Male Animals Research Articles

Sevoflurane inhibited reproductive function in male mice by reducing oxidative phosphorylation through inducing iron deficiency.

Sevoflurane (Sev) is one of the commonly used inhalation anesthetic chemicals in clinics. It has great impact on spermatogenesis and fertilization in male animals. The underlying mechanism remains largely unexplored. Based on our previous research, we hypothesized that Sev induced iron metabolism disturbance in the testis and epididymis and inhibited the spermatogenesis. In this study, two-month-old C57BL/6 male mice were treated with 3% Sev for 6h, and their fertility (including sperm concentration, sperm mobility, and the number of offspring) was evaluated. Mice testis, epididymis, and sperm were harvested and subjected to Western blot analysis and immunofluorescence analysis. Iron levels were reflected by the gene expression of iron metabolism-related proteins (including ferritin, TfR1, and FpN1) and ICP-MS and Perl's iron staining. Electron transport and oxidative phosphorylation levels were measured by Oxygraph-2k and ATP contents. The activity of ribonucleotide reductase was evaluated by assay kit. DNA synthesis status in testis and/or epididymis was marked with BrdU. Cell proliferation was evaluated by double immunofluorescence staining of specific protein marker expression. Our results revealed that the mice exposed to Sev showed damaged testicular and epididymis structure and significantly reduced the sperm concentration, sperm motility, and fertility. Sev decreases the iron levels through down-regulating the expression of H-ferritin, L-ferritin, and FpN1, and up-regulating the expression of TfR1 in the testis and epididymis. Iron levels also significantly reduced in germ cells which decrease the number of germ cells, including sperm, Sertoli cells, and primary spermatocyte. Iron deficiency not only decreases electron transport, oxidative phosphorylation level, and ATP production but also suppresses the activity of ribonucleotide reductase and the expression of Ki67, DDX4, GATA1, and SCP3, indicating that Sev affects the spermatogenesis and development. Meanwhile, Sev impaired the blood-testis barrier by decreasing the ZO1 expression in the testis and epididymis. The damage effect induced by Sev can be significantly ameliorated by iron supplementation. In conclusion, our study illustrates a new mechanism by which Sev inhibits spermatogenesis and fertility through an oxidative phosphorylation pathway due to iron deficiency of epididymis and testis or sperm. Furthermore, the damaging effects could be ameliorated by iron supplementation.

Seroprevalence and risk factors of bovine fasciolosis in the municipalities of Colombia.

Bovine fasciolosis is a reemerging neglected disease with a worldwide distribution caused by the trematode Fasciola spp., which parasitize various hosts. Bovine fasciolosis is responsible for large economic losses in the bovine livestock sector. This study aimed to estimate the seroprevalence and risk factors of bovine fasciolosis in the municipalities of Colombia. This was a descriptive cross-sectional study with simple random sampling conducted on 1140 cattle from the municipalities of Chiquinquirá, San Miguel de Sema, and Ubaté for a duration of 3 months. Serum samples were processed using the commercial Fasciola hepatica Antibody Test Kit IDEXX® Fasciolosis Verification (IDEXX, United States), which identified immunoglobulin G antibodies for gf2 antigen purified from Fasciola extracts. The f2 antigen is extremely immunogenic and highly specific for F. hepatica. An epidemiological survey was performed to record variables related to the sampled animals and herd management practices. Data were processed using the statistical program Epi Info® (Centers for Disease Control and Prevention; Atlanta, Georgia). The prevalence ratio was estimated to evaluate the association between fasciolosis and the hypothesized causal factors and the significance of this association using Pearson's Chi-square test. Finally, a logistic regression model was developed. The overall seroprevalence was 72.3%. The seroprevalence was 83.9% (323/385) in Chiquinquirá, 68.17% (257/377) in Ubaté, and 64.55% (244/378) in San Miguel de Sema. The seroprevalence was higher in male animals in Chiquinquirá and in female animals in San Miguel de Sema and Ubaté. Similarly, sex showed a statistically significant association with disease prevalence in Ubaté. The highest prevalence was found in cattle aged >2 years. The Holstein breed showed maximum seroprevalence in Chiquinquirá (p ≤ 0.05) and San Miguel de Sema, whereas crossbreed showed higher seroprevalence in Ubaté. Similarly, in Chiquinquirá, the association between the seroprevalence of fasciolosis and the presence of other species was statistically significant (95% confidence interval [CI]: 0.9601-3.4944; p = 0.0448). In Ubaté, the disease presentation was also associated with pasture rental (95% CI: 0.4047-1.0023; p = 0.003) and attendance to livestock expositions (95% CI: 0.2313-1.0636; p = 0.044). However, in San Miguel de Sema, water from the stream showed a statistically significant association with disease presentation (95% CI: 0.5209-1.0985; p = 0.00649785). Female sex and diarrhea occurrence were considered risk factors for fasciolosis. A high seroprevalence of antibodies to Fasciola spp. was detected in cattle in the study municipalities, indicating a high parasite distribution in these areas. Female sex and diarrhea were established as risk factors associated with fasciolosis in Ubaté and San Miguel de Sema, respectively. Further, research is necessary to establish prevention and control programs against parasitosis.

Alleviation of ischemic brain injury by exercise preconditioning is associated with modulation of autophagy and mitochondrial dynamics in cerebral cortex of female aged mice

Evidence from clinical studies and preclinical studies supports that exercise preconditioning can not only reduce the risk of stroke but also improve brain tissue and functional outcome after stroke. It has been demonstrated that autophagy and mitochondrial dynamics are involved in ischemic stroke. However, it is still unclear whether exercise preconditioning-induced neuroprotection against stroke is associated with modulation of autophagy and mitochondrial dynamics. Although age and sex interactively affect ischemic stroke risk, incidence, and outcome, studies based on young male animals are most often used to explore the role of exercise preconditioning in the prevention of ischemic stroke. In the current study, we examined whether exercise preconditioning could modulate autophagy and mitochondrial dynamics in a brain ischemia and reperfusion (I/R) model of female aged mice. The results showed that exercise preconditioning reduced infarct volume and improved neurological deficits. Additionally, increased levels of autophagy-related proteins LC3-II/LC3-I, LC3-II, p62, Atg7, and mitophagy-related proteins Bnip3L and Parkin, as well as increased levels of mitochondrial fusion modulator Mfn2 and mitochondrial fission modulator Drp1 in the ischemic cortex of female aged mice at 12 h after I/R were present. Our results could contribute to a better understanding of exercise preconditioning-induced neuroprotection against ischemic stroke for the elderly.

Sex and Age Effects on Healthy Gingival Transcriptomic Patterns

Many chronic inflammatory diseases demonstrate demographic associations such as sex, age, and race-ethnicity. Periodontitis has been found to be increased with age and in males. This study used nonhuman primates representing a human-like model for periodontitis and examined the gingival transcriptome stratified on sex and age. Thirty-six Macaca mulatta in 4 age groups—young (<3 y), adolescent (3–7 y), adult (12–15 y), and aged (>17 y)—with a healthy periodontium were used to characterize gene expression in healthy gingival tissues. Gene expression was compared to clinical measures of bleeding on probing (BOP) and probing pocket depth (PPD). The results demonstrated sex differences in number of up- and downregulated genes that increased with age. Female animals generally showed elevated expression of genes related to host immunoinflammatory responses, and males showed increased expression of tissue structural genes. Gene expression correlations with BOP and/or PPD showed minimal overlap between the sexes, while male animals demonstrated substantial overlap in genes that correlated with both BOP and PPD clinical features. A cluster analysis of genes significantly different between sexes showed a clear sex and age discrimination in the young and adolescent animals. In the older groups, the genes clustered predominately by sex, irrespective of age group. A pathway analysis identified that significant gene expression patterns were quite similar in adolescent and adult animals, while the young and aged samples were quite distinct. The results confirmed substantial sex related variations in gingival tissue biology that were affected by age and observed even in adolescent animals. This suggests that “programming” of the gingival tissues related to sex can occur rather early in life and presage variations in future risk for periodontitis.

Radiosensitivity of rhesus nonhuman primates: consideration of sex, supportive care, body weight, and age at time of exposure

ABSTRACT Background Animal models are vital for the development of radiation medical countermeasures for the prophylaxis or treatment of acute radiation syndrome and for the delayed effects of acute radiation exposure. Nonhuman primates (NHPs) play an important role in the regulatory approval of such agents by the United States Food and Drug Administration following the Animal Rule. Reliance on such animal models requires that such models are well characterized. Methods Data gathered from both male and female animals under the same conditions and gathered concurrently are limited; therefore, the authors compared and contrasted here the radiosensitivity of both male and female NHPs provided different levels of clinical support over a range of acute, total-body gamma irradiation, as well as the influence of age and body weight. Results Under matched experimental conditions, the authors observed only marginal, but clearly evident differences between acutely irradiated male and female NHPs relative to the measured response endpoints (rates of survival, blood cell changes, and cytokine fluctuations). These differences appeared to be accentuated by the level of exposure as well as by the nature of clinical support. Conclusion Additional studies with both sexes under various experimental conditions and different radiation qualities run concurrently are needed.

Distribution and length of muscle spindles and their 3D visualisation in the medial gastrocnemius of male and female rats.

The spatial distribution of the medial gastrocnemius muscle spindles of 10 male and 10 female rats was analysed under a light microscope, and for the first time, visualised using a 3D model of the muscle. Serial cross-sections of the medial gastrocnemius muscles were separated into 10 divisions along with the proximo-distal axis. The muscle spindles of the rat medial gastrocnemius were predominantly distributed on the proximo-medial divisions of the muscle. There were no sex-related differences in the distribution of the studied receptors. A single division contained an average of 2.71 receptors for animals of both sexes. Moreover, the calculated lengths of male and female rat muscle spindles were comparable, and average lengths did not significantly differ (3.30 ± 1.47 mm for male and 3.26 ± 1.32 mm for female rats). Therefore, the present results fill gaps in recent observations concerning similarities in muscle spindle numbers between male and female animals, despite significant differences in muscle mass and size.

Testosterone ameliorated the behavioural deficits of gonadectomised rats and counteracted free radicals in a dosage-dependent manner

Testosterone deficiency may induce behavioural changes in individuals. Oxidative stress resulting from a redox imbalance may be implicated in the initiation and progression of neurobehavioural disorders. However, whether exogenous testosterone intervention in male gonadectomised (GDX) rats ameliorates oxidative stress and plays a neuroprotective role remains unknown. Therefore, we examined this hypothesis by performing sham or gonadectomy surgeries on Sprague–Dawley rats with or without supplementation with different doses of testosterone propionate (TP). Open field and Morris water maze tests were performed, the serum and brain testosterone levels, and oxidative stress markers were analysed. GDX and lower TP doses (0.5 mg/kg) induced reduced exploratory and motor behaviours, but impaired spatial learning and memory compared to Sham rats. Administration of physiological TP levels (0.75–1.25 mg/kg) to the GDX rats restored the behaviour observed in the intact rats. However, higher TP doses (1.5–3.0 mg/kg) induced increased exploratory and motor behaviours but impaired spatial learning and memory. These behavioural impairments were accompanied by a marked decrease in levels of antioxidant enzymes (superoxide dismutase and catalase) and an increase in lipid peroxidation levels in the substantia nigra and hippocampus. These findings indicate that TP administration can alter behavioural performance and induce memory and learning impairment, which may result from changes in redox homeostasis in male GDX animals.

A Cannabinoid Receptor 1 Agonist Reduces Light-induced Phase Delays in Male But Not Female Mice.

Animals adapt to a changing environment by synchronizing their circadian rhythms to different stimuli, the strongest and most reliable being the daily light-dark cycle. Photic information reaches the central circadian pacemaker, the suprachiasmatic nucleus (SCN), which drives rhythms in physiology and behavior throughout the brain and body. The endocannabinoid system (ECS) is a neuromodulatory system that is present within the SCN, including the primary receptor, cannabinoid receptor 1 (CB1). Exogenous cannabinoids that target CB1 inhibit the phase-shifting effects of light in hamsters, mice, and rats. Furthermore, there is evidence in cultured microglial cells that cannabidiol (CBD), a constituent of Cannabis sativa, alters core circadian clock genes, while the CB1 agonist delta-9-tetrahydrocannabinol (THC) does not. The CB1 agonist studies were conducted using male animals only, but cannabinoids exhibit sex-dependent effects in various aspects of physiology and behavior. In addition, the effects of CBD on circadian behavioral rhythms have yet to be investigated. Therefore, we decided to test the effects of acute injections of CBD or the CB1 agonist CP 55,940 on light-induced phase delays in male and female C57BL/6J mice. Animals received a single injection at circadian time (CT) 15.5, followed by a 10-min light or dark (sham) pulse at CT 16. Running-wheel activity was monitored to determine activity levels and the behavioral phase shifts from different treatments. We observed a sex difference in the magnitude of phase delay size in response to CP 55,940 administration. Males had attenuated phase delays with increasing doses of CP 55,940, while females did not differ from control. Various doses of CBD had no effect on the phase-delaying effects of light in either sex. Our results show a sex difference in the gating of photic phase shifts by CB1 activation.

Reproductive and developmental toxicity assessments of LPM3480392, a novel mu opioid receptor biased agonist in rats

Opioids remain the most powerful analgesics for moderate to severe pain but their clinical use, misuse and abuse has been an alarming medical problem, especially for those users at child-bearing age. Mu-opioid receptor (MOR) biased agonists have been suggested as superior alternatives with better therapeutic ratios. We recently discovered and characterized a novel MOR biased agonist, LPM3480392, which demonstrates robust analgesic effect, favorable pharmacokinetic performance, and mild respiratory suppression in vivo. To understand the safety profile of LPM3480392 on the reproductive system and embryonic development, this study evaluated the effects of LPM3480392 on the fertility and early embryonic development, embryo-fetal development, and pre- and postnatal development in rats. Results showed that LPM3480392 had mild effects on parental male and female animals, accompanied by subtle early embryonic loss and delayed ossification of fetal development during organogenesis period. In addition, although minor effects were found on normal developmental milestones and behaviors in the pups, there was no evidence of malformed effect. In conclusion, these results suggest that LPM3480392 has a favorable safety profile with only minor effects on the reproductive and developmental outcomes in animals, which support the development of LPM3480392 as a novel analgesic.

Urolithiasis pada Kucing Snow di K and P Clinic Surabaya

Urolithiasis is a disorder in the form of crystals or stones in the urinary tract such as the urinary bladder that occurs in both male and female animals. The condition of the formation of calculi due to supersaturation in urine consisting of one or several types of minerals. A case of urolithiasis in a Persian cat named Snow, female, 40 months old, 3.76 Kg, solid white hair color, came to the K and P Clinic and a physical examination was performed by palpation of the bladder which was filled with urine but not distended. Complete blood count (CBC) examination using blood samples showed leukocytosis, lymphocytosis, monocytosis, and thrombocytosis. The results of blood chemistry tests using blood serum samples showed hypoglycemic conditions and an increase in blood urea nitrogen (BUN) values. Calcium phosphate crystals are found on microscopic examination of the urine. Urinalysis test showed abnormal values ​​for ascorbic acid, microalbumin, protein, calcium, leukocytes, creatinine, blood, and protein to creatinine ratio. The formation of calculi looks like a hyperechoic mass and the presence of an acoustic shadow can be seen on ultrasonography (USG) examination of the urinary bladder. The handler given is cystotomy surgery.&#x0D; Keywords: Urolithiasis, calcium phosphate, ultrasound.

A porcine bladder model of pre-clinical urodynamics demonstrates increased afferent nerve activity during filling.

Urodynamics are the accepted gold standard for the evaluation of multiple forms of voiding dysfunction. However, the tests are expensive, invasive, poorly reproducible, and often prone to artifacts. Therefore, there is a pressing need to develop next-generation urodynamics. The purpose of this study was to develop a novel ex vivo porcine bladder urodynamics model with afferent pelvic nerve signaling that can be used as a preclinical surrogate for bladder sensation. Porcine bladders including the ureters and vascular supply were harvested from local abattoirs using an established protocol in both male and female animals. Ex vivo bladder perfusion was performed using physiologic MOPS (3-(N-morpholino) propanesulfonic acid) buffer solution. The pelvic nerve adjacent to the bladder was grasped with micro-hook electrodes and electroneurogram (ENG) signals recorded at 20 kHz. Bladders were filled with saline at a nonphysiologic rate (100 mL/min) to a volume of 1 L using standard urodynamics equipment to simultaneously record intravesical pressure. ENG amplitude was calculated as the area under the curve for each minute, and ENG firing rate was calculated as number of spikes (above baseline threshold) per minute. At the conclusion of the experiment, representative nerve samples were removed and processed for nerve histology by a pathologist (hematoxylin and eosin and S100 stains). A total of 10 pig bladders were used, and nerve histology confirmed the presence of nerve in all adequately processed samples. Vesical pressure, ENG firing rate, and ENG amplitude all increased as a function of filling. During filling tertiles (low fill: min 1-3, med fill: min 4-6, and high fill: min 7-10), normalized pressures were 0.22 ± 0.04, 0.38 ± 0.05, and 0.72 ± 0.07 (cmH2O). Similarly, normalized ENG firing rates were 0.08 ± 0.03, 0.31 ± 0.06, and 0.43 ± 0.04 spikes/minute, respectively, and normalized nerve amplitudes were 0.11 ± 0.06, 0.39 ± 0.06, and 0.56 ± 0.14) μV, respectively. Strong relationships between average normalized pressure values and averaged normalized ENG firing rate (r2 = 0.66) and average normalized ENG amplitude (r2 = 0.8) were identified. The ex vivo perfused porcine bladder can be used as a preclinical model for the development of next-generation urodynamics technologies. Importantly, the model includes a reproducible method to measure afferent nerve activity that directly correlates with intravesical pressure during filling and could potentially be used as a surrogate measure of bladder sensation.

Sexual maturation in male bowhead whales (Balaena mysticetus) of the Bering-Chukchi-Beaufort Seas stock

Since the mid-1970s, study has focused on reproductive biology of female bowhead whales, while little has been described for males. This study evaluates testicular morphology (mass and length) and histology in relation to body length to determine the onset of male sexual maturity. Mean testis mass and mean testis length were highly correlated. Body length and mean testis mass were significantly correlated and an inflection of increased testicular mass occurred at approximately 12.5-13.0m suggesting the onset of puberty, and also indicated by histologic findings. Biological variability and the fact that few male animals have been examined within this critical length cohort do not allow determination of the onset of maturity with higher precision. Too few mature males have been landed in spring to make statistical comparisons of testes mass with autumn-landed animals within specific size cohorts. Two large (15.7m and 17.7m) males landed in spring had relatively small inactive testes and were diagnosed as pseudohermaphrodites; body length and mean testis length and seminiferous tubule diameter (STD) were not correlated with the other ‘normal’ whales. The smallest male confirmed as mature based on the presence of spermatocytes was 12.7m. The largest testes measured (combined mass 203kg) were from a whale landed in autumn. Mean STD for individual whales ranged from 33.3-170.9m and increased with mean testis weight and whale length. The STD is similar within a testis regardless of region evaluated, with minor variability. Some variation was noted for transverse sections within a cross section for some whales but no pattern was evident.

Generation and Characterization of the First Murine Model of Alzheimer's Disease with Mutated AβPP Inserted in a BALB/c Background (C.B6/J-APPswe).

Numerous mouse models of Alzheimer's disease (AD) are available, but all suffer from certain limitations, thus prompting further attempts. To date, no one model exists with amyloidopathy in a BALB/c strain. To generate and characterize the C.B6/J-APPswe mouse, a model of AD with a mutated human gene for the amyloid-β protein precursor (AβPP) inserted in a BALB/c background. We analyzed five groups at different ages (3, 6, 9, 12, and 16-18 months) of C.B6/J-APPswe and wild-type mice (50% males and 50% females) for the main hallmarks of AD by western blotting, amyloid-β (Aβ) ELISA, immunocytochemistry, electrophysiology, and behavioral tests. The C.B6/J-APPswe mouse displays early AβPP and Aβ production, late amyloid plaques formation, high level of Tau phosphorylation, synaptic deficits (reduced density and functional impairment due to a reduced post-synaptic responsiveness), neurodegeneration caused by apoptosis and necroptosis/necrosis, microgliosis, astrocytic abnormalities, and sex-related differences in explorative behavior, anxiety-like behavior, and spatial long-term and working memories. Social housing is feasible despite the intra-cage aggressiveness of male animals. C.B6/J-APPswe mice develop most of the distinctive features of AD and is a suitable model for the study of brain atrophy mechanisms and of the differences between males and females in the onset of cognitive/non-cognitive deficits.

Male larval experience of cues from adult rivals alters lifetime sperm investment patterns in a sperm heteromorphic moth, Ephestia kuehniella.

Male animals may adjust their resource allocations for reproduction and other fitness functions in response to cues from rivals. For instance, adult males increase their investment in sperm for a higher paternity share when they perceive sperm competition risk in their surroundings. In nature, both juveniles and adults may coexist spatially and temporally. Yet, it is not clear how juvenile males of different ages respond to cues from adult rivals and fine-tune their lifetime investment in sperm production and ejaculation in any insect. Here we used the Mediterranean flour moth, Ephestia kuehniella, which produces both fertile eupyrene and infertile apyrene sperm, to explore this question. We demonstrate that the late, but not early, instar larvae are sensitive to adult male cues. As a response, they produce more sperm before emergence and their resultant adults have shorter mating latency and ejaculate more sperm in the first few matings. When the juvenile stage produces more eupyrenes, the adult stops making these sperm, but regardless of the number of apyrenes produced during the juvenile stage, the adult continues to make them. These findings suggest that the number of spermatogonia for eupyrenes may be limited and that for apyrenes may be flexible. Our results show that the insect does not trade off survival, mating frequency, body size, or testis size for sperm production in response to adult males during the larval stage. Knowledge created in the present study offers insight into the stage-dependent sensitivity of juvenile males to cues from adult rivals and subsequent lifetime resource allocations.

Sex effects on gingival transcriptomic patterns during initiation, progression, and resolution of periodontitis.

The prevalence and severity of periodontitis demonstrates altered population distribution with age, sex, and race and ethnicity. While males exhibit greater frequency of disease, particularly with aging, the underlying basis for this observation remains obscure. This study used a nonhuman primate (Macaca mulatta) model of experimental ligature-induced periodontitis in adult animals to evaluate gingival transcriptomic differences stratified based upon sex of the animal. The 18 animals represented humans ages 40-80 years, with gingival tissue samples obtained at baseline, 0.5 months (initiation), 1 and 3 months (progression), and at 5 months that were 60 days after ligature removal for clinical disease resolution. Microarray analysis was used to quantify gene expression profiles in the gingival tissues. The results demonstrated clear gene expression differences in healthy (baseline) tissues between the sexes, with elevations in females associated with immune responses and elevation in males related to tissue structural genes. With disease initiation, fewer genes differed between the sexes, while these differences were significantly increased in progressing disease and resolution, particularly in male animals. Overexpressed biological processes showed tissue structural/functional genes at initiation, with host response pathways altered during disease progression. Resolution samples generally demonstrated biological processes of cellular metabolism that differed from baseline healthy samples. The transcriptomic findings support sex as a biological variable in periodontitis using a nonhuman primate model of experimental periodontitis.

Integrated multi-omics analysis reveals insights into Chinese forest musk deer (Moschus berezovskii) genome evolution and musk synthesis.

Among the artiodactyls, male animals belonging to the Family Moschidae have a unique tissue, the musk gland, with the capability of musk synthesis. However, the genetic basis of musk gland formation and musk production are still poorly understood. Here, musk gland tissues from two juvenile and three adult Chinese forest musk deer (Moschus berezovskii) were utilized to analyze genomic evolution events, evaluate mRNA profiles and investigate cell compositions. By performing genome reannotation and comparison with 11 ruminant genomes, three expanded gene families were identified in the Moschus berezovskii genome. Transcriptional analysis further indicated that the musk gland displayed a prostate-like mRNA expression pattern. Single-cell sequencing revealed that the musk gland is composed of seven distinguishable cell types. Among them, sebaceous gland cells and luminal epithelial cells play important roles in musk synthesis, while endothelial cells master the regulation of cell-to-cell communication. In conclusion, our study provides insights into musk gland formation and the musk-synthesizing process.

Gold nanoparticle intratesticular injections as a potential animal sterilization tool: Long-term reproductive and toxicological implications

This study aimed to evaluate the gold nanoparticles (AuNPs) animal sterilizing potential after intratesticular injections and long-term adverse reproductive and systemic effects. Adult male Wistar rats were divided into control and gold nanoparticle (AuNPs) groups. The rats received 200 µL of saline or AuNPs solution (16 µg/mL) on experimental days 1 and 7 (ED1 and ED7). After 150 days, the testicular blood flow was measured, and the rats were mated with females. After mating, male animals were euthanized for histological, cellular, and molecular evaluations. The female fertility indices and fetal development were also recorded. The results indicated increased blood flow in the testes of treated animals. Testes from treated rats had histological abnormalities, shorter seminiferous epithelia, and oxidative stress. Although the sperm concentration was lower in the AuNP-treated rats, there were no alterations in sperm morphology. Animals exposed to AuNPs had decreased male fertility indices, and their offspring had lighter and less efficient placentas. Additionally, the anogenital distance was longer in female fetuses. There were no changes in the histology of the kidney and liver, the lipid profile, and the serum levels of LH, testosterone, AST, ALT, ALP, albumin, and creatinine. The primary systemic effect was an increase in MDA levels in the liver and kidney, with only the liver experiencing an increase in CAT activity. In conclusion, AuNPs have a long-term impact on reproduction with very slight alterations in animal health. The development of reproductive biotechnologies that eliminate germ cells or treat local cancers can benefit from using AuNPs.

Detection of Gonadotropin Releasing Hormone (GnRH) and its analogues in equine and canine urine by high-resolution Data Independent Acquisition (DIA).

Gonadotropin releasing hormone (GnRH) and its synthetic analogues are considered banned substances by racing industries. GnRH is used as a pharmaceutical to regulate the female oestrous cycle but the hormone is also thought to increase the production of testosterone in male animals. Using liquid chromatography in conjunction with high resolution mass spectrometry (LC-HRMS) and Data Independent Acquisition (DIA), a method is presented for the detection of intact and truncated peptides of GnRH and its analogues down to the low picogram level in equine urine. The study of the catabolism of GnRH and analogues in plasma, combined with the analysis of urine from administration studies, reveals a common pattern of peptide catabolites that can be used to guide the design of mass spectrometry-based screens for this class of drugs. This culminated in the successful detection of the peptide in two out-of-competition canine urine samples.

Interrupting the blood-testis barrier with a flutamide-loaded nanostructured lipid carrier: A novel nonsurgical contraceptive approach for male animals

Flutamide is an antagonist of testosterone, an essential hormone in male reproduction. However, the use of flutamide as a contraceptive agent for nonsurgical castration in veterinary practice remains challenging due to its poor bioavailability. Here, the flutamide-loaded nanostructure lipid carrier (FLT-NLC) was synthesized, and its biological effects were demonstrated by an in vitro blood-testis barrier model. The flutamide was incorporated into the nanostructure lipid carrier by a homogenization method resulting in a high encapsulation efficiency (99.7 ± 0.04%). The FLT-NLC was negatively charged (−27.90 ± 0.10 mV), with a nano size (182.13 ± 0.47 nm) and narrow dispersity index (0.17 ± 0.01). An in vitro release study demonstrated a slower release profile of FLT-NLC when compared with flutamide solution (FLT). The FLT-NLC at doses up to 50 μM showed no significant cytotoxic effects against mouse Sertoli cells (TM4) or mouse fibroblast cells (NIH/3T3) (p > 0.05). An in vitro blood-testis barrier with FLT-NLC demonstrated remarkable lower transepithelial electrical resistance when compared with those lacking FLT-NLC (p < 0.01). Moreover, FLT-NLC significantly decreased the mRNA expression of blood-testis barrier proteins, CLDN11 and OCLN. In conclusion, we successfully synthesized FLT-NLC and confirmed its potential antifertility effects on in vitro blood-testis barrier, thus indicating its possible application as nonsurgical contraception for male animals.

Metabolism-based targeting of MYC via MPC-SOD2 axis-mediated oxidation promotes cellular differentiation in group 3 medulloblastoma

Group 3 medulloblastoma (G3 MB) carries the worst prognosis of all MB subgroups. MYC oncoprotein is elevated in G3 MB tumors; however, the mechanisms that support MYC abundance remain unclear. Using metabolic and mechanistic profiling, we pinpoint a role for mitochondrial metabolism in regulating MYC. Complex-I inhibition decreases MYC abundance in G3 MB, attenuates the expression of MYC-downstream targets, induces differentiation, and prolongs male animal survival. Mechanistically, complex-I inhibition increases inactivating acetylation of antioxidant enzyme SOD2 at K68 and K122, triggering the accumulation of mitochondrial reactive oxygen species that promotes MYC oxidation and degradation in a mitochondrial pyruvate carrier (MPC)-dependent manner. MPC inhibition blocks the acetylation of SOD2 and oxidation of MYC, restoring MYC abundance and self-renewal capacity in G3 MB cells following complex-I inhibition. Identification of this MPC-SOD2 signaling axis reveals a role for metabolism in regulating MYC protein abundance that has clinical implications for treating G3 MB.