Gonadal Research Articles

Comparative analysis of the cryopreservation protocols efficiency of goat sperm (review)

The aim of this work is to analyze scientific and practical data on protocols for cryopreservation of sperm of breeding goats and the effectiveness of their use. Numerous studies have established that seasonality is a characteristic reproductive function, with the greatest sexual activity observed in the autumn-winter period [1]. Goat sperm has the following species-specific features: sperm membranes contain a high level of unsaturated fatty acids, while the cytoplasm of cells, on the contrary, contains an insignificant amount of antioxidant compounds. In connection with these spermatozoa, especially strong lipid peroxidation induced by free radicals (hydroxyl radicals, UK peroxide), which cause structural damage to sperm membranes during freezing-thawing or storage in a cold state at 2-4 ° C [25, 26]. Taking these facts into account, an effective cryopreservation protocol, according to the results of numerous studies, is additional filtration of sperm for effective removal of accessory sex gland secretion, use of a diluent without egg yolk, cooling at 4-5 °C for 1.5-4 hours, packing in 0.25 - 0.5 ml straws. When using 0.5 ml straws, freezing is carried out in two stages: in liquid nitrogen vapor at a height of 4 cm for 5 minutes, then further immersion in liquid nitrogen. When using 0.25 ml straws, freezing is carried out in three stages: perforated glasses are placed 16 cm above the liquid nitrogen level for 2 minutes, then 4 cm for 3 minutes, after which they are completely immersed in liquid nitrogen for long-term storage.

Resveratrol mitigates heat stress-induced testicular injury in rats: enhancing male fertility via antioxidant, antiapoptotic, pro-proliferative, and anti-inflammatory mechanisms.

This study investigates the protective effects of resveratrol (RSV) against heat stress (HS)-induced testicular injury in rats. Climate change has exacerbated heat stress, particularly affecting male fertility by impairing testicular function and sexual behavior. A total of 32 rats were allocated into four experimental groups: control, RSV control, HS control, and RSV + HS. The HS groups were subjected to a 43°C water bath for 20 min to induce testicular hyperthermia, while the RSV + HS group received 20 mg/kg of RSV starting just before HS and continuing for eight weeks. Our findings reveal that HS significantly impairs male sexual behavior, evidenced by reduced mount and intromission numbers, and increased latencies. It also negatively affects the reproductive system, decreasing the weights of testes (Cohen's d = 1.8), epididymis, and accessory sex glands, and deteriorating sperm profile parameters such as motility (Cohen's d = 2.1), viability, and morphology. Furthermore, HS notably decreases reproductive performance in female rats, reducing litter size, live births, and conception rates. Biochemically, HS decreases activities of key antioxidant enzymes in the testes-glutathione peroxidase, superoxide dismutase, and catalase-while increasing lipid peroxidation, nitrite levels, and proinflammatory cytokines (IL-1β and TNF-α). It also reduces serum levels of reproductive hormones like testosterone (Cohen's d = 2.0) and 17β-estradiol.These results were affirmed with the histopathological evaluation and the immunohistochemistry staining (Ki-67, PCNA, Bax 5, and caspase-3 protein expression). Remarkably, RSV treatment mitigated these adverse effects, restoring both physiological and biochemical parameters toward normal levels (e.g., testicular weight Cohen's d = 1.6, sperm motility Cohen's d = 1.9, and testosterone levels Cohen's d = 1.7). This suggests that RSV's antioxidative, anti-inflammatory, antiapoptotic, and androgenic properties could effectively counteract the degenerative impacts of testicular hyperthermia. This highlights the potential of RSV as a therapeutic agent against climate change-induced fertility issues in males.

Toward At-Home and Wearable Monitoring of Female Hormones: Emerging Nanotechnologies and Clinical Prospects.

Steroid hormones, especially progesterone (P4), estradiol (E2), and testosterone (T), are key bioactive regulators in various female physiological processes, including growth and development, ovulation, and the reproductive cycle, as well as metabolism and mental health. As lipophilic molecules produced in sex glands, these steroid female hormones can be transported through blood vessels into various body fluids such as saliva, sweat, and urine. However, the ultralow concentration of steroid hormones down to picomolar (pM) level necessitates great demands for ultrasensitive but low-cost analytic tools to implement accurate, point-of-care or even continuous monitoring in a user-friendly fashion. This review focuses on the latest advances in materials and nanotechnologies to allow the rapid detection of female hormones at the pM level or below and the potentials in at-home and wearable hormone monitoring. We specifically summarize the optical and electrochemical strategies in this category, particularly those affording low cost and portable signal readout for at-home use. Furthermore, emerging flexible/wearable innovations are highlighted, which allow the continuous hormone cycle tracking in a noninvasive manner. The potential of these techniques is discussed to address the need for real-time acquisition of the hormone fluctuation, facilitating health monitoring at home. Lastly, we provide a comprehensive introduction to the prospects of female hormone monitoring in clinical diagnosis and treatment, from the perspective of gynecology and reproductive medicine clinicians.

Causes and clinical characteristics of women with primary amenorrhea and ovarian or gonadal disorders at a quaternary hospital.

To evaluate the causes and clinical characteristics of primary amenorrhea and ovarian or gonadal disorders in women. The medical records of all women evaluated for primary amenorrhea between January 2002 and December 2023 were retrospectively reviewed. Women with missing data were excluded from the study. Disorders of the ovary or gonads (hypergonadotropic hypogonadism) were defined as increased serum follicle-stimulating hormone (FSH) levels (>40 mIU/mL). Over a 21-year period, after excluding 1 woman with incomplete data, the study included 87 women who met the inclusion criteria. The median age at presentation was 18 years (interquartile range [IQR], 17-20). The median estradiol level was 5 pg/mL (IQR 5-10.4), and the median FSH level was 80 mIU/mL (IQR 63.1-94.9). The most common cause of primary amenorrhea was Turner syndrome (TS), followed by 46,XX and 46,XY gonadal dysgenesis. The median height was 147 cm (IQR, 140-158.7), and 50 women (57.5%) had karyotypic abnormalities, with TS being the most common abnormality. TS was identified as the most common cause of primary amenorrhea with ovarian or gonadal disorders.

Evaluation of ethephon induced oxidative stress to gonadal disorder and its amelioration by ethanolic extract of shoot of Bambusa balcooa Roxb. in albino rat

Evaluation of ethephon induced oxidative stress to gonadal disorder and its amelioration by ethanolic extract of shoot of Bambusa balcooa Roxb. in albino rat

Cohort profile: Multicenter Networks for Ideal Outcomes of Rare Pediatric Endocrine and Metabolic Diseases in Korea (OUTSPREAD study).

Rare endocrine diseases are complex conditions that require lifelong specialized care due to their chronic nature and associated long-term complications. In Korea, a lack of nationwide data on clinical practice and outcomes has limited progress in patient care. Therefore, the Multicenter Networks for Ideal Outcomes of Pediatric Rare Endocrine and Metabolic Disease (OUTSPREAD) study was initiated. This study involves 30 centers across Korea. The study aims to improve the long-term prognosis of Korean patients with rare endocrine diseases by collecting comprehensive clinical data, biospecimens, and patient-reported outcomes to identify complications and unmet needs in patient care. Patients with childhood-onset pituitary, adrenal, or gonadal disorders, such as craniopharyngioma, congenital adrenal hyperplasia (CAH), and Turner syndrome were prioritized. The planned enrollment is 1,300 patients during the first study phase (2022-2024). Clinical, biochemical, and imaging data from diagnosis, treatment, and follow-up during 1980-2023 were retrospectively reviewed. For patients who agreed to participate in the prospective cohort, clinical data and biospecimens will be prospectively collected to discover ideal biomarkers that predict the effectiveness of disease control measures and prognosis. Patient-reported outcomes, including quality of life and depression scales, will be evaluated to assess psychosocial outcomes. Additionally, a substudy on CAH patients will develop a steroid hormone profiling method using liquid chromatography-tandem mass spectrometry to improve diagnosis and monitoring of treatment outcomes. This study will address unmet clinical needs by discovering ideal biomarkers, introducing evidence-based treatment guidelines, and ultimately improving long-term outcomes in the areas of rare endocrine and metabolic diseases.

Diet influence on male sexual maturation through interplay between insulin signaling and juvenile hormone in insects.

In animals, sexual maturation coincides with the development of sexual behaviors and reproductive system. These developmental events are influenced by diet and governed by endocrine signals. Here, for the first time in insects, we explored functional links between nutrition and juvenile hormone (JH) in the male reproductive physiology through the insulin signaling pathway (ISP) acting as a transducer of nutritional signals. We turned to the male moth Agrotis ipsilon for which sexual maturation, including accessory sex glands (ASGs) development concomitantly with antennal lobes (ALs) maturation for female sex pheromone processing and display of sexual behavior, is known to be JH- and diet-dependent. Indeed, a diet rich in sugars with sodium was previously shown to accelerate sexual maturation, which was achieved from the third day of adult life. In this study, we demonstrated that such a diet raised i) the expression of JH signaling actors (Methoprene-tolerant, Taiman, and Krüppel homolog 1) in ALs and ASGs, ii) the biosynthesis and circulating levels of JH, and iii) the expression of both insulin receptor (InR) and insulin-like peptides (ILPs) in corpora allata (CAs) and brain respectively. Insulin injection raised JH biosynthesis following increased HMG-CoA reductase expression in CAs; opposite effects were induced in InR-deficient males. Thus, we highlighted that promoting effects of a diet composed of sugars with sodium on male sexual maturation results from an early induction of ISP causing an increase in JH biosynthesis followed by a potentiation of JH actions on the development of ASGs and ALs in A. ipsilon.

Understanding the Crucial Role of Seminal Plasma Exosomes in Bull Fertility: A Review.

Bull fertility is a multi-factorial trait and is affected by many factors, such as nutrition, genetics, and epigenetics. Superior quality male germplasm with high genetic merit helps to improve the livestock production trait. To achieve the target of livestock production, the availability of superior male germplasm is a great concern. In developing countries, there is a gap between the highly fertile frozen semen doses produced and the highly fertile frozen semen doses required. Improving the quality of existing low-fertile semen from high genetic merit bulls seems to play a promising role in filling this gap. Seminal exosomes are extracellular vesicles secreted by the epithelial cells of the testis, epididymis, and accessory sex glands such as the prostate gland. They contain a cargo of bioactive molecules such as proteins, nucleic acids and various metabolites. These molecules are transferred to the spermatozoa during its maturation and help in sperm capacitation, maturation, acrosome reaction, and fertilisation. Studies reveal that seminal exosomes help to improve the sperm functionality of low-quality sperm. Identification of the molecular profile of exosomes of bulls with proven fertility and their addition in an extender containing low-fertile semen may help to ameliorate the sperm quality of low-fertile semen, which may eventually aid in generating quantities of highly fertile ejaculates.

Tritrichomonas foetus is not located in the accessory glands, epididymis, or testicles of infected bulls, and post mortem changes influence recovery of organism.

To determine the presence of Tritrichomonas foetus in the accessory sex glands, epididymis, and testicular tissue of T foetus-positive bulls and to assess the impact of post mortem tissue acidity on quantitative reverse-transcription PCR results. 8 bulls were euthanized from June 12 through June 15, 2023, and samples from their accessory sex glands, testicles, and epididymis were collected for T foetus testing. Preputial smegma was obtained before and after euthanasia and tested for T foetus by quantitative reverse-transcription PCR. The pH of the penile and preputial epithelial surfaces was measured. Tritrichomonas foetus was not detected in samples from the testicle, epididymis, or epididymal semen. A post mortem decrease in the pH of the penis and prepuce correlated with a rise in cycle threshold values. Tritrichomonas foetus was not present in samples taken, suggesting its absence in these specific reproductive tissues. A decrease in the pH of the penis and prepuce over time post mortem correlated with an increase in cycle threshold values in the samples. This relationship indicates that changes in pH may impact the detectability or stability of genetic material in these tissues, possibly affecting diagnostic outcomes as time from death progresses. This research enhances the understanding of T foetus pathology in bulls and suggests that epididymal semen recovery could be a viable method for preserving genetic material from valuable bulls that test positive for T foetus. It also emphasizes the need for timely post mortem sampling to ensure accurate detection and management of T foetus in the cattle industry.

Impact of Cyclophosphamide on Testis and Sexual Glands of Adult Albino Rats and Probable Recovery After Drug Withdrawal: A Histological and Immunohistochemical study

Background: Cyclophosphamide is a chemotherapy and an immune-suppressive drug which is used for treatment of malignant and immune –related disease. The current work was conducted to evaluate the histological structure of the male gonad and sexual glands during treatment and after discontinuation of cyclophosphamide. Material and Methods: Thirty male albino- rats were classified into three groups. The first one was treated by normal saline, the second and third groups received cyclophosphamide 150 mg/kg intra peritoneal every two days for one week. The third group regarded as recovery group and divided into two subgroups with one- or two-months’ recovery period. At the accomplishment of the work, blood was drowning for hormonal analysis (testosterone) then the animals were sacrificed. The testis, prostate and seminal vesicle were removed and prepare for histological and immunohistochemical staining. Results: The cyclophosphamide treated group (GII) showed reduction of serum testosterone level, the testicular and sexual glands sections revealed atrophic and degenerative changes of their lining epithelium with reduction of their function and changes of their Ki67 expression which indicate abnormal proliferative pattern of the cells. However, the histopathological changes of treated group showed no improvement after one or even two months of drug withdrawal. Conclusion: The present study pointed out the risk of infertility and gonadal toxicity during cyclophosphamide treatment which may be permanent even after termination of the treatment.

Dietary supplementation with rosemary essential oil improves genital characteristics, semen parameters and testosterone concentration in Barki rams

BackgroundMale reproductive performance is an essential part of sheep production; therefore, the use of natural antioxidants to improve sperm quality and maintain reproductive performance in males is very important. Hence, oral administration of rosemary essential oil (REO) was investigated to improve the fertility rate, including the ultrasonographic testes, epididymal tail and genital glands, as well as semen parameters and testosterone concentration. Sixty animals were splitted into two groups, each with 30 rams; the rosemary group (C + REO) received 2 mg/kg/bw and the control group (C-REO). Ultrasound images and blood samples were collected at 15, 30 and 45 days of the REO treatment.ResultsThe testis and epididymal tail ultrasonographic assessments demonstrated a significant enhancement in the C + REO group compared to the C-REO group. However, the rams in the C + REO group showed significant improvements in the pampini-form plexus, seminal vesicle, Cowper's and prostate genital glands compared to the C-REO group. The data showed that the sperm cell concentration (× 109/ml) and individual motility (%) were significantly improved in the C + REO group. Furthermore, ejaculate volume (ml) in the C + REO group was significantly higher than that in the C-REO group. While the animals treated with REO did not improve live spermatozoa (%), it reduced the abnormalities of spermatozoa (%) compared to the C-REO group. Also, the C + REO group significantly increased the testosterone concentration more than the C-REO group.ConclusionIt can be concluded that supplementation with 2 mg/kg/bw REO improves genital characteristics, semen parameters and testosterone concentration in Barki rams.

Cancer mortality risk in the Urals Cohort of Exposed Population Offspring

The objective of the study is to analyze solid cancer mortality and estimate the risk value of solid cancer mortality in the offspring of parents irradiated in the Southern Urals, depending on the dose to the parents' gonads as well as to conduct a preliminary assessment of the mortality risk from cancer of individual locations depending on the gonadal dose. The issue of the transgenerational effects of the human gonad exposure is still very important. There exists conclusive evidence of the presence of such effects in experimental animals. However, there is no proof of the existence of these effects in humans despite a great number of research on this subject. International scientific community and international organizations, UNSCEAR and ICRP being among them, regard this issue as the one that has not been solved yet and requires further studies. Urals Cohort of offspring of the population exposed on the Techa River and on the territory of the East Ural radioactive trace was established in the Urals research Center for Radiation medicine in 2022. The key feature of the cohort is the exclusion of the offspring with postnatal exposure. The size of the cohort as of September 2023 is 31,154 persons. The number of person-years over the whole follow-up period from 1950 through 2020 is 1 226 380. Mean dose to the gonads of mothers of all the cohort members is 41 mGy, to those of the fathers' — 35 mGy, mean combined gonad dose is 76 mGy. Over the whole follow-up period 3,774 deaths from all causes including 284 deaths from solid cancers were registered in the cohort. The mean age of the cohort members at the end of the follow-up period was 42 years. The risk of death was analyzed using Poisson regression by the programs of the EPICURE statistical software package. Three models of dose dependence were tested: linear, quadratic, and linear-quadratic models of paternal gonadal dose, maternal gonadal dose, and total gonadal dose. Mortality risk analysis of all solid cancers in the offspring cohort showed no statistically significant effect with parental gonadal dose (we have obtained positive although statistically insignificant values of solid cancers mortality risk) which is consistent with the results of other studies in human populations. At the same time, the analysis for the first time obtained a statistically significant linear dependence of the solid cancer mortality risk in offspring with father's age over 45 years depending on the dose to the father's gonads. The excess relative risk of death was 8.09/Gy, (0.51-22.93), p < 0.05. Also, for the first time, evidence of dose dependence of the mortality risk of lung cancer in male offspring on maternal gonadal dose, paternal gonadal dose, and combined gonadal dose was obtained. The values of excess relative risk of lung cancer and 95% confidence intervals according to the linear model were 5.39/Gy (0.46; 15.56) from paternal gonad dose, 4.36/Gy (0.15; 13.48) from maternal gonad dose, and - 3.95/Gy (0.55; 12.14) from combined gonad dose. Point risk estimates of lung cancer at this stage are characterized by wide confidence intervals and require additional studies to assess the influence of possible effect modifiers, but with a high degree of probability indicate the presence of dose dependence of this effect. Increasing the follow-up period and attained age, will increase the number of cancers in the future and will reduce dose response uncertainties and provide more accurate estimates of the risk of death in the offspring cohort.

Seasonal heterochrony of reproductive development and gene expression in a polymorphic salamander.

Life cycle evolution includes ecological transitions and shifts in the timing of somatic and reproductive development (heterochrony). However, heterochronic changes can be tissue-specific, ultimately leading to the differential diversification of traits. Salamanders exhibit alternative life cycle polymorphisms involving either an aquatic to terrestrial metamorphosis (biphasic) or retention of aquatic larval traits into adulthood (paedomorphic). In this study, we used gene expression and histology to evaluate how life cycle evolution impacts temporal reproductive patterns in males of a polymorphic salamander. We found that heterochrony shifts the distribution of androgen signaling in the integument, which is correlated with significant differences in seasonal reproductive gland development and pheromone gene expression. In the testes, androgen receptor (ar) expression does not significantly vary between morphs or across seasons. We found significant differences in the onset of spermatogenesis, but by peak breeding season the testes were the same with respect to both histology and gene expression. This study provides an example of how seasonal heterochronic shifts in tissue-specific ar gene expression can disparately impact seasonal development and expression patterns across tissues, providing a potential mechanism for differential diversification of reproductive traits.

B-178 A Sensitive LC-MS/MS Assay for Total Testosterone Quantification

Abstract Background Total testosterone immunoassay is a commonly ordered test for the diagnosis of gonadal disorders. However, immunoassays suffer from low specificity and sensitivity, which leads to inaccuracy especially in the low concentration range (< 100 ng/dL). As a result, commercial testosterone immunoassays have difficulties to accurately measure total testosterone levels for women, children, and hypogonadal men. LC-MS/MS has emerged as the method of choice for the accurate quantitation of steroid hormones. We presented here a sensitive laboratory-developed total testosterone assay, which was validated on two LC-MS/MS instruments. Methods An internal standard containing [2,3,4-13C3]-testosterone was added to each sample. The spiked serum sample was loaded on a supported liquid extraction (SLE) cartridge and eluted using methylene chloride. The elute was dried down under nitrogen and reconstituted in sample reconstitution buffer. Chromatographic separation was achieved using a Restek Ultra C18, 50 x 2.1 mm, 3µm analytical column and a total run time of each sample was 5 minutes. The assay was validated on two triple quadrupole mass spectrometers, API 6500 and API 4500. Results were compared with those of a reference laboratory. Results The assay was linear in the range of 1 to 1200 ng/dL for total testosterone and the AMR was set at 2-1200 ng/dL. The interday and intraday imprecision were less than 8%. Split sample comparison to a reference laboratory demonstrated acceptable accuracy with the slope of 1.053 and correlation coefficient (R2) of 0.991. The limit of detection (LOD) was 0.2 ng/dL and the limit of quantitation (LOQ) was 0.5 ng/dL. Results obtained from two instruments agreed well with the slope of 0.996 and the R2 of 0.999. No carryover or interferences was observed for this assay. Conclusions A LC-MS/MS based method was developed and validated for measuring total testosterone levels for female and children. This sensitive and specific assay will improve the diagnosis of hypogonadism and monitoring of testosterone-suppressing treatment.

CORRELATION BETWEEN BMI AND GONADOTROPIN HORMONES IN THE FEMALE GENDER

Introduction: The pituitary gland affects growth and development. Its main function is in the activity of sexual/reproductive hormones and in the functioning of the glands (thyroid gland, adrenal glands and gonads). Gonadotropin hormones (gonadostimulins) are so named because they stimulate the function of the gonads - male and female sex glands (testicles and ovaries) in which sex cells - sperm and egg cells are created. Overweight is a state of metabolic disorders where excess body fat accumulates to the extent that it can negatively affect human health. Obesity as a BMI factor is related to the functioning of the thyroid gland with its hormones and the pituitary gland with gonadotropin hormones. Purpose of the study: This paper aims to present the correlation between BMI and gonadotropin hormones in reproductive and postmenopausal women. The aim of the paper will also consist in the status of gonadotropin hormones and the functional state of these hormones by comparing them with metabolic disorders such as BMI, obesity and the synthesis of follicle stimulating hormone and gonadotropin hormone. Material and method: A total of 20 patients will be included in the research, divided according to age group into two categories. The first group will include patients aged 10-40 years, the second group patients over 41 years old. Blood (serum) samples will be taken from patients for analyzing gonadotropin hormones: FSH, LH, PRL and thyroid hormones: TSH, FT4, FT3. They will be analyzed with the modern automatic fluorescent enzyme immunoassay method (Vidas-Biomerie). The BMI of the body will be measured with a metric formula where the weight of the patient (in kg) divided by the height of the patient (in m2) set to the square power will give us the measure of body index-BMI (%). SPSS – Software package version 20.0 Numerical series were analyzed using central tendency measures (average, median, minimum values, maximum values), as well as distribution measures (standard deviation); To determine the regularity of the frequency distribution of the examined variables, the Shapiro-Wilk W Test was used; Non-parametric tests for two independent parameters were used to test the significance and difference between some numerical parameters with irregular frequency distribution (Mann Whitney U test); To determine statistical significance, a two-way analysis was used with a significance level (means) of p<0.05. Results: The results of the research will give a real insight into the data of gonadotropic hormones FSH, LH, PRL and thyroid hormones TSH, FT4, FT3 of the female sex in the Tetova region. The results of the research will also provide information about the correlation of BMI and gonadotropin and thyroid hormones. Conclusion: From the analyzes performed on patients of different age groups we come to the conclusion that: • Gonadotropin hormonal disorders can lead to the phenomenon of metabolic diseases such as BMI. • These disorders can have consequences such as: polycystic ovary diseases in women, Hashimoto's genetic syndrome, Graves, infertility in women, etc.

Quality of life domains in patients with endocrine disorders: A study in the Al Jouf region of Saudi Arabia

Abstract Introduction: Data on the burden of endocrine disease in the Al Jouf region of Saudi Arabia are not available. This work aims to describe the burden of endocrinopathies in residents of the Al Jouf region of Saudi Arabia. Materials and Methods: Case files of endocrinopathy patients accessing care at a local district hospital located in the Al Jouf region of Saudi Arabia were extracted from the hospital database and appropriately analyzed. Results: Records of 582 adult patients consisting of 184 (31.6%) males and 398 (68.4%) females were analyzed. Endocrine disorders recorded included diabetes mellitus (DM), thyroid disorders, hypothalamo-pituitary disorders, adrenal disorders, and gonadal disorders. All endocrine disorders predicted low quality of life (QoL), with DM having the highest odds (physical health: OR = 5.21, 95% CI = 2.23–10.42, P = 0.001; mental health domain: OR = 6.01, 95% CI = 2.71–13.79, P = 0.001; social relation domain: OR =4.27, 95% CI = 2.81–11.91, P = 0.001; environmental health domain: OR =5.01, 95% CI = 2.70–11.97, P = 0.001). Conclusion: Endocrine disorders predicted low QoL, with DM having the highest odds across all four QoL domains.

Transcriptome analyses reveal key features of mouse seminal vesicle during aging

Despite the significant morphological changes that occur in the seminal vesicles with aging, the transcriptomic characteristics remain largely unexplored. To address this, we performed bulk RNA sequencing on seminal vesicle samples from mice aged 3, 13, and 21 months to uncover transcriptomic alterations. Our findings reveal that aged seminal vesicles display cystic dilatation, epithelial hypoplasia, disordered muscle layers, fibrosis, and reduced proliferation capability. A comparison between 3-month-old and 21-month-old mice indicated that leukocyte-mediated immunity and leukocyte migration were the most significantly upregulated biological processes among differentially expressed genes (DEGs). Notably, several DEGs associated with "leukocyte migration," such as Vcam1, Cxcl13, and Ccl8, exhibited an increasing trend in transcriptomic and protein expression at three different time points in the seminal vesicles of mice. Additionally, we identified multiple aging-associated DEGs, including P21 and Tnfrsf1b. Two genes (Cd209f and Ccl8) were consistently upregulated across all six regions of the male reproductive glands (testis, epididymis, and seminal vesicle) in the comparison of bulk RNA datasets from 3-month-old and 21-month-old mice. These analyses highlight an enhanced state of immune and inflammatory response in aged seminal vesicles. This study represents the first exploration of the overall transcriptome landscape of seminal vesicles in a murine model of natural aging, offering new insights into the mechanisms underlying aging-related seminal vesicle dysfunction.

Localization of β-Nerve Growth Factor in the Stallion Reproductive Tract.

β-Nerve growth factor (β-NGF) is a protein produced in the reproductive tract of camelids (camels, llamas, and alpacas) that has been identified as the ovulation inducing factor in seminal plasma. β-NGF from seminal plasma deposited into the reproductive tract of the female camelid acts systemically to stimulate the secretion of luteinizing hormone (LH) from the anterior pituitary, which in turn induces follicle maturation and ovulation. The objectives of the present study were to determine if β-NGF is present in the reproductive tract of the stallion and identify the specific site(s) of production. The hypotheses were that β-NGF would be present in the stallion reproductive tract and would primarily be localized in Sertoli cells of the testes and the prostate gland. Immunohistochemistry on paraffin-embedded paraformaldehyde-fixed tissues was performed using a rabbit polyclonal anti-β-NGF antibody on a total of six male equine reproductive tracts, including a one-day old colt, a one-year-old colt, and four adult stallion tracts. Strong immunostaining was observed in the efferent ducts of the testes and the epithelial cells of the prostate, seminal vesicles, bulbourethral glands, and ampullae. Weaker β-NGF staining was noted in Leydig cells, Sertoli cells, and spermatogonia within the testes and in epithelial cells of the epididymis. In conclusion, immunohistochemistry revealed that β-NGF is present in the stallion reproductive tract, and the protein is primarily present in the efferent ducts of the testes and in all accessory sex glands.

Aqueous-Deficient Dry Eye Increases Lipid Layer Thickness Measured by Dynamic Tear Interferometry.

To investigate the impact of aqueous deficiency on lipid layer thickness (LLT) measurement in dry eye disease before and after treatment of aqueous-deficient dry eye (ADDE) using dynamic tear interferometry. This prospective comparative study included 230 eyes from 230 patients with ADDE. The treatment group with punctal plug insertion was compared with the control group. The average LLT (LLTave) was measured with a LipiView II tear interferometer at baseline and after 1 month. Correlation analyses were performed with the indices of the dynamics of the tear film lipid layer (TFLL). Baseline LLTave values were 116.1 ± 26.7 and 112.3 ± 27.6 nm in the treatment group (n = 121) and the control group (n = 109), respectively. At 1 month, LLTave significantly decreased in the treatment group (difference -41.8 [95% confidence interval (CI) -47.2 to -36.3], P = 0.003). The values of tear meniscus height (TMH) at baseline and 1-month follow-up were negatively correlated with LLTave (both P < 0.001). Spreading time of TFLL to equilibrium and the deviation of TFLL were positively correlated with LLTave (all P < 0.001) and negatively correlated with TMH (P < 0.001 and 0.009). In multivariate analysis (adjusted R2 = 0.411, P < 0.001), LLTave was associated with TMH (B = -1.068; P < 0.001), adjusting for age, sex, and meibomian gland expressibility. In ADDE, the TFLL was measured to be thicker than the normal range of 60 to 99 nm and became thinner with treatment. LLT was affected by not only meibum secretion but also aqueous status due to changes in the dynamics of TFLL.

Ultra-high static magnetic fields altered the embryonic division and development in Caenorhabditis elegans via multipolar spindles

IntroductionUltra-high static magnetic fields (SMFs) have unique advantages in improving medical and academic research. However, the research on the early embryo exposure of ultra-high SMFs is minimal, extensive exploration is indispensable in living organisms. ObjectivesThe present study was aimed to study the effects of ultra-high SMFs on the early embryonic division and development of Caenorhabditis elegans (C. elegans). MethodsEarly adult parents containing fertilized eggs in vivo were exposed to SMFs at intensities ranging from 4 T to 27 T. The number of mitotic cells in the reproductive glands of the P0 worms, early embryonic cell spindle localization, embryo hatching and the reproductive as well as developmental indicators of F1 and F2 nematodes were examined as endpoints. ResultsOur results indicated that ultra-high SMFs has no obvious effect on the germ cell cycle, while 14 T and 27 T SMFs significantly increased the proportion of multi-polar spindle formation in early embryonic cells, and reduced the developmental rate and lifespan of C. elegans exposed at the embryonic stage. Spindle abnormalities of early embryonic cells, as well as the down-regulation of genes related to asymmetric embryonic division and the abnormal expression of the non-muscle myosin NMY-2 in the division grooves played a critical role in the slowing down of embryonic development induced by ultra-high SMFs. ConclusionsThis study provided novel information and a new sight for evaluating the biosafety assessment by exposure to ultra-high SMFs at the early embryonic stage in vivo.