Increase In Number Of Follicles Research Articles

Impact of polyamine supplementation on GnRH expression, folliculogenesis, and puberty onset in young mice

BackgroundThe hypothalamic-pituitary-gonadal (HPG) axis is pivotal in regulating reproductive functions, with gonadotropin-releasing hormone (GnRH) acting as a central regulator. Recently, polyamines have been shown to regulate the HPG axis, including GnRH expression and ovarian biology in old and adult rodents. The present study firstly highlights the age-specific variation in the polyamine and their corresponding biosynthetic enzymes in the ovary during aging, and further, the study focuses on the effect of polyamines, putrescine, and agmatine, in young female mice. Method and resultImmunofluorescence analysis revealed age-related differences in the expression of ornithine decarboxylase 1 (ODC1), spermine (SPM), and spermidine (SPD) in the ovaries, with adult mice exhibiting significantly higher expression levels compared to young and old mice. Likewise, qPCR analysis showed the mRNA levels of Odc1, Spermidine synthase (Srm), and Spermine synthase (Sms) show a significant increase in adult ovaries, which is then followed by a significant decline in old age. Histological examination demonstrated morphological alterations in the ovaries with age, including decreased follicle numbers and increased stromal cells in old mice. Furthermore, treatment with putrescine, a polyamine, in young mice resulted in larger ovaries and increased follicle numbers compared to controls. Additionally, serum levels of gonadotropin-releasing hormone (GnRH) and progesterone (P4) were measured, showing elevated levels in polyamine-treated mice. GnRH mRNA expression also increased significantly. Gene expression analysis revealed upregulation of genes associated with folliculogenesis such as Fshr, Bmp15, Gdf9, Amh, Star, Hsdb3, and Plaur in the ovaries and onset of puberty such as Tac2, and Kiss1, and a decrease in Mkrn3 in the hypothalamus of polyamine-treated mice. ConclusionThis study investigates the effect of polyamines in young immature female mice, shedding light on their role in upregulating GnRH, and enhancing folliculogenesis. Overall, these findings suggest that polyamines play a crucial role in ovarian aging and HPG axis regulation, offering potential therapeutics to reinstate fertility in reproductively challenged individuals.

G-CSF-mobilized peripheral blood mononuclear cells combined with platelet-rich plasma restored the ovarian function of aged rats

BackgroundRegenerative medicine with peripheral blood mononuclear cell (PBMC) transplantation sheds light on the issue of premature ovarian insufficiency (POI). However, the efficiency of PBMC treatment in natural ovarian aging (NOA) remains unclear. MethodsThirteen-month-old female Sprague–Dawley (SD) rats were used to verify the NOA model. Seventy-two NOA rats were randomly divided into three groups: the NOA control group, PBMC group, and PBMC+platelet-rich plasma (PRP) group. PBMCs and PRP were transplanted by intraovarian injection. The effects on ovarian function and fertility were measured after transplantation. ResultsTransplantation of PBMCs could restore the normal estrous cycle, consistent with the recovery of serum sex hormone levels, increased follicle numbers at all stages, and restoration of fertility by facilitating pregnancy and live birth. Moreover, when combined with PRP injection, these effects were more significant. The male-specific SRY gene was detected in the ovary at all four time points, suggesting that PBMCs continuously survived and functioned in NOA rats. In addition, after PBMC treatment, the expression of angiogenesis-related and glycolysis-related markers in the ovaries was upregulated, which indicated that these effects were associated with angiogenesis and glycolysis. ConclusionsPBMC transplantation restores the ovarian functions and fertility of NOA rats, and PRP could enhance the efficiency. Increased ovarian vascularization, follicle production, and glycolysis are likely the major mechanisms.

Dendrobium officinale Polysaccharide (DOP) Promotes Hair Regrowth in Testosterone-Induced Bald Mice.

Androgenetic alopecia can affect up to 70% of males and 40% of females; however, certain therapeutic medications offer partial and transitory improvement but with major side effects. Dendrobium officinale polysaccharide (DOP) has been reported to improve androgen-related hair loss in mice, but the molecular mechanism remains unclear. To explore the effects of DOP on androgenetic alopecia. In this study, testosterone was subcutaneously administered to shave dorsa skin of mice to establish androgenetic alopecia; the effects of DOP in androgenetic alopecia were explored by DOP administration. Testosterone treatment extended the time of skin growing dark and hair growing, decreased the mean numbers of follicles in skin tissues, decreased β-catenin and cyclin D1 levels, and elevated testosterone, DHT (dihydrotestosterone), and 5α-reductase levels. In contrast, DOP administration shortened skin growing dark and hair growing times, promoted follicle cell proliferation, increased follicle numbers, increased β-catenin and cyclin D1 levels, and decreased testosterone, DHT, and 5α-reductase levels. DOP application significantly improved testosterone-induced hair follicle miniaturization and hair loss, possibly through affecting the Wnt signaling and hair follicle stem cell functions. This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Influence of genistein and diadizine on regularity of estrous cycle in cyclic female Wistar rat: interaction with estradiol receptors and vascular endothelial growth factor.

Isoflavones are estrogenic compounds that exist in soy, clover, and peanuts. They are selective estrogen receptor modulators. The study was planned to explain the interactions of isoflavones with estrogen receptors alpha (ERα), beta (ERβ), and vascular endothelial growth factor (VEGF) expressions in ovarian and uterine tissues during different stages of the estrous cycle of regular cyclic female Wistar rats. Thirty-two regular cyclic females were divided equally into control group: fed casein-based diet and isoflavones group: fed casein-based diet and gavaged 50 mg/kg/day soy isoflavones extract 40%. The regularity of estrus cycles was monitored. Final body weight (FBW), weight gain (BWG), and ovarian and uterine weights were estimated. Histopathology and immunohistochemistry for ERα, Erβ, and VEGF in ovarian and uterine tissues were performed. All females (100%, n = 16) in control group showed regularity in estrous cycle compared to 62.5% (n = 10) in isoflavones group. Estrus and diestrus phases revealed prolongation and shortening in isoflavones rats than control, respectively. Nonsignificant variation was noted in the duration of the whole cycle of both groups. FBW and BWG significantly decreased however, ovarian and uterine weights increased significantly in all estrous phases of isoflavones group than control. Histopathology demonstrated an increase in number of follicles/ovaries besides, hyperplasia and proliferation of luminal epithelium with hydropic degeneration in the isoflavones group. Also, uterine connective tissue stroma showed edema in the isoflavones group during all estrous phases. Immunostaining percentages of ERα, Erβ, and VEGF protein expression were significantly elevated in the isoflavones group during all estrous phases. Isoflavones induced irregularity of the estrous cycle that was encountered by increased and altered ERα, Erβ, and VEGF expressions in ovarian and uterine tissues.

A Single Gene-Mediated Perturbation of the Human Genome Leads to Malignant Transformation in a Murine Transgenic Model

We hypothesized that induction or activation of a gene that directly affects DNA integrity can provide a tool for ongoing genomic changes in a cell and may eventually lead to oncogenesis. In the human genome, apurinic/apyrimidinic (AP) sites are created by number of external and internal factors including exposure to chemicals, UV radiation, DNA damaging agents, reactive oxygen species (ROS), DNA glycosylases and/or spontaneous hydrolysis by other mechanisms. AP sites are pre-mutagenic lesions that can also affect normal DNA replication. An evolutionarily conserved mechanism “base excision repair (BER)” has been established to identify and repair such sites. As part of BER, AP nucleases cleave the phosphodiester backbone at 5' to the AP site, prior to its repair. Although APEX 1 and 2 are the nucleases required for this repair, their hyperactivity has been connected with increased DNA breaks. We have previously observed that over expression of either APEX 1 or APEX 2 leads to malignant transformation with number of genomic changes including copy number alterations, single nucleotide variants (SNVs) and structural changes.In this study, we investigated if transgenic overexpression of APEX2 in B cells in mice will cause tumorogenesis. We specifically overexpressed APEX2 in B cells by generating Tg,CD19Cre+/-,APEX2+/- mouse lines and used Tg,CD19Cre+/- mouse (siblings of trail group) as control. The expression of APEX2 was detected by immunohistochemical (IHC) staining, which confirmed specific expression of APEX2 in lymph nodes. We next evaluated impact of APEX2 expression on B cell genome. We isolated B220+ B cells from spleen of 8 weeks old mice, and identified both increased single strand break (SSB) and double strand breaks (DSB). After 45 weeks of age and at variable times thereafter, 40% (23/58) of Tg,CD19Cre+/-,APEX2+/- mice showed enlargement of the lymph nodes and spleen. Histology showed effacement of normal tissue architecture by a nodular or diffuse population of atypical lymphoid cells in interfollicular areas. A distinct pattern of B220+BCL6+CD5- B-cell lymphomas were identified after IHC staining, which resemble human diffuse large B-cell lymphoma (DLBCL). To investigate the changes at genomic level, we performed whole genome sequencing of tumor tissues from 3 mice as well as heart tissues (with lowest APEX2 expression) from the same mice to be used as controls. From single nucleotide varients (SNVs) calling, we observed average of 24,388 SNVs in the tumor with acquisition of new copy number changes and significant number of structural variants including average of 43 translocations, 51 deletions, 21 duplications and 24 inversion. Indel frequency was variable with average of 348. We observed mutational hotspots on chr6, chr12 and chr16. Variant allele frequency (VAF) data shows clonality of chr11 related mutations. We observed most frequent T>G, and then T>C mutations in all 3 samples. Interestingly, the mutational signature analysis showed signature 17 accounting for an average of 60% of mutations followed by signature 9. According to the mutational signature profile of different diseases, both signature 9 and 17 are observed in human B cell lymphoma.To understand the genesis of the malignant transformation, we scarified Tg,CD19Cre+/-,APEX2+/- mice and control Tg,CD19Cre+/- mouse at 25 weeks after birth to see the pre-malignant changes in lymphoid organs. We observed enlarged and increased follicle numbers in lymph nodes and in spleen in Tg,CD19Cre+/-,APEX2+/- mice compared to control. A single cell sequencing is underway to detect clonal changes at an early stage.Our results demonstrate a single DNA-repair related gene responsible for inducing genomic instability with malignant transformation providing a model to study oncogenesis as well as possible preventative measures. DisclosuresAnderson:Bristol-Myers Squibb: Membership on an entity's Board of Directors or advisory committees; C4 Therapeutics: Other: scientific founder; Gilead Sciences: Membership on an entity's Board of Directors or advisory committees; Oncopep: Other: scientific founder; MedImmune: Membership on an entity's Board of Directors or advisory committees; Millenium Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees.

Therapeutic Potential of Autologous Adipose Derived Mesenchymal Stem Cells in Human POI and Ovarian Aging

Background Women play an important role in the work setting. This leads them to put off their motherhood, sometimes preventing them from getting pregnant. Delaying pregnancy face women with low ovarian response, such as in Premature Ovarian Insufficiency (POI) or Ovarian Aging (OA). There is no current treatment, although there is evidence of improving ovarian function by inyecting mesenchymal stem cells (MSC). Materials and Methods Prospective, observational study of 17 women who attended Pronatal Clinic from 2019 to 2020. Each patient was registered in Assisted Reproductive Treatment (ART) and was enrolled in ovarian treatment with an autologous adipose tissue Mesenchymal Stem Cell (AD-MSCs) protocol. Three groups were assembled: 1) Control: AMH >1.2 ng/mL, without AD-MSCs, 2) POI/OA: female infertility due to POI/OA with AMH <1.2 ng/mL and 3) Amenorrhea: female infertility due to POI/OA with amenorrhea and AMH <1.2 ng/mL. Variables: Age, weight, height, serum AMH, endometrial thickness, follicular size and number on day 2 and 11 of the menstrual cycle, oocyte number, number of blastocysts and pregnancy rate. Results Between month 2 and 5, after AD-MSCs inyection, POI/OA group showed an increase in follicle number (2 to 9) and size (13.5 to 15.5 mm) on day 11 of the menstrual cycle, which resulted in a higher number of MII oocytes (2.6 to 4.2), and an increase in number of blastocysts (0 to 3) and endometrial thickness (8.6 to 9.4). Regarding the Amenorrhea group, a reboot in menstrual cycle was observed, although no further development of blastocyst was found. Conclusion The AD-MSCs inyection directly in the ovary allowed an increase in number of blastocysts and improved pregnancy rates in POI/OA patients.

71 Effect of follicle-stimulating hormone dose and circulating progesterone before ovum pickup and invitro embryo production in pregnant Holstein heifers

Superstimulation with exogenous FSH before ovum pickup for invitro embryo production (IVP) improves embryo production. However, there is limited information on different FSH dose regimens and the effect of progesterone (P4) concentrations on IVP. Thus, the objective of the present study was to evaluate the effect of FSH dose and circulating P4 on ovarian superstimulation responses, before ovum pickup (OPU) on IVP in pregnant heifers. Pregnant Holstein heifers (n=47) 19.5±0.3 months of age with moderate body condition score (3.3±0.03; scale 1 to 5) were randomly assigned to 2 treatment groups, p-FSH (Folltropin-V, Vetoquinol) 160mg (FSH160) or p-FSH 300mg (FSH300), in a crossover design. Two sessions of OPU were performed on each animal at 48 and 62 days of gestation, with a washout interval between sessions of 14 days. Dominant follicle removal (DFR) was utilised to synchronize follicular wave emergence, and FSH treatments were initiated 36h later. Heifers in both groups received a total of 4 FSH injections 12h apart as follows: FSH160 (48.0, 42.7, 37.3, and 32.0mg) or FSH300 (90.0, 80.0, 70.0, and 60.0mg). All heifers underwent OPU 40h after the last FSH treatment. Blood samples were collected for P4 quantitation at OPU and number of small (<6mm), medium (6–10mm), and large (>10mm) follicles were determined before OPU. Cumulus–oocyte complexes (COCs) recovered were processed and invitro embryo production was performed using standard procedures. Differences between treatment groups were evaluated using generalized linear mixed models, and data are presented as mean±s.e.m. (Table 1). Heifers treated with 300mg of p-FSH had a greater (P<0.05) number of medium-size follicles, total number of follicles, total COCs recovered, number of viable COCs, and number of blastocysts produced. In addition, heifers treated with 300mg of p-FSH had a greater cleavage rate (P=0.03). Circulating P4 was on average 10.1±0.4ng mL−1 and did not differ between gestation days (P=0.12). Furthermore, circulating P4 did not affect (P>0.10) any of the outcomes evaluated, nor was there an FSH dose by circulating P4 interaction (P>0.05). In conclusion, increasing the dose of p-FSH before OPU from 160 to 300mg resulted in an increase in the total number of follicles, number of oocytes, and number of blastocysts produced in pregnant donors without changes in blastocyst rate, regardless of progesterone concentrations at the time of OPU. Table 1. Ovum pickup and invitro embryo production in pregnant heifers treated with different doses of p-FSH Item FSH160 FSH300 P-value Small follicles (n) 5.0±0.6 3.5±0.4 0.02 Medium follicles (n) 14.4±0.8 18.3±0.9 0.0003 Large follicles (n) 2.9±0.3 2.5±0.3 0.28 Total follicles (n) 22.3±0.7 24.3±0.9 0.04 Total oocytes (n) 12.8±0.7 14.7±1.0 0.02 Recovery rate (%) 56.2±2.1 59.7±2.7 0.20 Viable oocytes (n) 11.4±0.7 13.3±1.0 0.02 Viable oocytes (%) 89.9±1.7 88.0±2.4 0.22 Cleavage rate (%) 63.8±4.3 71.5±3.7 0.03 Cleaved oocytes (n) 7.2±0.7 9.3±0.9 0.14 Blastocysts (n) 2.7±0.4 3.6±0.7 0.02 Blastocyst rate (%) 22.0±3.3 22.4±3.6 0.74

Elevated antimullerian hormone is due to increased follicle number in polycystic ovary syndrome compared to controls

Elevated antimullerian hormone is due to increased follicle number in polycystic ovary syndrome compared to controls

Correlation of Anti-Müllerian Hormone with Polycystic Ovarian Disease and its Relation with Age

Polycystic ovarian disease (PCOD) is a common endocrine disorder of reproductive-aged women. Serum Anti-Müllerian hormone (AMH) increases in PCOD women due to increased follicle numbers. Objectives: The aim was to evaluate ovarian reserve by measuring level of AMH among PCOD women. Prospective cohort study was performed on 62 ladies who aged 25 to 40 years and diagnosed by Rotterdam criteria as PCOD in Sulaimani Maternity Teaching Hospital/Outpatient Gynecological Clinic during May 2018 to May 2019. Consents were obtained from patients for their inclusion. Inclusion criteria were age of 25-40 years and diagnosis of PCOD. The patients’ ages, body mass index (BMI) and level of AMH were recorded. Sample size estimation was performed by "GPower 3.1" and "IBM SPSS Statistics version 25" was used for analysis of data. P-value of (?0.05) was considered significant. The mean±SD (standard deviation) of age of women were 31.7±5 years. The majority of participants were housewives from inside of Sulaimani City and most of them were diagnosed to have PCOD through clinical features and results of ultrasonography. The mean±SD of BMI and AMH were 26.2±2.8 kg/m2 (ranged from 21.3 to 33.2 kg/m2) and 4.76±3.51 ng/ml (ranged from 0.1-14.6 ng/ml) respectively. There was significant association between age of the participants and AMH and the association of BMI and AMH was not significant. The level of AMH is decreased with aging and it is higher among PCOD women due to increased follicle numbers. The level of AMH was not associated with BMI.

Chinese herbal decoction of Wenshen Yangxue formula improved fertility and pregnancy rate in mice through PI3K/Akt signaling.

Traditional Chinese medicine (TCM) is an effective management to infertility. The association between TCM-mediated fertility and inhibition of phosphatidylinositol-3-kinase (PI3K) would be investigated. Institute of Cancer Research mice were treated with three herbal decoctions, named Wenshen Yangxue formula, Wenshen formula, and Yangxue formula, plus with human gonadotropins. PI3K inhibitor wortmannin was administrated to half of mice. Some index such as body weight, fertility ability would be investigated. The expression of P13K/Akt signaling was detected by using Western blot analysis. No difference was observed in body weight among groups. Mice receiving the administration of human gonadotropins and herbal decoctions showed increased follicle numbers, percentage of fertilization, and promoted embryonic development. The treatment of Wenshen Yangxue formula decoction showed the highest efficiency, significant higher than Wenshen and Yangxue formulas. And increased the expression of p-PI3K and p-Akt proteins. These results suggested the herbal decoctions promoted the fertilization of mice, which was related to the charge of PI3K/Akt activation.

712 A small molecule modulator of the wnt pathway (SM04554) as a potential topical treatment for androgenetic alopecia (AGA)

SM04554, a novel, small molecule Wnt signaling modulator was evaluated in preclinical studies for hair follicle regeneration and hair growth as a potential treatment for AGA. Male CD1 and C57Bl/6 mice were depilated with Nair and treated with SM04554 or vehicle. Effects during telogen phase were measured using C57Bl/6 mice, shaved and treated starting on post-natal day (P) 49. Effects of various dosing regimens (treatment durations and on-off cycles) was evaluated in NIH-III nude mice and Hanford mini-pigs. Assessment of visual hair growth and histological follicle counts were performed at multiple timepoints in all studies, with classification of follicle types (vellus, indeterminate and terminal) in the mini-pig study. Activation of Wnt signaling, hair cycle and follicle proliferation were measured by immunostaining for β-catenin, versican and Ki-67. In CD1 mice, SM04554 induced hair growth with 4-days treatment, with a 2-fold increase (p < 0.05) in total hair-follicle counts, compared to vehicle. In depilated C57Bl/6 mice, SM04554 induced hair growth with 10 days of treatment. In C57Bl/6 mice shaved and treated from P49, hair growth was observed after 3 weeks of treatment with SM04554 compared to >5 weeks with vehicle. In NIH-III nude mice, an increase in the number of hair-follicles (2-fold, p < 0.05) was observed in SM04554-treated animals on day 14, with appearance of hair on day 28 and further increase with 42 days of treatment compared to no hair growth with vehicle. In mini-pigs, 42 days of SM04554 treatment showed a significant (p < 0.05) increase in vellus follicle numbers that was sustained until day 112, as compared to other treatment intervals and vehicle. SM04554 increased total and nuclear β-catenin, versican and Ki-67 specifically in hair follicles compared to vehicle, indicating localized follicular activation of Wnt signaling and proliferation, not generalized to the epidermis. In rodents and mini-pigs, SM04554 increased follicle numbers and hair growth compared to vehicle. SM04554 has potential as a topical therapy for AGA. Clinical studies are ongoing.

Mesenchymal stem cell-derived angiogenin promotes primodial follicle survival and angiogenesis in transplanted human ovarian tissue

BackgroundWe have recently reported that human bone marrow-derived mesenchymal stem cells (MSCs) facilitate angiogenesis and prevent follicle loss in xenografted human ovarian tissues. However, the mechanism underlying this effect remains to be elucidated. Thus, determining the paracrine profiles and identifying the key secreted factors in MSCs co-transplanted with ovarian grafts are essential for the future application of MSCs.MethodsIn this study, we used cytokine microarrays to identify differentially expressed proteins associated with angiogenesis in frozen-thawed ovarian tissues co-transplanted with MSCs. The function of specific secreted factors in MSCs co-transplanted with human ovarian tissues was studied via targeted blockade with short-hairpin RNAi and the use of monoclonal neutralizing antibodies.ResultsOur results showed that angiogenin (ANG) was one of the most robustly up-regulated proteins (among 42 protein we screened, 37 proteins were up-regulated). Notably, the targeted depletion of ANG with short-hairpin RNAi (shANG) or the addition of anti-ANG monoclonal neutralizing antibodies (ANG Ab) significantly reversed the MSC-stimulated angiogenesis, increased follicle numbers and protective effect on follicle apoptosis.ConclusionOur results indicate that ANG plays a critical role in regulating angiogenesis and follicle survival in xenografted human ovarian tissues. Our findings provide important insights into the molecular mechanism by which MSCs promote angiogenesis and follicle survival in transplanted ovarian tissues, thus providing a theoretical basis for their further application.

Ultrasonographic and Hormonal Evaluation of Clinically Suspected Cases of Polycystic Ovarian Syndrome

Polycystic ovarian syndrome (PCOS) is a physiological disorder that causes many negative effects involving a variety of systems in the body, such as the endocrine, metabolic, psychological, and reproductive systems. An individual may present with a variety of reproductive, metabolic, and psychological problems. Method: In this descriptive cross-sectional study, women of reproductive age group (15–35 yrs), presenting with infertility, irregular menstruation, acne, hirsuitism and obesity were included. Consecutive sample method and total 139 patients were included. Detailed history with specific emphasis on history of infertility, age, menstrual history (oligomenorrhoea; regular or irregular menses), and obesity was done. Serum FSH, LH, and LH:FSH ratio levels were estimated. TVS/ TA, increase in number of follicles in ovary (multiple follicles), and increase in ovarian volume were noted. Result: Out of the 139 patients studied at PRH, maximum patients (88.5%) had normal levels of Sr. LH. These normal levels were of follicular and luteal phase. Remaining 16 (11.5%) had increased levels of Sr. LH. Out of the 139 patients studied at PRH, majority (99.3%) of the patients had normal levels of Sr. FSH. These normal levels were of follicular and luteal phase. Only 1(0.7%) patient had decreased level of Sr. FSH. Of the 139 patients studied at PRH with evidence of PCOS, 58 (41.7%) patients had increased ratio of Sr. LH:Sr. FSH (i.e., >2:1). Remaining 81(58.3%) patients had normal ratio. In the present study of 139 patients, 65 patients had exclusive USG findings, 13 (9.4%) patients had exclusive hormonal evidence and 45 patients had combined USG and hormonal evidence. Overall 16 (11.5%) patients had neither of the evidence (although they had clinical features of PCOS). Conclusion: Hormonal evidence of PCOS was present. Of these serums, LH levels were raised in majority of the cases and also raised LH:FSH ratio despite the values of hormones being in normal range. Both, i.e., USG alone and USG combined hormonal, were good evaluators for clinically suspected cases of PCOS. KEYWORDS: Polycystic ovarian syndrome, Hormonal levels, USG findings, Serum, FSHLH PCOS hyperandrogenism

Mating alters the rate of development of ovarioles in the ladybird, Propylea dissecta (Coleoptera: Coccinellidae)

The influence of female mating status on ovarian development of the ladybird, Propylea dissecta (Mulsant) (Coleoptera: Coccinellidae), was investigated under laboratory conditions. We assessed the extent to which ovariole development was affected by mating and for that we initially created a base line by observing age specific ovariole development. Results show that the number of follicles in each ovariole increased with the age of both virgin and mated females up to the age of 3 days, thereafter, no increase in number of follicles was recorded. Ovariole width also increased with age in both virgin and mated females up to 4 days, thereafter, no increase in ovariole width was recorded. The ovariole width of mated females was significantly greater than that of virgin females. Egg maturation and the egg load started to increase at the age of 8 days in virgin females. Thereafter, it increased with increase in female age. While in mated females, immature eggs were recorded in their ovarioles from the age of 1 to 2 days. In mated females, however, the increase in the number of mature eggs per ovariole and egg load started when they were 3 days old. Egg load continuously increased with increasing female age.

Constitutively active Foxo3 in oocytes preserves ovarian reserve in mice

During female reproductive life, ovarian follicle reserve is reduced by maturation and atresia until menopause ensues. Foxo3 is required to maintain the ovarian reserve in mice. Here we show that overexpression of constitutively active FOXO3 can increase ovarian reproductive capacity in mice. We find increased follicle numbers and decreased gonadotropin levels in aging FOXO3-transgenic mice compared with wild-type littermates, suggesting maintenance of a greater ovarian reserve. Based on cumulative progeny in aging animals, we find 31-49% increased fertility in transgenic females. The gene expression profile of Foxo3-/- knockout ovaries appears older than that of wild-type littermates, and the transgene induces a younger-looking profile, restoring much of the wild-type transcriptome. This is the first gain-of-function model of augmented reproductive reserve in mice, thus emphasizing the role of Foxo3 as a guardian of the ovarian follicle pool in mammals and a potential determinant of the onset of menopause.

Mechanistic action of mesenchymal stem cell injection in the treatment of chemically induced ovarian failure in rabbits

No curative treatment is known for primary ovarian failure; however, mesenchymal stem cells (MSCs), through self-renewal and regeneration, push the trial to evaluate their role in the treatment of ovarian failure. The aim of this study was to explore the impact of MSCs on cyclophosphamide (CTX)-induced ovarian failure in rabbits and to clarify the mechanism(s) by which MSCs exert their action. Thirty-five adult female rabbits were injected with CTX to induce ovarian failure. Five rabbits were euthanized after the last injection of CTX for histological examination. The others (30 rabbits) were further subdivided into two groups: group 1 (ovarian failure group, 15 rabbits) received no treatment; group 2 (ovarian failure and MSC recipient group, 15 rabbits) received MSCs isolated from extracted bone marrow of male rabbits. A decrease of follicle-stimulating hormone and an increase of estrogen and vascular endothelial growth factor (VEGF) levels in the MSC recipient group versus the ovarian failure group were found. Weak caspase-3 expression and+ve proliferating cell nuclear antigen staining after MSC injection were detected. Cytological and histological examinations showed increased follicle numbers with apparent normal structure of ovarian follicles in the MSC recipient group. Moreover, Y chromosome-containing cells from male donors were present within the ovarian tissues in group 2. The current study suggests that intravenous injection of MSCs into rabbits with chemotherapy-induced ovarian damage improved ovarian function. MSCs accomplish this function by direct differentiation into specific cellular phenotypes and by secretion of VEGF, which influence the regeneration of the ovary.

Effects of the anti-thyroidal compound potassium-perchlorate on the thyroid system of the zebrafish

The increasing pollution of aquatic habitats with anthropogenic compounds has led to various test strategies to detect hazardous chemicals. However, information on effects of pollutants in the thyroid system in fish, which is essential for growth, development and parts of reproduction, is still scarce. Other vertebrate groups such as amphibians or mammals are well-studied; so the need for further knowledge especially in fish as a favored vertebrate model test organism is evident. Modified early life-stage tests were carried out with zebrafish exposed to the known thyroid inhibitor potassium perchlorate (0, 62.5, 125, 250, 500 and 5000μg/L) to identify adverse effects on the hypothalamic-pituitary–thyroid axis. Especially higher perchlorate concentrations led to conspicuous alterations in thyroidal tissue architecture and to effects in the pituitary. In the thyroid, severe hyperplasia at concentrations ≥500μg/L together with an increase in follicle number could be detected. The most sensitive endpoint was the colloid, which showed alterations at ≥250μg/L. The tinctorial properties and the texture of the colloid changed dramatically. Interestingly, effects on epithelial cell height were minor. The pituitary revealed significant proliferations of TSH-producing cells resulting in alterations in the ratio of adeno- to neurohypophysis. The liver as the main site of T4 deiodination showed severe glycogen depletion at concentrations ≥250μg/L. In summary, the thyroid system in zebrafish showed effects by perchlorate from concentrations ≥250μg/L, thus documenting a high sensitivity of the zebrafish thyroid gland for goitrogens. In the future, such distinct alterations could lead to a better understanding and identification of potential thyroid-disrupting chemicals.

The infusion of glucose in ewes during the luteal phase increases the number of follicles but reduces oestradiol production and some correlates of metabolic function in the large follicles

Short-term nutritional supplementation stimulates folliculogenesis in ewes probably by insulin-mediated actions of glucose in the follicle. The aim of this study was to determine the effect of glucose on follicle number and granulosa levels of Aromatase P450 and phosphorylated Akt and AMPK. Twelve Ile-de-France ewes were allocated to two groups; one ( n = 7) infused with saline and the other ( n = 5) with glucose (10 mM/h) for 72 h in the luteal phase. At the end of infusion, ovaries were collected and all follicles >1 mm in diameter were dissected to recover granulosa cells. Aromatase P450 and phosphorylated Akt and AMPK were analysed by Western blotting of granulosa cell lysates. Blood plasmas collected before and during the infusions were analysed for progesterone, oestradiol, LH, FSH, glucose, insulin and IGF-I. The infusion of glucose significantly increased follicle number but, significantly reduced Aromatase P450 and phosphorylated Akt and AMPK in granulosa cells. The circulating concentration of glucose rose significantly 3 h after the start of the glucose infusion and remained elevated until 27 h then fell; the circulating concentration of insulin rose significantly by 3 h and remained elevated. The circulating concentration of oestradiol fell significantly by 32 h and remained low; the circulating concentrations of LH and FSH were unaffected. These data show that short-term infusion of glucose stimulated follicular growth but decreased Aromatase P450 in granulosa cells. The reduced levels of phosphorylated Akt and AMPK suggest that the phosphatidylinositol 3-kinase pathway has been inhibited by high concentrations of glucose. These data also suggest that there may be functional cross-talk between FSH and insulin signalling in granulosa cells.

Proliferating Active Cells, Lymphocyte Subsets, and Dendritic Cells in Recurrent Tonsillitis

To investigate the causes of hypertrophy in recurrent tonsillitis with hypertrophy (RTTH). An immunohistochemical study of recurrent tonsillitis with or without hypertrophy and adenoid tissue. Academic medical center. The study population comprised 15 patients with RTTH, 15 patients with recurrent tonsillitis (RT), 9 patients with adenoid vegetation, and 6 controls. The following outcome measures were investigated: follicle number and follicular area and circumference; degree of papillary arrangement and keratin cyst in crypts; fibrosis; and density of S-100(+) cells, CD20(+) cells, CD45RO(+) cells, lymphocytes and plasma cells, and cyclin D(1)(+) cells in surface epithelium, crypt epithelium, extrafollicular area, and follicles. In the RTTH group, follicle number and follicular area and circumference, S-100(+) cell density in crypt and surface epithelium, and CD20(+) cell density in crypt epithelium were higher than in the RT group. The other measures were lower in the RTTH group than in the RT group. In patients with RTTH, the increase in follicle number and S-100(+) cell density in surface epithelium and decrease in cyclin D(1)(+) cell density in surface epithelium were significant. The number of CD45RO(+) cells was unchanged, while S-100(+) cell density increased in surface epithelium; however, in the RTTH group, CD20(+) cell density, together with cyclin D(1)(+) cell density, decreased in surface epithelium when compared with the RT group. Also, there was a 50% decrease in cyclin D(1)(+) cell density in follicles, but CD20(+) cell density decreased minimally in follicles. In the RTTH group, the increase or decrease in the number of cyclin D(1)-expressing cells was parallel with the increase or decrease in the number of CD20(+) cells in the areas without follicles. Tonsil hypertrophy occurred with follicular hyperplasia and hypertrophy. There is a deficiency of proliferating active cells in response to mitogenic stimulation in RTTH. New follicles might have formed with B cells supplied from other sites because of the deficiency of proliferating active cells.

Letrozole in combination with gonadotropins decreases cancellation rates and improves ongoing pregnancy rates per transfer in poor responders

To evaluate the impact of letrozole on follicular recruitment and outcomes in IVF patients with a history of known poor response. Case control study. 19 patients defined as poor responders based on previous IVF performance were treated with letrozole 5 mg/day on cycle days (CD) 2–6. Gonadotropin injections were started on CD3. Suppression, ovulation and egg retrieval were performed according to established protocols. Patients' previous cycles were used as controls. Cycles were divided into two groups: Letrozole with FSH/hMG protocol (19 cycles) and conventional FSH/hMG protocol (38 cycles). Data were evaluated using chi-square for categorical variables and student t test or Mann-Whitney for continuous variables. Statistical analysis was performed with STATA 9.1 and P value of < 0.05 was considered significant. Table summarizes the results. TableVariableConventional-IVF (Cycles = 38)Letrozole-IVF (Cycles = 19)P valueAge (yrs)36.7 ± 2.137.0 ± 1.90.58Cycle cancellation (%)57.9 (22/38)0 (0/19)<0.001FSH/hMG used (IUs)3722 ± 10574484 ± 14670.08E2 day of hCG (pg/mL)757 ± 486731 ± 4180.87Follicles ≥14 mm3.1 ± 2.15.8 ± 2.0<0.001Grade1–2 embryos1.69 ± 1.661.68 ± 2.350.53Embryos transferred2.44 ± 1.12.11 ± 1.10.29Clinical PR (%)7.9 (3/38)21.1 (4/19)0.15Ongoing PR (%)2.6 (1/38)15.8 (3/19)0.02Total SAB rate (%)66 (2/3)25 (1/4)0.27Implantation rate (%)7.69 (3/39)13.2 (5/38)0.42Data presented as mean ± SD. Open table in a new tab Data presented as mean ± SD. In poor responders the addition of letrozole to conventional IVF stimulation was associated with a statistically significant increase in number of follicles and a higher ongoing pregnancy rate. Additionally, the cancellation rate was significantly reduced using the same criteria for egg retrieval (at least 4 follicles ≥14 mm and an estradiol ≥500 pg/ml). Our data suggests a benefit of adding letrozole to FSH/hMG protocols for poor responders. We speculate that the accumulation of intra-ovarian androgens may locally increase FSH receptivity and potentiate IVF stimulation (Mitwally; Fertil Steril, 2002). These findings corroborate previous work that demonstrated similar benefits of letrozole in poor response protocols (Garcia-Velasco; Fertil Steril, 2005). A prospective randomized study is indicated to further define the potential beneficial effect of letrozole in ovarian stimulation protocols.