Mitochondrial genome analysis in penile carcinoma.

Penile cancer is a rare neoplasm that seems to be linked to socio-economic differences. Mitochondrial genome alterations are common in many tumors types and are reported as regulating oxidative metabolism and impacting tumorigenesis. In this study, we evaluate for the first time the mitochondrial genome in penile carcinoma (PeCa), aiming to evaluate heteroplasmy, mitochondrial DNA (mtDNA) mutational load and mtDNA content in Penile tumors. Using next generation sequencing (NGS), we sequenced the mitochondrial genome of 13 penile tumors and 12 non-neoplastic tissue samples, which allowed us to identify mtDNA variants and heteroplasmy. We further evaluated variant's pathogenicity using Mutpred predictive software and calculated mtDNA content using quantitative PCR. Mitochondrial genome sequencing revealed an increase number of non-synonymous variants in the tumor tissue, along with higher frequency of heteroplasmy and mtDNA depletion in penile tumors, suggesting an increased mitochondrial instability in penile tumors. We also described a list of mitochondrial variants found in penile tumor and normal tissue, including five novel variants found in the tumoral tissue. Our results showed an increased mitochondrial genome instability in penile tumors. We also suggest that mitochondrial DNA copy number (mtDNAcn) and mtDNA variants may act together to imbalance mitochondrial function in PeCa. The better understanding of mitochondrial biology can bring new insights on mechanisms and open a new field for therapy in PeCa.

Levantamento sobre o uso de plantas medicinais com a terapêutica anticâncer por pacientes da Unidade Oncológica de Anápolis / Survey on medicinal plants used with anticancer therapy by patients from Anapolis Oncology Unit

Este estudo teve por objetivo conhecer o perfil dos pacientes em tratamento contra o câncer da Unidade Oncológica de Anápolis quanto ao uso de plantas medicinais. Foram entrevistados 59 pacientes (42,12% da população estudada) por meio de questionários avaliativos enfatizando características sócio-demográficas e dados referentes à utilização de plantas medicinais. Evidenciou-se o uso indiscriminado de plantas medicinais entre os pacientes. A maioria dos entrevistados compartilha a opinião errônea de que plantas medicinais não fazem mal. A orientação sobre a forma de utilização das plantas ocorre, principalmente, pela informação de familiares ou amigos, e os profissionais da saúde muitas vezes são ignorados neste processo. Constatou-se que as plantas medicinais são utilizadas para o tratamento de enfermidades de baixa a alta gravidade, como o câncer. Dentre as 14 espécies usadas pelos pacientes com finalidade antineoplásica as mais mencionadas foram as popularmente conhecidas como noni, babosa, graviola e romã. Apesar de alguns estudos relatarem atividade antineoplásica ou quimiopreventiva para algumas espécies vegetais, muitas delas podem ser tóxicas ou apresentar potencial risco quando usadas concomitantemente ao tratamento convencional. Desta forma, observa-se que é preciso mais profissionais especializados para orientação sobre o risco de reações adversas e interações medicamentosas no que se refere ao uso de espécies vegetais e a terapêutica do câncer.

This study aims to survey the profile ofcancer patients in the Oncology Unit of Anápolis in relation to the use of medicinal plants. To evaluate socio-demographic characteristics and to retrieve data about on the use of medicinal plants, a questionnaire was filled by 59 patients, or 42.12% of the population under study. A widespread use of plants was found. Most of patients share the wrong idea that medicinal plants are not harmful. Guidance on the use of medicinal plants is given primarily by relatives and friends, and health professional are often ignored in this process. Among the 14 species used by patients with antineoplastic purpose, the most mentioned were popularly species known as "noni", "aloe", "soursop" and "pomegranate". Although some studies have reported chemopreventive or antineoplastic activity for some plant species, many may be toxic or show potential risk when used concurrently with conventional treatment. Thus, we can observe that there is a need for more specialized professionals for the guidance on the risk of adverse reactions and drug interactions in relation to the use of plant species and cancer therapeutics.

Vasodilator effect of angiotensin-(1-7) in mature and sponge-induced neovasculature.

Angiotensin-(1-7) (Ang-(1-7)), a peptide constituent of the renin-angiotensin system, has been shown to act as a vasodilator mediator in pre-existing (skin) and newly formed vasculatures (14-day-old sponge implants). Blood flow was determined by the outflow rate of sodium fluorescein applied intradermally or intraimplant and the results were expressed in t(1/2) values (time taken for the fluorescence to reach 50% of the peak in the systemic circulation). We showed that the t(1/2) value was significantly lower (4.1+/-0.46) in the implants compared with the cutaneous vasculature (5.7+/-0.5). Ang-(1-7) 20 ng was able to decrease t(1/2) values in both vasculatures. The specific receptor antagonist, D-Ala7-Ang-(1-7) (A-779), prevented Ang-(1-7)-induced vasodilation and altered the basal vascular tone of the implants. The vasodilator effect was also abolished by nitric oxide (NO) synthase inhibitors in both vasculatures and by indomethacin in the implant. Selective AT(1) and AT(2) receptor antagonists did not alter the vasodilation induced by the peptide. These results establish the vasodilator effect of Ang-(1-7) in the cutaneous and implant vasculature and that the peptide is produced endogenously by the fibrovascular tissue, and suggest that this peptide contributes for the vasodilation found in newly formed vascular beds (wound healing, chronic inflammatory processes and tumors).

Spotted fever in Brazil: a seroepidemiological study and description of clinical cases in an endemic area in the state of São Paulo.

During 1985-1995, illnesses clinically and epidemiologically compatible with Brazilian spotted fever were identified in 17 patients in the county of Pedreira, in the state of São Paulo, Brazil. Spotted-fever group rickettsial infection was confirmed by serology and/or immunostaining of tissues in 10 of these patients. Immunostaining confirmed infection in a 37-year-old pregnant patient, although rickettsial antigens were not demonstrable in the tissues of the fetus. A serosurvey was conducted in four localities in the county to determine the prevalence of subclinical or asymptomatic infections with spotted fever group rickettsiae. Five hundred and twenty-five blood samples were tested by an indirect immunofluorescence assay for antibodies reactive with Rickettsia rickettsii. Twenty-two (4.2%) of these samples demonstrated titers > or = 1:64. The results indicate that Brazilian spotted fever is endemic within this region of Brazil.

Rickettsiae-infected ticks in an endemic area of spotted fever in the State of Minas Gerais, Brazil.

A study on tick-borne rickettsiosis was developed in the county of Santa Cruz do Escalvado, State of Minas Gerais, Brazil, where a clinical case of the disease, confirmed by necropsy, had been reported. Of the 1,254 ticks collected, 1,061 belonged to the Amblyomma genus, 57 to the Rhipicephalus sanguineus species, 81 to Boophilus microplus, and 46 to Anocentor nitens. The hemolymph test associated with Giménez staining showed that 18 of the 221 A. cajennense specimens, 1 of the 16 R. sanguineus, 1 of the 22 B. microplus, 3 of the A. nitens, and 1 of the A. ovale contained rickettsia-like microorganisms. Only 3 A. cajennense ticks were positive under direct immunofluorescence. A. cajennense was the only species found on humans.

Epidemiological aspects of the Brazilian spotted fever: seasonal activity of ticks collected in an endemic area in São Paulo, Brazil

Carrapatos de vegetação e de animais foram coletados mensalmente durante o período de um ano (1993-1994) em uma área endêmica de febre maculosa brasileira no município de Pedreira, São Paulo. Seis espécies de carrapatos foram identificadas Amblyomma cajennense, Amblyomma cooperi, Amblyomma triste, Anocentor nitens, Rhipicephalus sanguineus e Boophilus microplus. Somente a primeira espécie foi suficientemente abundante para permitir um estudo quantitativo com atividade sazonal, embora a distribuição e fonte de captura de outras espécies fossem observadas e aqui relatadas. Estas informações são correlacionadas com a epidemiologia da rickettsiose transmitida por carrapato.

Ticks were collected from vegetation and animals at monthly intervals during one year (1993-1994) in an endemic area of Brazilian spotted fever in the Country of Pedreira, State of São Paulo. Six species of ticks were identified Amblyomma cajennense, Amblyomma cooperi, Amblyomma triste, Anocentor nitens, Rhipicephalus sanguineus and Boophilus microplus. Only the first species was sufficiently numerous to permit a quantitative study with seasonal activity, although the distribution and source of capture of other species were observed and are reported. This information is correlated with the epidemiology of tick-borne rickettsiosis.

Epidemiological aspects of the Brazilian spotted fever: seasonal activity of ticks collected in an endemic area in São Paulo, Brazil.

Ticks were collected from vegetation and animals at monthly intervals during one year (1993-1994) in an endemic area of Brazilian spotted fever in the Country of Pedreira, State of São Paulo. Six species of ticks were identified Amblyomma cajennense, Amblyomma cooperi, Amblyomma triste, Anocentor nitens, Rhipicephalus sanguineus and Boophilus microplus. Only the first species was sufficiently numerous to permit a quantitative study with seasonal activity, although the distribution and source of capture of other species were observed and are reported. This information is correlated with the epidemiology of tick-borne rickettsiosis.

Sponge-induced angiogenesis in mice and the pharmacological reactivity of the neovasculature quantitated by a fluorimetric method.

Sponge-induced angiogenesis in mice and pharmacological reactivity of the neovasculature have been determined by a fluorimetric method. Pharmacokinetic studies following subcutaneous, intradermal, and intraimplant administration of sodium fluorescein resulted in a biphasic curve from which estimation of t1/2 for absorption and elimination of the dye were possible. Following topical injection of the dye at days 1, 4, 7, 10, and 14 postimplantation, measurement of fluorchrome generated emission in the systemic circulation reflected the development of blood flow in and around the implants and the interaction of the angiogenic site with the systemic circulation. The t1/2 values for the fluorescence peak in the bloodstream decreased steadily from an initial value of 6.41 +/- 0.28 min (avascular implant) to 2.78 +/- 0.23 min in fully vascularized implants (day 14). The reactivity of the neovasculature to ET-1 was dose-dependent and similar to the skin vasculature. By contrast, no reactivity to histamine was detected in the implant blood vessels, whereas it was present in the skin. These results show that the pharmacological response of the neovasculature differs from the response of mature blood vessels. The angiogenic stimulus (bFGF, 300 ng daily) decreased t1/2 for the fluorescence peak, whereas dexamethasone (1 mg/kg) increased it. Parallel histological studies corroborated the functional findings. These observations indicate the suitability of this assay to study angiogenesis, functional and pharmacological characterization of the neovasculature, and the interaction of the angiogenic site with the systemic circulation.

Angiotensin-II-induced angiogenesis in sponge implants in mice.

Stimulators of angiogenesis hold potential in promoting the development of collateral circulation in ischaemic tissue and accelerating would healing, but promote pathological vasoformation in angiogenesis-dependent diseases (solid tumours, atherosclerosis). The renin-angiotensin system is implicated in both beneficial angiogenesis and pathological vascular growth. We investigated the angiogenic activity of angiotensin II (AII) in a sponge implant model in mice; this peptide enhanced angiogenesis, as well as glycosaminoglycan (GAG, chondroitin sulfate proteoglycan) and protein synthesis in sponge matrix in mice in a dose-dependent fashion. Extensive angiogenesis was achieved with AII (1 microgram), which gave no significant increase in wet weight and protein and only a small effect on GAG. In the implants treated with AII (2 micrograms) no further increase in angiogenesis was observed, whereas a marked effect was shown in wet weight (326 +/- 15 vs. 424 +/- 27 mg), total protein (18 +/- 1 vs. 25 +/- 1 micrograms/ww) and GAG (98 +/- 10 vs. 160 +/- 13 ng/ww). The local blood flow has been determined by measuring the washout rate of 133Xe injected into the implants, correlated with histological evidence of vessel growth. This model of angiogenesis has allowed sequential studies of fibrovascular tissue infiltration simultaneously with histological and biochemical parameters of angiogenesis.

Epidemiological aspects of the Brazilian spotted fever: serological survey of dogs and horses in an endemic area in the State of São Paulo, Brazil.

In order to obtain information on Brazilian spotted fever, a study in domestic animals was performed in the County of Pedreira, State of São Paulo, Brazil, where 17 human cases had been notified. Serum samples obtained from animals were tested by indirect immunofluorescence for detectable antibodies to spotted fever-group rickettsiae. Seropositivity was revealed in 12 (36.4%) of 33 dogs and seven (77.8%) of nine horses from the endemic area. For comparison, blood samples from dogs and horses from non endemic area were tested and four (12.9%) of 31 dogs and three (27.3%) of 11 horses were positive. The highest titers of antibodies by IFA (IgG > or = 1:1024) were found only in three dogs and six horses from endemic area. The results suggest that dogs as horses may serve as environmental sentinels for establishing the prevalence of foci of spotted fever in Brazil.

Primary isolation of spotted fever group rickettsiae from Amblyomma cooperi collected from Hydrochaeris hydrochaeris in Brazil.

This paper reports the first isolation of a spotted fever group rickettsia from an Amblyomma cooperi ixodid collected from a capybara (Hydrochaeris hydrochaeris) in an endemic area of spotted fever in the County of Pedreira, State of São Paulo, Brazil. Isolation was performed in Vero cell culture and submitted to immunofluorescence, using antibody from Rickettsia rickettsii-positive human serum.

Rocky Mountain spotted fever in an endemic area in Minas Gerais, Brazil.

Only one species of spotted fever-group rickettsiae that is pathogenic for humans has been isolated in Brazil, where few physicians are familiar with this disease. In order to obtain information on tick-borne rickettsiosis, a study was performed in the County of Santa Cruz do Escalvado, State of Minas Gerais, Brazil, where a fatal clinical case confirmed by specific immunofluorescence had been reported. Serum samples obtained from 679 humans and 96 dogs were tested by indirect immunofluorescence for detectable antibodies to spotted fever-group rickettsiae, the criterion for a positive result being a titer > or = 1:64. Seropositivity was detected in 7.14% of the humans sera examined and 13.68% of the dogs. We discuss the significance of these findings and formulate some questions, emphasizing the need for further investigation.

Comparative evaluation of a simple and sensitive assay for detection of orthomyxo and paramyxoviruses

Studies were done to evaluate comparatively the traditional HA assay and a more recently introduced lectin-neuraminidase (LN) methodologyin search of a simple and sensitive assay for virus detection during laboratorial diagnosis. The results proved the value of LN assay as a sensitive methodologyfor detection of virus particles, presenting results at least equal to those obtained by HA (hemagglutination) assay, with significant values of accumulated frequencies for LN/HA factors (ratios between LN and HA titers) higher than two. The accumulated values of frequencies for LN/HA factors as high as four were very significant, 72.7 (per cent) for influenzavirus and 60.7 (per cent) for Newcastle disease virus (NDV), moreover accumulated frequencies for LN/HA factors even as high as 32 were due to influenzavirus (45.4 per cent) and NDV (7.2 per cent) samples. After the storage period, most of those concentraded samples that even did not present HA titers could be detected through LN assay, demonstrating a lower threshold for virus detection

Comparative evaluation of a simple and sensitive assay for detection of orthomyxo and paramyxoviruses.

Studies were done to evaluate comparatively the traditional HA assay and a more recently introduced lectin-neuraminidase (LN) methodology in search of a simple and sensitive assay for virus detection during laboratorial diagnosis. The results proved the value of LN assay as a sensitive methodology for detection of virus particles, presenting results at least equal to those obtained by HA (hemagglutination) assay, with significant values of accumulated frequencies for LN/HA factors (ratios between LN and HA titers) higher than two. The accumulated values of frequencies for LN/HA factors as high as four were very significant, 72.7% for influenzavirus and 60.7% for Newcastle disease virus (NDV), moreover accumulated frequencies for LN/HA factors even as high as 32 were due to influenzavirus (45.4%) and NDV (7.2%) samples. After the storage period, most of those concentraded samples that even did not present HA titers could be detected through LN assay, demonstrating a lower threshold for virus detection.

Caracterizaçao e perfil de sensibilidade de 45 amostras de Staphylococcus coagulase negativa de origem hospitalar / Sensitivity and characteristics of 45 strains coagulase-negative Staphylococcus sp isolated from clinical specimens

No presente trabalho foram analisadas 45 amostras de Staphylococcus coagulase negativa isoladas de especimes clinicos, quanto a diversas caracteristicas e propriedades biologicas. Alem da classificaçao em especies, foram efetuados estudos sobre a sensibilidade a antimicrobianos e determinada a concentraçao minima inibitoria destas amostras frente a vancomicina e a oxacilina. Cinco especies foram identificadas pelo metodo convencional (Kloos & Schleifer, 1975) associado a alguns testes do Micrometodo Bactrey, e corresponderam: S.saprohyticus (6); S. epidermidis (20); S. hominis (4); S. haemolyticus (7); S. warnery (5). Tres amostras nao foram identificadas. Uma percentagem elevada (84,4 por cento ) de amostras apresentaram resistencia a penicilina, em menor grau, a canamicina (55,6 por cento ), gentamicina (48,9 por cento ), fosfomicina (44,4 por cento ) e, oxacilina e sulfametoxazol-trimetoprim (37,8 por cento ). Foram encontradas amostras (66,6 por cento ) multi-resistentes. Todas as amostras foram testadas quanto a produçao de beta-lactamase, 35 tiveram resultado positivo, das quais 20 eram S. epidermidis. Na determinaçao da concentraçao minima inibitoria a oxacilina apresentou uma percentagem de amostras resistentes (72,1 por cento ) bem maior que no teste de sensibilidade a antimicrobianos por difusao em agar frente a este antibiotico (39,5 por cento )