A novel DAX1/NR0B1 mutation in a patient with adrenal hypoplasia congenita and hypogonadotropic hypogonadism

We report a case of adrenal hypoplasia congenita (AHC) and hypogonadotropic hypogonadism (HH) due to a novel DAX1 mutation. A 19-month-old boy with hyperpigmentation and failure to thrive came to our service for investigation. Three brothers of the patient had died due to adrenal failure, and a maternal cousin had adrenal insufficiency. Adrenoleukodystrophy was excluded. MRI showed normal pituitary and hypothalamus. Plasma hormone evaluation revealed high ACTH (up to 2,790 pg/mL), and low levels of androstenedione, DHEA-S, 11-deoxycortisol, and cortisol. At 14 years of age the patient was still prepubescent, his weight was 43.6 kg (SDS: -0.87) and his height was 161 cm (SDS: -0.36), with normal body proportions. In the GnRH test, basal and maximum values of LH and FSH were respectively 0.6/2.1 and < 1.0/< 1.0 U/L. Molecular investigation identified a novel mutation that consists of a deletion of codon 372 (AAC; asparagine) in exon 1 of DAX1. This mutation was not found in a study of 200 alleles from normal individuals. Prediction site analysis indicated that this alteration, located in the DAX1 ligand-binding domain, may damage DAX1 protein. We hypothesize that the novel (p.Asp372del) DAX1 mutation might be able to cause a disruption of DAX1 function, and is probably involved in the development of AHC and HH in this patient. Arq Bras Endocrinol Metab. 2012;56(8):496-500

Comparative EST analysis provides insights into the basal aquatic fungus Blastocladiella emersonii

Abstract Background Blastocladiella emersonii is an aquatic fungus of the Chytridiomycete class, which is at the base of the fungal phylogenetic tree. In this sense, some ancestral characteristics of fungi and animals or fungi and plants could have been retained in this aquatic fungus and lost in members of late-diverging fungal species. To identify in B. emersonii sequences associated with these ancestral characteristics two approaches were followed: (1) a large-scale comparative analysis between putative unigene sequences (uniseqs) from B. emersonii and three databases constructed ad hoc with fungal proteins, animal proteins and plant unigenes deposited in Genbank, and (2) a pairwise comparison between B. emersonii full-length cDNA sequences and their putative orthologues in the ascomycete Neurospora crassa and the basidiomycete Ustilago maydis. Results Comparative analyses of B. emersonii uniseqs with fungi, animal and plant databases through the two approaches mentioned above produced 166 B. emersonii sequences, which were identified as putatively absent from other fungi or not previously described. Through these approaches we found: (1) possible orthologues of genes previously identified as specific to animals and/or plants, and (2) genes conserved in fungi, but with a large difference in divergence rate in B. emersonii. Among these sequences, we observed cDNAs encoding enzymes from coenzyme B12-dependent propionyl-CoA pathway, a metabolic route not previously described in fungi, and validated their expression in Northern blots. Conclusion Using two different approaches involving comparative sequence analyses, we could identify sequences from the early-diverging fungus B. emersonii previously considered specific to animals or plants, and highly divergent sequences from the same fungus relative to other fungi.

Changes in prolactin secretion in the short- and long-term after adrenalectomy

OBJECTIVE: To evaluate the modulation of the hypothalamus-pituitary-adrenal axis (HPA) on prolactin secretion in rats after adrenalectomy (ADX). MATERIALS AND METHODS: Plasma corticosterone, ACTH, and prolactin concentrations were measured by radioimmunoassay in rats after bilateral ADX in the short- (3 hours and 1day) and long-term (3, 7, and 14 days). RESULTS: Animals that underwent ADX showed undetectable corticosterone levels and a triphasic ACTH response with a transient increase (3h), a decrease (1d), and further increase in the long-term after ADX. Sham animals showed a marked increase in corticosterone and ACTH levels three hours after surgery, with a decrease to basal levels thereafter. Plasma prolactin levels were not changed after ADX. CONCLUSION: There are different points of equilibrium in the HPA axis after the glucocorticoid negative feedback is removed. Prolactin plasma secretion is not altered in the short or long- term after ADX, suggesting that the peptidergic neurons essential for prolactin release are not activated after ADX.

Panic-like defensive behavior but not fear-induced antinociception is differently organized by dorsomedial and posterior hypothalamic nuclei of Rattus norvegicus (Rodentia, Muridae)

The hypothalamus is a forebrain structure critically involved in the organization of defensive responses to aversive stimuli. Gamma-aminobutyric acid (GABA)ergic dysfunction in dorsomedial and posterior hypothalamic nuclei is implicated in the origin of panic-like defensive behavior, as well as in pain modulation. The present study was conducted to test the difference between these two hypothalamic nuclei regarding defensive and antinociceptive mechanisms. Thus, the GABA A antagonist bicuculline (40 ng/0.2 µL) or saline (0.9% NaCl) was microinjected into the dorsomedial or posterior hypothalamus in independent groups. Innate fear-induced responses characterized by defensive attention, defensive immobility and elaborate escape behavior were evoked by hypothalamic blockade of GABA A receptors. Fear-induced defensive behavior organized by the posterior hypothalamus was more intense than that organized by dorsomedial hypothalamic nuclei. Escape behavior elicited by GABA A receptor blockade in both the dorsomedial and posterior hypothalamus was followed by an increase in nociceptive threshold. Interestingly, there was no difference in the intensity or in the duration of fear-induced antinociception shown by each hypothalamic division presently investigated. The present study showed that GABAergic dysfunction in nuclei of both the dorsomedial and posterior hypothalamus elicit panic attack-like defensive responses followed by fear-induced antinociception, although the innate fear-induced behavior originates differently in the posterior hypothalamus in comparison to the activity of medial hypothalamic subdivisions.

Método de ensaio biomecânico para análise da isometricidade na reconstrução do ligamento patelofemoral medial

OBJETIVO: Apresentar um dispositivo biomecânico para o estudo da reconstrução do ligamento patelofemoral medial (LPFM) e sua isometricidade. MÉTODOS: Foi desenvolvido um sistema biomecânico acessível, que permite a aplicação de forças fisiológicas e não fisiológicas no joelho, através de um braço mecânico e aplicação de pesos e contrapesos, possibilitando a execução de diferentes estudos, além de ter um sistema de medidas bastante preciso de aferição de distâncias entre diferentes estruturas para análise dos experimentos. Este artigo descreve a montagem deste sistema, além de sugerir algumas aplicações práticas. Foram estudados seis joelhos de cadáveres. Os joelhos foram preparados em uma máquina de ensaios desenvolvida no Laboratório de Biomecânica do IOT HC FMUSP, que permitiu a avaliação dinâmica do comportamento patelar, quantificando a sua lateralização entre 0 e 120 graus. A diferença entre as distâncias encontradas, com e sem carga, aplicada na patela foram agrupadas segundo o ângulo de fixação do enxerto (0°, 30°, 60° e 90°) e situação do joelho (íntegro, reconstruído e lesado). RESULTADOS: Houve uma tendência em ocorrer menor desvio lateral em ângulos de fixação acima de 30 graus de flexão, principalmente entre os ângulos entre 45° e 60° graus de flexão, após a reconstrução. Para os demais ângulos não houve significância estatística. CONCLUSÃO: O método desenvolvido é uma ferramenta útil para os estudos da articulação patelofemoral, além de ter um sistema de medidas bastante preciso de aferição de distâncias entre diferentes estruturas e permitir a sua utilização em instituições com menos recursos disponíveis.

Estudo anatômico e funcional do complexo ligamentar colateral medial do cotovelo

OBJETIVO: Realizar um estudo anatômico do ligamento colateral medial, um importante estabilizador do cotovelo, em diferentes graus de flexo-extensão do cotovelo. MÉTODOS: Foram dissecados 40 cotovelos para analisar o comportamento funcional das bandas anterior, posterior e transversa do ligamento nas manobras de estresse em valgo do cotovelo durante seu movimento de flexão e extensão em diferentes graus. Determinou-se dois grupos: no GPA foi seccionado, a banda posterior do ligamento, depois a cápsula articular e, por fim, a banda anterior; no GAP, a ordem de dissecação foi inversa. RESULTADOS: No GPA observou-se instabilidade somente na terceira etapa e a média de abertura foi maior entre 50&º e 70&º de flexão de cotovelo; no GAP, a instabilidade apareceu desde a primeira etapa e os graus de flexão com maior instabilidade foram nos mesmos do grupo A. CONCLUSÃO: A banda anterior do ligamento colateral medial do cotovelo é o mais importante estabilizador na instabilidade em valgo do cotovelo e sua atuação principal acontece entre 50º e 70º de flexão do cotovelo. Nível de evidência III, Estudo Diagnóstico - Investigação de um exame para diagnóstico.

Análise do arco longitudinal medial em adolescentes usuárias de calçados de salto alto

O objetivo do estudo foi analisar a influência do calçado de salto alto no arco longitudinal medial (ALM) do pé de adolescentes. Fizeram parte do estudo 82 adolescentes entre 13 e 20 anos, sendo 54 não usuárias (grupo controle - GC) e 28 usuárias (grupo experimental - GE) de calçado de salto alto. Foram obtidas as impressões plantares de ambos os pés para análise do ALM, antes e depois do uso de um calçado de salto alto padronizado por um período de 30 minutos. As impressões plantares foram avaliadas pelo índice de Chippaux-Smirak (ICS) e pelo arco de Cavanagh & Rodgers (ICR). O teste de Shapiro-Wilks foi utilizado para a verificação da normalidade dos dados. Variáveis paramétricas pareadas foram tratadas com o Teste t de Student pareado e as não-paramétricas com o teste de Wilcoxon. As comparações não-pareadas foram realizadas com o teste t de Student para as variáveis paramétricas e o de Mann-Withney para as não-paramétricas, com nível de significância de 0,05. Houve diferença no ALM entre os lados direito e esquerdo apenas no GC antes do uso do calçado. Na comparação entre antes e depois do uso do sapato, notou-se diferença apenas no pé esquerdo do GC pelo ICS. Já entre GC e GE, não houve diferença. Apesar dos resultados não evidenciarem alterações no ALM, deve-se lembrar que esta é uma medida estática, sendo necessários estudos do componente dinâmico e do uso do calçado de salto crônico para correlacionar com os achados deste trabalho.

Ventral premammillary nucleus as a critical sensory relay to the maternal aggression network

Maternal aggression is under the control of a wide variety of factors that prime the females for aggression or trigger the aggressive event. Maternal attacks are triggered by the perception of sensory cues from the intruder, and here we have identified a site in the hypothalamus of lactating rats that is highly responsive to the male intruder—the ventral premammillary nucleus (PMv). The PMv is heavily targeted by the medial amygdalar nucleus, and we used lesion and immediate-early gene studies to test our working hypothesis that the PMv signals the presence of a male intruder and transfers this information to the network organizing maternal aggression. PMv-lesioned dams exhibit significantly reduced maternal aggression, without affecting maternal care. The Fos analysis revealed that PMv influences the activation of hypothalamic and septal sites shown to be mobilized during maternal aggression, including the medial preoptic nucleus (likely to represent an important locus to integrate priming stimuli critical for maternal aggression), the caudal two-thirds of the hypothalamic attack area (comprising the ventrolateral part of the ventromedial hypothalamic nucleus and the adjacent tuberal region of the lateral hypothalamic area, critical for the expression of maternal aggression), and the ventral part of the anterior bed nuclei of the stria terminalis (presently discussed as being involved in controlling neuroendocrine and autonomic responses accompanying maternal aggression). These findings reveal an important role for the PMv in detecting the male intruder and how this nucleus modulates the network controlling maternal aggression.

Temporal changes of CB1 cannabinoid receptor in the basal ganglia as a possible structure-specific plasticity process in 6-OHDA lesioned rats

The endocannabinoid system has been implicated in several neurobiological processes, including neurodegeneration, neuroprotection and neuronal plasticity. The CB1 cannabinoid receptors are abundantly expressed in the basal ganglia, the circuitry that is mostly affected in Parkinson’s Disease (PD). Some studies show variation of CB1 expression in basal ganglia in different animal models of PD, however the results are quite controversial, due to the differences in the procedures employed to induce the parkinsonism and the periods analyzed after the lesion. The present study evaluated the CB1 expression in four basal ganglia structures, namely striatum, external globus pallidus (EGP), internal globus pallidus (IGP) and substantia nigra pars reticulata (SNpr) of rats 1, 5, 10, 20, and 60 days after unilateral intrastriatal 6-hydroxydopamine injections, that causes retrograde dopaminergic degeneration. We also investigated tyrosine hydroxylase (TH), parvalbumin, calbindin and glutamic acid decarboxylase (GAD) expression to verify the status of dopaminergic and GABAergic systems. We observed a structure-specific modulation of CB1 expression at different periods after lesions. In general, there were no changes in the striatum, decreased CB1 in IGP and SNpr and increased CB1 in EGP, but this increase was not sustained over time. No changes in GAD and parvalbumin expression were observed in basal ganglia, whereas TH levels were decreased and the calbindin increased in striatum in short periods after lesion. We believe that the structure-specific variation of CB1 in basal ganglia in the 6-hydroxydopamine PD model could be related to a compensatory process involving the GABAergic transmission, which is impaired due to the lack of dopamine. Our data, therefore, suggest that the changes of CB1 and calbindin expression may represent a plasticity process in this PD model

Oral leucine supplementation is sensed by the brain but neither reduces food intake nor induces an anorectic pattern of gene expression in the hypothalamus

Leucine activates the intracellular mammalian target of the rapamycin (mTOR) pathway, and hypothalamic mTOR signaling regulates food intake. Although central infusion of leucine reduces food intake, it is still uncertain whether oral leucine supplementation is able to affect the hypothalamic circuits that control energy balance. We observed increased phosphorylation of p70s6k in the mouse hypothalamus after an acute oral gavage of leucine. We then assessed whether acute oral gavage of leucine induces the activation of neurons in several hypothalamic nuclei and in the brainstem. Leucine did not induce the expression of Fos in hypothalamic nuclei, but it increased the number of Fos-immunoreactive neurons in the area postrema. In addition, oral gavage of leucine acutely increased the 24 h food intake of mice. Nonetheless, chronic leucine supplementation in the drinking water did not change the food intake and the weight gain of ob/ob mice and of wild-type mice consuming a low- or a high-fat diet. We assessed the hypothalamic gene expression and observed that leucine supplementation increased the expression of enzymes (BCAT1, BCAT2 and BCKDK) that metabolize branched-chain amino acids. Despite these effects, leucine supplementation did not induce an anorectic pattern of gene expression in the hypothalamus. In conclusion, our data show that the brain is able to sense oral leucine intake. However, the food intake is not modified by chronic oral leucine supplementation. These results question the possible efficacy of leucine supplementation as an appetite suppressant to treat obesity