Impact of reducing lipid content during in vitro embryo production: A systematic review and meta-analysis.

There is still no consensus regarding the role of lipid modulators during in vitro embryo production. Thus, we investigated how lipid reducers during the in vitro maturation of oocytes (IVM) or in vitro culture (IVC) of embryos impact their cryotolerance. A literature search was performed using three databases, recovering 43 articles for the systematic review, comprising 75 experiments (13 performed in IVM, 62 in IVC) and testing 13 substances. In 39 % of the experiments, an increase in oocyte and/or embryo survival after cryopreservation was reported, in contrast to 48 % exhibiting no effect, 5 % causing negative effects, and 8 % influencing in a dose-dependent manner. Of the 75 experiments extracted during IVM and IVC, 41 quantified the lipid content. Of those that reduced lipid content (n = 26), 50 % increased cryotolerance, 34 % had no effect, 8 % harmed oocyte/embryo survival, and 8 % had different results depending on the concentration used. Moreover, 28 out of the 43 studies were analyzed under a meta-analytical approach at the IVC stage in cattle. There was an improvement in the cryotolerance of bovine embryos when the lipid content was reduced. Forskolin, l-carnitine, and phenazine ethosulfate positively affected cryotolerance, while conjugated linoleic acid had no effect and impaired embryonic development. Moreover, fetal bovine serum has a positive impact on cryotolerance. SOF and CR1aa IVC media improved cryotolerance, while mSOF showed no effect. In conclusion, lipid modulators did not unanimously improve cryotolerance, especially when used in IVM, but presented positive effects on cryotolerance during IVC when reaching lipid reduction.

Combined physical training protects the left ventricle from structural and functional damages in experimental pulmonary arterial hypertension.

BACKGROUND: Under the adverse remodeling of the right ventricle and interventricular septum in pulmonary arterial hypertension (PAH) the left ventricle (LV) dynamics is impaired. Despite the benefits of combined aerobic and resistance physical trainings to individuals with PAH, its impact on the LV is not fully understood. OBJECTIVE: To test whether moderate-intensity combined physical training performed during the development of PAH induced by MCT in rats is beneficial to the LV's structure and function. METHODS: Male Wistar rats were divided into two groups: Sedentary Hypertensive Survival (SHS, n = 7); and Exercise Hypertensive Survival (EHS, n = 7) to test survival. To investigate the effects of combined physical training, another group of rats were divided into three groups: Sedentary Control (SC, n = 7); Sedentary Hypertensive (SH, n = 7); and Exercise Hypertensive (EH, n = 7). PAH was induced through an intraperitoneal injection of MCT (60 mg/kg). Echocardiographic evaluations were conducted on the 22nd day after MCT administration. Animals in the EHS and EH groups participated in a combined physical training program, alternating aerobic (treadmill running: 50 min, 60% maximum running speed) and resistance (ladder climbing: 15 climbs with 1 min interval, 60% maximum carrying load) exercises, one session/day, 5 days/week for approximately 4 weeks. RESULTS: The physical training increased survival and tolerance to aerobic (i.e., maximum running speed) and resistance (i.e., maximum carrying load) exertions and prevented reductions in ejection fraction and fractional shortening. In addition, the physical training mitigated oxidative stress (i.e., CAT, SOD and MDA) and inhibited adverse LV remodeling (i.e., Collagen, extracellular matrix, and cell dimensions). Moreover, the physical training preserved the amplitude and velocity of contraction and hindered the reductions in the amplitude and velocity of the intracellular Ca2+ transient in LV single myocytes. CONCLUSION: Moderate-intensity combined physical training performed during the development of MCT-induced PAH in rats protects their LV from damages to its structure and function and hence increases their tolerance to physical exertion and prolongs their survival.

Heavy metals bioaccumulation in free-ranging South American rattlesnakes (Crotalus durissus) in Southeastern Brazil.

Anthropogenic activities are the main sources of soil, air, and water pollution by metals, including cadmium (Cd), lead (Pb), chromium (Cr), the metalloid arsenic (As), magnesium (Mg), zinc (Zn), and copper (Cu). The goal of this study was to assess the presence and concentration of toxic (As, Cd, Pb, and Cr) and essential metals (Mg, Zn, and Cu) in the liver and kidneys from 96 free-ranging rattlesnakes (Crotalus durissus) from Minas Gerais (Brazil). Bioaccumulation of Cd and Pb were significantly higher in males and heavier rattlesnakes (those with body weight above the average of the study population). Average ± standard deviations of Cd, Pb, Cr, Cu, Mg, Zn, and As in the general population (n = 96) were 3.19 ± 2.52; 5.98 ± 8.49; 0.66 ± 1.97; 3.27 ± 2.85; 776.14 ± 2982.92; 27.44 ± 29.55; and 0.32 ± 1.46; respectively. Bioaccumulation of some metals correlated positively with changes in hematologic and serum biochemical parameters. Results of this study were contrasted with previous studies assessing metal bioaccumulation in other species of terrestrial or aquatic snakes. Considering their position in the food chain and the broad range of bioaccumulation of both toxic and essential metals observed in this study, rattlesnakes may function as highly relevant biological sentinels for environmental pollution.

Impact of Early Arsenic Exposure on the Mineral Content and Oxidative Status of the Liver and Kidney of Pubescent and Adult Rats.

This study evaluated the effect of prepubertal arsenic exposure in the liver and kidney of pubescent rats and their reversibility 30 days after arsenic withdrawal. Male pups of Wistar rats (21 days old) were divided into two groups (n = 20/group): control animals received filtered water, and exposed rats received 10 mg L-1 arsenic from postnatal day (PND) 21 to PND 51. The liver and kidney of 52 days old rats (n = 10/group) were examined to investigate the effects of arsenic on micromineral content, antioxidant enzyme activity, histology, and biochemistry parameters. The other animals were kept alive under free arsenic conditions until 82 days old and further analyzed by the same parameters. Our results revealed that 52-day-old rats increased arsenic content in their liver and arsenic and manganese in their kidney. In those animals, glycogen and zinc content and catalase activity were reduced in the liver, and the selenium content decreased in the kidney. Thirty days later, arsenic reduced the manganese and iron content and SOD and CAT activity in the liver of 82-day-old rats previously exposed to arsenic, while glycogen and selenium content decreased in their kidney. In contrast, PND 82 rats exhibited higher retention of copper in the liver, an increase in iron and copper content, and CAT and GST activity in the kidney. Significant histological alterations of liver and kidney tissues were not observed in rats of both ages. We conclude that arsenic-induced toxicity could alter differently the oxidative status and balance of trace elements in pubertal and adult rats, demonstrating that the metalloid can cause effects in adulthood.

Effect of Induced Pulmonary Arterial Hypertension on Testicular Parameters of Wistar Rats Subjected to Resistance Exercise Training.

Pulmonary arterial hypertension (PAH) is characterized by elevated arterial pressure and vascular resistance. PAH may cause alterations in the microcirculation of several organs, including the kidney, liver, brain, and testes. However, it remains unclear whether monocrotaline-induced PAH exerts detrimental effects on animal testes. Thus, we analyzed the impact of PAH on testicular morphology and function. Additionally, we investigated the effect of resistance exercise training (RT) on testicular parameters in PAH rats. Eight healthy Wistar rats and eight PAH rats were subjected to RT training for 30 days; the other PAH and healthy rats (n = 8/group) did not exercise. PAH rats had lower reproductive organ weight, serum testosterone levels, testicular glucose, and nitric oxide (NO) levels, Leydig cell parameters, tubular morphometry, germ cell counts, and daily sperm production than healthy animals did. The practice of RT attenuated the negative impact of PAH on the relative weights of the testes and epididymides, Leydig cell number, nuclear volume, testicular NO levels, and seminiferous epithelium architecture. Moreover, RT positively influenced testosterone levels in PAH animals. We conclude that PAH exerts deleterious effects on testicular histology and function. However, RT can be beneficial to the PAH-affected testicular parameters.

Investigating the effects of mesotrione/atrazine-based herbicide on honey bee foragers.

A mixture of the herbicides mesotrione and atrazine (Calaris®) is a widely used herbicide in agriculture in several countries. However, the possible toxicological effects of this formulation on non-target organisms require investigation. In this study, the effects of acute oral exposure to Calaris® were evaluated in Apis mellifera foragers. The effect of seven different concentrations of Calaris® on survival and sucrose consumption was studied, while the recommended concentration for field use (FC) and its 10× dilution (0.1 FC) were used to assess overall locomotor activity, respiratory rate, flight, midgut morphology, oxidative and nitrosative stresses, and hemocyte counting. The exposure to FC or 0.1 FC decreased locomotor activity and induced damage to the midgut epithelium. Additionally, the two tested concentrations reduced superoxide dismutase activity, nitric oxide levels, and total hemocyte count. FC also increased malondialdehyde content and 0.1 FC increased respiratory rate and decreased the proportion of prohemocytes. Overall, our findings evidenced significant harmful effects on A. mellifera foragers resulting from the ingestion of the Calaris® herbicide.

The effects of eugenol on histological, enzymatic, and oxidative parameters in the major salivary glands and pancreas of healthy male Wistar rats.

OBJECTIVE: We evaluated the effects of eugenol on histological, enzymatic, and oxidative parameters in the pancreas, parotid, submandibular, and sublingual glands of healthy male rats. DESIGN: Twenty-four adult Wistar rats were assigned into four groups (n = 6/group). Control rats received 2% Tween-20 (eugenol vehicle), whereas the other animals received 10, 20, and 40 mg kg-1 eugenol through gavage daily for 60 d. Major salivary and pancreatic glands were weighed and preserved fixed for microscopic analysis and frozen for in vitro assays. RESULTS: Eugenol did not alter glands' weight and serum amylase activity regardless of the concentration. The highest dose of eugenol caused an increase in pancreatic amylase activity and a reduction of lipase activity from serum and pancreas. Eugenol at 40 mg kg-1 diminished the activity of SOD and FRAP in the submandibular gland and CAT and FRAP in the sublingual gland. However, it did not exert any effect on GST regardless of the gland. Additionally, 40 mg kg-1 eugenol increased MDA levels in pancreatic, parotid, and submandibular glands and NO levels in the sublingual. The concentrations of eugenol induced distinct responses in the glands regarding the activity of Na+/K+, Mg2+, and total ATPase activity. They also affected histomorphometrical and histochemistrical parameters in the submandibular gland only. CONCLUSIONS: Results indicated that 40 mg kg-1 eugenol altered most of the biochemical and oxidatived parameters of digestive glands. Only submandibular glands presented histological changes after eugenol exposure suggesting potential implications for its function.

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.

Post-thaw quality of ram sperm frozen with different concentrations of low-density lipoproteins associated with non-enzymatic antioxidants.

The cryopreservation reduces ram sperm quality, decreasing the pregnancy rate of ewes inseminated with thawed sperm. Hence, we aimed to improve the post-thaw quality of ram sperm replacing egg yolk on Tris-Glucose extender with different concentrations of LDL (2 or 8%), associated with the addition of 10 mM non-enzymatic antioxidants (ascorbic acid, hydroxytoluene butylate, ascorbyl palmitate, and trehalose). Semen samples were collected from six rams, split into different treatments, and frozen. After thawing, kinematic (CASA), structural (propidium iodide and carboxyfluorescein diacetate) and functional (hypoosmotic test) sperm membrane integrity was assessed. Total motility, VCL, and LIN were also assessed in thawed samples during 3 h of incubation (38 °C). The results showed that hydroxytoluene butylate at 10 mM in Tris-Glucose extender with 8% LDL improved velocity parameters immediately post-thaw compared with Tris-Glucose egg yolk extender, as well as prevented the reduction of total motility and VCL after incubation. There was no benefit of adding ascorbic acid and trehalose. Moreover, for the first time, it was shown the motility impairment promoted by ascorbyl palmitate to ram sperm.

Bioinsecticide spinosad poses multiple harmful effects on foragers of Apis mellifera.

There are multifactorial causes for the recent decline in bee populations, which has resulted in compromised pollination and reduced biodiversity. Bees are considered one of the most important non-target insects affected by insecticides used in crop production. In the present study, we investigated the effects of acute oral exposure to spinosad on the survival, food consumption, flight behavior, respiration rate, activity of detoxification enzymes, total antioxidant capacity (TAC), brain morphology, and hemocyte count of Apis mellifera foragers. We tested six different concentrations of spinosad for the first two analyses, followed by LC50 (7.7 mg L-1) for other assays. Spinosad ingestion decreased survival and food consumption. Exposure to spinosad LC50 reduced flight capacity, respiration rate, and superoxide dismutase activity. Furthermore, this concentration increased glutathione S-transferase activity and the TAC of the brain. Notably, exposure to LC50 damaged mushroom bodies, reduced the total hemocyte count and granulocyte number, and increased the number of prohemocytes. These findings imply that the neurotoxin spinosad affects various crucial functions and tissues important for bee performance and that the toxic effects are complex and detrimental to individual homeostasis.

Long-term reproductive effects in male rats prenatally exposed to sodium arsenite.

Arsenic is an environmental toxicant known to be a carcinogen and endocrine disruptor. Maternal exposure to arsenic has been associated with fetus malformation and reproductive disorders in male offspring. However, it is unclear the extent to which those effects remain during postnatal development and adulthood. Therefore, this study aimed to investigate the long-term effects of prenatal arsenic exposure on reproductive parameters of male offspring at peripubertal and adult periods. Pregnant female Wistar rats were exposed to 0 or 10 mg/L sodium arsenite in drinking water from gestational day 1 (GD 1) until GD 21 and male pups were analyzed at postnatal day 44 (PND 44) and PND 70. We observed that some reproductive parameters were affected differently by arsenic exposure at each age evaluated. The body and reproductive organs weights, as well as testicular and epididymal morphology were strongly affected in peripubertal animals and recovered at adult period. On the other hand, the antioxidant genes expression (SOD1, SOD2, CAT and GSTK1) and the endogenous antioxidant system were affected in the testes and epididymides from both peripubertal and adult rats. Finally, an impairment in daily sperm production and in sperm parameters was observed in adult animals. Taken together, our findings show that prenatal arsenic exposure affected reproductive parameters of peripubertal and adult male rats mainly due to oxidative stress. Collectively, those alterations may be affecting fertility potential of adult animals.

Does environmental pollution affect male reproductive system in naturally exposed vertebrates? A systematic review.

Due to environmental contamination, the environment constantly receives pollutants from various anthropic actions. These pollutants put ecological health at risk due to contamination and accumulation in living organisms, including wild animals and humans. Exposure can cause physiological, morphological, and behavioral changes in living beings. In this context, laboratory studies have frequently investigated how environmental contaminants affect the male reproductive system and gametes. However, few studies have examined how these contaminants affect male reproduction in naturally exposed animals. To better understand this topic, we conducted a systematic review of the effects of exposing male vertebrate animals to polluted environments on their reproductive functions. After an extensive search using the PubMed/MEDLINE, Scopus, and Web of Science databases, 39 studies met our inclusion criteria and were eligible for this review. This study showed that reproductive damages were frequent in fishes, amphibians, reptiles, birds, and mammals exposed to contaminated environments. Wild animals are exposed mainly to endocrine-disrupting compounds (EDCs), toxic metals, and radiation. Exposure to pollutants causes a reduction in androgen levels, impaired spermatogenesis, morphological damage to reproductive organs, and decreased sperm quality, leading to reduced fertility and population decline. Although several species have been studied, the number of studies is limited for some groups of vertebrates. Wildlife has proven valuable to our understanding of the potential effects of environmental contaminants on human and ecosystem health. Thus, some recommendations for future investigations are provided. This review also creates a baseline for the understanding state of the art in reproductive toxicology studies.

Contributions of seminal plasma proteins to fertilizing ability of bull sperm: A meta-analytical review.

Seminal plasma is a dynamic, intricate combination of fluids from the testicles, epididymides, seminal vesicles, bulbourethral glands, and prostate, containing molecules that modulate sperm functions, post-fertilization events, and the female reproductive tract physiology. Significant variations in sperm parameters and fertility status of bulls relate to differences in the seminal plasma proteome. In this framework, a meta-analytical study was conducted examining 29 studies (published between 1990 and 2021) to ascertain the effects of seminal fluid proteins on parameters associated with bull fertility and the influence of distinct methodologies on such effects. Our results revealed that seminal proteins ameliorate sperm parameters, such as motility, integrity, capacitation, and fertilizing ability, and favours sperm protection. Seminal binder of sperm proteins and beta-defensin 126 highly favoured sperm protection when cells were collected from the epididymis by retrograde flux and analysed under room temperature conditions. Furthermore, seminal proteins improved the motility and quality of Bos taurus sperm collected by artificial vagina, mainly in the presence of heparin-binding proteins. The key limitations faced by this meta-analysis were the paucity of studies evaluating the effects of whole seminal fluid proteins and the limited number of studies conducted in vivo. In conclusion, the present meta-analytical study confirms that seminal proteins improve fertility-related parameters in the bovine species. However, methodological strategies used by authors are diverse, with distinct endpoints and methods. Thus, the translational aspects of seminal plasma research should be taken into consideration to precisely define how seminal proteins can be harnessed to advance reproductive biotechnology.

Comparison Between Gynecological Examination Methods and Sample Collection Techniques for the Diagnosis of Endometritis in Subfertile Mares.

Endometritis is a relevant cause of subfertility in mares. However, the accurate diagnosis, essential for effective treatment, can be difficult due to the variability of results and interpretations resulting from different examination methods and sample collection techniques. The present work compared gynecological evaluation methods and sample collection techniques to diagnose endometritis in subfertile mares. Forty animals with a history of subfertility were selected for gynecological evaluation using clinical methodologies, such as perineal conformation, transrectal palpation and ultrasonography, vaginoscopy, and digital examination of the cervix. In addition, we performed laboratory analyses, including uterine microbiological culture and endometrial cytology and histology, of which the latter is the gold standard for the diagnosis of endometritis. Samples were collected for microbiological culture and endometrial cytological evaluations using three different techniques: a commercial cytobrush/swab collector, low-volume uterine flush, and a new tested technique, by flush the fragment resulting from the endometrial biopsy. Transrectal palpation and ultrasound showed the best results among clinical examinations. However, they were less efficient in laboratory tests of endometrial cytology and uterine microbiological culture, in which the latter showed the highest sensitivity and specificity for endometritis compared with endometrial histology. The use of multiple results from different methods has also proved to be an effective alternative for diagnosis. Among the techniques used to collect endometrial material for cytology and microbiological culture, the most effective and practical in this study was the commercial cytobrush/swab collector.

Immunomodulatory activity of trifluoromethyl arylamides derived from the SRPK inhibitor SRPIN340 and their potential use as vaccine adjuvant.

The serine/arginine-rich protein kinases (SRPK) specifically phosphorylate their substrates at RS-rich dipeptides, which are abundantly found in SR splicing factors. SRPK are classically known for their ability to affect the splicing and expression of gene isoforms commonly implicated in cancer and diseases associated with infectious processes. Non-splicing functions have also been attributed to SRPK, which highlight their functional plasticity and relevance as therapeutic targets for pharmacological intervention. In this sense, different SRPK inhibitors have been developed, such as the well-known SRPIN340 and its derivatives, with anticancer and antiviral activities. Here we evaluated the potential immunomodulatory activity of SRPIN340 and three trifluoromethyl arylamide derivatives. In in vitro analysis with RAW 264.7 macrophages and primary splenocytes, all the compounds modulated the expression of immune response mediators and antigen-presentation molecules related to a tendency for M2 macrophage polarization. Immunization experiments were carried out in mice to evaluate their potential as vaccine immunostimulants. When administrated alone, the compounds altered the expression of immune factors at the injection site and did not produce macroscopic or microscopic local reactions. In addition, when prepared as an adjuvant with inactivated EHV-1 antigens, all the compounds increased the anti-EHV-1 neutralizing antibody titers, a change that is consistent with an increased Th2 response. These findings demonstrate that SRPIN340 and its derivatives exhibit a noticeable capacity to modulate innate and adaptative immune cells, disclosing their potential to be used as vaccine adjuvants or in immunotherapies.

Could metal exposure affect sperm parameters of domestic ruminants? A meta-analysis.

During the last decade, environmental toxicants have been considered a potential cause for declining sperm quality. Toxic metals are not easily degraded and may accumulate along the food chain, which may negatively impact the semen quality of animals. In this framework, we conducted a meta-analysis to determine whether exposure to Al, As, Cd, Cr, Co, Cu, Fe, Mg, Mn, Hg, Ni, and Pb affects sperm and andrological parameters of domestic ruminants. We extracted 217 independent comparisons from 39 published articles selected from PubMed, Web of Science, and Scopus. Our findings showed that metal exposure reduced sperm viability (d++ = - 1.04, df 51, CI - 1.47 to - 0.61) and motility (d++ = - 0.83, df 104, CI -1.19 to - 0.51) by increasing oxidative metabolites production (d++ = 2.98, df 20, CI 1.95-0.11). Sperm viability and motility were affected by Cd, As, Hg, and Fe contamination. Metal poisoning impaired andrological parameters (d++ = - 0.83, df 17, CI - 1.10 to - 0.02) after arsenic intake using 3 and 5 mg L-1 orally. Detrimental effects on spermatozoa were mostly observed after in vitro incubation with metals using concentrations < 2.99 mg L-1 up to 24 h. The review limitations were the heterogeneity of methodologies used in the studies and absence of investigations focused on the effect of Al, Co, Cr, Mg, and Ni exposure on sperm parameters in ruminants. Nevertheless, our findings contribute to understanding the impact of metal exposure on reproductive parameters in ruminants, with potential damage to their fertility.

Eugenol reduces serum testosterone levels and sperm viability in adult Wistar rats.

Eugenol is the main constituent of clove extract. It is a remarkably versatile molecule incorporated as a functional ingredient in several food products and widely applied in the pharmaceutical industry. Men consume natural products enriched with eugenol for treating sexual disorders and using as aphrodisiacs. Nevertheless, there is no information about the impact of eugenol intake on the reproductive parameters of healthy males. Therefore, we provided 10, 20, and 40 mg kg-1 pure eugenol to adult Wistar rats for 60 days. Testis, epididymis, and spermatozoa were analyzed under microscopic, biochemical, and functional approaches. This phenolic compound did not alter testicular and epididymal biometry and microscopy. However, 20 and 40 mg kg-1 eugenol reduced serum testosterone levels. The highest dose altered lactate and glucose concentrations in the epididymis. All the eugenol concentrations diminished CAT activity and MDA levels in the testis and increased FRAP and CAT activity in the epididymis. Epididymal sperm from rats receiving 10, 20, and 40 mg kg-1 eugenol presented high Ca2+ ATPase activity and low motility. In conclusion, eugenol at low and high doses negatively impacted the competence of epididymal sperm and modified oxidative parameters in male organs, with no influence on their microscopy.

The SRPK inhibitor N-(2-(piperidin-1-yl)-5-(trifluoromethyl)phenyl) isonicotinamide (SRPIN340) increases the immune response against metastatic melanoma in mice.

Cancers have a strong relationship with immune cells in their microenvironment, which significantly influences tumor proliferation and progression. Thus, pharmacological strategies that stimulate the immune system to combat tumor cells are promising for better therapeutic efficacy. Deregulated expression of the splicing regulatory serine arginine protein kinases (mostly SRPK1 and SRPK2) has been found in different cancer types, leading to the expression of isoforms related to tumor growth and metastasis. The microenvironment of melanoma exhibits a strong presence of immune cells, which significantly influences tumor progression, and around 50% of cutaneous melanoma patients benefit from targeted immunotherapy. Here, we analyzed human malignant melanoma single-cell gene expression data and observed that SRPK1/2 overexpression correlates with immune system pathway alterations. In further analysis, we observed an increased presence of immune cells in biopsies from mice bearing metastatic melanoma treated with SRPIN340, a well-known SRPK1/2 pharmacological inhibitor. Local treatments increased the expression of proinflammatory cytokines at the tumor lesions and the activity of the spleen, accompanied by reduced pulmonary metastasis foci, edema formation, and alveolar congestion. In in vitro assays, SRPIN340 also potentiated immunological susceptibility, by increasing the expression of the antigen presenting MHCI and MHCII molecules and by increasing the ability of B16F10 cells to attract splenic cells in transwell assays. Taken together, these results reveal that the antimetastatic effect of SRPIN340 can also involve an increased immune response, which suggests additional functional clues for SRPKs in tumor biology.

High doses of eugenol cause structural and functional damage to the rat liver.

Eugenol is a phenolic compound found in clove extract and extensively used in traditional medicine. It is unclear whether its intake can cause positive or negative effects on liver morphology and physiology in healthy individuals. Thus, we aimed to evaluate liver parameters of rats treated with 10, 20, and 40 mg kg-1 eugenol. After 60 days of treatment, liver samples were collected and analyzed by biometric, histological, biochemical, and oxidative analyses. Our results showed that 10, 20, and 40 mg kg-1 eugenol did not alter body and liver weights, serum and hepatic ALT levels and catalase, glutathione-s-transferase, total, Ca2+, and Mg2+ ATPases activities in treated animals. However, 20 and 40 mg kg-1 eugenol reduced Na+/K+ ATPase pump activity and blood glucose levels. They also increased hepatic glycogen content, superoxide dismutase activity, ferric reducing antioxidant power, and nitric oxide and malondialdehyde levels. Still, 20 and 40 mg kg-1 eugenol caused structural and functional damage to the liver tissue of eugenol-treated rats. We concluded that 10 mg kg-1 eugenol is a safe dose for consumption in long-term treatment for rats. Doses higher than 20 mg kg-1 lead to hepatic damage that can impair vital processes of liver functionality.

Testicular morphometry as a tool to evaluate the efficiency of immunocastration in lambs.

This study aimed to evaluate the efficiency of immunocastration in lambs using testicular morphometry. Thirty lambs were randomly divided into two treatments (subcutaneous administration of 1.0 mL and 0.5 mL of an anti-GnRH vaccine) and a control group (1.0 mL saline solution). The animals were vaccinated at four months of age, received a second dose 30 days later, and were slaughtered 90 days after the first vaccine dose. After slaughter, testicles were collected, and samples were removed for histological processing and evaluation of testicular morphometric parameters. Analysis of variance, Tukey's test, and Kruskal-Wallis test were performed, with a 5% level of significance. There was a reduction in testicular weight, gonadosomatic index, seminiferous tubule diameter, germinal epithelium height, leydigosomatic index, and total tubule length. The total length per testicular gram increased in the immunocastrated group. Intrinsic spermatogenesis yield, Sertoli cell indices, and estimates of sperm and Sertoli cell production were reduced in the immunized groups (P < 0.05). The anti-GnRH vaccine in lambs at doses of 1.0 mL and 0.5 mL is sufficient to promote immunocastration, verified through severe changes in testicular morphometry from animals.