Implications of intrauterine protein malnutrition on prostate growth, maturation and aging

Aims Maternal malnutrition by low protein diet is associated with an increased incidence of metabolic disorders and decreased male fertility in adult life. This study aimed to assess the impact of maternal protein malnutrition (MPM) on prostate growth, tissue organization and lesion incidence with aging. Main methods Wistar rat dams were distributed into two groups, which were control (NP; fed a normal diet containing 17% protein) or a restricted protein diet (RP, fed a diet containing 6% protein) during gestation. After delivery all mothers and offspring received a normal diet. Biometrical parameters, hormonal levels and prostates were harvested at post-natal days (PND) 30, 120 and 360. Key findings MPM promoted low birth weight, decreased ano-genital distance (AGD) and reduced androgen plasma levels of male pups. Prostatic lobes from RP groups presented reduced glandular weight, epithelial cell height and alveolar diameter. The epithelial cell proliferation and collagen deposition were increased in RP group. Incidences of epithelial dysplasia and prostatitis were higher in the RP offspring than in the NP offspring at PND360. Significance Our findings show that MPM delays prostate development, growth and maturation until adulthood, probably as a result of low testosterone stimuli. The higher incidence of cellular dysplasia and prostatitis suggests that MPM increases prostate susceptibility to diseases with aging. © 2013 Elsevier Inc.

Avaliação do desenvolvimento do sistema genital e da funçao reprodutiva da prole feminina de ratos Wistar expostos à cisplatina durante a peri-puberdade

Pós-graduação em Biologia Geral e Aplicada - IBB

Avaliação da exposição ao propionato de testosterona em ratas fêmeas in utero e lactação

It is known that exposure to substances in the environment can contribute to various reproductive disorders, especially if such exposure occurs during critical periods of development such as the intra-uterine and postnatal. The female reproductive system may be the target of androgens, both as a result of exposure to environmental chemicals, or by pathological conditions (polycystic ovary syndrome or congenital adrenal hyperplasia).Usually, little attention is given off in relation to the study of androgenic effects in the female reproductive axis. This study aims to evaluate the effects of exposure to androgens on the development, structure and reproductive function in rats whose mothers were exposed to testosterone propionate from gestational day 12 (DG12) after weaning - postnatal day 21 (DPN21) . For this purpose, pregnancy rats were divided into four groups: a control group that received corn oil (vehicle) and three groups receiving testosterone propionate in doses of 0.05 mg / kg / day, 0.1 mg / kg / day and 0.2 mg / kg / day, all under the same experimental conditions. The possible effects of exposure were assessed using reproductive parameters, such as a measure of anogenital distance, count areolas / nipples, age at vaginal opening and first estrus (puberty indicative installation), weight and histological evaluation of the reproductive organs ( uterus and ovaries), weight of the kidneys, liver and pituitary hormone levels, regularity of the estrous cycle, sexual behavior and fertility. Such analysis is important in understanding the effects of androgen exposure on the female genital system, especially on the reproductive potential, and processes that may involve morphofunctional changes. In these experimental conditions, it is concluded that treatment with PT caused reduction in body weight and initial masculinization in females without cubs, however, commit further sexual development

The relationship between angiogenesis and the immune response in carcinogenesis and the progression of malignant disease

Recent studies have demonstrated that angiogenesis and suppressed cell- mediated immunity (CMI) play a central role in the pathogenesis of malignant disease facilitating tumour growth, invasion and metastasis. In the majority of tumours, the malignant process is preceded by a pathological condition or exposure to an irritant which itself is associated with the induction of angiogenesis and/or suppressed CMI. These include: cigarette smoking, chronic bronchitis and lung cancer; chronic oesophagitis and oesophageal cancer; chronic viral infections such as human papilloma virus and ano-genital cancers, chronic hepatitis B and C and hepatocellular carcinoma, and Epstein- Barr virus (EBV) and lymphomas; chronic inflammatory conditions such as Crohn's disease and ulcerative colitis and colorectal cancer; asbestos exposure and mesothelioma and excessive sunlight exposure/sunburn and malignant melanoma. Chronic exposure to growth factors (insulin-like growth factor-I in acromegaly), mutations in tumour suppressor genes (TP53 in Li Fraumeni syndrome) and long-term exposure to immunosuppressive agents (cyclosporin A) may also give rise to similar environments and are associated with the development of a range of solid tumours. The increased blood supply would facilitate the development and proliferation of an abnormal clone or clones of cells arising as the result of: (a) an inherited genetic abnormality; and/or (b) acquired somatic mutations, the latter due to local production and/or enhanced delivery of carcinogens and mutagenic growth factors. With progressive detrimental mutations and growth-induced tumour hypoxia, the transformed cell, to a lesser or greater extent, may amplify the angiogenic process and CMI suppression, thereby facilitating further tumour growth and metastasis. There is accumulating evidence that long-term treatment with cyclo-oxygenase inhibitors (aspirin and indomethacin), cytokines such as interferon-α, anti-oestrogens (tamoxifen and raloxifene) and captopril significantly reduces the incidence of solid tumours such as breast and colorectal cancer. These agents are anti-angiogenic and, in the case of aspirin, indomethacin and interferon-α have proven immunomodulatory effects. Collectively these observations indicate that angiogenesis and suppressed CMI play a central role in the development and progression of malignant disease. (C) 2000 Elsevier Science Ltd.

Cancro oral

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Medicina Dentária

Papilomatose oral : considerações a propósito de um caso clínico

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Prophylactic HPV vaccines: Underlying mechanisms

Human papillomavirus virus-like particles (HPV VLP) can be generated by the synthesis and self-assembly in vitro of the major virus capsid protein L1. HPV L1 VLPs are morphologically and antigenically almost identical to native virions, and this technology has been exploited to produce HPV L1 VLP subunit vaccines. The vaccines elicit high titres of anti-L I VLP antibodies that persist at levels 10 times that of natural infections for at least 48 months. At present the assumption is that the protection achieved by these vaccines against incident HPV infection and HPV-associated ano-genital pathology is mediated via serum neutralising Immunoglobulin G (IgG). However, since there have been very few vaccine failures thus far, immune correlates of protection have not been established. The available evidence is that the immunodominant neutralising antibodies generated by L1 VLPs are type-specific and are not cross-neutralising, although highly homologous HPV pairs share minor cross-neutralisation epitopes. Important issues remaining to be addressed include the duration of protection and genotype replacement. (c) 2006 Elsevier Ltd. All rights reserved.

A comparison of the estrous behavior of Holstein-Friesian cows when cubicle-housed and at pasture

This study compared estrous behavior of dairy cows kept in cubicle housing and fed a total mixed ration diet (HOUSED treatment) with that of cows kept at pasture with concentrate supplementation (PASTURE treatment). Behavior was compared both in the 48 h around standing estrus and during the standing estrus period. The 23 spring-calving Holstein-Friesians in each treatment were observed directly three times per day for nine weeks. The occurrence of nine selected behaviors associated with estrus was recorded during 20 min observation sessions. Twelve standing estrus events from each treatment were selected for analysis of the frequency of these nine behaviours over the 48 h around standing estrus. Milk progesterone profiles were used to confirm the dates of standing estrus events. Attempting to mount other cows, sniffing the anogenital region of other cows, resting the chin on other cows, receiving chin rests and head-to-head butts all showed significant changes in frequency in the 48 h around standing estrus in both treatments, reaching a peak during standing estrus (P ≤ 0.05). Mounting other cows increased significantly in the PASTURE treatment around standing estrus (P <0.001), but not in the HOUSED treatment. The frequency of ano-genital sniffs received by the animals in the PASTURE treatment also increased significantly around standing estrus (P <0.01) but not in the HOUSED treatment. When the animals were in standing estrus there was a significantly higher frequency of standing to be mounted in PASTURE than in HOUSED cows (median (q1, q3) PASTURE = 2.5 (1.0, 3.0), HOUSED = 0.0 (0.0, 1.0)) (P <0.01), but no difference in the frequency of the other eight sexual behaviors recorded. HOUSED cows did not exhibit the same increase in mounting during the standing estrus period as PASTURE cows and received fewer mounts in observation sessions during standing estrus. These results have implications for the use of estrus detection systems that rely solely on mounting behavior in cubicle-housed dairy cows. © 2012 Elsevier Inc.

Redução do número de espermatozóides por fêmea suína inseminada por ano.

Na inseminação artificial de suínos, são utilizados de 9 a 12 bilhões de espermatozóides por fêmea inseminada. A utilização de menor número de espermatozóides, sem interferir na performance reprodutiva das fêmeas, poderia otimizar o uso dos machos e reduzir os custos de inseminação. Esta tese foi dividida em três experimentos para avaliar o efeito da redução do número de espermatozóides por fêmea inseminada com a técnica tradicional ou intra-uterina. No experimento 1 foram utilizadas 218 leitoas Camborough 22 inseminadas em três intervalos antes da ovulação (0-12, 13-23 e 24-30 h), com doses inseminantes contendo 1,5 bilhão de espermatozóides e armazenadas por 0-48 h ou 96-120 h. As leitoas receberam uma única inseminação. As fêmeas foram abatidas aos 30,83,7 dias de gestação e o trato genital foi removido para contagem do número de corpos lúteos e embriões totais. A taxa de prenhez foi influenciada pelo intervalo inseminação-ovulação quando o sêmen foi armazenado por 96-120 h (P<0,05). Da mesma forma, a interação tempo de armazenamento do sêmen e o intervalo inseminação-ovulação afetou (P<0,05) o número de embriões totais e a sobrevivência embrionária. Quando o período de armazenamento do sêmen foi de 120 h e o intervalo inseminação-ovulação de 24-30 h, foi observada redução no número de embriões totais e na sobrevivência embrionária. No experimento 2 foi avaliado o efeito do intervalo inseminação-ovulação e do número de espermatozóides na dose inseminante em fêmeas submetidas à inseminação intra-uterina. Foram utilizadas 66 matrizes pluríparas da linhagem Camborough 22. As fêmeas foram distribuídas em quatro tratamentos (doses de 1 ou 2 bilhões de espermatozóides diluídos em 60 ml e intervalo inseminação-ovulação de 0-24 e 25-36 h). As fêmeas receberam uma única inseminação intra-uterina. No momento da inseminação, não foi observado refluxo de sêmen. A passagem do cateter pela cérvix foi possível em todas as fêmeas. Foi observada presença de sangue em 1,7% das fêmeas. Aos 314,3 dias de gestação, as fêmeas foram abatidas e o trato genital foi removido para a contagem dos corpos lúteos e número de embriões totais. A taxa de prenhez e a sobrevivência embrionária não foram afetadas pelo número de espermatozóides ou pelo intervalo inseminação-ovulação. O número de embriões totais não foi influenciado pelo número de espermatozóides, mas foi reduzido para o intervalo inseminação-ovulação de 25-36 h comparado a 0-24 h (P<0,05). No experimento 3 foram utilizadas 298 fêmeas Camborough 22 com o objetivo de comparar o desempenho reprodutivo de fêmeas submetidas à inseminação intra-uterina e a tradicional. As fêmeas foram distribuídas em dois tratamentos: T1 – inseminação intra-uterina com doses inseminantes contendo 0,5 bilhão de espermatozóides em um volume total de 20 ml; T2 - inseminação tradicional com doses inseminantes contendo 3,0 bilhões de espermatozóides em um volume total de 90 ml. As fêmeas receberam múltiplas inseminações. Foi possível a realização da inseminação intra-uterina em 98,1% das fêmeas. A presença de sangue, na extremidade do cateter ou espiral da pipeta de inseminação intra-uterina, foi observada em 8,4 % das fêmeas. As taxas de prenhez e de parto ajustada não diferiram entre os tratamentos. No entanto, o tamanho da leitegada foi menor (P<0,05) na IAU quando comparado à tradicional. Na inseminação artificial tradicional, em leitoas, é possível utilizar 1,5 bilhão de espermatozóides sem que as taxas de prenhez, número de embriões totais e sobrevivência embrionária sejam comprometidas. Para a inseminação intra-uterina, a utilização de 1 bilhão de espermatozóides não compromete o desempenho reprodutivo.

Aspectos clínicos e epidemiológicos da infecção genital pelo Papilomavírus humano (HPV) em adolescentes da região metropolitana de Belém

A adolescência é um período de vida da mulher marcado por transformações biológicas e psicológicas que influenciam intensamente sua saúde futura. Essas mulheres são mais suscetíveis às DST. A infecção pelo HPV é uma das DST mais frequentes, sendo importante avaliar a sua prevalência, devido sua ligação com o câncer uterino. Este estudo avaliou a prevalência da infecção genital por HPV na população adolescente do sexo feminino em Belém. Estes dados foram correlacionados com fatores sócio demográficos, comportamentais e reprodutivos. Foi realizado um estudo transversal entre agosto de 2009 e agosto de 2011, com 134 mulheres entre 13 e 19 anos que procuraram a Unidade Materno-Infantil e Adolescente de Belém para exame de rastreamento do câncer do colo do útero. As pacientes selecionadas responderam a um questionário sobre os dados sócio demográficos, comportamentais e reprodutivos. Foi realizada coleta de material cervicovaginal para citologia convencional e escovado cervical para detecção de DNA-HPV por técnica da reação em cadeia de polimerase (PCR). A associação da infecção por HPV e fatores de risco selecionados foi avaliada por meio do teste do Qui-quadrado (χ²) e/ou exato de Fisher, todos com um nível alfa de significância de 0,05. A infecção genital pelo Papillomavirus humano apresentou a prevalência de 22%, sendo o HPV 58 o mais prevalente com 31% e o HPV 11 o menos comum com 3,4%. Entre as adolescentes 51,7% apresentaram infecção por outros tipos de HPV não incluídos na vacina quadrivalente (6, 11, 16 e 18), e 76,2% apresentavam infecção por pelo menos um tipo de HPV do grupo de alto risco. Foram encontradas alterações citológicas em 6,2% dos esfregaços cervicais. Os fatores de risco associados à infecção genital por HPV encontrados neste estudo foram escolaridade superior a oito anos, coitarca com idade maior que 14 anos, uso de anticoncepcional oral por mais de um ano, uso atual de anticoncepcional oral, gravidez com 14 anos ou menos e achado citológico anormal. Este estudo evidenciou o predomínio de HPV de alto risco não imunoprevenível nas amostras cervicais, demonstrando a necessidade de novas políticas de prevenção primária e secundária, que envolvam as adolescentes através de discussão e orientação motivando-as a participar ativamente na promoção da própria saúde.

Chlamydial infection and spatial ascension of the female genital tract : a novel hybrid cellular automata and continuum mathematical model

Sexually transmitted chlamydial infection initially establishes in the endocervix in females, but if the infection ascends the genital tract, significant disease, including infertility, can result. Many of the mechanisms associated with chlamydial infection kinetics and disease ascension are unknown. We attempt to elucidate some of these processes by developing a novel mathematical model, using a cellular automata–partial differential equation model. We matched our model outputs to experimental data of chlamydial infection of the guinea-pig cervix and carried out sensitivity analyses to determine the relative influence of model parameters. We found that the rate of recruitment and action of innate immune cells to clear extracellular chlamydial particles and the rate of passive movement of chlamydial particles are the dominant factors in determining the early course of infection, magnitude of the peak chlamydial time course and the time of the peak. The rate of passive movement was found to be the most important factor in determining whether infection would ascend to the upper genital tract. This study highlights the importance of early innate immunity in the control of chlamydial infection and the significance of motility-diffusive properties and the adaptive immune response in the magnitude of infection and in its ascension.

Effects of inoculating dose on the kinetics of Chlamydia muridarum genital infection in female mice

Chlamydia trachomatis infections have been implicated in problems such as pelvic inflammatory disease and infertility in females. Although there are some studies examining the kinetics of ascending infection, there is limited information on the kinetics of pathology development and cellular infiltrate into the reproductive tissues in relation to the effects of inoculating dose, and a better understanding of these is needed. The murine model of female genital tract Chlamydia muridarum infection is frequently used as a model of human C. trachomatis reproductive tract infection. To investigate the kinetics of ascending genital infection and associated pathology development, female BALB/c mice were intravaginally infected with C. muridarum at doses ranging from 5102 to 2.6106 inclusion forming units. We found that the inoculating dose affects the course of infection and the ascension of bacteria, with the highest dose ascending rapidly to the oviducts. By comparison, the lowest dose resulted in the greatest bacterial load in the lower reproductive tract. Interestingly, we found that the dose did not significantly affect inflammatory cell infiltrate in the various regions. Overall, this data show the effects of infectious dose on the kinetics of ascending chlamydial infection and associated inflammatory infiltration in BALB/c mice.

CTA1-DD is an effective adjuvant for targeting anti-chlamydial immunity to the murine genital mucosa

Chlamydia trachomatis is a significant human pathogen with potentially severe disease sequelae in the genital tract, including infertility. A successful vaccine will need to effectively target immunity to the genital mucosa. Intranasal immunisation with cholera toxin (CT) can target immunity to the genital tract, but has the potential to cause neurological side effects. CTA1-DD is a non-toxic potent mucosal adjuvant which combines the enzymatic properties of CT, with a B cell targeting moiety. Here, we demonstrate that intranasal immunisation with CTA1-DD and chlamydial Major Outer Membrane Protein (MOMP) results in the induction of neutralising systemic and mucosal antibodies, and reduces the level of chlamydial shedding following intravaginal challenge with Chlamydia muridarum. Thus, CTA1-DD is an effective adjuvant for vaccine development against Chlamydia trachomatis, and possibly also a range of other genital pathogens.