A novel missense mutation p.L76P in the GJB2 gene causing nonsyndromic recessive deafness in a Brazilian family

Mutations in the GJB2 gene, encoding connexin 26 (Cx26), are a major cause of nonsyndromic recessive hearing loss in many countries. We report here on a novel point mutation in GJB2, p.L76P (c.227C>T), in compound heterozygosity with a c.35delG mutation, in two Brazilian sibs, one presenting mild and the other profound nonsyndromic neurosensorial hearing impairment. Their father, who carried a wild-type allele and a p.L76P mutation, had normal hearing. The mutation leads to the substitution of leucine (L) by proline (P) at residue 76, an evolutionarily conserved position in Cx26 as well as in other connexins. This mutation is predicted to affect the first extracellular domain (EC1) or the second transmembrane domain (TM2). EC1 is important for connexon-connexon interaction and for the control of channel voltage gating. The segregation of the c.227C>T (p.L76P) mutation together with c.35delG in this family indicates a recessive mode of inheritance. The association between the p.L76P mutation and hearing impairment is further supported by its absence in a normal hearing control group of 100 individuals, 50 European-Brazilians and 50 African-Brazilians.

Litter size, effects of maternal body size, and date of birth in South American scorpions (Arachnida: Scorpiones)

We present new data on litter size and date of birth (month) for 21 South American scorpions species. We provide data for one katoikogenic species, the liochelid Opisthacanthus cayaporum Vellard, 1932 (offspring = 3; birth month: Jan); and for several apoikogenic species, such as the bothriurids Bothriurus araguayae Vellard, 1934 (53; Sep), B. rochensis San Martín, 1965 (22-28; Jan, Aug); the buthids Ananteris balzanii Thorell, 1891 (10-34; Jan-Mar), Physoctonus debilis (Koch, 1840) (2; Sep), Rhopalurus amazonicus Lourenço, 1986 (19; Nov), R. lacrau Lourenço & Pinto-da-Rocha, 1997 (30; Dec), R. laticauda Thorell, 1876 (41; Nov), R. rochai Borelli, 1910 (11-47; Dec-Jan, Mar-Apr), Tityus bahiensis (Perty, 1833) (4-23; Oct-Mar), T. clathratus Koch, 1844 (8-18; Nov-Jan), T. costatus (Karsch, 1879) (21-25; Jan, Apr), T. kuryi Lourenço, 1997 (4-16; Mar), T. mattogrossensis Borelli, 1901(8-9; May), T. obscurus (Gervais, 1843) (16-31; Jan-Feb, May, Jul), T. serrulatus Lutz & Mello, 1922 (8-36; Dec, Feb-Apr), T. silvestris Pocock, 1897 (5-14; Dec-Jan, Apr), T. stigmurus (Thorell, 1876) (10-18; Nov, Jan, Mar), Tityus sp. 1 (T. clathratus group - 7-12; Feb-Apr), Tityus sp. 2 (T. bahiensis group - 2; Mar); and the chactid Brotheas sp. (8-21; Jan, Apr). We observed multiple broods: R. lacrau (offspring in the 2nd brood = 27), T. kuryi (6-16), T. obscurus (2-32), T. silvestris (8), T. stigmurus (4-9), T. bahiensis (offspring in the 2nd brood = 2-18; 3rd = 1), and T. costatus (2nd brood = 18; 3rd = 4). We found statistically significant positive correlation between female size and litter size for T. bahiensis and T. silvestris, and nonsignificant correlation for T. serrulatus.

Chemical composition, acetylcholinesterase inhibitory and antifungal activities of Pera glabrata (Schott) Baill. (Euphorbiaceae)

Pera glabrata (Schott) Baill. was selected for this study after showing a preliminary positive result in a screening of Atlantic Forest plant species in the search for acetylcholinesterase inhibitors and antifungal compounds. The bioassays were conducted with crude ethanol extract of the leaves using direct bioautography method for acetylcholinesterase and antifungal activities. This extract was partitioned with hexane, chloroform and ethyl acetate solvents. The active chloroform fraction was submitted to silica gel chromatography column affording 12 groups. Caffeine, an alkaloid, which showed detection limits of 0.1 and 1.0 µg for anticholinesterasic and antifungal activities, respectively, was isolated from group nine. After microplate analyses, only groups four, nine, 10, 11 and 12 showed acetylcholinesterase inhibitory activity of 40% or higher. The group 12 was purified by preparative layer chromatography affording four sub-fractions. Two sub-fractions from this group were analyzed by gas chromatography-mass spectrometry and gas chromatography-flame ionization detector. The first sub-fraction showed anticholinesterasic activity and contained two major compounds: 9-hydroxy-4-megastigmen-3-one (84%) and caffeine (6%). The second sub-fraction presented five major compounds identified as 9-hydroxy-4-megastigmen-3-one, isololiolide, (-) loliolide, palmitic acid and lupeol and did not show activity.

5-HT1A autoreceptor modulation of locomotor activity induced by nitric oxide in the rat dorsal raphe nucleus

The dorsal raphe nucleus (DRN) is the origin of ascending serotonergic projections and is considered to be an important component of the brain circuit that mediates anxiety- and depression-related behaviors. A large fraction of DRN serotonin-positive neurons contain nitric oxide (NO). Disruption of NO-mediated neurotransmission in the DRN by NO synthase inhibitors produces anxiolytic- and antidepressant-like effects in rats and also induces nonspecific interference with locomotor activity. We investigated the involvement of the 5-HT1A autoreceptor in the locomotor effects induced by NO in the DRN of male Wistar rats (280-310 g, N = 9-10 per group). The NO donor 3-morpholinosylnomine hydrochloride (SIN-1, 150, and 300 nmol) and the NO scavenger S-3-carboxy-4-hydroxyphenylglycine (carboxy-PTIO, 0.1-3.0 nmol) were injected into the DRN of rats immediately before they were exposed to the open field for 10 min. To evaluate the involvement of the 5-HT1A receptor and the N-methyl-D-aspartate (NMDA) glutamate receptor in the locomotor effects of NO, animals were pretreated with the 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT, 8 nmol), the 5-HT1A receptor antagonist N-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-N-2-pyridinyl-cyclohexanecarboxamide maleate (WAY-100635, 0.37 nmol), and the NMDA receptor antagonist DL-2-amino-7-phosphonoheptanoic acid (AP7, 1 nmol), followed by microinjection of SIN-1 into the DRN. SIN-1 increased the distance traveled (mean ± SEM) in the open-field test (4431 ± 306.1 cm; F7,63 = 2.44, P = 0.028) and this effect was blocked by previous 8-OH-DPAT (2885 ± 490.4 cm) or AP7 (3335 ± 283.5 cm) administration (P < 0.05, Duncan test). These results indicate that 5-HT1A receptor activation and/or facilitation of glutamate neurotransmission can modulate the locomotor effects induced by NO in the DRN.

Avaliação histológica da neurorrafia término-lateral: estudo experimental em ratos

OBJETIVO: Realizar estudo histológico comparando o crescimento axonal após neurorrafia término-lateral com e sem epineurectomia. MÉTODOS: foram utilizados vinte ratos Wistar, machos, divididos em dois grupos de 10 ratos cada. Um segmento de 1,0cm do nervo tibial e, foi transposto para o lado contralateral, sendo suturado no nervo ciático D. No grupo I, a sutura foi realizada diretamente no epineuro, enquanto que no grupo II foi realizado epineurectomia. Após 4 semanas foi realizado avaliação histológica do segmento transposto e no nervo ciático D, no sitio distal à lesão. RESULTADOS: demonstrou-se baixa quantidade de fibras remielinizadas, variando de 7 a 51 fibras no Grupo I e de 10 a 91 fibras no Grupo II. Utilizou-se o teste U de Mann-Whitney, com p=0,31, demonstrando que não há diferença estatisticamente significante entre os dois grupos. Não há relação positiva entre o número de fibras remielinizadas no enxerto e no sitio distal à lesão do ciático. CONCLUSÃO: A neurorrafia término-lateral, com e sem janela epineural, não promove remielinização eficiente. Nivel de evidência: Nível II: Estudo prospectivo comparativo

Análise dos efeitos da cinesioterapia e da hidrocinesioterapia sobre a qualidade de vida de pacientes com fibromialgia: um ensaio clínico randomizado

OBJETIVO: Verificar e comparar os efeitos da hidrocinesioterapia e da cinesioterapia na qualidade de vida de pacientes portadoras de fibromialgia. MATERIAIS E MÉTODOS: Foi realizado um ensaio clínico randomizado, cego, para os avaliadores. Para avaliar a qualidade de vida, utilizou-se o questionário genérico SF-36 a fim de analisar os aspectos físicos e psicológicos em 24 pacientes do sexo feminino, portadoras de fibromialgia, com idade de 30 a 55 anos, alocadas aleatoriamente em dois grupos: hidrocinesioterapia e cinesioterapia. Durante um período de 23 semanas, as pacientes foram submetidas a alongamentos e exercícios aeróbios de baixa intensidade com duração de uma hora cada sessão (em ambos os grupos). RESULTADOS: Observou-se melhora estatisticamente significante (p < 0,05) na maior parte dos aspectos abordados pelo SF-36 em ambos os grupos. CONCLUSÃO: A hidrocinesioterapia promoveu um relaxamento muscular favorável para a melhora da qualidade de vida, porém, não foi o fator principal para amenizar os efeitos da patologia, uma vez que a cinesioterapia também proporcionou efeitos benéficos para a promoção do bem-estar dessas pacientes. Portanto, os alongamentos e os exercícios aeróbios de baixa intensidade empregados em ambos os protocolos são os prováveis responsáveis pelos efeitos benéficos observados nas duas modalidades terapêuticas analisadas.

Induction of Zenk protein expression within the nucleus taeniae of the amygdala of pigeons following tone and shock stimulation

In this study, we evaluated the expression of the Zenk protein within the nucleus taeniae of the pigeon’s amygdala (TnA) after training in a classical aversive conditioning, in order to improve our understanding of its functional role in birds. Thirty-two 18-month-old adult male pigeons (Columba livia), weighing on average 350 g, were trained under different conditions: with tone-shock associations (experimental group; EG); with shock-alone presentations (shock group; SG); with tone-alone presentations (tone group; TG); with exposure to the training chamber without stimulation (context group; CG), and with daily handling (naive group; NG). The number of immunoreactive nuclei was counted in the whole TnA region and is reported as density of Zenk-positive nuclei. This density of Zenk-positive cells in the TnA was significantly greater for the EG, SG and TG than for the CG and NG (P < 0.05). The data indicate an expression of Zenk in the TnA that was driven by experience, supporting the role of this brain area as a critical element for neural processing of aversive stimuli as well as meaningful novel stimuli.

Essential regulatory elements for NHE3 gene transcription in renal proximal tubule cells

The objectives of the present study were to identify the cis-elements of the promoter absolutely required for the efficient rat NHE3 gene transcription and to locate positive and negative regulatory elements in the 5’-flanking sequence (5’FS), which might modulate the gene expression in proximal tubules, and to compare this result to those reported for intestinal cell lines. We analyzed the promoter activity of different 5’FS segments of the rat NHE3 gene, in the OKP renal proximal tubule cell line by measuring the activity of the reporter gene luciferase. Because the segment spanning the first 157 bp of 5’FS was the most active it was studied in more detail by sequential deletions, point mutations, and gel shift assays. The essential elements for gene transcription are in the region -85 to -33, where we can identify consensual binding sites for Sp1 and EGR-1, which are relevant to NHE3 gene basal transcription. Although a low level of transcription is still possible when the first 25 bp of the 5’FS are used as promoter, efficient transcription only occurs with 44 bp of 5’FS. There are negative regulatory elements in the segments spanning -1196 to -889 and -467 to -152, and positive enhancers between -889 and -479 bp of 5’FS. Transcription factors in the OKP cell nuclear extract efficiently bound to DNA elements of rat NHE3 promoter as demonstrated by gel shift assays, suggesting a high level of similarity between transcription factors of both species, including Sp1 and EGR-1.

Physiological implications of the regulation of vacuolar H+-ATPase by chloride ions

Vacuolar H+-ATPase is a large multi-subunit protein that mediates ATP-driven vectorial H+ transport across the membranes. It is widely distributed and present in virtually all eukaryotic cells in intracellular membranes or in the plasma membrane of specialized cells. In subcellular organelles, ATPase is responsible for the acidification of the vesicular interior, which requires an intraorganellar acidic pH to maintain optimal enzyme activity. Control of vacuolar H+-ATPase depends on the potential difference across the membrane in which the proton ATPase is inserted. Since the transport performed by H+-ATPase is electrogenic, translocation of H+-ions across the membranes by the pump creates a lumen-positive voltage in the absence of a neutralizing current, generating an electrochemical potential gradient that limits the activity of H+-ATPase. In many intracellular organelles and cell plasma membranes, this potential difference established by the ATPase gradient is normally dissipated by a parallel and passive Cl- movement, which provides an electric shunt compensating for the positive charge transferred by the pump. The underlying mechanisms for the differences in the requirement for chloride by different tissues have not yet been adequately identified, and there is still some controversy as to the molecular identity of the associated Cl--conducting proteins. Several candidates have been identified: the ClC family members, which may or may not mediate nCl-/H+ exchange, and the cystic fibrosis transmembrane conductance regulator. In this review, we discuss some tissues where the association between H+-ATPase and chloride channels has been demonstrated and plays a relevant physiologic role.

Evidence for divided automatic attention

A long-standing debate in the literature is whether attention can form two or more independent spatial foci in addition to the well-known unique spatial focus. There is evidence that voluntary visual attention divides in space. The possibility that this also occurs for automatic visual attention was investigated here. Thirty-six female volunteers were tested. In each trial, a prime stimulus was presented in the left or right visual hemifield. This stimulus was characterized by the blinking of a superior, middle or inferior ring, the blinking of all these rings, or the blinking of the superior and inferior rings. A target stimulus to which the volunteer should respond with the same side hand or a target stimulus to which she should not respond was presented 100 ms later in a primed location, a location between two primed locations or a location in the contralateral hemifield. Reaction time to the positive target stimulus in a primed location was consistently shorter than reaction time in the horizontally corresponding contralateral location. This attentional effect was significantly smaller or absent when the positive target stimulus appeared in the middle location after the double prime stimulus. These results suggest that automatic visual attention can focus on two separate locations simultaneously, to some extent sparing the region in between.

Estudo da fauna de triatomíneos e da ocorrência de doença de Chagas em Monte Negro, Rondônia, Brasil

A doença de Chagas tem como agente etiológico o Trypanossoma cruzi, um protozoário flagelado que pode ser encontrado numa grande variedade de mamíferos e triatomíneos. O Estado de Rondônia, localizado na Amazônia Ocidental, possui um meio ambiente constantemente modificado pelas ações transformadoras do ser humano, resultando em um desequilíbrio, que pode facilitar a transmissão de inúmeros patógenos. Uma grande variedade e quantidade de palmáceas, em especial o babaçu, bem como mamíferos e triatomíneos, podem ser encontrados neste complexo ecossistema. Nesta pesquisa, a fauna de triatomíneos foi identificada em 225 babaçus e por meio de capturas peri e intradomiciliares. Foi realizado, concomitantemente, estudo de soroprevalência para doença de Chagas e a identificação da presença de T.cruzi no trato digestivo dos triatomíneos. Positividade ao T. cruzi foi verificada em 23,7% dos 652 triatomíneos coletados nos babaçus. Estes triatomíneos pertenciam ao gênero Rhodnius e foram classificados em 4 espécies: R. robustus, R. prolixus, R. pictipes e R. milesi. Nas capturas intradomiciliares, dez espécimes do Rhodnius robustus e uma de Panstrongylus geniculattus foram encontrados, sendo que 3% da população foi positiva para doença de Chagas. Na área pesquisada, há potencial de transmissão da doença de Chagas na forma endêmica devido a grande quantidade de triatomíneos, bem como alta freqüência de infecção destes triatomíneos, porém no momento deste estudo não se evidenciou a ocorrência da transmissão.

Antibody responses elicited in mice immunized with Bacillus subtilis vaccine strains expressing Stx2B subunit of enterohaemorragic Escherichia coli O157:H7

No effective vaccine or immunotherapy is presently available for patients with the hemolytic uremic syndrome (HUS) induced by Shiga-like toxin (Stx) producedbyenterohaemorragic Escherichia coli (EHEC) strains, such as those belonging to the O157:H7 serotype. In this work we evaluated the performance of Bacillus subtilis strains, a harmless spore former gram-positive bacterium species, as a vaccine vehicle for the expression of Stx2B subunit (Stx2B). A recombinant B. subtilis vaccine strain expressing Stx2B under the control of a stress inducible promoter was delivered to BALB/c mice via oral, nasal or subcutaneous routes using both vegetative cells and spores. Mice immunized with vegetative cells by the oral route developed low but specific anti-Stx2B serum IgG and fecal IgA responses while mice immunized with recombinant spores developed anti-Stx2B responses only after administration via the parenteral route. Nonetheless, serum anti-Stx2B antibodies raised in mice immunized with the recombinant B. subtilis strain did not inhibit the toxic effects of the native toxin, both under in vitro and in vivo conditions, suggesting that either the quantity or the quality of the induced immune response did not support an effective neutralization of Stx2 produced by EHEC strains.

Rates of antimicrobial resistance in latin america (2004-2007) and in vitro activity of the glycylcycline tigecycline and of other antibiotics

As a part of the Tigecycline Evaluation and Surveillance Trial (T.E.S.T.), Gram-positive and Gram-negative bacterial isolates were collected from 33 centers in Latin America (centers in Argentina, Brazil, Chile, Colombia, Guatemala, Honduras, Jamaica, Mexico, Panama, Puerto Rico, and Venezuela) from January 2004 to September 2007. Argentina and Mexico were the greatest contributors of isolates to this study. Susceptibilities were determined according to Clinical Laboratory Standards Institute guidelines. Resistance levels were high for most key organisms across Latin America: 48.3% of Staphylococcus aureus isolates were methicillin-resistant while 21.4% of Acinetobacter spp. isolates were imipenem-resistant. Extended-spectrum β-lactamase were reported in 36.7% of Klebsiella pneumoniae and 20.8% of E. coli isolates. Tigecycline was the most active agent against Gram-positive isolates. Tigecycline was also highly active against all Gram-negative organisms, with the exception of Pseuodomonas aeruginosa, against which piperacillin-tazobactam was the most active agent tested (79.3% of isolates susceptible). The in vitro activity of tigecycline against both Gram-positive and Gram-negative isolates indicates that it may be an useful tool for the treatment of nosocomial infections, even those caused by organisms that are resistant to other antibacterial agents.

Cation size effects-modified phase and PTCR development in Er3+ and Ca2+ co-doped BaTiO3 ceramics during sintering

Development of the positive temperature coefficient of resistivity (PTCR) in Er3+ and Ca2+ co-doped ferroelectric BaTiO3 was studied in this work, with Er3+ being used to act as a donor doping. Irrespective of all the materials showing high densities after sintering at 1200 to 1300 ºC, these revealed insulator at the lowest sintering temperature, changing to semiconducting and PTCR-type materials only when the sintering temperature was further increased. Observations from X-ray diffraction help correlating this effect with phase development in this formulated (Ba,Ca,Er)TiO3 system, considering the formation of initially two separated major (Ba,Ca)TiO3- and minor (Ca,Er)TiO3-based compounds, as a consequence of cation size-induced stress energy effects. Thus, appearance and enhancement here of the semiconducting and PTCR responses towards higher sintering temperatures particularly involve the incorporation of Er3+ into the major phase, rendering finally possible the generation and "percolative-like" migration of electrons throughout the whole material.

Multimodos e múltiplas Representações, aprendizagem significativa e subjetividade: três referenciais conciliáveis da educação científica

A linha de pesquisa em multimodos e múltiplas representações vem atualmente sendo inspiradora de ações instrucionais na educação científica. Partindo dos fundamentos que justificam um encaminhamento didático à luz dessas referências, este trabalho procura mostrar que há compatibilidade dos seus fundamentos com a teoria da aprendizagem significativa de Ausubel e com as questões levantadas pelas pesquisas que indicam a necessidade de se considerar a subjetividade dos alunos presentes numa sala de aula. Essencialmente, procuramos argumentar que a promoção de um ensino por meio de multimodos e múltiplas representações é consistente com o ambiente plural das subjetividades existentes numa sala de aula e com uma aprendizagem significativa.