A study protocol of vaginal laser therapy in gynecological cancer survivors.

Objectives: Sexual dysfunction and radiation cystitis are common adverse events following radiotherapy for gynecological cancer (GC). This study aims to assess the efficacy of intravaginal CO2 laser on GC survivors with dyspareunia following pelvic radiation and/or brachytherapy.Methods: This is the study protocol of a randomized double-blind placebo-controlled trial. All participants will receive five therapies (active or placebo) at monthly intervals. Outcomes will include a 10-cm visual analog scale measuring dyspareunia, vaginal dryness, and other symptom intensity, 3-day voiding diary, Day-to Day Impact of Vaginal Aging questionnaire, Female Sexual Function Index, European Organization for Research and Treatment of Cancer Quality of Life questionnaire cervical cancer module, Urogenital Distress Inventory short form, King's Health Questionnaire, International Consultation on Incontinence Questionnaire short form/female lower urinary tract symptoms, patient perception of improvement, sexual satisfaction of male partners, vaginal maturation value, and Vaginal Health Index. Differences between groups will be assessed at baseline and 1, 3, 6, 9, and 12 months following the five laser therapies.Results: As this is a study protocol, the study is ongoing with an expected end of recruitment and analysis date of 2021.Conclusion: Pelvic radiotherapy for GC increases the 5-year survival rate but with a negative impact on women's quality of life due to sexual dysfunction and radiation cystitis onset. With this study, CO2 laser therapy will be evaluated for the first time in GC survivors treated with radiotherapy. ClinicalTrials.gov registration number: NCT03714581.

Regularized Joint Estimation of Related Vector Autoregressive Models.

In a number of applications, one has access to high-dimensional time series data on several related subjects. A motivating application area comes from the neuroimaging field, such as brain fMRI time series data, obtained from various groups of subjects (cases/controls) with a specific neurological disorder. The problem of regularized joint estimation of multiple related Vector Autoregressive (VAR) models is discussed, leveraging a group lasso penalty in addition to a regular lasso one, so as to increase statistical efficiency of the estimates by borrowing strength across the models. A modeling framework is developed that it allows for both group-level and subject-specific effects for related subjects, using a group lasso penalty to estimate the former. An estimation procedure is introduced, whose performance is illustrated on synthetic data and compared to other state-of-the-art methods. Moreover, the proposed approach is employed for the analysis of resting state fMRI data. In particular, a group-level descriptive analysis is conducted for brain inter-regional temporal effects of Attention Deficit Hyperactive Disorder (ADHD) patients as opposed to controls, with the data available from the ADHD-200 Global Competition repository.

Cytokine activation during embryonic development and in hen ovary and vagina during reproductive age and Salmonella infection.

Salmonellosis is one of the most important zoonotic diseases and is usually associated with consumption of Salmonella Enteritidis (SE) contaminated poultry meat or eggs. Contamination with SE is usually the result of infection of the digestive tract, or reproductive organs, especially the ovary and vagina. Thus, knowledge of endogenous innate immune mechanisms operating in the ovary and vagina of hen is an emerging aspect of reproductive physiology. Cytokines are key factors for triggering the immune response and inflammation in chicken to Salmonella infection. The aim of this study was to investigate the expression profile of 11 proinflammatory cytokines in the chicken embryos during embryonic development, as well as in the hen ovary and vagina in vivo, to investigate whether sexual maturation affects their ovarian and vaginal mRNA abundance and to determine whether cytokine expression was constitutive or induced in the ovary and vagina as a response to SE infection. RT-PCR analysis revealed that several cytokines were expressed in the chicken embryos, and in the ovary and vagina of healthy birds. Expression of various cytokines during sexual maturation appeared to be developmentally regulated. In addition, a significant up-regulation of several cytokines in the ovary and vagina of sexually mature SE infected birds compared to healthy birds of the same age was observed. These results suggest a cytokine-mediated immune response mechanism against Salmonella infection in the hen reproductive organs.

Transcriptional changes of cytokines in rooster testis and epididymis during sexual maturation stages and Salmonella infection.

Infection of rooster testis and epididymis by pathogens can lead to impaired fertility, resulting in economic losses in the poultry industry. Antimicrobial protection of rooster reproductive organs is, therefore, an important aspect of reproductive physiology. Salmonellosis is one of the most important zoonotic diseases, caused by Salmonella bacteria including Salmonella Enteritidis (SE) and is usually the result of infection of the reproductive organs. Thus, knowledge of the endogenous innate immune mechanisms of the rooster testis and epididymis is an emerging aspect of reproductive physiology. Cytokines are key factors for stimulating the immune response and inflammation in chickens to Salmonella infection. In the present study the expression profile of 11 pro-inflammatory cytokine genes in the rooster testis and epididymis in vivo and transcriptional changes in these organs during sexual maturation and SE infection were investigated. Gene expression analysis data revealed that in both testis and epididymis nine cytokines namely the IL-1ß, IL-6, IL-8, IL-10, IL-12, IL-15, IL-16, IL-17 and IL-18 genes were expressed, while no mRNA transcripts were detected in both organs for IL-2 and IL-4. Furthermore, the expression of various cytokine genes during sexual maturation appeared to be developmentally regulated, while SE infection resulted in a significant up-regulation of IL-1ß, -6, -12 and -18 genes in the testis and an increase in the mRNA relative abundance of IL-1ß, -6, -12, -16 and -18 in the epididymis of SE-infected sexually mature 28-week-old roosters. These results suggest a cytokine-mediated immune response mechanism against Salmonella infection in the rooster reproductive tract.

Effects of maternal undernutrition during late gestation and/or lactation on colostrum synthesis and immunological parameters in the offspring.

The emerging immune system is vulnerable to insult not only during fetal life, but also through colostrum transfer of maternal factors with immunomodulatory functions. The aim of the present study was to examine the effects of maternal undernutrition during late gestation and/or lactation on colostrum and milk synthesis, as well as on immunological parameters in offspring. Pregnant ewes were fed to 100% of nutrient requirements throughout pregnancy and lactation (Control) or to 50% during lactation (R1) or during the last 20 days of pregnancy and lactation (R2). Colostrum samples were collected 3 and 18h after parturition and thymus glands were obtained from 5-month-old offspring. Lamb birthweight did not differ between groups, whereas growth rate was significantly lower in males in the R1 group and in females in both undernourished groups. There was a significant reduction in lactose percentage in the 18-h colostrum of the R2 group. The IgG concentration, as a percentage of protein, was significantly increased in 3-h colostrum samples of the R2 group. Quantitative polymerase chain reaction analysis revealed a significant increase in the expression of Toll-like receptor (TLR) 2, TLR4 and TLR9 in the thymus gland of female lambs in both undernourished groups. In conclusion, early life nutritional imbalances may impact on immune system function in later life due to programming effects.

A multi-farm assessment of Greek black pig genetic diversity using microsatellite molecular markers.

Local breeds are important for the maintenance of genetic diversity and future food security. Nowadays, the worldwide distribution of pigs is dominated by a few breeds, tending towards a severe loss of pig biodiversity. Thus, it is critical to maintain distinct populations of pig breeds. The Greek black pig, a breed raised locally and known for the high quality of its meat for cured products, is the only traditional indigenous pig breed reared in Greece. We investigated the genetic diversity, based on microsatellite analysis, of the Greek black pig and evaluated its genetic uniqueness. One hundred and three pigs from 12 Greek farms were analyzed using 11 microsatellites. The total number of alleles amounted to 135, with a mean number of alleles per locus of 12.27, ranging between 10 and 16 alleles. The observed heterozygosity ranged from 0.363 to 0.825 per locus. The expected heterozygosity ranged from 0.471 to 0.707. The inbreeding coefficient ranged from -0.329 to 0.229. We conclude that the Greek black pig, despite its low population size, has a high degree of genetic variability, which will be useful for breeding programs aimed at maintaining long-term survival of this ancient breed.

Effects of sexual maturation and Salmonella infection on the expression of avian ß-defensin genes in the chicken testis.

Rooster infertility is a major concern in the poultry industry and protection of the male reproductive organs from pathogens is an essential aspect of reproductive physiology. During the last years, research on antimicrobial protection has elucidated the critical role of the antimicrobial peptides avian ß-defensins (AvBDs) in the innate immunity in chickens. AvBDs have been reported to be expressed in the hen reproductive organs, providing protection against microbial pathogens including Salmonella Enteritidis (SE). However, mechanisms of antimicrobial protection of rooster reproductive organs and especially the testis, mediated by AvBDs are poorly understood. The aim of this study was to investigate the expression of the complete family of the 14 AvBD genes, in the rooster testis in vivo, to determine whether sexual maturation affects their testicular mRNA abundance and to investigate whether SE infection alters their expression. Expression analysis revealed that 9 members of the AvBD family, namely AvBD1, 2, 4, 5, 6, 9, 10, 12 and 14 were expressed in the testis. Quantitative real-time PCR analysis revealed that the mRNA abundance of three AvBDs was up regulated and of three AvBDs was down regulated with respect to sexual maturation. In addition, SE infection resulted in a significant induction of AvBD4, 10, 12 and 14 in the testis of sexually mature roosters. These findings provide strong evidence to suggest that an AvBD-mediated immune response mechanism exists in the rooster testis providing protection against bacterial pathogens including Salmonella species.

Association of BF gene polymorphism with litter size in a commercial pig cross population.

In the present study, the single nucleotide polymorphism of CFB (Complement Factor B), BF gene, was analyzed by PCR-RFLP in a commercial pig population in Greece. BF gene is coding for properdin, a protein that plays an integral role in the uterine epithelium growth and is found in a QTL region with many other genes associated with specific reproductive traits. BF gene is considered to affect various reproductive traits in pigs and the present study associated the BF gene genotypes with litter size of the sows (TNB, total number born and NBA number born alive piglets/birth). Sows with AB genotype gave statistically significant lower values for TNB and NBA piglets/birth (p<0.05) than the BB genotype. Our results support the concept that BF belongs to the group of genes that control the litter size of the sows and thus BF gene could be used for Marker-assisted selection programmes for the genetic improvement of reproductive characteristics in livestock.

Expression of avian ß-defensins and Toll-like receptor genes in the rooster epididymis during growth and Salmonella infection.

The epididymis is an organ involved in the maturation, transport, and storage of sperm prior to ejaculation. As epididymis is exposed to a constant risk of inflammatory conditions that may lead to transient or permanent sterility, protection of this organ from pathogens is an essential aspect of reproductive physiology. The families of antimicrobial peptides ß-defensins and the pattern-recognition receptors Toll-like (TLR) mediate innate immunity in various vertebrates including avian species. As rooster infertility is a major concern in the poultry industry, the objectives of this study were to determine the expression profile of the entire family of the avian ß-defensins (AvBD) and TLR genes in the rooster epididymis, to investigate whether sexual maturation affects their epididymidal mRNA abundance and to determine the changes in their expression levels in response to Salmonella enteritidis (SE) infection in the epididymis of sexually mature roosters. RNA was extracted from the epididymis of healthy pubertal, sexually mature and aged birds, and from sexually mature SE infected birds. RT-PCR analysis revealed that 10 members of the AvBD and nine members of the TLR gene families were expressed in the epididymis. Quantitative real-time PCR analysis revealed that the epididymidal mRNA abundance of certain AvBD and TLR genes was developmentally regulated with respect to sexual maturation. SE infection resulted in a significant induction of AvBD 1, 9, 10, 12 and 14, as well as TLR 1-2, 2-1, 2-2, 4, 5 and 7 genes, in the epididymis of sexually mature roosters, compared to healthy birds of the same age. These findings provide strong evidence to suggest that the rooster epididymis is capable of initiating an inflammatory response to Salmonella, through activation of certain members of the AvBD and TLR gene families.

Transcriptional profiling of antimicrobial peptides avian ß-defensins in the chicken ovary during sexual maturation and in response to Salmonella enteritidis infection.

Avian ß-defensins (AvßDs) are antimicrobial peptides that play significant roles in the innate immune system in chickens. The aim of this study was to identify the types of AvßDs expressed in the chicken ovary, to investigate the effects of sexual maturation in the ovarian mRNA abundance and to determine the changes in their expression levels as a result to Salmonella enteritidis (SE) infection. RNA was extracted from the ovary of healthy prepubertal, sexually mature and aged birds, as well as from sexually mature and aged SE infected birds. Real-time PCR analysis revealed that 11 AvßDs genes were expressed in the chicken ovary. A significant up regulation of AvßD1, 3, 4, 5, 7 and 11 was observed in the ovary of sexually mature and aged birds. Furthermore, a significant up-regulation of AvßD4, 5, 7, 11 and 12 was observed in the ovary of SE infected sexually mature birds. These results suggest that the mRNA expression of at least six AvßDs increase with age in the ovary of laying hens, and that at least five AvßDs show an induction in their expression in response to SE infection, indicating an AvßD-mediated immune response mechanism in the chicken ovary.

Changes in the expression of Toll-like receptors in the chicken testis during sexual maturation and Salmonella infection.

Rooster infertility is a major concern in the poultry industry and chicken male reproductive organs are the infectious tissues of various pathogenic microorganisms. Protection of the chicken male reproductive organs from pathogens is therefore an essential aspect of reproductive physiology. Recently Toll-like receptors (TLRs) have been identified as one of the key components of innate immunity in vertebrate species and have been reported to be expressed in the reproductive organs in various female species, including the chicken. However, mechanisms of antimicrobial protection of male reproductive organs mediated by TLRs are poorly understood. The objectives of this study were to determine the expression profile of the entire family of the ten chicken TLR genes in the chicken testis, to investigate whether sexual maturation affects their testicular mRNA abundance and to determine the changes in their expression levels in response to Salmonella enteritidis (SE) infection. RNA was extracted from the testis of healthy pre-pubertal, sexually mature and aged birds, and from sexually mature SE infected birds. RT-PCR analysis revealed that all TLRs, apart from TLR1-1 (TLR6), were expressed in the chicken testis. Quantitative real-time PCR analysis revealed that the testicular mRNA abundance of certain TLRs was developmentally regulated with respect to sexual maturation, while SE infection resulted in a significant induction of TLR2-1, 4, 5, 15 and 21 in the testis of sexually mature birds compared, to healthy birds of the same age. These findings provide strong evidence to suggest a key role of TLRs in the innate immune responses of chicken testis against Salmonella colonization.

High prevalence of prolonged QT interval duration in male patients with Cushing's disease.

UNLABELLED: Hypogonadal males have recently been shown to present prolonged QT interval, an electrocardiographic measure indicative of risk for fatal cardiac arrhythmias. Excess cortisol secretion induces low testosterone levels in male patients with Cushing's disease but no study has yet evaluated if this is accompanied by changes in QT interval duration. We therefore decided to evaluate whether male patients with Cushing's disease present changes in QT interval duration. QT interval was measured in electrocardiographic readings from 19 men and 35 women with Cushing's disease and age- and sex-matched controls were used for comparison. QT interval was corrected for heart rate according to Bazett's formula (QTc) and QTc >440 msec and >460 msec were taken as indicative of increased risk for torsade de pointes in men and women, respectively. Mean QTc was significantly longer in male patients compared with healthy controls (426.9±9.27 vs. 389.7±8.31, p<0.05) and 5 men with Cushing's disease presented prolonged QTc (prevalence 26%). By comparison, none of the women with Cushing's disease presented prolonged QTc. Hypokalemia and low testosterone appeared associated with long QTc. CONCLUSIONS: Male patients with Cushing's disease present prolongation of QT interval which may lead to measurements associated with high risk for ventricular arrhythmias. Both low testosterone levels and hypokalemia appear to contribute to long QT in men with Cushing's disease.

Effect of polymorphisms at the STAT5A and FGF2 gene loci on reproduction, milk yield and lameness of Holstein cows.

The objective of this study was to investigate the association of polymorphisms at the STAT5A and FGF2 gene loci with fertility, lactation milk yield and lameness in dairy cattle. Five hundred and eighteen primiparous Holstein cows were included in the study. Several reproductive traits were considered including conception rate (0/1) at first insemination, conception rate (0/1) in a 305-day lactation, number of inseminations per conception, interval (days) from calving to conception for cows that conceived in the first 305 days of lactation and age at first calving. Milk yield and lameness incidence were also recorded. Genotyping was performed using PCR-RFLP. The effect of allele substitution at each gene locus on reproductive traits, milk yield and lameness was assessed with single-trait mixed linear models. No significant associations were found between reproduction traits and any of the studied polymorphisms, apart from age at first calving, for which STAT5A polymorphism had a suggestive effect (P = 0.077). In addition, no significant effect of any polymorphism on lameness was found. Replacement of the C by G allele at the STAT5A locus was associated with a significant (P<0.05) increase in lactation milk yield, suggesting that this locus could be considered in gene assisted selection for the genetic improvement of milk production.

Effects of sexual maturation and Salmonella infection on the expression of Toll-like receptors in the chicken vagina.

Toll-like receptors (TLRs) are a critical component of the innate immune response in many vertebrates, including avian species. The recent findings of chicken TLRs (cTLRs) expression in ovarian follicles and in the chicken ovary in vivo, as well as the changes in their expression in response to lipopolysaccharide or Salmonella enteritidis (SE) infection, have broad implications for reproductive physiology and for the prevention of transmission of zoonotic diseases to humans through the consumption of contaminated poultry eggs. Because the main route of egg contamination is from infection of the oviduct and mainly from the vagina, the aim of this study was to investigate the expression of the ten cTLRs identified to date in the chicken oviduct in vivo, to determine whether sexual maturation affects their mRNA abundance and to investigate whether SE infection alters the expression of TLRs in the chicken vagina. RNA was extracted from the vagina of healthy prepubertal, sexually mature and aged birds, and from sexually mature and aged SE infected birds. RT-PCR analysis revealed that all types of cTLRs apart from TLR1-1 were expressed in the vagina of sexually mature birds. Quantitative real-time PCR analysis revealed that the mRNA abundance of TLR2-1, 2-2 and 4 differ with respect to sexual maturation in the chicken vagina. SE infection resulted in a significant induction of TLR5 and 15 in the vagina of sexually mature birds, and in a significant induction of TLR2-1, 4 and 15 in the vagina of aged birds, while a significant down-regulation was observed for TLR7 in the vagina of sexually mature birds. These findings suggest that a TLR mediated immune response mechanism exists in the chicken vagina, playing a crucial role in preventing microbial pathogens from being incorporated into newly forming eggs.

Threshold estimation based on a p-value framework in dose-response and regression settings.

We use p-values to identify the threshold level at which a regression function leaves its baseline value, a problem motivated by applications in toxicological and pharmacological dose-response studies and environmental statistics. We study the problem in two sampling settings: one where multiple responses can be obtained at a number of different covariate levels, and the other the standard regression setting involving limited number of response values at each covariate. Our procedure involves testing the hypothesis that the regression function is at its baseline at each covariate value and then computing the potentially approximate p-value of the test. An estimate of the threshold is obtained by fitting a piecewise constant function with a single jump discontinuity, known as a stump, to these observed p-values, as they behave in markedly different ways on the two sides of the threshold. The estimate is shown to be consistent and its finite sample properties are studied through simulations. Our approach is computationally simple and extends to the estimation of the baseline value of the regression function, heteroscedastic errors and to time series. It is illustrated on some real data applications.

Transcriptional profiling of Toll-like receptors in chicken embryos and in the ovary during sexual maturation and in response to Salmonella enteritidis infection.

One of the key members of the innate immune system in many vertebrate species is the family of Toll-like receptors (TLRs). These molecules, which initiate the innate immune response and mount an anti-microbial response in both vertebrates and invertebrates, have recently been identified in the chicken genome. The recent findings of chicken TLRs (cTLRs) expression in ovarian follicles during follicular growth and in response to lipopolysaccharide (LPS) infection are very important for reproductive physiology due to the transovarian transmission of Salmonella enteritidis (SE) in laying hens. The aim of this study was to investigate the expression of the ten cTLRs identified to date, in the chicken ovary in vivo and embryos during early embryonic development, to investigate whether sexual maturation affects their ovarian mRNA abundance and to investigate the transcriptional changes of TLRs in the chicken ovary in response to SE infection. RNA was extracted from embryos from day 3 to day 10 of embryonic development as well as from the ovaries of healthy prepubertal, sexually mature and aged birds, and from sexually mature and aged SE infected birds. RT-PCR analysis revealed that all TLRs apart from TLRs 1-1 and 2-2 were expressed in the ovary of sexually mature chickens, while all TLRs apart from TLR1-1 were expressed in the chicken embryos during embryonic development. Quantitative real-time PCR analysis revealed that the ovarian mRNA abundance of TLRs differ with respect to sexual maturation. SE infection resulted in a significant induction of TLR4, and 15 in the ovary of sexual mature birds, and in a significant induction of TLR15 in the ovary of aged birds, while a significant down-regulation was observed for TLR3 in the ovary of aged birds. These findings suggest that a TLR-mediated immune response mechanism exists in the chicken ovary.

Identification of chicken lysozyme g2 and its expression in the intestine.

Lysozyme is an important component of the innate immune system, protecting the gastrointestinal tract from infection. The aim of the present study was to determine if lysozyme is expressed in the chicken ( Gallus gallus) intestine and to characterise the molecular forms expressed. Immunohistochemical staining localised lysozyme to epithelial cells of the villous epithelium along the length of the small intestine. There was no evidence for lysozyme expression in crypt epithelium and no evidence for Paneth cells. Immunoblots of chicken intestinal protein revealed three proteins: a 14-kDa band consistent with lysozyme c, and two additional bands of approximately 21 and 23 kDa, the latter consistent with lysozyme g. RT-PCR analyses confirmed that lysozyme c mRNA is expressed in 4-day, but not older chicken intestine and lysozyme g in 4- to 35-day chicken intestine. A novel chicken lysozyme g2 gene was identified by in silico analyses and mRNA for this lysozyme g2 was identified in the intestine from chickens of all ages. Chicken lysozyme g2 shows similarity with fish lysozyme g, including the absence of a signal peptide and cysteines involved in disulphide bond formation of the mammalian and bird lysozyme g proteins. Analyses using SecretomeP predict that chicken lysozyme g2 may be secreted by the non-classical secretory pathway. We conclude that lysozyme is expressed in the chicken small intestine by villous enterocytes. Lysozyme c, lysozyme g and g2 may fulfil complimentary roles in protecting the intestine.

The use of three-dimensional ultrasound for fetal gender determination in the first trimester.

The objective of this study was to examine the accuracy of fetal gender prediction at a routine first trimester scan using three-dimensional (3D) ultrasound. 200 women were recruited for this study and they agreed to have a transvaginal scan for their routine first trimester scan for fetal anatomy and nuchal thickness measurement. 3D volumes were obtained and stored. Two examiners independently reviewed all the volumes and recorded their diagnosis of fetal gender and measured the angle between the genital tubercle and the skin overlying the sacrum. After studying the 3D volumes both examiners recorded a diagnosis of male or female in 150 cases (81.5%). In 34 cases (18.5%) either both (n=21) or one of them (n=13) could not comment on fetal gender by studying the saved volume. From these 150 cases correct prediction of fetal gender by both examiners was achieved in 85.3% of cases. In 6.7% of cases both examiners predicted the wrong gender while for the rest 8% of cases each examiner assigned different gender to the fetus (k=0.84; standard error 0.045). Angle measurements performed from the saved 3D volumes were highly reproducible. Gestational age did not affect the accuracy of gender identification. This study demonstrates that 3D ultrasound can be an effective and fast way of identifying fetal gender in the first trimester. The advantages of 3D ultrasound stem from its ability to virtually reproduce all required views.

The influence of maternal hematocrit on placental development from the first to the second trimesters of pregnancy.

OBJECTIVES: To study the influence of maternal hematocrit (Ht) and hemoglobin (Hb) levels on placental size and growth in the first and mid-second trimesters of pregnancy. SUBJECTS/METHODS: This was a prospective study performed at the fetal medicine unit of a university hospital. One hundred and eighty-one women with a singleton pregnancy were recruited at 11-14 weeks' gestation. For each case three scans of the placenta were performed, the first at recruitment and the following two at 3-week intervals. The volume of the placenta was measured at each visit using a three-dimensional ultrasound scanner. The maternal Hb and Ht were measured within 2 weeks of the first scan. RESULTS: The placental growth during the second trimester was inversely related to the Ht levels (r = -0.29, P = 0.001). It was also related to the Hb level (r = -0.20, P = 0.021). An increase of 0.1 units of Ht was associated with 38% less growth of the placenta (95% confidence interval: 18-54% less growth). DISCUSSION: This study demonstrates the effects of maternal environment on placental growth. Our data suggest that the levels of Ht appear to affect the placental growth during the second trimester. Further studies on the factors that regulate placental growth are needed to elucidate the pathophysiology of these interactions and their effect on pregnancy outcome.