Bacterial lipopolysaccharide induces apoptosis in the trout ovary

BACKGROUND: In mammals it is well known that infections can lead to alterations in reproductive function. As part of the innate immune response, a number of cytokines and other immune factors is produced during bacterial infection or after treatment with lipopolysaccharide (LPS) and acts on the reproductive system. In fish, LPS can also induce an innate immune response but little is known about the activation of the immune system by LPS on reproduction in fish. Therefore, we conducted studies to examine the in vivo and in vitro effects of lipopolysaccharide (LPS) on the reproductive function of sexually mature female trout. METHODS: In saline- and LPS -injected brook trout, we measured the concentration of plasma steroids as well as the in vitro steroidogenic response (testosterone and 17alpha-hydroxyprogesterone) of ovarian follicles to luteinizing hormone (LH), the ability of 17alpha,20beta-dihydroxy-4-pregnen-3-one to induce germinal vesicle breakdown (GVBD) in vitro, and that of epinephrine to stimulate follicular contraction in vitro. We also examined the direct effects of LPS in vitro on steroid production, GVBD and contraction in brook trout ovarian follicles. The incidence of apoptosis was evaluated by TUNEL analysis. Furthermore, we examined the gene expression pattern in the ovary of saline- and LPS-injected rainbow trout by microarray analysis. RESULTS: LPS treatment in vivo did not affect plasma testosterone concentration or the basal in vitro production of steroids, although a small but significant potentiation of the effects of LH on testosterone production in vitro was observed in ovarian follicles from LPS-treated fish. In addition, LPS increased the plasma concentration of cortisol. LPS treatment in vitro did not affect the basal or LH-stimulated steroid production in brook trout ovarian follicles. In addition, we did not observe any effects of LPS in vivo or in vitro on GVBD or follicular contraction. Therefore, LPS did not appear to impair ovarian steroid production, oocyte final maturation or follicular contraction under the present experimental conditions. Interestingly, LPS administration in vivo induced apoptosis in follicular cells, an observation that correlated with changes in the expression of genes involved in apoptosis, as evidenced by microarray analysis. CONCLUSION: These results indicate that female trout are particularly resistant to an acute administration of LPS in terms of ovarian hormone responsiveness. However, LPS caused a marked increase in apoptosis in follicular cells, suggesting that the trout ovary could be sensitive to the pro-apoptotic effects of LPS-induced inflammatory cytokines.

Cellular and molecular evidence for a role of tumor necrosis factor alpha in the ovulatory mechanism of trout

Background: The relevance of immune-endocrine interactions to the regulation of ovarian function in teleosts is virtually unexplored. As part of the innate immune response during infection, a number of cytokines such as tumor necrosis factor alpha (TNF alpha) and other immune factors, are produced and act on the reproductive system. However, TNF alpha is also an important physiological player in the ovulatory process in mammals. In the present study, we have examined for the first time the effects of TNF alpha in vitro in preovulatory ovarian follicles of a teleost fish, the brown trout (Salmo trutta). Methods: To determine the in vivo regulation of TNF alpha expression in the ovary, preovulatory brook trout (Salvelinus fontinalis) were injected intraperitoneally with either saline or bacterial lipopolysaccharide (LPS). In control and recombinant trout TNF alpha (rtTNF alpha)-treated brown trout granulosa cells, we examined the percentage of apoptosis by flow cytometry analysis and cell viability by propidium iodide (PI) staining. Furthermore, we determined the in vitro effects of rtTNF alpha on follicle contraction and testosterone production in preovulatory brown trout ovarian follicles. In addition, we analyzed the gene expression profiles of control and rtTNF alpha-treated ovarian tissue by microarray and real-time PCR (qPCR) analyses. Results: LPS administration in vivo causes a significant induction of the ovarian expression of TNF alpha. Treatment with rtTNF alpha induces granulosa cell apoptosis, decreases granulosa cell viability and stimulates the expression of genes known to be involved in the normal ovulatory process in trout. In addition, rtTNF alpha causes a significant increase in follicle contraction and testosterone production. Also, using a salmonid-specific microarray platform (SFA2.0 immunochip) we observed that rtTNF alpha induces the expression of genes known to be involved in inflammation, proteolysis and tissue remodeling. Furthermore, the expression of kallikrein, TOP-2, serine protease 23 and ADAM 22, genes that have been postulated to be involved in proteolytic and tissue remodeling processes during ovulation in trout, increases in follicles incubated in the presence of rtTNF alpha. Conclusions In view of these results, we propose that TNF alpha could have an important role in the biomechanics of follicle weakening, ovarian rupture and oocyte expulsion during ovulation in trout, primarily through its stimulation of follicular cell apoptosis and the expression of genes involved in follicle wall proteolysis and contraction.

Nuclear factor I-C regulates TGF-{beta}-dependent hair follicle cycling.

Skin appendages such as teeth and hair share several common signaling pathways. The nuclear factor I C (NFI-C) transcription factor has been implicated in tooth development, but a potential role in hair growth had not been assessed. In this study we found that NFI-C regulates the onset of the hair growth cycle. NFI-C(-/-) mice were delayed in the transition from the telogen to anagen phase of the hair follicle cycle after either experimental depilation or spontaneous hair loss. Lack of NFI-C resulted in delayed induction of the sonic hedgehog, Wnt5a, and Lef1 gene expression, which are key regulators of the hair follicle growth initiation. NFI-C(-/-) mice also showed elevated levels of transforming growth factor β1 (TGF-β1), an inhibitor of keratinocyte proliferation, and of the cell cycle inhibitor p21 at telogen. Reduced expression of Ki67, a marker of cell proliferation, was noted at the onset of anagen, indicating impaired activation of the hair progenitor cells. These findings implicate NFI-C in the repression of TGF-β1 signaling during telogen stage, resulting in the delay of progenitor cell proliferation and hair follicle regeneration in NFI-C-deficient mice. Taken together with prior observations, these findings also designate NFI-C as a regulator of adult progenitor cell proliferation and of postnatal tissue growth or regeneration.

Familial occurrence of an association of multiple intestinal atresia and choanal atresia: a new syndrome?

We report on two familial cases from a non-consanguineous marriage, presenting multiple intestinal and choanal atresia. Massive hydramnios and dilatation of the bowel were observed at 29 weeks of gestation during routine ultrasound scan of a healthy mother. The fetal karyotype was normal and cystic fibrosis screening was negative. Regular scans were performed throughout the pregnancy. The child was born at 34 weeks gestation. Choanal atresia was diagnosed at birth and abdominal investigations showed multiple atresia interesting both the small bowel and the colon. Further interventions were necessary because of recurrent obstructions. During the following pregnancy, a dilatation of the fetal intestinal tract was detected by ultrasonography at 27 weeks of gestation. Pregnancy was interrupted. Post-mortem examination of the fetus confirmed the stenosis of long segments of the small intestine associated with areas of colonic atresia. In both cases, histology and distribution were consistent with those reported in hereditary multiple intestinal atresia (HMIA). An association between multiple intestinal and choanal atresia has never been reported. We suggest it could correspond to a new autosomal recessive entity for which cytogenetic investigations and high-resolution array CGH revealed no visible anomalies.

Epithelium-mesenchyme interactions control the activity of peroxisome proliferator-activated receptor beta/delta during hair follicle development.

Hair follicle morphogenesis depends on a delicate balance between cell proliferation and apoptosis, which involves epithelium-mesenchyme interactions. We show that peroxisome proliferator-activated receptor beta/delta (PPARbeta/delta) and Akt1 are highly expressed in follicular keratinocytes throughout hair follicle development. Interestingly, PPARbeta/delta- and Akt1-deficient mice exhibit similar retardation of postnatal hair follicle morphogenesis, particularly at the hair peg stage, revealing a new important function for both factors in the growth of early hair follicles. We demonstrate that a time-regulated activation of the PPARbeta/delta protein in follicular keratinocytes involves the up-regulation of the cyclooxygenase 2 enzyme by a mesenchymal paracrine factor, the hepatocyte growth factor. Subsequent PPARbeta/delta-mediated temporal activation of the antiapoptotic Akt1 pathway in vivo protects keratinocytes from hair pegs against apoptosis, which is required for normal hair follicle development. Together, these results demonstrate that epithelium-mesenchyme interactions in the skin regulate the activity of PPARbeta/delta during hair follicle development via the control of ligand production and provide important new insights into the molecular biology of hair growth.

Traducción médica comentada inglés-español: Polycystic Ovary Syndrome(Joyce King)

El trabajo se divide en tres partes: una traducción del ámbito médico inglés-español; un glosario de términos extraídos del texto con su equivalente en español, y un análisis de los principales problemas de traducción detectado y que son frecuentes en los textos biomédicos, con sus respectivas soluciones.

Selection of internal control genes for real-time quantitative PCR in ovary and uterus of sows across pregnancy

Reproductive traits play a key role in pig production in order to reduce costs and increase economic returns. Among others, gene expression analyses represent a useful approach to study genetic mechanisms underlying reproductive traits in pigs. The application of reverse-transcription quantitative PCR requires the selection of appropriate reference genes, whose expression levels should not be affected by the experimental conditions, especially when comparing gene expression across different physiological stages.

Aspectos radiológicos da atresia brônquica: relato de três casos e revisão da literatura

A atresia brônquica é uma anomalia congênita rara que usualmente produz massa justa-hilar com hipertransparência distal à radiografia simples. A maioria dos pacientes é jovem e não apresenta sintomas. A tomografia computadorizada confirma o diagnóstico, permitindo o manejo conservador nos casos assintomáticos. Os autores relatam três casos de atresia brônquica. Um caso, de particular interesse, estava associado a volumosa broncocele em paciente do sexo feminino, de 45 anos de idade, que apresentou quadro sintomático de infecção respiratória. O diagnóstico foi estabelecido por meio dos achados da radiologia convencional, da tomografia computadorizada e da comparação com radiografias convencionais e tomografia linear prévios, que já demonstravam alterações. Realizou-se uma revisão sobre os achados clínicos e radiológicos dessa malformação.

Atresia do trato gastrintestinal: avaliação por métodos de imagem

Um amplo espectro de anomalias congênitas pode afetar qualquer nível do trato gastrintestinal, do esôfago ao ânus. A atresia é uma importante causa de obstrução gastrintestinal, com alta taxa de morbidade em recém-natos. Há diversos mecanismos patológicos possíveis para explicar esta malformação e duas explicações clássicas de sua gênese são um defeito de recanalização do tubo intestinal ou uma interrupção no suprimento sanguíneo durante a vida intra-uterina. Os autores fazem uma revisão da literatura com ensaio iconográfico dos achados de imagem em crianças com atresia do trato gastrintestinal.

Unilateral pulmonary veins atresia: evaluation by computed tomography

Unilateral pulmonary vein atresia is a rare congenital condition. In addition to cardiac malformations or pulmonary hypertension, patients may present with recurrent pulmonary infections or hemoptysis in childhood or adolescence. The authors report a case where the typical findings of such condition were observed at computed tomography in an adult patient.

Imaging findings of bronchial atresia in fetuses, neonates and infants.

Congenital lung malformations are increasingly detected before birth. However, bronchial atresia is rarely identified in utero and not always recognized in neonates. There are two types of atresia: 1) proximal, located at the level of the mainstem or the proximal lobar bronchi, which is extremely rare and usually lethal during pregnancy, causing a tremendous volume increase of the distal involved lung with secondary hypoplasia of the normal lung, and 2) peripheral, located at the segmental/subsegmental bronchial level, which may present as an isolated lesion or as part of a complex congenital malformation. Prenatal findings are mostly nonspecific. Postnatal exams show overinflated lung areas and focal bronchial dilations. The typical fluid-filled bronchoceles are not always observed in neonates but develop progressively in the first months of life. This pictorial essay describes the spectrum of imaging findings of bronchial atresia in fetuses, neonates and infants.

Tratamento cirúrgico da atresia de vias biliares: a experiência do hospital municipal jesus, 1997-2000

OBJETIVO: Analisar a indicação, resultados e limitações da portoenterostomia para o tratamento da atresia de vias biliares em hospital infantil terciário no Rio de Janeiro-RJ/Brasil. MÉTODO: Foram estudados prospectivamente oito pacientes submetidos à portoenterostomia do tipo Kasai 1, num período de três anos (1997-2000). Todos foram submetidos à antibioticoterapia profilática, uso de colerético, reavaliação médica mensal nos primeiros seis meses de seguimento e reavaliação laboratorial, ultra-sonográfica e através de endoscopia digestiva alta em prazos programados. RESULTADOS: Em cinco pacientes foi possível obter drenagem biliar estável e satisfatória por > 6 meses pós-operatórios. Quatro faleceram no período de observação (um por insuficiência hepática, um por colangite, dois por hemorragias digestivas, inclusive os três pacientes em que não foi obtida drenagem biliar satisfatória). Quatro persistem em bom estado geral, anictéricos, um deles com fibrose hepática residual grave e provável indicação futura de transplante hepático. Detectamos problemas graves com relação ao encaminhamento tardio de doentes para tratamento e para disponibilizar transplante hepático quando necessário. CONCLUSÕES: Os resultados da portoenterostomia são compensadores quando é possível obter bom débito biliar no pós-operatório. Em pacientes em que não é obtida boa drenagem biliar o pós-operatório é tormentoso e o óbito é esperado até o segundo ano de vida sem o uso de transplante hepático. É necessário disponibilizar transplante hepático pediátrico em nosso meio e conscientizar a comunidade médica para o encaminhamento precoce de bebês ictéricos além dos primeiros 15 dias de vida para investigação e tratamento em unidades especializadas.

Polycystic ovary syndrome: implications of metabolic dysfunction

OBJECTIVE: To determine the prevalence of metabolic syndrome (MS) and its clinical interrelations in polycystic ovary syndrome (PCOS).METHODS: This was a cross-sectional, prospective study with 100 patients with diagnosed PCOS based on the consensus of Rotterdam (2003). We investigated the interrelationships of MS, with intrinsic PCOS data. Dermatological profile was analyzed, in addition to acanthosis nigricans (AN) in the presence of hirsutism and acne. The use of HOMA-IR (homeostatic model assessment of insulin resistance) aimed at the correlation with MS in order to establish the metabolic dysfunction with the state of insulin resistance.RESULTS: The mean and standard deviations corresponding figures for age, body mass index and waist circumference were, respectively, 25.72 (± 4.87), 30.63 (± 9.31) and 92.09 (± 18.73). The prevalence of MS was 36% and significantly correlated with BMI, AN, and in 51% of patients the state of insulin resistance (HOMA-IR). Regarding skin profile, only AN significant correlation with MS.CONCLUSION: We propose the routine inspection of metabolic components related to severe PCOS. These parameters configure the cardiovascular risk and such conduct is of undoubted importance to public health.