Impaired post exercise heart rate recovery in anabolic steroid users.

Previous study showed that muscle sympathetic nerve activity (MSNA) was augmented in anabolic steroids users (AASU). In the present study, we tested the hypothesis that the heart rate (HR) responses after maximal exercise testing would be reduced in AASU. 10 male AASU and 10 AAS nonusers (AASNU) were studied. Cardiopulmonary exercise was performed to assess the functional capacity and heart rate recovery. MSNA was recorded directly from the peroneal nerve by microneurography technique. Peak oxygen consumption (VO2) was lower in AASU compared to AASNU (43.66±2.24 vs. 52.70±1.68 ml/kg/min, P=0.005). HR recovery (HRR) at first and second minute was lower in AASU than AASNU (21±2 vs. 27±2 bpm, P=0.02 and 37±4 vs. 45±2 bpm, P=0.05, respectively). MSNA was higher in AASU than AASNU (29±3 vs. 20±1 bursts/min, P=0.01). Further analysis showed a correlation between HRR and MSNA (r=- 0.64, P=0.02), HRR at first minute and peak VO2 (r=0.70, P=0.01) and HRR at second minute and peak VO2 (r=0.62, P=0.02). The exacerbated sympathetic outflow associated with a lower parasympathetic activation after maximal exercise, which impairs heart rate recovery, strengthens the idea of autonomic imbalance in AASU.

Beta-sarcoglycanopathy (LGMD 2E) in a Spanish family.

Out of 10 autosomal recessive limb-girdle muscular dystrophies reported, 4 are caused by mutations in the genes encoding for sarcoglycans (alpha-, beta-, gamma- and delta-SG). Beta-sarcoglycanopathy (limb-girdle muscular dystrophy 2E) is a genetically heterogeneous disorder which usually presents a severe progressive clinical course. A complete immunohistochemical evaluation of the sarcoglycan complex should be carried out to direct the mutation analysis approach. The present report concerns a Spanish family with a genetically confirmed beta-sarcoglycanopathy. The patient, a 16-year-old female, offspring of a consanguineous marriage, developed a severe limb-girdle muscular dystrophy with a Duchenne-like phenotype. Muscle biopsy showed dystrophic changes and complete absence of the four sarcoglycans. Genetic analysis demonstrated homozygosis for the M100K missense mutation in exon 3, encoding for the proximal extracellular domain. The parents and one sister were found to be carriers. Missense mutations affecting this domain result in the instability of the entire sarcoglycan complex and lead to severe phenotypes as seen in non-sense mutations.

Molecular diagnosis and counseling in a family presenting compound heterozygosity for autosomal recessive limb-girdle muscular dystrophy.

The present report concerns two patients, male and female siblings, manifesting a different degree of severity for the same autosomal recessive limb-girdle muscular dystrophy. The index case (male sib) carried the clinical diagnosis of Becker muscular dystrophy at the time when the sister, with a much milder presentation, first sought counseling and prenatal diagnosis for a pregnancy already in course. Molecular and immunocytochemical tests then available favoured the diagnosis of an autosomal recessive myopathy, but did not enable exclusion of a dystrophinopathy The couple was counseled accordingly, although prenatal diagnosis could not be offered. Both patients were later found to carry one gamma- and two alpha-sarcoglycan gene mutations, one of the latter being new This raised a counseling dilemma: depending on which combination was the disease-causing genotype, there would be a minimal or a significant 25% risk to offspring. We describe the studies carried out and emphasise the importance of differential diagnosis and extensive molecular characterisation in this group of disorders, so as to enable correct genetic counseling and prenatal diagnosis.

A capture enzyme-linked immunosorbent assay for species-specific detection of Bothrops venoms.

A direct sandwich enzyme-linked immunosorbent assay (ELISA), employing affinity purified antivenom antibodies specifically recognizing the homologous venom, was developed for species-specific detection of bothropic venom. The method is based on a two-step affinity purification of the specific antibodies. A species monovalent antivenom is adsorbed onto a venom adsorbent containing heterologous venoms from the Bothrops, Crotalus and Lachesis genera. The species-specific antibodies obtained, are then adsorbed onto a second venom adsorbent containing only the homologous venom for the removal of non antivenom antibodies. Venom concentrations of 0.1 and 1,000 ng/ml were specifically identified for Bothrops jararacussu and B. alternatus venom respectively.

Analysis of FMR1 and flanking microsatellite markers in normal and fragile X chromosomes in Portugal: evidence for a "protector" haplotype.

In order to look for linkage disequilibrium between the fragile X locus and its flanking markers, we analysed the FRAXAC1 and DXS548 microsatellites in normal and fragile X individuals of Portuguese origin. We observed differences in allele and haplotype frequencies between these two samples. Four haplotypes (A-2, C-2, C-5 and D-6) accounted for 76% of all fragile X chromosomes, whereas a single haplotype (C-7) accounted for 70% of the normal population and less than 3% of the fragile X chromosomes. Among the four observed high-risk haplotypes, A-2 and D-6 had been previously reported in other studies, but C-2 and C-5 seem characteristic of Portuguese patients, as suggested by the high frequency (38%) in fragile X chromosomes and virtual absence in controls. In accordance with previous studies, a greater heterozygosity of the fragile X sample was noted when compared to that of controls. The high frequency of C-7 haplotype in the normal population and its virtual absence in the fragile X sample may reflect the existence of linkage disequilibrium between the two loci and/or selective advantage (protector effect) of this haplotype.

Development of an ELISA to assess the potency of horse therapeutic polyvalent antibothropic antivenom.

The objective of this study was the search for a suitable venom antigen to be used in an in vitro alternative immunoassay, to the standard antivenom neutralization assay using mice. Bothrops jararaca venom was fractionated in DEAE-Sephacel columns and the fractions were tested for a correlation between antibody capture enzyme linked immunosorbent assay (ELISA) absorbance values and the 'in vivo' antivenom potency. Individual antivenoms from 14 horses and 15 separate FUNED polyspecific Bothrops ampouled antivenoms (final product) were used. Fractions showing the higher correlations were further chromatographed in a Sephadex G-75 column and again tested for the correlation. Two fractions with haemorrhagic activity displayed a correlation of r = 0.77 and r = 0.8 against the individual horse antivenom sera and of r = 0.79 and r = 0.8 for the ampouled antivenom. For all results p < 0.001. Two other fractions with phospholipase A2 activity showed a correlation of r = 0.66 (p < 0.01) and r = 0.56 (p < 0.03) against the individual horse antivenom sera. Electrophoresis results show a similar composition for both antigens with haemorrhagic activity. Results indicate that the fractions purified would be suitable for the desired objective of this study.

Noval mutation (Y184C) in exon 4 of the beta-sarcoglycan gene identified in a Portuguese patient. Mutations in brief no. 177. Online.

We report a novel beta-sarcoglycan gene mutation identified in a 21-year-old Portuguese male with a progressive myopathy of intermediate severity, who had been misdiagnosed as Becker Muscular Dystrophy (BMD) based on clinical observations and muscle immunocytochemical anaylsis with dystrophin antibodies only. Since no detectable deletions or duplications were found in the dystrophin gene, we screened for mutations in the sarcoglycan genes by PCR-SSCP. The patient's sample showed a band of increased mobility in exon 4 of the beta-sarcoglycan gene which, upon sequencing, was found to represent a homozygous A-->G transversion at nucleotide 551, resulting in a tyrosine to cysteine substitution at position 184 (Y184C). Carrier status was ascertained in both parents and a sister. These aberrant conformers were not detected in 85 unrelated control individuals screened by PCR-SSCP analysis. All seven beta-sarcoglycan mutations reported to date are associated with a severe phenotype and occur in exons 3 and 4, which correspond to the immediate extracellular domain of the protein. This region contains five conserved cysteine residues. In our patient, the presence of an extra cysteine residue could interefere with intra- and/or inter-molecular disulphide bond formation. The intermediate phenotype could perhaps result from the assembly of both normal and abnormal complexes, depending on the formation of the disulphide bonds.

[Leptospirosis in animal reproduction: III. Role of the hardjo serovar in bovine leptospirosis in Rio de Janeiro, Brazil]. / Leptospirose em reprodução animal: III. Papel do serovar hardjo nas leptospiroses bovinas no Rio de Janeiro, Brasil.

Four hundred and five serum samples were drawn from cows with reproductive problems which were not vaccinated against leptospirosis from 21 dairy farms. Three distinct geographic regions were determined and the farms were also classified considering the production system, based on technological, zootechnical and sanitary resources. A total of 277 positive reactions were observed, corresponding to 68.39% of the samples. The predominant serovar was hardjo, reactive on 85 samples (20.98%), predominant on nine farms and observed on 17 farms (80.95%). It was observed the predominance of hardjo in all studied regions and on properties classified as type "A" (22 samples) and type "B" (49 samples). The role of this serovar on bovine leptospirosis in Brazil compared with other countries is discussed.

GM2-gangliosidosis B1 variant: analysis of beta-hexosaminidase alpha gene mutations in 11 patients from a defined region in Portugal.

The GM2-gangliosidosis B1 variant occurs at an exceptionally high frequency in the northern part of Portugal. In most patients, the disease manifests itself as a juvenile form, as opposed to the late-infantile form described for many patients from other parts of the world. We have analyzed the beta-hexosaminidase alpha gene in 11 patients, as well as in some relatives, in order to characterize the underlying abnormalities. They were screened for the two previously identified mutations responsible for the B1 variant phenotype (G533----A, also designated as the "DN allele," and C532---T) by PCR amplification of an 800-bp DNA fragment and subsequent dot-blot hybridization with allele-specific oligonucleotides. The fragment amplified from one patient was also subcloned and sequenced. Ten patients, constituting a clinically and biochemically homogeneous group, were found to be homozygous for the DN allele. The other, whose clinical profile more resembled the late-infantile phenotype often described in the literature, was a compound heterozygote carrying the DN allele and another, as yet unidentified, abnormal allele. Our results, corroborated by previously published data, suggest that homozygotes and compound heterozygotes for the DN allele may be distinguishable at the phenotypic level, depending on the nature of the abnormality in the other allele. A common ancestral origin for the DN allele can also be postulated.

Biochemical characterization of beta-hexosaminidase in different biological specimens from eleven patients with GM2-gangliosidosis B1 variant.

GM2-gangliosidosis B1 variant is thought to be a rare disorder with a wide geographical and ethnic distribution. We report the biochemical findings obtained in different specimens from a group of 11 B1 variant patients originating from the north of Portugal. The biochemical data obtained seem to indicate that only one of these patients is a genetic compound presenting a clinical and biochemical pattern similar to the majority of B1 variant patients described in the literature, but somewhat different from the profile presented by the other patients reported here, who are homozygous for the 'DN-allele'.

Censo Epidemiológico de lesôes e afeccôes cutâneas em escolares de Ribeirâo Prêto, S. P., Brasil / Epidemiologieal census of cutaneous lesions in schoolchildren of R. Preto, S.P., Brazil

Um total de 5150 escolares de 6 a 16 anos foram examinados em uma cidade subtropical. Excluídas cicatrizes e lesôes traumáticas a prevalência de dermatoses foi de 72%. As mais comun (>2%) foram nevos pigmentados (72%),cicatrizes e lesôes residuais (36%), efélides (19,6%), pediculose (19,6%), dermatite seborréica (19,5%), dermatoviroses (18,6%), acne (15,7%), lesôes traumáticas (9,5%), pitiríase capitis (5%), piodermites (4,3%), dermatite eozematosa (2,6%), estrófulo + picada de inseto (3,1%), ceratose palmar el ou plantar (2,1%) e disidrose (2%). Muitos doentes apresentaron dois ou mais tipos de lesoes ou affeccôes cutâneas. A prevalencia de vibix, estomatite angular, dermatoses actínicas, ictiose, pele xerótica, intertrigo, milia, livedo reticular, hipertricosee escabiose foi menor porén relativamente elevada (de 1,61 a 0,66%). A prevalencia foi discretamente maior em escolares de sexo masculino que no feminino e as diferencas estatisticamente significantes (p<0,001). A diferenca de prevalência entre os grupos etarios nâo foi significante para masculinos e femininos. As meninas aprentaron taxa mais elevada de pediculose, enquanto em meninos prevaleceram pitiríase alba (dartro volante), pitiríase versicolor e piodermites. As condicôes sócioeconômicas parecen ter tido influencia nas taxas de prevalênia de pediculose, pitiríase alba e "tinea pedis", mas nâo nas de dermatoviroses, pitiríase versicolor e piodermite. Os resultados e comentários sâo válidos para uma àrea subtropical semelhante à de Ribeirâo Prêto, com as mesmas características ecológicas, socio-econômicas e de populacâo. Quando se planeja um programa de saúde pública em escolas é importante realizar antes um censo epidemiológico ou estudo piloto, a fim de obter as relevantes informacôes bàsicas.

Censo Epidemiológico de les¶es e afecc¶es cutÔneas em escolares de RibeirÔo PrÛto, S. P., Brasil / Epidemiologieal census of cutaneous lesions in schoolchildren of R. Preto, S.P., Brazil

Um total de 5150 escolares de 6 a 16 anos foram examinados em uma cidade subtropical. Excluídas cicatrizes e les¶es traumáticas a prevalÛncia de dermatoses foi de 72%. As mais comun (>2%) foram nevos pigmentados (72%),cicatrizes e les¶es residuais (36%), efélides (19,6%), pediculose (19,6%), dermatite seborréica (19,5%), dermatoviroses (18,6%), acne (15,7%), les¶es traumáticas (9,5%), pitiríase capitis (5%), piodermites (4,3%), dermatite eozematosa (2,6%), estrófulo + picada de inseto (3,1%), ceratose palmar el ou plantar (2,1%) e disidrose (2%). Muitos doentes apresentaron dois ou mais tipos de lesoes ou affecc¶es cutÔneas. A prevalencia de vibix, estomatite angular, dermatoses actínicas, ictiose, pele xerótica, intertrigo, milia, livedo reticular, hipertricosee escabiose foi menor porén relativamente elevada (de 1,61 a 0,66%). A prevalencia foi discretamente maior em escolares de sexo masculino que no feminino e as diferencas estatisticamente significantes (p<0,001). A diferenca de prevalÛncia entre os grupos etarios nÔo foi significante para masculinos e femininos. As meninas aprentaron taxa mais elevada de pediculose, enquanto em meninos prevaleceram pitiríase alba (dartro volante), pitiríase versicolor e piodermites. As condic¶es sócioecon¶micas parecen ter tido influencia nas taxas de prevalÛnia de pediculose, pitiríase alba e "tinea pedis", mas nÔo nas de dermatoviroses, pitiríase versicolor e piodermite. Os resultados e comentários sÔo válidos para uma Orea subtropical semelhante O de RibeirÔo PrÛto, com as mesmas características ecológicas, socio-econ¶micas e de populacÔo. Quando se planeja um programa de saúde pública em escolas é importante realizar antes um censo epidemiológico ou estudo piloto, a fim de obter as relevantes informac¶es bOsicas. (AU)

Depressed cell-mediated immunity in iron-deficiency anemia due to chronic loss of blood.

Twelve women with iron-deficiency anemia due to chronic loss of blood, but free from any other pathology that might alter the immune response, were studied. The patients were tested for cell immunity both in vitro, by B and T lymphocyte quantitation and by blastic transformation of the lymphocytes with phytohemagglutinin (PHA), and in vivo, by dinitrochlorobenzene (DNCB), tuberculin, trychophytine and varidase skin tests. The same tests were repeated after iron therapy and also applied to a group of 12 normal control subjects. The percent of T lymphocytes increased significantly from 55.1 to 66.0% after treatment, while the absolute values did not change. There was a significant decrease in both the number and percent of "null" lymphocytes after treatment. The percent and absolute number of B lymphocytes did not change after treatment. Blastic transformation indices were within the normal range both before and after treatment. Seven women who were DNBC-negative before treatment became DNBC-positive after iron therapy. Of the 5 patients who were tuberculin-negative before treatment, 2 became positive after iron therapy. Reactivity to trychophytine was observed in 3 patients before treatment as compared to 5 afterwards. Reactivity to varidase increased from 4 to 6 patients upon iron therapy. On the basis of changes in immunological reactivity observed after iron replenishment, we conclude that iron deficiency is an important factor in the genesis of the immunological alterations occurring in iron-deficiency anemia.