Stimulation of multiple mitogen-activated protein kinase sub-families by oxidative stress and phosphorylation of the small heat shock protein, HSP25/27, in neonatal ventricular myocytes.

We investigated the activation of three subfamilies of mitogen-activated protein kinases (MAPKs), namely the stress-activated protein kinases/c-Jun N-terminal kinases (SAPKs/JNKs), the extracellularly responsive kinases (ERKs) and p38-MAPK, by oxidative stress as exemplified by H2O2 in primary cultures of neonatal rat ventricular myocytes. The 46 and 54 kDa species of SAPKs/JNKs were activated 5- and 10-fold, respectively, by 0.1 mM H2O2 (the maximally effective concentration). Maximal activation occurred at 15-30 min, but was still detectable after 2 h. Both ERK1 and ERK2 were activated 16-fold by 0.1 mM H2O2 with a similar time course to the SAPKs/JNKs, and this was comparable with their activation by 1 microM PMA, the most powerful activator of ERKs that we have so far identified in these cells. The activation of ERKs by H2O2 was inhibited by PD98059, which inhibits the activation of MAPK (or ERK) kinases, and by the protein kinase C (PKC) inhibitor, GF109203X. ERK activation was also inhibited by down-regulation of PMA-sensitive PKC isoforms. p38-MAPK was activated by 0.1 mM H2O2 as shown by an increase in its phosphorylation. However, maximal phosphorylation (activation) was more rapid (<5 min) than for the SAPKs/JNKs or the ERKs. We studied the downstream consequences of p38-MAPK activation by examining activation of MAPK-activated protein kinase 2 (MAPKAPK2) and phosphorylation of the MAPKAPK2 substrate, the small heat shock protein HSP25/27. As with p38-MAPK, MAPKAPK2 was rapidly activated (maximal within 5 min) by 0.1 mM H2O2. This activation was abolished by 10 microM SB203580, a selective inhibitor of certain p38-MAPK isoforms. The phosphorylation of HSP25/27 rapidly followed activation of MAPKAPK2 and was also inhibited by SB203580. Phosphorylation of HSP25/27 was associated with a decrease in its aggregation state. These data indicate that oxidative stress is a powerful activator of all three MAPK subfamilies in neonatal rat ventricular myocytes. Activation of all three MAPKs has been associated with the development of the hypertrophic phenotype. However, stimulation of p38-MAPK and the consequent phosphorylation of HSP25/27 may also be important in cardioprotection.

"When I go to bed hungry and sleep, I'm not hungry": children and parents' experiences of food insecurity

Evidence demonstrates food insecurity has a detrimental impact on a range of outcomes for children, but little research has been conducted in the UK, and children have rarely been asked to describe their experiences directly. We examined the experiences of food insecure families living in South London. Our mixed-methods approach comprised a survey of parents (n = 72) and one-to-one semi-structured interviews with children aged 5-11 years (n = 19). The majority of parents (86%) described their food security during the preceding year as very low. Most reported they had often or sometimes had insufficient food, and almost all had worried about running out of food. Two thirds of parents had gone hungry. Most parents reported they had been unable to afford a nutritionally balanced diet for their children, and just under half reported that their children had gone hungry. Four themes emerged from the interviews with children: sources of food; security of food, nutritional quality of food, and experiences of hunger. Children's descriptions of insufficient food being available indicate that parents are not always able to shield them from the impact of food insecurity. The lack of school-meals and after-school clubs serving food made weekends particularly problematic for some children. A notable consequence of food insecurity appears to be reliance on low-cost takeaway food, likely to be nutritionally poor.

Conflicting language ideologies and contradictory language practices in Singaporean bilingual families

Informed by family language policy (FLP) as the theoretical framework, I illustrate in this paper how language ideologies can be incongruous and language policies can be conflicting through three multilingual families in Singapore representing three major ethnic groups – Chinese, Malay and Indian. By studying their family language audits, observing their language practices, and engaging in conversations about their language ideologies, I look at what these families do and do not do and what they claim to do and not to do. Data were collected over a period of 6 months with more than 700 minutes of recording of actual interactions. Analysis of the data reveals that language ideologies are ‘power-inflected’ and tend to become the source of educational and social tensions which in turn shape family language practices. In Singapore these tensions are illustrated by the bilingual policy recognising mother tongues (MTs) and English as official languages, and its educational policy establishing English as the medium of instruction. The view of English as having instrumental values and MTs as having cultural functions reveals that language choices and practices in family domains are value-laden in everyday interactions and explicitly negotiated and established through FLP.

Estrogen regulation of chicken riboflavin carrier protein gene is mediated by ERE half sites without direct binding of estrogen receptor

Estrogen is an important steroid hormone that mediates most of its effects on regulation of gene expression by binding to intracellular receptors. The consensus estrogen response element (ERE) is a 13 bp palindromic inverted repeat with a three nucleotide spacer. However, several reports suggest that many estrogen target genes are regulated by diverse elements, such as imperfect EREs and ERE half sites (ERE 1/2), which are either the proximal or the distal half of the palindrome. To gain more insight into ERE half site-mediated gene regulation, we used a region from the estrogen-regulated chicken riboflavin carrier protein (RCP) gene promoter that contains ERE half sites. Using moxestrol, an analogue of estrogen and transient transfection of deletion and mutation containing RCP promoter/reporter constructs in chicken hepatoma (LMH2A) cells, we identified an estrogen response unit (ERU) composed of two consensus ERE 1/2 sites and one non-consensus ERE 1/2 site. Mutation of any of these sites within this ERU abolishes moxestrol response. Further, the ERU is able to confer moxestrol responsiveness to a heterologous promoter. Interestingly, RCP promoter is regulated by moxestrol in estrogen responsive human MCF-7 cells, but not in other cell lines such as NIH3T3 and HepG2 despite estrogen receptor-alpha (ER-�) co transfection. Electrophoretic mobility shift assays (EMSAs) with promoter regions encompassing the half sites and nuclear extracts from LMH2A cells show the presence of a moxestrol-induced complex that is abolished by a polyclonal anti-ER� antibody. Surprisingly, estrogen receptor cannot bind to these promoter elements in isolation. Thus, there appears to be a definite requirement for some other factor(s) in addition to estrogen receptor, for the generation of a suitable response of this promoter to estrogen. Our studies therefore suggest a novel mechanism of gene regulation by estrogen, involving ERE half sites without direct binding of ER to the cognate elements.

Clinical evaluation and COL2A1 gene analysis in 21 Brazilian families with Stickler syndrome: Identification of novel mutations, further genotype/phenotype correlation, and its implications for the diagnosis

We present clinical and molecular evaluation from a large cohort of patients with Stickler syndrome: 78 individuals from 21 unrelated Brazilian families. The patients were selected in a Hospital with a craniofacial dysmorphology assistance service and clinical diagnosis was based on the presence of cleft palate associated to facial and ocular anomalies of Stickler syndrome. Analysis of COL2A1 gene revealed 9 novel and 4 previously described pathogenic mutations. Except for the mutation c.556G>T (p.Gly186X), all the others were located in the triple helical domain. We did not find genotype/phenotype correlation in relation to type and position of the mutation in the triple helical domain. However, a significantly higher proportion of myopia in patients with mutations located in this domain was observed in relation to those with the mutation in the non-tripe helical domain (c.556G>T; P < 0.04). A trend towards a higher prevalence of glaucoma, although not statistically significant, was observed in the presence of the mutation c.556G>T. It is possible. that this mutation alters the splicing of the mRNA instead of only creating a premature stop codon and therefore it can lead to protein products of different ocular effects. One novel DNA variation (c.1266+7G>C) occurs near a splice site and it was observed to co-segregate with the phenotype in one of the two families with this DNA variation. As in silico analysis predicted that the c.1266+7G>C DNA variation can affect the efficiency of the splicing, we still cannot rule it out as non-pathogenic. Our study also showed that ascertainment through cleft palate associated to other craniofacial signs can be very efficient for identification of Stickler syndrome patients. Still, high frequency of familial cases and high frequency of underdevelopment of distal lateral tibial epiphyses observed in our patients suggested that the inclusion of this information can improve the clinical diagnosis of Stickler syndrome. (C) 2008 Elsevier Masson SAS. All rights reserved.

Prevalence and nonrandom distribution of exonic mutations in interferon regulatory factor 6 in 307 families with Van der Woude syndrome and 37 families with popliteal pterygium syndrome

Purpose: Interferon regulatory factor 6 encodes a member of the IRF family of transcription factors. Mutations in interferon regulatory factor 6 cause Van der Woude and popliteal pterygium syndrome, two related orofacial clefting disorders. Here, we compared and contrasted the frequency and distribution of exonic Mutations in interferon regulatory factor 6 between two large geographically distinct collections of families with Van der Woude and between one collection of families with popliteal pterygium syndrome. Methods: We performed direct sequence analysis of interferon regulatory factor 6 exons oil samples from three collections, two with Van der Woude and one with popliteal pterygium syndrome. Results: We identified mutations in interferon regulatory factor 6 exons in 68% of families in both Van der Woude collections and in 97% of families with popliteal pterygium syndrome. In sum, 106 novel disease-causing variants were found. The distribution of mutations in the interferon regulatory factor 6 exons in each collection was not random; exons 3, 4, 7, and 9 accounted for 80%. In the Van der Woude collections, the mutations were evenly divided between protein truncation and missense, whereas most mutations identified in the popliteal pterygium syndrome collection were missense. Further, the missense mutations associated with popliteal pterygium syndrome were localized significantly to exon 4, at residues that are predicted to bind directly to DNA. Conclusion: The nonrandom distribution of mutations in the interferon regulatory factor 6 exons suggests a two-tier approach for efficient mutation screens for interferon regulatory factor 6. The type and distribution of mutations are consistent with the hypothesis that Van der Woude is caused by haploinsufficiency of interferon regulatory factor 6. Oil the other hand, the distribution of popliteal pterygium syndrome-associated mutations suggests a different, though not mutually exclusive, effect oil interferon regulatory factor 6 function. Genet Med 2009:11(4):241-247.

Mobius Sequence in a Girl and Arthrogryposis in her Half-Brother: Distinct Phenotypes Caused By Prenatal Injuries

Mobius sequence is a congenital facial and abducens nerve palsy, frequently associated to abnormalities of extremities. Arthrogryposis multiplex congenital is defined as a congenital fixation of multiple joints seldom of neurogenic origin. Both sequences must have a genetic origin, but usually are sporadic cases related to environmental factors such as drugs exposition and maternal trauma. A 5-year-old girl and a 1-year-old boy were born with Mobius sequence and arthrogryposis multiplex congenital, respectively. During pregnancies, the mother had vaginal bleeding at 7 weeks and used crack (free-based cocaine) in the first trimester, respectively. The girl also has equinovarus talipes and autistic behavior. The boy has arthrogryposis with flexion contractures of the feet and knees. A vascular disruption, due to hemorrhage and cocaine exposure, causing a transient ischemic insult to embryos in a critical period of development may be responsible for distinct phenotypes in these cases.

HTR1B and HTR2C in autism spectrum disorders in Brazilian families

Autism spectrum disorders (ASD) is a group of behaviorally defined neuro developmental disabilities characterized by multiple genetic etiologies and a complex presentation. Several studies suggest the involvement of the serotonin system in the development of ASD, but only few have investigated serotonin receptors. We have performed a case-control and a family-based study with 9 polymorphisms mapped to two serotonin receptor genes (HTR1B and HTR2C) in 252 Brazilian male ASD patients of European ancestry. These analyses showed evidence of undertransmission of the HTR1B haplotypes containing alleles -161G and -261A at HTR1B gene to ASD (P=0.003), but no involvement of HTR2C to the predisposition to this disease. Considering the relatively low level of statistical significance and the power of our sample, further studies are required to confirm the association of these serotonin-related genes and ASD. (C) 2008 Elsevier B.V. All rights reserved.

Creatine supplementation reduces plasma levels of pro-inflammatory cytokines and PGE(2) after a half-ironman competition

Objective. The effect of creatine supplementation upon plasma levels of pro-inflammatory cytokines: Interleukin (IL) 1 beta and IL-6, Tumor Necrosis Factor alpha (TNF alpha), and Interferon alpha (INF alpha) and Prostaglandin E(2) (PGE(2)) after a half-ironman competition were investigated. Methods. Eleven triathletes, each with at least three years experience of participation in this sport were randomly divided between the control and experimental groups. During 5 days prior to competition, the control group (n = 6) was supplemented with carbohydrate (20g center dot d(-1)) whereas the experimental group (n = 5) received creatine (20 center dot d(-1)) in a double-blind trial. Blood samples were collected 48h before and 24 and 48h after competition and were used for the measurement of cytokines and PGE(2). Results. Forty-eight hours prior to competition there was no difference between groups in the plasma concentrations (pg center dot ml(-1), mean +/- SEM) of IL-6 (7.08 +/- 0.63), TNF alpha (76.50 +/- 5.60), INF alpha (18.32 +/- 1.20), IL-1 beta (23.42 +/- 5.52), and PGE(2) (39.71 +/- 3.8). Twenty-four and 48h after competition plasma levels of TNF alpha, INF alpha, IL-1 beta and PGE(2) were significantly increased (P < 0.05) in both groups. However, the increases in these were markedly reduced following creatine supplementation. An increase in plasma IL-6 was observed only after 24h and, in this case, there was no difference between the two groups. Conclusion. Creatine supplementation before a long distance triathlon competition may reduce the inflammatory response induced by this form of strenuous of exercise.

The network of intertextual relations in Naipaul's half a life and magic seeds

Esta dissertação mapeia a rede de relações intertextuais em Half a Life (2001) e sua continuação Magic Seeds (2004), os romances mais recentes do Prêmio Nobel de Literatura de 2001, V. S. Naipaul, como contribuição para o estudo da obra do autor. A noção de intertextualidade permeia os estudos literários, e o termo tem sido largamente empregado desde que foi cunhado por Julia Kristeva nos anos sessenta. Desde então as mais variadas, e muitas vezes divergentes, teorias sobre intertextualidade compartilham a idéia de que um texto só adquire significado pleno na interação com outros textos. A abordagem metodológica proposta é baseada na teoria da transtextualidade de Gérard Genette. Esta escolha implica o estudo de intertextos, paratextos, metatextos, arquitextos e hipertextos que constituem a interface entre os dois romances e outros escritos. O nome do protagonista "William Somerset Chandran" constitui o fio que guia o estudo das várias relações transtextuais nos dois romances. A partir do prenome do protagonista – William – este estudo situa os romances no contexto da tradição do Bildungsroman, e argumenta que estes estabelecem uma paródia arquitextual do gênero na medida em que subvertem seu cerne, ou seja, a formação do caráter do protagonista. O nome do meio do protagonista – Somerset – remete à ficcionalização do escritor Somerset Maugham na narrativa, ao mesmo tempo em que esta desmistifica a ótica ocidental sobre o hinduísmo popularizada por Maugham em The Razor's Edge. O sobrenome do protagonista – Chandran – leva ao estudo do conjunto de referências à origem indiana de Naipaul e o papel desta na produção do autor. Este nome se reporta ao romance de Narayan The Bachelor of Arts, cujo protagonista também é nomeado Chandran. Narayan é um escritor de destaque na literatura anglo-indiana e referência recorrente na obra de Naipaul. Os temas de migração e choque cultural apresentados nos dois romances têm sido presença constante na obra de Naipaul. Esta pesquisa mapeia a relação de continuidade entre os dois romances em questão e o conjunto da obra de Naipaul, salientando o papel da ambientação geográfica da narrativa, marcada pela jornada do protagonista através de três continentes. A teoria da transtextualidade é uma ferramenta operacional para a pesquisa, a qual examina a densidade das referências geográficas, históricas e literárias em Half a Life e Magic Seeds, visando aportar elementos para o estudo da produção literária de Naipaul, na medida em que estes romances recentes condensam e revisitam a visão de mundo deste autor.

"Não somos diferentes das outras pessoas": a vida cotidiana de mulheres com deficiência mental contada por elas mesmas

O objetivo desta pesquisa foi investigar a vida cotidiana de mulheres rotuladas como deficientes mentais, através de seus relatos biográficos pessoais. As entrevistadas descreveram sua rotina em casa e na instituição, falaram sobre suas famílias, relacionamentos, dificuldades em integração social, e sobre seus problemas físicos e de aprendizagem. Ficou clara no discurso deste grupo a dicotomia entre o mundo “de dentro", compreendendo os espaços protegidos da casa e da instituição, e o mundo ameaçador e violento “de fora", representado pela rua. Seus relacionamentos sociais restringem-se aos personagens do mundo “de dentro": a família, os profissionais, e os colegas da instituição, e muitas entrevistadas disseram se· sentir discriminadas pelas pessoas “de fora". Embora várias mulheres tenham exprimido o desejo de ser independentes (trabalhar fora, sair sozinhas, etc), na prática mantém urna relação de extrema dependência familiar. A pesar de dois terços das entrevistadas terem mais de 20 anos, elas não parecem ter nenhuma perspectiva concreta de morar sozinhas, casar, ou vir a formar sua própria família. Um dos pressupostos deste estudo era de que o estigma da deficiência mental, seria o terna central nas histórias de vida. Entretanto, mais da metade dos sujeitos não abordou esta questão, e muito poucas se autodenominaram deficientes. Foi postulado que os efeitos do estigma talvez sejam minimizados neste grupo devido à superproteção familiar e institucional por um lado, e a evitação do mundo "de fora" (onde esta condição seria denunciada mais abertamente) por outro. Assim sendo, em sua prática diária, a maioria dessas mulheres têm poucas oportunidades de se confrontar com a situação de marginalização. Deficiência mental foi analisada como um fenômeno socialmente construído, e acredita-se que estas pessoas funcionem em um nível mais dependente do que sua condição orgânica exigiria, por terem sido reforçadas por representar o papel social de deficientes. Apesar das características comuns, cada história de vida provou ser original e única, mostrando o erro em se considerar as pessoas portadoras de deficiência mental como um grupo homogêneo e bem definido. Além disso, a não ser pela dependência familiar, falta de participação autônoma e integrada na comunidade, e relacionamento amoroso e sexual restrito, a ideia dessas mulheres não é qualitativamente diferente do resto da população.