam Tridacna crocea (Bivalvia: Tridacnidae)

Tridacnid clams are conspicuous inhabitants of Indo-Pacific coral reefs and are traded and cultivated for the aquarium and food industries. In the present study, daily growth rates of larvae of the giant clam Tridacna crocea were determined in the laboratory during the first week of life. Adults were induced to spawn via intra-gonadal serotonin injection through the byssal orifice. After spawning oocytes were collected, fertilized and kept in 3 L glass beakers and raceways treated with antibiotics to avoid culture contamination. Larvae were fed twice with the microalga Isochrysis galbana and zooxanthellae were also offered twice during the veliger stage (days 4 and 6). Larval length was measured using a digitizing tablet coupled to a microcomputer. Larval mortality was exponential during the first 48 hours of life declining significantly afterwards. Mean growth rate was 11.3 mu m day-1, increasing after addition of symbionts to 18.0 mu m day-1. Survival increased to ca. 75% after the addition of zooxanthellae. The results describe the growth curve for T. crocea larvae and suggest that the acquisition of symbionts by larvae may be useful for larval growth and survival even before larvae have attained metamorphosis.

Fatty acids as a tool to compare cachara (Pseudoplatystoma reticulatum) (Siluriformes: Pimelodidae) and hybrid (Pseudoplatystoma corruscans x Pseudoplatystoma reticulatum) larvae during early development

In this study, we investigated the physiological alterations during ontogeny for cachara (Pseudoplatystoma reticulatum) and their hybrid larvae (Pseudoplatystoma corruscans x P. reticulatum) using lipids and fatty acids as physiological tools to elucidate the basis for differences in these groups' productivity in an industrial setting. Eggs and larvae samples were collected during January and February of 2008 in the city of Bandeirantes, MS, and were divided into three primary phases: phase I (0-16 h after fertilization); phase II (24 h after fertilization to 6 days after fertilization); and phase III (7-25 days after fertilization). The larvae of both groups showed a high degree of similarity, suggesting that the hybrid larvae showed a high level of heritability from the cachara broodstock. Analysis of the total lipid content provided evidence that there is no alteration in lipid concentration during ontogeny for both groups (i.e., the cachara and hybrids). However, the fatty acid profile showed that during the endogenous feeding period (phase II), when the larvae must use the energy reserves from the mother, the cachara larvae used mainly monounsaturated fatty acids for development. This is typical for most fish species, though notably, the hybrids preferentially used saturated fatty acids. Furthermore, certain specific changes demonstrate unique patterns of energy utilization and structural substrates, which may aid in elucidating the empirical differences reported by fish farmers (i.e., that the hybrids perform better than cacharas in captivity).

Early development, survival and growth rates of the giant clam Tridacna crocea (Bivalvia: Tridacnidae)

Tridacnid clams are conspicuous inhabitants of Indo-Pacific coral reefs and are traded and cultivated for the aquarium and food industries. In the present study, daily growth rates of larvae of the giant clam Tridacna crocea were determined in the laboratory during the first week of life. Adults were induced to spawn via intra-gonadal serotonin injection through the byssal orifice. After spawning oocytes were collected, fertilized and kept in 3 L glass beakers and raceways treated with antibiotics to avoid culture contamination. Larvae were fed twice with the microalga Isochrysis galbana and zooxanthellae were also offered twice during the veliger stage (days 4 and 6). Larval length was measured using a digitizing tablet coupled to a microcomputer. Larval mortality was exponential during the first 48 hours of life declining significantly afterwards. Mean growth rate was 11.3 μm day-1, increasing after addition of symbionts to 18.0 μm day-1. Survival increased to ca. 75% after the addition of zooxanthellae. The results describe the growth curve for T. crocea larvae and suggest that the acquisition of symbionts by larvae may be useful for larval growth and survival even before larvae have attained metamorphosis.