Search Results (26)

Background/Objectives: Beeswax alcohol (BWA; Raydel^®) is a blend of six long-chain aliphatic alcohols extracted from honeybee wax and is well known for its diverse functionality and health benefits. Herein, the efficacy of a BWA dietary intervention for 20 weeks was assessed to ameliorate high-cholesterol diet (HCD)-induced dyslipidemia and adverse effects on the vital organs of adult zebrafish. Methods: Adult zebrafish were fed different high-cholesterol diets (HCDs; final concentration of 4%, w/w) supplemented with BWA (final concentrations of 0.1%, 0.5% and 1.0%, w/w) or CoQ₁₀ (final concentration of 1.0%). Following 20 weeks of supplementation, blood and different organs (liver, kidney, testes and ovaries) were collected, and biochemical, histological and immunohistochemical analyses were performed. Results: The results demonstrate a dose-dependent effect of BWA of mitigating HCD-induced mortality in zebrafish over the 20-week supplementation period, which was noticeably better than the effect exerted by coenzyme Q₁₀ (CoQ₁₀). Consistently, a dose-dependent effect of BWA consumption of curtailing HCD-induced total cholesterol (TC) and triglyceride (TG) levels and increasing high-density-lipoprotein cholesterol (HDL-C) levels was noticed. Compared with CoQ₁₀ (final concentration of 1.0%, w/w), BWA (final concentration of 1.0%, w/w) displayed a significantly better effect of mitigating HCD-induced dyslipidemia, as evidenced by 1.2-fold (p < 0.05) and 2.0-fold (p < 0.05) lower TC and TG levels and 2.4-fold (p < 0.01) higher HDL-C levels. The histological analysis revealed substantial prevention of fatty liver changes, reactive oxygen species (ROS) generation, cellular senescence and interleukin (IL)-6 production in the hepatic tissue of BWA zebrafish, which was significantly better than the effect exerted by CoQ₁₀. Consistently, compared with CoQ₁₀, significant 25% (p < 0.05) and 35% (p < 0.01) reductions in the HCD-induced elevated levels of the hepatic function biomarkers aspartate aminotransferase and alanine aminotransferase was observed in the BWA group. Likewise, BWA consumption efficiently ameliorated HCD-induced kidney, ovary and testis damage by inhibiting ROS generation, cellular senescence and lipid accumulation. Conclusion: Supplementation with BWA demonstrated higher therapeutic potential than that with CoQ₁₀ to prevent dyslipidemia and organ damage associated with long-term consumption of HCDs. Full article

Adiponectin is an important adipokine involved in glucose and lipid metabolism, but its secretion and potential role in regulating glucose utilization during ovarian development remains unclear. This study aims to investigate the mechanism and effects of follicle-stimulating hormones (FSHs) on adiponectin secretion and its following impact on glucose transport in the granulosa cells of rat ovaries. A range of experimental techniques were utilized to test our research, including immunoblotting, immunohistochemistry, immunofluorescence, ELISA, histological staining, real-time quantitative PCR, and transcriptome analysis. The immunohistochemistry results indicated that adiponectin was primarily located in the granulosa cells of rat ovaries. In primary granulosa cells cultured in vitro, both Western blot and immunofluorescence assays demonstrated that FSH significantly induced adiponectin secretion within 2 h of incubation, primarily via the PKA signaling pathway rather than the PI3K/AKT pathway. Concurrently, the addition of the AdipoR1/AdipoR2 dual agonist AdipoRon to the culture medium significantly stimulated the protein expression of GLUT1 in rat granulosa cells, resulting in enhanced glucose absorption. Consistent with these in vitro findings, rats injected with eCG (which shares structural and functional similarities with FSH) exhibited significantly increased adiponectin levels in both the ovaries and blood. Moreover, there was a notable elevation in mRNA and protein levels of AdipoRs and GLUTs following eCG administration. Transcriptomic analysis further revealed a positive correlation between the expression of the intraovarian adiponectin system and glucose transporter. The present study represents a novel investigation, demonstrating that FSH stimulates adiponectin secretion in ovarian granulosa cells through the PKA signaling pathway. This mechanism potentially influences glucose transport (GLUT1) and utilization within the ovaries. Full article

Infertility is considered a global health issue as it currently affects one in every six couples, with female factors reckoned to contribute to partly or solely 50% of all infertility cases. Over a thousand genes are predicted to be highly expressed in the female reproductive system and around 150 genes in the ovary. However, some of their functions in fertility remain to be elucidated. In this study, 13 ovary and/or oocyte-enriched genes (Ccdc58, D930020B18Rik, Elobl, Fbxw15, Oas1h, Nlrp2, Pramel34, Pramel47, Pkd1l2, Sting1, Tspan4, Tubal3, Zar1l) were individually knocked out by the CRISPR/Cas9 system. Mating tests showed that these 13 mutant mouse lines were capable of producing offspring. In addition, we observed the histology section of ovaries and performed in vitro fertilization in five mutant mouse lines. We found no significant anomalies in terms of ovarian development and fertilization ability. In this study, 13 different mutant mouse lines generated by CRISPR/Cas9 genome editing technology revealed that these 13 genes are individually not essential for female fertility in mice. Full article

The spotted scat (Scatophagus argus) is an important ornamental species with sexually biased ornamental values that favor males. Therefore, it makes sense to breed mono-male fingerlings as ornamental fish. The spotted scat has an XX/XY sex determination system; therefore, the first step in producing genetically all-male offspring should be the induction of a fertile sex-reversed XY pseudo-female, which would then be mated with a normal XY male to produce a YY super-male. However, the XY pseudo-female produced by estradiol (E₂) treatment failed due to ovarian malformations. Here, male to female sex reversal was induced in spotted scat through a combination of E₂ and an androgen inhibitor (trilostane, TR). Spotted scat fingerlings of approximately 2.8–3.5 cm were fed a diet containing both E₂ (300 µg/g) and TR (300 µg/g). The fish were treated for 90 days and then fed a normal diet until they reached one year of age, when they were sampled. Twenty-eight treated XY individuals were identified using sex-linked markers. According to their gonadal histological characteristics, these treated XY fish could be divided into three groups: males with testes (n = 21), intersex individuals with ovaries–testes (n = 3), and fully sex-reversed individuals with ovaries (n = 4). All treated XX fish (n = 8) developed into normal females. There were no obvious abnormalities in the ovaries of the XY sex-reversed fish compared to the treated XX fish. Serum 11-ketotestosterone (11-KT) levels were significantly lower in XY-reversed females and treated XX females than in XY males. Compared with XY male fish, the expression of female (42sp50, foxl2, figla, zar1, and zp2) and male (dmrt1, gsdf, amh and cyp11b2) biased genes was up- and down-regulated, respectively, in the gonads of XY-completely sex-reversed fish. Immunohistochemical results indicate that 42Sp50 was expressed in oocytes of XY-partially and completely sex-reversed fish, while strong Gsdf signals were mainly detectable in testicular somatic cells of XY-partially sex-reversed fish and XY male fish. XY females were successfully produced by the combined treatment of E₂ and TR in the spotted scat. Their fertility will be tested in the future. Full article

The aim of this study was to determine the effects of silver nanoparticles (AgNPs) on the morphology and enzymatic activity of butterfly splitfin (Ameca splendens). Individuals of both sexes, aged about five months, were exposed to AgNPs at concentrations of 0 (control group), 0.01, 0.1, and 1.0 mg/dm³ for 42 days. On the last day of the experiment, the fish were euthanized, subjected to standard histological processing (anterior intestine, liver, and gonads), and analysed for digestive enzyme activity in the anterior intestine and oxidative stress markers in the liver. Fish in the AgNP 0.01 and 0.1 groups had the lowest anterior intestinal fold and enterocyte height. However, there were no statistically significant changes in the digestive enzyme activity in the anterior intestine. Analysis of enzymatic activity in the liver showed an increase in superoxide dismutase activity in fish in the AgNP 0.1 group. Histological analyses showed that AgNPs inhibited meiotic divisions at prophase I in a non-linear manner in ovaries and testes. In the AgNP 0.1 and 1.0 groups, the area occupied by spermatocytes was lower compared to the other groups. These results indicate that exposure to AgNPs may lead to disturbances in morphology and enzymatic activity in the liver and intestine and may lead to disruption of reproduction in populations. Full article

Tissue engineering has emerged as an indispensable tool for the reconstruction of organ-specific environments. Organ-derived extracellular matrices (ECM) and, especially, decellularized tissues (DCL) are recognized as the most successful biomaterials in regenerative medicine, as DCL preserves the most essential organ-specific ECM properties such as composition alongside biomechanics characterized by stiffness and porosity. Expansion of the DCL technology to cancer biology research, drug development, and nanomedicine is pending refinement of the existing DCL protocols whose reproducibility remains sub-optimal varying from organ to organ. We introduce a facile decellularization protocol universally applicable to murine organs, including liver, lungs, spleen, kidneys, and ovaries, with demonstrated robustness, reproducibility, high purification from cell debris, and architecture preservation, as confirmed by the histological and SEM analysis. The biomechanical properties of as-produced DCL organs expressed in terms of the local and total stiffness were measured using our facile methodology and were found well preserved in comparison with the intact organs. To demonstrate the utility of the developed DCL model to cancer research, we engineered three-dimensional tissue constructs by recellularization representative decellularized organs and collagenous hydrogel with human breast cancer cells of pronounced mesenchymal (MDA-MB-231) or epithelial (SKBR-3) phenotypes. The biomechanical properties of the DCL organs were found pivotal to determining the cancer cell fate and progression. Our histological and scanning electron microscopy (SEM) study revealed that the larger the ECM mean pore size and the smaller the total stiffness (as in lung and ovary), the more proliferative and invasive the mesenchymal cells became. At the same time, the low local stiffness ECMs (ranged 2.8–3.6 kPa) did support the epithelial-like SKBR-3 cells’ viability (as in lung and spleen), while stiff ECMs did not. The total and local stiffness of the collagenous hydrogel was measured too low to sustain the proliferative potential of both cell lines. The observed cell proliferation patterns were easily interpretable in terms of the ECM biomechanical properties, such as binding sites, embedment facilities, and migration space. As such, our three-dimensional tissue engineering model is scalable and adaptable for pharmacological testing and cancer biology research of metastatic and primary tumors, including early metastatic colonization in native organ-specific ECM. Full article

Anthonomus grandis Boheman (Coleoptera: Curculionidae) survives on alternative diets; however, this induces reproductive diapause. The objective was to evaluate the morphology and morphometry of the reproductive tract of this weevil after feeding on alternative diets. The experimental design was completely randomized with 160 replications and treatments arranged in a factorial design 3 × 3, represented by A. grandis adults fed on 3 food types (fragments of banana (T1) or orange (T2) endocarp, or with cotton squares of cultivar BRS 286 (T3, control)) and three evaluation periods (30, 60, and 90 days) and after each of these periods they were fed with cotton squares for 10 days. The reproductive tract of 100% of A. grandis females fed banana endocarp, orange endocarp, and cotton squares for 30 and 60 days and then cotton squares were morphologically adequate for reproduction, and after 90 days, only 50% of those fed cotton squares were in this condition. The length of the ovarioles and the width of the mature oocyte were greater for A. grandis fed on cotton squares and smaller in those with banana and orange endocarps. Histological sections reveal that male testes even with strong degenerative signals are already producing spermatozoa. On the other hand, females displayed ovaries with nurse cells in the tropharium and some maturing oocytes in the vitellarium. The body length was longer and the testis area and diameter smaller in males fed on cotton squares than in those with banana and orange endocarp. Anthonomus grandis females fed for ≥90 days with alternative food sources do not recover the functionality of their reproductive tract, even after consuming, for 10 days, a diet that favors reproduction. On the other hand, the males remain with their reproductive organs functional with this condition. Full article

The steroid 17α-methyltestosterone (MT) inhibits ovarian function and is often used to induce sex reversal artificially in vertebrates. In the present study, different concentrations of MT were added as dietary supplementation, and the effects on sex ratio, growth, and gonadal development were examined. After 40 days, the sex ratio (male:female) in each group increased at different degrees with 50 (1.36:1), 100 (1.57:1), and 200 (2.61:1) mg/kg MT, and neo-males with testis–ovary coexistence were observed in the 200 mg/kg MT group. Furthermore, 50 and 100 mg/kg MT could induce female reversion in neo-males. Histologically, the development of the testes in experimental groups was slower, but the ovaries of the experimental and control groups had similar developmental rates. The expression levels of DMRT11E, Foxl2, and SoxE1 in males at 200 mg/kg MT were 8.65-, 3.75-, and 3.45-fold greater than those of the control group. In crustaceans, sex reversal through vertebrate sex hormones can be observed. Neo-males (sex-reversed female prawns) were maintained by exogenous androgen, and over-reliance led to slow testis growth, small body size, and low growth rate, but sperm was still produced. In female prawns, MT inhibited ovary development and promoted growth. Full article

Collision tumors (CT) consist of two independent neoplasms with distinct neoplastic populations. Disorders of sexual development (DSDs) are characterized by atypical sexual development leading to various abnormalities of the genital tract. Sex reversal (SR) syndromes are a type of DSD characterized by a discrepancy between chromosomal sex and gonadal development (testes/ovaries) and the presence or the absence of the SRY gene. A phenotypically female 8-year-old Jack Russell terrier dog was referred due to anomalous vaginal discharge and non-pruritic cutaneous bilateral symmetrical alopecia on the flanks. During abdominal palpation, a voluminous mass was detected in the left quadrant area, later confirmed by ultrasound. The owner decided to proceed with euthanasia and necropsy. In the abdominal cavity, the left gonad was increased in size, the right one and the uterus were decreased, and the vagina and vulva appeared to be thickened. Histologically, both gonads were revealed to be testes: the left one was affected by a double neoplastic component (sustentacular tumor and interstitial cell tumor), whereas the right gonad showed coarctated seminiferous tubules. PCR amplification of the genes SRY and AMELX revealed the absence of the MSY region of the Y chromosome. To the authors’ knowledge, this is the first report describing a case of a testicular collision tumor in a DSD SRY-negative dog. Full article

The Red Palm Weevil (RPW) is one of the major pests of palms, frequently leading to the plants death. Action plans and the development of bio/physical strategies to contrast RPW diffusions are strongly recommended due to the serious concerns related to environmental pollution and insects’ resistance to chemicals. In the present study, we investigated morphological alterations of the ovaries and testes in adult RPW exposed to 2.45 GHz for 5, 15, and 30 s. During these treatments, the relative increase in temperature and the days of survival after irradiation were monitored. Then, RPWs were processed for macroscopical and microscopical analysis. Histological lesions of the ovaries and testes were characterized by the degeneration and necrosis of germinal cells, which increased with the increase in the time of irradiation and the temperature. By the same token, an increase in the temperature of irradiated insects was associated with a decrease in their survival time. These observations lead us to conclude that MWs could represent a useful tool for reducing or eliminating the reproductive capacity of this dreaded insect. Full article

The present study was conducted to understand the changes in gonads and hematological parameters in climbing perch (Anabas testudineus) exposed to synthetic estrogen and progestogen [mixture of ethinylestradiol (EE2) and desogestrel (DES)]. Climbing perch were exposed to four different concentrations of EE2/DES mixtures, viz. 0 ng of EE2 and DES/L (T0), 3 ng EE2 and 15 ng DES/L (T3), 30 ng EE2 and 150 ng DES/L (T30), and 300 ng EE2 and 1500 ng DES/L (T300) for 60 days. On days 45 and 60, samples were taken to assess changes in somatic indexes, gonad histology, and hematological parameters. The gonadosomatic index (GSI) increased in both females and males with increasing concentrations of estrogen mixtures except for T30 females, which was the lowest among all\four treatments. The hepatosomatic index (HSI) was observed to be increased in males as estrogen content increased. However, compared to fish at T0, HSI in female individuals did not vary in T30 fish, where the value was the highest among all the treatments. On day 45, histological observations showed no feminization or intersexuality but several germ-cell deformities in the ovary (adhesion, degenerated oocyte wall, degenerated granulose layer, increased interfollicular space, atretic follicle, and cytoplasmic clumping) and testes (increased interstitial area, focal loss of spermatocyte, dilation of the lumen, breakage of tubular epithelium, and elongated seminiferous tubule) were observed in fish exposed to EE2 and DES. Fish reared at T30 had lower RBC count, hemoglobin (Hb), glucose, and hematocrit levels. On day 60, fish reared at T30 had the highest Hb content compared to fish raised in other treatment conditions. WBC was progressively higher with increasing EE2/DES concentrations. Significant erythrocyte cytoplasmic abnormalities and erythrocyte nuclear abnormalities were observed in fish exposed to higher EE2/DES concentrations. The present study provides insights into the adverse impacts of synthetic estrogens sourced from human contraceptive pills on fish physiology. Full article

This study aimed to isolate and freeze germ cells from the superior brown mealworm. Styrofoam diet changes were observed for 20 days to determine whether mealworms were useful insects for decomposing Styrofoam. The average weight of mealworms before the Styrofoam diet was 500 mg, which decreased to 336 mg at D20 after their diet. To preserve mealworms with excellent Styrofoam-degrading ability, we first isolated the reproductive organs of mealworms, testes, ovaries, sperms, and ovarioles. Morphologically, male and female adult brown mealworms were distinguished according to the presence or absence of a protrusion at the tip of the fifth segment of the abdomen. Sperms and ovarioles were observed in anatomically isolated testes and ovaries. We compared mechanical and enzymatic (collagenase I) methods to effectively isolate ovarioles from adult female brown mealworms. For the enzymatic method, most were torn and burst as the membrane of the ovarioles was damaged by collagenase I, unlike the mechanical method. To preserve the superior genetic resources of mealworms, we cryopreserved the ovaries of female brown mealworms using slow-freezing and vitrification. Histological analysis showed that the yolk sac was completely damaged in the ovaries after slow-freezing. However, only partial damage was achieved in the vitrification group compared to the control group (no freezing). The newly developed vitrification method with alginate-encapsulated ovarioles maintained the yolk sac in the ovarioles but was evenly distributed. These results provide basic data for reproductive studies of other useful insects and contribute to the biobanking and fertility preservation of superior mealworm germ cells and endangered insects. Full article

The Sertoli cell tumor of the ovary is a rare ovarian tumor with non-specific symptoms. According to the literature, endocrine manifestations occur in two-thirds of patients, but testosterone production is extremely rare. Typically, it is a unilateral benign tumor of the ovary that most commonly presents in adolescents and young women of childbearing potential. We report a 29-year-old patient, previously diagnosed to have polycystic ovarian syndrome, who presented with complaints of amenorrhea for the past three years. A transvaginal ultrasound scan revealed polycystic structure ovaries and a solid cystic formation of 32 × 31 mm size with strong blood flow in the left ovary. The laboratory tests reported an elevated testosterone level. During laparoscopic surgery, a solid, yellowish tumor was removed and the left ovary was resected. Histological examination revealed a left ovary Sertoli cell tumor with an immature prepubertal-like Sertoli cell component. Following surgery, the serum testosterone levels returned to normal and the menstrual cycle became regular. Due to the substantially low incidence of ovarian Sertoli cell tumors, information on their clinical behavior, morphologic spectrum, optimal management, and prognosis is limited. They are characterized by a wide variety of clinical manifestations, treated surgically, and, if diagnosed at an early stage, have good prognosis. We emphasize the extraordinarily rare clinical presentation of this case report. Full article

Silver nanoparticles (AgNPs) are found in open waters, but the effect of their low concentrations on an organism’s homeostasis is not fully understood. The aim of the study was to determine the short-term exposure effects of AgNPs coated by PvP (polyvinylpyrrolidone) on the homeostasis of livers and gonads in zebrafish. Sexually mature zebrafish were exposed for seven days to silver ions (0.01 mg/dm³) or AgNPs (0.01; 0.05; 0.1; 0.5; 1.0 mg/dm³). On the last day, the liver, testes, and ovaries were subjected to a histology analysis. In the liver, we analyzed the expression of the cat, gpx1a, gsr, sod1, and cyp1a genes. On the last day of the experiment, the lowest survival rate was found in the AgNPs 0.05 mg/dm³ group. The histological analysis showed that AgNPs and silver ions cause an increase in the area of hepatocytes. The highest proliferation index of hepatocytes was found in the AgNP 0.05 mg/dm³ group. Furthermore, AgNPs were found to interfere with spermatogenesis and oogonesis as well as reduce the expression levels of the cat, gpx1a, and sod1 genes in the liver compared with the control group. Based on the results, it can be concluded that exposure to AgNPs causes cytotoxic changes in zebrafish, activates the immune system, negatively affects the process of meiosis in the gonads, and generates oxidative stress. Full article

Cereals are often contaminated with fumonisins, which are the toxic byproducts of mold. The aim of the study was to determine the effect of maternal exposure to fumonisins on the development and the liver function of the offspring at weaning. Two doses of fumonisins (60 and 90 mg/kg b.w.) were tested. The changes in the basal blood morphology, the biochemical parameters, the absolute and relative weights of the vital organs, and the changes in the cardiac and biceps brachii muscle histology were studied. The liver damage was assessed by evaluating the liver morphology and the common clinical liver panel. Maternal fumonisin intoxication caused a decrease in the body weight at birth and an increase in the heart, liver, kidney, lungs, ovaries, and testes weights. The cytokines and hormones, as well as the red blood cell counts and hemoglobin levels, were elevated in a dose-dependent manner following the exposure to fumonisins. Maternal exposure caused degenerative morphological and structural changes in the liver, as well as inflammation in the striated muscles, such as the heart and biceps brachii, and disproportionate development of the rat offspring in a dose-dependent manner. Moreover, FB exposure resulted in the disproportional development of the rat offspring in a dose-dependent manner, which was probably caused by the bodily hormonal dysregulation. Prenatal fumonisin exposure can be a pathological precursor for serious diseases, such as obesity and diabetes, later in life. Full article