The direct and indirect effects of kisspeptin-54 on granulosa lutein cell function.

STUDY QUESTION: What are the in vivo and in vitro actions of kisspeptin-54 on the expression of genes involved in ovarian reproductive function, steroidogenesis and ovarian hyperstimulation syndrome (OHSS) in granulosa lutein (GL) cells when compared with traditional triggers of oocyte maturation? SUMMARY ANSWER: The use of kisspeptin-54 as an oocyte maturation trigger augmented expression of genes involved in ovarian steroidogenesis in human GL cells including, FSH receptor (FSHR), LH/hCG receptor (LHCGR), steroid acute regulatory protein (STAR), aromatase, estrogen receptors alpha and beta (ESR1, ESR2), 3-beta-hydroxysteroid dehydrogenase type 2 (3BHSD2) and inhibin A (INHBA), when compared to traditional maturation triggers, but did not alter markers of OHSS. WHAT IS KNOWN ALREADY: hCG is the most widely used trigger of oocyte maturation, but is associated with an increased risk of OHSS. The use of GnRH agonists to trigger oocyte maturation is a safer alternative to hCG. More recently, kisspeptin-54 has emerged as a novel therapeutic option that safely triggers oocyte maturation even in women at high risk of OHSS. Kisspeptin indirectly stimulates gonadotropin secretion by acting on hypothalamic GnRH neurons. Kisspeptin and its receptor are also expressed in the human ovary, but there is limited data on the direct action of kisspeptin on the ovary. STUDY DESIGN SIZE, DURATION: Forty-eight women undergoing IVF treatment for infertility consented to kisspeptin-54 triggering and/or granulosa cell collection and were included in the study. Twelve women received hCG, 12 received GnRH agonist and 24 received kisspeptin-54 to trigger oocyte maturation. In the kisspeptin-54 group, 12 received one injection of kisseptin-54 (9.6 nmol/kg) and 12 received two injections of kisspeptin-54 at a 10 h interval (9.6 nmol/kg × 2). PARTICIPANTS/MATERIALS, SETTING, METHODS: Follicular fluid was aspirated and pooled from follicles during the retrieval of oocytes for IVF/ICSI. GL cells were isolated and either RNA extracted immediately or cultured in vitro ± kisspeptin or hCG. MAIN RESULTS AND THE ROLE OF CHANCE: GL cells from women who had received kisspeptin-54 had a 14-fold and 8-fold higher gene expression of FSHR and a 2-fold (ns) and 2.5-fold (P < 0.05) higher expression of LHCGR than GL cells from women who had received hCG or GnRH agonist, respectively. CYP19A1 expression was 3.6-fold (P < 0.05) and 4.5-fold (P < 0.05) higher, STAR expression was 3.4-fold (P < 0.01) and 1.8-fold (P < 0.05) higher, HSD3B2 expression was 7.5- (P < 0.01) and 2.5-fold higher (P < 0.05), INHBA was 2.5-fold (P < 0.01) and 2.5-fold (P < 0.01) higher in GL cells from women who had received kisspeptin-54 than hCG or GnRHa, respectively. ESR1 (P < 0.05) and ESR2 (P < 0.05) both showed 3-fold higher expression in cells from kisspeptin treated than GnRHa treated women. Markers of vascular permeability and oocyte growth factors were unchanged (VEGFA, SERPINF1, CDH5, amphiregulin, epiregulin). Gene expression of kisspeptin receptor was unchanged. Whereas treating GL cells in vitro with hCG induced steroidogenic gene expression, kisspeptin-54 had no significant direct effects on either OHSS genes or steroidogenic genes. LIMITATIONS REASONS FOR CAUTION: Most women in the study had PCOS, which may limit applicability to other patient groups. For the analysis of the in vitro effects of kisspeptin-54, it is important to note that GL cells had already been exposed in vivo to an alternate maturation trigger. WIDER IMPLICATIONS OF THE FINDINGS: The profile of serum gonadotropins seen with kisspeptin administration compared to other triggers more closely resemble that of the natural cycle as compared with hCG. Thus, kisspeptin could potentially permit an ovarian environment augmented for steroidogenesis, in particular progesterone synthesis, which is required for embryo implantation. STUDY FUNDING/COMPETING INTEREST(S): Dr Owens is supported by an Imperial College London PhD Scholarship. Dr Abbara is supported by an National Institute of Health Research Academic Clinical Lectureship. The authors do not have any conflict of interest to declare. TRIAL REGISTRATION NUMBER: ClinicalTrials.gov NCT01667406.

Psychometric Properties of Persian Version of Structured Clinical Interview for DSM-5 for Personality Disorders.

OBJECTIVES: This study aims to examine the psychometric properties of the Persian version of the Structured Clinical Interview for DSM-5 for personality disorders (SCID-5-PD) among patients referred to psychiatric centres in Iran. METHODS: Between March 2017 and June 2019, 287 outpatients and inpatients aged 16 to 75 years who were referred to three psychiatric centres in Tehran, Iran were invited to participate. Patients were interviewed using the Persian version of the SCID-5-PD by two PhD students in clinical psychology who were blinded to patient records. Face validity and content validity of the Persian version of the SCID-5-PD were assessed by five specialists with ≥2 years of clinical experience. The agreement between the diagnoses made with the Persian version of the SCID-5-PD by the two PhD students in clinical psychology and the gold standard diagnoses made with DSM-5 by psychiatrists was determined, as were sensitivity, specificity, and positive and negative likelihood ratios. 109 (43.6%) patients were interviewed again after an interval of 7 to 10 days for inter-rater reliability and test-retest reliability. RESULTS: A total of 250 patients aged 17 to 74 (mean, 32.56) years were included. Face validity and content validity of the Persian version of SCID-5-PD were acceptable. The agreement between the Persian version of SCID-5-PD and DSM-5 (gold standard) was acceptable (kappa >0.4) for the diagnoses of obsessive-compulsive, paranoid, schizotypal, schizoid, histrionic, narcissistic, borderline, and antisocial personality disorders, whereas the agreement was unacceptable (kappa <0.4) for the diagnoses of avoidant and dependent personality disorders. Sensitivity for all diagnoses was high, except for avoidant (0.66) and dependent (0.66) personality disorders. Specificity for all diagnoses was high, except for avoidant personality disorder (0.66). The positive and negative likelihood ratios showed that the SCID-5-PD was accurate for diagnosing all personality disorders, except for schizoid personality disorder. Inter-rater reliability was good for all personality disorders, except for schizotypal personality disorder (0.531). Test-retest reliability was good for all personality disorders. CONCLUSION: The Persian version of the SCID-5-PD can be used to evaluate those who seek psychotherapy for all personality disorders, except for avoidant, dependent, schizoid, and schizotypal personality disorders.

Isolation and phenotypic and genotypic characterization of the potential probiotic strains of Lactobacillus from the Iranian population.

Among different causes of inflammatory bowel disease (IBD), the imbalance of the gut microbiome (dysbiosis) is one of the main reasons for the development of the disease. Probiotics are live microorganisms that can maintain gut microbiota by different mechanisms. We aimed to isolate and characterize the potential probiotic strains of Lactobacillus from the Iranian population. This cross-sectional study was conducted on faecal samples of 83 volunteer individuals living in Guilan Province, North Iran. The primary identification of Lactobacillus strains was performed by standard microbiological tests and confirmed by amplification of 16s rRNA specific primers. The acid and bile salt tolerance were assessed for all recovered strains. Also, the presence of 3 bacteriocins encoding genes was investigated by the PCR method. Totally, 42 samples were positive for Lactobacillus species. Acid and bile resistance assay showed that 67% and 33% of strains were resistant to acid and bile salt stress, respectively. Therefore, we found out that 28% of our Lactobacillus strains have the ability for resistance to acid and bile conditions. PCR results revealed that the prevalence of gassericin A, plantaricin S, lactacin bacteriocin genes were 16.6%, 12%, and 9.5%, respectively. Meanwhile, 5 out of 12 Lactobacillus strains that were resistant to acid and bile conditions contained one of the gassericin or plantaricin bacteriocins. We isolated 42 potential probiotic strains of Lactobacillus, of which the results of 5 strains were more promising and can be considered as potential probiotics sources for future functional products.

Prostaglandin F2-alpha receptor regulation in human uterine myocytes.

Investigations of the modulation of prostaglandin F(2alpha) receptor (FP) expression in primary cultures of human uterine myocytes showed that FP mRNA expression was reduced by progesterone, unaltered by cAMP (8-bromo cAMP or forskolin), but increased by the PKA antagonist H89. Interleukin (IL)-1beta, tumour necrosis factor-alpha and oxytocin increased FP mRNA expression and IL-6 and prostaglandin E(2) reduced FP mRNA expression. The changes in FP protein levels were similar to the mRNA responses. We found that the IL-1beta-induced increase in FP expression was mediated at least in part via protein kinase C (PKC), but was independent of mitogen-activated protein kinase, phospholipase C and PI3 kinase. Since IL-1beta activates NFkappaB, AP-1 and C/EBP, we over-expressed these transcription factors alone and in combination and found that only NFkappaB alone increased FP mRNA expression. Finally, we found that the IL-1beta-induced increase in FP expression was unaffected by progesterone and/or cAMP, but was accentuated by H89. These data suggest that the pregnancy-induced down-regulation in myometrial FP expression is mediated by progesterone and cAMP and that the increase with labour is induced by inflammatory cytokine activation of PKC and NFkappaB.

Modified protocol for improvement of differentiation potential of menstrual blood-derived stem cells into adipogenic lineage.

OBJECTIVES: To characterize potency of menstrual blood-derived stem cells (MenSCs) for future cell therapies, we examined differentiation potential of MenSCs into adipocytes. MATERIALS AND METHODS: Differentiation potential of MenSCs in comparison to bone marrow stem cells (BMSCs) was assessed in conventional culture medium. Differentiation potential of MenSCs into adipocytes was improved using different combinations of growth factors and hormones. RESULTS: First, we demonstrated that MenSCs preserve their appearance and karyotypic stability during passages. Although these cells express mesenchymal stem cells markers, they cannot simply be classified as mesenchymal stem cells due to expression of embryonic stem cells marker, OCT-4. Oil red O staining showed that differentiated MenSCs in conventional medium with/without retinoic acid (protocols 1 and 2) did not attain adipocyte characteristics, whereas differentiated BMSCs in conventional medium accumulated oil vacuoles typically. Nevertheless, real-time RT-PCR results showed that LPL gene expression was up-regulated in both protocols 1 and 2, whereas LEPR was up-regulated only in protocol 2 (fortified with retinoic acid). Surprisingly, protocol 3 (including rosiglitazone) had odd influence on mRNA expression of all genes (LEPR, LPL and PPAR-γ). Oil red O staining confirmed fat-producing ability of MenSCs under protocol 3. CONCLUSIONS: Presented data suggest an efficient differentiation protocol for in vitro production of MenSC-derived adipocytes. These cells are suggested to be an apt alternative to BMSCs for future stem cell therapy of soft tissue injuries.