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Background: Cryptosporidiosis causes high morbidity and mortality in children under 2 years of age globally. The lack of an appropriate animal model that mimics the pathogenesis of disease in humans has hampered the development and testing of potential therapeutic options. This study aimed to develop and validate an infant baboon infection model of cryptosporidiosis. Methods: Eighteen immunocompetent weaned infant baboons aged 12 to 16 months were used. The animals were n = 3 controls and three experimental groups of n = 5 animals each inoculated with Cryptosporidium parvum oocysts as follows: group 1: 2 × 104, group 2: 2 × 105, group 3: 2 × 106 followed by daily fecal sampling for oocyst evaluation. Blood sampling for immunological assay was done on the day of infection and weekly thereafter until the end of the experiment, followed by necropsy and histopathology. Statistical analysis was performed using R, SPSS, and GraphPad Prism software. Analysis of variance (ANOVA) and Bonferroni post hoc tests were used for comparison of the means, with p < 0.05 considered as a significant difference. Correlation coefficient and probit analysis were also performed. Results: In all experimental animals but not controls, the onset of oocyst shedding occurred between days 2 and 4, with the highest oocyst shedding occurring between days 6 and 28. Histological analysis revealed parasite establishment only in infected animals. Levels of cytokines (TNF-α, IFN-γ, and IL-10) increased significantly in experimental groups compared to controls. Conclusion: For developing a reproducible infant baboon model, 2 × 104 oocysts were an effective minimum quantifiable experimental infection dose. [ABSTRACT FROM AUTHOR]

Endometriosis is a common gynecological disorder affecting approximately 10% of women of reproductive age who often experience chronic pelvic pain and infertility. Laparoscopy, which is invasive and expensive, is the gold standard for diagnosis of endometriosis. A simple minimallyinvasive test for endometriosis-specific biomarkers which is yet to be realized would offer a timely and accurate diagnosis for the disease thereby allowing early treatment intervention. Although aberrant microRNA expression has been implicated in endometriosis in several studies, conflicting results have been reported. This study hypothesized that the use of an appropriate animal model will provide a unique entry point for the discovery of biomarkers for early diagnosis of endometriosis. The study aimed at identifying miRNAs that are differentially expressed in eutopic endometrium of induced endometriosis in Papio anubis for early detection of endometriosis. Female adult baboons (n = 3) were induced with endometriosis by intraperitoneal inoculation of autologous menstrual endometrium. We sequenced small RNA samples obtained from normal (control) and diseased eutopic endometrium. Quality reads from the sequences were subjected to differential expression analysis to identify dysregulated microRNAs and genes from other non-coding small RNA in the samples using a bioinformatics approach. Through in silico analysis, gene targets of the dysregulated miRNA and their functions were determined. Our findings show significant high expression of seven microRNAs namely miR-199a-3p, miR-145-5p, miR-214-3p, miR-143-3p, miR-125b-5p, miR-199a-5p and miR-10b-5p. The study also reveals five microRNAs that were significantly down regulated and they include miR-29b-3p, miR-16-5p, miR-342-3p, miR-378a-3p and let-7g-5p. Seventeen genes from non-coding small RNAs were significantly dysregulated. The dysregulated microRNAs and genes play important roles in pathogenesis of endometriosis. Our findings indicate that specific miRNA signatures are associated with endometriosis, and the dysregulated miRNAs could constitute new and informative biomarkers for early diagnosis of endometriosis. [ABSTRACT FROM AUTHOR]

Background: Ovarian tissue cryopreservation by vitrification is an attractive technique for fertility preservation in women. However, this technique has not been optimized. The aim of this study was to evaluate the baboon as a model for the preclinical study of ovarian tissue cryopreservation by vitrification and thawing. Methods: Ovarian cortical tissues (1-mm cubes) were obtained surgically from adult female olive baboons (n = 9) maintained in captivity and vitrified using dimethyl sulphoxide and ethylene glycol protocol. The proportion of morphologically intact follicles (primordial, primary and secondary) in paired fresh and cryopreserved (vitrified-thawed) ovarian tissues was compared. Results: Overall, 67.1% of follicles were morphologically normal after vitrification. When compared to fresh ovarian tissue, vitrified-thawed ovarian tissue contained a comparable number of intact primordial follicles (48.9 vs. 52.9%), and a lower number of both primary (14.8 vs. 29.5%; p < 0.05) and secondary (2.0 vs. 0.7%; p < 0.05) follicles. Conclusion: After vitrification and thawing, baboon ovarian tissue retains about 67% of morphologically normal follicles, which is comparable to results for human ovarian tissue, and suggests that the olive baboon is a promising animal model for preclinical assessment of ovarian vitrification, thawing and autotransplantation studies. Copyright © 2013 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

The biological effects of khat (Catha edulis) on reproduction and fertility are inadequately investigated and controversial hence we determined the effects of oral administration of high-dose khat on sperm parameters and male hormonal levels in olive baboons. In this study, 6 male baboons received a high dose of khat (500 g/week) during 1 month. Electroejaculation for sperm studies (concentration, motility and chromatin integrity) and plasma collection for hormonal analysis (testosterone, prolactin and cortisol) were done weekly during 1 month before and 1 month during khat administration as well as 2 weeks after the last dose of khat administration. Administration of khat extract induced a significant reduction in sperm motility (p = 0.008), sperm count (p = 0.041), sperm chromatin integrity (p = 0.0003), testosterone levels (p = 0.035) and prolactin levels (p = 0.0115), but not in cortisol levels and sperm volume (p > 0.05). The results suggest that high-dose khat decreases sperm quality and testosterone and hence may contribute to male infertility. [ABSTRACT FROM AUTHOR]

Background Development of a reproducible baboon in vitro fertilization ( IVF) system require optimized and reproducible sperm parameters. The objective of this study was to document basic spermatology values and investigate the reproducibility of these variables in the same baboons 1 or 3 months later in a larger number of baboons. Methods In this prospective study, sperm quality (semen volume, pH, concentration, motility, morphology and size) was evaluated in 27 sperm samples obtained from 9 baboons electroejaculated three times with a time interval of 1 month (between first and second sample collection) and 3 months (between second and third round sample collection). Results Baseline sperm values for semen volume (0.5 ± 0.3 ml), pH (7.5 ± 0.3), concentration (54.2 ± 19.3 million/ml), motility (67.3 ± 18.5%) and morphology (89.2 ± 4.8%) were similar to sperm samples obtained after 1 or 3 months ( P > 0.05). Head, midpiece and tail abnormalities were rarely observed (0-9%). Sperm dimensions were characterized by a tail length of 69.6 ± 13.9 μm, a head width of 2.41 ± 0.43 μm and a head length of 3.49 ± 0.6 μm. Conclusion Sperm quality was not affected by repeated electroejaculation with time intervals of 1 or 3 months, suggesting that the same baboon can participate multiple times in reproductive research. [ABSTRACT FROM AUTHOR]

Objective: A study was conducted to test the feasibility of cordocenteses and amniocenteses at different gestational ages in pregnant baboons. Study Design: Experiments were performed in 10 pregnant baboons at a median gestational age of 131 (range 92-169) days. At different time intervals, percutaneous samplings of amniotic fluid and fetal blood were performed under ultrasound guidance. Simultaneously, maternal blood samples were drawn. Results: With a median fetal weight of 431 g (range 111-690 g), 29 of 30 cordocenteses (96.6%) and all 30 amniocenteses and maternal samplings (100%) were successful in obtaining the required quantities for analysis. One cordocentesis was abandoned because of insufficient visualization of the umbilical cord due to a placental haemorrhage. Conclusion: Percutaneous amniocentesis and cordocentesis can be performed with a high success rate in the pregnant baboon model. In combination with a 100% success rate in obtaining simultaneous maternal blood samples, this method is able to provide data on transplacental transport. [ABSTRACT FROM AUTHOR]

Stress impacts nonhuman primate menstrual cycle length but the impact of quarantine is unknown. A retrospective analysis was performed on cycle data from 31 wild-caught baboons during and following quarantine. Cycling initiated in 94 days (19-181) and length normalized within 4-6 cycles. Quarantine significantly impacts menstrual cycle length. [ABSTRACT FROM AUTHOR]

Tests the hypothesis that eggs harvested from female baboons after ovarian stimulation with human gonadotrophins can be fertilized in vitro with baboon sperm and develop into embryos that can be transferred in the same baboon using a transcervical approach as in women. Materials and methods used in the study; Details of the process of ovarian stimulation and sperm collection; Result of the study.

Objective: To test the hypothesis that the c-Jun NH2-terminal kinase (JNK) inhibitor (JNKI) bentamapimod (AS602801/PGL5001) can reduce induced endometriosis in baboons.Design: Prospective randomized placebo-controlled study.Setting: Nonhuman primate research center.Animal(s): Twenty baboons each underwent four laparoscopies. Initial screening laparoscopy (L1) was followed after one rest cycle by an endometriosis-induction laparoscopy (L2). Fifty days after L2, the baboons were randomized just before staging laparoscopy (L3). Treatment lasted for 60 days, followed by a post-treatment staging laparoscopy (L4).Intervention(s): Randomization before a 60-day treatment in four groups: daily placebo (n = 5), daily oral administration of 20 mg/kg JNKI (n = 5), concomitant daily oral administration of 20 mg/kg JNKI and 10 mg medroxyprogesterone acetate (MPA; n = 5), or subcutaneous administration of 3 mg cetrorelix every 3 days (n = 5).Main Outcome Measure(s): Type, surface area and volume of endometriotic lesions, and revised American Society for Reproductive Medicine score and stage were recorded during L3 and L4. Menstrual cycle length and serum hormonal concentration were recorded before and after treatment.Result(s): Compared with placebo, treatment with JNKI, JNKI + PMA, or cetrorelix resulted in lower total surface area and volume of endometriotic lesions. Remodeling of red active lesions into white lesions was observed more frequently in baboons treated with JNKI + MPA than in baboons treated with JNKI only. Menstrual cycle length and serum hormonal concentration were similar between placebo and JNKI groups.Conclusion(s): JNKI alone was as effective as JNKI + MPA or cetrorelix in reducing induced endometriosis in baboons, but without severe side effects or effect on cycle length or serum reproductive hormones. [ABSTRACT FROM AUTHOR]

Objective: Hormonal suppressive therapy is not effective for endometriosis-associated subfertility and can even prevent conception. Medical inhibition of TNFα, which has been shown to improve conception, is effective in the prevention and treatment of endometriosis in baboons. Design: Prospective, placebo-controlled fertility trial. Setting: Animal research and laboratory facility. Animal(s): Sixteen adult female baboons with induced endometriosis. Intervention(s): All animals received a single IV dose of the anti-TNFα monoclonal antibody c5N (n = 9) or placebo (n = 7) at four different time points. The animals were then exposed to timed mating up to nine completed cycles or until pregnancy was achieved. Main Outcome Measure(s): Pregnancy rate (PR), cycle fecundity rate (CFR), time to pregnancy (TTP), and cumulative pregnancy rate (CPR). Result(s): Inhibition of TNFα did not result in a significant improvement in PR (100% c5N vs. 86% placebo), CFR (18% c5N vs. 30% placebo), median TTP (5 cycles c5N vs. 2 cycles placebo), or CPR (100% c5N vs. 80% placebo). The duration of the menstrual cycle was unchanged in both groups before and after the study. Two nonpregnant baboons in the c5N-group died during the study. Conclusion(s): Medical inhibition of TNFα allowed for normal conception but did not improve fecundity in baboons with induced endometriosis when compared with placebo. Larger studies with clinically available TNFα blockers in baboons with moderate to severe endometriosis are needed to further test the potential of these agents in the prevention or treatment of endometriosis-associated subfertility. [Copyright &y& Elsevier]

Endometriosis is a gynecological disorder affecting about 10% of women and can lead to invalidating painful symptoms and infertility. Since there is no current definitive cure for this disease, new therapeutic options are necessary. 17β-Hydroxysteroid dehydrogenase type 1 (17β-HSD1) is involved in the production of estradiol (E2), the most potent estrogen in women, and of 5-androstene-3β,17β-diol (5-diol), a weaker estrogen than E2, but whose importance increases after menopause. 17β-HSD1 is therefore a pharmacological target of choice for the treatment of estrogen-dependent diseases such as endometriosis. We developed a targeted-covalent (irreversible) and non-estrogenic inhibitor of 17β-HSD1, a molecule named PBRM, and herein evaluated its efficiency for the treatment of endometriosis. In a cell-free assay containing estrone (E1), the natural substrate of 17β-HSD1, PBRM was able to block the formation of E2 in a collection of 50 human endometriosis lesions from a different clinical feature type, location, and phase. When given orally by gavage at 15 mg/kg to baboons, the resulting plasmatic concentration of PBRM was found to be sufficiently high (up to 125 ng/mL) for an efficacy study in a non-human primate (baboon) endometriosis model. After 2 months of treatment, the number of lesions/adhesions decreased in 60% of animals (3/5) in the PBRM-treated group, compared to the placebo group which showed an increase in the number of lesion/adhesions in 60% (3/5) of animals. Indeed, the total number of lesions/adhesions decreased in treated group (−6.5 or −19% when excluding one animal) while it increased in the control group receiving a placebo (+11%). Analysis of specific endometriotic lesions revealed that PBRM decreased the number of red lesions (−67%; 8/12) and white lesions (−35%; 11/31), but not of blue-black lesions. Similarly, PBRM decreased the surface area of dense adhesions and filmy adhesions, as compared to placebo. Also, PBRM treatment did not significantly affect the number of menstrual days. Finally, this targeted covalent inhibitor showed no adverse effects and no apparent toxicity for the duration of the treatment. These data indicate that 17β-HSD1 inhibitor PBRM is a promising candidate for therapy targeting endometriosis and supports the need of additional efforts toward clinical trials. • 17β-HSD1 inhibitor PBRM blocks the estradiol synthesis in human endometriosis lesions. • In a non-human primate endometriosis model, PBRM reduced the number of lesions. • As targeted covalent inhibitor, PBRM showed no adverse effects and no apparent toxicity. • PBRM treatment did not significantly affect the number of menstrual days. • PBRM is a promising candidate for therapy targeting endometriosis. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the mechanism of action of recombinant human tumor necrosis factor (TNF)-binding protein-1 by assessing differential expression of messenger RNA (mRNA) for cytokines, matrix metalloproteinases, and growth and adhesion factors in baboons. Design: Analysis of gene expression in a prospective randomized study. Setting: University Fertility Center. Animal(s): In the in vivo study, 14 baboons were randomly and subcutaneously (SC) treated with either phosphate-buffered saline (PBS), GnRH antagonist, or recombinant human TNF-binding protein-1 at the time of induction. In the ex vivo study, 4 baboons were treated by menstrual endometrium that had been incubated randomly with either PBS or recombinant human TNF-binding protein-1 before intrapelvic injection. Intervention(s): In the in vivo study, analysis of 11 endometrial and 10 endometriosis biopsies included either PBS (n = 5), GnRH antagonist (n = 8), or recombinant human TNF-binding protein-1 (n = 8). In the ex vivo study, 2 endometrial and 4 endometriosis biopsies were analyzed from 4 baboons. Main Outcome Measure(s): The mRNA expression of TNF-α, IL-8, IL-6, transforming growth factor-beta (TGF-β), vascular endothelial growth factor, intercellular adhesion molecule-1, matrix metalloproteinase-1, and regulated on activation, normal T-cell expressed and secreted were investigated using real-time reverse transcriptase–polymer chain reaction (PCR). Result(s): TGF-β mRNA expression was decreased in endometriotic lesions from baboons treated with recombinant human TNF-binding protein-1 when compared with the placebo group. Conclusion(s): Except TGF-β, mRNA expression of inflammatory cytokines and adhesion/growth factors is not affected in endometrial and endometriosis biopsies from baboons after induction of endometriosis combined with either systemic injection of recombinant human TNF-binding or GnRH antagonist or ex vivo treatment with recombinant human TNF-binding protein-1. Further studies are needed to elucidate the mode of action on how inhibition of TNF-α activity prevents the development of endometriosis. [Copyright &y& Elsevier]