Determination of testicular function in adolescents with varicocoele - a proteomics approach.

The goal of this study was to determine seminal plasma biomarkers of testicular function in adolescents with varicocoele and to verify enriched gene ontology terms associated to these differential proteomes. An observational study was carried out in an academic research environment. A total of 77 adolescent patients were recruited from a local public school, of which 23 were without varicocoele and with normal semen analysis (control group), 37 were with varicocoele and normal semen (VNS) parameters, and 17 were with varicocoele and altered semen (VAS) parameters. Two semen collections were provided with a 1-week interval, after 2-5 days of ejaculatory abstinence. Seminal plasma proteins were identified and quantified utilizing a label-free shotgun proteomics approach, generating (i) proteins differentially expressed in each group (control, VNS, and VAS) and putative biomarkers using multivariate statistics followed by discriminant analysis. Confirmatory analysis was performed for two proteins by western blotting. Enriched biological processes and molecular functions were determined using gene ontology analysis. In total, 541 proteins were identified and quantified: 108 exclusive or overexpressed in controls, 26 in the VNS group, and 13 in the VAS group. The suggested biomarkers are Cab45/SDF4 (Q9BRK5), protein lefty-1 (O75610), DNase I (P24855), PAP2-alpha (O14494), IBP-7 (Q16270), HDC (P01860), and CRISP-3 (P54108). Western blotting results showed that Cab45 was significantly underexpressed in both varicocoele groups, and CRISP-3 was significantly overexpressed in seminal plasma of adolescents with VAS. In conclusion, specific biomarkers of spermatogenesis and homeostasis are observed in adolescents without varicocoele, and the presence of a palpable varicocoele progressively shifts these adolescents toward initially an immune response, and finally toward a chronic inflammatory profile. This shift is accompanied by decreased semen quality.

Unbiased label-free quantitative proteomic profiling and enriched proteomic pathways in seminal plasma of adult men before and after varicocelectomy.

STUDY QUESTION: Does the seminal plasma proteomic profile and functional enrichment of gene ontology terms change after microsurgical varicocelectomy? Are there any potential targets for diagnosis or therapeutic intervention in varicocele? SUMMARY ANSWER: A shift in state from a responsive-to-stress condition before varicocele correction to a responsive-to-environment condition after varicocelectomy was observed in enriched proteomic pathways. WHAT IS KNOWN ALREADY: Varicocele may lead to many adverse effects, including failure of testicular growth and development, and is associated with decreased semen quality and increased semen oxidative stress. Varicocelectomy is the treatment of choice, and is associated with improved semen quality, but little is known regarding the underlying molecular mechanisms and post-genomic pathways following intervention. STUDY DESIGN, SIZE, DURATION: A prospective study was carried out including 18 adult men with varicocele. These patients provided one semen sample before they were submitted for bilateral varicocele repair through microsurgical varicocelectomy, and one other semen sample 90 days after the surgery. PARTICIPANTS/MATERIALS, SETTING, METHODS: An aliquot of each semen sample was used for unbiased proteomics analysis by a label-free quantitative approach (2D nanoUPLC-ESI-MS(E)). Samples were pooled according to group (normalized to protein content) and run in quadruplicate. These quadruplicate runs provided degrees of freedom in order to compare groups using a non-parametric Mann-Whitney test for quantified proteins. MAIN RESULTS AND THE ROLE OF CHANCE: A total of 316 proteins were quantified or identified, of which 91 were exclusively identified or quantified in one of the groups (53 in the pre- and 38 in the post-varicocelectomy group), and 68 were quantified in both groups and submitted to statistical analysis, of which 5 were overrepresented in the pre-varicocelectomy group (P < 0.05). In enriched functional analysis, binding and response to stimulus functions were enriched in a common cluster (present in both groups), nitric oxide metabolism and tetratricopeptide repeat domain-binding functions were enriched in the pre-varicocelectomy group, and response to reactive oxygen species, gluconeogenesis, nicotinamide adenine dinucleotide-binding and protein stabilization were enriched in the post-varicocelectomy. LIMITATIONS, REASONS FOR CAUTION: Because a shotgun proteomics analysis was chosen in order to generate a list of putative biomarkers, a targeted follow-up study should be performed to confirm these biomarkers. WIDER IMPLICATIONS OF THE FINDINGS: The proteins found in both groups possess functions usually found in human semen. The enriched function analysis demonstrated a shift back to homeostasis after varicocelectomy, suggesting that varicocele correction promotes return of semen to a physiological state. STUDY FUNDING/COMPETING INTEREST(S): The funding for this project was received from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) as a scholarship for Ms Camargo. There was no conflict of interest.

Genetic polymorphisms modulate the folate metabolism of Brazilian individuals with Down syndrome.

Individuals with Down syndrome (DS) carry three copies of the Cystathionine ß-synthase (CßS) gene. The increase in the dosage of this gene results in an altered profile of metabolites involved in the folate pathway, including reduced homocysteine (Hcy), methionine, S-adenosylhomocysteine (SAH) and S-adenosylmethionine (SAM). Furthermore, previous studies in individuals with DS have shown that genetic variants in genes involved in the folate pathway influence the concentrations of this metabolism's products. The purpose of this study is to investigate whether polymorphisms in genes involved in folate metabolism affect the plasma concentrations of Hcy and methylmalonic acid (MMA) along with the concentration of serum folate in individuals with DS. Twelve genetic polymorphisms were investigated in 90 individuals with DS (median age 1.29 years, range 0.07-30.35 years; 49 male and 41 female). Genotyping for the polymorphisms was performed either by polymerase chain reaction (PCR) based techniques or by direct sequencing. Plasma concentrations of Hcy and MMA were measured by liquid chromatography-tandem mass spectrometry as previously described, and serum folate was quantified using a competitive immunoassay. Our results indicate that the MTHFR C677T, MTR A2756G, TC2 C776G and BHMT G742A polymorphisms along with MMA concentration are predictors of Hcy concentration. They also show that age and Hcy concentration are predictors of MMA concentration. These findings could help to understand how genetic variation impacts folate metabolism and what metabolic consequences these variants have in individuals with trisomy 21.

Does soy increase blood counts in myelodysplastic syndromes? / Será que a soja aumenta as contagens sanguíneas em síndrome mielodisplásica?

Myelodysplastic syndromes (MDS) are a group of clonal stem cell diseases characterized by ineffective hematopoiesis, bone marrow hyperproliferation, cytopenias in peripheral blood and risk of transformation into acute leukemia. We decided to investigate the effects of a soy concentrate on MDS patients based on the follow-up results of a 61 year-old Japanese female patient who was diagnosed with MDS and refractory cytopenia with multilineage dysplasia in 2003 (hemoglobin = 11g/dL; white blood cells count = 2,500/uL and platelets = 25,000/uL; marrow with mild dysplasia and normal karyotype; paroxysmal nocturnal hemoglobinuria was excluded). She started using soy as a dietary supplementation in May 2004 and presented a gradual increment in blood counts, achieving normalization approximately eight months afterwards. Among the soy components, the main compounds with anti-carcinogenic activity are the isoflavones (genistein and daidzein). Based on these lines of evidence, we proposed to administer daily a standard soy concentrate to 14 MDS out-patients for a minimum period of three months and maximum of 12 months, in an attempt to evaluate prospectively the possible increase in hemoglobin, neutrophils and platelet counts. A historical control group was used to compare results. The use of a soy concentrate in a standardized manner was associated with an increase in neutrophil and/or platelet counts in some cases, but spontaneous increments were also observed in historical controls. This preliminary study does not allow establishing a relation between soy supplementation and blood cell count increase.

As síndromes mielodisplásicas (SMD) são um grupo das doenças clonais de células-tronco caracterizado por hematopoese ineficaz, hiperproliferação de medula óssea, citopenias no sangue periférico e risco de transformação para leucemia aguda. Decidimos investigar os efeitos de um concentrado de soja em pacientes com SMD com base no fato de termos o seguimento de uma paciente japonesa, de 61 anos de idade, que foi diagnosticada em 2003 com SMD, citopenia refratária com displasia subtipo multilinhagens (hemoglobina = 11 g/dL; contagem de glóbulos brancos = 2.500/uL e plaquetas = 25.000/uL; medula com displasia leve e cariótipo normal; hemoglobinúria paroxística excluída), e que começou a usar a soja como suplemento alimentar em maio de 2004, apresentando gradual aumento da contagem das células sanguíneas, atingindo a normalização cerca de oito meses depois. Entre os componentes da soja, os principais compostos com propriedades anticarcinogênese são as isoflavonas (Ge nisteína e daidzeína). Com base nessas linhas de evidência, foi proposto oferecer diariamente um concentrado de soja padrão, por um período mínimo de três meses e máximo de doze meses, a 14 pacientes ambulatoriais, na tentativa de avaliar, prospectivamente, o possível aumento de hemoglobina, neutrófilos e plaquetas. Um grupo controle histórico foi utilizado para comparar os resultados. O uso de um concentrado de soja de forma padronizada foi associado ao aumento na contagem de neutrófilos e/ou de plaquetas em alguns casos, mas aumentos espontâneos também foram observados em controles históricos. Este estudo preliminar não permite estabelecer relação entre o uso de soja e o aumento na contagem sanguínea.

Novel mutations in the SH3BP2 gene associated with sporadic central giant cell lesions and cherubism.

Central giant cell lesion (CGCL) is a reactive bone lesion that occurs mainly in the mandible, characterized by the multinucleated osteoclast-like giant cells in a background of oval to spindle-shaped mononuclear cells. The etiology is unknown and occurs more commonly in young adults. Cherubism, a rare disease found predominantly in females has histologic characteristics indistinguishable from those of CGCL and is caused by mutations mostly present in exon 9 of the SH3BP2 gene. In this study, we investigated four cases of CGCL and one case of cherubism. DNA was extracted from peripheral blood and tumor tissue and all coding and flanking regions of the SH3BP2 amplified by PCR and directly sequenced to identify underlying mutations. Two novel mutations were found; a heterozygous missense mutation c.1442A>T (Q481L) in exon 11 in one sporadic case of CGCL and a heterozygous germline and tumor tissue missense mutation c.320C>T (T107M) in exon 4 in one patient with cherubism. These findings open a new window to investigate the possible relationship between the pathogenesis of the cherubism and CGCL.

Random amplification of polymorphic DNA reveals clonal relationships among enteropathogenic Escherichia coli isolated from non-human primates and humans

Enteropathogenic Escherichia coli (EPEC) strains are important agents of infantile diarrhea all over the world, gaining even greater importance in developing countries. EPEC have also been isolated from various animal species, but most isolates belong to serotypes that differ from those recovered from humans. However, it has been demonstrated that several isolates from non-human primates belong to the serogroups and/or serotypes related to those implicated in human disease. The objective of this study was to evaluate the genetic differences between thirteen strains isolated from non-human primates and the same number of strains isolated from human infections. Human isolates belonged to the same serogroup/serotype as the monkey strains and the evaluation was done by analysis of random amplified polymorphic DNA. Dendrogram analysis showed that there was no clustering between human and monkey strains. Human and non-human isolates of the EPEC serotypes O127:H40 and O128:H2 shared 90 and 87 percent of their bands, respectively, indicating strong genomic similarity between the strains, leading to the speculation that they may have arisen from the same pathogenic clone. To our knowledge, this study is the first one comparing genomic similarity between human and non-human primate strains and the results provide further evidence that monkey EPEC strains correlate with human EPEC, as suggested in a previous investigation.

Anticorpos para Borrelia burgdorferi em indivíduos que trabalham com animais silvestres / Antibodies specific to Borrelia burgdorferi in wild animal workers

The occurrence of serum antibodies to Borrelia burgdorferi in professionals in close contact with wild animals was determined. Seventy eight technicians workers coming from two São Paulo public institutions housing wild animals had their blood collected (serum samples). All samples were submitted to ELISA for IgM and IgG antibodies against Borrelia burgdorferi. The results showed five positive (6.4 percent), two suspect (2.6 percent) and 71 negatives (91 percent) samples. Based on positive results it is concluded that the infection level is higher to that detected in the general population and similar to values of endemic areas, concluding that this assessed population could be considered at risk for Lyme disease.

Bacterial septicemia in water snakes (Helicops modestus) in Brazil

Relata-se infecção septicêmica em cobras d’água (Helicops modestus) causada por Proteus vulgaris. Os animais iniciavam o processo apresentando abscessos puntiformes de coloração esbranquiçada na pele e após rápida evolução, morriam (100% de mortalidade). Descreve-se, ainda, a sensibilidade do P. vulgaris isolado dos animais a drogas antibacterianas.

Purine DNA adducts of 4,5-dioxovaleric acid and 2,4-decadienal.

The present overview describes recent findings on the formation of cyclic adducts of purine DNA bases after reaction with two aldehyde compounds, 4,5-dioxovaleric acid (DOVA) and 2,4-decadlenal (DDE), which are involved in 5-aminolaevulinic acid (ALA) accumulation and lipid peroxidation, respectively. ALA accumulates under pathological conditions and is associated with an increased incidence of liver cancer. The final oxidation product of ALA, DOVA, is an efficient alkylating agent of the guanine moieties in both nucleoside and isolated DNA. Adducts were produced through the formation of a Schiff base involving the N2-amino group of 2'-deoxyguanosine and the ketone function of DOVA, respectively. DDE is an important breakdown product of lipid peroxidation. It is cytotoxic to mammalian cells and is known to be implicated in DNA damage. It can bind to 2'-deoxyadenosine, yielding highly fluorescent products, including 1,N6-etheno-2'-deoxyadenosine and two other, related adducts. The reaction mechanism for the formation of DDE-2'-deoxyadenosine adducts involves epoxidation of DDE and subsequent addition of the resulting reactive intermediates to the N6 amino group of 2'-deoxyadenosine, followed by cyclization at the N1 site. Formation of endogenous DNA adducts may contribute to the genotoxic potential of ALA and DDE.

Dermatoses observadas no homem e em animais de laboratório, domésticos e silvestres em São Paulo. Levantamento retrospectivo / Dermatoses observed in man, laboratory, domestic and wild animals, in Sao Paulo, Brazil. Restrospective study

Five hundred and forty-five samples of hair, feathers, skin, scale and nails scrapings of 28 different species, including humans domestic animals, laboratory animals and wild animals were analysed. Samples were examined by direct microscopy after 30 per cent KOH clarification, and cultured in dextrose agar Sabouraud, Mycobiotic agar (dextrose 10 g/L, chlormicetine 0.05 g/L, cycloheximide 0.5 g/L), and in brain heart infusion agar (BHI agar). The results obtained after isolation and identification were: bacteria 53.94 per cent, non-dermatophytes mycelial fungi 42.56 per cent, dermatophytes 17.43 per cent, yeasts 13.57 per cent, and acari 2.93 per cent, and 19.26 per cent were negative. Staphylococcus were the most prevalent bacteria and among dermatophytes the most prevalent were M. canis; among non-dermatophytes mycelial the Aspergillus sp, and Candida sp were the most common yeasts isolated

Surtos interespecíficos de dermatomicoses por Microsporum canis e Microsporum gypseum / Human and animal dermatomycosis: outbreaks of Microsporum canis and Microsporum gypseum

As dermatomicoses dos animais domésticos constituem zoonoses importantes, uma vez que estes mantêm estreito contato com a espécie humana, dada a alta infectividade observada nesses processos. Relata-se ocorrência de sete surtos de dermatomicoses, um por M. gypseum envolvendo um gato e um indivíduo do sexo feminino e os outros por M. canis envolvendo 20 indivíduos da espécie humana (adultos, jovens e crianças de ambos os sexos), 5 cäes, 16 gatos e um macaco gibäo (Hylobates lar)

Surtos de salmonelose (Salmonella typhimurium) em Cavia porcellus / Salmonellosis (Salmonella typhimurium) outbreaks in Cavia porcellus

Relatam-se dois surtos de salmonelose em cobaias (Cavia porcellus), envolvendo um total de 70 animais mantidos em biotério, que manifestaram emaciaçäo progressiva, prostraçäo, hipertermia, pelos arrepiados e rarefeitos. Após o início dos sintomas, 48,57 por cento dos animais (34/70) evoluíram para a morte, sendo os demais sacrificados com o intuito de se controlar os surtos. Na necrópsia, colheram-se fragmentos de fígado, baço, rins, pulmöes e intestinos para exames histopatológico e microbiológico. Ao exame microbiológico obteve-se o isolamento de cepas caracterizadas bioquímica e sorológicamente como Salmonella typhimurium. O exame histopatológico dos fragmentos de órgäos corados pela hematoxilina-eosina revelou a presença de lesöes condizentes com quadro septicêmico por este microrganismo. O presenta trabalho alerta quanto ao risco existente na utilizaçäo de animais de experimentaçäo portadores dessa bactéria, já que poderiam comprometer experimentos por uma possível interferência nos resultados, além de representarem fonte de infecçäo em biotérios e laboratórios

Illeo biliar. Presentacion de 18 pacientes. / Biliary ileus. Report of 18 patients

Se presentan 18 casos de ileo biliar estudiados en un periodo de 7 anos en el Hospital General del Centro Medico Nacional.Existio predominio del sexo femenino (94, 4%), con edades de 46 a 85 anos. Todos los pacientes presentaron cuadro de obstruccion intestinal, tanto clinica como radiologicamente. El diagnostico preoperatorio de ileo biliar solo se realizo en el 16.7% pero al efectuar un analisis retrospectivo de los estudios radiologicos se llego a la conclusion que debio ser mas elevado. El tipo de fistula biliar interna mas frecuente fue la colecistoduodenal (66,6%).Diez y siete enfermos fueron tratados quirurgicamente y la mortalidad global fue de 5.6%. La operacion con la que se obtuvieron mejores resultados fue la enterotomia y extraccion del calculo, no se recomienda el cierre de la fistula de primera intencion. En los pacientes con ileo biliar debe tratarse de llegar a un diagnostico correcto lo mas pronto posible, para efectuar el tratamiento oportuno y disminuir la morbimortalidad