Efficient Gene Knockout and Knockdown Systems in Neospora caninum Enable Rapid Discovery and Functional Assessment of Novel Proteins

Neospora caninum is a parasite with veterinary relevance, inducing severe disease in dogs and reproductive disorders in ruminants, especially cattle, leading to major losses. The close phylogenetic relationship to Toxoplasma gondii and the lack of pathogenicity in humans drives an interest of the scientific community toward using N. caninum as a model to study the pathogenicity of T. gondii .

Current Understanding of the Pathogenesis of Porcine Circovirus 3

Circoviruses are closed, circular, single-stranded DNA viruses belonging to the family Circoviridae and the genus Circovirus. To date, at least four porcine circoviruses (PCVs) have been recognized, including PCV1 to PCV4, respectively. Similar to PCV2 pathogenesis, PCV3 has been reported worldwide with myriad clinical and pathological presentations such as reproductive disorders, respiratory diseases, diarrhea etc. Current understanding of PCV3 pathogenesis is very limited since the majority of studies were mostly field observations. Interpretation of the results from such studies is not always simple. Various confounding factors affect the clinical appearance and pathological changes of the infected pigs. Recently, several experimental PCV3 infection studies have been reported, providing a better understanding of its pathogenesis. In this review, we focused on novel findings regarding PCV3 pathogenesis from both field observation and experimental infection studies. Possible factors involved in the conflicting results among the experimental infection studies are also discussed. This review article provides important insight into the current knowledge on PCV3 pathogenesis which would aid in prioritizing research in order to fill the knowledge gaps.

Clinical characteristics of patients with reproductive dysfunction and benign ovarian neoplasms

The study reports on the analysis of the anamnestic data of2044 women with reproductive disorders who underwent operative interventions for tumors and tumorlike masses of the ovaries during the period of 1989-1999 in the Research Center of Obstetrics, Gynecology and Perinatology of the Russian Academy of Medical Sciences. A high incidence rate о f complicated anamnesis and the coexisting somatic and extragenital diseases was revealed. The data are presented concerning the status of menstrual and reproductive function of the patients examined.

Reproductive health of the fem ale athletes

Physicians, including gynecologists, who deal with sportswomen, do not always consider in their everyday practice the changes in female reproductive function caused by severe physical exercises. A t the same time, this problem is so important, that in 1992 American College of Sports Medicine introduced the term Female Athlete Triad, describing interrelationships between eating disorders, amenorrhea and osteoporosis. It is well known that menarche in athletes occur later than in sedentary women, 5-50% o f athletes are amenorrheic, anovulation takes place in 16%, luteal phase deficiency -in 4 2 % of female athletes. The high rate of ovarian insufficiency among athletes and theirprobable unfavorable sequelae require the problems in treatment andprevention of reproductive disorders in female athletes to be solved. This must be done within the modem standards of reproductive medicine.

Modern aspects of biological modeling of sexual development disorders

Введение. Изучение механизмов развития репродуктивных нарушений в условиях эксперимента на крысах является одним из важных направлений современной патофизиологии. Крыса имеет функционирующий эстральный цикл, трехнедельную гестацию и гемохориальный тип плацентации. На циклических изменениях в яичниках и эпителии влагалища крысы базируется биологическое моделирование эндокринной гинекологической патологии. В настоящее время интерес представляет разработка экспериментальной модели нарушения полового созревания в периоде детства. Нарушение полового созревания может приводить к различным патологическим изменениям в сфере репродуктивного здоровья в будущем, актуальность данной проблемы очевидна. Цель работы - изучение современных аспектов моделирования преждевременного полового созревания у девочек. Методика. Анализ современных отечественных и зарубежных работ, касающихся механизмов преждевременного полового созревания и исследований в области моделирования данной патологии в экспериментах на крысах. Результаты. Преждевременное половое созревание у девочек - нарушение, проявляющееся развитием одного или комплекса признаков половой зрелости до 7 летнего возраста. Детерминация полового развития связана с особенностью генетических и эпигенетических факторов. К последним традиционно относят характер питания, стресс как адаптационную реакцию, интегрированную с активацией гормонопоэза. Фактор питания связан с функционированием гормонов жировой ткани, включая лептин, грелин, эффектами инсулиноподобного фактора роста. В ряде экспериментальных исследований, связанных с воздействием факторов питания, стресса и световой дезадаптации на репродуктивную систему животного, доказано значимое влияние последней на нейромедиаторные системы мозга. Наименее изученными в механизме преждевременного полового созревания остаются вопросы нейроэндокринной регуляции гонадной оси системой KISS/KISS1R. Продолжение изучения ассоциации изменения профиля нейромедиаторов моноаминового ряда и динамики кисспептина в эксперименте на крысах способно расширить представление о механизмах половой дифференцировки мозга и транслировать полученные данные в клиническую практику, связанную с обследованием девочек с преждевременным половым созреванием. Заключение. В связи с малой распространенностью и ограниченностью представлений о патогенезе преждевременного полового созревания, данная проблема требует детального изучения. Необходимо дальнейшее изучение патогенетических основ данной патологии в условиях биологического моделирования на самках крыс раннего возраста. Introduction. Studying pathogenetic mechanisms responsible for development of reproductive disorders in rat models is an important direction of modern pathophysiology. The rat has a functioning estrous cycle, a three-week gestation, and a hemochorial placentation. Biological modeling of endocrine gynecological pathology is based on cyclic changes in the ovaries and in the epithelium of the rat vagina. Currently, the development of an experimental model of puberty disorders in childhood is of interest. Premature puberty can lead to various pathological changes in future reproductive health. The relevance of this problem is obvious. The aim of this work was to study modern aspects of modeling premature puberty in girls. Method. Modern domestic and foreign reviews on the mechanism of premature puberty and studies of modeling this pathology in experiments on rats were analyzed. Results. Premature puberty in girls is a disorder manifested by the development of one or all signs of puberty as early as before the age of 7 yrs. Sexual development is associated with characteristic roles of genetic and epigenetic factors. The latter traditionally include nutrition and stress as an adaptive reaction integrated with the activation of hormone synthesis. The nutrition factor is related with the functioning of adipose tissue hormones, including leptin, ghrelin, and the effects of insulin-like growth factor. A number of experimental studies on rats addressing effects of nutrition, stress, and light maladaptation on the reproductive system have demonstrated its significant effect on brain neurotransmitter systems. Regarding the mechanism of premature puberty, the least studied issue is the neuroendocrine regulation of the gonadal axis by the KISS/KISS1R system. Continuing study of the association between changes in the profile of monoamine neurotransmitters and the dynamics of kisspeptin in experiments on rats can expand understanding of sexual differentiation mechanisms in the brain. The obtained data can be translated into clinical practice for the management of premature puberty in girls. Conclusion. Due to the rare prevalence of premature puberty and insufficient data on its pathogenesis, this problem requires detailed study. It is necessary to further study the mechanism of this pathology by biological modeling on female rats at an early age.

Effect of Electroacupuncture on Reproductive Disorders and Insulin Resistance in a Murine Polycystic Ovary Syndrome Model

Polycystic ovarian syndrome (PCOS) is a common, complex, and heterogeneous endocrine and metabolic disorder. There is no standardized treatment, and it therefore requires individualized therapies according to the symptoms and pathogenesis of each patient. The present study aimed to determine the effect of electroacupuncture at the acupoints Zusanli (ST36), Sanyinjiao (SP6), and Neiguan (PC6) on reproductive disorders and insulin resistance in a murine model of PCOS induced by dehydroepiandrosterone (DHEA). Vaginal smear analysis was used to determine mice estrous cycle; intraperitoneal glucose and insulin tolerance tests were adopted to analyze metabolic characteristics; enzyme-linked immunosorbent assay was used to measure hormone levels; gene expression was quantified with real-time PCR; hematoxylin and eosin staining was used to observe ovarian morphology. We observed disordered estrous cycle, polycystic ovarian morphology, and higher levels of homeostasis model assessment-insulin resistance (HOMA-IR) and testosterone (T), indicating successful modeling of PCOS. DHEA increased levels of estrogen (E2), progesterone (P), testosterone (T), luteinizing hormone (LH), and follicle-stimulating hormone (FSH), and EA treatment restored them to levels seen in the control group. EA reduced the days in estrus caused by DHEA, improved the abnormal sex hormone receptor genes, and attenuated the DHEA-induced histomorphological changes in mouse ovaries. The average expressions of the androgen receptor (AR), estrogen receptor (ER), luteinizing hormone receptor (LHR), and follicle-stimulating hormone receptor (FSHR) genes in the ovary greatly increased after DHEA treatment and significantly decreased in the DHEA + EA group. After EA treatment, the cystic follicle (CF) number was reduced and corpora lutea (CL) increased in the DHEA + EA group compared to the DHEA group. EA improved glucose intolerance and insulin intolerance. Statistical analysis of intraperitoneal glucose tolerance test-area under curve (IPGTT-AUC) glucose levels revealed a significant decrease in DHEA group mice compared to the control and DHEA + EA groups. EA was found to restore fasting blood glucose, fasting serum insulin, and HOMA-IR. In summary, our study suggests that EA has a remarkable effect in the DHEA-induced murine PCOS model. Management of EA could improve estrous cycle, hormonal disorders, abnormal sex hormone receptors in ovaries, ovary morphology, and insulin resistance in PCOS mice.

Analysis of culling reasons during the breeding cycle and lifetime performance: The strategy to remove crossbred Landrace and Large White sows under tropical climate

Background and Aim: Sow culling is an important practice in commercial swine production because it is directly associated with the economic efficiency of the breeding herd. This study was conducted to analyze the reasons for sow culling and quantify the factors affecting culling in crossbred Landrace and Large White sows under tropical climate. Materials and Methods: A total of 4887 culled sows from one parent stock farm located in Ratchaburi province, Western Thailand, were examined in this study. Culling reasons were grouped into the following eight categories according to farm management: (1) Reproductive disorders, (2) old age, (3) low performance, (4) diseases, (5) lameness, (6) udder problems, (7) body condition, and (8) other illnesses. Logistic regression analysis was used to explore the relationship between culling sows and environmental factors. Effects of parity and season of culling were considered as fixed effects in a statistical model. Results: Descriptive statistics indicated the following factors accounting for sow removals: Old age (34.93%, n=1707), reproductive disorders (29.32%, n=1433), low performance (12.62%, n=617), lameness (12.56%, n=614), diseases (4.8%, n=235), body condition (4.68%, n=229), udder problems (0.79%, n=39), and other illnesses (0.26%, n=13). Parity and season of culling were also found to have a significant effect on sow culling (p<0.05). The majority of culling sows in this population were of old age and high parity. Conclusion: This study indicated that the purposeful culling of sows on this farm was within the targeted range. However, the incidence of reproductive disorders was too high and required further investigations.

Importance of herpes simplex virus in development of infertility in married couples

The article considers the role of herpes simplex viruses of 1st, 2nd types in the development of reproductive disorders in married couples. The clinical example shows the difficulties of diagnosis, possible errors in treatment approaches, and the lack of results from the use of artificial insemination methods in the presence of persistent herpetic infection. Prolonged antiviral therapy led to the onset of pregnancy and the birth of a child.

Risk factors for erectile dysfunction: known and unexpected facts (review)

The prevalence of erectile dysfunction (ED) among men over 50 years is 31-51 %. By 2025, the prevalence of ED is expected to increase to 322 million men worldwide. In Russia, according to academician D.Yu. Pushkar', ED suffers up to 89.9 %.In this regard, it seems very important to establish the causes of this condition. The purpose of this review is to assess the current state of the literature on the issue of risk factors leading to or associated with the development of ED.The presented works show that well-known factors play a role in the occurrence of ED: unfavorable environmental conditions, harmful lifestyle factors, chronic intoxication, obesity, genetic predisposition, deficiency of sex hormones and vitamins, diseases of the cardiovascular system, prostate and urethra, as well as drugs for their treatment. Also, new indicators in laboratory and instrumental studies that change with ED are considered.Among the well-known data, there are also some very original works devoted to this topic. The relationship between ED and reproductive disorders, psoriasis, periodontitis, human immunodeficiency virus infection and some drugs use is now being investigated, which is presented in this review.

Endometrial Cytokines in Women with Reproductive Disorders

The purpose of this research was to study changes in endometrial cytokine concentrations in women suffering from reproductive disorders with and without chronic endometritis (CE) to justify pathogenetic treatment. Methods and Results: The study included 100 women of reproductive age with reproductive disorders. Group 1 included 50 patients with reproductive disorders and CE; Group 2 included 50 patients with reproductive disorders and without CE. Later on, all patients were divided into the following subgroups: Sub1A (n=31), and Sub2A (n=16) with an isolated bacterial flora, Sub1B (n=19) and Sub2B (n=34) with the absence of bacterial flora. The control group consisted of 31 fertile women. Endometrial aspiration pipe biopsy was performed on days 4-9 of the menstrual cycle (middle proliferative phase) using a disposable intrauterine probe (Taizhou Kechuang Medical Apparatus Co., Ltd, China) followed by histological examination of endometrial tissue. Laboratory diagnostics for sexually transmitted infections (STIs) was performed using the bacterial culture method. For the diagnosis of viral infection (HPV, HSV, CMV), cervical samples were studied using PCR. If STIs were detected, the patients were excluded from further research. Ultrasound examination of the pelvic organs was performed using the Aloka-5500 device with a 7MHz vaginal probe in two-dimensional visualization mode. The concentration of cytokines (IL-1β, INF-γ, TNF-α, ILs-4,6,8,10) in the endometrium was determined using the Protein Contour test systems (Saint Petersburg) and Multiskan EX ELISA Analyzer (Germany). In both groups, reproductive disorders were accompanied by hypoprogesteronemia and relative hyperestrogenemia, significantly apparent in CE. We found a 3-fold increase in the level of tissue pro- and anti-inflammatory cytokines (IL-1β, IL-4,6,10, INF-γ), and a 4-fold increase in the level of TNF-α and IL-8 in Group 1, compared to the CG. In Group 2, we found a 1.4-fold increase in the levels of IL-1β and INF-γ, compared to the CG. In Sub 1a, the levels of IL-6 and IL-8 were significantly higher than in the control group. In Sub1A, the isolated bacterial flora caused a cytokine inflammatory response characterized by a significant increase in the concentration of INF-γ and TNF-α, compared to Sub2A and Sub2B (P<0.05). In Sub1A, we found a tendency towards a decrease in the tissue levels of IL-4 compared to Sub1B and Sub2B; the IL-10 level was significantly lower than in Sub2B (P=0.0009) Conclusion: The results obtained in the present study showed the peculiarities of changes in cytokines at the level of endometrial tissue both in chronic inflammation of the endometrium and in its absence in women with reproductive disorders. The severity of the immune response is significantly higher in patients with CE, with the most significant change in the role of IL-10.