Vaginal Cytology Research Articles

Comparing Urtica dioica to Placebo on Vaginal Cytology and pH in Postmenopausal Women: A Randomized, Controlled Trial

IntroductionDecreased estrogen in the period of menopause causes thinning of the vaginal lining, a rise in vaginal acidity, and a drop in vaginal maturation levels. Menopausal women are seeking treatment for bothersome vaginal symptoms, opting for herbal treatments instead of hormone therapy due to their side effects or contraindications. MethodsIn 2021-2022, a triple-blind randomized placebo-controlled clinical trial was carried out on 84 postmenopausal women aged 45-60 from Golestan who had the eligibility criteria and sought services from comprehensive health centers. The qualified females were given 5% vaginal cream containing Urtica dioica extract and a placebo for a duration of 8 weeks. Analysis of data was performed with SPSS. Vaginal PH and maturation value were measured before, as well as four and eight weeks following the intervention. ResultsBefore the intervention, there was no significant difference in the mean (SD) scores of the vaginal pH and VMV between the two groups. During the fourth and eighth weeks post-intervention in the Urtica dioica group, the mean (SD) vaginal pH scores were 4.8 ± 0.7 and 4.3 ± 1.0, while the mean (SD) VMV scores were 50.8 ± 8.8 and 51.3 ± 8.9, respectively. There was a significant difference between the intervention group and the placebo group (P<0.001). ConclusionsAfter using Urtica dioica vaginal cream for eight weeks, postmenopausal women displayed higher vaginal maturation value and lower vaginal pH.

Determination of vaginal cytology and MMP-9 expression during early pregnancy in mice

Determination of vaginal cytology and MMP-9 expression during early pregnancy in mice

The Influence of the Estrous Cycle on Neuropeptide S Receptor-Mediated Behaviors.

The Neuropeptide S receptor (NPSR) has been identified as a potential therapeutic target for anxiety and post-traumatic stress disorder. Central administration of Neuropeptide S (NPS) in male mice produces anxiolytic-like effects, hyperlocomotion, and memory enhancement. Currently, the literature is limited in the number of studies investigating the effects of NPS in female test subjects despite females facing a higher prevalence of anxiety-related pathology, as well as greater risk for adverse effects while taking psychoactive drugs. Moreover, no previous studies have considered the influence of estrous cycle on the effects of NPS. The current study investigates whether NPS-mediated behavioral phenotypes seen in males translate to females, and whether they are affected by estrous cycle stage. Female C57BL/6NCr mice were intracerebroventricularly (ICV) cannulated and underwent behavioral paradigms to test locomotion, anxiety, and memory. Estrous cycle stage was determined through examination of vaginal cytology. Our results provide evidence that NPS-mediated behaviors are influenced by the estrous cycle. Administration of NPS decreased anxiety-like behaviors more robustly when the female mice were in high estrogen stages of the estrous cycle. Therefore, the desired anxiolytic-like effects of targeting the NPSR are intact in female mice. However, these effects may to be influenced by the stage of the estrous cycle. The NPSR remains a strong potential drug target for new anxiolytic compounds and based on our initial observations further studies exploring the interaction of estrous cycle and the NPS-system are warranted. Significance Statement The neuropeptide S (NPS) receptor has been identified as a potential target for treating anxiety, a condition that is most prevalent in females. Therefore, the potential interaction of estrous cycle with the NPS-system described in the current study is an important first step in understanding the function of the NPS-system in females.

Vaginal cytology during the estrous cycle and early pregnancy of ewes

Changes in vaginal epithelial cells and blood progesterone and estrogen concentrations were compared between estrous cycle and early pregnancy in multiparous Sanjabee ewes. Twenty non-pregnant ewes were synchronized with intravaginal insertion of a controlled internal drug release (CIDR) device which was in place for 7 days, intramuscular administration of a GnRH analog (Alarelin acetate; 12.5 µg) at the time of CIDR insertion (Day -7), a PGF2α analog (D-cloprostenol sodium; 125 µg) and 500 I.U. of human chorionic gonadotropin (hCG) at the time of CIDR removal (Day 0). At Day 0, all ewes were introduced to four fertile rams and observed for estrous signs. Ewes exhibiting estrous signs were divided into two groups: MATED (confirmed pregnant with transrectal ultrasound scanning at Day 35) and NOT-MATED (confirmed non-pregnant with transrectal ultrasound scanning at Day 35). Vaginal smears and blood samples were taken daily and every other day, respectively, from the beginning of estrus for 20 days for cytology and hormone assay. Based on the results, during days 0 and 1 of the estrous cycle, only the percentages of intermediate cells differed between the groups (P &lt; 0.05). During days 2 to 4, there was no difference in the cell populations between the groups (P ≥ 0.05). During days 5 to 16, the percentages of all cell types except the parabasal cells differed between the groups (P &lt; 0.05). During days 17 to 20, the percentages of all types of vaginal epithelial cells differed between the groups (P &lt; 0.05). Progesterone concentrations increased gradually from days 0 to 14 of the estrous cycle in both groups, however, they decreased significantly afterward in the NOT-MATED group. Estrogen concentrations changes showed an opposite pattern to that of progesterone in the study groups. Collectively, vaginal cytology can be used as a useful tool in assessing hormonal and physiological characteristics of the female reproductive system and thus provides a more accurate understanding of the physiology of estrous cycle and early pregnancy in ewes, which can be used to improve reproductive management.

Clinical trial comparing the use of Orcellex® Brush versus Cervex-Brush® on vaginal vault smear cytology adequacy rate in patients treated with radiotherapy for cervical cancer.

This study aims to evaluate and compare the Cervex-Brush® and Orcellex® Brush as sampling devices for vaginal vault smear cytology in cervical cancer patients treated primarily with radiotherapy. A randomized crossover trial was conducted at a gynecological oncology center in Hong Kong to compare the Cervex-Brush® and Orcellex® Brush in terms of their vault smear adequacy rate in cervical cancer patients who underwent radiotherapy. One hundred sixty cervical cancer patients treated with primary radiotherapy and undergoing follow-up surveillance by vaginal vault cytology were recruited. The smear adequacy rate was 90.6% for Cervex-Brush® and 91.9% for Orcellex® Brush. The rates of low cellularity for both brushes were similar (76.8% for Cervex-Brush® vs. 76.1% for Orcellex® Brush). The detection rates of abnormal smears were also not significantly different (2.8% for Cervex-Brush® vs. 4.2% for Orcellex® Brush). The 2 brushes were also not significantly different in terms of pain score and degree of bleeding. It was further observed that the second smear collection was more painful and patients who were on hormonal replacement therapy demonstrated less bleeding. There was no difference between the Orcellex® brush and the Cervex-Brush® in terms of smear adequacy rate, rate of high cellularity and the detection of abnormal smears. There was also no significant difference between the 2 brushes in terms of pain and the degree of bleeding. Therefore, the Orcellex® Brush can be considered a suitable alternative sampling device for vault smear collection in patients who have undergone radiotherapy for cervical cancer. ClinicalTrials.gov Identifier: NCT04461574.

7869 Conditional deletion of androgen receptors from kisspeptin neurons only partially rescues the reproductive phenotype of a prenatal androgen-induced mouse model of polycystic ovary syndrome

Abstract Disclosure: A.G. Novak: None. A.V. Petriv: None. D.B. Lohr: None. K.M. Dillon: None. N.T. Neifert: None. A.M. Moore: None. Polycystic ovary syndrome (PCOS) is a prevalent cause of infertility in people with ovaries of reproductive age. Although the etiology of PCOS remains unclear, research in the clinic and with animal models suggests that hyperandrogenism (the presence of elevated circulating androgens) plays a significant role in the development of reproductive dysfunction, including irregular menstrual/estrous cycles and altered ovarian morphology. Although the site at which androgens disrupt the reproductive axis is unclear, neurons expressing kisspeptin are crucial regulators of the hypothalamic-pituitary-gonadal axis and express androgen receptors. Therefore, we hypothesize that androgen signaling at kisspeptin cells is a critical mediator of reproductive dysfunction in PCOS. To test this, we utilized Cre-lox technology to conditionally delete androgen receptors from kisspeptin cells (KARKO) in a commonly used mouse model of PCOS induced by using prenatal dihydrotestosterone treatment (prenatal androgen-treated (PNA) mice). To assess estrous cyclicity, vaginal cytology was examined daily for three weeks beginning at 50 days of age. Prenatal vehicle-treated (PNV) wild-type (WT) and PNV KARKO mice (n=6/group) exhibited regular estrous cycles with no significant differences in the percentage of time spent in proestrus, estrus, or metestrus/diestrus, indicating androgen receptors are not required to maintain fertility. As expected, PNA WT mice (n=6) rarely entered proestrus, spent significantly less time in estrus (p&amp;lt;0.05), and spent significantly more time in metestrus and diestrus (p&amp;lt;0.001) compared to the PNV groups (n=6/group, two-way ANOVA), replicating the previously reported acyclic phenotype of PNA mice. No significant differences in the percentage of time spent in cycle stages were detected between PNA WT and PNA KARKO (n=5) mice in this analysis, suggesting androgen signaling at kisspeptin cells is not required to induce acyclicity in PNA mice. Despite this acyclic phenotype, preliminary data using ovarian sections (5μm thickness) stained with hematoxylin and eosin revealed that the percentage of ovarian area composed of corpora lutea was significantly elevated in PNA KARKO mice (n=3) compared to PNA WT mice (n=4) (p&amp;lt;0.05, one-way ANOVA). Further, in this analysis, the area composed of corpora lutea in PNA KARKO mice was not significantly different from PNV WT and KARKO controls (n=6/group), suggesting PNA KARKO mice exhibit a comparable ovulation rate to fertile controls. Together, these data show that androgen signaling at kisspeptin cells partially contributes to the development of anovulation in PNA mice. However, additional mechanisms likely contribute to developing the full PCOS-like reproductive phenotype. Presentation: 6/1/2024

A Novel Intravaginal Contraceptive Plug for Cats: A Preliminary Biocompatibility Assessment on Haematology and Vaginal Swab.

This preliminary study evaluated the biocompatibility of a novel degradable intravaginal plug contraceptive composed of PEG 4000 and chitosan in cats using haematological profiling and vaginal cytology. Five healthy, non-pregnant female cats were fully anaesthetised and fitted with an intravaginal plug (10 × 0.3 mm) using an applicator, following oestrogen administration 3 h prior. Blood samples were collected from the cephalic vein on days 0 (pre-insertion) and 3 and 7 (post-insertion). Vaginal cytology examinations were conducted on day 0 (pre- and post-oestrogen injection) and days 1, 3 and 7 post-insertion. Haematological parameters, including red blood cell count, haemoglobin levels, haematocrit values, total white blood cell count and differentiation, showed no significant changes after contraceptive insertion (p > 0.05). Vaginal cytology indicated an acute inflammatory response in one out of five subjects on day three post-insertion. The distribution of vaginal epithelial cells (parabasal, intermediate and superficial) remained unaffected by contraception. Oestrogen injection resulted in the dominance of superficial cells up to day 7 of observation (p < 0.05). Overall, PEG 4000 and chitosan-based intravaginal plug contraceptives demonstrated sufficient biocompatibility, indicating their potential as viable contraceptive options for feline use.

Duration of efficacy and effect of implant location in adult queens treated with a 9.4 mg deslorelin subcutaneous implant

Deslorelin is a GnRH-agonist used off-label for contraception in female cats. Little is known about duration and safety of the 9.4 mg subcutaneous implant in the queen as well as its efficacy when placed periumbilically. Fourteen female cats were administered the 9.4 mg deslorelin implant (during interestrus or anestrus) either in interscapular (N=8) or periumbilical (N=6) sites, following general and reproductive examination, vaginal cytology, hematology, biochemistry and progesterone assay to ensure health status. All above procedures (except for progesterone assay) were repeated weekly during the first month, then every 2 months until 6 months and then every 6 months until treated cats regained full reproductive function. No side effects were observed in any treated queen. Post implantation estrus occurred in 40% of the subjects. A significant increase in body weight was observed during treatment (12/14 queens gained weight), particularly at the end of the study. At the end of the study some queens mated, conceived and kittened, proving reversibility of the treatment. The average duration of action of the 9.4 mg deslorelin implant was 790 +/- 155 days (range 525-1140 days) with no significant difference in duration or efficacy depending on implantation sites. The 9.4 mg deslorelin implant causes pharmacological sterilization for about 2 years in female cats, is fully reversible and caused no clinically relevant side effects when administered at both interscapular and periumbilical sites.

Measuring and exploring variations of serum anti-Müllerian hormone (AMH) in female fishing cat (Prionailurus viverrinus) and leopard cat (Prionailurus bengalensis)

Serum concentration of anti-Müllerian hormone (AMH) is a way to estimate the ovarian reserve in many species including domestic cats. It could therefore become an important indicator to manage breeding of wild cat species in captivity. This study focused on 1) validating the AMH Gen II ELISA test, and 2) exploring the relationship between serum AMH with age and estrus stages in fishing cats (Prionailurus viverrinus) and leopard cats (Prionailurus bengalensis). Blood samples were collected from 11 female fishing cats (5.7 ± 3.2 years old) and 8 female leopard cats (8.1 ± 1.8 years old) subjected to routine annual health check in zoo facilities. The stage of the estrus cycle was estimated from animal history, abdominal sonography and vaginal cytology. Serum AMH was measured by AMH Gen II ELISA. Intra-assay CV and inter-assay CV were 4.11% and 5.52%, respectively. Validation of the AMH Gen II ELISA by the linearity of sample dilution in fishing cats and leopard cats was confirmed. In fishing cats, r2 equalled 0.998 and percentage recovery was 83 to 101%. In leopard cats, r2 equalled 0.999 and percentage recovery was 96 to 104%. Serum AMH concentration ranged from 0.17 to 30.53ng/mL (averaged 14.88 ± 10.03ng/mL) in fishing cats and 1.25 to 13.52ng/mL (averaged 6.25 ± 4.69ng/mL) in leopard cats. There was no significant relationship between serum AMH concentration and age in both fishing cats (r = -0.556, P = 0.075) and leopard cats (r = 0.010, P = 0.982). Likewise, there was no difference in AMH concentrations between estrus stage and other stages in both fishing cats (P = 0.069) and leopard cats (P = 0.599). In conclusion, because the limited sample size and the lack of precise information on the estrus phase, the AMH Gen II ELISA shows promising potential for detecting concentrations that may be correlated with age, estrus cycle, and ovarian reserve in future studies.

Effect of Cucurbita pepo (Pumpkin) seed extracts on the cyclicity and reproductive hormones of female Wistar rats

Introduction: Despite the traditional use of Cucurbita pepo seed in pregnancy, its effects on female reproduction remain scarce. This study evaluated the impacts of n-hexane, dichloromethane (DCM), and aqueous ethanol extracts of C. pepo seed on the cyclicity and reproductive hormones of female Wistar rats. Methods: Ten groups of four rats received seed extracts or tween 80 orally for 21 days: A (control)= 0.5 mL tween 80 (vehicle); B, C, &amp; D= 142.86, 285.71, and 428.57 mg/kg nHE; E, F, &amp; G= 142.86, 285.71, 428.57 mg/kg of DCM; and H, I, &amp; J= 142.86, 285.71, 428.57 mg/kg of aqueous ethanol extracts, respectively. Vaginal cytology monitored the estrous cycle daily, and blood samples were obtained for follicle-stimulating hormone (FSH), luteinizing hormone (LH), estrogen, and progesterone at various oestrus cycle phases. Results: Compared to the control, the estrous cycle phases did not change significantly (P&gt;0.05). FSH levels significantly increased (P&lt;0.05) with DCM and aqueous ethanol extracts of C. pepo seed during proestrus and estrus phases compared to the control. A significant (P&lt;0.05) increase in LH was observed with n-hexane, DCM, and aqueous ethanol extracts during all estrous cycle phases compared to the control. All extracts significantly increased estrogen levels (P&lt;0.05) during all phases. DCM and aqueous ethanol extracts reduced substantially the estrus-phase progesterone. Conclusion: Cucurbita pepo may stimulate the hypothalamic-pituitary-gonadal axis in female reproduction. Further studies should be conducted using various phytoestrogen compounds to gain useful knowledge about the effectiveness, safety, and long-term effects of C. pepo seed extracts in regulating hormonal balance.

Influencing and Regulating the Estrous Cycle in Outbred CD1 Laboratory Mice (Mus musculus)

AbstractThe Whitten effect is a widely used tool for manipulating the mouse estrous cycle and generating reproductively active females within the laboratory setting. Typically, peak numbers of sexually receptive mice occur following exposure to male pheromones, resulting in a higher number of successful copulations on the third day after exposure. Although this method has improved efficiencies, the percentage of females mated and subsequently deemed to be pregnant/pseudopregnant remains relatively low, around 50%. In experiment 1, we aimed to 1) further understand cyclicity; 2) determine whether the initial cycle stage plays an importance on day 3 receptivity; and 3) identify any repetitive patterns/cycle stabilization. Mice (n = 27) were assigned to group cages according to cycle stage (proestrus, estrus, metestrus, diestrus). Experiment 2 was developed to determine an optimum treatment to promote receptivity by exposure to various pheromone stimuli. Mice (n = 45) were randomly assigned to 5 treatment groups (PBS-treated sham soiled bedding, male soiled bedding, live male, pregnant females, and lactating females). In both experiments, daily vaginal cytology was performed for 21 days to determine the cycle stage. Results from experiment 1 indicate that the initial cycle stage did not contribute to day 3 receptivity, although synchronization within several groups/cages was noted, and that the greatest numbers of estrous animals were obtained on days 6 and 7. Experiment 2 revealed that exposure to live males and lactating females both significantly improved receptivity compared with the PBS, male soiled bedding, and pregnant female groups. These results indicate that current strategies used for routine synchronization could be further improved through alternative housing regimens without compromising animal welfare.

Staging of Estrous Cycle in Rats using Vaginal Cytology and Visual Assessment

The determination of different phases of estrous cycle in lab animals is important for reproductive interventions and other experiments related to animal reproduction where rats are used as experimental models. A total 21 Wistar albino (which had completed their pubertal stage ( greater than 4 weeks) weighing approximately 100- 200 g were used in this experiment. Visual assessment was done to observe vaginal opening during different phases of the estrous cycle. This was followed by collection of vaginal discharge samples by flushing it with 0.1 ml of distilled water using micropipette. Vaginal cytology was performed in 21 rats thrice daily, along with visual assessment. The average length of estrous cycle in rats was recorded to be 4-5 days with average duration of proestrus, estrus, metestrus and diestrus of 24, 24-36, 12 and 48 hours, respectively. The average length of estrous cycle in rats was 4-5 days with recorded average duration of proestrus, estrus, metestrus and diestrus of 24, 24-36, 12 and 48 hours, respectively. Vaginal cytology can be considered as a better option compared to visual assessment for assessment of reproductive status in rats.

Augmenting interpretation of vaginoscopy observations in cycling bitches with deep learning model

Successful identification of estrum or other stages in a cycling bitch often requires a combination of methods, including assessment of its behavior, exfoliative vaginal cytology, vaginoscopy, and hormonal assays. Vaginoscopy is a handy and inexpensive tool for the assessment of the breeding period. The present study introduces an innovative method for identifying the stages in the estrous cycle of female canines. With a dataset of 210 vaginoscopic images covering four reproductive stages, this approach extracts deep features using the inception v3 and Residual Networks (ResNet) 152 models. Binary gray wolf optimization (BGWO) is applied for feature optimization, and classification is performed with the extreme gradient boosting (XGBoost) algorithm. Both models are compared with the support vector machine (SVM) with the Gaussian and linear kernel, k-nearest neighbor (KNN), and convolutional neural network (CNN), based on performance metrics such as accuracy, specificity, F1 score, sensitivity, precision, matthew correlation coefficient (MCC), and runtime. The outcomes demonstrate the superiority of the deep model of ResNet 152 with XGBoost classifier, achieving an average model accuracy of 90.37%. The method gave a specific accuracy of 90.91%, 96.38%, 88.37%, and 88.24% in predicting the proestrus, estrus, diestrus, and anestrus stages, respectively. When performing deep feature analysis using inception v3 with the same classifiers, the model achieved an accuracy of 89.41%, which is comparable to the results obtained with the ResNet model. The proposed model offers a reliable system for identifying the optimal mating period, providing breeders and veterinarians with an efficient tool to enhance the success of their breeding programs.

Vaginal cytomorphological profile in correlating suicidal deaths in medicolegal autopsy cases

Objectives: Vaginal cytology is a sensitive method for assessing women’s hormonal status, showing variations in menstrual cycle phases. Studies suggest a higher rate of female suicides during the luteal phase. This study aimed to link vaginal cytomorphological profiles with the manner of death and age of the deceased. Materials and Methods: Vaginal smears from 64 autopsy cases were microscopically examined for maturation index (MI) and value, reflecting hormonal profiles. Statistical Analysis: ANOVA between the mean of maturation value with manner of death was found to be significant. Results: Among the 48 suicides, 54.2% showed predominantly intermediate cells in the luteal phase, indicating a mid-zone shift in the MI. The remaining 16 non-suicidal cases showed fewer intermediate cells. Suicides shared a similar cytological profile and MI, especially in the reproductive age group. Conclusions: While vaginal smears are typically used in sexual assault cases to detect sperm, they also offer insights into women’s hormonal profiles. Understanding these profiles could provide clues about psychological status and its relation to the manner of death beyond sexual assault.

The Estrous Cycle Coordinates the Circadian Rhythm of Eating Behavior in Mice.

The estrous cycle regulates rhythms of locomotor activity, body temperature, and circadian gene expression. In female mice, activity increases on the night of proestrus, when elevated estrogens cause ovulation. Exogenous estradiol regulates eating behavior rhythms in female mice fed a high-fat diet, but it is unknown whether endogenous estrogens regulate eating rhythms. In this study, we investigated whether diurnal and circadian eating behavior rhythms change systematically across the estrous cycle. We first studied diurnal eating behavior rhythms in female C57BL/6J mice in 12L:12D. Estrous cycle stages were determined by vaginal cytology while eating behavior and wheel revolutions were continuously measured. The mice had regular 4- to 5-day estrous cycles. Consistent with prior studies, the greatest number of wheel revolutions occurred on the night of proestrus into estrus when systemic levels of estrogens peak. The amplitude, or robustness, of the eating behavior rhythm also fluctuated with 4- to 5-day cycles and peaked primarily during proestrus or estrus. The phases of eating behavior rhythms fluctuated, but not at 4- or 5-day intervals, and phases did not correlate with estrous cycle stages. After ovariectomy, the eating behavior rhythm amplitude fluctuated at irregular intervals. In constant darkness, the amplitude of the circadian eating behavior rhythm peaked every 4 or 5 days and coincided with the circadian day that had the greatest number of wheel revolutions, a marker of proestrus. These data suggest that fluctuations of ovarian hormones across the estrous cycle temporally organize the robustness of circadian eating behavior rhythms so that it peaks during ovulation and sexual receptivity.

Vaginal Intraepithelial Neoplasia (VaIN) after Hysterectomy Is Strongly Associated with Persistent HR-HPV Infection.

This retrospective study was conducted on 170 women who underwent a laparoscopic hysterectomy due to high-grade cervical intraepithelial neoplasia (CIN3) or benign gynecological disease. The follow-up strategy included performing a cotest and colposcopy with biopsy if necessary. The median time between primary treatment and a diagnosis of high-grade VaIN was 18 months. High-grade or severe VaIN was found in eight patients after hysterectomy (4.7%). All cases of high-grade VaIN occurred in women with persistent HPV infection. The most frequent genotype was 16. Women hysterectomized due to CIN3 showed an eight-fold greater risk than women hysterectomized due to benign disease of developing high-grade VaIN. The risk of VaIN is low in women hysterectomized due to benign disease. The risk of developing VaIN is greater in women with viral persistence. All these elements suggest that it is a history of HPV-related disease of the lower genital tract and viral persistence, rather than hysterectomy itself, that should be considered risk factors for the development of high-grade VaIN. After hysterectomy, patients with a history of CIN should undergo annual screening with vaginal dome cytology and HPV testing.

Machine learning for determining the stage of the estrous cycle in bitches

Reproductive biotechnologies, such as artificial insemination, are important tools in the reproduction of female dogs. Accurate determination of the specific stage of the estrous cycle is crucial for the successful application of these technologies.Vaginal cytology serves as a cost-effective and rapid diagnostic solution. However, itrelies on the analyzer’s expertise, making itsubject to human errors. Additionally, it may involve a prolonged duration between sample collection and result analysis.To minimize these limitations and streamline the diagnostic process,this studydeveloped software to automate the identification of the main phases of the estrous cycle that are important for artificial insemination. Eighteen vaginal cytology images were used, withsix images representing each of the phases studied (proestrus, estrus, and diestrus).Imageswere analyzed using the open source CellProfiller software, with subsequentclassification of the images using theTanagra software. Sensitivity and specificity valueswere determined for the proestrus, estrus, and diestrus phases, yielding results of 0.99, 0.86, 0.95, and 1, 0.95, 0.82, respectively.These findingsdemonstrate the model’scapacityfor correctly identifyingdifferent phases of the estrous cycle. The proposed model proved effective for the study’sobjective, and the authors suggest that it may be applicable to other economically important species.

Biological Sex Influences Daily Locomotor Rhythms in Mice Held Under Different Housing Conditions.

Daily rhythms are programmed by a central circadian clock that is modulated by photoperiod. Here, we recorded locomotor activity rhythms in C57Bl/6 or mPer2Luc mice of both sexes held under different housing conditions. First, we confirm that the structure of locomotor activity rhythms differs between male and female mice in both genetic backgrounds. Male mice exhibit a nightly "siesta," whereas female mice fluctuate between nights with and without a nightly siesta, which corresponds with changes in locomotor activity levels, circadian period, and vaginal cytology. The nightly siesta is modulated by the presence of a running wheel in both sexes but is not required for the infradian patterning of locomotor rhythms in females. Finally, photoperiodic changes in locomotor rhythms differed by sex, and females displayed phase-jumping responses earlier than males under a parametric photoentrainment assay simulating increasing day length. Collectively, these results highlight that sex and sex hormones influence daily locomotor rhythms under a variety of different environmental conditions.

Proliferation and apoptosis dynamics of the normal canine mammary gland during the oestrous cycle evaluated by Ki-67 and Caspase-3 indexes.

Understanding the normal physiology of the canine mammary gland (CMG) is crucial, as it provides a foundational reference for understanding canine mammary neoplasms. The relation between the Proliferation Index (PI) indicated by Ki-67 expression, along with the Apoptotic Index (AI) determined through Caspase-3 expression during the oestrous cycle, is inadequately documented in existing literature. This study seeks to offer insights into the interplay between PI and AI in the CMG across oestrous cycle phases. An extensive investigation was conducted on a diverse case series of bitches (n = 18). Oestrous cycle stages were determined through vaginal cytology, histological examination of the reproductive tract and serum progesterone and oestradiol concentrations. The entire mammary chain was histologically examined, and proliferation and apoptosis were assessed via double immunohistochemistry employing anti-Ki-67 and Caspase-3 antibodies. PI and AI were evaluated through a systematic random sampling approach, counting a minimum of 200 cells for each cell type. There was a significantly higher PI during early dioestrus in all mammary gland components, with a greater proportion of positive cells observed in epithelial cells compared to stromal cells. The highest PI was detected in epithelial cells within the end buds. Significant differences were found in Ki-67 labelling across the cranial mammary glands. A positive and strong correlation was noted between progesterone concentration and PI in epithelial cells. The AI remained consistently low throughout the oestrous cycle, with few differences observed across histological components. Caspase-3 labelling displayed the highest positivity in caudal mammary pairs. A negative and moderate correlation was identified between progesterone concentration and AI in interlobular mesenchymal cells. This study highlights the influence of endocrine regulation on cell proliferation indices in mammary tissue, emphasizing the need to consider these hormonal variations in toxicopathological studies involving canine mammary gland.

Aqueous Ethanolic Extract of Inonotus obliquus Ameliorates Polycystic Ovarian Syndrome by Modulating Oxidative Stress and Downregulating TNF-α and Interleukin-6

Polycystic ovarian syndrome (PCOS) is a complex metabolic and endocrine disorder with multifactorial etiology and complex pathogenesis. These intrinsic physiological changes cause anovulation, infertility, and miscarriage in women and exacerbate their chances of becoming hyperlipidemic and diabetic. Inonotus obliquus has been used traditionally for infertility problems. The present study aimed to investigate the therapeutic effects of aqueous ethanolic extract of Inonotus obliquus (AEIO) in female rats with letrozole-induced PCOS, along with the determination of its possible mechanism. HPLC analysis revealed the presence of gallic acid, chlorogenic acid, rutin, p-coumaric, benzoic acid, quercetin, salicylic acid, and kaempferol. Thirty female albino rats were acquired and divided into two groups (5 + 25) to induce PCOS. Letrozole (1 mg/kg) was used to induce the disease for 7 weeks (25 rats) except for the normal control (5 rats). The disease was confirmed by vaginal smear cytology, weight gain, and endocrinopathy. After disease induction, rats were divided into five groups (five rats in each group; disease control, metformin 20 mg/kg, AEIO 100, 300, and 500 mg/kg). After completion of the study, the animals were euthanized under the influence of anesthesia (chloroform). Ovaries were removed for histopathology, the liver was evaluated for oxidative stress biomarkers, and blood samples were collected for biochemical evaluation. Ovarian histopathology showed an abnormal architecture with cystic follicles and abnormal granulosa cells. Interestingly, treatment with AEIO restored normal ovarian histology with primary, growing, and developing follicles. After conducting a hormonal analysis, it was found that the induction of PCOS led to a significant increase (p &lt; 0.001) in luteinizing hormone (LH), insulin, and testosterone levels, while the levels of follicle-stimulating hormone (FSH) decreased. However, treatment with AEIO at doses of 100–500 mg/kg restored these levels to normal. PCOS induction also resulted in oxidative stress and lipid peroxidation by significantly decreasing antioxidant enzymatic markers (SOD, CAT, and GSH) and increasing levels of lipid peroxidation enzymatic markers (MDA). AEIO restored the levels of superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH), and reduced the level of malondialdehyde (MDA). In conclusion, antioxidant phytochemicals (gallic acid, chlorogenic acid, rutin, p-coumaric, benzoic acid, quercetin, salicylic acid, and kaempferol) rich extract alleviated PCOS symptomatology through modulating oxidative stress markers and eliminating ovarian low-grade inflammation by downregulating the expression of NF-κB associated TNF-α and IL-6.