Vaginal Cytology Research Articles

Relationship between thyroid function and sex hormones in female German shepherd dogs.

Several variables influence the serum concentration of thyroid hormones in dogs, including breed, age, drugs, and concurrent diseases. However, data regarding the interaction between thyroid function and the estrous stage of female dogs are limited. Estrous stage may influence thyroid function in German Shepherd dogs. Longitudinal, observational, non-randomized cohort study. The dogs were monitored during the complete estrous cycle, and different stages were determined by vaginal cytology. Two blood samples were collected at the beginning and end of each stage to analyze the following: total thyroxine (TT4), free thyroxine (fT4), total triiodothyronine (TT3), free triiodothyronine (fT3), canine thyrotropin (cTSH), progesterone, 17-β-estradiol, triglycerides, and cholesterol concentrations. Hematological and biochemical evaluations were performed at the beginning and end of the study period. Seventeen German Shepherds were included, of which 7 were bred during the study period. One dog was excluded for estrus interruption and another for suspected hypothyroidism. Serum concentrations of T4, fT4, and fT3 were negatively correlated with age. Total thyroxine demonstrated significant changes in serum concentrations between estrous stages, with higher concentrations in estrus and diestrus. Total thyroxine concentrations were positively correlated with progesterone concentrations and negatively correlated with 17-β-estradiol concentrations. Free thyroxine did not show significant variations but was positively correlated with progesterone concentrations. Canine TSH concentrations were positively correlated with 17-β-estradiol concentrations. No significant differences in thyroid hormones and cTSH concentrations were observed between diestrus during pregnancy and pseudopregnancy. Different stages of estrus can influence the measurement of TT4 in female dogs.

Aged and induced-premature ovarian failure mouse models affect diestrus profile and ovarian features

Sex hormones exert a wide influence on several systems of the human body, especially in women, who undergo intense changes in the trans and postmenopausal periods. Different experimental models are used to mimic these conditions; however, the impact on hormonal profile may be different. This study aimed to analyze and compare vaginal cytology of different post-estropausal mice models, along with their microscopical ovarian features. Forty-six C57BL/6J female mice with the ages of 4, 6 and 18 months at the beginning of the experiment, weighing about 25–28 grams, constituted five groups: NC–(negative control) animals with no treatment, OVX-SHAM—sham ovariectomized, OVX–ovariectomized, VCD–medicated with 160 mg/kg/day of 4-vinylcyclohexene diepoxide via IP for 20 consecutive days, and Aged–senescent mice under physiological estropause. Euthanasia was performed at different periods for the removal of the ovaries, and after diestrus was confirmed by vaginal cytology for 10 consecutive days. For daily vaginal cytology, morphological and histomorphometric microscopic analyzes were performed. Aged mice presented significant increased neutrophils when compared to VCD group, as well as increased cornified epithelial cells when compared to OVX mice, and also increased nucleated epithelial cells when compared to VCD and OVX. NC and OVX-SHAM ovaries presented innumerous follicles at different stages of development, while VCD showed marked follicular atresia, depleted of primordial or developing follicles and a predominance of interstitial cells. The ovaries of aged mice were predominantly constituted by corpus luteum degenerated into corpus albicans, with rare antral follicles. All analyzed models led to different permanent diestrus profiles caused by each model, as indicated by ovarian features. This should be carefully considered when choosing a post-estropausal experimental model, in order to better correlate this challenging phase of female’s life with physiological/pathological conditions.

Vaginal Cytology as a tool for estrus detection improves the success of artificial insemination in water buffalo

Estrus detection in water buffalo is affected by the high incidence of poor manifestation inherent in this animal species. This affected the success rate of Artificial Insemination (AI) and Embryo Transfer (ET) due to the failure of perfect timing in AI and embryo-recipient synchrony in ET. While various ways of estrus detection were reported, the accuracy was interfered with by other symptoms that signify the health condition rather than the reproductive status of the animal. This results in a low conception rate and long calving intervals in AI and ET programs. Vaginal Cytology is a simple clinical tool of estrus detection that can also detect the pathological status of the reproductive tract and the perfect time for AI. While this is greatly used in canines, application in large animals was limited. The need to collect the swab from animals in the field and bring the smeared sample for analysis to the laboratory hindered its use and application. Our study, however, showed that by adopting this tool in estrus detection, increase in conception rate and calf production were achieved. A >60.0% cornification with the presence of >10 mm Graafian follicle is enough indicators of the perfect time for AI resulting in 80-100% conception. The development of a water buffalo vaginal chart helped determine the time of estrus. Strategies to adopt this tool for improved livestock reproduction are warranted.

Abnormal Vaginal Cytology after Total Laparoscopic Hysterectomy in Patients with Cervical Intraepithelial Neoplasia.

To explore the incidence of abnormal vaginal cytology after total laparoscopic hysterectomy for the treatment of cervical intraepithelial neoplasia 3, we retrospectively analyzed the medical records of patients treated at NHO Shikoku Cancer Center (Japan) in 2014-2019. The cases of 99 patients who underwent a laparoscopic (n=36) or open (n=63) hysterectomy and postoperative follow-up were examined. Abnormal vaginal cytology was detected in 13.9% (5/36) of the laparoscopic-surgery (LS) group and 14.3% (9/63) of the open-surgery (OS) group. A vaginal biopsy was performed at the physicians' discretion; one LS patient and six OS patients were diagnosed with vaginal intraepithelial neoplasia. The cumulative incidence of abnormal vaginal cytology at 3 years post-hysterectomy was 21.4% (LS group) and 20.5% (OS group), a nonsignificant difference. A multivariate analysis showed that age > 50 years was the only independent risk factor for abnormal vaginal cytology among the covariates examined including age; body mass index; histories of vaginal delivery, abdominal surgery, and smoking; and surgical approach (hazard ratio 8.11; 95% confidence interval 1.73-37.98; p=0.01). These results suggest that the occurrence of abnormal vaginal cytology after a hysterectomy may not be influenced by the laparoscopic procedure but is associated with older age.

Phytochemical-Based Study of Ethanolic Extract of Saraca asoca in Letrozole-Induced Polycystic Ovarian Syndrome in Female Adult Rats.

Polycystic ovarian syndrome (PCOS) is a complex metabolic and endocrine disorder which affects women of reproductive age. It is a condition in which ovaries produce an excessive amount of androgen (the male sex hormone). Saraca asoca (Roxb.) Willd. is a plant of the Fabaceae family. This plant has been traditionally used as a uterine tonic in leucorrhea and dysmenorrhea due to its various pharmacological activities. In this study, the ethanolic extract of S. asoca (EESA) was evaluated for its potential to be used for the management of PCOS. HPLC analysis revealed the presence of various phytoconstituents: kaempferol, rutin, (-)-epicatechin, salicylic acid, and gallic acid. For PCOS induction, 30 adult female rats were randomly divided into two groups: the control group (n = 5) and the PCOS group (n = 25). Letrozole (1 mg/kg/day) was administered per orally (p.o.) for a period of 7 weeks for the induction of disease. Weekly body weight measurements and daily vaginal cytology examinations were performed for disease confirmation. After disease induction, the PCOS group was further divided into five groups (n = 5), that is, disease control, metformin, and EESA (200, 400, and 600 mg/kg) groups, respectively, and given treatment doses for next 5 weeks. After the treatment period, all animals were weighed and euthanized humanly. Blood samples were collected for hormonal assays, lipid profiles, and liver function tests. For histological assessment of ovarian cysts, ovaries were dissected. Livers were preserved to evaluate EESA's antioxidant properties. Histopathology analysis revealed that EESA reduced body weight and the number of cystic follicles. Furthermore, it also lowered the elevated levels of serum testosterone, luteinizing hormone, insulin, and malonaldehyde in PCOS rats while increasing the levels of follicle-stimulating hormone, estradiol, progesterone, prolactin, and other antioxidant enzymes such as superoxide dismutase, glutathione, and catalase. It can be concluded that EESA exhibited beneficial effects in normalizing the perturbed hormonal profile and improved the ovary status by decreasing the cystic follicle and improving the ovulation status in a dose-dependent manner.

OR01-06 Single Nucleus Multi-omics Analysis Identifies Cellular Trajectories And Dynamic Changes In Gene Expression And Chromatin Accessibility In The Pituitary During The Mouse Estrous Cycle

Abstract Disclosure: F.M. Ruf-Zamojski: None. W. Cheng: None. Z. Zhang: None. M. Zamojski: None. G.R. Smith: None. X. Chen: None. N. Mendelev: None. G. Strupinsky: None. C.A. Alonso: None. L. Ongaro Gambino: None. X. Zhou: None. E. Brule: None. M.S. Amper: None. P. Hanna: None. V.D. Nair: None. C.L. Andoniadou: None. J.L. Turgeon: None. O. Troyanskaya: None. E. Zaslavsky: None. D.J. Bernard: None. S.C. Sealfon: None. Single cell multi-omics datasets provide an unparalleled power to resolve gene regulatory circuits underlying cellular function in complex tissues such as the pituitary gland [1, 2]. To better understand cellular plasticity and dynamics in the mouse pituitary during the estrous cycle in vivo, we performed same-cell single nucleus (sn) multi-omics for gene expression and chromatin accessibility on individual pituitaries collected from mice at 9am on each day of the cycle, as well as at 6pm and 11pm on proestrus and at 2am on estrus to capture surge events. Cycle stage was determined by vaginal cytology and post-mortem measurement of serum LH and FSH levels. In total, 102,069 cells passed rigorous quality control (QC, [2]), with over 5,000 cells analyzed per sample, ∼2,000 genes/cell, ∼15,000 ATAC median high-quality fragments, and Transcription Start Site (TSS) enrichment scores above 9 for all samples. We identified 13 well-separated clusters in the snRNAseq and 10 in the snATACseq datasets representing the pituitary cell types that were followed over time. We integrated the gene expression and chromatin accessibility datasets and analyzed changes in cell type proportions, gene expression, and chromatin accessibility through time. We detected major differential gene expression changes in the gonadotropes and lactotropes, which we further investigated using pseudotime trajectory analyses. Several upstream Fshb loci showed dynamic changes during the estrous cycle. Additionally, we uncovered regulatory components of major pituitary genes over time using linkage data analysis.To our knowledge, this is the first study to present detailed and comprehensive data on gene expression and chromatin structure changes at sn resolution in all pituitary cell types during a dynamic physiological process. Thus, it provides critical new resources to the field of endocrinology.

Successful Management of Follicular Cyst-induced Dermatoses in a Labrador Retriever Dog

Background: Sex hormone dermatoses are uncommon in dog and causes overproduction of one or more of the sex hormones or exogenous administration. Ovarian cyst-induced hyperestrogenemia increased cortisol, hypertestosteronemia, and skin lesions mostly underwent undiagnosed and untreated. This case was documented as clinical pathological changes, diagnosis, and treatment of sex hormone imbalance-induced dermatosis due to an ovarian follicular cyst in a Labrador Retriever bitch. Methods: This clinical investigation was carried out in the month of April’2022 at Veterinary Clinical Complex, Veterinary College and Research Institute, Tirunelveli with a history of prolonged proestrus bleeding, perineal pruritus with reddening, swelling, and enlargement of the vulva. Vaginal exfoliative cytology, hormonal assay, ultrasonography, histopathology of skin, and haemato-biochemistry were performed. Result: Physical examination revealed hyperpigmentation and hyperkeratinization of the skin. Vaginal exfoliative cytology and progesterone assay revealed cytological estrus. Ultrasonography revealed greater than 10 mm-sized anechoic structures in both ovaries along with uterine endometrial hyperplasia. Hormonal analysis revealed hyperestrogenemia and increased cortisol and testosterone. A biopsy of the perineal skin revealed hyperplasia and hyperkeratosis. The case was confirmed as sex hormone imbalance-induced dermatosis due to an ovarian follicular cyst. The animal was treated with two shots of Inj. hCG 500IU in 48 hrs interval followed by OHE after three months.

Bone strength is reduced in a neonatal androgenized rat model

BackgroundWhether polycystic ovary syndrome (PCOS) affects bone health during a woman's lifespan remains controversial. An androgenized rodent model replicated many metabolic and reproductive features of women with PCOS, and we aimed to use it to investigate the impact of androgens on microarchitecture (by micro-CT), bone mechanical strength, bone formation and resorption markers in rats with intact ovaries (SHAM) who underwent oophorectomy. MethodsWistar rats (Rattus norvegicus albinus) were employed for the experiments in this study. The protocol of androgenization consisted of the application of 1.25 mg s.c. testosterone propionate beteween days 2–5 of life, while the controls received the same amount of corn oil s.c. as previously established. Androgenized SHAM rats exhibited chronic anovulation identified by vaginal cytology and a reduction in the proportion of corpus luteum in the ovary in comparison to control SHAM rats. The realization of the ovariectomy or SHAM procedure occurred on Day 100 of life. All groups (n = 8) were followed-up for 180 days to address the study endpoints. ResultsMicro-CT from androgenized female rats (SHAM) showed a divergence between the trabecular and cortical bone profiles. Compared to SHAM controls, these rats had an increase in trabecular bone mass with a diminution in bone resorption C-terminal telopeptide of type 1 collagen (CTX) (p < 0.05), a concomitant decrease in cortical area and thickness in the femur, and a reduction in the strength of the femur on the mechanical test (p < 0.01). ConclusionsOur results suggest that a reduction in the cortical thickness and cortical area observed in PCOS model rats was associated with a reduced strength of the femur, despite increased trabecular formation. Ovariectomy in the androgenized OVX group limited the progression rate of cortical bone loss, resulting in bone resistance and cortical thickness comparable to those observed in the control OVX group.

Comparison of glass and digital slides for cervical cytopathology screening and interpretation.

Cervical cancer is the second most common form of cancer and a leading cause of premature death among women aged 15 to 44 worldwide. In Brazil, there is a high prevalence of infection by the human papillomavirus - HPV. Digital pathology optimizes time and space for reading cervicovaginal cytology slides. We evaluated the feasibility of using whole slide images (WSI) for the routine interpretation of cytology exams. A total of 99 cases of vaginal cytology were selected from a reference laboratory in Northeastern Brazil. Three cytotechnicians participated in the study. Cellular atypia was the one that most presented concordance values. Two observers almost perfectly agreed (k = 0.86 and k = 0.84, respectively) on the negative diagnoses. The performance of the evaluators for NILM (negative for intraepithelial lesion and malignancy) showed high reproducibility and sensitivity in the digital slides, mainly between evaluators A and C. In contrast, the microbiology group showed disagreement between the diagnoses by digital slides and the standard- gold. The concordance between the digital diagnoses and the gold standard for ASCUS was 89%. In the inflammatory category, Spearman's test showed similar results between raters A, B, and C (rs = 0.47, rs = 0.41, and rs = 0.47, respectively). This study reports the diagnostic validation using digital slides in view of the need to optimize the cytology visualization process. Our experience shows good diagnostic agreement between digital and optical microscopy in several analyzed categories, but mainly in relation to cellular atypia and inflammatory processes.

Treatment strategies with vitamin E and C in autologous intraperitoneal ovarian transplantation and its impact on ovarian surface epithelium and follicle reserve

The objective of this study was to assess the impact of Vitamin E (Vit E) and Vitamin C (Vit C) on markers of the oxidant-antioxidant system, ovarian follicle reserves, and the surface epithelium in autologous intraperitoneal ovarian transplantation conducted in rats. The study aimed to investigate how these antioxidants influence various aspects related to transplantation outcomes, including oxidative stress markers, the preservation of follicle reserves, and the condition of the surface epithelium. A total of 20 adult female Wistar Albino rats were included in the study and randomly assigned to four different groups. Group 1, consisting of 5 rats, served as the control group and underwent a surgical procedure where their abdomens were opened and closed without any further intervention. Group 2, also consisting of 5 rats, underwent ovarian transplantation. In Group 3, comprising 5 rats, an intraperitoneal (IP) administration of 20 mg/kg body weight (b.w.) of Vitamin E (Vit E) was given 15 min prior to ovarian transplantation. Lastly, in Group 4, which included 5 rats, an IP administration of 50 mg/kg body weight (b.w.) of Vitamin C (Vit C) was given 15 min before ovarian transplantation. Vaginal cytology was performed in order to monitor the estrus phase in the rats. Biochemically, tissue and serum malondialdehyde (MDA) levels and erythrocyte superoxide dismutase (SOD) levels were measured. Histopathologically, the number of dysplastic changes in the ovarian surface epithelium and primordial, primary, secondary, Graaffian, and atretic follicles were examined. Dysplastic changes in the surface epithelium of Group 2 were found to be significantly higher than in Group 1 and 4 (p < 0.02). In Group 2, the ovarian follicle reserves (primordial, primary, secondary, and Graaffian follicles) were significantly lower than in other groups (p < 0.02). In addition, a significant decrease in SOD levels was found in Group 2 compared to other groups (p < 0.02). The study showed that Vit E and Vit C in autologous intraperitoneal ovarian transplantation preserved the ovarian follicle reserve. Vit C was found to be more effective than Vit E.

Therapeutic effects of human umbilical cord mesenchymal stem cell-derived extracellular vesicles on ovarian functions through the PI3K/Akt cascade in mice with premature ovarian failure.

Premature ovarian failure (POF) mainly refers to ovarian dysfunction in females younger than forty. Mesenchymal stem cells (MSCs) are considered an increasingly promising therapy for POF. This study intended to uncover the therapeutic effects of human umbilical cord MSC-derived extracellular vesicles (hucMSCEVs) on POF. hucMSCs were identified by observing morphology and examining differentiation capabilities. EVs were extracted from hucMSCs and later identified utilizing nanoparticle tracking analysis, transmission electron microscopy, and Western blotting. POF mouse models were established by injecting D-galactose (Dgal). The estrous cycles were assessed through vaginal cytology, and serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), anti-mullerian hormone (AMH), estradiol (E2), and progesterone (P) were measured by ELISA. The human ovarian granulosa cell line KGN was used for in vitro experiments. The uptake of hucMSC-EVs by KGN cells was detected. After D-gal treatment, cell proliferation and apoptosis were assessed via CCK-8 assay and flow cytometry. The PI3K/Akt pathway-related proteins were determined by Western blotting. Our results revealed that POF mice had prolonged estrous cycles, increased FSH and LH levels, and decreased AMH, E2, and P levels. Treatment with hucMSC-EVs partially counteracted the above changes. D-gal treatment reduced proliferation and raised apoptosis in KGN cells, while hucMSC-EV treatment annulled the changes. D-gal-treated cells exhibited downregulated p-PI3K/PI3K and p-Akt/Akt levels, while hucMSC-EVs activated the PI3K/Akt pathway. LY294002 suppressed the roles of hucMSC-EVs in promoting KGN cell proliferation and lowering apoptosis. Collectively, hucMSC-EVs facilitate proliferation and suppress apoptosis of ovarian granulosa cells by activating the PI3K/Akt pathway, thereby alleviating POF.

An immersive simulation strategy to teach canine vaginal cytology: A multi-centre study.

Vaginal cytology (VC) is an essential technique for monitoring the bitch's estrus cycle. Currently, animal-free teaching methodologies have not been investigated for VC. Hence, this study aimed to evaluate an immersive simulation with a VC model and augmented reality tools. Students (n = 219) from four universities were enrolled, having learning stations with models for practising VC that provided immediate feedback on the technique. Augmented reality tools comprised QR codes that endorsed students to short videos of owners' avatars reporting the clinical reproductive story of the simulated animals and slides with QR codes leading to microscopy slide navigation videos. Proestrus, estrus, diestrus, anestrus and vaginitis were identified in the learning stations. The students' perceptions were evaluated through questionnaires assessing satisfaction, motivation, confidence, impact on learning and diagnostic accuracy. Before the immersive simulation, students had no experience with VC, being afraid of doing a VC with a live dog. Almost all the students considered practicing VC as essential and 94% reported that repeating the procedure (>2 times) was the most important parameter for learning. The simulation activity lasted ≈3 h and significantly improved the confidence of students, being less afraid of doing a VC in a live animal. Slide navigation videos improved the diagnostic accuracy of the estrus cycle stage, and students diagnosed the estrus and vaginitis cases more accurately. The immersive simulation strategy allowed repeated practice in a safe, motivated and standardized environment, being appraised by students as an essential strategy for learning VC.

Utility of vaginal vault cytology in the local recurrence of cervical cancer

BackgroundIn Japan, 8000 women were newly diagnosed with cervical cancer in 2018. The healthcare insurance policy in Japan allows physicians to utilize vaginal volt cytology tests and serum biomarker measurement at every visit and imaging analysis at an adequate interval with screening for recurrence after initial treatment. However, the major surveillance guidelines published in the United States and European countries recommend focusing on pelvic examinations and symptom reviews to avoid unnecessary tests. This study aimed to reassess the benefits of standard surveillance methods adopted in this study by retrospective analysis.MethodsFrom January 2009 to December 2015, the medical records of patients with recurrence who were initially diagnosed with International Federation of Gynecology and Obstetrics stage I–III cervical cancer were collected for this study. Clinicopathological data were statistically analyzed to identify significant factors. In the first 2 years, the patients underwent regular surveillance, including pelvic examination, serum tumor marker tests, vaginal vault cytology every 1–3 months, and imaging analysis at 6- to 12-month intervals. In the following 2 years, the patients received a regular check with the same methods every 4 months and an annual imaging analysis. Afterward, the patients had regular screening every 6 to 12 months.ResultsIn the study period, 84 of the 981 patients experienced recurrence, and 88.1% had an asymptomatic recurrence. The disease-free interval was not related to the recurrence site. In univariate analysis, primary treatment, recurrence site, and diagnostic method were significant factors for survival outcomes. In contrast, multivariate analysis indicated that only primary treatment was a significant factor. In patients with local recurrence, multivariate analysis demonstrated that radiation as salvage therapy was an independent predictive factor for overall survival after recurrence.ConclusionsIn this retrospective study, routine imaging analysis and serum biomarker measurement did not contribute to patient prognosis after recurrence. In contrast, vaginal vault cytology can improve survival after recurrence in some patients. Tailored surveillance methods based on individual disease conditions and treatment modalities can improve post-recurrent survival outcomes.

To Study the Vaginal Cytological Changes in Term Pregnant Women and to Determine its role in Predicting the Onset of Labor

Introduction: To study the vaginal cytological changes inpregnant women and to determine its diagnostic accuracy inpredicting the onset of labor.Material and methods: This prospective case study wasconducted in the Department of Obstetrics and Gynecology,SRMS IMS, Bareilly on 100 antenatal patients admitted at term(37–42 weeks). Vaginal cytology was studied in subjects in thepre-labor stage and these subjects were followed in early labor.Results: As the gestation period increases, there is analteration in vaginal cytology under hormonal change. Dueto a decrease in progesterone hormone at term, the numberof intermediate cells decreases and that of superficial cellsincreases under the effect of estrogen hormone. Cluster cellsalso decrease. The sensitivity and specificity of superficialcells was 76.25 and 45%, respectively in predicting labor onsetwithin 3 days.Conclusion: Progesterone

Development and Validation of a Simulation Model for Collection of Canine Vaginal Samples.

Vaginal cytology is a widely used cytological technique mostly taught by observation, either through direct tutoring or videos. To the best of our knowledge, vaginal cytology simulators have never been assessed in veterinary medicine. Twenty-five undergraduate students with no prior experience in canine vaginal sampling were randomly assigned to two groups that either practiced the procedure in a simulator or a live animal. An inverted classroom design was followed. After observing a video tutorial, students practiced with the simulator/live animal for two classes. Three weeks later, they performed a vaginal cytology on a live animal being recorded. The videos were evaluated through an objective structured clinical examination (OSCE) by an observer blinded to the student's groups. The learning outcome was compared through OSCE pass rates and questionnaires. The simulation model was made by 3D printing and soft silicone for the vulvar labia, having pink and blue colored vaseline in the correct and incorrect locations for sampling. The model was economic and accurately replicated the female reproductive tract. It provided immediate feedback to students, by obtaining pink or blue swabs from the correct and incorrect locations, respectively. Students reported that three to five or more attempts were needed to properly learn the procedure, thus justifying the need for a simulator. No differences in the OSCE pass rates were observed between the groups. The simulation model was effective for learning the vaginal cytology procedure, replacing the use of live animals. This low-cost model should be incorporated in the tool-kit of reproduction classes.

Abstract 3210: Monoclonal antibodies targeting PD-1, LAG-3, and TIM-3 do not cause reproductive or endocrine toxicity in a syngeneic model of triple negative breast cancer

Abstract Background: Few targeted treatments exist for triple negative breast cancer (TNBC), an aggressive form of breast cancer that is diagnosed more frequently in reproductive-age women. As a result, most patients receive frontline chemotherapies and often experience infertility and endocrine disruption. Immunotherapies are a promising option for less toxic therapeutic approaches, but little is known about their effects on reproductive and endocrine function. Methods: E0771 cells were injected into the bilateral mammary pads of 8-week-old female C57Bl/6J mice. After tumor development (14 days post-implantation), E0771 mice received 200 ug doses of immunotherapy (anti-PD-1, anti-LAG-3, or anti-TIM-3 antibodies) or IgG isotype control every four days, with treatments ceasing after the third dose. 8 days after the final dose, estrus cycling was monitored via vaginal cytology. 11-15 days after the final dose, mice were euthanized during proestrus stage, and serum and ovaries were collected. Serum levels of anti-Mullerian hormone (AMH), luteinizing hormone (LH), and follicle stimulating hormone (FSH) were quantified via ELISA. Ovaries were fixed in formalin and embedded in paraffin, then sectioned serially. Follicle staging and abundance was quantified via hematoxylin and eosin staining. Follicle health and apoptosis was evaluated via TUNEL staining. Results: There were no significant differences between any of the treatment and control groups in the percentage of time spent in each stage of the estrus cycle, indicating that treatment with the selected immunotherapies does not cause perturbations in reproductive cyclicity. Serum hormone analysis revealed no significant differences in AMH, LH, or FSH levels between treatment and control groups, indicating that these immunotherapies do not cause endocrine toxicity. There were no significant differences in follicle abundance between any of the treatment and control groups, suggesting that the immunotherapies are not toxic to the ovarian reserve or developing follicles. TUNEL staining revealed no significant differences in granulosa cell or oocyte apoptosis between treatment groups. Conclusion: Treatment with anti-PD-1, anti-LAG-3, or anti-TIM-3 monoclonal antibodies does not cause reproductive or endocrine toxicity in a tumor-bearing mouse model. These results demonstrate that immunotherapy may be a promising component of fertility-sparing therapeutic regimens for patients who wish to maintain ovarian and endocrine function after cancer treatment. Citation Format: Payton De La Cruz, Jennifer Ribeiro, Kathryn Grive. Monoclonal antibodies targeting PD-1, LAG-3, and TIM-3 do not cause reproductive or endocrine toxicity in a syngeneic model of triple negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3210.

Dose Dependent Effects of Aqueous Extract of Garcinia cambogia Desr. Against Letrozole Induced Polycystic Ovarian Syndrome in Female Adult Rats With Possible Mechanisms Exploration.

Polycystic ovarian syndrome (PCOS) is an endocrine metabolic disorder of women. This study aimed to explore the potential of aqueous extract of Garcinia cambogia Desr. (AEGC) in PCOS. The HPLC was used to determine the phytoconstituents present in Garcinia cambogia. Thirty adult female albino rats were divided into 6 groups: Normal control (NC) disease Control (PCOS; letrozole 1mg/kg), plant extract (AEGC 100, 300, 500mg/kg) and standard (metformin; 20mg/kg). Disease was confirmed by vaginal smear cytology. After 10 weeks, animals were euthanized, ovaries dissected for histopathology, blood collected for hormonal and biochemical analysis. HPLC analysis showed the presence of phenolic contents; chlorogenic acid, gallic acid, coumaric acid while flavonoid contents were quercetin, kaempferol, and rutin. After treatment, there was dose dependent reduction of weight, ovarian cysts, improvement of follicle growth. DPPH radical scavenging percentage was 67.89%. Hormonal analysis showed a significant improvement (P < .05) in follicle stimulating hormone (FSH), estrogen, and progesterone while a reduction in testosterone, luteinizing hormone (LH) and insulin level. Antioxidant enzymatic markers were significantly (P < .05) increased. Lipid profile and LFTs were also improved. The study validated the potential of Garcinia cambogia in the management of PCOS.

The Track-by-Day Method for Monitoring the Rodent Estrous Cycle.

The exclusion of female subjects from preclinical neuroscience research has traditionally been justified in part by concerns about potential effects of cycling ovarian hormones on brain function. There is evidence that some behavioral and neurobiological measures do change over the estrous cycle and, as the use of female subjects becomes increasingly routine, there is a greater demand for accessible cycle-tracking methods. Conventional estrous cycle staging requires expert training in the qualitative interpretation of vaginal cytology smears, which serves as a barrier for novice researchers. In addition, definitions and reporting practices are not standardized across laboratories, which makes it difficult to compare results across studies and likely contributes to a false perception of the cycle as ephemeral and inconsistent. Here, we describe a streamlined method for monitoring the estrous cycle in rats, which we term Track-by-Day. It is simple to implement and inherently produces consistent reporting. Our protocol should serve to demystify and facilitate adoption of cycle tracking for those new to the practice. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Collection and staining of vaginal smears Basic Protocol 2: Track-by-Day classification of vaginal smears Support Protocol: Preparation of gelatin-subbed slides.

Importance of Induction Endometritis and Vaginal Cytology in Rat Model: A Narrative Review Article

Introduction: Endometritis is a female reproductive disease characterized by the inflammation of the endometrial glandular and stromal tissues. It is usually occurred due to uterine contamination with infectious causes. Animal models of clinical endometritis are very important to prevent the incidence and treatment of endometritis. Vaginal cytology method is considered as a golden standard for determining the estrous cycle stage in female rats. The aim of this study was to induce endometritis in a rat model and determine the stages of the estrous cycle of this animal using vaginal cytology method in order to successfully induce this condition in this animal model.&#x0D; Conclusion: Inflammation changes the production of endometrial cytokines, which may result in menstrual disorders and decreased endometrial reception by impaired endometrial tissue dysfunction. Increased expression of specific cytokines genes such as IL-6, IL-8 and TNF-α can be used to detect the severity and degree of endometritis. Uterine infections are known as a result of damage to the defense mechanisms of the uterine tissue, which are caused by physiological changes in the postpartum period. Another change that occurs in the postpartum period is the increase in the production of reactive oxygen species. The imbalance between the production of reactive oxygen species and its clearance leads to the creation of oxidative stress conditions. On the other hand, there is a significant relationship between inflammatory diseases such as endometritis and oxidative stress. On other hand, by accurately determining the estrous cycle of rats by using vaginal cytology, endometritis can be well induced in this species.

Uterine issues in infertile queens: Nine cases

This prospective case series investigated potential uterine causes of infertility in queens. Purebred queens with infertility (failure to conceive, embryonic death, or failure to maintain pregnancy and produce viable kittens), but no other reproductive disorders were examined approximately 1–8 weeks before mating (Visit 1), 21 days after mating (Visit 2), and 45 days after mating (Visit 3) if pregnant at Visit 2. Investigations included vaginal cytology and bacteriology, urine bacteriology, and ultrasonography. At Visit 2 or 3, uterine biopsy or ovariohysterectomy was performed for histology. Of nine eligible queens, seven were non-pregnant by ultrasound at Visit 2 and two had lost pregnancies by Visit 3. Ovulation was confirmed by serum progesterone concentration in all queens. Ultrasonic appearance of the ovaries and uterus was compatible with a healthy status except for one queen with signs of cystic endometrial hyperplasia (CEH) and pyometra, a follicular cyst in another, and fetal resorptions in two queens. Six cats had histologic lesions of endometrial hyperplasia, including CEH (n=1). Only one cat had no histologic uterine lesions. Bacteria were cultured from vaginal samples in seven queens at Visit 1, (two were non-evaluable), and in five of seven queens sampled at Visit 2. Uterine cultures were negative except for the cat with pyometra. All urine cultures were negative. In summary, the most frequent pathology observed in these infertile queens was histologic endometrial hyperplasia, which can potentially inhibit embryo implantation and healthy placental development. This suggests that uterine disease might contribute substantially to infertility in purebred queens.