Novel Mutations in CYP11B1 Gene Leading to 11 beta-Hydroxylase Deficiency in Brazilian Patients

Background: Deficiency of 11 beta-hydroxylase results in the impairment of the last step of cortisol synthesis. In females, the phenotype of this disorder includes different degrees of genital ambiguity and arterial hypertension. Mutations in the CYP11B1 gene are responsible for this disease. Objective: The objective of the study was to screen the CYP11B1 gene for mutations in two unrelated Brazilian females with congenital adrenal hyperplasia due to 11 beta-hydroxylase deficiency. Design: The coding and intron-exon junction regions of CYP11B1 were totally sequenced. A putative splice mutation was further investigated by minigene transcription. Results: We report two novel CYP11B1 mutations in these Brazilian patients. An Arabian Lebanese descendent female was found to be homozygous for a cytosine insertion at the beginning of exon 8, changing the 404 arginine to proline. It alters the open reading frame, creating a putative truncated protein at 421 residue, which eliminates the domain necessary for the association of heme prosthetic group. A severely virilized female was homozygous for the g. 2791G>A transition in the last position of exon 4. This nucleotide is also part of 5` intron 4 donor splice site consensus sequence. Minigene experiments demonstrated that g. 2791G>A activated an alternative splice site within exon 4, leading to a 45-bp deletion in the transcript. The putative translation of such modified mRNA indicates a truncated protein at residue 280. Conclusions: We describe two novel mutations, g. 4671_4672insC and g. 2791G>A, that drastically affects normal protein structure. These mutations abolish normal enzyme activity, leading to a severe phenotype of congenital adrenal hyperplasia due to 11 beta-hydroxylase deficiency. (J Clin Endocrinol Metab 94: 3481-3485, 2009)

The Effect of Repetitive Pilot-Hole Use on the Insertion Torque and Pullout Strength of Vertebral System Screws

Study Design. In vitro biomechanical investigation of the screw-holding capacity. Objective. To evaluate the effect of repetitive screw-hole use on the insertional torque and retentive strength of vertebral system screws. Summary and Background Data. Placement and removal of vertebral system screws is sometimes necessary during the surgical procedures in order to assess the walls of the pilot hole. This procedure may compromise the holding capacity of the implant. Methods. Screws with outer diameter measuring 5, 6, and 7 mm were inserted into wood, polyurethane, polyethylene, and cancellous bone cylindrical blocks. The pilot holes were made with drills of a smaller, equal, or wider diameter than the inner screw diameter. Three experimental groups were established based on the number of insertions and reinsertions of the screws and subgroups were created according to the outer diameter of the screw and the diameter of the pilot hole used. Results. A reduction of screw-holding capacity was observed between the first and the following insertions regardless the anchorage material. The pattern of reduction of retentive strength was not similar to the pattern of torque reduction. The pullout strength was more pronounced between the first and the last insertions, while the torque decreased more proportionally from the first to the last insertions. Conclusion. Insertion and reinsertion of the screws of the vertebral fixation system used in the present study reduced the insertion torque and screw purchase.

Day-night pattern of autonomic nervous system modulation in patients with heart failure with and without sleep apnea

Introduction: Among patients with congestive heart failure (CHF) both obstructive and central sleep apnea (SA) are associated with increased sympathetic activity. However, the day-night pattern of cardiac autonomic nervous system modulation in CHF patients with and without sleep apnea is unknown. Material and methods: Twenty-five CHF patients underwent polysomnography with simultaneous beat-to-beat blood pressure (Portapres), respiration and electrocardiogram monitoring. Patients were divided according to the presence (SA, n=17) and absence of SA (NoSA, n=8). Power spectral analyses of heart rate variability (HRV) and spontaneous baroreflex sensitivity (BRS) were determined in periods with stable breathing while awake at 6 AM, 10 AM, 10 PM, as well as during stage 2 sleep. In addition, muscle sympathetic nerve activity (MSNA) was evaluated at 10 AM. Results: RR variance, low-frequency (LF), high-frequency (HF) powers of HRV, and BRS were significantly lower in patients with SA compared with NoSA in all periods. HF power, a marker of vagal activity, increased during sleep in patients with NoSA but in contrast did not change across the 24-hour period in patients with SA. MSNA was significantly higher in patients with SA compared with NoSA. RR variance, LF and HF powers correlated inversely with simultaneous MSNA (r=-0.64, -0.61, and -0.61 respectively; P < 0.01). Conclusions: Patients with CHF and SA present a reduced and blunted cardiac autonomic modulation across the 24-hour period. These findings may help to explain the increased cardiovascular risk in patients with CHF and SA. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Comparison of the effects of aerobic and resistance training on cardiac autonomic adaptations in ovariectomized rats

We have compared the effects of two types of physical training on the cardiac autonomic control in ovariectomized and sham-operated rats according to different approaches: double autonomic blockade (DAB) with methylatropine and propranolol; baroreflex sensibility (BRS) and spectral analysis of heart rate variability (HRV). Wistar female rats (+/- 250 g) were divided into two groups: sham-operated and ovariectomized. Each group was subdivided into three subgroups: sedentary rats, rats submitted to aerobic trained and rats submitted to resistance training. Ovariectomy did not change arterial pressure, basal heart rate (HR), DAB and BRS responses, but interfered with HRV by reducing the low-frequency oscillations (LF = 0.20-0.75 Hz) in relation to sedentary sham-operated rats. The DAB showed that both types of training promoted an increase in the predominance of vagal tonus in sham-operated rats, but HR variations due to methylatropine were decreased in the resistance trained rats compared to sedentary rats. Evaluation of BRS showed that resistance training for sham-operated and ovariectomized rats reduced the tachycardic responses in relation to aerobic training. Evaluation of HRV in trained rats showed that aerobic training reduced LF oscillations in sham-operated rats, whereas resistance training had a contrary effect. In the ovariectomized rats, aerobic training increased high frequency oscillations (HF = 0.75-2.5 Hz), whereas resistance training produced no effect. In sham-operated rats, both types of training increased the vagal autonomic tonus, but resistance training reduced HF oscillations and BRS as well. In turn, both types of training had similar results in ovariectomized rats, except for HRV, as aerobic training promoted an increase in HF oscillations. (C) 2011 Elsevier B.V. All rights reserved.

The effect of ovariectomy on cardiac autonomic control in rats submitted to aerobic physical training

We have investigated the ovariectomy effects on the cardiovascular autonomic adaptations induced by aerobic physical training and the role played by nitric oxide (NO). Female Wistar rats (n =70) were divided into five groups: Sedentary Sham (SS): Trained Sham (TS); Trained Hypertensive Sham treated with N(C)-nitro-L-arginine methyl ester (L-NAME) (THS): Trained Ovariectomized (TO); and Trained Hypertensive Ovariectomized treated with L-NAME (THO). Trained groups were submitted to a physical training during 10 weeks. The cardiovascular autonomic control was investigated in all groups using different approaches: 1) pharmacological evaluation of autonomic tonus with methylatropine and propranolol; 2) analysis of heart rate (HR) and systolic arterial pressure (AP) variability; 3) spontaneous baroreflex sensitivity (BRS) evaluation. Hypertension was observed in THS and THO groups. Pharmacological analysis showed that TS group had increased predominance of autonomic vagal tonus compared to SS group. HR and intrinsic HR were found to be reduced in all trained animals. TS group, compared to other groups, showed a reduction in LF oscillations (LF=0.2-0.75 Hz) of pulse interval in both absolute and normalized units as well as an increase in HF oscillations (HF=0.75-2.50 Hz) in normalized unit. FIRS analysis showed that alpha-index was different between all groups. TS group presented the greatest value, followed by the TO, SS. THO and THS groups. Ovariectomy has negative effects on cardiac autonomic modulation in trained rats, which is characterized by an increase in the sympathetic autonomic modulation. These negative effects suggest NO deficiency. In contrast, the ovariectomy seems to have no effect on AP variability. (C) 2008 Elsevier B.V. All rights reserved.

Effect of 4% titanium tetrafluoride solution on dental erosion by a soft drink: An in situ/ex vivo study

Objective: This in situ/ex vivo study assessed the effect of titanium tetrafluoride (TiF4) on permanent human enamel subjected to erosion. Design: Ten volunteers took part in this study performed in two phases. In the first phase (ERO), they wore acrylic palatal appliances containing two enamel blocks, divided into two rows: TiF4 (F) and no-TiF4 (no-F). During the 1st day, the formation of a salivary pellicle was allowed. In the 2nd day, the TiF4 solution was applied on one row (ERO + F), whereas on the other row no treatment was performed (ERO + no-F). From 3rd until 7th day, the blocks were subjected to erosion, 4x per day. In the 2nd phase (no-ERO), the volunteers wore acrylic palatal appliances containing one enamel block, during 2 days, to assess the effect of TiF4 only (no-ERO + F). Enamel alterations were determined using profilometry (wear), microhardness (%SMHC) tests, scanning electron microscope and microprobe analysis. The %SMHC and wear were tested using ANOVA and Tukey`s post hoc tests (p < 0.05). Results: The mean of %SMHC and wear ( mu m) values ( +/- S.D.) were, respectively: ERO + F -73.32 +/- 5.16(A)/2.40 +/- 0.60(a); ERO + no-F -83.49 +/- 4.59B/1.17 +/- 0.48(b) and no-ERO + F -67.92 +/- 6.16(A)/0.21:E 0.09(c). In microscope analysis, the no-F group showed enamel with honeycomb appearance. For F groups, it was observed a surface coating with microcracks. The microprobe analysis revealed the presence of the following elements (%) in groups ERO + F, ERO + no-F and no-ERO + F, respectively: Ca (69.9, 72.5, 66.25); P (25.9, 26.5, 26.06); Ti (3.0, 0, 5.93). Conclusions: The TiF4 was unable to reduce dental erosion. (c) 2007 Elsevier Ltd. All rights reserved.

In Vitro Proliferation and Osteoblastic Phenotype Expression of Cells Derived From Human Vertebral Lamina and Iliac Crest

Study Design. Osteoblastic cells derived from vertebral lamina and iliac crest were isolated and cultured under the same conditions (osteogenic medium, pH, temperature, and CO(2) levels). Objective. To compare proliferation and expression of osteoblastic phenotype of cells derived from vertebral lamina and iliac grafting. Summary of Background Data. Many factors play a role in the success of bone graft in spinal fusion including osteoblastic cell population. Two common sources of graft are vertebral lamina and iliac crest, however, differences in proliferation and osteoblastic phenotype expression between cells from these sites have not been investigated. Methods. Cells obtained from cancellous bone of both vertebral lamina and iliac crest were cultured and proliferation was evaluated by direct cell counting and viability detected by Trypan blue. Alkaline phosphatase (ALP) activity was evaluated by thymolphthalein release from thymolphthalein monophosphate and matrix mineralization by staining with alizarin red S. Gene expression of ALP, osteocalcin, runt-related transcription factor 2, Msh homeobox 2, bone morphogenetic protein 7, intercellular adhesion molecule 1 precursor, osteoprotegerin, and receptor activator of NF-kB ligand was analyzed by real-time PCR. All comparisons were donor-matched. Results. Proliferation was greater at days 7 and 10 in cells from vertebral lamina compared with ones from iliac crest without difference in cell viability. ALP activity was higher in cells from vertebral lamina compared with cells from iliac crest at days 7 and 10. At 21 days, mineralized matrix was higher in cells derived from vertebral lamina than from iliac crest. At day 7, gene expression of ALP, osteocalcin, runt-related transcription factor 2, Msh homeobox 2, bone morphogenetic protein 7, intercellular adhesion molecule 1 precursor, receptor activator of NF-kB ligand, and osteoprotegerin was higher in cells derived from vertebral lamina compared with iliac crest. Conclusion. Cell proliferation and osteoblastic phenotype development in cells derived from cancellous bone were more exuberant in cultures of vertebral lamina than of iliac crest.

'Neighbourhood' size, dispersal and density estimates in the prickly forest skink (Gnypetoscincus queenslandiae) using individual genetic and demographic methods

Dispersal, or the amount of dispersion between an individual's birthplace and that of its offspring, is of great importance in population biology, behavioural ecology and conservation, however, obtaining direct estimates from field data on natural populations can be problematic. The prickly forest skink, Gnypetoscincus queenslandiae, is a rainforest endemic skink from the wet tropics of Australia. Because of its log-dwelling habits and lack of definite nesting sites, a demographic estimate of dispersal distance is difficult to obtain. Neighbourhood size, defined as 4 piD sigma (2) (where D is the population density and sigma (2) the mean axial squared parent-offspring dispersal rate), dispersal and density were estimated directly and indirectly for this species using mark-recapture and microsatellite data, respectively, on lizards captured at a local geographical scale of 3 ha. Mark-recapture data gave a dispersal rate of 843 m(2)/generation (assuming a generation time of 6.5 years), a time-scaled density of 13 635 individuals * generation/km(2) and, hence, a neighbourhood size of 144 individuals. A genetic method based on the multilocus (10 loci) microsatellite genotypes of individuals and their geographical location indicated that there is a significant isolation by distance pattern, and gave a neighbourhood size of 69 individuals, with a 95% confidence interval between 48 and 184. This translates into a dispersal rate of 404 m(2)/generation when using the mark-recapture density estimation, or an estimate of time-scaled population density of 6520 individuals * generation/km(2) when using the mark-recapture dispersal rate estimate. The relationship between the two categories of neighbourhood size, dispersal and density estimates and reasons for any disparities are discussed.

An??lise da estrutura organizacional da fiscaliza????o do transporte rodovi??rio na ag??ncia nacional de transportes terrestres

Fiscalizar a presta????o dos servi??os de transporte interestadual e internacional de passageiros e de transporte rodovi??rio de cargas ?? atribui????o da Superintend??ncia de Fiscaliza????o da Ag??ncia Nacional de Transportes Terrestres ??? ANTT. Esta pesquisa visa analisar a efici??ncia e efic??cia desta estrutura organizacional

Modernizando a administra????o p??blica na Am??rica Latina: problemas comuns sem solu????es f??ceis

A inten????o dos autores ?? fornecer subs??dios para o debate sobre a reforma da administra????o p??blica na Am??rica Latina. O artigo come??a pela caracteriza????o dos desafios colocados para a burocracia pelos problemas do monop??lio e do controle pol??tico. Em seguida, s??o analisados os modelos de administra????o p??blica adotados pelos pa??ses desenvolvidos, dado que estes constituem os principais pontos de refer??ncia para as iniciativas de reforma nos pa??ses em desenvolvimento. E, finalmente, os autores identificam as causas e os efeitos das disfun????es da administra????o p??blica na Am??rica Latina, sugerindo que as experi??ncias e o debate acad??mico relativos aos pa??ses desenvolvidos podem fornecer li????es aos pa??ses em desenvolvimento desde que n??o se perca de vista as especificidades do contexto latino-americano. Os autores sustentam que nestes pa??ses as iniciativas de reforma administrativa s??o dificultadas pelo baixo n??vel de desenvolvimento pol??tico e pela vig??ncia de padr??es informais de comportamento no ??mbito da burocracia p??blica.

Cultivares de feijoeiro: efeito do solo, adubação foliar e competição com trapoeraba

O feijoeiro (Phaseolus vulgaris L.) é um dos principais alimentos consumidos no Brasil, rico em vitaminas, carboidratos e minerais. É crescente o número de pesquisas que integram desde o melhoramento de plantas e o manejo da adubação para incrementar nutrientes nas partes comestíveis, desenvolvendo plantas com maiores teores de vitaminas e micronutrientes. A introdução de alimentos biofortificados como o feijão, complementa a nutrição humana que sofre com desnutrição. Sendo assim, é importante conhecer como as condições de cultivo influenciam na qualidade do grão e na importância para obtenção de um alimento com maior valor nutricional. Além de conhecer até que ponto a influência de planta daninha afeta a produção e a absorção de nutrientes pelo feijoeiro. O objetivo deste trabalho foi analisar as características agronômicas e nutricionais influenciadas pela interferência de planta daninha, efeito da adubação e do solo, nas cultivares de feijão. Foi avaliado o teor de clorofila, número de folhas, diâmetro do caule, número de vagem por planta, índice de colheita por planta, massa seca da trapoeraba, relação grão e lócus, peso médio de sementes e análise de ferro e zinco nas folhas e nos grãos. Os resultados obtidos no estudo apontaram que a competição com trapoeraba afetou algumas características agronômicas, devido à competição por nutriente. O solo para cultivo também interferiu na produção, o solo eutrófico proporcionou melhores resultados das cultivares. A adubação não interferiu nos teores de ferro e zinco nos grãos do feijoeiro. As cultivares BRS Agreste, BRS Ametista e BRS Estilo apresentaram melhores resultados na maioria das características analisadas. Por fim, conclui-se que a produção do feijoeiro, necessita de um solo com uma boa nutrição, um bom teor de matéria orgânica, manejo da adubação e de plantas daninhas para aumentar a produção.

Tolerância à salinidade avaliada em genótipos de arroz cultivados in vitro

As plantas em condições naturais estão expostas a vários estresses ambientais que afetam seu metabolismo. Dentre esses, a salinidade dos solos e da água de irrigação é um dos mais sérios problemas para a agricultura irrigada. O objetivo deste trabalho foi identificar, por meio de caracteres morfológicos, a variabilidade genética de 10 genótipos de arroz, cultivados in vitro, e agrupar esses genótipos para o caráter tolerância à salinidade. Os tratamentos foram constituídos por 10 genótipos e quatro concentrações de NaCl (0, 4, 8 e 12 mg L-1) acrescidas ao meio de cultura MS. Após 21 dias, foram avaliados diversos caracteres morfológicos, para os quais foram realizados cálculos percentuais de desempenho relativo (aumento ou redução), considerando-se o valor absoluto do tratamento-controle (0 mg L-1). Todos os caracteres mensurados tiveram seu desenvolvimento reduzido em substrato salino, sendo os correspondentes à biomassa média da parte aérea e do sistema radicular os mais sensíveis ao NaCl. Observou-se dissimilaridade entre os genótipos estudados para tolerância à salinidade, verificada pela formação de três grupos distintos pelo método hierárquico UPGMA e dois grupos pelo método de Tocher, sendo o genótipo BRS Bojuru o mais tolerante e BRS "7" Taim e BRS Ligeirinho os mais sensíveis à salinidade.

Eficiência no uso de fósforo de variedades de arroz cultivadas em solos de várzea irrigada

A cultura do arroz destaca-se como uma das mais importantes do mundo, por apresentar facilidade de adaptação a condições edafoclimáticas distintas. Cultivado e consumido em todos continentes, o arroz destaca-se pela produção e área de cultivo, desempenhando papel estratégico tanto em nível econômico quanto social. No Brasil, a maior parcela da produção de arroz é proveniente do ecossistema de várzea. Este trabalho objetivou estudar a eficiência e resposta ao uso de fósforo de variedades de arroz (Oryza sativa L.), em várzea irrigada, no Sudoeste do Estado de Tocantins. Os tratamentos envolveram oito variedades comerciais de arroz (BRS-Jaçanã, Best-2000, BRS-Guará, BRS-Alvorada, BRA-01381, AN-Cambará, BRS 7-Taim e EPAGRI-109), que foram cultivadas em dois ambientes distintos. Para simular ambientes com baixo e alto níveis de fósforo, foram utilizadas as doses de 20 e 120 kg ha-1 de P2O5, respectivamente. O delineamento experimental utilizado foi o de blocos casualizados, com quatro repetições. Por meio da produtividade de grãos, as variedades foram classificadas quanto à eficiência no uso e resposta à aplicação de fósforo. Demonstrou-se que apenas a variedade BRS-Alvorada é eficiente quanto ao uso de fósforo e responsiva a sua aplicação.

Crescimento de dois cultivares de soja submetidos a aplicações de herbicidas e fungicidas

Existem relatos de que a aplicação de fungicidas do grupo das estrobilurinas na cultura da soja tem trazido aumento de produtividade mesmo em locais sem incidência de doenças, indicando que as alterações de ordem fisiológica devem ser quantificadas. Este estudo foi desenvolvido com o objetivo de avaliar o crescimento de dois cultivares de soja (MG/BR46 Conquista e BRS Valiosa RR) submetidos a aplicações de herbicidas e fungicidas (estrobilurinas e/ou triazóis). Dois experimentos foram desenvolvidos em campo com esquema de tratamentos em parcelas subdivididas no tempo (5 x 12), em que nas parcelas foram alocadas cinco combinações de herbicidas (glyphosate a 720 g ha-1 ou sethoxydim (230 g ha-1) + bentazon + chlorimuron-ethyl (480 + 11,25 g ha-1)) e fungicidas (pyraclostrobin a 75 g ha-1, epoxiconazole a 50 g ha-1 ou pyraclostrobin + epoxiconazole (66,5 + 25 g ha-1)); e como subparcelas foram consideradas as datas de avaliação entre 30 e 120 dias após semeadura. A aplicação de herbicidas não alterou o crescimento dos cultivares de soja. A aplicação de fungicidas do grupo das estrobilurinas associado a triazóis promoveu maior acúmulo de massa de matéria seca e área foliar. Ainda a aplicação desses fungicidas em R2 (pleno florescimento) e R5.1 (enchimento de grãos) resultou em maiores valores de taxas de crescimentos absoluto e relativo, assimilatória líquida e de crescimento da cultura em período próximo ao final do ciclo da cultura, diferenciando, inclusive, dos tratamentos com aplicação somente de triazóis.

Preferência de Aphis craccivora por genótipos de feijão-caupi de porte prostrado, em Aquidauana, MS

O feijão-caupi (Vigna unguiculata (L.) Walp.) é uma importante fonte de alimento. Na região centrooeste do Brasil, poucos são os estudos desenvolvidos com essa cultura. Assim, foram desenvolvidos experimentos, em condições de campo, em Aquidauana, MS, visando encontrar genótipos menos preferidos pelo pulgão preto (Aphis craccivora) para infestação e colonização. Em abril e outubro de 2009, foram semeados 20 genótipos de porte prostrado e após a germinação verificouse o número de fêmeas e colônias de pulgão presentes e aos 30 dias avaliouse a porcentagem de plantas infestadas. No experimento conduzido em abril e maio, verificouse que todos os genótipos de feijãocaupi foram colonizados por A. craccivora. Porém nos genótipos MNC99537F142 e BRS Paraguassu foram observadas as menores percentagens de colônias. Em maio encontrouse as menores percentagens nos genótipos MNC99537F142, MNC01631F205 e BRS Paraguassu, com 6,3, 5,7 e 6,3% de plantas infestadas, respectivamente. Dentre os genótipos estudados, concluise que MNC99537F142, MNC01631F205 e BRS Paraguassu, podem ser melhores estudados, visando à seleção de materiais resistentes ao pulgão preto.