Single nucleotide polymorphisms in the Anti-Müllerian hormone (AMH Ile(49)Ser) and Anti-Müllerian hormone type II receptor (AMHRII -482 A>G) as genetic markers in assisted reproduction technology.

PURPOSE: The aim of the study was to evaluate whether the presence Antimullerian hormone (AMH) and Antimullerian hormone type II receptor (AMHRII) single nucleotide polymorphisms (SNPs) Ile(49)Ser and -482A>G respectively are related to the assisted reproduction outcome. METHODS: A prospective cross-sectional observational study was conducted in order to assess the distribution of AMH and AMHRII SNPs in two cohorts, one of healthy women (N = 100) and the control group and the IVF/ICSI group (N = 151) consisted of women undergoing IVF/ICSI treatment for infertility. Furthermore, a prospective longitudinal observational study was performed on the latter group to assess possible associations of these SNPs with patients' characteristics and controlled ovarian stimulation (COS) and pregnancy outcome. RESULTS: Among non-carriers of the AMH (Ile(49)Ser) polymorphism, basal FSH levels were lower in those with more than two of previous IVF attempts and fertilization rate was statistically higher in those with peak serum E2 levels below 1500 pg/ml, whereas among non-carriers of the AMHRII (-482 A>G) polymorphism, number of follicles was higher in those with more than two previous IVF attempts and total dose of gonadotropins was lower in those with peak serum E2 levels above 1500 pg/ml. CONCLUSIONS: There was evidence that in specific subgroups of women undergoing IVF/ICSI, AMH and AMHRII SNPs may be related to patients' characteristics and controlled ovarian stimulation and pregnancy outcome and thus may provide a means for the prediction of ovarian response in specific subgroups of women entering an IVF/ICSI program.

A Survey in Hawaii for Parasitoids of Citrus Whiteflies (Hemiptera: Aleyrodidae), for Introduction into Greece.

Whitefly species of Aleurocanthus spiniferus (Quaintance) and A. woglumi Ashby (Hemiptera: Aleyrodidae) are serious pests of citrus and other important fruit crops. The problem of citrus has initiated the successful introduction of several natural enemies for biocontrol programs in Hawaii and many other countries. Here, we summarized the history of infestation and biocontrol efforts of the two whiteflies in Hawaii for possible parasitoid importation into Greece. Two Platygasteridae (Amitus hesperidum Silvestri, A. spiniferus (Brethes), and three Aphelinidae (Encarsia clypealis (Silvestri), E. smithi (Silvestri), E. perplexa Huang, and Polaszek) were released in Hawaii for biocontrol of the citrus whiteflies during the period 1974-1999. The aphelinid Cales noacki Howard, purposely released for Aleurothrixus flococcus (Maskell) in 1982, was also reported to attack other whiteflies, including Aleurocanthus species, on citrus. An additional aphelinid parasitoid, Encarsia nipponica Silvestri, native to Japan and China, was accidentally introduced and found to attack both citrus whiteflies on the islands. Since the colonization of introduced parasitoids in infested fields on four Hawaiian Islands, no survey has been conducted to evaluate their potential impact. We conducted two short surveys during September-November 2022 on the islands of Kauai, Hawaii, and Oahu to introduce the dominant parasitoids to Greece for the biocontrol of A. spiniferus. Results showed that the infestation level was very low on Kauai, Hawaii, and Oahu Islands, with a mean infestation level range of 1.4-3.1 on Hawaii and Oahu Islands, mostly on pummelo and sweet orange, with no detection on the island of Kauai. The dominant parasitoid was characterized as Encarsia perplexa, using molecular analysis. Its parasitism rates ranged from 0 to 28% on the island of Hawaii and 11 to 65% on the island of Oahu. Emerged parasitoids have been reared in Greece for evaluation. This was the first field survey of Hawaii since the introduction and release of citrus whitefly natural enemies.

LH receptor gene expression in cumulus cells in women entering an ART program.

PURPOSE: Luteinizing hormone (LH) exerts its actions through its receptor (LHR), which is mainly expressed in theca cells and to a lesser extent in oocytes, granulosa and cumulus cells. The aim of the present study was the investigation of a possible correlation between LHR gene and LHR splice variants expression in cumulus cells and ovarian response as well as ART outcome. METHODS: Forty patients undergoing ICSI treatment for male factor infertility underwent a long luteal GnRH-agonist downregulation protocol with a fixed 5-day rLH pre-treatment prior to rFSH stimulation and samples of cumulus cells were collected on the day of egg collection. RNA extraction and cDNA preparation was followed by LHR gene expression investigation through real-time PCR. Furthermore, cumulus cells were investigated for the detection of LHR splice variants using reverse transcription PCR. RESULTS: Concerning LHR expression in cumulus cells, a statistically significant negative association was observed with the duration of ovarian stimulation (odds ratio = 0.23, p = 0.012). Interestingly, 6 over 7 women who fell pregnant expressed at least two specific types of LHR splice variants (735 bp, 621 bp), while only 1 out of 19 women that did not express any splice variant achieved a pregnancy. CONCLUSIONS: Consequently, the present study provide a step towards a new role of LHR gene expression profiling as a biomarker in the prediction of ovarian response at least in terms of duration of stimulation and also a tentative role of LHR splice variants expression in the prediction of pregnancy success.

Electrophysiological responses of Philaenus spumarius and Neophilaenus campestris females to plant volatiles.

The spittlebugs Philaenus spumarius and Neophilaenus campestris (Hemiptera: Aphrophoridae) are xylem-feeder insects that have been identified as vectors of Xylella fastidiosa in Europe. In the present study, we aim to identify volatile organic compounds (VOCs) that may act as semiochemicals for these species. Using the dynamic headspace technique, we collected VOCs from Olea europaea L. and Polygala myrtifolia L., highly susceptible plant species to X. fastidiosa, Pinus halepensis Mill., a common plant where N. campestris is found during summer, and from host plant species that are used as cover crops or exist as natural vegetation in olive orchards, such as Cistus creticus L., Medicago sativa L., Cynodon dactylon (L.) Pers., Lolium arundinaceum (Schreb.) Darbysh., Apium graveolens L. and Petroselinum crispum (Mill.) Fuss. We tested the response of female antennae on those blends with Gas Chromatography-Electroantennographic Detection (GC-EAD). The chemical profile of C. creticus and P. halepensis was rich in terpenes, alcohols, aldehydes, and esters. In the O. europaea profile, the main compounds were terpenes. As for P. myrtifolia, the volatile profile consisted mostly of alkanes and their substitutes. In the volatile profile of M. sativa, C. dactylon and L. arundinaceum, common volatile compounds were detected. Petroselinum crispum and Apium graveolens chemical profiles were dominated by terpenes. Several compounds elicited a consistent response to the female antennae of both species. In total, 65 compounds elicited consistent EAD responses for P. spumarius and 16 compounds for N. campestris. α-pinene was found in all tested plants and elicited consistent EAD responses of P. spumarius in five plants. In addition, antennae of P. spumarius females responded to camphor, limonene, 4-methyl octane and sabinene. These compounds were found in the volatile profile of at least 5 out of 8 examined plant species. Behavioral bioassays using Y-tube olfactometry were performed on volatiles that elicited antennal responses during electrophysiological studies. Among the compounds tested in behavioral studies, namely (-)-α-pinene, (+)-α-pinene, sabinene, (-)-S-limonene and (1R)-(+)-camphor, only the last one elicited a significant attraction response by P. spumarius females. The results achieved shed light on the VOCs from selected host plant species of X. fastidiosa that are perceived by two important insect vectors and a non-host plant, P. crispum. The identification of semiochemicals for manipulating spittlebugs' behavior contribute to the development of efficient monitoring tools for X. fastidiosa vectors.

"hCG priming" effect in controlled ovarian stimulation through a long protocol.

BACKGROUND: Recently, it has been demonstrated that, in patients down-regulated by GnRH analogues (GnRHa), a short-term pre-treatment with recombinant LH (rLH), prior to recombinant FSH (rFSH) administration, increases the number of small antral follicle prior to FSH stimulation and the yield of normally fertilized embryos. However, no data exist in the literature regarding the potential beneficial effect of "hCG priming" in controlled ovarian hyperstimulation (COH) through a long GnRH-a protocol, which binds the same receptor (LH/hCGR), though it is a much more potent compared to LH. The primary aims of this study were to assess the effect of short-term pre-rFSH administration of hCG in women entering an ICSI treatment cycle on follicular development, quality of oocytes and early embryo development. The secondary endpoints were to record the effects on endometrial quality and pregnancy rate. METHODS: Patients with a history of at least one previous unsuccessful ICSI cycle were randomly assigned into two groups to receive treatment with either a long protocol with rFSH (control group) or a long protocol with rFSH and pre-treatment with hCG (hCG group). In particular, in the latter group, a fixed 7 days course of 200 IU/day hCG was administered as soon as pituitary desensitization was confirmed. RESULTS: The mean number of oocytes retrieved was not significantly different between the two treatment groups, although the percentage of mature oocytes tended to be higher but not significantly different in hCG-treated patients. The percentage of patients with more than one grade 3 embryos was higher in the pre-treatment group, which also showed a higher pregnancy rate. CONCLUSION: All the above clinical observations, in conjunction with previous data, suggest a point towards a beneficial "hCG priming" effect in controlled ovarian hyperstimulation through a long GnRH-a down-regulation protocol, particularly in patients with previous ART failures.

Vaginal evisceration after hysterectomy: a rare condition a gynecologist should be familiar with.

PURPOSE: To report a case of vaginal vault rupture with intestinal herniation per vagina after hysterectomy and highlight the risk factors, clinical presentation and treatment options of this rare gynecologic emergency. METHODS: A 70-year-old woman presented to the emergency department with vaginal evisceration, emerged 4 years after vaginal hysterectomy for uterine prolapse. Approximately 30 cm of the terminal ileum was irreducibly protruding through the vagina. RESULTS: The patient was transferred to the operating theatre. The prolapsed bowel was reduced via the combined vaginal-abdominal route and the vaginal cuff was closed with non-absorbable interrupted sutures. CONCLUSION: Awareness as well as high suspicion index among gynecologists and all involved care givers, is important for early diagnosis, given that vaginal evisceration is a potentially life-threatening condition necessitating prompt surgical intervention.

Effect of male mating history and age on remating by female European corn borer.

If mating with an inferior male has high fitness costs, females may try to avoid mating with these males. Alternatively, females may accept an inferior male to ensure they have obtained at least one mate, and/or to avoid the costs of resisting these males. We hypothesized that females compensate for mating with an inferior male by remating. We tested this hypothesis by measuring remating propensity in females that had mated with an old, multiply-mated male, a 9-day-old virgin male, or a young, virgin male. Females were more likely to remate when they had mated with multiply-mated males than when they had mated with a 9-day-old or young virgin male. We discuss the observed mating behavior by females in terms of sexual selection for multiple mating.

Myomas and Adenomyosis: Impact on Reproductive Outcome.

Among uterine structural abnormalities, myomas and adenomyosis represent two distinct, though frequently coexistent entities, with a remarkable prevalence in women of reproductive age. Various mechanisms have been proposed to explain the impact of each of them on reproductive outcome. In respect to myomas, current evidence implies that submucosal ones have an adverse effect on conception and early pregnancy. A similar effect yet is not quite clear and has been suggested for intramural myomas. Still, it seems reasonable that intramural myomas greater than 4 cm in diameter may negatively impair reproductive outcome. On the contrary, subserosal myomas do not seem to have a significant impact, if any, on reproduction. The presence of submucosal and/or large intramural myomas has also been linked to adverse pregnancy outcomes. In particular increased risk for miscarriage, fetal malpresentation, placenta previa, preterm birth, placenta abruption, postpartum hemorrhage, and cesarean section has been reported. With regard to adenomyosis, besides the tentative coexistence of adenomyosis and infertility, to date a causal relationship among these conditions has not been fully confirmed. Preterm birth and preterm premature rupture of membranes, uterine rupture, postpartum hemorrhage due to uterine atony, and ectopic pregnancy have all been reported in association with adenomyosis. Further research on the impact of adenomyosis on reproductive outcome is welcome.

Maternal-neonatal serum paraoxonase 1 activity in relation to the mode of delivery.

AIM: To investigate the effect of the mode of labour and delivery on the total antioxidant status (TAS) and the paraoxonase 1 (PON 1) serum activity in mothers and their newborns. SUBJECTS AND METHODS: One hundred six women with normal pregnancy were divided into 4 groups: group A (n = 28) with normal labour and vaginal delivery (VG), group B (n = 25) with scheduled caesarean section (CS), group C (n = 26) with "emergency" CS and group D (n = 27) with prolonged labour + VG. Blood was obtained from the mothers at the beginning of the labour process and immediately after delivery (pre- and post-delivery) as well as from the umbilical cord (CB). PON 1 activity and blood chemistry were determined using the Bayer Advia 1650 Clinical Chemistry System, whereas TAS levels were measured spectrophotometrically at 450 min in microtiter plates. RESULTS: TAS levels were similar pre-delivery and low in CB in all the groups. In contrast, TAS levels were remarkably reduced in group C and in group D post-delivery whereas they were nearly unchanged in group B and just lowered in group A, at the same time of study. PON 1 activity was practically unaltered in group A and group B pre- vs. post-delivery. Interestingly, the enzyme activity was remarkably decreased in group C (222 +/- 16 vs. 153 +/- 14 U/min/mL) and group D (216 +/- 16 vs. 135 +/- 15 U/min/mL, p < 0.001) as compared with those of the other groups at the same time of study. Additionally, PON 1 activity was higher in the newborns of group A and group B than those in group C and group D. TAS and HDL positively correlated with PON 1 activity. CONCLUSION: The low TAS levels and the decreased PON 1 activity, which were found in groups C and D post-delivery, may be due to the increased production of free radicals, during long-lasting labour + VG and obstructive labour + CS. PON 1 activity was low in CB irrespectively of the mode of delivery, probably due to the low lipid levels in the serum of the umbilical cord. Neonates born with normal delivery or scheduled CS are benefited with a higher antiatherogenic enzyme activity perinatally.

Microarray Analyses Reveal Marked Differences in Growth Factor and Receptor Expression Between 8-Cell Human Embryos and Pluripotent Stem Cells.

Previous microarray analyses of RNAs from 8-cell (8C) human embryos revealed a lack of cell cycle checkpoints and overexpression of core circadian oscillators and cell cycle drivers relative to pluripotent human stem cells [human embryonic stem cells/induced pluripotent stem (hES/iPS)] and fibroblasts, suggesting growth factor independence during early cleavage stages. To explore this possibility, we queried our combined microarray database for expression of 487 growth factors and receptors. Fifty-one gene elements were overdetected on the 8C arrays relative to hES/iPS cells, including 14 detected at least 80-fold higher, which annotated to multiple pathways: six cytokine family (CSF1R, IL2RG, IL3RA, IL4, IL17B, IL23R), four transforming growth factor beta (TGFB) family (BMP6, BMP15, GDF9, ENG), one fibroblast growth factor (FGF) family [FGF14(FH4)], one epidermal growth factor member (GAB1), plus CD36, and CLEC10A. 8C-specific gene elements were enriched (73%) for reported circadian-controlled genes in mouse tissues. High-level detection of CSF1R, ENG, IL23R, and IL3RA specifically on the 8C arrays suggests the embryo plays an active role in blocking immune rejection and is poised for trophectoderm development; robust detection of NRG1, GAB1, -2, GRB7, and FGF14(FHF4) indicates novel roles in early development in addition to their known roles in later development. Forty-four gene elements were underdetected on the 8C arrays, including 11 at least 80-fold under the pluripotent cells: two cytokines (IFITM1, TNFRSF8), five TGFBs (BMP7, LEFTY1, LEFTY2, TDGF1, TDGF3), two FGFs (FGF2, FGF receptor 1), plus ING5, and WNT6. The microarray detection patterns suggest that hES/iPS cells exhibit suppressed circadian competence, underexpression of early differentiation markers, and more robust expression of generic pluripotency genes, in keeping with an artificial state of continual uncommitted cell division. In contrast, gene expression patterns of the 8C embryo suggest that it is an independent circadian rhythm-competent equivalence group poised to signal its environment, defend against maternal immune rejection, and begin the rapid commitment events of early embryogenesis.

Expression profiling of vulvar carcinoma: clues for deranged extracellular matrix remodeling and effects on multiple signaling pathways combined with discrete patient subsets.

The molecular mechanisms of vulvar squamous cell carcinoma (VSCC) remain obscure. To this end, we investigated systematically for the first time the expression profile of VSCC using the microarray technology, in a total of 11 snap-frozen samples, from five VSCC patients covering early and advanced stages of VSCC undergoing radical surgery and from six matched healthy controls. All experiments were performed using Affymetrix Human Genome U133A 2.0 oligonucleotide arrays, covering 22,277 probe sets. Genes were filtered and analyzed using analysis of variance, t test, fold-change calculations, and unsupervised hierarchical cluster analysis. Further processing included functional analysis and overrepresentation calculations based on Gene Ontology, Database for Annotation, Visualization, and Integrated Discovery, and Ingenuity Pathway Analysis. The molecular phenotypes of VSCC patients exhibited significant and discrete transcriptional differences from the healthy controls, whereas principal component analysis documented that this separation is mediated by a consistent set of gene expression differences. We detected 1077 genes (306 upregulated and 771 downregulated) that were differentially expressed between VSCC patients and healthy controls by at least twofold (P < .01), whereas a novel subset of patients was revealed displaying a distinct pattern of 125 upregulated genes involved in multiple cellular processes. Functional analysis of the 1077 genes documented their involvement in more than 50 signaling pathways, such as PTEN, oncostatin M, and extracellular signal-regulated kinase signaling, affecting extracellular matrix remodeling and invasion. Comparison of our data set with those of the single VIN study revealed that the two entities share a limited number of genes and display unique features.

p16 INK4A is a strong biomarker for cervical intraepithelial neoplasia and invasive cervical carcinoma: a reappraisal.

To assess the role of p16( INK4A), bcl-2, and p53 in cervical cancer screening, we conducted a retrospective trial of prospectively collected data. Sixty two women with abnormal Pap smears were subjected to colposcopy and biopsies from any abnormal lesion. Human papillomavirus (HPV)-DNA typing, histology, and immunochemistry for p16(INK4A), bcl-2, and p53 were performed for these women. Histologic diagnosis was that of low grade squamous intraepithelial lesion (LSIL), high grade squamous intraepithelial lesion (HSIL), or cancer in all cases. Human papillomavirus strains were identified in 56 patients (90.3%). All HSIL and invasive cancer cases were HPV positive. p16(INK4A) immunostaining yielded 100% sensitivity, 76% specificity, 61% positive predictive value, and 100% negative predictive value in cancer patients. The corresponding performance indicators for HSIL patients were 75%, 62%, 32%, and 91%. bcl-2 and p53 expression did not correlate with worsening grades of cervical disease. We conclude that p16(INK4A) seems to be a sensitive biomarker of high grade cervical intraepithelial neoplasia and cancer.

The effect of the mode of delivery on the maternal-neonatal erythrocyte membrane acetylcholinesterase activity.

UNLABELLED: Free radical production and high catecholamine levels are implicated with the modulation of acetylcholinesterase (AChE) activity. OBJECTIVE: To investigate the effect of the mode of delivery on maternal-neonatal erythrocyte membrane AChE activity. SUBJECTS AND METHODS: Some women with normal pregnancy (N = 30) were divided into two groups: group A (N = 16) with normal labour and vaginal delivery and group B (N = 14) with scheduled Cesarean section, twenty non-pregnant women were the controls. Blood was obtained from controls and from mothers pre- vs post-delivery as well as from the umbilical cord (CB). Total antioxidant status (TAS), membrane AChE activities and catecholamine blood levels were measured with a commercial kit, spectrophotometrically and HPLC methods, respectively. RESULTS: TAS and catecholamine levels as well as membrane AChE activities were similar in the two groups of mothers pre-delivery and in controls. TAS levels were reduced whereas AChE activities and catecholamine levels were increased post-delivery in mothers of group A and unaltered in group B at the same times of study. AChE activity was similarly lower in the CB of neonates than those of their mothers pre-delivery. CONCLUSIONS: During a normal delivery process, the low TAS, the increased levels of catecholamines and the increased AChE activity, post-delivery, may be due to the increased stress due to the participation of uterus and skeletal muscles as during endurance exercise. The low AChE activity in newborns may be related to perinatal immaturity.

Maternal-neonatal 8-hydroxy-deoxyguanosine serum concentrations as an index of DNA oxidation in association with the mode of labour and delivery.

AIM: To investigate the effect of the mode of labour and delivery on the total antioxidant status (TAS), and the biomarker of DNA oxidation, 8-hydroxy-deoxyguanosine (8-OHdG) serum levels, in mothers and their newborns. SUBJECTS AND METHODS: Some 106 women with normal pregnancy and normal blood biochemical parameters were divided into 4 groups: Group A (n=28) with normal labour and vaginal delivery (VG), Group B (n=25) with scheduled cesarean section (CS), Group C (n=26) with 'emergency' CS, and Group D (n=27) with prolonged labour+VG. Blood was obtained from the mothers at the beginning of labour, and immediately after delivery (pre- and post-delivery), as well as from the umbilical cord (CB). TAS, 8-OHdG and creatine kinase (CK) were measured in the sera with appropriate methodology. RESULTS: TAS levels were almost similar in all the groups pre-delivery, and in CB irrespective of the mode of labour and delivery, and remarkably decreased in Groups C and D post-delivery. 8-OHdG levels in Group C (0.94+/-0.08 ng/ml) and Group D (0.98+/-0.08 ng/ml) were significantly higher than those in Group A (0.26+/-0.01 ng/ml, p<0.001) and Group B (0.28+/-0.07 ng/ml, p<0.001) post-delivery. 8-OHdG levels were low in CB, independent of the mode of labour. CK positively correlated with 8-OHdG (r=0.48, p<0.001), the latter negatively correlated with TAS (r=-0.53, p<0.01). CONCLUSIONS: The lowest TAS and the highest 8-OHdG levels were found in Groups C and D post-delivery, probably due to the long-term participation of the mothers' skeletal and uterus muscles, whereas 8-OHdG levels were low in CB irrespective of the mode of delivery, possibly as a consequence of the antioxidant action of the placenta and/or the low lipid levels in the serum of the umbilical cord.