Male Animals Research Articles

Can exercise prevent the age‐related decline in adaptive homeostasis? Evidence across organisms and tissues

Can exercise prevent the age‐related decline in adaptive homeostasis? Evidence across organisms and tissues

Differences and the impact of G protein-coupled estrogen receptor deletion in the vascular and renal response to aldosterone

We recently showed that the G protein-coupled estrogen receptor (GPER) protects female mice from arterial stiffening, an independent predictor of cardiovascular mortality. Previous work indicates that GPER may directly bind aldosterone and/or dimerize with the mineralocorticoid receptor. Therefore, we hypothesized that GPER may promote aldosterone-induced renal and vascular effects, particularly in male animals where less estrogen is available to activate GPER. Male (M) and female (F) wildtype (wt) and global GPER knockout (ko) mice were used between 6-7 months of age. Aldosterone was delivered to all mice subcutaneously via osmotic mini pumps at a dose of 600 μg/kg/day and drinking water was replaced with saline for 28 days. At the end of the study, tail-cuff systolic blood pressure (SBP) and metabolic cage data were collected. Data were analyzed by 2-way ANOVA followed by Sidak’s test and correlation analysis. In basal conditions, GPER ko increased systolic SBP in males but not females. After aldosterone-salt treatment, SBP was similar in Mwt versus Mko but was significantly higher in Fko versus Fwt (Pinteraction=0.002; 148 ± 12 vs. 108 ± 2 mmHg; P<0.001). Similarly, proteinuria was similar in males but greater in Fko than Fwt (Pinteraction=0.01; 6.0 ± 1.5 vs. 2.4 ± 0.4 mg/day; P=0.04), indicating greater kidney injury. In summary, GPER deletion in females revealed enhanced blood pressure and renal damage in response to an aldosterone-salt challenge. These findings do not support a role for GPER in promoting the impact of aldosterone on vascular and renal damage in either sex. In contrast, GPER protected female mice from the negative impact of aldosterone-salt on blood pressure and kidney function. More studies are needed to understand the interactions of GPER and aldosterone in the kidney of both males and females. This work was supported by the National Institutes of Health (HL133619 and AG071746 to SL, HL155841 to BO) and the American Heart Association (BV and IK) This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Sex differences in cachexia outcomes, anabolic signalling and BCAA metabolism following chemotherapy

Sex is a risk factor for cancer. Affecting nearly 80% of cancer patients is cachexia, a body and skeletal muscle-wasting syndrome. Poor nutritional status, tumour related factors and chemotherapy contribute to cachexia. Although negative effects of chemotherapy on skeletal muscle are documented, the majority of studies are completed in male animals. Although the branched-chain amino acids (BCAA: leucine, isoleucine and valine) activate anabolic signalling in skeletal muscle and have been shown to reduce some of the effects of cachexia, BCAA nutritional support does not reverse cachexia and few studies have investigated the effects of chemotherapy on BCAA metabolism in skeletal muscle. Therefore, the objective of this study is to compare the effects of chemotherapy on cachexia outcomes and BCAA metabolism between male and female mice. Three-month-old CD2F1 male and female mice were treated with either the chemotherapy drug cocktail folfiri (50mg/kg 5-fluorouracil (5FU), 90mg/kg Leucovorin and 24mg/kg CPT11) (drug) or vehicle (10% DMSO in saline) for 6-weeks. Within sex, drug treatment led to reductions in body and skeletal muscle weights. Between sex, drug-treated female mice lost more body and gastrocnemius muscle weight compared to drug-treated males. Within sex, drug treated animals exhibited reductions in anabolic signalling, specifically AKT, S6K1, S6 and 4E-BP1. Between sex, drug-treated females mice showed greater loss of anabolic signalling compared to drug-treated males. Within sex, the drug cocktail reduced amino acid transporters SNAT1 and LAT1, while reducing BCAA concentrations in the skeletal muscle. Drug-treated female mice exhibited greater loss of LAT1 compared to male-drug treated mice. However, male-drug treated mice had greater decreases in BCAA concentrations compared to female drug-treated mice. In addition, minimal differences were found within sex for key enzymes involved in BCAA metabolism. However, the inhibitory phosphorylation of branched-chain alpha-ketoacid dehydrogenase complex (BCKD) was elevated only in drug-treated male animals. Lastly, within sex, drug treatment showed elevated levels of BCAA concentrations in the liver, but only male drug-treated mice showed elevated plasma BCAAs compared to male vehicle-treated mice. Our data suggests a link between muscle atrophy and decreased BCAA metabolism during chemotherapy. We found that drug-treated female mice had worsened outcomes for cachexia measures such as body, skeletal muscle weight and anabolic signalling compared to drug-treated male mice. However, drug-treated males exhibited greater decreases in BCAA concentrations and their metabolism compared to drug-treated females. Findings from this study suggest that altered availability and metabolism of the BCAAs may contribute to muscle wasting differently in males and females during chemotherapy and interventions that correct such alterations may need to consider sex in order to better manage cachexia. Nserc, York University This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

The action of phosphodiesterase-5 inhibitors on β-amyloid pathology and cognition in experimental Alzheimer's disease: A systematic review.

Alzheimer's disease (AD) is the most frequent cause of dementia worldwide. The etiology of AD is partially explained by the deposition of β-amyloid in the brain. Despite extensive research on the pathogenesis of AD, the current treatments are ineffective. Here, we systematically reviewed studies that investigated whether phosphodiesterase 5 inhibitors (PDE5i) are efficient in reducing the β-amyloid load in hippocampi and improving cognitive decline in rodent models with β-amyloid accumulation. We identified ten original studies, which used rodent models with β-amyloid accumulation, were treated with PDE5i, and β-amyloid was measured in the hippocampi. PDE5i was efficient in reducing the β-amyloid levels, except for one study that exclusively used female rodents and the treatment did not affect β-amyloid levels. Interestingly, PDE5i prevented cognitive decline in all studies. This study supports the potential therapeutic use of PDE5i for the reduction of the β-amyloid load in hippocampi and cognitive decline. However, we highlight the importance of conducting additional experimental studies to evaluate the PDE5i-related molecular mechanisms involved in β-amyloid removal in male and female animals.

Biological effects of exposure to 2650 MHz electromagnetic radiation on the behavior, learning, and memory of mice.

With the development of communication technology, the public is paying increasing attention to whether electromagnetic radiation is harmful to health. Mobile phone communication has entered the 5G era, and there are almost no reports on electromagnetic radiation at 2650 MHz. Therefore, it is necessary to evaluate the risk of adverse effects of 5G mobile phone EMR exposure on the human brain. Male animals were continuously exposed to 2650 MHz-EMR for 28 days with a whole-body averaged specific absorption rate (WBSAR) of 2.06 W/kg for 4 h per day. Mouse behavior was assessed using the open-field test (OFT), elevated-plus maze (EPM), and tail suspension test (TST). The Morris water maze (MWM), HE staining, and TUNEL staining were used to evaluate the spatial memory ability and pathological morphology of hippocampal dentate gyrus cells. Additionally, the expression levels of brain-derived neurotrophic factor (BDNF), aminobutyric acid (GABA), and glucocorticoid (GR) in the hippocampus were detected by western blotting and immunohistochemistry, while the corticosterone (CORT) level in serum was detected by ELISA. In the OFT, the total distance traveled, central distance traveled, and residence time significantly decreased in the EMR exposure group (p < .05). In EPM, the percentage of the number of times to open the arm and the percentage of time to open the arm significantly decreased in the EMR exposure group. However, in the TST, the two groups had no significant difference in the 4-min immobility time. In the MWM, the escape latency of the EMR exposure group was shorter than that of the control group, with no significant difference. Furthermore, CORT levels in serum were significantly increased in the EMR exposure group (p < .05), while the expression of BDNF and GR proteins in the hippocampus was reduced (p < .05), but there was no significant difference in GABA expression. Our results indicate that exposure to 2650 MHz-EMR (WBSAR: 2.06 W/kg, 28 days, 4 h per day) had no significant effect on the spatial memory ability of mice (in comparison to little effect). The exposure may be associated with anxiety-like behavior in mice but not related to depression-like behavior in mice.

Soda intake influences phenotype, antioxidants and inflammatory status in high protein-fed wistar rats

An increasing population of people, especially young adults who exercise, consume high protein diets along with carbonated drinks. While there are numerous studies on the effect of high protein diets, there is a need to understand how protein diets in combination with carbonated drinks impact physiology. In order to assess these effects on wistar rats’ phenotype, antioxidants and inflammatory profiles, 64 wistar rats were divided into dietary groups of 8 male and 8 female animals each. The animals were fed standard diet as control (chow), chow and carbonated soda, a high protein diet (48.1% energy from protein) and a high protein diet with carbonated soda according to their groups.Body measurements, blood glucose levels, serum insulin levels, lipid peroxidation, antioxidant activity, adipokines and inflammatory markers concentrations were all determined. At the end of the study, body measurements, inflammatory markers and adipokine concentration were increased in animals fed the high protein diet and high protein-soda diet. There was a decrease in antioxidant and lipid peroxidation levels in protein fed male and female animals but those fed protein in combination with soda had increased lipid peroxidation levels.In conclusion, high protein diet in combination with carbonated soda impacts physiology differently from a high protein diet alone, and may stimulate weight gain, oxidative stress and HPD-related inflammation in Wistar rats.

Palmitic acid-modified GnRH-Th epitope peptide immunocastration vaccine (W/O/W adjuvant) can effectively ensure the castration and reduce the smelly smell in boars.

Recent studies have demonstrated the effectiveness of Gonadotropin-releasing hormone (GnRH) in inhibiting testicular growth and development in male animals to achieve castration while improving the meat quality of various livestock species, including cattle, sheep, goats, and pigs. In this research, a GnRH-Th vaccine was synthesized using the Fmoc solid-phase synthesis technique, and the T helper (Th) antigen was modified with palmitic acid to improve its efficacy. The vaccine was then coated with a water-in-oil-in-water adjuvant to improve stability and safety. After passing safety and stability tests, the vaccine was administered to 13-week-old boars. The results showed that it was stable, safe, and effective for up to 15 months. Moreover, the vaccine did not negatively affect the growth rate and body weight of the pigs. The palmitic acid-modified "GnRH-Th epitope peptide immunocastration vaccine (Water-in-Oil-in-Water (W/O/W)) effectively reduced the testosterone concentration and achieved castration. The concentration of androstenone and skatole hormones significantly decreased, leading to improved meat quality in the boars. The boars were then slaughtered at 33 weeks of age, and the results showed that the meat quality of the vaccinated boars was superior to that of the non-vaccinated control group (p < 0.05). This study demonstrated that GnRH can safely and effectively achieve immune castration in boars after coupling T cell epitopes, palmitic acid modification and W-O-W coating. Provide a better method for the further development of GnRH and the realization of animal welfare.

Sex and estrous cycle affect experience-dependent plasticity in mouse primary visual cortex.

Sex hormones can affect cellular physiology and modulate synaptic plasticity, but it is not always clear whether or how sex-dependent differences identified in vitro express themselves as functional dimorphisms in the brain. Historically, most experimental neuroscience has been conducted using only male animals and the literature is largely mute about whether including female mice in will introduce variability due to inherent sex differences or endogenous estrous cycles. Though this is beginning to change following an NIH directive that sex should be included as a factor in vertebrate research, the lack of information raises practical issues around how to design experimental controls and apply existing knowledge to more heterogeneous populations. Various lines of research suggest that visual processing can be affected by sex and estrous cycle stage. For these reasons, we performed a series of in vivo electrophysiological experiments to characterize baseline visual function and experience-dependent plasticity in the primary visual cortex (V1) of male and female mice. We find that sex and estrous stage have no statistically significant effect on baseline acuity measurements, but that both sex and estrous stage have can modulate two mechanistically distinct forms of experience dependent cortical plasticity. We also demonstrate that resulting variability can be largely controlled with appropriate normalizations. These findings suggest that V1 plasticity can be used for mechanistic studies focusing on how sex hormones effect experience dependent plasticity in the mammalian cortex.

The circadian clock remains intact, but with dampened hormonal output in heart failure

Circadian (24-h) rhythms are important regulators in physiology and disease, but systemic disease may disrupt circadian rhythmicity. Heart failure (HF) is a systemic disease affecting hormonal regulation. We investigate whether HF affects the rhythmic expression of melatonin and cortisol, main endocrine products of the central clock, and cardiac-specific troponin in patients. We corroborate the functionality of the peripheral clock directly in the organs of translational models, inaccessible in human participants. We included 46 HF patients (71.7% male, median age of 60 years, NYHA class II (32.6%) or III (67.4%), ischemic cardiomyopathy (43.5%), comorbidities: diabetes 21.7%, atrial fibrillation 30.4%), and 24 matched controls. Blood was collected at seven time-points during a 24-h period (totalling 320 HF and 167 control samples) for melatonin, cortisol, and cardiac troponin T (cTnT) measurements after which circadian rhythms were assessed through cosinor analyses, both on the individual and the group level. Next, we analysed peripheral circadian clock functionality using cosinor analysis in male animal HF models: nocturnal mice and diurnal zebrafish, based on expression of core clock genes in heart, kidneys, and liver, every 4h during a 24-h period in a light/darkness synchronised environment. Melatonin and cortisol concentrations followed a physiological 24-h pattern in both patients and controls. For melatonin, acrophase occurred during the night for both groups, with significantly decreased amplitude (median 5.2 vs 8.8, P=0.0001) and circadian variation ([maximum]/[minimum]) in heart failure patients. For cortisol, mesor showed a significant increase for HF patients (mean 331.9 vs 275.1, P=0.017) with a difference of 56.8 (95% CI 10.3-103.3) again resulting in a relatively lower variation: median 3.9 vs 6.3 (P=0.0058). A nocturnal blood pressure dip was absent in 77.8% of HF patients. Clock gene expression profiles (Bmal, Clock, Per, Cry) were similar and with expected phase relations in animal HF models and controls, demonstrating preserved peripheral clock functionality in HF. Furthermore, oscillations in diurnal zebrafish were expectedly in opposite phases to those of nocturnal mice. Concordantly, cTnT concentrations in HF patients revealed significant circadian oscillations. Central clock output is dampened in HF patients while the molecular peripheral clock, as confirmed in animal models, remains intact. This emphasises the importance of taking timing into account in research and therapy for HF, setting the stage for another dimension of diagnostic, prognostic and therapeutic approaches. Hartstichting.

Alterations in cytoskeletal and Ca2+ cycling regulators in atria lacking the obscurin Ig58/59 module.

Obscurin (720-870 kDa) is a giant cytoskeletal and signaling protein that possesses both structural and regulatory functions in striated muscles. Immunoglobulin domains 58/59 (Ig58/59) of obscurin bind to a diverse set of proteins that are essential for the proper structure and function of the heart, including giant titin, novex-3, and phospholamban (PLN). Importantly, the pathophysiological significance of the Ig58/59 module has been further underscored by the discovery of several mutations within Ig58/59 that are linked to various forms of myopathy in humans. We previously generated a constitutive deletion mouse model, Obscn-ΔIg58/59, that expresses obscurin lacking Ig58/59, and characterized the effects of this deletion on cardiac morphology and function through aging. Our findings demonstrated that Obscn-ΔIg58/59 male animals develop severe arrhythmia, primarily manifesting as episodes of junctional escape and spontaneous loss of regular p-waves, reminiscent of human atrial fibrillation, accompanied by significant atrial enlargement that progresses in severity with aging. To comprehensively characterize the molecular alterations responsible for these pathologies, we performed proteomic and phospho-proteomic analyses in aging Obscn-ΔIg58/59 atria. Our studies revealed extensive and novel alterations in the expression and phosphorylation profile of major cytoskeletal proteins, Ca2+ regulators, and Z-disk associated protein complexes in the Obscn-ΔIg58/59 atria through aging. These studies implicate obscurin, particularly the Ig58/59 module, as an essential regulator of the Z-disk associated cytoskeleton and Ca2+ cycling in the atria and provide new molecular insights into the development of atrial fibrillation and remodeling.

Single-cell Genomics: A Quantum Leap toward Understanding Complexity?

Single-cell Genomics: A Quantum Leap toward Understanding Complexity?

Epidemiological analysis of Paramphistomum species in Ruminants in selected Districts of Khyber Pakhtunkhwa (KPK), Pakistan

This epidemiological study was conducted to compare the climatic conditions of four different districts of Khyber Pakhtunkhwa, Pakistan, in terms of the prevalence of parasitic infection Paramphistomum in cows and buffaloes. A total of 2400/cows and buffaloes (200/per month and 50/per district) fecal samples were collected. The samples were examined for Paramphistomum egg contamination by direct microscopic examination and sedimentation methods. The infectivity ratio was 17.5% in cows and 17% in buffaloes; the higher rate of infection was recorded in autumn with 41% in cows and 36% in buffaloes. Whereas the lowest ratio was 6.5% in cows and 7% in buffaloes recorded during spring. It was also noted that a high rate of prevalence was recorded in older animals (higher than 2 years of age) than younger (less than 2 years of age), while gender-wise, it was observed that male animals were more affected than females. Prevalence was higher in cows in Mardan district (19.30%) followed by Nowshera (19.10%) and Swabi (17.80%) districts while lowest in Charsadda district (13.80%) where in buffaloes the higher prevalence was recorded in Swabi (18.80%) followed by Mardan (18.60%) then Charsadda (16.80%) while lowest in Nowshera (13.80%).

Increased male investment in sperm competition results in reduced maintenance of gametes.

Male animals often show higher mutation rates than their female conspecifics. A hypothesis for this male bias is that competition over fertilization of female gametes leads to increased male investment into reproduction at the expense of maintenance and repair, resulting in a trade-off between male success in sperm competition and offspring quality. Here, we provide evidence for this hypothesis by harnessing the power of experimental evolution to study effects of sexual selection on the male germline in the seed beetle Callosobruchus maculatus. We first show that 50 generations of evolution under strong sexual selection, coupled with experimental removal of natural selection, resulted in males that are more successful in sperm competition. We then show that these males produce progeny of lower quality if engaging in sociosexual interactions prior to being challenged to surveil and repair experimentally induced damage in their germline and that the presence of male competitors alone can be enough to elicit this response. We identify 18 candidate genes that showed differential expression in response to the induced germline damage, with several of these previously implicated in processes associated with DNA repair and cellular maintenance. These genes also showed significant expression changes across sociosexual treatments of fathers and predicted the reduction in quality of their offspring, with expression of one gene also being strongly correlated to male sperm competition success. Sex differences in expression of the same 18 genes indicate a substantially higher female investment in germline maintenance. While more work is needed to detail the exact molecular underpinnings of our results, our findings provide rare experimental evidence for a trade-off between male success in sperm competition and germline maintenance. This suggests that sex differences in the relative strengths of sexual and natural selection are causally linked to male mutation bias. The tenet advocated here, that the allocation decisions of an individual can affect plasticity of its germline and the resulting genetic quality of subsequent generations, has several interesting implications for mate choice processes.

The Effect of Tarantula cubensis (Theranekron) on Some Reproductive Parameters in Male Rats

Background: Tarantula cubensis (theranekron), obtained from the poison of Cuban Tarantula, is a homeopathic drug frequently used in animal health. Objectives: The effect of theranekron on reproductive parameters in male animals is unknown. In this study, the effect of theranekron on male reproductive parameters was investigated. Methods: Three different groups of rats were created: control (n=8) (no application), theranekron I (n=8) (subcuteneous injection of 0.3 mg/kg on days 0, 5, and 10), and theranekron II (n=8) (subcuteneous injection of 0.3 mg/kg on days 0, 5, and 10). Control and theranekron I groups were sacrificed at the end of the 4 weeks, and the theranekron II group was sacrificed at the end of the 6 weeks. Sperm motility, density, abnormal rate, DNA damage, and testicular histopathology were examined. Results: No statistical difference was observed in the parameters examined between the study groups (P&gt;0.05). It was determined that the theranekron did not change sperm motility, density and spermatozoa ratio, also no histopathological changes, and sperm DNA damage. Conclusion: There is no harm in using theranekron in male animals during the breeding and nonbreeding seasons.

Prospective analysis of risk factors for perioperative infections in clean and clean-contaminated surgeries in dogs and cats with special reference to perioperative and postoperative antibiotic use

With every surgical procedure there is a risk of postoperative infection (surgical site infection = SSI). This risk of infection can be influenced by various factors, including perioperative antibiotic prophylaxis. In terms of antibiotic stewardship, antibiotics should only be used if there is a proven benefit for the patient. However, this advantage has not yet been conclusively proven, especially for clean and clean-contaminated surgeries. The aim of our study was to document various relevant influencing factors on the infection rate after clean and clean-contaminated surgeries in dogs and cats. In particular, it was documented to what extent a reduced use of antibiotics affects the infection rate in the context of all influencing factors. Over a period of eleven months, 807 clean and clean-contaminated surgeries in dogs and cats were prospectively analyzed with possible influencing factors (gender, ASA classification, underlying endocrinological diseases, duration of anesthesia, duration of surgery, type of surgery, perioperative antibiotic prophylaxis (POA), duration of hospitalization) affecting the infection rate. After surgery all cases were followed up either 30 or 90 days, if implants were used. The effect of the various factors was evaluated using multivariable logistic regression analysis. SSI was detected in 25/664 clean and 10/143 clean-contaminated surgeries. Longer hospitalization, without antimicrobial prophylaxis, and male animals had a significantly higher risk of SSI. In clean surgeries, SSI occurred in 2,3 % of all cases with POA and 5,3 % without POA. The SSI in clean-contaminated was 3,6 % with POA and 9 % without. This difference resulted mainly from the results of osteosynthesis, gastrointestinal and skin surgeries. However, other types of surgeries, such as castrations, neurological interventions, abdominal and thoracic surgeries, and surgeries in the head and neck region, showed comparable infection rates with and without POA.

Unclear fertility-related losses in cattle - a pilot study to evaluate the occurrence of bovine endometrosis

With every surgical procedure there is a risk of postoperative infection (surgical site infection = SSI). This risk of infection can be influenced by various factors, including perioperative antibiotic prophylaxis. In terms of antibiotic stewardship, antibiotics should only be used if there is a proven benefit for the patient. However, this advantage has not yet been conclusively proven, especially for clean and clean-contaminated surgeries. The aim of our study was to document various relevant influencing factors on the infection rate after clean and clean-contaminated surgeries in dogs and cats. In particular, it was documented to what extent a reduced use of antibiotics affects the infection rate in the context of all influencing factors. Over a period of eleven months, 807 clean and clean-contaminated surgeries in dogs and cats were prospectively analyzed with possible influencing factors (gender, ASA classification, underlying endocrinological diseases, duration of anesthesia, duration of surgery, type of surgery, perioperative antibiotic prophylaxis (POA), duration of hospitalization) affecting the infection rate. After surgery all cases were followed up either 30 or 90 days, if implants were used. The effect of the various factors was evaluated using multivariable logistic regression analysis. SSI was detected in 25/664 clean and 10/143 clean-contaminated surgeries. Longer hospitalization, without antimicrobial prophylaxis, and male animals had a significantly higher risk of SSI. In clean surgeries, SSI occurred in 2,3 % of all cases with POA and 5,3 % without POA. The SSI in clean-contaminated was 3,6 % with POA and 9 % without. This difference resulted mainly from the results of osteosynthesis, gastrointestinal and skin surgeries. However, other types of surgeries, such as castrations, neurological interventions, abdominal and thoracic surgeries, and surgeries in the head and neck region, showed comparable infection rates with and without POA.

80 The Need for a Clinical and Translational Science Framework to Bridge Environmental Contamination Data and Male Reproductive Health

OBJECTIVES/GOALS: Although there is ample evidence that environmental contaminants impact reproductive health, the exact mechanisms of action, for the most part, remains unclear. We sought to determine whether known contaminants in Puerto Rico can contribute to the selection of a bioassay to add granularity to geospatial contamination data at the cellular level. METHODS/STUDY POPULATION: A PubMed literature search was conducted: Puerto Rico AND Vieques AND Environmental Contaminants AND Heavy Metals OR Phthalates OR Metals OR PCB OR Air Pollution OR CVOC . Additional inclusion criteria were free full text, English language and year of publication between 2000 to 2022 (n = 244 studies). References that were not related to Puerto Rico and environmental contaminants in air, soil, water, or vegetation were excluded. A second PubMed literature search was conducted to determine whether a clinical link has been established between contaminant exposure and the male reproductive system. Search terms were: heavy metals AND hypospadias OR cryptorchidism NOT female NOT animal NOT review , heavy metals AND male infertility NOT female NOT animal NOT review . The same strategy was used for phthalates. RESULTS/ANTICIPATED RESULTS: We found that 12 out of 15 studies that were conducted in the Archipelago of Puerto Rico between 2000-2022 reported heavy metals- and/or phthalates- contamination in soil and water. We also found that there is a paucity of clinical studies that consider plausible relationships between a given contaminant and congenital conditions or male reproductive function. Specifically, we found that heavy metal exposure has been linked to hypospadias (n=1 study), comorbidity of hypospadias plus cryptorchidism (n= 1 study) or male infertility (n=14 studies). Phthalates exposure has been linked to comorbidity of hypospadias and cryptorchidism (n=1 study) or male infertility (n=1 study). Male subfertility has been overlooked so far. We noted that Sertoli cell dysfunction has been linked to all of these conditions. DISCUSSION/SIGNIFICANCE: The geography of Puerto Rico provides an opportunity to close the gap in knowledge between environmental contamination and male reproductive health. Based on our findings, we propose that the use of a bioassay with an immortalized Sertoli cell line can uncover the cellular processes that may be affected in male reproduction upon contaminant exposure.

RAGE is a critical factor of sex-based differences in age-induced kidney damage.

Introduction: Advanced glycation end products (AGEs) are a heterogeneous group of molecules with potential pathophysiological effects on the kidneys. Fibrosis together with the accumulation of AGEs has been investigated for its contribution to age-related decline in renal function. AGEs mediate their effects in large parts through their interactions with the receptor for AGEs (RAGE). RAGE is a transmembrane protein that belongs to the immunoglobulin superfamily and has the ability to interact with multiple pro-inflammatory/pro-oxidative ligands. The role of RAGE in aging kidneys has not been fully characterized, especially for sex-based differences. Methods: Therefore, we analyzed constitutive RAGE knockout (KO) mice in an age- and sex-dependent manner. Paraffin-embedded kidney sections were used for histological analysis and protein expression of fibrosis and damage markers. RNA expression analysis from the kidney cortex was done by qPCR for AGE receptors, kidney damage, and early inflammation/fibrosis factors. FACS analysis was used for immune cell profiling of the kidneys. Results: Histological analysis revealed enhanced infiltration of immune cells (positive for B220) in aged (>70weeks old) KO mice in both sexes. FACS analysis revealed a similar pattern of enhanced B-1a cells in aged KO mice. There was an age-based increase in pro-fibrotic and pro-inflammatory markers (IL-6, TNF, TGF-β1, and SNAIL1) in KO male mice that presumably contributed to renal fibrosis and renal damage (glomerular and tubular). In fact, in KO mice, there was an age-dependent increase in renal damage (assessed by NGAL and KIM1) that was accompanied by increased fibrosis (assessed by CTGF). This effect was more pronounced in male KO mice than in the female KO mice. In contrast to the KO animals, no significant increase in damage markers was detectable in wild-type animals at the age examined (>70weeks old). Moreover, there is an age-based increase in AGEs and scavenger receptor MSR-A2 in the kidneys. Discussion: Our data suggest that the loss of the clearance receptor RAGE in male animals further accelerates age-dependent renal damage; this could be in part due to an increase in AGEs load during aging and the absence of protective female hormones. By contrast, in females, RAGE expression seems to play only a minor role when compared to tissue pathology.

Towards Non-Invasive Intravital Microscopy: Advantages of Using the Ear Lobe Instead of the Cremaster Muscle.

Inflammation is essential in the protection of the organism and wound repair, but in cases of chronic inflammation can also cause microvasculature deterioration. Thus, inflammation monitorization studies are important to test potential therapeutics. The intravital microscopy (IVM) technique monitors leukocyte trafficking in vivo, being a commonly used procedure to report systemic conditions. Although the cremaster muscle, an established protocol for IVM, may affect the hemodynamics because of its surgical preparation, only male animals are used, and longitudinal studies over time are not feasible. Thinking how this impacts future studies, our aim is to understand if the IVM technique can be successfully performed using the ear lobe instead of the cremaster muscle. Elevated IL-1β plasmatic concentrations confirmed the systemic inflammation developed in a diabetic animal model, while the elevated number of adherent and rolling leukocytes in the ear lobe allowed for the same conclusion. Thus, this study demonstrates that albeit its thickness, the ear lobe protocol for IVM is efficient, non-invasive, more reliable, cost-effective and timesaving.

The Nrf2 Pathway in Depressive Disorders: A Systematic Review of Animal and Human Studies.

There is increasing interest in the involvement of antioxidative systems in protecting from depression. Among these, Nrf2 occupies a central place. We aimed to review the role of Nrf2 in depression. For this reason, we conducted a PubMed search using as search strategy (psychiatr*[ti] OR schizo*[ti] OR psychot*[ti] OR psychos*[ti] OR depress*[ti] OR MDD[ti] OR BD[ti] OR bipolar[ti] OR Anxiety[ti] OR antidepress*[ti] OR panic[ti] OR obsess*[ti] OR compulsio*[ti] OR "mood disord*"[ti] OR phobi*[ti] OR agoraphob*[ti] OR anorex*[ti] OR anorect*[ti] OR bulimi*[ti] OR "eating disorder*"[ti] OR neurodevelopm*[ti] OR retardation[ti] OR autism[ti] OR autistic[ti] OR ASM[ti] OR adhd[ti] OR "attention-deficit"[ti]) AND nrf2, which on the 9th of March produced 208 results of which 89 were eligible for our purposes. Eligible articles were studies reporting data of Nrf2 manipulations or content by any treatment in human patients or animals with any animal model of depression. Most studies were on mice only (N = 58), 20 on rats only, and three on both rats and mice. There were two studies on cell lines (in vitro) and one each on nematodes and fish. Only four studies were conducted in humans, one of which was post mortem. Most studies were conducted on male animals; however, human studies were carried out on both men and women. The results indicate that Nrf2 is lower in depression and that antidepressant methods (drugs or other methods) increase it. Antioxidant systems and plasticity-promoting molecules, such as those in the Nrf2-HO-1, BDNF-TrkB, and cyclic AMP-CREB pathways, could protect from depression, while glycogen synthase kinase-3β and nuclear factor κB oppose these actions, thus increasing depressive-like behaviours. Since Nrf2 is also endowed with tumorigenic and atherogenic potential, the balance between benefits and harms must be taken into account in designing novel drugs aiming at increasing the intracellular content of Nrf2.