Vaginal Cytologic Examination Research Articles

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.

Toxicity and human health assessment of an alcohol-to-jet (ATJ) synthetic kerosene developed under an international agreement with Sweden

ABSTRACT Alcohol-to-jet (ATJ) Synthetic Kerosene with Aromatics (SKA) fuels are produced by dehydration and refining of alcohol feed stocks. ATJ SKA fuel known as SB-8 was developed by Swedish Biofuels as a cooperative agreement between Sweden and AFRL/RQTF. SB-8 including standard additives was tested in a 90-day toxicity study with male and female Fischer 344 rats exposed to 0, 200, 700, or 2000 mg/m3 fuel in an aerosol/vapor mixture for 6 hr/day, 5 days/week. Aerosols represented 0.04 and 0.84% average fuel concentration in 700 or 2000 mg/m3 exposure groups. Examination of vaginal cytology and sperm parameters found no marked changes in reproductive health. Neurobehavioral effects were increased rearing activity (motor activity) and significantly decreased grooming (functional observational battery) in 2000 mg/m3 female rats. Hematological changes were limited to elevated platelet counts in 2000 mg/m3 exposed males. Minimal focal alveolar epithelial hyperplasia with increased number of alveolar macrophages was noted in some 2000 mg/m3 males and one female rat. Additional rats tested for genotoxicity by micronucleus (MN) formation did not detect bone marrow cell toxicity or alterations in number of MN; SB-8 was not clastogenic. Inhalation results were similar to effects reported for JP-8. Both JP-8 and SB fuels were moderately irritating under occlusive wrapped conditions but slightly irritating under semi-occlusion. Exposure to SB-8, alone or as 50:50 blend with petroleum-derived JP-8, is not likely to enhance adverse human health risks in the military workplace.

Clinico-haemato-biochemical and Pathological Alteration of Pyometra in Canines

Background: Pyometra is a pathological state of the compromised uterus of grown-up to old, diestrum bitches. Pyometra is easy to identify but can be more challenging when there is no vaginal discharge. In the present study, an attempt was made to find out the pathogen involvement and clinico-haemato-biochemical alteration in bitches affected with Pyometra. Methods: Twelve bitches with a history of inappetence, polydipsia, dirty vaginal discharge, occasional vomition irrespective of breeds and age groups were clinically examined and diagnosed as pyometra. The presumptive clinical diagnosis was based on case history, clinical signs, vaginal cytology, abdominal ultrasonography or radiography, gross examination of a pus-filled uterus after the ovariohysterectomy. Result: Vaginal examination revealed brown chocolate malodorous vaginal discharge with discolouration of the perivulval tissue. Abdominal ultrasound showed hyperplasia with echogenic material in the uterine wall along with distended uterine horns containing a large volume of cellular contents. Radiographic imaging revealed cranio-dorsal displacement of the small intestine and a twisted uniform tubular opacity with a distended uterus. The vaginal cytology revealed a large number of degenerated neutrophils, intermediate and parabasal cells. Escherichia coli were the major pathogen isolated from the pyometra uteri of bitches. The surface of the endometrium was columnar ciliated epithelium, with hyperplasia of subepithelial tissue and thick muscularis. The haemato-biochemical evaluation in pyometra infected bitches revealed decreased haemoglobin (Hb), packed cell volume (PCV), total erythrocytic count (TEC) and lymphocyte count indicating the microcytic hypochromic type of anaemia. Leucocytosis with a predominant absolute neutrophilia and lymphocytosis were consistently found in canine pyometra. There was a significantly elevated profile of blood urea nitrogen, creatinine, transaminases and alkaline phosphatase as well as total proteins observed in pyometric female dogs as compared to healthy ones which may have diagnostic and prognostic importance, which also indicated damage of vital organ caused by bacterial endotoxins. The haemato-biochemical indices correlating with imaging techniques, vaginal cytology and histopathological examination can serve as good prognostic markers in bitches with pyometra.

Effects of the Mouse Estrous Cycle on Neuropeptide S Receptor‐Mediated Behaviors

The neuropeptide S receptor (NPSR) is a G protein‐coupled receptor that has been implicated in various disease states, such as anxiety and post‐traumatic stress disorder, and has shown promise as a target for the treatment of anxiety. Central brain administration of Neuropeptide S (NPS) in mice produces anxiolytic‐like effects, hyperlocomotion, and memory enhancement. Since current anxiety drugs cause undesirable side effects, development of more efficacious drugs is important. However, research into this system has been conducted solely in male rodents, despite females showing a higher prevalence for anxiety disorders. The current study investigated whether NPS‐mediated behavioral phenotypes seen in males translate to females, and whether they are affected by estrous cycle stage. Female C57BL/6 mice were intracerebroventricularly (ICV) cannulated and underwent various behavioral paradigms. Estrous cycle stage was determined through examination of vaginal cytology. Consistent with findings in male mice, NPS‐treated females displayed hyperlocomotion. Additionally, NPS‐treated females displayed anxiolytic‐like behaviors, but preliminary data suggests this effect is contingent on different phases of the estrous cycle. These results indicate that the NPSR could be a viable pharmaceutical target to treat anxiety disorders, as the anxiolytic phenotype is present in both sexes. However, in females, these effects seem to be dependent on the stage of the estrous cycle.

Evaluation of Bovine Progesterone Semi-Quantitative Test Kit (Rapid p4) for Estimation of Blood Serum Progesterone Levels in Follicular Phase of Bitch

BACKGROUND: Detection of ovulation time using progesterone measurement is important to inseminate or breeding during the optimum time. OBJECTIVES: The aim of present study was evaluation of bovine progesterone semi-quantitative test kit for estimation of blood serum progesterone levels in bitch. METHODS: Five healthy intact anestrus bitches were used in the present study. Dogs were treated with cabergolin until onset of proestrus. Vaginal cytology, ultrasonography and blood sampling were performed from the 7th day of proestrus until two days after ovulation. Blood serum samples were divided as follows: one part to measure progesterone level by using RIA, and the other part was investigated by bovine progesterone semi-quantitative test kit. Ovarian changes were evaluated by ultrasonography and assessment of the estrus cycle was performed by vaginal cytology examination. RESULTS: Following Spearman analysis, significant positive correlation was observed between the semi-quantitative test kit and RIA results (r=0.916; p =0.00). The results of ROC and Kappa analysis suggested that the highest diagnostic accuracy of progesterone semi-quantitative test kit was observed in the blood serum progesterone levels of 2 and 5 ng/ml. The sensitivity and specificity of the used kit at the level of 5 ng/ml of progesterone were 88.5 and 93.7 %, respectively. The sensitivity and specificity of the used kit at the level of 10 ng/ml of progesterone were 82.4 and 96 % and at the level of 10 ng/ml were 82.4 and 96%, respectively. The observed scores during the late proestrus were 2-3 and just one dog showed score 4. During the estrus phase, in two dogs scores 4-5 and in three dogs scores 3-5 were revealed. CONCLUSIONS: Bovine progesterone semi-quantitative test kit is useful for estimation of estrus cycles in bitch. Scores of 4 and 5 of semi-quantitative test kit indicate ovulation time and proper breeding time.

Hubungan antara skor hasil bacaan heat detector dengan gambaran sitologi vagina sebagai parameter penentuan estrus pada kambing

This study aimed to determine the relationship between heat detector reading results and the feature of vaginal swab preparations as a parameter for determining estrus in Peranakan Etawa goats. Five healthy non-pregnant goats weighing 20-30 kg, were synchronized using PGF2α (7.5 mg Dinoprost, Enzaprost) intramuscularly twice with 11 days interval. Vaginal cytology examination began the day after the second PGF2α injection. The heat detector (Draminski®, Dramiński SA, Poland) is an estrus detecting device based on the amount of electric current flowing through the vaginal mucus. Spearman correlation analysis showed a relationship (p <0.05) between heat detector reading and estrus detection based on vaginal cytology. The heat detector readings were very significantly correlated positively (p <0.01) with the number of epithelial cells (coefficient correlation, r= 0.564), but negatively correlated with the number of cornified cells (r = –0.854), and also significantly correlated positively (p <0.05) to the number of leukocytes (r = 0.487). The estrus stages were significantly correlated positively (p <0.05) only to the number of leukocytes (r = 0.918). Likewise, the number of epithelial cells was highly correlated negatively (p <0.01) only to cornified cells (r = –0,643), and the number of cornified cells was significantly correlated negatively (p <0.05) only to the number of leukocytes (r = –0.472). It could be concluded that the features of the proestrus, estrus and metestrus stages based on the goat vaginal cytology was correlated with the reading results of the heat detector.

The Evaluation of Clinicopathological and Immunohistochemical Findings of Cervical Clear Cell Carcinoma in a Bitch

Background: Clear cell carcinoma (CCC) of the cervix is a rarely seen malignant tumor which is classified in adenocarcinomas of the endometrium. It is responsible for 2.15-3.0% of endometrial tumors. The diagnosis is difficult and is still controversial in uterine carcinomas. Macroscopically, masses were protruding to endometrial surface. However, the definitive diagnosis was carried by microscopical evaluation. The cells contain clear cytoplasm. The cells are seen in this pattern because there is either glycogenation or secretoric activity in the cells.Case: In the present case, clinicopathologic and immunohistochemical findings of clear cell carcinoma of the cervix were defined in a 3 year-old spayed Golden retriever bitch. Previously spayed bitch, which had vaginal discharge for the last month-long, was submitted to the Department of Obstetrics and Gynecology, Ankara University, Faculty of VeterinaryMedicine. In the general clinical examination, vaginoscopy was performed and vaginal cytology was obtained from smear. The source of discharge was detected as cervix uteri. In vaginal cytologic examination, erythrocyte, neutrophile and superficial cells were observed. In abdominal ultrasonography, a hypoechoic mass was detected at cranial part of the urinary bladder. To evaluate the general health condition of the dog, total blood counting and serum biochemistry were analyzed in addition to assessing its hormone prophlye. Estrogen and progesterone levels were evaluated. Estradiol (E2) level was measured as 23 pg/mL and progesterone level was measured as 1.96 ng/mL from collected serum. The cervical mass in diameters of 3x4x2.5 cm was removed in operation. In macroscopical examination, it had spherical and regularly shape.After the macroscopical examination, tissue samples were fixed in 10% buffered formalin. Then, the samples stained with Haematoxylin-Eosin (H&E), Masson’s trichrome and PAS stainings, respectively. At the macroscopic and histological examinations, the mass was diagnosed as clear cell carcinoma. Immunohistochemically, the tumor was stained with CK19 α-SMA, vimentin, desmin, TGF-β, VEGF, CEA, ER and PR markers. Other markers gave moderate to severe reactions in exception for no or weak ER and PR positivities. In direction of these results, clear cell carcinoma of cervix had been found remarkably due to first description in a spayed bitch on the basis of knowledge.Discussion: Clear cell carcinoma (CCC) is one type of the endometrial carcinomas. It is frequently known to have aggressive behavior and unfavorable prognosis. In human counterparts, clear cell carcinoma is frequently reoccurred in caudoabdominal and pelvic region even after being removed. The occurrence of clear cell carcinoma has highly increasedwith diethylstilbestrol usage during pregnancy. The situation on uterine carcinoma is nearly same in domestic animals. It is related to prolonged estrogenism. In this case, the bitch has been under prolong exposure to estrogenic effect. Indeed the estradiol level was high when compared to spayed female. Also, it is thought that the reason of high estradiol leveldoes not depend on the ovarian remnant. It is believed this situation may be relevant to exogen hormone usage. Vaginal smear confirmed erythrocyte (due to bleeding), predominantly superficial cells to it’s under estrogenic effect even despite being spayed.Keywords: clear cell carcinoma, cervix, clinicopathology, marker, bitch.

Diagnosis and management of vaginal dryness in menopause

Objective: To review the diagnosis and management of vaginal dryness in menopausal womenMethods: Literature reviewResults: Lack of estrogen negatively impacts the vagina and the urinary tract. The vagina becomes dry (atrophic) and then causing pain during sexual intercourse. Vaginal atrophy can be diagnosed in the form of vaginal dryness (75%), painful sexual intercourse (38%), itching and discharge complaints. Sometimes the patient complained pain in urinating, frequent night urination, in-continence and recurrent urinary tract infections. Mucose of the cervix, vagina and vulva are thin. Vagina can bleed easi-ly. Diagnosis can also be made by measuring the pH of the vagina by using litmus paper and vaginal cytology. The management involves administration of local estrogen treatment using weak estrogen (E3) in the form of a cream. E3 cream does not need to be combined with progestogen. Other type of local hormonal therapy (TH) is DHEA cream. Giving a strong estrogen (E2) or equin estrogen should always be combined with progestogen to prevent endometrium hyperplasia, either administered locally or systemically. E3 cream is also safe in women with breast cancer who experienced vaginal atrophy.Conclusion: Lack of estrogen causes vaginal atrophy with symptoms of vaginal dryness. Vaginal atrophy also causes urinary problems. Diagnosis is based on symptoms, examination of vaginal pH and vaginal cytology. The management is by adminis-tering HT with estrogen. E3 creams is the most effective in relieving complaints caused by vaginal dryness and complaints caused by the bladder.

Effects of bilateral whole vessel ovarian ligation on dogs’ ovarian function and histopathology

The most significant postoperative complication of spaying is bleeding from ovarian pedicles. In this study, all of the ovarian vessels were bilaterally ligated; then, ovaries were stimulated by gonadotropins. In a clinical trial and sequential control experiment, five healthy mixed-breed mature female dogs were randomly selected and kept under the new situation for 3 weeks. All ovarian vessels of the dogs were double ligated, bilaterally. For 13 weeks, blood samples, vaginal cytology, and ultrasound examinations continued weekly. All dogs received eCG (200 IU/dog, IM, 5 days) and hCG (500 IU/case IM, 5th day) 1 month after ovarian ligation. After 4 weeks, all dogs were ovariohysterectomized, and ovarian specimens were submitted for histopathological evaluations. Estradiol 17β and progesterone were assayed with ELISA kits. Estradiol concentration decreased after ligation and continued to decrease to the end of study (during 10 weeks). Estradiol concentrations were different before ligation and after hormone stimulation and ovariohysterectomy (P < 0.01). Progesterone concentration decreased suddenly after ligation and had a plateau during the study. Progesterone concentrations were different before ligation period and after ligation and hormone stimulation periods (P < 0.03). An ultrasonographical evaluation showed normal ovarian structures before ligation and ovarian adhesions to bursa and ovarian hyperechogenicity without the presence of active structures. Bleeding, congestion, primordial and primary follicle necrosis, and some degree of fibrosis were observed in the histopathological evaluations. Bilateral whole vessel ovarian ligation could inactivate dogs’ ovarian function during 8 weeks. So, we suggest ligation of whole vessels of dogs’ ovaries as an alternative method for ovariectomy.

Ovary and vaginal epithelium dynamics during the estrous cycle in Dasyprocta prymnolopha Wagler, 1831: ultrasound and cytological examinations.

The objective of the present study was to monitor and characterize morphological alterations in ovaries of agouti (Dasyprocta prymnolopha), reared in captivity, by using abdominal ultrasonography. All animals underwent daily vaginal cytological examination to identify the current cycle phase. For each phase of the estrous cycle, ultrasound examinations were carried out to identify and describe the morphology of both ovaries. Topographic parameters in an ultrasound window were established to locate the ovaries. The agouti estrous cycle lasted an average of 29.94 ± 6.77 days. During vaginal cytology examinations, all cell types were identified, and each phase of the estrous cycle was established by cell counts. No significant alterations were observed in the assessed ovarian morphometry measurements. In 75% of the animals examined, ovarian follicle presence was observed in the proestrus phase.

Sensitivity of preputial and vaginal exfoliative cytological examination for diagnosis of canine leishmaniosis (Leishmania infantum)

Aim of the study was to examine sensitivity of preputial and vaginal exfoliative cytological examination as a noninvasive alternative to lymph node, spleen and bone marrow cytology, for detection of Leishmania infantum amastigotes in dogs with leishmaniosis, as, in previous studies, the protozoa have been observed in the penis and prepuce of male dogs and in the vagina of female dogs with leishmaniosis. In total, 20 male and 18 female dogs with confirmed leishmaniosis were included inthe study. Three cytology smears were prepared from different sites of the preputial cavity of males and one smear was prepared from the anterior vagina of females. Leishmania amastigotes were not observed in these samples after microscopic examination for 20 min at 1,000× magnification. Therefore, preputial and vaginal exfoliative cytology is not recommended for routine diagnosis of canine leishmaniosis.

The Relation of Body Temperature and Vaginal Cytology Examination in Time Artificial Insemination Rate Fat-tailed Sheep (Ovis Aries) in The District Sidoarjo East Java

Ewes fertility is one aspect that needs to maximum level. Reproductive performance in sheep livestock in Indonesia has become one of the main determinants of reproductive success. This study has been carried out of the body temperature measurement and sampling of vaginal cytology of 60 fat-tailed sheep before artificial insemination in Sidoarjo East Java. Body temperature measurement is done in the rectum of reported ewes in heat and artificial insemination requested can be determined by observing the condition of the vulva of ewes such as the presence or absence of mucus through the vagina, vulva swollen and reddened vulva mucosa. vaginal cytology sampling with a cotton swab inserted into the vulva ± 15cm of ewes that reported in heat then interpreted by a microscope. The result showed that ewe’s body temperature in 39.1oC-39.2oC has a significant correlation with vaginal cytology type on oestrous phase. Keyword: Ewes, body temperature, vaginal cytology, the district Sidoarjo East Java

Diagnosis and management of vaginal dryness in menopause

Objective: To review the diagnosis and management of vaginal dryness in menopausal womenMethods: Literature reviewResults: Lack of estrogen negatively impacts the vagina and the urinary tract. The vagina becomes dry (atrophic) and then causing pain during sexual intercourse. Vaginal atrophy can be diagnosed in the form of vaginal dryness (75%), painful sexual intercourse (38%), itching and discharge complaints. Sometimes the patient complained pain in urinating, frequent night urination, in-continence and recurrent urinary tract infections. Mucose of the cervix, vagina and vulva are thin. Vagina can bleed easi-ly. Diagnosis can also be made by measuring the pH of the vagina by using litmus paper and vaginal cytology. The management involves administration of local estrogen treatment using weak estrogen (E3) in the form of a cream. E3 cream does not need to be combined with progestogen. Other type of local hormonal therapy (TH) is DHEA cream. Giving a strong estrogen (E2) or equin estrogen should always be combined with progestogen to prevent endometrium hyperplasia, either administered locally or systemically. E3 cream is also safe in women with breast cancer who experienced vaginal atrophy.Conclusion: Lack of estrogen causes vaginal atrophy with symptoms of vaginal dryness. Vaginal atrophy also causes urinary problems. Diagnosis is based on symptoms, examination of vaginal pH and vaginal cytology. The management is by adminis-tering HT with estrogen. E3 creams is the most effective in relieving complaints caused by vaginal dryness and complaints caused by the bladder.

Mucometra in a bitch as a consequence of a serious professional mistake

An apathetic 14-year-old Hungarian Puli bitch was presented with a seven day history of vaginal discharge and reported ovariohysterectomy (OHE) conducted six years ago. Clinical examination showed elevated body temperature (39.5?C), enlarged abdomen, and vulvar swelling. Despite a reported OHE in the anamnesis, ultrasound examination of the abdomen demonstrated the presence of both ovaries and a large anechogenic zone resembling the uterus. Vaginal cytologic examination and hormonal analysis indicated that the bitch was in the oestrus phase of the cycle. After laparotomy, both intact ovaries and an enlarged uterus with marked adhesions and filled with liquid content were visualised. The cervix was situated in the caudal part of the abdominal cavity and connected to the uterus by only an adhesion stump. After extirpation, uterus and ovarian tissue samples were sent for histological examination. Histological findings of uterus and ovarian tissue samples indicated a diagnosis of mucometra. Obviously, OHE was not conducted professionally, and the procedure performed only prevented conception, but ovaries and uterus continued to be active and caused a serious health disorder after six years. When only a part of the ovaries or uterus is left after OHE, health complications can appear up to ten years later. Our case testifies that even if both ovaries and the entire uterus are left after OHE without communication with cervix and vagina, a bitch can live without noticeable health disorders up to six years. This underlines the importance of lymphatic drainage and resorption processes in the uterus as well as evacuation of uterus content through the vagina.

Follow-up for cervical cancer: a Program in Evidence-Based Care systematic review and clinical practice guideline update.

In 2009, the Program in Evidence-based Care (pebc) of Cancer Care Ontario published a guideline on the follow-up of cervical cancer. In 2014, the pebc undertook an update of the systematic review and clinical practice guideline for women in this target population. The literature from 2007 to August 2014 was searched using medline and embase [extended to 2000 for studies of human papillomavirus (hpv) dna testing]. Outcomes of interest were measures of survival, diagnostic accuracy, and quality of life. A working group evaluated the need for changes to the earlier guidelines and incorporated comments and feedback from internal and external reviewers. One systematic review and six individual studies were included. The working group concluded that the new evidence did not warrant changes to the 2009 recommendations, although hpv dna testing was added as a potentially more sensitive method of detecting recurrence in patients treated with radiotherapy. Comments from internal and external reviewers were incorporated. Follow-up care after primary treatment should be conducted and coordinated by a physician experienced in the surveillance of cancer patients. A reasonable follow-up strategy involves visits every 3-4 months within the first 2 years, and every 6-12 months during years 3-5. Visits should include a patient history and complete physical examination, with elicitation of relevant symptoms. Vaginal vault cytology examination should not be performed more frequently than annually. Combined positron-emission tomography and computed tomography, other imaging, and biomarker evaluation are not advocated; hpv dna testing could be useful as a method of detection of recurrence after radiotherapy. General recommendations for follow-up after 5 years are also provided.

Subchronic oral toxicity of evodia fruit powder in rats

Ethnopharmacological relevanceEvodia, a fruit from Evodia rutaecarpa, has been used in oriental medicine, and since its various pharmaceutical actions, including anti-cancer activity, have become known, evodia has been widely used as a dietary supplement. However, information regarding its toxicity is limited. Materials and methodsEvodia fruit from Evodia rutaecarpa (Juss.) Benth. var. officinalis (Dode) Huang (0, 25, 74, 222, 667, and 2000mg/kg) was administered orally five times per week for 13 weeks. Clinical signs, body weight, food consumption, hematology, serum chemistry, urinalysis, vaginal cytology, sperm morphology, organ weight, and gross and histopathological findings were evaluated. ResultsUrinary ketone body excretion was detected in males at 667 and 2000mg/kg and in females at 2000mg/kg. An increase in absolute/relative liver weight was observed in both sexes at 2000mg/kg. Although levels of serum alanine aminotransferase, glucose, total cholesterol, and triglycerides were significantly reduced in males and/or females at 200 and/or 667 and 2000mg/kg, all values were within normal ranges and were considered non-adverse. In addition, no treatment-related differences in body weight, food consumption, hematology, vaginal cytology, sperm morphology, or gross and histopathological examination were detected. ConclusionsThe subchronic no-observable-adverse-effect level for evodia fruit powder following oral administration in rats is greater than 2000mg/kg.

Prediction of Parturition Date in the Bitch and Queen

Assessment of gestational age and parturition date prediction is a challenging, yet common task in clinical management of the pregnant bitch or queen. Knowing the approximate parturition date is essential for a thorough pregnancy monitoring. Radiographic and ultrasonographic methods are suitable in bitches and queens. In the bitch, moreover, ovulation timing by means of LH or progesterone assay, determination of onset of vaginal dioestrus by means of examination of vaginal cytology, and recognition of impending parturition by monitoring prepartal progesterone levels and body temperature variation are practical methods. A combination of different methods increases the accuracy of parturition date prediction.

15: Programmed reproductive dysfunction in low birthweight (LBW) offspring

LBW offspring are at increased risk of programmed adult obesity, and obesity in women is associated with reproductive dysfunction. However, it is unknown whether the reproductive dysfunction is solely a result of obesity or attributable, in part, to a gestationally programmed mechanism. Food restriction (FR) in rat pregnancy results in LBW pups with hypoleptinemia. When nursed normally, FR females develop pre-pubescent hyperleptinemia and adult obesity. Leptin affects the hypothalamic–pituitary-gonadol (HPG) axis both centrally, influencing secretion of hypothalamic gonadotropin-releasing hormone (GnRH) and pituitary gonadotropins, and peripherally, regulating ovarian steroidogenesis. We hypothesized that programmed altered leptin impacts the HPG axis, leading to reproductive dysfunction in LBW female offspring. Control dams received ad libitum food, whereas study dams were 50% FR from pregnancy day 10 to 21. All pups were nursed by Control dams and weaned at 3 weeks to ad libitum feed. At 1 day of age, female hypothalamic protein expression of GnRH and leptin receptor (ObRb), and ovarian ObRb were analyzed by Western Blot. Data were normalized to β-actin. At 10 months of age, reproductive cycling was assessed by daily examination of vaginal cytology. At 1d of age, LBW pups showed significant downregulation of hypothalamic GnRH (0.5-fold). ObRb expression was significantly upregulated in both the hypothalamus (2-fold) and the ovary (1.6-fold). By 10 months of age, FR adult females exhibited early anovulation as evident by persistent estrus phase with absence of normal cycling. The results suggest a marked dysfunction in basal hypothalamic reproductive index in the LBW newborns. It is likely that the early reduced leptin levels in LBW newborns alter growth, maturation and function of HPG axis which later lead to reproductive dysfunction.

Interruption of the canine estrous cycle with a low and a high dose of the GnRH antagonist, acyline

To test the efficacy and clinical safety of a low and high dose of the GnRH antagonist, acyline, on estrous cycle interruption and anovulation in female dogs, 20 proestrous (<3 d) bitches were randomly assigned to one of the following pharmacological protocols (given sc): acyline 110 μg/kg (ACY-L; n = 6); acyline 330 μg/kg (ACY-H; n = 8); or placebo (PLACE, n = 6). The animals were monitored (clinical and vaginal cytology examinations) daily for 60 d. Blood samples for serum progesterone serum concentrations were collected 14 d after treatment to determine if ovulation had occurred. Appearance of side effects and days to the onset of the first spontaneous estrous cycle after treatment were also recorded. In both ACY groups, but not the PLACE group, estrous cycles were interrupted after treatment ( P < 0.05). The interval from treatment to estrus interruption in ACY-L and ACY-H groups was 3.0 ± 0.6 and 3.2 ± 0.2 d, respectively (LSM ± SEM; P > 0.05). In the PLACE bitches, physical, behavioral and cytological proestrus slowly progressed to estrus and diestrus. Ovulation was absent in all ACY, but not in PLACE bitches ( P < 0.05). None of the females manifested side effects related to the treatments ( P > 0.05). Spontaneous return to a normal estrous cycle during the study period occurred in all ACY (ACY-L 19.5 ± 2.7 d vs ACY-H 24.8 ± 2.0 d; P > 0.05), but in none of the PLACE bitches ( P < 0.05). In conclusion, acyline efficiently, safely and reversibly interrupted an early phase of the estrous cycle in bitches by preventing ovulation.

Expression of brain prolactin releasing peptide (PrRP) changes in the estrous cycle of female rats

Prolactin releasing peptide (PrRP) is a neuropeptide with 31 or 20 amino acid residues and regarded as a potent and specific stimulator of pituitary prolactin. PrRP immunoreactive (PrRP-ir) neurons and mRNA are found in medulla oblongata and hypothalamus and the fibers containing PrRP are widely distributed in rat brains. Therefore, it is postulated that PrRP might act as a neurohormone or a neurotransmitter as well as a neuromodulator in the brain. In the present study, we probed the expression of brain PrRP in the estrous cycle of female rats and the relationship between brain PrRP and GnRH. Female rats were divided into four groups: the diestrus, the proestrus, the estrus and the metaestrus, which were identified by the vaginal cytological examination. Immunohistochemistry, reverse transcriptase-polymerase chain reaction (RT-PCR) and immunofluorescent double labeling histochemistry combining confocal laser scanning microscope (CLSM) were used. The results showed that PrRP immunoreactive neurons in nucleus of solitary tract (NTS) and ventrolateral reticular nucleus (VLRN) in the proestrus were less than those in the diestrus, the estrus and the metaestrus. Similarly, the relative optical density of PrRP-ir fibers of the bed nucleus of stria terminalis (BST) in the proestrus was decreased compared with those in other three groups. However, the brain PrRPmRNA level was higher in the proestrus and estrus than those in the metaestrus and diestrus. We also observed the co-localization of GPR10-immunoreactive (GPR10-ir) and GnRH-immunoreactive (GnRH-ir) neurons in hypothalamic medial preoptic area (MPO). The present results provide morphological evidences that PrRP in the female rat brains might participate in the regulation of the rat estrous cycle at least in a direct way.