Plasma concentrations and placental immunostaining of interleukin-10 and tumornecrosis factor-α as predictors of alterations in the embryo-fetal organism and the placental development of diabetic rats

Interleukin-10 (IL-10) appears to be the key cytokine for the maintenance of pregnancy and inhibits the secretion of inflammatory cytokines such as tumor necrosis factor-α (TNF-α). However, there are no studies evaluating the profile of these cytokines in diabetic rat models. Thus, our aim was to analyze IL-10 and TNF-α immunostaining in placental tissue and their respective concentrations in maternal plasma during pregnancy in diabetic rats in order to determine whether these cytokines can be used as predictors of alterations in the embryo-fetal organism and in placental development. These parameters were evaluated in non-diabetic (control; N = 15) and Wistar rats with streptozotocin (STZ)-induced diabetes (N = 15). At term, the dams (100 days of life) were killed under anesthesia and plasma and placental samples were collected for IL-10 and TNF-α determinations by ELISA and immunohistochemistry, respectively. The reproductive performance was analyzed. Plasma IL-10 concentrations were reduced in STZ rats compared to controls (7.6 ± 4.5 vs 20.9 ± 8.1 pg/mL). The placental scores of immunostaining intensity did not differ between groups (P > 0.05). Prevalence analysis showed that the IL-10 expression followed TNF-α expression, showing a balance between them. STZ rats also presented impaired reproductive performance and reduced plasma IL-10 levels related to damage during early embryonic development. However, the increased placental IL-10 as a compensatory mechanism for the deficit of maternal regulation permitted embryo development. Therefore, the data suggest that IL-10 can be used as a predictor of changes in the embryo-fetal organism and in placental development in pregnant diabetic rats.

Triorganotin as a compound with potential reproductive toxicity in mammals

Organotin compounds are typical environmental contaminants and suspected endocrine-disrupting substances, which cause irreversible sexual abnormality in female mollusks, called "imposex". However, little is known about the capability of triorganotin compounds, such as tributyltin and triphenyltin, to cause disorders in the sexual development and reproductive functions of mammals, including humans and rodents. Moreover, these compounds can act as potential competitive inhibitors of aromatase enzyme and other steroidogenic enzymes, affecting the reproductive capacity of male and female mammals. In this review, we discuss the cellular, biochemical, and molecular mechanisms by which triorganotin compounds induce adverse effects in the mammalian reproductive function.

Family rivalry in the testis: Retinoblastoma protein and E2F transcription factors in testicular development and adult spermatogenesis

Disorders of male reproductive health are becoming increasingly prevalent globally. These defects, ranging from decreasing sperm counts to an increasing rate of infertility and testicular cancer, have a common origin in the early phases of testicular development, but the exact mechanisms that cause them remain unknown. Testicular development and adult spermatogenesis are complex processes in which different cell types undergo mitosis, meiosis, differentiation and apoptosis. The retinoblastoma protein family and its associated E2F transcription factors are key regulators of these cellular events. In the present study, the functions of these factors in postnatal testicular development and adult spermatogenesis were explored using different animal models. In addition, a new application of flow cytometry to study testicular cell dynamics was developed. An ablation of retinoblastoma protein in mouse Sertoli cells resulted in their cell cycle re-entry in adult testes, dedifferentiation and a severe spermatogenic defect. We showed that deregulated E2F3 contributed to these changes. Our results indicated that the E2F1 transcription factor is critical for the control of apoptosis in the developing postnatal testis. In the adult testis, E2F1 controls the maintenance of the spermatogonial stem cell pool, in addition to inhibiting apoptosis of spermatocytes. In summary, this study elucidated the complex interdependencies of the RB and E2F transcription factor families in the control of postnatal testicular development and adult spermatogenesis. Furthermore, this study provided a new methodology for the analysis of testicular cells.

RR soybean seed quality after application of glyphosate in different stages of crop development

In consequence of several studies and speculations concerning the issue of RR transgenic soybean after the application of glyphosate, additional scientific investigations became necessary to clarify the actual viability of the product use when applied in different developmental stages of the soybean crop. Therefore, this study was aimed to evaluate the physiological quality as well as seed health quality of RR soybean subjected to application of the herbicide glyphosate in different phonological stages of the transgenic soybean, cultivar CD 219RR. For this, an experiment with a complete block experimental design with treatments randomly distributed within the block, with four replications, was carried out. The assessed treatments were foliar sprayings of glyphosate in three increasing dosages [0 (control); 1,440 g ha-1; and 2,880 g ha-1] of acid equivalent, applied in two crop developmental stages: vegetative (V6) and reproductive (R2). The variables assessed were: germination; first count of germination; fresh and dry mass of seedlings, lengths of seedling and root; vigor and viability by the tetrazolium test; and seed health quality. Glyphosate application may adversely affect physiological quality of RR soybean seeds, when applied in dosages varying from 1,440 to 2,880 g acid equivalent per hectare at the stages V6 and R2.

Actinobacillus pleuropneumoniae Possesses an Antiviral Activity against Porcine Reproductive and Respiratory Syndrome Virus

Pigs are often colonized by more than one bacterial and/or viral species during respiratory tract infections. This phenomenon is known as the porcine respiratory disease complex (PRDC). Actinobacillus pleuropneumoniae (App) and porcine reproductive and respiratory syndrome virus (PRRSV) are pathogens that are frequently involved in PRDC. The main objective of this project was to study the in vitro interactions between these two pathogens and the host cells in the context of mixed infections. To fulfill this objective, PRRSV permissive cell lines such as MARC-145, SJPL, and porcine alveolar macrophages (PAM) were used. A pre-infection with PRRSV was performed at 0.5 multiplicity of infection (MOI) followed by an infection with App at 10 MOI. Bacterial adherence and cell death were compared. Results showed that PRRSV preinfection did not affect bacterial adherence to the cells. PRRSV and App co-infection produced an additive cytotoxicity effect. Interestingly, a pre-infection of SJPL and PAM cells with App blocked completely PRRSV infection. Incubation of SJPL and PAM cells with an App cell-free culture supernatant is also sufficient to significantly block PRRSV infection. This antiviral activity is not due to LPS but rather by small molecular weight, heat-resistant App metabolites (,1 kDa). The antiviral activity was also observed in SJPL cells infected with swine influenza virus but to a much lower extent compared to PRRSV. More importantly, the PRRSV antiviral activity of App was also seen with PAM, the cells targeted by the virus in vivo during infection in pigs. The antiviral activity might be due, at least in part, to the production of interferon c. The use of in vitro experimental models to study viral and bacterial co-infections will lead to a better understanding of the interactions between pathogens and their host cells, and could allow the development of novel prophylactic and therapeutic tools.

Identification of a new cell line permissive to porcine reproductive and respiratory syndrome virus infection and replication which is phenotypically distinct from MARC-145 cell line

Background Airborne transmitted pathogens, such as porcine reproductive and respiratory syndrome virus (PRRSV), need to interact with host cells of the respiratory tract in order to be able to enter and disseminate in the host organism. Pulmonary alveolar macrophages (PAM) and MA104 derived monkey kidney MARC-145 cells are known to be permissive to PRRSV infection and replication and are the most studied cells in the literature. More recently, new cell lines developed to study PRRSV have been genetically modified to make them permissive to the virus. The SJPL cell line origin was initially reported to be epithelial cells of the respiratory tract of swine. Thus, the goal of this study was to determine if SJPL cells could support PRRSV infection and replication in vitro. Results The SJPL cell growth was significantly slower than MARC-145 cell growth. The SJPL cells were found to express the CD151 protein but not the CD163 and neither the sialoadhesin PRRSV receptors. During the course of the present study, the SJPL cells have been reported to be of monkey origin. Nevertheless, SJPL cells were found to be permissive to PRRSV infection and replication even if the development of the cytopathic effect was delayed compared to PRRSV-infected MARC-145 cells. Following PRRSV replication, the amount of infectious viral particles produced in SJPL and MARC-145 infected cells was similar. The SJPL cells allowed the replication of several PRRSV North American strains and were almost efficient as MARC-145 cells for virus isolation. Interestingly, PRRSV is 8 to 16 times more sensitive to IFNα antiviral effect in SJPL cell in comparison to that in MARC-145 cells. PRRSV induced an increase in IFNβ mRNA and no up regulation of IFNα mRNA in both infected cell types. In addition, PRRSV induced an up regulation of IFNγ and TNF-α mRNAs only in infected MARC-145 cells. Conclusions In conclusion, the SJPL cells are permissive to PRRSV. In addition, they are phenotypically different from MARC-145 cells and are an additional tool that could be used to study PRRSV pathogenesis mechanisms in vitro.

Development and use of a multiplex real-time quantitative polymerase chain reaction assay for detection and differentiation of Porcine circovirus-2 genotypes 2a and 2b in an epidemiological survey.

By the end of 2004, the Canadian swine population had experienced a severe 2 increase in the incidence of Porcine circovirus-associated disease (PCVAD), a problem that was 3 associated with the emergence of a new Porcine circovirus-2 genotype (PCV-2b), previously 4 unrecovered in North America. Thus it became important to develop a diagnostic tool that could 5 differentiate between the old and new circulating genotypes (PCV-2a and -2b, respectively). 6 Consequently, a multiplex real-time quantitative polymerase chain reaction (mrtqPCR) assay that 7 could sensitively and specifically identify and differentiate PCV-2 genotypes was developed. A 8 retrospective epidemiological survey that used the mrtqPCR assay was performed to determine if 9 cofactors could affect the risk of PCVAD. From 121 PCV-2–positive cases gathered for this 10 study, 4.13%, 92.56% and 3.31% were positive for PCV-2a, PCV-2b, and both genotypes, 11 respectively. In a data analysis using univariate logistic regressions, PCVAD compatible 12 (PCVAD/c) score was significantly associated with the presence of Porcine reproductive and 13 respiratory syndrome virus (PRRSV), PRRSV viral load, PCV-2 viral load, and PCV-2 14 immunohistochemistry (IHC) results. Polytomous logistic regression analysis revealed that 15 PCVAD/c score was affected by PCV-2 viral load (P = 0.0161) and IHC (P = 0.0128), but not by 16 the PRRSV variables (P > 0.9); suggesting that mrtqPCR in tissue is a reliable alternative to IHC. 17 Logistic regression analyses revealed that PCV-2 increased the odds ratio of isolating 2 major 18 swine pathogens of the respiratory tract, Actinobacillus pleuropneumoniae and Streptococcus 19 suis serotypes 1/2, 1, 2, 3, 4, and 7, which are serotypes commonly associated with clinical 20 diseases.

Actinobacillus pleuropneumoniae induces SJPL cell cycle arrest in G2/M-phase and inhibits porcine reproductive and respiratory syndrome virus replication

Background: Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important pathogens in the swine industry and causes important economic losses. No effective antiviral drugs against it are commercially available. We recently reported that the culture supernatant of Actinobacillus pleuropneumoniae, the porcine pleuropneumonia causative agent, has an antiviral activity in vitro against PRRSV in SJPL cells. Objectives of this study were (i) to identify the mechanism behind the antiviral activity displayed by A. pleuropneumoniae and (ii) to characterize the active molecules present in the bacterial culture supernatant. Methods: Antibody microarray analysis was used in order to point out cellular pathways modulated by the A. pleuropneumoniae supernatant. Subsequent, flow cytometry analysis and cell cycle inhibitors were used to confirm antibody microarray data and to link them to the antiviral activity of the A. pleuropneumoniae supernatant. Finally, A. pleuropneumoniae supernatant characterization was partially achieved using mass spectrometry. Results: Using antibody microarray, we observed modulations in G2/M-phase cell cycle regulation pathway when SJPL cells were treated with A. pleuropneumoniae culture supernatant. These modulations were confirmed by a cell cycle arrest at the G2/M-phase when cells were treated with the A. pleuropneumoniae culture supernatant. Furthermore, two G2/M-phase cell cycle inhibitors demonstrated the ability to inhibit PRRSV infection, indicating a potential key role for PRRSV infection. Finally, mass spectrometry lead to identify two molecules (m/z 515.2 and m/z 663.6) present only in the culture supernatant. Conclusions: We demonstrated for the first time that A. pleuropneumoniae is able to disrupt SJPL cell cycle resulting in inhibitory activity against PRRSV. Furthermore, two putative molecules were identified from the culture supernatant. This study highlighted the cell cycle importance for PRRSV and will allow the development of new prophylactic or therapeutic approaches against PRRSV.

Studies on the reproductive physiology of the green tiger prawn Penaeus (penaeus) semisulcatus de Haan

A study of the reproductive physiology of P. (P). semisulcatus wasundertaken as this information is an essential prerequisite for brood stock development for hatchery operations, and the results are embodied in this thesis. The thesis is presented in seven chapters. The protein, lipid and carbohydrate contents of ovary were estimated for the different vitellogenic phases. The protein, lipid, carbohydrate and cholesterol contents were estimated in testes and spermatophore.The thesis present the results of induced maturation experiments by eyestalk ablation, CNS extract injection and UV rays application on immature female prawns.

Studies on Reproductive Biology euui Mass Larval Rearing of Selected Species of Macrobrach

In India scientific commercial farming of Macrobrachium is yet to get a start, the major constraint being the lack of seed. Seed collection from the natural sources alone may not be sufficient, and in such cases seed production in hatcheries is inevitable. This necessitates knowledge of the reproductive biology of the species technology development for brood stock and larval rearing, and the optimum conditions required for larval development to undertake large scale seed production.The present study was taken up with the major objectives of developing a proper, simple and feasible technology for the larval rearing of Macrobrachium, with special emphasis on. mass larval rearing suitable for commercial level operations.

Studies on the reproductive endocrinology of the penaeid prawn, Penaeus indicus H. Milne Edward

The present investigation has been carried out to understand the process and events leading to maturation of the ovary and testis in the Indian white prawn Penaeus indicus. The study includes the classification of the ovarian maturity stages based on its colour, gonadosomatic index, oocyte diameter and morphological changes in the oocyte. Further the process of oogenesis has been investigated using light and electron microscopic techniques. A histochemical study of the ovary has also been carried out to determine the sequence in which yolk substances are synthesized or sequestered in the oocytes and also to elicit the nature of the penaeid yolk material. The process of spermatogenesis and the development of the spermatophore has been studied in detail using light and electron microscopic methods. In addition a brief histochemical study on the testis was also made to understand the nature of the organic reserves in the sperm cells.

Studies on Selected Myctophid Fishes in Arabian Sea with respect to their Status in Deep Sea Trawl Bycatch, Length-Weight Relationship, Reproductive Biology and Population Dynamics

Available information on abundance of myctophids and their utilisation indicate that there is excellent scope for development of myctophid fisheries in Indian Ocean. Most of the conventional fish stocks have reached a state of full exploitation or over-exploitation. Hence there is need to locate new and conventional fishery resources in order to fill in the supply-demand gap, in the face of increasing demand for fish. Information on length-weight relationship, age and growth, spawning season, fecundity and age at maturity and information on bycatch discards are required for sustainable utilization of myctophid resource in the Indian Ocean

Teleconsultation in sexual and reproductive health for young adults through mobile devices: Experience in Colombia

Background: Sexual risk behaviors associated with poor information on sexuality have contributed to major public health problems in the area of sexual and reproductive health in teenagers and young adults in Colombia. Objective: To measure the perception of changes in sexual and reproductive risk behavior after the use of a teleconsultation service via mobile devices in a sample of young adults. Methods: A before and after observational study was designed, where a mobile application to inquire about sexual and reproductive health was developed. The perception of changes in sexual and reproductive health risk behaviors in a sample of young adults after the use of the application was measured using the validated survey “Family Health International (FHI) – Behavioral Surveillance Survey (BSS) – Survey for Adults between 15 to 40 Years”. Non-probabilistic convenience recruitment was undertaken through the study´s web page. Participants answered the survey online before and after the use of the mobile application for a six month period (intervention). For the inferential analysis, data was divided into three groups (dichotomous data, discrete quantitative data, and ordinal data), to compare the results of the questions between the first and the second survey. For all tests, a confidence interval of 95% was established. For dichotomous data, the Chi-squared test was used. For quantitative data, we used the Student’s t-test, and for ordinal data, the Mann-Whitney-Wilcoxon test. Results: A total of 257 subjects were registered in the study and met the selection criteria. The pre-intervention survey was answered by 232 subjects, and 127 completely answered the post-intervention survey, of which 54.3% did not use the application, leaving an effective population of 58 subjects for analysis. 53% (n=31) were female, and 47% (n=27) were male. The mean age was 21 years, ranging between 18 and 40 years. The differences between the answers on the first and the second survey were not statistically significant. The main risk behaviors identified in the population were homosexual relations, non-use of condoms, sexual relations with non-regular and commercial partners, the use of psychoactive substances, and ignorance about the symptoms of sexually transmitted diseases and HIV transmission. Conclusions: Although there were no differences between the pre- and post-intervention results, the study revealed different risk behaviors among the participating subjects. These findings highlight the importance of promoting educational strategies on this matter and the importance of providing patients with easily accessible tools with reliable health information.

Equine conceptus development : a mini review

Many aspects of early embryonic development in the horse are unusual or unique; this is of scientific interest and, in some cases, considerable practical significance. During early development the number of different cell types increases rapidly and the organization of these increasingly differentiated cells becomes increasingly intricate as a result of various inter-related processes that occur step-wise or simultaneously in different parts of the conceptus (i.e., the embryo proper and its associated membranes and fluid). Equine conceptus development is of practical interest for many reasons. Most significantly, following a high rate of successful fertilization (71-96%) (Ball, 1988), as many as 30-40% of developing embryos fail to survive beyond the first two weeks of gestation (Ball, 1988), the time at which gastrulation begins. Indeed, despite considerable progress in the development of treatments for common causes of sub-fertility and of assisted reproductive techniques to enhance reproductive efficiency, the need to monitor and rebreed mares that lose a pregnancy or the failure to produce a foal, remain sources of considerable economic loss to the equine breeding industry. Of course, the potential causes of early embryonic death are numerous and varied (e.g. persistent mating induced endometritis, endometrial gland insufficiency, cervical incompetence, corpus luteum (CL) failure, chromosomal, genetic and other unknown factors (LeBlanc, 2004). However, the problem is especially acute in aged mares with a history of poor fertility in which the incidence of embryonic loss between days 2 and 14 after ovulation has been reported to reach 62-73%, and in which embryonic death is due primarily to embryonic defects rather than to uterine pathology (Ball et al., 1989; Carnevale & Ginther, 1995; Ball, 2000).

Intergenerational effects of birth weight on glucose tolerance and reproductive performance

Women who were themselves small-for-gestational age (SGA) are at a greater risk of adulthood diseases such as non-insulin-dependent diabetes mellitus (NIDDM), and twice at risk of having an SGA baby themselves. The aim of this study was to examine the intergenerational pig. Low (L) and normal (N) birth weight female piglets were followed throughout their first pregnancy (generation 1 (0)). After they had given birth, the growth and development of the lightest (I) and heaviest (n) female piglet from each litter were monitored until approximately 5 months of age (generation 2 (G2)). A glucose tolerance test (GTT) was conducted on G1 pig at similar to 6 months of age and again during late pregnancy; a GTT was also conducted on G2 pigs at similar to 4 months of age. G1 L offspring exhibited impaired glucose metabolism in later life compared to their G1 N sibling but in the next generation a similar scenario was only observed between I and n offspring born to G1 L mothers. Despite G1 L mothers showing greater glucose intolerance in late pregnancy and a decreased litter size, average piglet birth weight was reduced and there was also a large variation in litter weight; this suggests that they were, to some extent, prioritising their nutrient intake towards themselves rather than promoting their reproductive performance. There were numerous relationships between body shape at birth and glucose curve characteristics in later life, which can, to some extent, be used to predict neonatal outcome. In conclusion, intergenerational effects are partly seen in the pig. It is likely that some of the intergenerational influences may be masked due to the pig being a litter-bearing species.