Search

To investigate the influence of the Single Nucleotide Polymorphisms (SNPs) TP53 rs1625895, TP73 rs3765730, MMP9 rs17576, and MTHFR rs868014 on ovarian reserve (OR) in infertile patients.A prospective cross-sectional study was carried out in 145 infertile women. The patients were divided into two groups according to ovarian reserve, characterized by association between AMH levels and AFC:After patient distribution, both groups were compared (LOR X NOR) regarding the genotypes of the SNPs TP53 T/C rs1625895, TP73 G/A rs3765730, MMP9 Gln/Arg rs17576, and MTHFR A/G rs868014.The frequency of the TP53-T/T genotype was greater in the LOR and the TP53-C/C genotype was more frequent in patients with NOR. This association was confirmed by the frequency of alleles, where the presence of the T allele was significantly higher in patients who exhibited LOR (P = 0.0003). The frequency of the TP73-G/G genotype and of the G allele was higher in the LOR group (P = 0.01). Considering the MMP9 gene, the frequency of the Gln/Gln genotype was higher in the LOR group. However, the Gln/Arg genotype and the Arg allele prevailed in the NOR group (P = 0.006). The frequency of the MTHFR-A/A genotype was higher in the LOR group, whereas that of the MTHFR-GG genotype was higher in the NOR group. The presence of allele A was significantly higher in the LOR group (P = 0.002). The regression analysis shows that patients who present the TP53-T/T, TP73-G/G, MMP9-Gln/Gln, and MTHFR-A/A genotypes are 3.6X, 3.1X, 3.2X, and 3.7X more likely of having LOR, respectively. In addition, the association of the TP53/TT + TP73/GG genotypes increased the chance of women being included in the LOR group in 5.7-fold.The genotypes TP53-T/T, TP73-G/G, MMP9-Gln/Gln, and MTHFR-A/A increase the chance of women to exhibit LOR. These polymorphisms could be useful as genetic markers of low ovarian reserve in infertile patients.

Objective: To determine whether administration of progesterone on the day of oocyte retrieval may reverse accumulation of fluid in the endometrial cavity. Methods: A total of 50 patients who underwent assisted reproductive technology (ART) cycles with endometrial cavity fluid (ECF) observed by ultrasound at the time of oocyte retrieval were included. Upon the identification of ECF, vaginal administration of natural progesterone was started. Two days later, the endometrial cavity was re-evaluated, and embryo transfer was performed in the absence of ECF. Results: ECF was absent two days after administration of vaginal progesterone in 47 of the 50 patients (94%). ECF persisted in 3 of the 50 patients (6%). The clinical pregnancy rate per transfer was 34.0%, and the implantation rate was 21.6%. Conclusion: Our data suggest that, in the presence of ECF, administration of intravaginal progesterone in ART cycles must be initiated on the day of follicle aspiration to reverse ECF and to avoid the deleterious effects of fluid on the blastocyst-endometrial interaction.

Sirtuin 5 (SIRT5) is a protein lysine deacylase enzyme that regulates diverse biology by hydrolyzing -N-carboxyacyllysine posttranslational modifications in the cell. Inhibition of SIRT5 has been linked to potential treatment of several cancers but potent compounds with activity in cells have been lacking. Here we developed mechanism-based inhibitors that incorporate isosteres of a carboxylic acid residue that is important for high-affinity binding to the enzyme active site. By masking of the tetrazole moiety of the most potent candidate from our initial SAR study, we achieved potent and cytoselective growth inhibition for the treatment of SIRT5-dependent leukemic cancer cell lines in culture. Thus, we provide an efficient, cellularly active small molecule that targets SIRT5, which can help elucidate its function and potential as a future drug target. This work shows that masked biosisosteres of carboxylic acids are viable chemical motifs for the development of inhibitors that target mitochondrial enzymes, which may have applications beyond the sirtuin field.

Objective To assess the relationship between human blastocyst chromosomal ploidy established by niPGT-A and increasing age. Methods This is a prospective multicenter study carried out by ten assisted reproduction centers after their embryologists acquired training and validated their results with the previous use of niPGT-A. A total of 94 couples with indication for niPGT-A due to increase maternal age, male factor, repeated implantation failures, recurrent abortion or because they requested niPGT-A were included in this study. The couples had no karyotype abnormalities. After ICSI, the embryos were cultured until blastocyst stage using one or two step culture systems, single or sequential media respectively, at 37°C in an atmosphere of 6-7% CO2 and 5-20% O2 incubators. On day 3, we re-evaluated cleavage embryos to complete cumulus cells removal. The embryos were then cultured in individual well, with 20µl of medium under oil until they reached blastocyst stage. The blastocysts were vitrified and stored in liquid nitrogen. After that, the spent blastocyst culture medium (20µl) was transferred to a PCR tube and sent for analysis in the genetic laboratory, where it was stored at -80°C until sequencing. A total of 243 samples of spent blastocyst culture medium were collected on the 5th/6th day. Cell-free DNA secreted on culture medium was amplified using NICS Sample Preparation Kit (Yikon Genomics), based on the MALBAC technology. After whole genome amplification, the DNA was measured using a Qubit 2.0 fluorometer and subjected to next generation sequencing (NGS) using Illumina MiSeq® platform. The data were analyzed using the ChromGo® software (Yikon Genomics). Results The mean age of the patients was 38±4.08 years with an interval of 20-44 years. The euploid was diagnosed in 36.4% (80/220) of cases, aneuploidy in 31.3% (69/220), and mosaicism in 32.3% (71/220; with ≥60% aneuploidy) of blastocysts. Mosaic values ranged from 29.8% to 33.8% in different age groups. Individually, the most frequent chromosomal abnormality was XXY (Klinefelter Syndrome) occurring in 18 cases, followed by chromosome 21 (trisomy/monosomy) in 8 cases. The niPGT-A data showed a ≥60% incidence of aneuploid cells in all cases of chromosomal mosaicism (n=71). Conclusion A high degree of mosaicism with aneuploidy cells was detected, and some hypotheses were suggested for this data (niPGT-A sensitivity in detecting the self-correction of chromosomal abnormalities phenomenon). However, it did not vary remarkably with age. On the other hand, euploidy levels had a negative correlation with age and aneuploidy levels had a positive relationship. This is the first report in the literature to relate chromosomal ploidy in blastocysts using niPGT-A and increasing patient age.

Recently, a new technology known as the Noninvasive Preimplantation Genetic Testing for Aneuploidy (niPGT-A) emerged, using cell-free DNA present in the spent culture media of human blastocysts. Unlike PGT-A, in which only trophectoderm cells are used, niPGT-A reflects the ploidy state of these cells and internal cell mass, suggesting that this new technology may be less prone to error, being more reliable than the invasive test. The aim of the present study was to report the first occurrence of childbirth following niPGT-A in Brazil.

OBJECTIVE This study aimed to evaluate the effects of male age on sperm DNA damage. METHODS This cross-sectional study included semen samples collected from 2,178 men seen at an infertility clinic. For DNA integrity analysis, the proportions of spermatozoa showing DNA fragmentation (TUNEL assay), abnormal chromatin packaging/underprotamination (chromomycin A3), abnormal mitochondrial membrane potential (MMP/MitoTracker Green), and apoptosis (annexin V) were recorded. For group comparisons, enrolled subjects were divided into three groups based on their ages: ≤35 years; 36-44 years; and ≥45 years. The associations between age and sperm parameters were assessed using Spearman's rank correlation coefficient. RESULTS Although aging did not affect sperm apoptosis (p>.05), sperm DNA fragmentation and MMP deteriorated significantly with age (p

The nautilid Eutrephoceras dekayi (Morton 1834) is relatively abundant in the lower Maastrichtian (Upper Cretaceous) Pierre Shale of east-central Montana. We analysed the morphology, size frequency distribution, and isotope composition of a large collection of 220 well-preserved specimens including hatchlings, juveniles and adults. The newly hatched shell is approximately 14 mm in diameter with a body chamber one-third whorl in angular length terminating in the nepionic constriction. Internally, the embryonic shell contains five septa. Juveniles are abundant and comprise two-thirds of the sample whereas sub-adults, defined by the incipient flattening of the venter, are rare. Adults comprise approximately one-third of the sample and average 100 mm in diameter. The co-occurrence of newly hatched shells, small juveniles and adults suggests that the eggs were laid in the same area in which the hatchlings developed. Based on the excellent preservation of the juveniles, we conclude that they did not float into the area after death, but lived in the region, implying that this area served as a nursery for young animals. The calculated temperatures of the embryonic shells are similar to those of the post-embryonic shells and generally range from 16 to 18 °C. Upon hatching, the nautilids probably followed a demersal mode of life and lived in well-oxygenated water ≤50 m deep. An examination of lethal injuries (puncture holes) suggests that all ontogenetic stages were equally vulnerable to predation. The proximity of the site to the Sheridan Delta suggests that the specimens were smothered by sudden pulses of sediment transported into the area by major storms.

Objective The aim of this study was to evaluate the influence of different periods of abstinence on conventional semen parameters as well as functional parameters in human semen, including mitochondrial function, chromatin packing and sperm DNA fragmentation. Methods We recruited a cohort of 2,458 men undergoing infertility investigation. Semen analyses were performed according to WHO guidelines/morphology-motile sperm organelle morphology examination/MSOME. For DNA integrity analysis, the percentages of DNA fragmentation (TUNEL), abnormal chromatin packaging/underprotamination (chromomycin A3/CMA3), abnormal mitochondrial membrane potential (MMP/MitoTracker Green), and apoptosis (annexin-V) were recorded. Associations between the sexual abstinence period and sperm parameters were assessed using Spearman correlation. For group comparisons, the subjects were categorized according to the sexual abstinence period (SAP) into three groups: SAP 5 days. Results The duration of abstinence had a statistically significant positive influence on sperm concentration and volume, the number of leukocytes and a statistically significant negative influence on sperm motility and vitality. The percentages of DNA fragmentation and MMP (mitochondrial damage) worsened with the increased duration of abstinence. The percentage of sperm protamination was statistically significantly increased with abstinence. Conclusion Increase in the sexual abstinence period influences sperm quality. This study reinforces the importance of the duration of ejaculatory abstinence on semen parameter variation. It highlights the deleterious effect of increased abstinence on DNA damage, which is most likely associated with ROS (mitochondrial damage/number of leukocytes). The increase in chromatin packaging can represent a protective feature for DNA.

Objective A variety of studies randomizing women/cycles or oocytes/embryos has been carried out to compare different culture media for culturing embryos up to cleavage or blastocyst stages showing controversial results. A recent systematic review suggested that data in the literature are insufficient to conclude the best culture medium for embryo quality, pregnancy and implantation. The objective of this study was to evaluate whether there is any difference between two commercial culture media regarding clinical outcomes after IMSI cycles. Methods A total of 120 patients, ≤39 years of age, undergoing ART treatment submitted to the IMSI program were prospectively broken down and randomized into two groups: Group I (Cook media) and Group II (Vitrolife media). Results Our data demonstrated that there was no difference using all the media from Cook or all the media from Vitrolife, for culturing embryos till day 2, in the bench incubator at low O2 concentration, in relation to fertilization, embryo quality, pregnancy and implantation rates (p>0.05). Conclusion Both culture media used, Cook medium and Vitrolife medium, for the IMSI procedure and for later embryo culture with transfer on the second day, are equally effective and can be used depending on the ease and availability of acquisition.

Certain gene polymorphisms are associated with implantation failure and pregnancy loss. Studies of leukaemia inhibitory factor (LIF) gene polymorphisms are scarce. The LIF single nucleotide polymorphism (SNP) thymine (T)/guanine (G) (rs929271) was studied in women to determine whether an association existed with pregnancy outcomes after intracytoplasmic sperm injection (ICSI); 411 women who underwent ICSI were recruited. DNA was extracted from the peripheral blood, and the LIF gene SNP T/G (rs929271) was genotyped using real-time polymerase chain reaction. Participants were divided into three groups according to their LIF genotype: T/T (n = 168), T/G (n = 202) and G/G (n = 41). All IVF and ICSI procedures were carried out under the same clinical and laboratory conditions. The ICSI cumulative results (from fresh plus frozen cycles) of each genotype group were analysed. The G/G genotype in women was associated with a higher implantation rate (T/T: 15.9%, T/G: 16.2%, G/G: 27.0%; P < 0.05), ongoing pregnancy rate/patient (T/T: 31.5%, T/G: 36.1%, G/G: 53.7%; P < 0.05) and ongoing pregnancy rate/transfer (T/T: 18.5%, T/G: 20.2%, G/G: 36.7%; P < 0.05). LIF SNP T/G (rs929271) seems to be a susceptibility biomarker capable of predicting implantation efficiency and pregnancy outcomes.

Objectives/Hypothesis Assess the role of combined antimicrobial and surgical therapy for difficult-to-treat nontuberculous mycobacteria (NTM) lesions of the head and neck in children. Study Design Retrospective cohort. Methods Retrospective review of pediatric head and neck NTM lesions at a tertiary children's hospital from 1999 to 2012. Results Seventy-one children were diagnosed with NTM lesions. Age of presentation ranged between 7 and 204 months of age. Most patients (62%) had multiple lesions. Treatments included incision and drainage, curettage, antibiotics, excision, and any combination of surgery and antibiotics. Upon initial presentation, the most common treatment was surgical excision alone (n = 34) with a high complication rate (50%). In 18 cases, patients were initially treated with a combination of antibiotics and surgical excision due to the extent or location of the lesion(s). Complication rate in these patients was also high (67%). The most common complications in surgically excised NTM lesions included temporary or persistent facial nerve dysfunction (24.6%), poor wound healing/scarring (10.8%), and Frey's syndrome (6.2%). Conclusions Surgical excision with or without medical therapy for NTM cervicofacial lymphadenitis in high-risk regions commonly resulted in marginal mandibular nerve dysfunction (24.6%). Postoperative facial nerve weakness generally resolved within a year. High rates of complications and a lack of proven best approaches suggest tailoring the approach to address the potential risks in that particular patient based on location and severity. Level of Evidence 4 Laryngoscope, 2015

Cerebral non-oxidative carbohydrate consumption may be driven by a ß2-adrenergic mechanism. This study tested whether the 46G > A (G16R) single nucleotide polymorphism of the ß2-adrenergic receptor gene (ADRB2) influences the metabolic and cerebrovascular responses to administration of adrenaline. Forty healthy Caucasian men were included from a group of genotyped individuals. Cardio- and cerebrovascular variables at baseline and during a 60-min adrenaline infusion (0.06 μg kg-1 min-1) were measured by Model flow, near-infrared spectroscopy and transcranial Doppler sonography. Blood samples were obtained from an artery and a retrograde catheter in the right internal jugular vein. The ADRB2 G16R variation had no effect on baseline arterial glucose, but during adrenaline infusion plasma glucose was up to 1.2 mM (CI95: 0.36-2.1, P < 0.026) higher in the Gly16 homozygotes compared with Arg16 homozygotes. The extrapolated steady-state levels of plasma glucose was 1.9mM (CI95: 1.0-2.9, PNLME < 0.0026) higher in the Gly16 homozygotes compared with Arg16 homozygotes. There was no change in the cerebral oxygen glucose index and the oxygen carbohydrate index during adrenaline infusion and the two indexes were not affected by G16R polymorphism. No difference between genotype groups was found in cardiac output at baseline or during adrenaline infusion. The metabolic response of glucose during adrenergic stimulation with adrenaline is associated to the G16R polymorphism of ADRB2, although without effect on cerebral metabolism. The differences in adrenaline-induced blood glucose increase between genotypes suggest an elevated ß2-adrenergic response in the Gly16 homozygotes with increased adrenaline-induced glycolysis compared to Arg16 homozygotes.

The γ-aminobutyric acid (GABA) transporters (GATs) are key membrane transporter proteins involved in the termination of GABAergic signaling at synapses in the mammalian brain and proposed drug targets in neurological disorders such as epilepsy. To date, four different GAT subtypes have been identified: GAT1, GAT2, GAT3 and the betaine/GABA transporter 1 (BGT1). Owing to the lack of potent and subtype selective inhibitors of the non-GAT1 GABA transporters, the physiological role and therapeutic potential of these transporters remain to be fully understood. Based on bioisosteric replacement of the amino group in β-alanine or GABA, a series of compounds was generated, and their pharmacological activity assessed at human GAT subtypes. Using a cell-based [(3)H]GABA uptake assay, several selective inhibitors at human BGT1 were identified. The guanidine-containing compound 9 (2-amino-1,4,5,6-tetrahydropyrimidine-5-carboxylic acid hydrochloride) displayed more than 250 times greater potency than the parent compound β-alanine at BGT1 and is thus the most potent inhibitor reported to date for this subtype (IC50 value of 2.5 µM). In addition, compound 9 displayed about 400, 16 and 40 times lower inhibitory potency at GAT1, GAT2 and GAT3, respectively. Compound 9 was shown to be a substrate for BGT1 and to have an overall similar pharmacological profile at the mouse orthologue. Compound 9 constitutes an interesting pharmacological tool for specifically investigating the cellular pharmacology of BGT1 and is the first small-molecule substrate identified with such a high selectivity for BGT1 over the three other GAT subtypes.

With a growing demand for high-value crops such as nuts, fruits and vegetables, both the importance of insect pollinators ensuring the production of these and the public awareness of the need for pollination are growing. While other species like bumblebees or leafcutter bees can be of value in specific cropping situations, the European or Western honeybee, Apis mellifera, is the only insect pollinator which can be provided at scale to commercial producers. This is driving the development of large-scale pollination providers and migratory beekeeping strategies, putting considerable stress on the bees themselves. Simultaneously, honey-focused industries are emerging in countries with strong monofloral honey prices such as New Zealand or Turkey. In New Zealand, commercial beekeeping has become the next industry that can and should be the target of livestock breeding efforts. The specific biology of A. mellifera (polyandry, sperm storage, mating on the wing) brings with it some obstacles to a structured breeding program through the absence of a traditional pedigree. As a result, no efforts have been made to introduce a structured breeding program in New Zealand. Currently, neither honey producers nor dedicated queen breeders have a way to verify progress made on a stud level in a commercial operation. By using genomic information, the gap of missing pedigree information can be addressed. In collaboration with a large-scale beekeeping company, we have developed strategies for data collection and genotyping in a commercial population of Apis mellifera Ligustica that is continuously supplied with elite queens. With the help of genomic data, breeders are enabled to establish large-scale progeny tests, manage genetic diversity, and provide producers with genetics ideally suited for their production system. With the experience gained in a small high-value market like New Zealand, we hope to provide a basis for similar developments in other countries in the future.

Birefringence or double refraction is the decomposition of a ray of light into two rays when it passes through an aniso- tropic material such as quartz. Sperm cells have been demonstrated to be optically anisotropic. The objective of this study was to evaluate the relationship between the pattern of human sperm head birefringence (SHBF) and DNA damage. A total of 26 patients with normal semen were included. DNA damage (fragmentation and denaturation) was evaluated in the sperm head in the context of birefringence, both total (SHBF-T) and partial (SHBF-P), by terminal deoxyribonucleotidyl transferase (TdT)-mediated dUDP nick-end labelling assay and acridine orange fluorescence, respectively. Positive DNA fragmentation in spermatozoa with SHBF-T (205/1053; 19.5%) was significantly higher (P < 0.0001) than in spermatozoa that presented SHBF-P (60/820; 7.3%). However, the percentage of denatured DNA in spermatozoa with SHBF-T (824/1256; 65.6%) was not significantly different from the ones with SHBF-P (666/1009; 66.0%). In conclusion, the data support a positive relationship between spermatozoa with total SHBF in their head and increased DNA fragmentation. RBMOnline

Summary The aim of this investigation was to determine the presence of abnormal sperm chromatin packaging in spermatozoa with large nuclear vacuoles (LNV) selected via high magnification by analysing the pattern of chromomycin A3 (CMA3) staining. A prospective observational study was designed to analyse semen samples obtained from 66 men undergoing infertility diagnosis and treatment. The numbers of cells with normal (dull yellow staining of the sperm head/CMA3-negative) and abnormal (bright yellow fluorescence of the sperm head/CMA3-positive) chromatin packaging were determined on slides with normal and LNV spermatozoa. The presence of bright yellow fluorescence (CMA3-positive) was significantly higher (p

Summary The aim of this study was to determine the extent of DNA fragmentation and the presence of denatured single-strand or normal double-strand DNA in spermatozoa with extruded nuclear chromatin (ENC) selected by high magnification. Fresh semen samples from 55 patients were prepared by discontinuous isolate concentration gradient. Spermatozoa with normal nucleus (NN) and ENC were selected at 8400· magnification and placed on different slides. DNA fragmentation was determined by TUNEL assay. Denatured and doublestranded DNA was identified by the acridine orange fluorescence method. DNA fragmentation was not significantly different (p = 0.86) between spermatozoa with ENC (19.6%) and those with NN (20%). However, the percentage of spermatozoa with detectable denatured-stranded DNA in the ENC spermatozoon group (59.1%) was significantly higher (p < 0.0001) than in the NN group (44.9%). The high level of denatured DNA in spermatozoa with ENC suggests premature decondensation and disaggregation of sperm chromatin fibres. The results show an association between ENC and DNA damage in spermatozoa, and support the routine morphological selection and injection of motile spermatozoa at high-magnification intracytoplasmic sperm injection.

Regression analysis of 538 semen samples demonstrated that percentages of normal nuclear sperm and all spermatozoa with abnormalities of nuclear form at high magnification had significant negative correlation with percentages of DNA fragmentation. On the other hand, there was a positive correlation between percentages of spermatozoa with nuclear vacuoles and those with DNA fragmentation.

The initiation of the immunological processes leading to type I diabetes is still not understood. The potential of endogenous peptides to induce autoimmune reactions was investigated. Peptides generated in the β-cell by proteolysis could bypass antigen processing by binding to MHC molecules. Selfreactive T and B lymphocytes could be activated by these MHC peptide complexes. The antibody production of peptide-induced B lymphocytes was investigated in mice. Insulin A chain, B chain, C-peptide or amylin were tested for potential induction of antibodies to antigens other than the immunizing peptide. Lymph node B lymphocytes were characterized with an avidin at solid phase ELISA-spot assay. In BALB/c mice insulin A chain induced more spots to B29biotin- and to B1biotinDOP insulin than to A chain itself (P < 0.01, each). Spots to insulin were not inhibited by insulin A chain. Spots to B1DOP insulin were not inhibited by A chain or insulin, excluding crossreaction. Inbred strains of mice with H-2 d but not with H-2 k or H-2 b showed the effect. Application of A chain without adjuvant produced the effect. The antigens recognized by A chain-induced B lymphocytes had to be included in the natural IgM antibody repertoire of the spleen. The study supports the hypothesis that endogenous breakdown peptides can bypass antigen processing resulting in an autoreactive T-B cell interaction. A potential to induce type I diabetes could exist.

The present study aimed to evaluate the correlation between the motile sperm organelle morphology examination (MSOME) and a well-known sperm morphology classification (Tygerberg criteria). For MSOME, spermatozoa were analysed at ×8400 magnification by inverted microscope equipped with Nomarski differential interference contrast optics, Uplan Apo ×100 oil/1.35 objective lens and variable zoom lens. By Tygerberg criteria, the semen underwent morphological evaluation as described in the literature. Regression analysis demonstrated significant positive correlation between percentage of normal sperm forms by Tygerberg criteria and by MSOME (r = 0.83, P < 0.0001). However, the incidence of normal spermatozoa by Tygerberg criteria (9.4%) was significantly higher ( P < 0.0001) than under MSOME (3.3%). Despite the highly positive correlation, MSOME is a much stricter criterion of sperm morphology classification, since it identifies vacuoles and chromatin abnormalities that are not evaluated with the same precision by the analysis of Tygerberg criteria. MSOME should be included among the routine criteria for semen analysis. In addition, MSOME should be used for selection of spermatozoa for intracytoplasmic sperm injection based on the already published literature, as this is a good selection tool.

The aim of this study was to determine the extent of DNA fragmentation and the presence of denatured single-stranded or normal double-stranded DNA in spermatozoa with large nuclear vacuoles (LNV) selected by high magnification. Fresh semen samples from 30 patients were prepared by discontinuous isolate concentration gradient. Spermatozoa with normal nucleus (NN) and LNV were selected at x8400 magnification and placed on different slides. DNA fragmentation was determined by TUNEL assay. Denatured and double-stranded DNA was identified by the acridine orange fluorescence method. DNA fragmentation in spermatozoa with LNV (29.1%) was significantly higher (P < 0.001) than in spermatozoa with NN (15.9%). Therefore, cleavage of genomic DNA in low molecular weight DNA fragments (mono- and oligonucleosomes), and single-strand breaks (nicks) in high molecular weight DNA occur more frequently in spermatozoa with LNV. Similarly, the percentage of denatured-stranded DNA in spermatozoa with LNV (67.9%) was significantly higher (P < 0.0001) than in spermatozoa with NN (33.1%). The high level of denatured DNA in spermatozoa with LNV suggests precocious decondensation and disaggregation of sperm chromatin fibres. The results show an association between LNV and DNA damage in spermatozoa, and support the routine morphological selection and injection of motile spermatozoa at high magnification for ICSI.

The scaphitid ammonite genus Ponteixites Warren, 1934, has remained poorly known and poorly represented in collections up to the present. All previously described specimens of Ponteixites , including the original type specimens, are small and appear to be juveniles. A larger, clearly adult specimen was recently discovered in the Pierre Shale of eastern Montana. If not for the larger size, it would be identified as P. robustus Warren, 1934. A comparison of the new, larger specimen with hundreds of typical, smaller specimens of P. robustus provides evidence for replacing the name Ponteixites with Rhaeboceras Meek, 1876. An alternative interpretation of the large size of the new specimen would be gigantism. However, it is within the size range of other known Rhaeboceras species and has normal scaphite adult characteristics, thus disqualifying it as a candidate for gigantism. The species P. robustus should be renamed R. robustus . Another species, R. coloradoense Cobban, 1987, strongly resembles R. robustus and might be the latter species. The species formerly described as P. gracilis Warren, 1934, is believed either to represent the juvenile stage of another Rhaeboceras species or to be within the range of morphologic variation for R. robustus .

Purpose: to compare the efficacy of recombinant LH supplementation for controlled ovarian stimulation in recombinant FSH and GnRH-agonist protocol. Methods: Search strategies included on-line surveys of databases. The fixed effects model was used for odds ratio and effect size (weighted mean difference). Four trials fulfilled the inclusion criteria. Results: a fewer days of stimulation (p

IL-1beta released from immune cells induces beta cell pro-apoptotic signalling via mitogen-activated protein kinases (MAPKs) and nuclear factor-kappaB (NF-kappaB). In neurons, the neural cell adhesion molecule (NCAM) signals to several elements involved in IL-1beta-induced pro-apoptotic signalling in beta cells. Pancreatic beta cells express NCAM, but its biological effects in these cells are unclear. The aim of this study was to investigate whether there is cross-talk between NCAM signalling and cytokine-induced pro-apoptotic signalling.Western blotting was used to investigate levels of NCAM and inducible nitric oxide synthase, phosphorylation of Src and MAPKs, and cleavage of caspase-3. MAPK activity was investigated with an in vitro kinase assay. Apoptosis was detected by cleaved caspase-3 and a Cell Death Detection ELISA(plus) assay. NCAM-induced fibroblast growth factor receptor (FGFR) activation was investigated in NCAM(-/-) Trex293 cells where FGFR phosphorylation was measured by Western blotting after NCAM transfection.Pre-exposure of INS-1E cells to the FGFR-inhibitor SU5402, but not to the Src-inhibitor PP2, dose-dependently inhibited IL-1beta-mediated MAPK activity. A synthetic peptide, C3d, reported to bind NCAM, did not activate MAPK or Akt as reported in neurons but inhibited IL-1beta-induced MAPK activity, thereby mimicking the effect of SU5402. Furthermore, C3d inhibited NCAM-induced FGFR phosphorylation and apoptosis induced by IL-1beta plus IFN-gamma, but did not affect IL-1beta-induced NF-kappaB signalling.We suggest that NCAM signalling through FGFR is required for efficient IL-1beta pro-apoptotic signalling by facilitating IL-1beta-induced MAPK activation downstream of the NF-kappaB-MAPK branching point. Further, these data identify a novel function of C3d as an inhibitor of NCAM-induced FGFR activity and of IL-1beta-induced MAPK activation in beta cells.

Objective: The aim of this study was to evaluate the ef- fects of aging on semen quality in a population of infertile couples. Methods: A cross-sectional study of semen samples ob- tained from 1,500 men randomly selected from couples who attended an infertility clinic was conducted. The anal- yses were performed using Spearman's correlation and Mann-Whitney tests. The age groups consisted of men ≤ 35 years, from 36-45 years and > 45 years of age. The semen analysis was performed according to the WHO criteria, and morphology was evaluated using the motile sperm organelle morphology examination (MSOME). The percentages of normal spermatozoa and spermatozoa with large nuclear vacuoles (LNV, occupying > 50% nuclear area) were determined. The percentages of DNA fragmen- tation were assessed using the TUNEL assay. Results: A regression analysis revealed that the per- centages of LNV spermatozoa and sperm DNA frag- mentation positively correlated with age. Conversely, a regression analysis revealed that the percentage of nor- mal sperm, sperm progressive motility and sperm vi- tality negatively correlated with age. As in the previous test, the analysis by age group showed that there was a significant reduction (P < 0.05) in the percentage of normal sperm, sperm progressive motility and sperm vitality as age increased. Conversely, the percentage of spermatozoa with LNVs and sperm DNA fragmenta- tion significantly increased (P < 0.05) as age increased. Conclusion: Semen quality seems to be influenced by ag- ing. The age-related decrease in sperm quality suggests that delaying childbearing, not only for women but also for men, may jeopardize reproductive capacity.

The objective of the present study was to determine the importance of the site of embryo transfer (upper or lower half endometrial cavity) on implantation and clinical pregnancy rates.A total of 400 transfers guided by ultrasound were randomly assigned to two groups according to the distance between the uterine fundus and the catheter tip at the time of embryo placement. Group I (n = 200) consisted of transfers corresponding to a distance of50% of the endometrial cavity length (ECL), i.e. transfer in upper half of the cavity; and group II (n = 200) consisted of transfers corresponding to a distance ofor = 50%, of the ECL, i.e. transfer in lower half of cavity. The Student's t-test, Mann-Whitney test and Fisher's exact test were used where appropriate.The general characteristics of the study population and the main transfer cycle characteristics had an equal distribution (P0.05) between groups I and II. No significant difference in implantation or pregnancy rates was observed between groups I and II.The implantation or pregnancy rates were similar whether the embryos were deposited in the upper or lower half of the endometrial cavity.

Purpose: The objective of the present study was to evaluate an ultrasonographic uterine scoring system as a method for the prognosis of embryo implantation in patients submitted to ICSI. Methods: A total of 562 patients submitted to an ICSI program were prospectively evaluated on the day of hCG administration in terms of the following ultrasonographic uterine parameters: A. Endometrial thickness ( 3 = 0; 2.2 a 3 = 2; ≤2.19 = 3); E. Endometrial power Doppler (The endometrium was divided into four equal quadrants and classified as grade I = 1; II = 2; III = 3; IV = 4, according to the visualization of the power Doppler in the quadrants). The colour Doppler signal was considered to be positive when it reached at least the basal layer of the endometrium; F. Myometrial power Doppler (absent = 0; weakly present = 2; strongly present = 3); G. Myometrial echogenicity (homogeneous = 2; inhomogeneous = 0). The patients were divided into 4 groups according to total score: Group I, score

Purpose: The objective of this study was to determine if the zona thinning (ZT) technique improved the rates of implantation and clinical pregnancy for patients aged ≥38 years submitted to an ICSI program.

Summary This study aimed to analyse whether the functional quality of spermatozoa is associated with body mass index (BMI). Semen samples were obtained from 1824 men undergoing fertility evaluation/treatment. Semen analysis was performed using World Health Organization (WHO) criteria, and morphology was evaluated with the motile sperm organelle morphology examination (MSOME). The percentages of sperm DNA fragmentation (using TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick-end labelling (TUNEL) assays), sperm chromatin packaging/underprotamination (using chromomycin A3/CMA3), mitochondrial damage (using MitoTracker Green) and apoptosis (using annexin V) were also assessed. At least 200 spermatozoa were examined in each evaluation. The following BMI values were used as cut-off points: ≤24.9 kg/m2, 25–29.9 kg/m2 (overweight) and ≥30 kg/m2 (obese). High BMI negatively affects sperm concentration, vitality, motility and morphology (p .05). However, increased BMI is associated with increased mitochondrial damage in spermatozoa (p

The aim of this study was to evaluate the effects of aging on semen quality in a population of infertile couples.A cross-sectional study of semen samples obtained from 1,500 men randomly selected from couples who attended an infertility clinic was conducted. The analyses were performed using Spearman's correlation and Mann-Whitney tests. The age groups consisted of men ≤ 35 years, from 36-45 years and45 years of age. The semen analysis was performed according to the WHO criteria, and morphology was evaluated using the motile sperm organelle morphology examination (MSOME). The percentages of normal spermatozoa and spermatozoa with large nuclear vacuoles (LNV, occupying50% nuclear area) were determined. The percentages of DNA fragmentation were assessed using the TUNEL assay.A regression analysis revealed that the percentages of LNV spermatozoa and sperm DNA fragmentation positively correlated with age. Conversely, a regression analysis revealed that the percentage of normal sperm, sperm progressive motility and sperm vitality negatively correlated with age. As in the previous test, the analysis by age group showed that there was a significant reduction (P0.05) in the percentage of normal sperm, sperm progressive motility and sperm vitality as age increased. Conversely, the percentage of spermatozoa with LNVs and sperm DNA fragmentation significantly increased (P0.05) as age increased.Semen quality seems to be influenced by aging. The age-related decrease in sperm quality suggests that delaying childbearing, not only for women but also for men, may jeopardize reproductive capacity.

Purpose: The objective of this study was to assess first embryo cleavage (FEC) 25–27 h after intracytoplasmic sperm injection (ICSI) as a parameter for the embryo selection process. Methods: From January 1998 to December 1999, a total of 670 patients were subjected to the ICSI programme at the Centre for Human Reproduction, Sinh´ Junqueira Maternity Foundation, and the FEC parameter was evaluated in three situations. Results: In the first, a total of 300 zygotes were analyzed on the basis of a score (16–18 h after ICSI) and observed for the presence or absence of FEC (25–27 h after ICSI). A significant (p < 0.02) presence of FEC was observed in zygotes with a score of 15 (ideal score). In the second, a total of 200 patients were selected and divided into two groups matched for age and laboratory performance. Group I (n = 100) was subjected to transfer of embryos with the absence of FEC only (since in this cycle no embryos with FEC were detected within 25–27 h after ICSI) and Group II (n = 100) was subjected to transfer of embryos with the presence of FEC only. The age of Group I patients (33.8 ± 4.2 years) did not differ significantly (p = 0.50) from that of Group II patients (33.5 ± 4.3 years). The number of embryos transferred was similar (p = 0.07) for Group I (2.7 ± 1.1) and Group II (2.9 ± 0.88). In Group II, the 17.5% implantation rate was significantly higher (p < 0.01) than the 5.9% rate obtained for Group I. The pregnancy rate for Group II was significantly higher (p < 0.01) (33%) than that for Group I (12%). The incidence of abortion was 16.6% in Group I as compared with 6% in Group II. In the third situation, we observed the frequency of embryos with FEC in 36 patients whose implantation rate was 100% (ideal result) and obtained a value of 82%. Conclusions: The data suggest that the presence of the FEC parameter that was evaluated 25–27 h after ICSI could be used to select embryos with a higher implantation power. The data reported here may justify routine analysis of embryos with FEC for the process of embryo selection after ICSI.

Purpose: A prospective study was conducted to compare the efficiency of ovarian blockade with nafarelin versus leuprolide in a population whose indication for assisted reproduction was the male factor. Methods: A total of 238 patients were assigned at random to two types of treatment: Group I (119 aspirations), nafarelin (Synarel, Searle), 200 μg by the nasal route twice a day, and Group II (119 aspirations), leuprolide acetate (Lupron, Abbott), 0.5 mg by the subcutaneous route once a day. Both types of blockade were started during the 2nd phase (21st day of the menstrual cycle) and were continued until the day of HCG (5,000–10,000 IU). All patients received a fixed dose of recombinant FSH for 7 days and on the 8th day of stimulation the doses started to be adapted according to ovarian response. Results: Patients' age did not differ (p = 0.93) between Group I (34.1 ± 3.79) and Group II (34 ± 4.64). The number of oocytes retrieved from Group I (10.5 ± 5.93) was also similar (p = 0.57) to that retrieved from Group II (10.2 ± 6.36). In addition, there was no difference (p = 0.58) in the number of oocytes retrieved at Metaphase II between Group I (8.2 ± 4.61) and Group II (7.9 ± 5.2). Fertilization rates and embryo scores were similar (p = 0.81 and p = 0.25, respectively) for Group I (67.5% ± 21.3 and 34.4% ± 14.4) and Group II (68.1% ± 23 and 32.2% ± 14.7, respectively). In addition, pregnancy rates per puncture and per embryo transfer and implantation rates were similar for Group I (30.2, 31.3, and 16.2%, respectively) then compared with Group II (24.4, 25.2, and 12.6%, respectively) (p = 0.38, p = 0.37, and p = 0.22, respectively). Conclusions: Implantation and pregnancy rates per puncture and per embryo transfer are not statistically significant in the nafarelin group when compared with leuprolide group.

Purpose:The aim of this prospective, randomized study was to compare the results obtained in ICSI with two culture media, P-1 (Irvine Scientific) and IVF-50 (Scandinavian IVF Science). Methods: A total of 182 patients undergoing ICSI treatment were randomly included in this study and divided in two groups: Group I: P-1 medium (n = 91) or Group II: IVF-50 medium (n = 91). All the embryos were transferred on the second day. Results: Patient age did not differ (p = .29) between Group I (34.8 ± 4.8) and Group II (34.0 ± 4.5). The number of oocytes retrieved from Group I (10.6 ± 6.7) was also similar (p = .49) to that retrieved from Group II (11.1 ± 6.4). In addition, there was no difference (p = .25) in the number of oocytes retrieved at metaphase II between Group I (7.9 ± 4.6) and Group II (8.7 ± 4.6). Normal fertilization rates, abnormal fertilization rates, and cleavage rates were similar (p = .62, p = .48, and p = .9, respectively) between Group I (68.4 ± 23.3%, 6.7 ± 10.3%, and 98.7 ±4.6%) and Group II (65.3 ± 26.2%, 9.0 ± 13.8%, and 98.9 ± 3.9%, respectively). The embryo score was also similar (p = .62) for both groups (Group I: 31.9 ± 14.0 and Group II: 33.4 ± 15.8). There was no difference in the number of embryos transferred (p = .69) between Group I (2.8 ± 1.0) and Group II (2.8 ± 1.1). In addition, pregnancy rates/puncture, pregnancy rates/transfer, implantation rates, and abortion rates were also similar for Group I (36.2%, 37.0%, 17.4%, and 12.1%, respectively) and Group II (31.8%, 33.7%, 15.8%, and 10.3%, respectively) (p = .64, p = .75, p = .72, and p = 1.0, respectively). Conclusions: There were no differences in the results obtained with culture media P-1 (Irvine Scientific) and IVF-50 (Scandinavian IVF Science) for ICSI and embryo culture.

Purpose: Our purpose was to compare an ultrarapid method (URM) modified with dimethyl sulfoxide (Me2SO) to a slow method (SM) with propanediol (PROH) for the cryopreservation of extra human embryos in a program of intracytoplasmic sperm injection (ICSI). Methods: The extra embryos of 160 patients were cryopreserved in a prospective and randomized manner (drawing lots) by a modified URM (3 M Me2SO/0.25 M sucrose/thawing in three sucrose gradients) (Group I) or by a SM )1.5 M Propanediol/program 0-Cryologic CL863) (Group II). A total of 103 cycles has been thawed thus far. The number of thawed cycles was 58 for group I and 45 for group II. Results: The mean age (group I, 31.3 ± 4.5; group II, 31.9 ± 4.3) did not differ between the groups (P = 0.38). The number of frozen embryos (group I, 6.6 ± 3.2; group II, 6.5 ± 3.2) was similar (P = 0.49) for the two groups, as was the number of thawed embryos (P = 0.52) (group I, 6.5 ± 2.9; group II, 6.2 ± 3). The survival rate was higher (P < 0.01) for group II (83.3 ± 23%) than for group I (69.2 ± 28.7%). The cleavage rate was also higher (P < 0.01) for group II (56.8 ± 31%) compared with group I (24.2 ± 22.4%). The number of embryos transferred did not differ (P = 0.14) between the groups (group I, 3.16 ± 1.2; group II, 3.5 ± 1.0). The implantation rate (group I, 6.3%; group II, 13.8%) was significantly different between groups (P = 0.034). Pregnancy rates per thawed and transferred cycle were higher for group II (33.3 and 36.6%, respectively) compared with group I (13.8 and 16%, respectively), and these differences were significant (P = 0.03 and P = 0.03, respectively). Conclusion: The data obtained suggest that the SM is superior to the URM for the cryopreservation of extra embryos after ICSI.

Purpose: Zona thinning (ZT) is a technique used to improve pregnancy rates among patients ≥38 years old and/or patients presenting previous implantation failure. The objective of the study was to determine whether ZT has a beneficial effect on patients younger than 37 years who are undergoing the first ICSI attempt.

Non-haematopoietic hepatic malignancies are uncommon in birds. The clinical presentation (i.e. chronic buphthalmos)and non-specific radiographic findings observed in this adult Amazon parrot (Amazona spp.) were not consistent with previous reports describing the natural behaviour of metastatic hepatocellular carcinoma in birds.

The expression of self-incompatibility (SI) rejection patterns in 29 accessions of wildBeta species was studied at the anatomical level. Three different SI-related pollen tube rejection patterns were observed: (1) The pollen germinate from many pores and the pollen tubes are very short, often with swollen tips. (2) The pollen germinate and grow along the stigmatic papillae, where the self-incompatibility response takes place at the basis of the stigmatic papillae. (3) The pollen germinate and the pollen tubes grow with a distinct direction towards the style; the self-incompatibility response is observed at the border between the stigma and the style. In general the patterns observed inB. vulgaris L. subsp.vulgaris sensu lato seem to be in accordance with an ongoing microevolutionary process that has given rise to the largemaritima complex (viz.B. trojana Pamukcuoglu,B. atriplicifolia Rouy, andB. maritima (L.) Arcangeli), in which it seems that the variation is related to geographical origin. The data support the idea that the endemicB. vulgaris subsp.adanensis (Pamukcuoglu) Ford-Lloyd & Williams forms a distinct, self-compatible group separated from themaritima complex and also thatB. macrocarpa Gussone should be retained as a separate self-compatible species. For the sectionCorollinae Ulbrich the rejection patterns are in accordance with a division into two subsections (viz.,Lomatogonae andTrigynae) previously suggested on the basis of phenotypical and molecular data. The results fromProcumbentes Ulbrich are interpreted as evidence forB. procumbens Smith andB. webbiana Moquin being more related to each other than toB. patellaris Moquin, reflecting also a similar taxonomic relationship.

The objective of this work was to evaluate the results obtained with a protocol of semi-programmed ovarian stimulation (low-dose contraceptive pill+clomiphene citrate+human menopausal gonadotrophin+dexamethasone) used as the first-choice method for in-vitro fertilization (IVF). A total of 207 punctures was performed for oocyte collection from 168 patients (mean age 31.0±4.0 years); mean infertility duration was 5.81±3.30 years. The infertility factors indicating IVF for this population were as follows: tubo-peritoneal factor, 68%; pure or associated male factor, 9.2%; endometriosis, 11.1%; ovulatory factor, 4.3%; idiopathic factor, 11.6%; others, 2.4%. No oocyte was found on aspiration in five procedures (2.4%), with the mean number of oocytes collected per cycle being 5.87±3.3 (range 0-18). The cancellation rate per puncture was 5%. The mean embryo cleavage rate was 60.2±36.8%, with transfer of at least one embryo occurring in 82.6% of all punctures. The mean number of transferred embryos was 2.52±1.60 (range 1-5). The clinical pregnancy rates per started cycle and per puncture were 22.4 (218 ovarian stimulation cycles) and 23.6% (a total of 49 clinical pregnancies, 36 single, nine twins and four triplets) respectively. The clinical pregnancy rate per embryo transfer was 28.6%. The embryo implantation rate was 12.6%. The abortion rate was 16.3%. The index of deliveries per puncture was 19.8%. There were no cases of moderate or severe ovarian hyperstimulation syndrome. The favourable results obtained, in addition to the low operational costs, confirm the validity of the use of semiprogrammed cycles as the first choice for patients undergoing the IVF process

Made available in DSpace on 2013-08-28T14:07:11Z (GMT). No. of bitstreams: 1 WOS000314711300001.pdf: 1191127 bytes, checksum: d0eabe31dd3830314003d7c40434c14a (MD5) Made available in DSpace on 2013-09-30T18:20:43Z (GMT). No. of bitstreams: 2 WOS000314711300001.pdf: 1191127 bytes, checksum: d0eabe31dd3830314003d7c40434c14a (MD5) WOS000314711300001.pdf.txt: 50115 bytes, checksum: 7302ae309486d05178dc76dbe7d3a0d6 (MD5) Previous issue date: 2012-11-21 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:35:40Z No. of bitstreams: 2 WOS000314711300001.pdf: 1191127 bytes, checksum: d0eabe31dd3830314003d7c40434c14a (MD5) WOS000314711300001.pdf.txt: 50115 bytes, checksum: 7302ae309486d05178dc76dbe7d3a0d6 (MD5) Made available in DSpace on 2014-05-20T13:35:40Z (GMT). No. of bitstreams: 2 WOS000314711300001.pdf: 1191127 bytes, checksum: d0eabe31dd3830314003d7c40434c14a (MD5) WOS000314711300001.pdf.txt: 50115 bytes, checksum: 7302ae309486d05178dc76dbe7d3a0d6 (MD5) Previous issue date: 2012-11-21 Background: The objective was to present a new ovarian response prediction index (ORPI), which was based on anti-Mullerian hormone (AMH) levels, antral follicle count (AFC) and age, and to verify whether it could be a reliable predictor of the ovarian stimulation response.Methods: A total of 101 patients enrolled in the ICSI programme were included. The ORPI values were calculated by multiplying the AMH level (ng/ml) by the number of antral follicles (2-9 mm), and the result was divided by the age (years) of the patient (ORPI=(AMH x AFC)/Patient age).Results: The regression analysis demonstrated significant (P

Made available in DSpace on 2013-08-28T14:08:29Z (GMT). No. of bitstreams: 1 WOS000308910500001.pdf: 221658 bytes, checksum: d864f7f93f4a2cdcf5f16330cfc3ef0a (MD5) Made available in DSpace on 2013-09-30T18:20:42Z (GMT). No. of bitstreams: 2 WOS000308910500001.pdf: 221658 bytes, checksum: d864f7f93f4a2cdcf5f16330cfc3ef0a (MD5) WOS000308910500001.pdf.txt: 51849 bytes, checksum: feaa0df4b446b5e195b5e9d457f76ca4 (MD5) Previous issue date: 2012-06-15 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:35:39Z No. of bitstreams: 2 WOS000308910500001.pdf: 221658 bytes, checksum: d864f7f93f4a2cdcf5f16330cfc3ef0a (MD5) WOS000308910500001.pdf.txt: 51849 bytes, checksum: feaa0df4b446b5e195b5e9d457f76ca4 (MD5) Made available in DSpace on 2014-05-20T13:35:39Z (GMT). No. of bitstreams: 2 WOS000308910500001.pdf: 221658 bytes, checksum: d864f7f93f4a2cdcf5f16330cfc3ef0a (MD5) WOS000308910500001.pdf.txt: 51849 bytes, checksum: feaa0df4b446b5e195b5e9d457f76ca4 (MD5) Previous issue date: 2012-06-15 Background: It is not well established whether the increased number of leukocytes in the seminal fluid impairs the outcomes of assisted reproductive technology (ART). This investigation analysed the outcomes of the intracytoplasmic sperm injection (ICSI) and intracytoplasmic morphologically selected sperm injection (IMSI) cycles in couples in which the male partner exhibited leukocytospermia.Methods: A total of 100 cycles in 100 couples were included in this study. For the ICSI or IMSI procedures, the patients were divided into two groups according to the presence or absence of leukocytospermia and then matched by (female) age:- ICSI: Group I (n = 25): Leukocytospermia - semen samples with a leukocyte count of greater than or equal to 1 x 10(6)/mL; and Group II (n = 25): Non-leukocytospermia - semen samples with a leukocyte count < 1 x 10(6)/mL.- IMSI: Group I (n = 25): Leukocytospermia; and Group II (n = 25): Non-leukocytospermia.The endpoints included the rates of fertilisation, implantation, clinical pregnancy, miscarriage, ongoing pregnancy and live birth. Student's t-tests, Mann-Whitney tests and Chi-square tests were performed, and P < 0.05 was considered significant.Results: The data from the ICSI groups showed that leukocytospermia did not have a negative influence on the rates of fertilisation (Group I: 57.9+/-30.2%, Group II: 61.9+/-27.7%; P = 0.74), implantation (Group I: 12.3%; Group II: 13.5%; P = 0.93), clinical pregnancy (Group I: 24%; Group II: 24%; P = 1.0), miscarriage ( Group I: 0, Group II: 0), ongoing pregnancy (Group I: 24%; Group II: 24%; P = 1.0), or live births (Group I: 24%; Group II: 24%; P = 1.0). Similarly, the data from the IMSI groups also showed that the leukocytospermia did not have a negative influence on the rates of fertilisation (Group I: 67.6+/-24.6%, Group II: 59.5+/-28.1%; P = 0.36), implantation (Group I: 17.5%; Group II: 16.7%; P = 0.90), clinical pregnancy (Group I: 28%; Group II: 24%; P = 1.0), miscarriage (Group I: 14.3%; Group II: 0; P = 0.33), ongoing pregnancy (Group I: 24%; Group II: 24%; P = 1.0), or live births (Group I: 24%, 6/25; Group II: 24%, 6/25; P = 1.0).Conclusions: The results indicate that the leukocytospermia may not have a negative effect on the outcomes of ICSI or IMSI cycles. Nevertheless, it seems that it is necessary to more precisely determine the effects, if any, of seminal leukocytes on fertilisation and implantation processes. Such efforts will help to establish a more reliable leukocyte threshold, which could eventually demonstrate whether there is a negative influence on the ART procedures. Ctr Human Reprod Prof Franco Jr, Ribeirao Preto, Brazil Hosp Perola Byington, Womens Hlth Reference Ctr, São Paulo, Brazil Paulista Ctr Diag Res & Training, Ribeirao Preto, Brazil São Paulo State Univ UNESP, Botucatu Med Sch, Dept Gynecol & Obstet, Botucatu, SP, Brazil São Paulo State Univ UNESP, Botucatu Med Sch, Dept Gynecol & Obstet, Botucatu, SP, Brazil

Made available in DSpace on 2013-08-28T14:09:24Z (GMT). No. of bitstreams: 1 WOS000307307000001.pdf: 639767 bytes, checksum: b78f9de52568816f019ce2c064623acd (MD5) Made available in DSpace on 2013-09-30T18:20:41Z (GMT). No. of bitstreams: 2 WOS000307307000001.pdf: 639767 bytes, checksum: b78f9de52568816f019ce2c064623acd (MD5) WOS000307307000001.pdf.txt: 49302 bytes, checksum: 51e5f2f668acfe2b84c709224143ca36 (MD5) Previous issue date: 2012-04-27 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:35:39Z No. of bitstreams: 2 WOS000307307000001.pdf: 639767 bytes, checksum: b78f9de52568816f019ce2c064623acd (MD5) WOS000307307000001.pdf.txt: 49302 bytes, checksum: 51e5f2f668acfe2b84c709224143ca36 (MD5) Made available in DSpace on 2014-05-20T13:35:39Z (GMT). No. of bitstreams: 2 WOS000307307000001.pdf: 639767 bytes, checksum: b78f9de52568816f019ce2c064623acd (MD5) WOS000307307000001.pdf.txt: 49302 bytes, checksum: 51e5f2f668acfe2b84c709224143ca36 (MD5) Previous issue date: 2012-04-27 Background: The selection of developmentally competent human gametes may increase the efficiency of assisted reproduction. Spermatozoa and oocytes are usually assessed according to morphological criteria. Oocyte morphology can be affected by the age, genetic characteristics, and factors related to controlled ovarian stimulation. However, there is a lack of evidence in the literature concerning the effect of gonadotropin-releasing hormone (GnRH) analogues, either agonists or antagonists, on oocyte morphology. The aim of this randomized study was to investigate whether the prevalence of oocyte dysmorphism is influenced by the type of pituitary suppression used in ovarian stimulation.Methods: A total of 64 patients in the first intracytoplasmic sperm injection (ICSI) cycle were prospectively randomized to receive treatment with either a GnRH agonist with a long-term protocol (n: 32) or a GnRH antagonist with a multi-dose protocol (n: 32). Before being subjected to ICSI, the oocytes at metaphase II from both groups were morphologically analyzed under an inverted light microscope at 400x magnification. The oocytes were classified as follows: normal or with cytoplasmic dysmorphism, extracytoplasmic dysmorphism, or both. The number of dysmorphic oocytes per total number of oocytes was analyzed.Results: Out of a total of 681 oocytes, 189 (27.8 %) were morphologically normal, 220 (32.3 %) showed cytoplasmic dysmorphism, 124 (18.2%) showed extracytoplasmic alterations, and 148 (21.7%) exhibited both types of dysmorphism. No significant difference in oocyte dysmorphism was observed between the agonist- and antagonist- treated groups (P > 0.05). Analysis for each dysmorphism revealed that the most common conditions were alterations in polar body shape (31.3%) and the presence of diffuse cytoplasmic granulations (22.8%), refractile bodies (18.5%) and central cytoplasmic granulations (13.6%). There was no significant difference among individual oocyte dysmorphisms in the agonist- and antagonist-treated groups (P > 0.05).Conclusions: Our randomized data indicate that in terms of the quality of oocyte morphology, there is no difference between the antagonist multi-dose protocol and the long-term agonist protocol. If a GnRH analogue used for pituitary suppression in IVF cycles influences the prevalence of oocyte dysmorphisms, there does not appear to be a difference between the use of an agonist as opposed to an antagonist. São Paulo State Univ UNESP, Dept Gynecol & Obstet, Botucatu Med Sch, Botucatu, SP, Brazil Ctr Human Reprod Prof Franco Jr, Ribeirao Preto, Brazil Paulista Ctr Diag Res & Training, Ribeirao Preto, Brazil Perola Byington Hosp, Womens Hlth Reference Ctr, São Paulo, Brazil São Paulo State Univ UNESP, Dept Gynecol & Obstet, Botucatu Med Sch, Botucatu, SP, Brazil

Johnson KB, Petersen-Jones H, Thompson JM, Hitomi K, Itoh M, Bakker ENTP, Johnson GV, Colak G, Watts SW. Vena cava and aortic smooth muscle cells express transglutaminases 1 and 4 in addition to transglutaminase 2. Am J Physiol Heart Circ Physiol 302: H1355-H1366, 2012. First published February 3, 2012; doi: 10.1152/ajpheart.00918.2011.-Transglutaminase (TG) function facilitates several vascular processes and diseases. Although many of these TG-dependent vascular processes have been ascribed to the function of TG2, TG2 knockout mice have a mild vascular phenotype. We hypothesized that TGs besides TG2 exist and function in the vasculature. Biotin-pentylamide incorporation, a measure of general TG activity, was similar in wild-type and TG2 knockout mouse aortae, and the general TG inhibitor cystamine reduced biotin-pentylamine incorporation to a greater extent than the TG2-specific inhibitor Z-DON, indicating the presence of other functional TGs. Additionally, 5-hydroxytryptamine-induced aortic contraction, a TG-activity-dependent process, was decreased to a greater extent by general TG inhibitors vs. Z-DON (maximum contraction: cystamine = abolished, monodansylcadaverine = 28.6 +/- 14.9%, and Z-DON = 60.2 +/- 15.2% vehicle), providing evidence for the importance of TG2-independent activity in the vasculature. TG1, TG2, TG4, and Factor XIII (FXIII) mRNA in rat aortae and vena cavae was detected by RT-PCR. Western analysis detected TG1 and TG4, but not FXIII, in rat aortae and vena cavae and in TG2 knockout and wild-type mouse aortae. Immunostaining confirmed the presence of TG1, TG2, and TG4 in rat aortae and vena cavae, notably in smooth muscle cells; FXIII was absent. K5 and T26, FITC-labeled peptide substrates specific for active TG1 and TG2, respectively, were incorporated into rat aortae and vena cavae and wild-type, but not TG2 knockout, mouse aortae. These studies demonstrate that TG2-independent TG activity exists in the vasculature and that TG1 and TG4 are expressed in vascular tissues

A solid-phase immunoenzymatic technique with B1- or B29-biotinylated insulin coupled to avidincoated wells was used to characterize serum anti-insulin antibodies and to locate insulin antibody-producing B lymphocytes in different organs of mice. Low natural serum anti-insulin IgM and IgG antibodies were found in ten different healthy inbred strains of mice. Prediabetic nonobese diabetic (NOD) mice had significantly higher measurements than BALB/c mice (P

The objective of the present study was to compare the rate of embryo cleavage after oocyte insemination with capacitated spermatozoa in the presence or absence of follicular fluid. In the first experiment, a total of 67 oocytes were inseminated with spermatozoa which had been submitted to part of the capacitation procedures in the presence of follicular fluid (21 oocytes) and with spermatozoa capacitated only with Menezo B2 culture medium (46 oocytes). Thirteen and 29 cleaved embryos were obtained, respectively (p = 0.89). In the second experiment, a total of 61 oocytes were inseminated with spermatozoa submitted to the entire capacitation procedure in follicular fluid (17 oocytes) and capacitated only with Menezo B2 culture medium (44 oocytes). Eleven and 31 cleaved embryos were obtained, respectively (p = 0.67). The absence of significant differences in cleavage rate between the two experiments indicates that the quality of the spermatozoa capacitation process is not changed by the addition of follicular fluid.