Urogenital Tissues Research Articles

Bacteriophage P2-71: a promising therapeutic against multidrug-resistant Proteus mirabilis in urinary tract infections.

Proteus mirabilis is a Gram-negative, rod-shaped bacterium widely found in natural environments. It is known for causing a range of severe illnesses in mammals, particularly urinary tract infections (UTIs). This study evaluates the therapeutic efficacy of phage P2-71 against Proteus mirabilis in vivo and in vitro environments. The in vitro therapeutic potential of bacteriophage P2-71 was assessed through the ability of phage to kill Proteus mirabilis by using a plate counting assay, and biofilm inhibition and biofilm lysis assays using a microtitre plate method. Additionally, an in vivo UTI model in C57BL/6Jmice was developed via urethral inoculation of the bacterium. Phage therapy was administered through urethral injection over a period of 5 days. Therapeutic outcomes were measured by analyzing bacterial load, phage titer, inflammatory markers, and histopathological changes in the urine, urogenital tissues, and spleen. In vitro, bacteriophage P2-71 achieved significant reductions in P. mirabilis concentrations, with log reductions of 1.537 and 0.7009 CFU/mL in laboratory and urine environments, respectively (p < 0.001). The phage also decreased biofilm formation by 34-49% and lysed 15-25% of mature biofilms at various multiplicities of infection (MOIs) (p < 0.001). In vivo, phage treatment significantly lowered bacterial concentrations in the urine on Days 1 and 3 (p < 0.0001), achieving a maximum reduction of 4.602 log₁₀ CFU/mL; however, its effectiveness diminished by Day 5 (p > 0.05). Concurrently, phage titers decreased over time. Importantly, phage treatment notably reduced bacterial load in the bladder, kidneys, and spleen (p < 0.001). Inflammatory markers such as IL-6, IL-1β, and TNF-α were significantly lower in the treatment group, especially in the bladder (p < 0.0001), indicating an effective reduction in inflammation. Histopathological analysis showed significant mitigation of tissue damage. The results demonstrated that bacteriophage P2-71 is a promising alternative therapy for UTIs caused by MDR Proteus mirabilis. This bacteriophage therapy offers a viable strategy for managing infections where traditional antimicrobials fail, highlighting its potential in clinical applications.

Chlamydia muridarum Associated Pulmonary and Urogenital Disease and Pathology in a Colony of Enzootically Infected Il12rb2 Deficient and Stat1 Knockout Mice.

Chlamydia muridarum (Cm), an intracellular bacterium of historical importance, was recently rediscovered as moderately prevalent in research mouse colonies. Cm was first reported as a causative agent of severe pneumonia in mice about 80 y ago, and while it has been used experimentally to model Chlamydia trachomatis infection of humans, there have been no further reports of clinical disease associated with natural infection. We observed clinical disease and pathology in 2 genetically engi- neered mouse (GEM) strains, Il12rb2 KO and STAT1 KO, with impaired interferon-γ signaling and Th1 CD4+ T cell responses in a colony of various GEM strains known to be colonized with and shedding Cm. Clinical signs included poor condition, hunched posture, and poor fecundity. Histopathology revealed disseminated Cm with lesions in pulmonary, gastrointestinal, and urogenital tissues. The presence of Cm was confirmed using both immunohistochemistry for Cm major outer membrane protein-1 antigen and in situ hybridization using a target probe directed against select regions of Cm strain Nigg. Cm was also found in association with a urothelial papilloma in one mouse. These cases provide additional support for excluding Cm from research mouse colonies.

Single-dose Tadalafil Reduces Opening Urethral Pressure: A Randomized, Double-blind, Placebo-controlled, Crossover Trial in Healthy Women

Introduction and HypothesisPhosphodiesterase enzymes are widely distributed in female urogenital tissues. Yet, the understanding of their physiological roles and the impact of phosphodiesterase inhibitors on lower urinary tract symptoms in women remains limited. Current hypotheses are conflicting: one suggests that vasodilation might expand the periurethral vascular plexus, leading to increased urethral pressure, whereas the other proposes a relaxation of urethral musculature, resulting in decreased pressure. To further clarify this, we investigated the effect of tadalafil on the opening urethral pressure and voiding function in healthy women.MethodsWe conducted a randomized, double-blind, placebo-controlled crossover trial involving 24 healthy women. Participants were randomly assigned to receive a single dose of tadalafil (40 mg) or placebo during their initial visit and then switched to the alternative treatment during their second visit. Opening urethral pressure was measured with urethral pressure reflectometry during both resting and squeezing conditions of the pelvic floor. Subsequently, voiding parameters were recorded.ResultsCompared with placebo, a single dose of tadalafil significantly reduced opening urethral pressure during both resting (-6.8 cmH20; 95% confidence interval [CI], -11.8 to -1.9; p = 0.009) and squeezing conditions (-8.8 cmH20; 95% CI, -14.6 to -3.1; p = 0.005). Voiding parameters did not show significant differences (average flow rate: -0.8 ml/s [95% CI, -2.0 to 0.4; p = 0.2]; maximum flow rate: -1.7 ml/s [95% CI, -4.8 to 1.5; p = 0.3]).ConclusionsA single dose of 40 mg tadalafil moderately reduced urethral pressure in healthy women, without affecting voiding parameters. The clinical implications of this are yet to be determined.

Comparative pathogenicity of infectious bronchitis virus Massachusetts and Delmarva (DMV/1639) genotypes in laying hens.

Infectious bronchitis (IB) is a highly contagious and acute viral disease of chicken caused by the infectious bronchitis virus (IBV) of the family Coronaviridae. Even with extensive vaccination against IB by the poultry industry, the occurrence of new IBV genotypes is a continuous challenge encountered by the global poultry industry. This experiment was designed to compare the pathogenicity of two IBV strains belonging to Massachusetts (Mass) and Delmarva DMV/1639 genotypes. Specific pathogen-free laying hens were challenged during the peak of production (30 weeks), keeping a mock-infected control group. During 21 days of observation following infection, a significant drop in egg production with miss-shaped and soft shells was observed in the DMV/1639 IBV-infected hens only. The DMV/1639 IBV infected group showed prolonged and higher cloacal viral shedding compared with the Mass IBV-infected group. At the end of the study (21 days post-infection), the viral genome loads in the respiratory, urogenital, and immune tissues were significantly higher in the DMV/1639 IBV-infected group compared with the Mass IBV-infected group. Macroscopic lesions such as distorted ova leading to egg peritonitis were observed only in the DMV/1639 IBV-infected group. Moreover, microscopic lesion scores were significantly higher in the lung, kidney, cecal tonsils, and oviduct of the DMV/1639 IBV-infected group compared with the Mass IBV-infected group. Finally, the apoptosis index in the kidney, ovary, magnum, isthmus, and shell gland was significantly higher in the DMV/1639 IBV-infected group compared with the control and Mass-infected groups. This study examined the pathogenicity of two IBV genotypes that are impacting the layer industry in North America.

Chlamydia muridarum Associated Pulmonary and Urogenital Disease and Pathology in a Colony of Enzootically Infected Il12rb2 Deficient and Stat1 Knockout Mice

Chlamydia muridarum (Cm), an intracellular bacterium of historical importance, was recently rediscovered as moderately prevalent in research mouse colonies. Cm was first reported as a causative agent of severe pneumonia in mice about 80 y ago, and while it has been used experimentally to model Chlamydia trachomatis infection of humans, there have been no further reports of clinical disease associated with natural infection. We observed clinical disease and pathology in 2 genetically engineered mouse (GEM) strains, Il12rb2KO and STAT1KO, with impaired interferon-γ signaling and Th1 CD4+ T cell responses in a colony of various GEM strains known to be colonized with and shedding Cm. Clinical signs included poor condition, hunched posture, and poor fecundity. Histopathology revealed disseminated Cm with lesions in pulmonary, gastrointestinal, and urogenital tissues. The presence of Cm was confirmed using both immunohistochemistry for Cm major outer membrane protein-1 antigen and in situ hybridization using a target probe directed against select regions of Cm strain Nigg. Cm was also found in association with a urothelial papilloma in one mouse. These cases provide additional support for excluding Cm from research mouse colonies.

Hormonal Medications for Genitourinary Syndrome of Menopause.

Genitourinary syndrome of menopause is a common, under-reported, and undertreated chronic progressive condition requiring long-term treatment. Hypoestrogenism in the urogenital tissues is associated with bothersome dyspareunia, vulvovaginal symptoms, overactive bladder, and frequent urinary tract infections. Vaginal hormone therapies, including vaginal estrogen and intravaginal dehydroepiandrostenedione, are safe and effective and improve symptoms and clinical findings. Systemic hormone therapy treats vulvovaginal atrophy less effectively than vaginal hormone therapies with increased stress and urge urinary incontinence. Oral ospemifene effectively treats vaginal dryness and dyspareunia. Clinicians need to ask about symptoms of genitourinary syndrome of menopause, confirm the diagnosis, and suggest appropriate treatment options.

Induction of kidney-related gene programs through co-option of SALL1 in mole ovotestes.

Changes in gene expression represent an important source of phenotypic innovation. Yet how such changes emerge and impact the evolution of traits remains elusive. Here, we explore the molecular mechanisms associated with the development of masculinizing ovotestes in female moles. By performing integrative analyses of epigenetic and transcriptional data in mole and mouse, we identified the co-option of SALL1 expression in mole ovotestes formation. Chromosome conformation capture analyses highlight a striking conservation of the 3D organization at the SALL1 locus, but an evolutionary divergence of enhancer activity. Interspecies reporter assays support the capability of mole-specific enhancers to activate transcription in urogenital tissues. Through overexpression experiments in transgenic mice, we further demonstrate the capability of SALL1 to induce kidney-related gene programs, which are a signature of mole ovotestes. Our results highlight the co-option of gene expression, through changes in enhancer activity, as a plausible mechanism for the evolution of traits.

Highly multiplexed immune profiling throughout adulthood reveals kinetics of lymphocyte infiltration in the aging mouse prostate.

Aging is a significant risk factor for disease in several tissues, including the prostate. Defining the kinetics of age-related changes in these tissues is critical for identifying regulators of aging and evaluating interventions to slow the aging process and reduce disease risk. An altered immune microenvironment is characteristic of prostatic aging in mice, but whether features of aging in the prostate emerge predominantly in old age or earlier in adulthood has not previously been established. Using highly multiplexed immune profiling and time-course analysis, we tracked the abundance of 29 immune cell clusters in the aging mouse prostate. Early in adulthood, myeloid cells comprise the vast majority of immune cells in the 3-month-old mouse prostate. Between 6 and 12 months of age, there is a profound shift towards a T and B lymphocyte-dominant mouse prostate immune microenvironment. Comparing the prostate to other urogenital tissues, we found similar features of age-related inflammation in the mouse bladder but not the kidney. In summary, our study offers new insight into the kinetics of prostatic inflammaging and the window when interventions to slow down age-related changes may be most effective.

Pharmacodynamic evaluation of lefamulin in the treatment of gonorrhea using a hollow fiber infection model simulating Neisseria gonorrhoeae infections.

The emergence and spread of antimicrobial resistance in Neisseria gonorrhoeae is seriously threatening the treatment and control of gonorrhea globally. Novel treatment options are essential, coupled with appropriate methods to pharmacodynamically examine the efficacy and resistance emergence of these novel drugs. Herein, we used our dynamic in vitro hollow fiber infection model (HFIM) to evaluate protein-unbound lefamulin, a semisynthetic pleuromutilin, against N. gonorrhoeae. Dose-range and dose-fractionation experiments with N. gonorrhoeae reference strains: WHO F (susceptible to all relevant antimicrobials), WHO X (extensively drug-resistant, including ceftriaxone resistance), and WHO V (high-level azithromycin resistant, and highest gonococcal MIC of lefamulin (2mg/l) reported), were performed to examine lefamulin gonococcal killing and resistance development during treatment. The dose-range experiments, simulating a single oral dose of lefamulin based on human plasma concentrations, indicated that ≥1.2g, ≥2.8g, and ≥9.6g of lefamulin were required to eradicate WHO F, X, and V, respectively. Dose-fractionation experiments, based on human lefamulin plasma concentrations, showed that WHO X was eradicated with ≥2.8g per day when administered as q12h (1.4g twice a day) and with ≥3.6g per day when administered as q8 h (1.2g thrice a day), both for 7days. However, when simulating the treatment with 5-10 times higher concentrations of free lefamulin in relevant gonorrhea tissues (based on urogenital tissues in a rat model), 600mg every 12h for 5days (approved oral treatment for community-acquired bacterial pneumonia) eradicated all strains, and no lefamulin resistance emerged in the successful treatment arms. In many arms failing single or multiple dose treatments for WHO X, lefamulin-resistant mutants (MIC = 2mg/l), containing an A132V amino acid substitution in ribosomal protein L3, were selected. Nevertheless, these lefamulin-resistant mutants demonstrated an impaired biofitness. In conclusion, a clinical study is warranted to elucidate the clinical potential of lefamulin as a treatment option for uncomplicated gonorrhea (as well as several other bacterial STIs).

Menopause and genitourinary health: optimal support and correction of disorders

The vulva, vagina, lower urinary tract, and pelvic floor have the same embryonic origin. Thus, all of the abovementioned organs are equally sensitive to estrogen deficiency and menopausal hormone therapy due to expression of the respective receptors. Genitourinary syndrome of menopause (GSM) is a concept first introduced in 2013 to provide a more complete definition of the consequences of estrogen deficiency in urogenital tissues and reflect the full range of vaginal and urinary symptoms that women experience during menopause. The term broadly reflects the condition, which, unlike the older term “vulvovaginal atrophy”, is not limited to a single symptom of dyspareunia and includes women who are not sexually active. The consequences of GSM significantly alter the quality of life of most menopausal women. Some patients with symptoms of vulvovaginal atrophy completely stop their sexual activity (58 %) and avoid sexual intimacy (55%). Various forms of urinary disorders also alter activities of daily living, sleep, sexual activity, and can lead to social isolation and loss of self-respect. Women with recurrent vulvovaginal infection often have to seek medical care. Estrogen therapy during menopause is pathogenetically justified. Estrogens administered intravaginally are more effective in relieving the symptoms of GSM, both objectively and subjectively, than their systemic use. Up to 40% of women receiving systemic menopausal hormone therapy do not have the expected effect of estrogens on the urogenital tract. If the symptoms of GSM prevail, it is advisable to consider monotherapy with local estrogens or combine it with systemic therapy instead of increasing a dose of the latter. The article presents a clinical case of a personalized approach to solving the issue of vulvovaginal atrophy combined with a mixed urinary incontinence (MUI) in a postmenopausal female patient.

Tissue Distribution of [14C]-Lefamulin into the Urogenital Tract in Rats.

ABSTRACTLefamulin, a semisynthetic pleuromutilin antibiotic approved in the United States, Canada, and Europe for intravenous and oral treatment of community-acquired bacterial pneumonia, is highly active in vitro against bacterial pathogens that cause sexually transmitted infections (STIs), including multidrug-resistant strains of Neisseria gonorrhoeae, Chlamydia trachomatis, and Mycoplasma genitalium. This nonclinical study used quantitative whole-body autoradiography (QWBA) and qualitative tape-transfer microautoradiography (MARG) to investigate lefamulin distribution into urogenital tract tissues down to a cellular level in male and female rats. A single intravenous dose (30 mg/kg) of [14C]-lefamulin was administered to 3 male and 3 female Sprague-Dawley rats. At 0.5, 6, and 24 h post dose, rats were euthanized and [14C]-lefamulin distribution was investigated using QWBA and MARG of sagittal planes. [14C]-lefamulin was well distributed throughout the carcasses of male and female rats, with the highest concentrations observed in male bulbourethral gland, urethra, prostate in female clitoral gland, uterus (particularly endometrium), and ovary. In these areas, concentrations were similar to or higher than those observed in the lungs. Concentrations peaked at 0.5 h post dose, remaining detectable in the urogenital tract up to 24 h post dose. [14C]-lefamulin in rats showed rapid, homogeneous distribution into urogenital tissues down to a cellular level, with high tissue:blood ratios in tissues relevant to STI treatment. These results, and the potent in vitro activity of lefamulin against multidrug-resistant bacteria known to cause STIs, will help inform further assessment of lefamulin, including potential clinical evaluation for treatment of STIs.

Chromosome-level genome of Schistosoma haematobium underpins genome-wide explorations of molecular variation.

Urogenital schistosomiasis is caused by the blood fluke Schistosoma haematobium and is one of the most neglected tropical diseases worldwide, afflicting > 100 million people. It is characterised by granulomata, fibrosis and calcification in urogenital tissues, and can lead to increased susceptibility to HIV/AIDS and squamous cell carcinoma of the bladder. To complement available treatment programs and break the transmission of disease, sound knowledge and understanding of the biology and ecology of S. haematobium is required. Hybridisation/introgression events and molecular variation among members of the S. haematobium-group might effect important biological and/or disease traits as well as the morbidity of disease and the effectiveness of control programs including mass drug administration. Here we report the first chromosome-contiguous genome for a well-defined laboratory line of this blood fluke. An exploration of this genome using transcriptomic data for all key developmental stages allowed us to refine gene models (including non-coding elements) and annotations, discover 'new' genes and transcription profiles for these stages, likely linked to development and/or pathogenesis. Molecular variation within S. haematobium among some geographical locations in Africa revealed unique genomic 'signatures' that matched species other than S. haematobium, indicating the occurrence of introgression events. The present reference genome (designated Shae.V3) and the findings from this study solidly underpin future functional genomic and molecular investigations of S. haematobium and accelerate systematic, large-scale population genomics investigations, with a focus on improved and sustained control of urogenital schistosomiasis.

Seroprevalence and Associated Risk Factors of Infectious Bronchitis Virus in Chicken in Northwest Ethiopia.

Avian infectious bronchitis virus is a highly contagious disease occurring in respiratory, urogenital, and reproductive tissues of chicken causing considerable losses due to death, egg drop, and reduced production. This preliminary study was conducted to investigate the prevalence of antibodies against infectious bronchitis virus (IBV) and to assess the potential risk factors in chickens of northwest Ethiopia. A cross-sectional study was conducted from November 2020 to June 2021. A total of 768 serum samples from three zones were collected. To investigate the presence of antibodies against IBV, the indirect ELISA serological test was applied. Positivity for anti-IBV antibodies was observed in 23.96% (95% CI: 20.98–27.14) of the samples. The mixed-effect logistic regression analysis of potential risk factors showed that IBV prevalence was significantly higher in young chickens than adults (p < 0.001) and higher in intensive farm type than in extensive type (p < 0.001). Based on the production purposes of the chickens, the odds of seropositivity for IB was significantly higher in layers than in broilers (p < 0.001) and dual purposes (p < 0.001). This study revealed higher seroprevalence in farms which had the “all-in-all-out” rearing method than in farms with different batches in one house with a significant difference (p < 0.001), higher seroprevalence in the poor ventilated type than in good ones (p < 0.001), and higher seroprevalence in the houses that did not remove used litter at all than houses of completely disposed and partially disposed litter (p=0.002). Moreover, disinfection of houses had significant effect on the occurrence of IB. Having personal protective equipment was significantly affecting the occurrence of IB, being higher in the farms that have no wearing clothes and shoe than in those having wearing clothes and shoe (p=0.002). In conclusion, the seroprevalence finding in the present study indicated that the organism is circulating among the population of chickens and high enough to cause significant economic losses Therefore, poultry houses should be cleaned, disinfected, and well ventilated and farm attendants should have separate farm shoe and clothes. Further studies on the virus isolation and molecular characterization of the target gene are needed in the study area.

Determination of In Vitro Antimicrobial Susceptibility for Lefamulin (Pleuromutilin) for Ureaplasma Spp. and Mycoplasma hominis.

Lefamulin is the first of the pleuromutilin class of antimicrobials to be available for therapeutic use in humans. Minimum inhibitory concentrations of lefamulin were determined by microbroth dilution for 90 characterised clinical isolates (25 Ureaplasma parvum, 25 Ureaplasma urealyticum, and 40 Mycoplasma hominis). All Mycoplasma hominis isolates possessed lefamulin MICs of ≤0.25 mg/L after 48 h (MIC50/90 of 0.06/0.12 mg/L), despite an inherent resistance to macrolides; while Ureaplasma isolates had MICs of ≤2 mg/L after 24 h (MIC50/90 of 0.25/1 mg/L), despite inherent resistance to clindamycin. Two U. urealyticum isolates with additional A2058G mutations of 23S rRNA, and one U. parvum isolate with a R66Q67 deletion (all of which had a combined resistance to macrolides and clindamycin) only showed a 2-fold increase in lefamulin MIC (1–2 mg/L) relative to macrolide-susceptible strains. Lefamulin could be an effective alternative antimicrobial for treating Ureaplasma spp. and Mycoplasma hominis infections irrespective of intrinsic or acquired resistance to macrolides, lincosamides, and ketolides. Based on this potent in vitro activity and the known good, rapid, and homogenous tissue penetration of female and male urogenital tissues and glands, further exploration of clinical efficacy of lefamulin for the treatment of Mycoplasma and Ureaplasma urogenital infections is warranted.

S2635 Abnormal Liver Enzymes Attributed to HNF-1b Associated Disease

Introduction: Hepatocyte nuclear factor 1 beta (HNF-1b) associated diseases are genetic conditions caused by an error in the HNF1b gene. This gene regulates early development of organs such as the kidney, genital tract, and pancreas. Common signs/symptoms include kidney findings like cysts, mature onset of diabetes at a young age (MODY), genital abnormalities, as well as abnormal pancreas/liver function. We present a case of abnormal liver enzymes secondary to HNF-1b associated disease. Case Description/Methods: A 22-year-old man was referred for abnormal liver enzymes and suspected NAFLD. Past medical history included surgery for an undescended testicle at age 6 and new onset DM with relative weight loss. Due to DM, weight loss, and a family history of pancreatic cancer he was evaluated by GI and had labs in 1/2020 showing ALK PHOS (254 U/L), AST (98 U/L), ALT (251 U/L), and total bilirubin (1.4 mg/L). MRI/MRCP in 3/2020 was overall unremarkable and upper EUS 6/2020 showed echogenic liver and increased fullness of the pancreas with non-hyperechoic stranding throughout. Detailed serologic evaluation was without specific abnormalities other than ongoing elevated liver enzymes in a mixed pattern. He had a liver biopsy in 1/2021, in which native bile ducts were absent in < 50% of portal tracts, while other portal tracts contained small ducts relative to size of the tract. Portal vein abnormalities were noted including atrophy and fragmentation. No features of NAFLD or chronic liver diseases. He underwent a genetic cholestasis panel showing a heterozygous ∼1.51 Mb deletion that was pathogenic for HNF-1b associated disease. He has been referred to a genetic counselor and maintains follow up with his specialty teams. Discussion: The HNF1B gene is associated with functional and transcriptional regulation in hepatobiliary, pancreatic B-cells, and urogenital tract tissues. Pathogenic deficiencies are related to point mutations or deletions. The combination of MODY 5 diabetes plus renal cysts or urogenital abnormality has led this finding to be called RCAD (renal cysts and diabetes). There have been case reports with classic syndrome plus varying hepatobiliary findings. To our knowledge, this is the first case of HNF-1B associated disease without the classic finding of renal pathology. It is suggested that for young patients with early onset of diabetes with abnormal hepatobiliary and/or pancreatic testing, HNF-1B disease be considered in the differential diagnosis.Figure 1.: Prevention Genetics - Cholestasis Panel Molecular Genetic Report.

Gene Ontology Enrichment Analysis of Renal Agenesis: Improving Prenatal Molecular Diagnosis

Uni- or bilateral renal agenesis (RA) is a commonly occurring major congenital anomaly impacting fetal and neonatal outcomes. Since the etiology is highly heterogeneous, our aim was to provide a logically structured approach by highlighting the genes in which variants have been identified to be associated with RA and to define the pathways involved in this type of abnormal kidney development. We used Phenolyzer to collect a list of all the genes known as causative for RA. Using ClueGO gene enrichment analysis, we classified the relationship between these genes and the biological processes defined by gene ontology. We identified 287 genes and 69 groups of enriched biological processes. About 50% included pathways directly related to the development of urogenital organ tissues. Several ciliary, axis specification, hindgut development, and endocrine pathways were enriched, which may relate to different clinical presentations of RA. Our gene ontology enrichment analysis shows that genes representing distinct biological pathways are significantly enriched. This knowledge will lead to an improved molecular diagnosis in clinical care when applying genome-wide sequencing approaches. The findings will also allow to further study the biological pathways involved in RA and to identify novel candidate genes and pathways.

Angiotensin-converting enzyme 2 gene expression in human male urological tissues: implications for pathogenesis and virus transmission pathways

BackgroundSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection occurs through binding and internalization of the viral spike protein to the angiotensin-converting enzyme 2 (ACE2) receptor on the host cell membrane. Pathological changes are caused by damage and failure of vital organs that express high levels of ACE2, including the lungs, the heart and the kidneys. The aim of this study was to investigate ACE2 gene expression in the human male urogenital tract using a public database.MethodsA search of transcriptomic datasets from a database to investigate ACE2 gene expression in human urogenital tract tissue.ResultsThe gene expression profile demonstrated that ACE2 gene expression was higher in human kidney cortex and testis than human lung tissue. The gene expression profile demonstrated that ACE2 gene expression in the human bladder and prostate was comparable to human lung tissue.ConclusionsMale urogenital tissues are directly susceptible to SARS-CoV-2 infection through the expression of ACE2. Moreover, the SARS-Cov-2/ACE2 interaction may disturb the male genital and reproductive functions.

An advanced method for the immunohistochemical detection of nitric oxide synthase (NOS) in the female genital tract

The expression of nitric oxide synthase (NOS) in male and female urogenital tissues has been investigated by using conventional light microscopical immunoperoxidase staining. We present an improved immunohistochemical method for the specific and simultaneous detection of endothelial and neuronal NOS (eNOS/nNOS) in vaginal tissue. Specific antibodies have been used in combination with the tyramide signal amplification method. We found a subepithelial meshwork of varicose nerve fibers. A subpopulation of fibers presented immunoreactivity specific for nNOS. Epithelial cells also showed cytoplasmatic labeling for nNOS. Arteries presenting signals for eNOS in their endothelial layer were found in close proximity to nNOS-positive nerve fibers.

Methotrexate-induced toxic effects and the ameliorating effects of astaxanthin on genitourinary tissues in a female rat model.

The purpose of the study was to explore the possible deleterious effects of Methotrexate (MTX) treatment on the urogenital tissues and the potential protective effects of Astaxanthin (AXA). Twenty-four female Wistar Albino rats (12months old) were divided into 3 groups as follows: Group I (Control group): rats received a single dose of 0.1ml saline by gavage and intraperitoneal injection. Group II (MTX group): rats received a single dose of 20mg/kg MTX, i.p, on the 2nd day. Group III (MTX + AXA group): rats received 100mg/kg AXA orally for 7days in addition to a single dose of MTX. The levels of total oxidant status (TOS), total antioxidant status (TAS), oxidative stress index (OSI), and histopathological and immunohistochemical markers (Caspase-3, iNOS, CRP, G-CSF) were evaluated in urogenital tissues. In ovarian tissues, a statistically significant increase in TOS levels (p = 0.001) and OSI index (p = 0.028) were observed in Group II compared to Group I. TAS level was significantly higher in Group III compared to Group II and I (p = 0.009 and 0.002, respectively). However, a significant decrease in OSI level was observed in Group III compared to Group II (p = 0.035). In fallopian tube tissues, TAS level was significantly decreased in Group II compared to Group I (p = 0.047). Histopathologically, marked hyperemia was observed in MTX group. AXA treatment ameliorated all the pathological findings. Immunohistochemically, all the studied markers were considerably increased in Group II, however, they were decreased by AXA. These findings revealed that MTX treatment caused oxidative stress, apoptosis, and inflammation in the urogenital tissue. We found that AXA significantly ameliorated the damage caused by MTX in the urogenital tissue. The results of the study have indicated that AXA may be a promising nutritional support substance against the damage caused by chemotherapeutic and cytotoxic agents, such as MTX, to the urogenital tissue.

Evaluation of surgical procedures of mouse urethra by visualization and the formation of fistula

Visualization of the surgically operated tissues is vital to improve surgical model animals including mouse. Urological surgeries for urethra include series of fine manipulations to treat the increasing number of birth defects such as hypospadias. Hence visualization of the urethral status is vital. Inappropriate urethral surgical procedure often leads to the incomplete wound healing and subsequent formation of urethro-cutaneous fistula or urethral stricture. Application of indocyanine green mediated visualization of the urethra was first performed in the current study. Indocyanine green revealed the bladder but not the urethral status in mouse. Antegrade injection of contrast agent into the bladder enabled to detect the urethral status in vivo. The visualization of the leakage of contrast agent from the operated region was shown as the state of urethral fistula in the current hypospadias mouse model and urethral stricture was also revealed. A second trial for contrast agent was performed after the initial operation and a tendency of accelerated urethral stricture was observed. Thus, assessment of post-surgical conditions of urogenital tissues can be improved by the current analyses on the urethral status.