Methylation of p16 ink4a promoter is independent of human papillomavirus DNA physical state: a comparison between cervical pre-neoplastic and neoplastic samples

BACKGROUND Epigenetic modifications in host cells, like p16 ink4a methylation, have been considered as putative complementary mechanisms for cancer development. Because only a small proportion of infected women develop cervical cancer, other factors might be involved in carcinogenesis, either independently or in association with high-risk human papillomavirus (HR-HPV) infections, including epigenetic factors. OBJECTIVES We hypothesised that p16 ink4a methylation might have a role in cancer development driven by HPV16, mainly in the presence of intact E1/E2 genes. Thus, our objectives were to assess the status of p16 ink4a methylation and the HPV16 E1/E2 integrity in samples in different stages of cervical diseases. METHODS Presence of HPV16 was determined by E6 type-specific polymerase chain reaction (PCR). Methylation status of the p16 ink4a promoter was assessed by methylation-specific PCR in 87 cervical specimens comprising 29 low-grade (LSIL), 41 high-grade (HSIL) lesions, and 17 cervical cancers (CC). Characterisation of E1 and E2 disruption (as an indirect indicator of the presence of episomal viral DNA) was performed by PCR amplifications. FINDINGS We observed a significantly increased trend (nptrend = 0.0320) in the proportion of methylated p16 ink4a in cervical samples during cancer development. Concomitant E1 and E2 disruptions were the most frequent pattern found in all groups: CC (76%), HSIL (54%), and LSIL (73%). No statistically significant differences between p16 ink4a methylation and E1/E2 integrity, in histological groups, was observed. MAIN CONCLUSIONS There was an increase in methylation of the p16 ink4a promoter from pre-neoplastic lesions to cancer. Additionally, a high frequency of E1/E2 disruptions in LSIL/HSIL suggested that viral DNA integration was an early event in cervical disease. Moreover, the methylation status was apparently independent of HPV16 integrity.

Doenças negligenciadas no município de sabará: casos, portadores e percepções / Neglected diseases in the city of sabará (mg): cases, affected people and perceptions

As doenças negligenciadas (DNs) são causadas por agentes infecciosos e parasitários, e na maioria das vezes acometem populações de baixa renda. A atenção primária à saúde (APS) atende, aproximadamente, 85% dos problemas de saúde da população, incluindo parte dos serviços relacionados aos portadores das DNs. Sabará, Minas Gerais, vem registrando um aumento no número de notificações para várias DNs transmitidas por vetores e/ou com hospedeiros intermediários envolvidos em seu ciclo. Nessa perspectiva, o presente trabalho teve como objetivo levantar o número de notificações de DNs transmitidas por vetores e/ou com hospedeiros intermediários envolvidos em seu ciclo entre os anos 2008 a 2014 no município, analisar o perfil sociodemográfico dos portadores e avaliar a percepção de profissionais da Estratégia de Saúde da Família (ESF) sobre aspectos relacionados a essas doenças. Para o levantamento das DNs e do perfil sociodemográfico de seus portadores foram analisadas as fichas de notificação compulsória das DNs. As DNs transmitidas por vetores e/ou com hospedeiros intermediários envolvidos em seu ciclo com maior número de notificações em Sabará foram: dengue, esquistossomose, leishmaniose visceral e leishmaniose tegumentar americana (LTA). Em relação à idade, a leishmaniose visceral acometeu, fundamentalmente, crianças, e a dengue, a LTA e a esquistossomose, adultos. Para a avaliação da percepção dos profissionais da ESF sobre as DNs foram aplicados questionários estruturados aos enfermeiros, Agentes Comunitários de Saúde (ACSs) e auxiliares/técnicos de enfermagem de 09 programas de saúde. Através desse instrumento pode-se evidenciar um desconhecimento sobre as principais endemias, vetores e hospedeiros intermediários, ausência de conhecimento sobre o fluxo de atendimento das DNs, insuficiente articulação entre Atenção Primária a Saúde e demais níveis de atenção, e insipiência de informações disponibilizadas pela APS à população. Reforça-se que Sabará é um município endêmico para várias DNs transmitidas por vetores e/ou com hospedeiros intermediários envolvidos em seu ciclo, e que os profissionais da APS do município desconhecem essa realidade.

Neglected diseases (NDs) are caused by infectious and parasitic agents, and most often affect low-income populations. Primary health care (PHC) covers approximately 85% of the population's health issues, including part of the services related to those affected with NDs. The city of Sabará in Minas Gerais (Brazil) has registered an increase in the number of notifications for various NDs transmitted by vectors and/or intermediate hosts involved in its cycle. In this perspective, this study aims at raising the number of notifications regarding NDs transmitted by vectors and/or intermediate hosts between the years 2008-2014 in the city by analyzing the socio-demographic profile of patients and evaluating the perception of professionals in the Family Health Strategy (FHS) on aspects related to these diseases. In order to collect data related to the NDs and patient socio-demographics, ND compulsory notification records were analyzed. The NDs transmitted by vectors and/or intermediate hosts with the highest number of notifications in Sabará were dengue fever, schistosomiasis, visceral leishmaniasis and American cutaneous leishmaniasis. Regarding age, visceral leishmaniasis occurred mainly in children, whereas dengue fever, American cutaneous leishmaniasis and schistosomiasis occurred in adults. In order to evaluate the perception of the FHS professionals on NDs, structured questionnaires were applied to nurses, community health workers (CHWs) and nursing assistants/technicians of nine health programs. Through this tool, it was possible to show the lack of knowledge about the major endemic diseases and vectors, lack of knowledge about the flow of care of NDs, insufficient coordination between Primary Health Care and other levels of attention, and lack of information provided from PHC to the population. It must be emphasized that the city of Sabará is an endemic municipality for several NDs transmitted by vectors and/or intermediate hosts, and PHC professionals are not aware of that reality.

Paratuberculose em ruminantes no Brasil / Paratuberculosis in ruminants in Brasil: a review

A paratuberculose ou doença de Johne é uma enterite granulomatosa causada por Mycobacterium avium subsp. paratuberculosis (Map) e comumente afeta ruminantes domésticos, no entanto, pode infectar várias espécies de mamíferos. Está presente nos cinco continentes e é considerada endêmica em algumas regiões pela Organização Internacional de Epizootias (OIE). Pertence à lista de enfermidades notificáveis, que compreende as doenças transmissíveis de importância sócio-econômica e/ou em saúde-pública, cujo controle é necessário para o comércio internacional de animais e alimentos de origem animal. A importância da doença de Johne não se restringe somente aos prejuízos econômicos causados à indústria animal, mas também na possível participação do Map na íleocolite granulomatosa que afeta seres humanos, conhecida como doença de Crohn. No Brasil, a paratuberculose já foi descrita em diversas espécies de ruminantes e em vários estados. Embora os relatos naturais da enfermidade sejam pontuais, acredita-se na possibilidade da transmissão interespecífica e na disseminação do agente através da compra e venda de animais infectados. O objetivo deste artigo foi reunir as informações disponíveis referentes aos aspectos epidemiológicos, clínico-patológicos e laboratoriais da paratuberculose em bovinos, bubalinos, caprinos e ovinos no Brasil, e salientar a necessidade de implementação de medidas de controle sanitário da enfermidade no país, o que possibilitaria a melhoria da qualidade e valorização dos produtos de origem animal no mercado internacional.

Paratuberculosis also known as Johne's disease, is a granulomatous enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP), an acid-fast bacillus that preferentially resides within host intestinal macrophages. The condition is most commonly seen in domestic ruminants, however MAP can also infect other mammalian species. Paratuberculosis shows a global distribution and is considered endemic in some regions. The World Organization for Animal Health (OIE, Office International des Epizooties), have classified paratuberculosis as a notificable disease; considered to be of socio-economic and/or public-health importance, the control of which is necessary for the international trade of animal and animal products. The importance of paratuberculosis is related primarily to economic losses in the animal industry and also because of a potential role for this bacterium in the pathogenesis of Crohn´s disease, a debilitating condition affecting the digestive tract of humans. In Brazil, paratuberculosis has been reported in a variety of ruminant species and shows a broad geographic distribution. The reported incidence of natural cases in Brazil has been limited, but it is believed that interespecific transmission of MAP and dissemination of the agent is driven by the commercialization of infected animals. The main objective of this paper was to collate the published epidemiological, clinic-pathological and diagnostic information in relation to paratuberculosis in cattle, buffaloes, goats and sheep in Brazil. Moreover, it served as a platform to emphasize the requirement to implement sanitary policies for control of MAP in the county, which may serve to improve the quality and value of animal products on international markets.

characteristics women with cervical cancer referred for radiotherapy and /or chemotherapy

Cervical cancer is the most common gynecologic cancer in women in the world, most arise from infection with human papiloma virus. Other host factors also affect the neoplastic progression following initial infection. To determine characteristics of Iraqi women with carcinoma of the uterine cervix during the last 11 years in Iraq. Retrospective study; done on records in Radiotherapy and Nuclear Medicine Hospital- Baghdad; 488 cases of cervical carcinoma referred for chemotherapy and or radiotherapy, from 1999 to 2009. Clinical and pathological data were reviewed and analyzed. The data that was obtained from the radiotherapy and nuclear medicine hospital showed that the women with cervical carcinoma constitute only 2.1% of total women with malignancies during 1999-2009. Highest; were in 2003 and 2004. Largest proportion presented in late stage [62.32%] and only 37.67% with early stage. Squamous cell carcinoma counted 98.38% and only 1.62% were adenocarcinoma. Median and mean age was 45-50. Median parity between 4 and 6. Majority of patients from Baghdad [43.02%], Basra [10.93%] and lowest percentage from North; been collectively [10.46%]. Only 6.55% had positive family history. [53.72%] with history of smoking; significantly associated with late stage. Most common clinical presentation was vaginal bleeding [65.81%]. The most common stage at time of presentation was stage two [36.51%]. Most of referred patients had surgical interventions as total abdominal hysterectomy with bilateral salpingooophorectomy [sub optimal surgery] 60.93%. Most patients with carcinoma of cervix in Iraq presented in late stage due to absence of screening program.

Examination of Geographical, Clinical and Intrahost Variations in the 3' Repeat Region of CagA Gene in Helicobacter pylori

The size variation of the cytoxin-associated protein (cagA), which is dependent on the 3' repeat region (3'RR) of the cagA gene, is known to play a crucial role in the pathogenesis of Helicobacter pylori infection. The present study evaluated the relationship between the 3'RR variation and the geographic distribution, clinical manifestations, and locations of colonization in the stomach. We evaluated the 3'RR of H. pylori isolates from 78 patients with gastric cancer, peptic ulcer, and non-ulcer dyspepsia from Japan, Hong Kong, India, and the United States and assessed the variations of 3'RR according to the geographical and clinical characteristics. Sixty eight (87.2%) patients had the same 650 bp band without geographical differences. The frequency of polymorphisms in the 3'RR did not differ when compared to the clinical manifestations (P=0.868). The length of 3'RR did not differ by location of colonization. In conclusion, the 3'RR variation of cagA gene is not associated with the geographical and clinical characteristics of the patients studied.

Aspectos biológicos de Cirrospilus neotropicus Diez & Fidalgo (Hymenoptera: Eulophidae), parasitóide de Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae) / Biological aspects of Cirrospilus neotropicus Diez & Fidalgo (Hymenoptera: Eulophidae), parasitoid of Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae)

A biologia de Cirrospilus neotropicus Diez & Fidalgo, tendo como hospedeiro lagartas de terceiro ínstar de Phyllocnistis citrella Stainton criadas em folhas de limoeiro æCravoÆ (Citrus limonia Osbeck), foi avaliada em estufa climatizada (25 ± 1ºC; fotofase de 12h), registrando-se a duração e a sobrevivência das fases imaturas, a longevidade, a razão sexual, as taxas de oviposição e a atividade de predação. Dois grupos de fêmeas foram acompanhados, um proveniente de câmaras pupais de P. citrella coletadas em pomares de citros, e, outro de uma criação de laboratório. As fêmeas foram mantidas em caixas gerbox, recebendo, a cada 48h, 12 lagartas de P. citrella, mel e pólen. A duração média do período de ovo a adulto para os indivíduos gerados por fêmeas de campo e de laboratório foi, respectivamente, de 11,8 e 11,6 dias para fêmeas e 11,6 e 10,9 dias para machos. A menor sobrevivência foi registrada para a fase larval na prole de ambos os grupos de fêmeas, 57,3 por cento - campo e 57,4 por cento - laboratório. A longevidade média de fêmeas do campo acasaladas (21,8 dias) foi significativamente superior a das de laboratório acasaladas (16,8 dias). A taxa média de oviposição para as fêmeas de campo (2,8 ovos/dia) foi significativamente superior às das de laboratório (1,6 ovos/dia). O mesmo foi observado para o número médio diário de picadas alimentícias (1,7 lagarta/dia para fêmeas de campo e 0,87 lagarta/dia para as de laboratório). Os dados indicam que C. neotropicus tem potencial como agente de controle biológico de P. citrella.

The biology of Cirrospilus neotropicus Diez & Fidalgo reared on third instar Phyllocnistis citrella Stainton larvae having Citrus limonia Osbeck as host plant, was evaluated under controlled conditions (25 ± 1ºC; 12h photophase). The survival, immatures development, longevity, sex ratio, host feeding and oviposition ratio of C. neotropicus, were registered. Two groups of parasitoid females were evaluated: one, with parasitoids obtained from P. citrella pupae collected in citrus orchards and the other group came from laboratory rearing. These females, after mating, were maintained individually on gerbox containers with honey and pollen as food source. At each 48h, 12 P. citrella larvae were changed. The average biological cycle of the female progenies from orchard and laboratory generation groups were 11.8 and 11.6 days, respectively. The males progenies had biological cycles of 11.6 and 10.9 days, as well. The shortest immature survival period, for both studied groups was the larval (57.3 percent - orchard group and 57.4 percent - lab group). The mated females average longevity (21.8 days) was superior than the non mated ones (9.1 days) for the orchard group. The average daily oviposition rate for this last group was also significantly superior (2.8 eggs /day) than the lab group (1.6 eggs / day). The same trend was observed for the average daily host feeding rate (1.7 larvae /day for the orchard group and 0.9 larvae/ day for the lab group). These data suggest that C. neotropicus have potencial as a biological control agent of P. citrella.

A genomic-scale search for regulatory binding sites in the integration host factor regulon of Escherichia coli K12

We examined general aspects of the DNA-protein interaction between the integration host factor (IHF) global regulator and its regulatory binding sites in the Escherichia coli K12 genome. Two models were developed with distinct weight matrices for the regulatory binding sites recognized by IHF. Using these matrices we performed a genome scale scan and built a set of computationally predicted binding sites for each of the models. The sites found by the model associated with repetitive sequences had a higher score in the sequence to matrix alignment. They were also more rare than the other sites. The sites not associated with repeats rapidly tended to become undistinguishable from the background as statistical stringency was relaxed. We compared our results to the known sites documented in RegulonDB and found new members of the IHF Regulon. The two models exhibit clearly distinct affinity patterns (scores in the sequence to matrix alignments and in the number of regulatory sites), as we vary the stringency of the statistical confidence parameters. We suggest that these differences may play an important role in the dynamics of the network. We concluded that IHF may regulate two genes encoding ATP-dependent RNA helicases. This interaction is not described in RegulonDB, even as a computational prediction. IHF may also regulate RNA modification processes.

Influência do hospedeiro no ciclo e comprovação do ciclo heteroxeno de Anocentor nitens (Neumann, 1897) / Host influency on life cycle and comprovation of the heteroxenous cycle of Anocentor nitens (Neumann, 1897)

Steers and rabbits were used as experimental hosts for Anocenter nitens during scheduled infections, when ticks were recovered from naturally infected horses. Thus, the influence of the host species on the parasite was evaluated mainly considering parameters related to the engorgement period and pre molting of evolutive stages. It was also observed that almost 5% if the metanymphae and a few more than 1% of metalarvae fall off from the host to molt in the evironment, characterizing cycles in one or more hosts. The heteroxenous cycle is reported for the first time occuring in A. nitens.