The number of nuclei in compacted embryos, assessed by optical coherence microscopy, is a non-invasive and robust marker of mouse embryo quality.

Optical coherence microscopy (OCM) visualizes nuclei in live, unlabeled cells. As most cells are uninucleated, the number of nuclei in embryos may serve as a proxy of the cell number, providing important information on developmental status of the embryo. Importantly, no other non-invasive method currently allows for the cell number count in compacted embryos. We addressed the question of whether OCM, by providing the number of nuclei in compacted mouse embryos, may help evaluate embryo quality. We subjected compacted embryonic Day 3 (E3.0: 72 h after onset of insemination) mouse embryos to OCM scanning and correlated nuclei number and developmental potential. Implantation was assessed using an outgrowth assay (in vitro model meant to reflect embryonic ability to implant in vivo). Embryos with more cells at E3.0 (>18 cells) were more likely to reach the blastocyst stage by E4.0 and E5.0 (P ≪ 0.001) and initiate hatching by E5.0 (P < 0.05) than those with fewer cells (<12 cells). Moreover, the number of cells at E3.0 strongly correlated with the total number of cells in E4.0 and E5.0 embryos (ρ = 0.71, P ≪ 0.001 and ρ = 0.61, P ≪ 0.001, respectively), also when only E4.0 and E5.0 blastocysts were considered (ρ = 0.58, P ≪ 0.001 and ρ = 0.56, P ≪ 0.001, respectively). Additionally, we observed a strong correlation between the number of cells at E3.0 and the number of trophectoderm cells in E4.0 and E5.0 blastocysts (ρ = 0.59, P ≪ 0.001 and ρ = 0.57, P ≪ 0.001, respectively). Importantly, embryos that had more cells at E3.0 (>18 cells) were also more likely to implant in vitro than their counterparts with fewer cells (<12 cells; P ≪ 0.001). Finally, we tested the safety of OCM imaging, demonstrating that OCM scanning affected neither the amount of reactive oxygen species nor mitochondrial activity in the embryos. OCM also did not hinder their preimplantation development, ability to implant in vitro, or to develop to term after transfer to recipient females. Our data indicate that OCM imaging provides important information on embryo quality. As the method seems to be safe for embryos, it could be a valuable addition to the current repertoire of embryo evaluation methods. However, our study was conducted only on mouse embryos, so the proposed protocol would require optimization in order to be applied in other species.

Spindle shape and volume differ in high- and low-quality metaphase II oocytes.

In brief: Optical coherence microscopy non-invasively visualizes metaphase II spindles allowing for quantitative analysis of their volume and shape, which may prove useful in the assessment of the oocyte quality. Using a mouse model, we showed also that analysis of spindle length combined with morphokinetics improves the evaluation of the resulting embryos. Abstract: The proper development of embryos strongly depends on the quality of oocytes, so the evaluation of oocytes may be a useful initial step in IVF procedures. Additionally, it enables embryologists to make more informed decisions regarding the treatments chosen for the patients and better manage patients' expectations. Optical coherence microscopy (OCM) allows for non-invasive 3D visualization of intracellular structures, such as spindles or nuclei, which have been linked to the success of embryonic development. Here, we applied a mouse model to examine whether OCM imaging could be used in the quality assessment of metaphase II (MII) oocytes. We showed that quantitative parameters describing the shape and volume of the MII spindle were associated with the quality of the resulting embryos, including the likelihood of blastocyst formation and the embryos' ability to differentiate the trophectoderm and primitive endoderm, but not the epiblast. We also created a multivariate linear regression model, combining OCM-based quantification of MII spindles with morphokinetic analysis of the embryos, that allowed for improved evaluation of the embryo quality. Finally, we proved that OCM does not interfere with the viability of the scanned cells, at least during the preimplantation development. Therefore, we believe that OCM-based quantitative assessment of MII spindles can improve the oocyte and embryo selection in IVF procedures.

Neutrophil-to-Platelet Ratio in Patients with Ulcerative Colitis Treated with Infliximab or Vedolizumab: A Retrospective, Single-Center Study in Poland.

BACKGROUND This retrospective study from a single center in Poland was undertaken to evaluate the clinical utility of neutrophil-to-platelet ratio in therapy of 35 ulcerative colitis (UC) patients with infliximab or vedolizumab. MATERIAL AND METHODS This study included 35 patients: 16 were treated with infliximab and 19 were treated with vedolizumab. Treatment response was evaluated using partial Mayo score. Treatment response was defined as a reduction of partial Mayo score of ≥3 points followed by a decrease of a minimum of 30% from the baseline, decrease in the rectal bleeding subscore of ≥1, or an absolute rectal bleeding subscore of 0 or 1. During the maintenance period, we diagnosed 13 patients with loss of response (LOR) (5 with infliximab and 8 with vedolizumab). The Mann-Whitney U test was performed to assess differences between the groups. Statistical significance was defined as P<0.05. The median was used to describe the value of the parameter. Analysis of the receiver operating characteristic (ROC) curve with the determination of area under the curve (AUC) was performed for the neutrophil-to-platelet parameter during the induction period. RESULTS The median value of the neutrophil-to-platelet ratio for the treatment response group was lower than in the LOR group (median=13.18 and median=19.49, respectively). Calculation of AUC curve for neutrophil-to-platelet ratio during the induction period showed best sensitivity and specificity for values ≥32.511. There were no other significant findings. CONCLUSIONS Neutrophil-to-platelet ratio might be a promising biomarker of LOR in biologic therapy of UC. However, to fully prove this, further studies are needed.

Flavonoids as tyrosinase inhibitors in in silico and in vitro models: basic framework of SAR using a statistical modelling approach.

Flavonoids are widely distributed in plants and constitute the most common polyphenolic phytoconstituents in the human diet. In this study, the in vitro inhibitory activity of 44 different flavonoids (1-44) against mushroom tyrosinase was studied, and an in silico study and type of inhibition for the most active compounds were evaluated too. Tyrosinase inhibitors block melanogenesis and take part in melanin production or distribution leading to pigmentation diseases. The in vitro study showed that quercetin was a competitive inhibitor (IC50=44.38 ± 0.13 µM) and achieved higher antityrosinase activity than the control inhibitor kojic acid. The in silico results highlight the importance of the flavonoid core with a hydroxyl at C7 as a strong contributor of interference with tyrosinase activity. According to the developed statistical model, the activity of molecules depends on hydroxylation at C3 and methylation at C8, C7, and C3 in the benzo-γ-pyrane ring of the flavonoids.

Effect of systemic antibiotics on the outcomes of regenerative periodontal surgery in intrabony defects: a randomized, controlled, clinical study.

OBJECTIVES: To assess the potential influence of systemic antibiotic administration on the healing of periodontal intrabony defects treated with deproteinized bovine bone mineral (DBBM) and collagen membrane. MATERIALS AND METHODS: Forty-one intrabony defects were treated by means of DBBM and collagen membrane (GTR). Postoperatively, the patients received either systemic antibiotics (i.e., 1 g of amoxicillin, twice daily for 7 days) (test) or no antibiotics (control). Clinical attachment level (CAL), probing depth (PD), and gingival recession (GR) were measured at baseline and at 1 year following regenerative surgery. The depth of the intrabony component (INTRA DD) and its width (INTRA DW) were measured during surgery and after 1 year at reentry. The depth (RxD) and width (RxW) of the intrabony defects were evaluated radiographically at baseline and at 1 year. RESULTS: No adverse events were observed in any of the two groups throughout the entire study period. In the test group, mean CAL changed from 8.7 ± 1.4 mm at baseline to 5.0 ± 1.7 mm at 1 year (p < 0.0001), while PD decreased from 7.8 ± 1.5 mm at baseline to 4.0 ± 0.9 mm at 1 year (p < 0.0001). In the control group, mean CAL changed from 8.6 ± 1.9 mm to 5.9 ± 1.6 mm (p < 0.001) and mean PD improved from 7.4 ± 1.3 mm to 4.1 ± 1.3 mm (p < 0.001). Mean CAL gain measured 3.6 ± 1.6 mm in the test and 2.7 ± 1.6 mm in the control group, respectively. Defect fill (i.e., INTRA DD gain) at re-entry measured 3.7 ± 1.8 mm in the test and 2.7 ± 2.1 mm in the control group. A CAL gain of ≥ 3 mm was measured in 76% of the defects in the test group and in 40% of the defects in the control group, respectively. In both groups, all evaluated clinical and radiographic parameters improved statistically significantly compared with baseline, but no statistically significant differences were found between the two groups. CONCLUSIONS: Within their limits, the present study has failed to show any substantial added clinical benefits following the postoperative administration of amoxicillin in conjunction with regenerative periodontal surgery using DBBM and GTR. CLINICAL RELEVANCE: The post-surgically administration of systemic antibiotics does not seem to be necessary following regenerative periodontal surgery.

Clinical evaluation of Miller class I and II recessions treatment with the use of modified coronally advanced tunnel technique with either collagen matrix or subepithelial connective tissue graft: A randomized clinical study.

AIM: To compare outcomes of modified coronally advanced tunnel technique (MCAT) combined with either collagen matrix (CM) or subepithelial connective tissue graft (SCTG) in the treatment of Miller class I and II multiple gingival recessions in the mandible. MATERIALS AND METHODS: The study encompassed 91 recessions in 29 patients for whom MCAT was combined with CM on one side of the mandible and SCTG on the contralateral one. The following clinical parameters were measured: gingival recession height (GR) and width (RW), probing depth (PD), clinical attachment level (CAL), width of keratinized tissue (KT), gingival thickness (GT), mean (MRC) and complete root coverage (CRC) and Root Coverage Esthetic Score (RES). RESULTS: The MRC proportions on the CM- and SCTG-treated sides were 53.20% and 83.10%, respectively. CRC was achieved in 9 out of 45 (20%) gingival defects treated with CM and 31 out of 46 (67%) treated with SCTG. There were statistically significant differences in MRC, CRC, GR, RW, KT, GT and RES between CM- and CTG-treated sides. CONCLUSIONS: Modified coronally advanced tunnel technique leads to reduction in gingival recession both when combined CM and SCTG, of which the latter is more efficient as far as root coverage and aesthetic parameters are concerned.

Plasma concentration of MMP-1 and MMP-2 in boys with cryptorchidism and its lack of correlation with INSL3 and inhibin B.

The matrix metalloproteinases are enzymes capable of remodeling of extracellular matrix, and modulate the behavior of cells. Maturation of gubernaculum and spermatogenesis demand proper equilibrium of metalloproteinases and their inhibitors. The aim of this survey was to investigate the levels of matrix metalloproteinase type 1 (MMP-1) and matrix metalloproteinase type 2 (MMP-2) in the plasma of children with unilateral cryptorchidism along with levels of Insulin-like Peptide 3 (INSL3) and inhibin B. INSL3 have a role in gubernaculum development. Inhibin B is produced by Sertoli cells, and its levels reflect the status of the testis germinative epithelium. Fifty boys with an undescended testicle, aged 1-4 years (median = 2.4 years) were enrolled into the study. Fifty boys with inguinal hernia aged 1-4 years, served as controls (median age = 2.1 years). Investigators assessed the MMP-1 and MMP-2 concentrations using Surface Plasmon Resonance Imaging. The levels of INSL-3 and inhibin B were assessed using commercial enzyme-linked immunosorbent assay ELISA. The median concentration of MMP-1 and MMP-2 in the blood plasma of patients with unilateral cryptorchidism, was nearly 2-folds higher than in controls. The great area under the ROC curve with the cut off value of 0.865 for MMP-1, and 0.819 for MMP-2, indicates the high clinical sensitivity and specificity of the test of plasma levels of MMP-1 and MMP-2 for boys with cryptorchidism. The increased plasma levels of MMP-1 and MMP-2, probably reflect the level of apoptosis of the germ cells in undescended testicles, in response to the heat stress during the period of prepubertal testis development. In the group of cryptorchid boys, we found slightly lower concentrations of INSL3, without statistical significance and without correlation with MMP-1 and MMP-2 levels. There were no significant differences in the levels of inhibin B in the group of boys with cryptorchidism and boys with inguinal hernia and it also did not correlate with MMP-1 and MMP-2 concentrations.

Simultaneous analysis of bisphenol A fractions in maternal and fetal compartments in early second trimester of pregnancy.

Background Bisphenol A (BPA) is an estrogenic, endocrine-disrupting compound widely used in the industry. It is also a ubiquitous environmental pollutant. Its presence was confirmed in human fetuses, which results from maternal exposure during pregnancy. The mechanisms behind maternal-fetal transfer, and relationships between pregnant women and fetal exposures remain unclear. The aim of this study was to assess the impact of maternal exposure to BPA on the exposure of the fetus. Methods Maternal plasma and amniotic fluid samples were collected from 52 pregnant women undergoing amniocentesis for prenatal diagnosis of chromosomal abnormalities. BPA was measured by gas chromatography-mass spectrometry (GC-MS). The permeability factor - a ratio of fetal-to-maternal BPA concentration - was used as a measure delineating the transplacental transfer of BPA. Results The median concentration of maternal plasma BPA was 8 times higher than the total BPA concentration in the amniotic fluid (8.69 ng/mL, range: 4.3 ng/mL-55.3 ng/mL vs. median 1.03 ng/mL, range: 0.3 ng/mL-10.1 ng/mL). There was no direct relationship between the levels of BPA in maternal plasma and amniotic fluid levels. The permeability factor, in turn, negatively correlated with fetal development (birth weight) (R = -0.54, P < 0.001). Conclusion Our results suggest that the risk of fetal BPA exposure depends on placental BPA permeability rather than the levels of maternal BPA plasma concentration and support general recommendations to become aware and avoid BPA-containing products.

Dynamics of cytoplasm and cleavage divisions correlates with preimplantation embryo development.

In vitro fertilization has become increasingly popular as an infertility treatment. In order to improve efficiency of this procedure, there is a strong need for a refinement of existing embryo assessment methods and development of novel, robust and non-invasive selection protocols. Studies conducted on animal models can be extremely helpful here, as they allow for more extensive research on the potential biomarkers of embryo quality. In the present paper, we subjected mouse embryos to non-invasive time-lapse imaging and combined the Particle Image Velocimetry analysis of cytoplasmic dynamics in freshly fertilized oocytes with the morphokinetic analysis of recordings covering 5 days of preimplantation development. Our results indicate that parameters describing cytoplasmic dynamics and cleavage divisions independently correspond to mouse embryo's capacity to form a high-quality blastocyst. We also showed for the first time that these parameters are associated with the percentage of abnormal embryonic cells with fragmented nuclei and with embryo's ability to form primitive endoderm, one of the cell lineages differentiated during preimplantation development. Finally, we present a model that links selected cytoplasmic and morphokinetic parameters reflecting frequency of fertilization-induced Ca2+-oscillations and timing of 4-cell stage and compaction with viability of the embryo assessed as the total number of cells at the end of its preimplantation development. Our results indicate that a combined analysis of cytoplasmic dynamics and morphokinetics may facilitate the assessment of embryo's ability to form high-quality blastocysts.

Assessment of the effect of the corticotomy-assisted orthodontic treatment on the maxillary periodontal tissue in patients with malocclusions with transverse maxillary deficiency: a case series.

BACKGROUND: The aim of the study was to assess the effect of corticotomy-assisted orthodontic treatment on soft tissue clinical parameters in patients with malocclusions with transverse maxillary deficiency. METHODS: The study included 20 generally healthy adult individuals with malocclusion, who underwent a corticotomy-assisted orthodontic treatment in maxilla. During the corticotomy performed after full-thickness flap elevation, only the buccal cortical plate was cut with the use of OTS-7, OTS7-4, OTS7-3 ultrasound tips of the piezosurgery device (Mectron s. p. a., Italy). A clinical examination was performed prior to the corticotomy procedure, then repeated - 3, 6, 9 and 12 months after the procedure. The following parameters were assessed: FMPI (full mouth plaque index), FMBOP (full mouth bleading on probing), PD (probing depth), CAL (clinical attachment level), GR (gingival recession height), RW (recession width), PH (papilla height), PW (papilla width), BS (bone sounding), biotype and KT. RESULTS: There was a statistically significant reduction in PD (mean difference: 0.06; 95% Cl: - 0.33, - 0.18), CAL (mean difference: 0.07; 95% Cl: - 0.33, - 0.19), PH (mean difference: 0.26; 95% Cl: - 0.47, 0.05) and BS (mean difference: 0.13; 95% Cl: - 0.41, - 0.14) after the treatment. Statistically significant changes were also noted in relation to KT (mean difference: 0.17; 95% Cl: - 0.07, 0.27) and biotype (mean difference: 0.07; 95% Cl: 0.26, 0.39), which thickness increased significantly after the treatment. No statistically significant differences were observed in GR, RW and PW. CONCLUSIONS: The corticotomy-assisted orthodontic treatment did not jeopardize the periodontal clinical status in maxilla. There is a need for further studies on a larger number of patient to compare the clinical findings with a control group as well as in patients with conventional orthodontic treatment in a longer follow-up time to find out more about the post-treatment periodontal tissue changes and stability.

Evaluation of concordance between CAD/CAM and clinical positions of abutment shoulder against mucosal margin: an observational study.

BACKGROUND: While working on CAD/CAM-customized abutments, the use of standard impression copings with a circular diameter produces inconsistency within the emergence profile. It may begin with a collapse of the supra-implant mucosa during impression taking, then lead to a computer-generated mismatch of the position and outline of the abutment shoulder, and consequently result in a compromised outcome of anticipated treatment. The aim of the study was to compare the virtual and clinical positions of the abutment shoulder in relation to the mucosal margin after the abutment delivery. METHODS: Conventional open-tray impression takings followed uncovering surgery. Master casts were scanned with a desktop scanner. Clinical examinations took place after abutment's insertion and temporization (T1) and prior to cementation of the definitive crown (T2). The distances between the abutment shoulder and marginal soft tissue were measured intraorally in four aspects and juxtaposed with those on the virtual model. RESULTS: The study evaluated 257 dental implants and CAD/CAM-customized abutments. As T1 and T2 showed, there was a positive correlation between the virtually designed abutment shoulder position and matching clinical location relative to the mucosal margin. In 42.1% of cases, the distance between the mucosal margin and the abutment shoulder did not change. It increased in 36.3% of cases while a decrease occurred in 21.6% of them. CONCLUSIONS: Computer-set position of the abutment shoulder in relation to the mucosal margin can be predictably implemented in clinical practice.

The arachidonate 5-lipoxygenase activating protein gene polymorphism is associated with the risk of scleroderma-related interstitial lung disease: a multicentre European Scleroderma Trials and Research group (EUSTAR) study.

Objectives: The arachidonate 5-lipoxygenase activating protein (ALOX5AP) regulates synthesis of leukotrienes (LTs), which are important mediators of inflammation and connective tissue remodelling. The aim of this study was to evaluate if single nucleotide polymorphisms (SNPs) of ALOX5AP confer risk of SSc and/or SSc-related organ involvement. Methods: Seven SNPs of ALOX5AP (rs17222814, rs17216473, rs10507391, rs4769874, rs9551963, rs9315050 and rs7222842) were genotyped in a cohort of 977 patients with SSc and 558 healthy controls from centres collaborating within the European Scleroderma Trials and Research group. In 22 SSc patients, concentrations of cysteinyl LTs and LT B4 (LTB4) were measured in the supernatants of ionophore-stimulated peripheral blood mononuclear cells (PBMCs) by means of commercially available enzyme immunoassay kits. Results: Significant association was found between rs10507391 polymorphism (T/A) of ALOX5AP and the risk of SSc [odds ratio (OR) 1.27 (95% CI 1.07, 1.50), P < 0.05 vs controls], the presence of SSc-related interstitial lung disease on high-resolution CT of the lungs [OR 1.45 (95% CI 1.17, 1.79), P < 0.05 vs patients without SSc-related interstitial lung disease] as well as with restrictive ventilatory defect [forced vital capacity <70% of predicted; OR 1.51 (95% CI 1.16, 1.97), P < 0.05 vs SSc patients without pulmonary restriction]. PBMCs from SSc carriers of rs10507391 allele A synthesized greater amounts of cysteinyl LTs as compared with SSc patients with rs10507391 TT genotype ( P < 0.05). Synthesis of LTB4 did not differ significantly between the two groups. Conclusion: The results of our study indicate that the genetic variants of ALOX5AP might play a role in the development of SSc-related pulmonary fibrosis.

Time-lapse imaging of cleavage divisions in embryo quality assessment.

In vitro fertilization (IVF) is one of the most important procedures for treating infertility. As several embryos are usually produced in a single IVF cycle, it is crucial to select only the most viable ones for transfer to the patient. Morphokinetics, i.e. analysis of the dynamics of cleavage divisions and processes such as compaction and cavitation, has provided both biologists and clinicians with a new set of data regarding embryonic behaviour during preimplantation development and its association with embryo quality. In the current review, we focus on biological significance of morphokinetic parameters and show how they can be used to predict a reproductive outcome. We also explain the statistics behind the predictive algorithms and discuss the future perspectives of morphokinetics.

Association between Vitamin D Receptor Polymorphism and Serum Vitamin D Levels in Children with Low-Energy Fractures.

OBJECTIVE: Fractures of bones, especially forearm fractures, are very common in children and their number is increasing. This study was designed to determine the impact of vitamin D serum levels and vitamin D receptor (VDR) polymorphisms on the occurrence of low-energy fractures in children. METHODS: The study group consisted of 100 children with clinically relevant bone fractures and a control group consisted of 127 children without fractures. Total vitamin D [25(OH)D3 plus 25(OH)D2] serum concentrations were evaluated in every patient. Genotypes for 4 restriction fragment length polymorphisms of the vitamin D receptor gene (FokI, ApaI, TaqI, and BsmI) were determined by standard polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) techniques. RESULTS: Differences in concentrations of vitamin D were observed between the group with bone fractures (median = 12 ng/ml) and the control group (median = 16 ng/ml; p = 0.000044). Higher levels of vitamin D reduced the risk of fracture by 1.06 times (p = 0.0005). No impact of particular VDR polymorphism on the occurrence of low-energy fractures in children was detected. However, there were significant differences in the prevalence of FokI polymorphism genotypes between the fracture and control groups (p = 0.05). Furthermore, the recessive "aa" genotype of ApaI polymorphism and the dominant "TT" genotype of TaqI polymorphism were associated with higher levels of vitamin D (p = 0.005 and p = 0.036, respectively). CONCLUSIONS: Vitamin D deficiency is an independent risk factor for fractures in children. ApaI polymorphism recessive "aa" and TaqI polymorphism dominant "TT" genotypes are associated with higher levels of vitamin D in serum.

Akt and 14-3-3 control a PACS-2 homeostatic switch that integrates membrane traffic with TRAIL-induced apoptosis.

TRAIL selectively kills diseased cells in vivo, spurring interest in this death ligand as a potential therapeutic. However, many cancer cells are resistant to TRAIL, suggesting the mechanism mediating TRAIL-induced apoptosis is complex. Here we identify PACS-2 as an essential TRAIL effector, required for killing tumor cells in vitro and virally infected hepatocytes in vivo. PACS-2 is phosphorylated at Ser437 in vivo, and pharmacologic and genetic studies demonstrate Akt is an in vivo Ser437 kinase. Akt cooperates with 14-3-3 to regulate the homeostatic and apoptotic properties of PACS-2 that mediate TRAIL action. Phosphorylated Ser437 binds 14-3-3 with high affinity, which represses PACS-2 apoptotic activity and is required for PACS-2 to mediate trafficking of membrane cargo. TRAIL triggers dephosphorylation of Ser437, reprogramming PACS-2 to promote apoptosis. Together, these studies identify the phosphorylation state of PACS-2 Ser437 as a molecular switch that integrates cellular homeostasis with TRAIL-induced apoptosis.

Effect of vitrification on the zona pellucida hardening and follistatin and cathepsin B genes expression and developmental competence of in vitro matured bovine oocytes.

The purpose of our study was to assess the effect of vitrification with or without the presence of calcium in the vitrification solution on the: 1) diameter of oocytes and thickness of the zona pellucida, 2) zona pellucida hardening, 3) expression of mRNA follistatin (FST) and cathepsin B (CTSB) in oocytes and 4) developmental competence of embryos derived from in vitro matured and vitrified oocytes. The results of our study demonstrate, that vitrification did not alter thickness of the zona pellucida and diameter of the oocytes, however it triggered hardening of the zona pellucida. The presence of calcium in the vitrification solutions intensified hardening of zona in immature and mature oocytes (P < 0.04, P < 0.001, respectively) and provoked increased mRNA FST expression in oocytes matured in vitro compared to immature oocytes (P < 0.01) and those vitrified without calcium (P < 0.004). CTSB mRNA expression was increased in immature oocytes and oocytes vitrified with calcium compare to mature oocytes (P < 0.02). The developmental potential of vitrified oocytes was impaired compared to non-vitrified oocytes, being more evident in oocytes vitrified with calcium. In summary, vitrification did not change the oocyte diameter and thickness of the zona pellucida and expression of FST and CTSB mRNA. It diminished developmental potential of the vitrified oocytes. The presence of calcium in the vitrification solutions increased hardening of zona pellucida as well as affected the level of FST and CTSB mRNA in oocytes and developmental potential of these oocytes.

The Effect of Nonsurgical Periodontal Therapy on the Level of Human Neutrophil Peptides 1-3 in Patients with Aggressive Periodontitis.

PURPOSE: To assess the presence of HNP1-3 in the gingival crevicular fluid (GCF) of patients suffering from aggressive periodontitis before and after nonsurgical periodontal therapy. MATERIALS AND METHODS: Twenty patients, each with generalised aggressive periodontitis (GAP) were included in the study. After periodontal examination, one site with a probing depth (PD) ≥ 4 mm was selected. Patients received nonsurgical treatment (scaling and root planing [SRP]) with additional administration of systemic antibiotic therapy (amoxicillin 375 mg three times daily + metronidazole 250 mg three times daily for 7 days). Prior to therapy and 3 and 6 months after, the following parameters were evaluated from the same site: PD, gingival recession (GR), clinical attachment level (CAL), plaque index (PI), bleeding on probing (BOP), sulcus fluid flow rate (SFFR). The level of HNP1-3 in GCF was determined by means of a commercially available ELISA kit. RESULTS: Compared to baseline, the level of HNP 1-3 did not show statistically significant differences at 3 and 6 months. The evaluated clinical parameters and SFFR showed statistically significant decreases compared to baseline. At 6 months, PD (median) decreased from 7 to 3.5 and CAL (median) decreased from 7 to 4. CONCLUSION: In patients with GAP, nonsurgical periodontal therapy in conjunction with systemic administration of amoxicillin and metronidazole had no effect on the level of HNP1-3 in GCF.

Leukotriene production profiles and actions in the bovine endometrium during the oestrous cycle.

We have previously shown the influence of leukotrienes (LTs) on reproductive functions in vivo: LTB4 is luteotrophic and supports corpus luteum function inducing PGE2 and progesterone (P4) secretion, whereas LTC4 is luteolytic and stimulates PGF2α secretion in cattle. The aim of this study was to examine expression and production profiles of LTs and their actions in the endometrium. LT receptors (LTB4R for LTB4 and CysLTR2 for LTC4), 5-lipoxygenase (LO), 12-LO synthase (LTCS) and LTA4 hydrolase (LTAH) mRNA and protein expression, as well as LT production were measured in bovine endometrial tissue during the luteal phases of the oestrous cycle. The action of LTs on uterine function was studied by measuring the level of PGs after stimulating uterine slices with LTs on Days 8-10 of the cycle. Expression of 5-LO and LTB4R mRNA and protein were highest on Days 2-4 of the cycle, while CysLTR2 and LTCS were highest on Days 16-18 (P<0.05). LTB4 concentration was highest on Days 2-4 of the cycle, whereas the greatest LTC4 level was on Days 16-18 (P<0.05). Both LTB4 and C4 increased the content of PGE2 and F2α in endometrial slices at a dose of 10(-7)M (P<0.05). In summary, mRNA expression and activation of receptors for LTB4 and production occur in the first part of the cycle, whereas LTC4 and its receptors predominate at the end of the cycle. The 12-LO and 5-LO pathways are complementary routes of LT production in the bovine uterus.

Serum irisin and its regulation by hyperinsulinemia in women with polycystic ovary syndrome.

Irisin is an adipokine/myokine which could be connected with insulin sensitivity. Polycystic ovary syndrome (PCOS) is characterized by oligo- or anovulation, polycystic ovary, hyperandrogenism and insulin resistance. The aim of the present study was to determine the relationship between serum irisin concentration and insulin sensitivity (Mffm) as well as the effect of insulin infusion on circulating irisin levels in PCOS women as compared with healthy controls. Seventy seven women were enrolled in the study - 57 with PCOS and 20 healthy controls matched for BMI and age. Hyperinsulinemic euglycemic clamps were performed in all of the study participants. The serum concentrations of irisin at baseline and after the clamp, as well as changes of serum irisin concentration in response to insulin supplied during the clamp (Δ irisin), were estimated. The mean serum concentrations of irisin at baseline and after hyperinsulinemia were higher in PCOS women in comparison to the control group (p=0.01; p=0.006, respectively). Insulin infusion resulted in a decrease of serum irisin concentration only in the PCOS group (p=0.007). In the control group, Δ irisin positively correlated with Mffm (r=0.56, p=0.009). In the entire group, multiple regression analysis showed that Δ irisin (ß=0.70, p=0.0002), FFAs 60' during the clamp study (ß=-0.22, p=0.01), SHBG (ß=0.54, p<0.0001) and the interaction between Δ irisin and PCOS (ß=-0.67, p=0.0004) were significantly associated with Mffm. The higher serum irisin concentrations at baseline and in response to insulin infusion might be secondary to insulin resistant conditions in PCOS women.

Do morphokinetic data sets inform pregnancy potential?

PURPOSE: The aim of this study was to create a model to predict the implantation of transferred embryos based on information contained in the morphokinetic parameters of time-lapse monitoring. METHODS: An analysis of time-lapse recordings of 410 embryos transferred in 343 cycles of in vitro fertilization (IVF) treatment was performed. The study was conducted between June 2012 and November 2014. For each embryo, the following data were collected: the duration of time from the intracytoplasmic sperm injection (ICSI) procedure to further division for two, three, four, and five blastomeres, time intervals between successive divisions, and the level of fragmentation assessed in successive time-points. Principal component analysis (PCA) and logistic regression were used to create a predictive model. RESULTS: Based on the results of principal component analysis and logistic regression analysis, a predictive equation was constructed. Statistically significant differences (p < 0.001) in the size of the created parameter between the implanted group (the median value: Me = -5.18 and quartiles: Q 1= -5.61; Q 3 = -4.79) and the non-implanted group (Me = -5.69, Q 1 = -6.34; Q 3 = -5.16) were found. A receiver operating characteristic (ROC) curve constructed for the considered model showed the good quality of this predictive equation. The area under the ROC curve was AUC = 0.70 with a 95% confidence interval (0.64, 0.75). The presented model has been validated on an independent data set, illustrating that the model is reliable and repeatable. CONCLUSIONS: Morphokinetic parameters contain information useful in the process of creating pregnancy prediction models. However, embryo quality is not the only factor responsible for implantation, and, thus, the power of prediction of the considered model is not as high as in models for blastocyst formation. Nevertheless, as illustrated by the results of this study, the application of advanced data-mining methods in reproductive medicine allows one to create more accurate and useful models.