Adult Male Research Articles

The Age of Snakes: a comparison of three Canadian species at their northern peripheries

ABSTRACT Aging animals enables an understanding of life-history strategies, population dynamics, and conservation concerns. Such information is particularly wanting for reptiles at their range peripheries where life-history trade-offs make body size an unreliable surrogate for age. Using skeletochronology we quantified the ages of three species sampled near their northern range limits (Common gartersnake – Thamnophis sirtalis, Western Rattlesnake – Crotalus oreganus, Great Basin Gophersnake – Pituophis catenifer). Considerable range in length was seen for snakes of the same age, and the relationship between age and size was stronger in males. Male gartersnakes reached a size associated with sexual maturity in about half the time of females, but for the other two species there was no significant difference between sexes. The oldest snakes sampled were 10–12 years, with one outlier gartersnake estimated at 15 years. Given that females in northern populations cannot reproduce annually, our estimated ages of these snakes suggest reproductive bouts occur only a small number of times during their lifespan, reflecting the precarious existence of northern snake populations. Age data improves our understanding of how life-history varies with locations (including latitude), and further studies that collect contemporary and widespread data on this basic population parameter are required.

In Vitro Assessment of Metarhizium Anisopliae Pathogenicity Against Aedes Aegypti Life Stages.

Aedes aegypti transmits the arboviruses that cause dengue, zika, and chikungunya. Entomopathogenic fungi are beneficial microorganisms that can be incorporated into current strategies against mosquitoes of public health concern. This study molecularly identified the Metarhizium anisopliae CG 153 isolate and evaluated its virulence against larvae, pupae, and adults (both males and females) of Ae. aegypti. Different concentrations of conidia were used (1 × 104-8 conidia mL-1). Larval and pupal survival was monitored daily for seven and three days, respectively, while adults were monitored for 15days. The efficacy of M. anisopliae sensu stricto was concentration-dependent, with higher concentrations achieving better results, demonstrating greater virulence against larval and adult stages of Ae. aegypti. The fungus reduced the larval survival by 95,5% (1 × 108 con.mL-1), 94,4% (1 × 107 con.mL-1), 78,9% (1 × 106 con.mL-1), 62,2% (1 × 105 con.mL-1), and 41,1% (1 × 104 con.mL-1) after seven days. Adults also showed susceptibility to the fungus, with no observed difference in susceptibility between males and females. Over 15days of monitoring, adult survival rates ranged from approximately 6.7% to 72%. Pupae exhibited lower susceptibility to the fungus across different concentrations, with survival rates ranging from approximately 87.8% to 100%. This study highlights the high effectiveness of M. anisopliae CG 153 against both Ae. aegypti larvae and adults (male and female) under controlled conditions, suggesting its promising potential for further evaluation and application in field conditions.

The effect of auriculotherapy on anxiety in older adult candidates for coronary angiography: A randomized controlled trial

AbstractIntroductionCoronary angiography is an effective but anxiety‐provoking procedure in the diagnosis and treatment of many cardiovascular diseases. Anxiety could complicate cardiovascular conditions of older adults and be detrimental to treatment procedures. The current study aimed to examine the effect of auriculotherapy on the anxiety of older adult candidates for coronary angiography.MethodThe study was a randomized controlled trial with a pretest–posttest design. The sample consisted of 94 male and female older adults that were randomly assigned to three groups: the auriculotherapy group with magnetized pellets, the auriculotherapy group with nonmagnetized pellets, and the control group. The collected data were analyzed using analysis of covariance.ResultsThe results showed that auriculotherapy either with magnetized or nonmagnetized pellets would lead to a statistically significant reduction in state and trait anxiety of coronary angiography candidates.ConclusionAuriculotherapy could be considered an adjunct or stand‐alone nonpharmacological method for lowering anxiety before coronary angiography in older adults.

Accounting for sperm whale population demographics in density estimation using passive acoustic monitoring

Sperm whales Physeter macrocephalus are highly sexually dimorphic, with adult males having larger bodies, more powerful echolocation clicks, and slower echolocation clicking rates compared to females. This study introduces methods for estimating sperm whale population densities in the Gulf of Mexico (GoMex) by accounting for the population demographics using passive acoustic monitoring and reveals that ignoring the differences between demographic segments can introduce bias in density estimates. Weekly densities were estimated per 3 demographic segments: social groups consisting of adult females and their offspring, mid-size animals, and adult males. Analysis revealed that the GoMex sperm whale population is primarily composed of social groups, which account for 92 to 98% of the overall population. Mid-size animals and adult males made up a small proportion of the population and were only intermittently present. Our 7 yr GoMex density estimates, including the 2010 Deepwater Horizon (DWH) oil spill period and subsequent years, revealed demographic-specific trends. Declines found at 2 north-central GoMex sites, coupled with increases at a southeastern site, may indicate population movements and potential impacts from the 2010 DWH oil spill and elevated noise levels from anthropogenic activities in the north-central GoMex.

Coverage of community-wide mass drug administration platforms for soil-transmitted helminths in Benin, India, and Malawi: findings from the DeWorm3 project

BackgroundSoil-transmitted helminths (STH) affect approximately 1.5 billion people globally. The current STH control strategy is annual or twice-annual preventive chemotherapy, typically school-based deworming targeting children and women of reproductive age. Mathematical modeling suggests that it may be possible to interrupt STH transmission through high-coverage community-wide mass drug administration (cMDA). DeWorm3 is a cluster randomized trial testing cMDA for prevalence reduction and transmission interruption. The purpose of this study is to describe coverage of cMDA in study clusters over time and correlates of coverage at individual and cluster levels.MethodsFrom 2018–2020, DeWorm3 delivered six rounds of cMDA with 400 mg albendazole at sites in Benin, India, and Malawi. We report coverage, treatment uptake, and directly observed therapy across all rounds. Factors associated with coverage at the cluster level were identified using binomial generalized estimating equations, while factors associated with non-treatment at the individual level were identified using binomial mixed-effects models.ResultsCoverage was high across all clusters and rounds, exceeding the WHO target of 75% in all sites and across all rounds (78% to 95%); cluster-level coverage tended to increase over time. Younger, unmarried, and migratory adults were more likely to be untreated at all sites; adult males were more likely to be untreated in Benin and Malawi. Among children, girls were more likely to be untreated, as were non-school-attending and migratory children. Higher adult education was associated with greater odds of non-treatment among adults, but lower odds among children in the household. Belonging to a less wealthy or minority language-speaking household was associated with non-treatment among both adults and children.ConclusionsIt is possible to deliver community-wide MDA with high coverage. Unique individual and community-level factors influence treatment across settings, and these may be addressed through targeted programming.Trial Registration: Field Studies on the Feasibility of Interrupting the Transmission of Soil-transmitted Helminths (STH), NCT03014167.

Influence of Physical Exercise on the Rehabilitation of Volumetric Muscle Loss Injury Reconstructed with Autologous Adipose Tissue

Background: In volumetric muscle loss (VML) injuries, spontaneous muscle regeneration capacity is limited. The implantation of autologous adipose tissue in the affected area is an option to treat these lesions; however, the effectiveness of this therapy alone is insufficient for a complete recovery of the damaged muscle. This study examined the influence of treadmill exercise on the rehabilitation of VML injuries reconstructed with autologous adipose tissue, as a strategy to counteract the limitations of spontaneous regeneration observed in these injuries. Methods: Forty adult male Wistar rats were divided into eight groups of five individuals each: normal control (NC), regenerative control (RC), VML control (VML), VML injury reconstructed with fresh autologous adipose tissue (FAT), exercise-rehabilitated control (RNC), exercise-rehabilitated regenerative control (RRC), exercise-rehabilitated VML injury (RVML), and exercise-rehabilitated VML injury reconstructed with fresh autologous adipose tissue (RFAT). Histological and histochemical staining techniques were used for the analysis of structural features and histomorphometric parameters of the tibialis anterior muscle. Grip strength tests were conducted to assess muscle force. Results: Exercise rehabilitation decreased the proportion of disoriented fibers in RFAT vs. FAT group. The percentage of fibrosis was significantly higher in FAT and RFAT groups versus NC and RNC groups but did not vary significantly between FAT and RFAT groups. Overall, muscle grip strength and fiber size increased significantly in the exercise-rehabilitated groups compared to control groups. Conclusions: To conclude, rehabilitation with physical exercise tended to normalize the process of muscle repair in a model of VML injury reconstructed with fresh autologous adipose tissue, but it did not reduce the intense fibrosis associated with these injuries.

Evaluation of the therapeutic potential of novel nanoparticle formulations of glutathione and virgin coconut oil in an experimental model of carbon tetrachloride-induced liver failure.

Acute liver failure (ALF) is a critical condition characterized by rapid liver dysfunction, leading to high mortality rates. Current treatments are limited, primarily supportive, and often require liver transplantation. This study investigates the potential of a novel nanoparticle formulation of glutathione (GSH) and virgin coconut oil (VCO) alone and in combination to enhance therapeutic outcomes in a rat model of ALF induced by orogastric carbon tetrachloride (CCl4). The study employed adult male Albino rats divided into ten groups, with ALF induced via a single oral dose of CCl4. Various treatment regimens were administered over seven days, including conventional and nanoparticle forms of GSH and VCO and their combinations. The efficacy of treatments was evaluated through biochemical analysis of liver function markers, oxidative stress indicators, inflammatory biomarkers, and histopathological examinations. Nanoparticles were synthesized using established methods, and characterization techniques were employed to ensure their quality and properties. The nanoparticle formulations significantly improved liver function, as indicated by reduced serum levels of alanine aminotransferase and aspartate aminotransferase, alongside decreased oxidative stress markers such as malondialdehyde. Furthermore, they reduced tumor necrosis factor alpha and interleukin-1 beta inflammatory markers. Histological analysis revealed reduced hepatocellular necrosis and inflammation in treated groups compared to controls. Also, decreased nuclear factor-kappa B was detected by immunohistochemical analysis. The findings show that the nanoparticle mixture of GSH and VCO effectively reduces liver damage in ALF. This suggests a promising drug-based approach for improving liver regeneration and protection. This innovative strategy may pave the way for new therapeutic interventions in the management of ALF.

Половая структура поселений двустворчатого моллюска Macoma calcarea (Gmelin, 1791) в районах с разным гидрологическим режимом

The influence of changes in environmental conditions on the reproductive cycles of benthic organisms (sexual structure, size of puberty, stages of gonad maturity, oocyte size) of the bivalve mollusk Macoma calcarea in the western part of the Spitsbergen archipelago (Grenfjord Bay) and the eastern part of the Barents Sea (coastal archipelago) was studied New Earth). In a settlement of M. calcarea in the warmer waters of Grenfjord, intensive reproductive processes were observed. In this area, females reach sexual maturity with a shorter shell length than males and individuals from other areas of the Barents Sea; In terms of sex ratio, females dominate over males, and a large number of juveniles are noted. Females at the spawning stage were found only near Novaya Zemlya. The diameter of the oocytes varied from 30 to 200 µm depending on the stage of maturity. In the mollusks M. calcarea, the breeding season is extended in time in both study areas.

Effect of climatic factors on sexual maturity Labeo rohita fish

<p>This study examined the varying levels of thyroxine hormone impact (at doses of 2, 10, and 50 µg g-1 BW) as an environmentally responsive hormone on the fatty acid and sexual steroid profiles in male Labeo rohita fish. Each treatment group comprised six male fish with an average weight of 5.4 ± 0.58 kg. Blood samples were collected from male rohu fish pre-and post-injection, as well as following sperm retrieval from the caudal peduncle. Biotechnical parameters were evaluated post-fertilization. Among male breeders, the pituitary treatment and the 50 µg g-1 BW thyroxine treatment exhibited the highest levels of estradiol, testosterone, 17-alpha hydroxyprogesterone, cortisol, LH, as well as testicular protein and fat content. Whereas the lowest levels were observed in the LHRH recipient and control groups (p < 0.05). Moreover, the saturated fatty acids C16:0 and C18:0, monounsaturated fatty acids C18:1n9c and C24:1n9, along with polyunsaturated fatty acids C20:3n6 and C22:6n3 in male breeders, were found to be the predominant fatty acids in the testes under hormonal treatment. The pituitary treatment and the 50 µg g-1 BW thyroxine treatment demonstrated a greater total percentage of testicular fatty acids compared to other interventions (p < 0.05). These findings emphasize the heightened efficacy of pituitary hormone and subsequent thyroxine hormone administration at a dose of 50 µg g-1 BW, relative to lower thyroxine doses and LHRH. The aforementioned treatment can increase the levels of sex hormones that are important for sexual responsiveness, emphasizing their pivotal role in facilitating successful artificial reproduction in male Rohu breeders.</p>

“Close-loop Urethral Stricture:" A Variant of Multiple Urethral Stricture Disease - Its Description, Management and Outcome: A Case Report

Abstract Urethral stricture disease is an abnormal narrowing and loss of distensibility of the urethra due to spongiofibrosis. It is commoner in young adult males. The management of this condition is diverse with various factors influencing the choice of treatment. However, length of the stricture is a major determinant of the choice of treatment. Two or more stricture sites close together are also treated as long segment stricture. Our aim is to describe a variant of multiple urethral strictures consisting of two short segment complete strictures involving the proximal bulbar urethral and urethral meatus with intervening significant length of normal urethra "close-loop stricture" in a 46-year old man who had traumatic urethral catheterization and was referred to the urology clinic six months following suprapubic tube cystostomy. KeywordsStricture, Meatotomy, Urethroplasty, Catheterization, Bouginage

Comparative Analysis of Verapamil Pharmacokinetics: Evaluating the Impact of Simple Suspension and Crushing Administration Methods

Background/Objective: It is not uncommon for elderly patients to experience difficulties with feeding and swallowing. In the simple suspension method, tablets are dissolved and suspended in warm water without prior crushing or decapsulation, and then administered via a tube. Despite the prevalence of this method, the pharmacokinetics of suspended tablet dosage forms remain poorly understood. Methods: Verapamil was employed in dissolution tests following both the simple suspension and crushing methods. A pharmacokinetics study was conducted on healthy adult males. Results: The resultant dissolution profiles from the two methods exhibited notable dissimilarities. Drug release from the crushed product commenced earlier than that from the simple suspension and intact tablet. Furthermore, the area under the curve for verapamil during the initial 24 h period was 1.7 and 1.3 times greater in the crushed and simple suspension groups, respectively, than in the tablet group. Conclusions: The crushing and simple suspension methods are safe techniques for administering medications to patients with dysphagia, thereby preventing aspiration. Nevertheless, the processing of medications may result in an increased frequency of adverse effects. It is recommended that the processing of medicines prior to administration be avoided.

Working memory performance predicts, but does not reduce, cocaine- and cannabinoid-seeking in adult male rats.

Cognitive deficits reflecting impaired executive function are commonly associated with psychiatric disorders, including substance use. Cognitive training is proposed to improve treatment outcomes for these disorders by promoting neuroplasticity within the prefrontal cortex, enhancing executive control, and mitigating cognitive decline due to drug use. Additionally, brain derived neurotrophic factor (BDNF) can facilitate plasticity in the prefrontal cortex and reduce drug-seeking behaviors. We investigated whether working memory training could elevate BDNF levels in the prefrontal cortex and if this training would predict or protect against cocaine or cannabinoid seeking. Adult male rats were trained to perform a 'simple' or 'complex' version of a delayed-match-to-sample working memory task. Rats then self-administered cocaine or the synthetic cannabinoid WIN55,212-2 and were tested for cued drug-seeking during abstinence. Tissue from the prefrontal cortex and dorsal hippocampus was analyzed for BDNF protein expression. Training on the working memory task enhanced endogenous BDNF protein levels in the prelimbic prefrontal cortex but not the dorsal hippocampus. Working memory training did not impact self-administration of either drug but predicted the extent of WIN self-administration and cocaine seeking during abstinence. These results suggest that working memory training promotes endogenous BDNF but does not alter drug-seeking or drug-taking behavior. However, individual differences in cognitive performance prior to drug exposure may predict vulnerability to future drug use.

The diet and diving behaviour of the ringed seal (Pusa hispida) in Kangia (Ilulissat Icefjord), Greenland

AbstractThis study uses telemetry and stomach content analysis to describe the ringed seals’ dive pattern and prey selection within Kangia (Ilulissat Icefjord), Greenland, where the ringed seals are known to be exceptionally large and genetically distinct. Data from twenty-four ringed seals (Pusa hispida) tagged in Kangia, with satellite relay data loggers from 2012 to 2020, were used to describe the dive patterns. The transmitters were active between 28 and 270 days and showed that subadults most frequently dove to around 150 m. Adult males frequently dove to this depth, but they also had a second frequently visited depth at around 400 m, some adults significantly deeper. Adult males generally dove deeper than females and adult seals showed deeper dives during the winter, a feature that was less prominent in subadults. The stomach content from ringed seals (n = 108) harvested in Kangia by subsistence hunters revealed that polar cod (Boreogadus saida), capelin (Mallotus villosus), mysids (Mysis oculata, M. littoralis, M. Polaris) and shrimps (mainly Pandalus borealis) were the dominant prey. Shrimps were observed more often in adults, likely due to the frequently visited dive depth of around 400 m, whereas mysids were more observed in small subadult seals. The stomach content from harp seals (Pagophilus groenlandicus) and Greenland halibut (Reinhardtius hippoglossoides) caught in Kangia also showed a strong niche overlap with the ringed seals. These findings outline the unique ecological adaptations and the importance of Kangia as a critical habitat of ringed seals.

Biological Manifestation of Different Temperature Regimes and Certain Insecticides on Cheilomenes sexmaculata (Fab.) Preying on Cowpea Aphid, Aphis craccivora Koch

Background: Ladybird beetles are great bio-control agents and campodeiform grubs of these little predators are able to devour aphids in a voracious fashion. They feed on a wide array of aphid species most noticeably on the infamous black aphids or cowpea aphids Aphis craccivora (Koch). Methods: The growth and feeding potential of C. sexmaculata were studied in the laboratory of the Department of Entomology, Faculty of Agricultural Sciences (Institute of Agricultural Sciences), SOA Deemed to be University, Bhubaneswar, India at four different temperatures 20°C, 25°C, 30°C and 35°C during 2021-2022. Result: The life history parameters indicated that the duration of eggs, larvae, pupae, adult males and adult females was longest at 30°C and shortest at 20°C. Such variations were also observed in morphometry and feeding potential. The trend was 30°C greater than 25°C greater than 35°C greater than 20°C. In the case of feeding potential the trend was adult female greater than adult male greater than 4th instar greater than 3rd instar greater than 2nd instar greater than 1st instar across all temperatures. Adult females lived longer than males and thereby consumed more aphids than males. Maximum consumption was seen in case of adult female at 30°C which was 633±5.71 aphids and least feeding was seen at 20°C by 1st instar grub which was 8.9±0.56 aphids. The effect of insecticides in direct spray method indicated that dimethoate 30EC killed more ladybirds and clothianidin 50% WDG killed the least. This trend was strong across all temperatures and insect growth stages. The feed contamination method indicated that clothianidin 50% WDG, imidacloprid 17.8% SL and thiamethoxam 25% WG produced higher mortality rates than direct contamination method. All insecticides showed increased mortality up to 72 h and the mortality trend was 72 HAT greater than 48 HAT greater than 24 HAT. The mortality rate of immature individuals is higher than that of adults.

Effects of Vitamin C on Aluminum Chloride- Induced Neurotoxicity on the Hippocampal Cortex of Adult Wistar Male Rats

This study aimed to evaluate the neuroprotective effects of Vitamin C on aluminum chloride (AlCl₃)- induced brain damage, focusing on behavioral, biochemical, and histomorphological outcomes in a rodent model. Study Design: A total of 42 healthy male rats were randomly assigned to three groups: control, AlCl₃ -only, and AlCl₃ plus Vitamin C (500 mg/kg and 1000 mg/kg). The AlCl₃ groups received daily doses of AlCl₃, while the Vitamin C groups received concurrent Vitamin C supplementation. Methodology: Body weight, behavioral changes, biochemical markers of oxidative stress (MDA, SOD, GSH), and inflammation (TNF-α, IL-6) were assessed. Histomorphological analysis of the hippocampus was performed to evaluate structural damage. Behavioral assessments included locomotor and exploratory activity tests. Biochemical analyses measured oxidative stress and antioxidant enzyme activities. Results: AlCl₃ administration resulted in significant body weight loss, increased oxidative stress (elevated MDA, and SOD levels), and heightened inflammation (elevated TNF-α and IL-6). Behavioral tests showed increased excitatory activities, and increased anxiety levels as seen in increased rearing and grooming activities. Histomorphological examination revealed significant hippocampal damages as seen in the pale staining, shrunken pyramidal cells. Vitamin C co- administration mitigated these adverse effects, demonstrating reduced body weight loss, lower oxidative stress, decreased inflammation, and improved behavioral performance. Histological analysis indicated less hippocampal damage in the Vitamin C treated groups. Conclusion: As an antioxidant, Vitamin C effectively attenuates the neurotoxic effects of AlCl₃ by reducing oxidative stress and inflammation, and by protecting the hippocampal cellular integrity. These findings suggest that Vitamin C holds potential as a therapeutic agent for mitigating neurotoxicity induced by aluminum compounds.

Oral pre-treatment with Citronellol ameliorates Methotrexate-induced nephrotoxicity in Wistar rats via targeting oxidative stress and inflammation

Background Methotrexate (MTX) is a classical folic acid antagonist widely used in the treatment of malignant and non-malignant disorders. However, its clinical application is often restricted by concomitant adverse effects, including renal damage. Numerous studies have highlighted the role of oxidative stress and inflammation in mediating MTX-related nephrotoxicity. Therefore, the current study aimed to explore the possible renoprotective action of Citronellol (CT), a natural compound with prominent antioxidant and anti-inflammatory activities, against nephrotoxicity induced by MTX. Methods To fulfill our objective, 24 adult male Wistar rats were randomly allocated into four groups: control, MTX, 100 mg/kg CT plus MTX and 200 mg/kg CT plus MTX. At the end of the study, the experimental rats were anesthetized, blood samples were collected for biochemical assays, and the kidneys were surgically removed for biochemical and gene expression analyses, after which all rats were sacrificed by exsanguination. Results Compared to the MTX-treated group, our results revealed that pre-supplementation with 100 or 200 mg/kg CT remarkably ameliorated renal damage biomarkers, including serum urea, serum creatinine, and kidney injury molecule-1 (KIM-1). In addition, pre-treatment with 100 or 200 mg/kg CT enhanced the activity of the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx), diminished renal malondialdehyde (MDA) contents, and attenuated inflammation by suppressing renal nuclear factor kappa B (NF-κB) signaling and diminishing tumor necrosis factor-alpha (TNF-α) gene expression. Moreover, pre-treatment with 200 mg/kg CT markedly reduced interleukin-1 beta (IL-1β) gene expression. Conclusion Our findings demonstrate, for the first time, that CT can serve as a new promising agent for mitigating nephrotoxicity induced by MTX through its antioxidant and anti-inflammatory properties.

Combined Method Comprising Low Burden Physiological Measurements with Dry Electrodes and Machine Learning for Classification of Visually Induced Motion Sickness in Remote-Controlled Excavator

The construction industry is actively developing remote-controlled excavators to address labor shortages and improve work safety. However, visually induced motion sickness (VIMS) remains a concern in the remote operation of construction machinery. To predict the occurrence and severity of VIMS, we developed a prototype system that acquires multiple physiological signals with different mechanisms under a low burden and detects VIMS from the collected data. Signals during VIMS were recorded from nine healthy adult males operating excavator simulators equipped with multiple displays and a head-mounted display. Light gradient-boosting machine-based VIMS detection binary classification models were constructed using approximately 30,000 s of time-series data, comprising 23 features derived from the physiological signals. These models were validated using leave-one-out cross-validation on seven participants who experienced severe VIMS and evaluated through area under the curve (AUC) scores. The mean receiver operating characteristic curve AUC score was 0.84, and the mean precision–recall curve AUC score was 0.71. All features were incorporated into the models, with saccade frequency and skin conductance response identified as particularly important. These trends aligned with subjective assessments of VIMS severity. This study contributes to advancing the use of remote-controlled machinery by addressing a critical challenge to operator performance and safety.

8509 Molecular and Reproductive Assessment in Ames strain under or not Hormonal Replacement

Abstract Disclosure: B.V. Azevedo: None. N. Trigueiro: None. J.M. Marques: None. V. Yariwake: None. L. Tamashiro: None. R. Cruz: None. M.M. Veras: None. L. Carvalho: None. Introduction: The Ames lineage, a mouse model with a mutation in the Prop1 gene, exhibits GH, TSH, and PRL deficiencies along with low levels of gonadotropins. This has prompted an investigation into aging and reproductive restoration after hormone replacement reported in the literature. Aims: The present study aims to analyze the pathophysiologic mechanisms in the reproductive system and fertility restoration. Methods: This is a prospective study approved by the ethical committee for animal use at the University of Sao Paulo Medical School. An isogenic Ames lineage was categorized into four groups: 1) Ames wild-type mice, 2) untreated Ames dwarf mice with sexual maturity, 3) untreated Ames dwarf mice without sexual maturity, and 4) Ames dwarf mice under hormonal treatment. Male Ames dwarf mice received treatment with levothyroxine and human recombinant GH, starting at 30 days postnatal (dp) for 40 days, followed by weekly applications until 90 dp. At the time of euthanasia, a set of analyses were conducted on animals from each group to characterize sexual maturation, fertility, and tissue collection for reproductive parameter analyses, such as seminiferous tubule morphology by histology, testicular weight, gonadosomatic index, and spermogram analysis. Additionally, pituitary RNAseq was performed using the poly-A tail capture method with the Illumina stranded mRNA prep kit and sequenced on the NextSeq 2000. Results: Treated dwarf mice exhibited fertility restoration, an increase in testicular volume with an improvement of testicular function, and complete spermatogenesis. Untreated Ames dwarf mice were infertile, but those that reached sexual maturity without hormonal replacement showed improvements in reproductive parameters, such as enhanced testicular development, maturation of some seminiferous tubules, increased testicular weight, and improvement in the spermogram compared to immature Ames dwarf mice. Transcriptome analyses revealed distinct patterns of all Ames dwarf mice groups and the wild-type. It was observed a decrease in Fshb expression in the group without sexual maturation compared to the wild-type group. Additionally, Prl and Slc6a12 were upregulated in the spontaneously sexually mature Ames dwarf compared to the non-restored Ames dwarf mice. Conclusion: GH and levothyroxine treatment in the Ames lineage are effective in restoring fertility and improving reproductive parameters in male Ames dwarf mice. However, sexually mature Ames dwarf mice without treatment presented improvements in reproductive and molecular parameters despite remaining infertile. These findings provide valuable insights into the complex interaction among hormonal replacement, modulating mechanisms, and sexual maturity. Presentation: 6/2/2024

8509 Molecular and Reproductive Assessment in Ames strain under or not Hormonal Replacement

Abstract Disclosure: B.V. Azevedo: None. N. Trigueiro: None. J.M. Marques: None. V. Yariwake: None. L. Tamashiro: None. R. Cruz: None. M.M. Veras: None. L. Carvalho: None. Introduction: The Ames lineage, a mouse model with a mutation in the Prop1 gene, exhibits GH, TSH, and PRL deficiencies along with low levels of gonadotropins. This has prompted an investigation into aging and reproductive restoration after hormone replacement reported in the literature. Aims: The present study aims to analyze the pathophysiologic mechanisms in the reproductive system and fertility restoration. Methods: This is a prospective study approved by the ethical committee for animal use at the University of Sao Paulo Medical School. An isogenic Ames lineage was categorized into four groups: 1) Ames wild-type mice, 2) untreated Ames dwarf mice with sexual maturity, 3) untreated Ames dwarf mice without sexual maturity, and 4) Ames dwarf mice under hormonal treatment. Male Ames dwarf mice received treatment with levothyroxine and human recombinant GH, starting at 30 days postnatal (dp) for 40 days, followed by weekly applications until 90 dp. At the time of euthanasia, a set of analyses were conducted on animals from each group to characterize sexual maturation, fertility, and tissue collection for reproductive parameter analyses, such as seminiferous tubule morphology by histology, testicular weight, gonadosomatic index, and spermogram analysis. Additionally, pituitary RNAseq was performed using the poly-A tail capture method with the Illumina stranded mRNA prep kit and sequenced on the NextSeq 2000. Results: Treated dwarf mice exhibited fertility restoration, an increase in testicular volume with an improvement of testicular function, and complete spermatogenesis. Untreated Ames dwarf mice were infertile, but those that reached sexual maturity without hormonal replacement showed improvements in reproductive parameters, such as enhanced testicular development, maturation of some seminiferous tubules, increased testicular weight, and improvement in the spermogram compared to immature Ames dwarf mice. Transcriptome analyses revealed distinct patterns of all Ames dwarf mice groups and the wild-type. It was observed a decrease in Fshb expression in the group without sexual maturation compared to the wild-type group. Additionally, Prl and Slc6a12 were upregulated in the spontaneously sexually mature Ames dwarf compared to the non-restored Ames dwarf mice. Conclusion: GH and levothyroxine treatment in the Ames lineage are effective in restoring fertility and improving reproductive parameters in male Ames dwarf mice. However, sexually mature Ames dwarf mice without treatment presented improvements in reproductive and molecular parameters despite remaining infertile. These findings provide valuable insights into the complex interaction among hormonal replacement, modulating mechanisms, and sexual maturity. Presentation: 6/2/2024

Astrocytes contribute to the functional differentiation of the hippocampal longitudinal axis during reward and aversion processing in the adult male rat

This study aims to investigate whether glial cells, in particular putative astrocytes, contribute to functional distinctions between the dorsal (DH), intermediate (IH), and ventral (VH) hippocampus. To evaluate this, we performed three different behavioral tasks (i.e., Morris water maze; MWM, Passive avoidance; PA, T-maze place preference; TPP) to determine whether the DH, IH, and VH are necessary for each task. Sensitivity of behavioral tasks was confirmed using lidocaine (2 %, 1 μl) reversible inactivation. Subsequently, we examined the effects of silencing astrocytes, using fluorocitrate (FC, 1 mM/1 μl), into the DH, IH, and VH on these tasks. The effects of drugs were examined separately. We observed that injection of FC into the DH resulted in a significant impairment in MWM performance. In contrast, while FC injections into the IH or VH did not prevent platform localization during the acquisition phase, rats showed difficulty recalling the target zone during the retrieval phase. In the PA test, FC injection into the VH impaired task learning and memory. During the acquisition phase, FC injection into the DH or IH did not differ from the control in the number of shocks; however, during retrieval, there was a significant decrease in the latency before entering the dark chamber. The TPP test performance was impaired by FC injection in the IH. In sum, we show that glial cells, especially astrocytes in specific functional regions of the hippocampus, play distinct roles in processing aversive and rewarding experiences and contribute to the functional organization of the hippocampal longitudinal axis.