Hereditary motor and autonomic neuronopathy 1 maps to chromosome 20q13.2-13.3

The spinal muscular atrophies (SMA) or hereditary motor neuronopathies result from the continuous degeneration and death of spinal cord lower motor neurons, leading to progressive muscular weakness and atrophy. We describe a large Brazilian family exhibiting an extremely rare, late-onset, dominant, proximal, and progressive SMA accompanied by very unusual manifestations, such as an abnormal sweating pattern, and gastrointestinal and sexual dysfunctions, suggesting concomitant involvement of the autonomic nervous system. We propose a new disease category for this disorder, `hereditary motor and autonomic neuronopathy', and attribute the term, `survival of motor and autonomic neurons 1' (SMAN1) to the respective locus that was mapped to a 14.5 cM region on chromosome 20q13.2-13.3 by genetic linkage analysis and haplotype studies using microsatellite polymorphic markers. This locus lies between markers D20S120 and D20S173 showing a maximum LOD score of 4.6 at D20S171, defining a region with 33 known genes, including several potential candidates. Identifying the SMAN1 gene should not only improve our understanding of the molecular mechanisms underlying lower motor neuron diseases but also help to clarify the relationship between motor and autonomic neurons.

Protein defects in neuromuscular diseases

Muscular dystrophies are a heterogeneous group of genetically determined progressive disorders of the muscle with a primary or predominant involvement of the pelvic or shoulder girdle musculature. The clinical course is highly variable, ranging from severe congenital forms with rapid progression to milder forms with later onset and a slower course. In recent years, several proteins from the sarcolemmal muscle membrane (dystrophin, sarcoglycans, dysferlin, caveolin-3), from the extracellular matrix (alpha2-laminin, collagen VI), from the sarcomere (telethonin, myotilin, titin, nebulin), from the muscle cytosol (calpain 3, TRIM32), from the nucleus (emerin, lamin A/C, survival motor neuron protein), and from the glycosylation pathway (fukutin, fukutin-related protein) have been identified. Mutations in their respective genes are responsible for different forms of neuromuscular diseases. Protein analysis using Western blotting or immunohistochemistry with specific antibodies is of the utmost importance for the differential diagnosis and elucidation of the physiopathology of each genetic disorder involved. Recent molecular studies have shown clinical inter- and intra-familial variability in several genetic disorders highlighting the importance of other factors in determining phenotypic expression and the role of possible modifying genes and protein interactions. Developmental studies can help elucidate the mechanism of normal muscle formation and thus muscle regeneration. In the last fifteen years, our research has focused on muscle protein expression, localization and possible interactions in patients affected by different forms of muscular dystrophies. The main objective of this review is to summarize the most recent findings in the field and our own contribution.

Restless leg syndrome, sleep quality and fatigue in multiple sclerosis patients

We have tested the hypothesis that restless leg syndrome (RLS) is related to quality of sleep, fatigue and clinical disability in multiple sclerosis (MS). The diagnosis of RLS used the four minimum criteria defined by the International Restless Legs Syndrome Study Group. Fatigue was assessed by the Fatigue Severity Scale (FSS >27), quality of sleep by the Pittsburgh Sleep Quality Index (PSQI >6), excessive daytime sleepiness by the Epworth Sleepiness Scale (ESS >10) and clinical disability by the Expanded Disability Status Scale (EDSS). Forty-four patients (32 women) aged 14 to 64 years (43 ± 14) with disease from 0.4 to 23 years (6.7 ± 5.9) were evaluated. Thirty-five were classified as relapsing-remitting, 5 as primary progressive and 4 as secondary progressive. EDSS varied from 0 to 8.0 (3.6 ± 2.0). RLS was detected in 12 cases (27%). Patients with RLS presented greater disability (P = 0.01), poorer sleep (P = 0.02) and greater levels of fatigue (P = 0.03). Impaired sleep was present in 23 (52%) and excessive daytime sleepiness in 3 cases (6.8%). Fatigue was present in 32 subjects (73%) and was associated with clinical disability (P = 0.000) and sleep quality (P = 0.002). Age, gender, disease duration, MS pattern, excessive daytime sleepiness and the presence of upper motor neuron signs were not associated with the presence of RLS. Fatigue was best explained by clinical disability and poor sleep quality. Awareness of RLS among health care professionals may contribute to improvement in MS management.

Morphological development of Corydoras aff. paleatus (Siluriformes, Callichthyidae) and correlation with the emergence of motor and social behaviors

Here we examine major anatomical characteristics of Corydoras aff. paleatus (Jenyns, 1842) post-hatching development, in parallel with its neurobehavioral evolution. Eleutheroembryonic phase, 4.3-8.8 days post-fertilization (dpf); 4.3-6.4 mm standard length (SL) encompasses from hatching to transition to exogenous feeding. Protopterygiolarval phase (8.9-10.9 dpf; 6.5-6.7 mm SL) goes from feeding transition to the commencement of unpaired fin differentiation, which marks the start of pterygiolarval phase (11-33 dpf; 6.8-10.7 mm SL) defined by appearance of lepidotrichia in the dorsal part of the median finfold. This phase ends with the full detachment and differentiation of unpaired fins, events signaling the commencement of the juvenile period (34-60 dpf; 10.8-18.0 mm SL). Eleutheroembryonic phase focuses on hiding and differentiation of mechanosensory, chemosensory and central neural systems, crucial for supplying the larval period with efficient escape and nutrient detection-capture neurocircuits. Protopterygiolarval priorities include visual development and respiratory, digestive and hydrodynamic efficiencies. Pterygiolarval priorities change towards higher swimming efficacy, including carangiform and vertical swimming, necessary for the high social interaction typical of this species. At the end of the protopterygiolarval phase, simple resting and foraging aggregations are seen. Resting and foraging shoals grow in complexity and participant number during pterygiolarval phase, but particularly during juvenile period.

Quantitative evidence for neurofilament heavy subunit aggregation in motor neurons of spinal cords of patients with amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS), a neurodegenerative disease of unknown etiology, affects motor neurons leading to atrophy of skeletal muscles, paralysis and death. There is evidence for the accumulation of neurofilaments (NF) in motor neurons of the spinal cord in ALS cases. NF are major structural elements of the neuronal cytoskeleton. They play an important role in cell architecture and differentiation and in the determination and maintenance of fiber caliber. They are composed of three different polypeptides: light (NF-L), medium (NF-M) and heavy (NF-H) subunits. In the present study, we performed a morphological and quantitative immunohistochemical analysis to evaluate the accumulation of NF and the presence of each subunit in control and ALS cases. Spinal cords from patients without neurological disease and from ALS patients were obtained at autopsy. In all ALS cases there was a marked loss of motor neurons, besides atrophic neurons and preserved neurons with cytoplasmic inclusions, and extensive gliosis. In control cases, the immunoreaction in the cytoplasm of neurons was weak for phosphorylated NF-H, strong for NF-M and weak for NF-L. In ALS cases, anterior horn neurons showed intense immunoreactivity in focal regions of neuronal perikarya for all subunits, although the difference in the integrated optical density was statistically significant only for NF-H. Furthermore, we also observed dilated axons (spheroids), which were immunopositive for NF-H but negative for NF-M and NF-L. In conclusion, we present qualitative and quantitative evidence of NF-H subunit accumulation in neuronal perikarya and spheroids, which suggests a possible role of this subunit in the pathogenesis of ALS.

Alguns aspectos epidemiológicos da mortalidade por acidentes de trânsito de veículo a motor na Cidade de São Paulo, Brasil

Foram estudadas algumas características dos óbitos por acidentes de trânsito de veículos a motor, no município de São Paulo, ocorridos entre 1.° de janeiro a 31 de dezembro de 1970. Todas as características do falecido e do acidente foram coletadas a partir dos dados registrados nos laudos de necrópsias existentes no Instituto Médico Legal. O estudo evidenciou que a mortalidade por acidentes de veículo a motor é alta, maior no sexo masculino, aumenta com a idade, sendo que o maior coeficiente foi para maiores de 60 anos. A zona da cidade com maior número de acidentes é a zona Sul, existindo áreas (distritos policiais) e vias públicas preferenciais quanto a ocorrência, em todas as 4 zonas do município; a maior ocorrência de acidentes foi aos sábados e domingos; os pedestres compreendem a grande maioria dos falecidos; proporção apreciável dos falecidos recebeu atendimento hospitalar após o acidente. Foram relacionados também o número total de acidentes, vítimas e mortes mostrando que para cada 100 acidentes ocorreram 62,50 vítimas e 5,13 mortes, e para cada 100 vítimas, 18, 22 mortes.

Aspectos epidemiológicos dos acidentes fatais a veículo a motor na cidade do Salvador (Bahia), Brasil

Foi realizado um estudo com o objetivo de descrever certos aspectos epidemiológicos dos acidentes de trânsito de veículo a motor, na cidade do Salvador, Bahia, Brasil, o qual envolveu coleta de informações no Departamento de Trânsito (DETRAN) e no Instituto de Medicina Legal. Ficou demonstrado que os acidentes de trânsito de veículo a motor são mais freqüentes ao fim da tarde e concentram-se, sobretudo, no fim de semana, isto é, sábado e domingo. Motoristas do sexo masculino foram muito mais freqüentemente envolvidos em acidentes de trânsito do que os do sexo feminino. Foi concluído que os acidentes fatais envolvendo pedestres constituem-se importante problema urbano necessitando cuidadosa atenção por parte de educadores e dos serviços públicos responsáveis pelo setor do trânsito. Foi sugerida a necessidade de melhorar e tornar mais uniforme o sistema de notificação e classificação dos acidentes de veículo a motor pelo DETRAN da cidade do Salvador.

Traumatismo crânio-encefálico: diferenças das vítimas pedestres e ocupantes de veículos a motor

OBJETIVO: Caracterizar as diferenças das vítimas com diagnóstico de traumatismo crânio-encefálico envolvidas em diferentes condições, em acidentes de trânsito de veículo a motor e evidenciar indicadores para prevenção e seu atendimento. MÉTODO: Foram analisados os prontuários de todas as vítimas com diagnóstico de trauma crânio-encefálico, assistidas em hospital de referência para atendimento do trauma, entre março e junho de 1993. A população foi de 156 vítimas, sendo 80 pedestres, 50 ocupantes de veículos a motor, exceto de moto e, 26 motociclistas ou passageiros de motos. RESULTADOS E CONCLUSÕES: Os resultados mostraram que a mortalidade entre pedestres foi a mais alta entre os três grupos (25,0%). Os óbitos ocorrem em 19,2% dos ocupantes de motocicleta e 8,0% dos ocupantes de demais veículos a motor. Foram observadas diferenças estatísticas entre os grupos quando a variável gravidade do trauma crânio-encefálico foi analisado. Análise de certas variáveis mostraram importantes diferenças na distribuição dos três grupos.

Oportunidades do ambiente domiciliar para o desenvolvimento motor

OBJETIVO: Avaliar as oportunidades presentes no ambiente domiciliar para o desenvolvimento motor de lactentes. MÉTODOS: Estudo epidemiológico de base populacional, transversal, realizado com 239 lactentes com idade entre três e 18 meses, residentes no município de Juiz de Fora, MG, em 2010. Os participantes foram selecionados por amostragem aleatória estratificada, conglomerada, em múltiplos estágios. Para avaliar a qualidade e quantidade de estímulo motor no ambiente domiciliar foi utilizado o instrumento Affordance in the Home Environment for Motor Development - Infant Scale. Procedeu-se a análise bivariada com aplicação do teste qui-quadrado, seguida de regressão logística multinomial para verificar a associação entre as oportunidades presentes no domicílio e fatores biológicos, comportamentais, demográficos e socioeconômicos. RESULTADOS: As oportunidades de estimulação ambiental foram relativamente baixas. Na análise bivariada, para a faixa etária de três a nove meses, foi encontrada associação com os fatores: ordem de nascimento (p = 0,06), classificação socioeconômica (p = 0,08), renda mensal (p = 0,06) e renda per capita (p = 0,03). No modelo de regressão, prevaleceu a classificação socioeconômica (RC = 7,46; p = 0,03). Para a faixa etária de dez a 18 meses, os fatores associados, na análise bivariada, foram: estado civil materno (p < 0,01), convívio da criança com o pai (p = 0,08), chefe da família (p = 0,04), número de pessoas no domicílio (p = 0,05), escolaridade materna (p < 0,01) e paterna (p < 0,01), classificação socioeconômica (p < 0,01) e renda per capita (p = 0,03). No modelo de regressão, o estado civil materno (RC = 4,83; p = 0,02), escolaridade materna (RC = 0,29; p = 0,03) e paterna (RC = 0,33; p = 0,04) permaneceram associados às oportunidades de estimulação ambiental. CONCLUSÕES: A união estável dos pais, maior escolaridade materna e paterna e maior nível econômico foram os fatores associados às melhores oportunidades de estimulação motora no lar.

Content validity and reliability of test of gross motor development in Chilean children

ABSTRACT OBJECTIVE To validate a Spanish version of the Test of Gross Motor Development (TGMD-2) for the Chilean population. METHODS Descriptive, transversal, non-experimental validity and reliability study. Four translators, three experts and 92 Chilean children, from five to 10 years, students from a primary school in Santiago, Chile, have participated. The Committee of Experts has carried out translation, back-translation and revision processes to determine the translinguistic equivalence and content validity of the test, using the content validity index in 2013. In addition, a pilot implementation was achieved to determine test reliability in Spanish, by using the intraclass correlation coefficient and Bland-Altman method. We evaluated whether the results presented significant differences by replacing the bat with a racket, using T-test. RESULTS We obtained a content validity index higher than 0.80 for language clarity and relevance of the TGMD-2 for children. There were significant differences in the object control subtest when comparing the results with bat and racket. The intraclass correlation coefficient for reliability inter-rater, intra-rater and test-retest reliability was greater than 0.80 in all cases. CONCLUSIONS The TGMD-2 has appropriate content validity to be applied in the Chilean population. The reliability of this test is within the appropriate parameters and its use could be recommended in this population after the establishment of normative data, setting a further precedent for the validation in other Latin American countries.

Differentiation of pathogenic and non-pathogenic leptospires by means of the polymerase chain reaction

A polymerase chain reaction was carried out to detect pathogenic leptospires isolated from animals and humans in Argentina. A double set of primers (G1/G2, B64-I/B64-II), described before, were used to amplify by PCR a DNA fragment from serogroups belonging to Leptospira interrogans but did not allow to detect saprophytic strains isolated from soil and water (L. biflexa). This fact represents an advantage since it makes possible the differentiation of pathogenic from non-pathogenic leptospires in cultures. The sensitivity of this assay has been determined, allowing to detect just only 10 leptospires in the reaction tube. Those sets of primers generated either a 285 bp or 360 bp fragment, depending on the pathogenic strain

Strain differentiation of Trichophyton rubrum by random amplification of polymorphic DNA (RAPD)

Trichophyton rubrum is an important cause of dermatomycoses. Molecular strain typing methods have recently been developed to address questions about epidemiology and source of relapse following treatment. This report describes the application of RAPD for molecular strain differentiation of this fungus utilizing the primers 1- (5'-d[GGTGCGGGAA]-3') and 6- (5'-d[CCCGTCAGCA]-3'). A total of five RAPD patterns were observed among 10 strains of T. rubrum, with each of the primers used. We conclude that RAPD analysis using primers 1 and 6 can be used in epidemiological studies.

Comparison among tomato juice agar with other three media for differentiation of Candida dubliniensis from Candida albicans

The purpose of the present study is to compare the tomato juice agar, a well known medium employed to observe ascospore formation, with niger seed agar, casein agar and sunflower seed agar, applied to a differentiation between C. dubliniensis and C. albicans. After 48 hours of incubation at 30 ºC all 26 (100%) C. dubliniensis isolates tested produced chlamydospores on tomato juice agar as well as in the other three media evaluated. However, when we inoculated all media with C. albicans, the absence of chlamydospores became resulting in the following percents: tomato juice agar (92.47%), niger seed agar (96.7%), casein agar (91.39%), and sunflower seed agar (96.7%). These results indicate that tomato juice agar is another medium which can also be used in the first phenotypic differentiation between C. dubliniensis and C. albicans.

Differentiation of Candida dubliniensis from Candida albicans with the use of killer toxins

The aim of this study was to report the ability of killer toxins, previously used as biotyping techniques, as a new tool to differentiate C. albicans from C. dubliniensis. The susceptibility of C. albicans and C. dubliniensis to killer toxins ranged from 33.9 to 93.3% and from 6.67 to 93.3%, respectively.

MULTIPLEX SYBR® GREEN-REAL TIME PCR (qPCR) ASSAY FOR THE DETECTION AND DIFFERENTIATION OF Bartonella henselae AND Bartonella clarridgeiae IN CATS

A novel SYBR® green-real time polymerase chain reaction (qPCR) was developed to detect two Bartonellaspecies, B. henselae and B. clarridgeiae, directly from blood samples. The test was used in blood samples obtained from cats living in animal shelters in Southern Brazil. Results were compared with those obtained by conventional PCR targeting Bartonella spp. Among the 47 samples analyzed, eight were positive using the conventional PCR and 12 were positive using qPCR. Importantly, the new qPCR detected the presence of both B. henselae and B. clarridgeiae in two samples. The results show that the qPCR described here may be a reliable tool for the screening and differentiation of two important Bartonella species.