Low-level 45,X/46,XX mosaicism is not associated with congenital heart disease and thoracic aorta dilatation:prospective magnetic resonance imaging and ultrasound study.

OBJECTIVE: To establish the prevalence of risk factors for aortic dissection, such as bicuspid aortic valve, aortic coarctation and ascending aorta dilatation, in women with low-level 45,X/46,XX mosaicism undergoing an in-vitro fertilization (IVF) procedure. METHODS: The study group comprised 25 women with low-level 45,X/46,XX mosaicism (ranging from 3.3% to 10.0%) who were referred to two reproductive medicine units between 2009 and 2013 because of infertility and who underwent subsequent karyotyping. In accordance with the recommendation of the Practice Committee of the American Society for Reproductive Medicine for patients with Turner syndrome (TS), prior to the IVF procedure, all women underwent careful cardiovascular screening for congenital heart disease and thoracic aorta dilatation, including standard cardiac examination, echocardiography and non-contrast cardiac magnetic resonance imaging. Aortic size index (ASI, diameter of the ascending aorta normalized to body surface area) and the prevalence of coarctation of the aorta and of bicuspid aortic valve were compared with findings previously reported in women with TS and the general population. RESULTS: Bicuspid aortic valve without any stenosis or regurgitation was found in one woman in the study group with low-level 45,X/46,XX mosaicism, a statistically significantly lower prevalence of bicuspid aortic valve than that reported in women with TS. Aortic coarctation was not identified in any individual. The ASI was below the 95th percentile in all cases and the mean value was significantly lower than the mean reference values for both the general population and women with TS. CONCLUSION: Compared with the general population, the prevalence of risk factors for aortic dissection was not found to be higher in women with low-level 45,X/46,XX mosaicism without any noticeable features except infertility.

[Prevalence of Chlamydia trachomatis infection in patients treated for infertility]. / Výskyt Chlamydia trachomatis pri lécbe neplodnosti.

AIM OF THE STUDY: To determine the prevalence of Chlamydia trachomatis infection in patients treated for infertility. STUDY TYPE: A retrospective analysis. SETTING: Fertimed, infertility treatment center, Olomouc. METHOD: At Fertimed, we used DNA detection of Chlamydia trachomatis by the PCR method of the company GeneProof to examine, between 2009-2011, 785 women undergoing one of the infertility treatment methods and their 113 partners. In the second group, we examined 121 oocyte donors and 30 men before sperm donation. We appraised the frequency of Chlamydia trachomatis detection in the specific groups and the clinical impact of the infection on the female reproductive organs. RESULTS: In the group of women treated for infertility, we detected 20 (2.5%) women with an active infection. After treatment, 9 of them underwent an examination of Fallopian tube patency using the UTHL (ultrasonographically guided transvaginal hydrolaparoscopy) method. In 7 cases, we indicated a bilateral salpingectomy due to a sactosalpinx and in one case severe pelvic adhesions were found (88.9%), and in one patient, the result was normal. In the control group of 43 PCR-negative women who were examined for Fallopian tube patency, 9.3% rate of tubal pathology was found (p<0.001). In the oocyte donor group, we detected the presence of Chlamydia trachomatis in 12 (9.9%) women, and in the sperm donor group, in 7.6% men. Treatment with 500 mg of Sumamed (azithromycin), given in 3 doses, was successful in all of the positive patients. CONCLUSION: We found that Chlamydia trachomatis detection was lower in the women treated for infertility than in the female donor group. Women with a confirmed infection had a high prevalence of inflammatory changes in the Fallopian tubes compared with women devoid of a confirmed infection. The treatment with azithromycin is effective.

Visfatin is secreted into the breast milk and is correlated with weight changes of the infant after the birth.

INTRODUCTION: Visfatin is a recently identified adipokine with numerous metabolic and immunoregulatory properties that has been implicated in the regulation of the white adipose tissue (WAT) and significant changes in visfatin levels were reported during pregnancy. The aim of the study was to investigate dynamics of visfatin levels in maternal serum and human breast milk during a 180-d period after the delivery. MATERIALS AND METHODS: : Breast milk and venous blood samples were obtained from 24 healthy lactating women with uncomplicated, physiological pregnancy and appropriate-for-gestational age neonates and serum-milk sample duos were collected at the time of birth, at the 1-3, 12-14, 28-30, 88-90 and 178-180 postpartum. RESULTS: Our study demonstrates that (1) visfatin is abundantly secreted into breast milk in humans, reaching approx. 100× higher concentrations compared to maternal serum; (2) visfatin concentrations in maternal serum show significant variations after the delivery and (3) visfatin concentration in colostrum could be used for prediction of the subsequent weight development (less/more severe weight loss during first 3 days after the birth) of the infant. DISCUSSION: Our data suggest that visfatin could play an important role in regulation of adiposity of the infant after the birth.

Relationship of resistin levels with endometrial cancer risk.

Cancer of endometrium (CAE) is the most common gynecologic malignancy in industrialized nations. Increased resistin levels, an adipocytokine produced by adipose tissue and macrophages, have been considered as a risk factor in gastric, colon and breast cancer, recently. No studies associating resistin levels with endometrial cancer have been done so far. The purpose of this case-control study was to determine the relationship between serum circulating resistin levels and resistin gene -420C>G (rs3219175) variant in endometrial cancer patients. 37 Caucasian female patients and 39 healthy controls were enrolled in this study. Difference in resistin levels between age and BMI matched patients group (mean 24.2 ng/ml) and control subjects (mean 10.1 ng/ml) were statistically significant (p <001). We also determined single nucleotide polymorphism -420C>G (rs3219175) within resistin gene and no significant association between resistin levels and investigated polymorphism was found. Furthermore, no significant association between higher resistin levels and diabetes mellitus 2, body mass index, smoking or age have been observed within studied groups. To our knowledge, this is the first study examining the relationship between serum resistin levels and endometrial cancer and our results show, that patients with endometrial cancer have significantly increased circulating levels of resistin compared to control subjects.

Leptin--2548 g/A polymorphism in endometrial cancer.

BACKGROUND: Previously, the polymorphism-2548 G/A within the promoter of the leptin (LEP) gene was reported to be associated with overweight and obesity, the factors significantly associated to increased endometrial cancer risk. Leptin has been described to play an important role in signal transduction in endometrial cancer cells indicating that leptin promotes endometrial cancer growth and invasiveness and implicating the JAK/STAT and AKT pathways as critical mediators of leptin action. The aim of the study was to investigate the possible associations of LEP-2548 G/A polymorphism with endometrial cancer and its related traits. DESIGN: Using PCR with following restriction analysis, we studied 67 endometrial cancer cases (mean age 64.3 +/- 10.3 years) that were enrolled in the study along with 67 controls matched for age, BMI and ethnic origin (mean age 62.1 +/-9.8 years); an additional cohort of 543 healthy individual was recruited to investigate the general population frequencies. RESULTS: The present study revealed no significant differences between the genotypes or alleles of investigated polymorphism for endometrial cancer risk or its related traits (age of menarche, menopause, number of spontaneous abortions in personal history or waiting time till the onset of the disease) among the groups, thus indicating that the genetic variants of LEP-2548 G/A is not a relevant marker of endometrial cancer risk in this Czech population. CONCLUSIONS: To conclude, the polymorphism LEP-2548 G/A doesn't seem to represent a major genetic marker for endometrial cancer in the studied Czech population; however, it was associated with obesity, which finding is in accordance with previous reports.

[Level of CA125 and hemoglobin as prognostic factors ovarian cancer]. / Vliv hodnoty CA125 a hemoglobinu na prognózu ovariálního karcinomu.

AIM OF THE STUDY: Aim of the study was to investigate connection between the level CA125, level of haemoglobin and emergence of recurrence and survival time for patients with ovarian cancer. MATERIAL AND METHODS: This concerns of retrospective study in which total 53 patients with histology confirmed ovarian cancer were included. These patients were diagnosed and subsequently treated in the GPK FN MU Brno during 2000-2002 period. RESULTS: The level of CA125 in our study appeared to be statistically significant prognostic factor (beta = 0.44, p = 0.02) for survival. Signification of the plasma level of haemoglobin to be any prognostic factor was not proved in this study. Among other assessed factors influencing survival time of the patients was the number of gravidity to be important prognostic factor (beta = -0.96, p = 0.002). Survival time was shorter for patients with less number of pregnancies. CONCLUSION: Ovarian cancer is tumour with high mortality. That is why it is necessary to look for new markers, which would contribute to earlier diagnostic of this disease, and which could influence its prognosis.

[Cholecystectomy--the risk factors for endometrial carcinoma?]. / Cholecystektomie--rizikový faktor karcinomu endometria?

AIM: The aim of this study was to find out if the incidence of cholecystectomy is more frequent in patiens with endometrial carcinoma and if the incidence of the anamnestic information concerning cholecystectomy in patiens with endometrial carcinoma influences the course and the result of the treatment of the said tumorous disease. TYPE OF STUDY: Retrospective study. METHODS: 470 females with endometrial carcinoma were entered into the study, the group was divided into the patients who have had cholecystectomy and those who have not had it. There was a control group of 370 females. All the said patients were diagnosed and subsequently treated at the Department of Gynecology and Obstetrics Masaryk University affiliated Hospital, Brno between January 2004 and December 2006. RESULTS: We have not proved any statistically significant difference concerning the origin of the endometrial carcinoma after a performed cholecystectomy. There was no difference in grading as well as the stage according to the FIGO standards. Nonetheless, we can say that the patients with endometrial carcinoma and cholecystectomy have significantly higher BMI. CONCLUSION: The relation between cholecystectomy and the endometrial carcinoma can be influenced by various factors. However, we can assume that the pathological processes of cholelithiasis are pathophysiologically connected with the development of changes, which fall within the category of hormonal risk factors for the development of endometrial carcinoma.

[Insertion-deletion polymorphism in the gene for angiotensin-converning enzyme (I/D ACE) in pregnant women with gestational diabetes]. / Inzercne-delecní polymorfismus v genu pro angiotensin-konvertující enzym (I/D ACE) u tehotných s gestacním diabetem.

AIM OF THE STUDY: The aim of the study was to investigate possible associations of the Angiotensin-converting enzyme insertion/deletion (I/D ACE) polymorphism with gestational diabetes mellitus occurrence. METHODS: Number of 53 healthy pregnant women (controls) and 48 women with gestational diabetes mellitus were included in the study. The woman patients were hospitalised in the Clinic of Obstetrics and Gynaecology, Masaryk University Affiliated hospital Brno during the period 10/2004 - 10/2005. In all cases there was the spontaneous one baby pregnancy. The procedure was obtaining the peripheral blood samples from women of those two groups and isolation of their DNA using standard technique with the use of proteinase K. I/D ACE genotype of each person was determined using allele specific primers. RESULTS: We did not prove significant association neither of the genotype distribution (P(g)) P(g) = 0.115 nor allele frequency (P(a)) P(a) = 0.873 between controls and women with gestational diabetes mellitus. The significant differences of genotype distributions (P(g) = 0.03) were observed in women with two and more labours in anamnesis. Borderline significant differences of genotype distributions (P(g) = 0.08) were found in women with one and more abortions in anamnesis. CONCLUSIONS: According to our results the variability in ACE gene can be taken as factor contributing to manifestation of gestational diabetes mellitus. According to results of our study I/D ACE polymorphism can't be taken as genetic marker of gestational diabetes mellitus.

Spermadhesins: a new protein family. Facts, hypotheses and perspectives.

Spermadhesins are a novel family of secretory proteins expressed in the male genital tract of pig, horse and bull. They are major products of the seminal plasma and have been found to be peripherally associated to the sperm surface. The structure and function of spermadhesins have been thoroughly investigated in the pig, which exhibits the greatest diversity of members: AWN, AQN-1, AQN-2, PSP-I and PSP-II and its glycosylated isoforms. They are multifunctional proteins showing a range of ligand-binding abilities, e.g. carbohydrates, sulfated glycosaminoglycans, phospholipids and protease inhibitors, suggesting that they may be involved in different steps of fertilization. Isolated porcine spermadhesins bind the zona pellucida glycoproteins in a cation-dependent manner with a Kd in a low micromolar range, and AWN, AQN-1 and AQN-3 display similar binding affinity for glycoproteins containing Gal beta(1-3)-GalNAc and Gal beta(1-4)-GlcNAc sequences in O-linked and N-linked oligosaccharides, respectively. During sperm passage through the epididymis AQN-3 and AWN have been shown to bind tightly to the sperm surface by interaction with the phospholipids of the membrane bilayer. At ejaculation the spermadhesins form a protective coat around the sensitive acrosomal region of the sperm head, thus possibly preventing premature acrosome reaction. During in vitro capacitation most of these aggregated sperm adhesins are lost, with the exception of phospholipid-bound spermadhesins. AWN and AQN-3 may now serve as a primary receptor for the oocyte zona pellucida, thus contributing to initial binding and recognition between sperm and egg. The amino acid sequence of spermadhesins does not show any discernible similarity with known carbohydrate recognition domains (CRD). However, they belong to the superfamily of proteins with a CUB domain with a predicted all-beta structure. The crystal structure of the heterodimeric complex of the spermadhesins PSP-I/PSP-II has been solved, showing that the overall structure of both spermadhesins consists of a beta-sandwich with five (parallel and antiparallel) beta-strands. It is the first three-dimensional structure of a zona pellucida-binding protein and reveals the architecture of the CUB domain. The spermadhesins represent a novel class of lectins that may be involved in sequential steps of fertilization, at least in the pig.

Mapping the heparin-binding domain of boar spermadhesins.

Boar spermadhesins are a group of seminal plasma, heparin-binding proteins which appear to be involved in sperm capacitation and gamete interaction. Using a proteolytic protection assay we have identified regions of AQN-1, AQN-3, PSP-I and AWN which remain attached to a heparin-Sepharose column following in-column digestion of bound spermadhesins with chymotrypsin and elastase. In addition, the complete amino acid sequence of spermadhesin AWN was synthesized as overlapping peptides, and their ability to bind to a heparin-Sepharose column and to inhibit the interaction of soluble heparin with purified ELISA plate-coated AWN was tested. Both approaches gave similar results and as a whole showed that different regions of AWN may converge in its tertiary structure to form a composite heparin-binding site. The conformational heparin-binding surface resides on the GFCC'C'' face of the proposed structural model for AWN and is in an opposite location to the carbohydrate-binding region of the spermadhesin.

Boar spermadhesin AWN-1. Oligosaccharide and zona pellucida binding characteristics.

We have analyzed the oligosaccharide recognition ability of boar spermadhesin AWN-1 using biotinylated glycoproteins with defined carbohydrate chains as probes. Our results show that AWN-1 bound to proteins containing O-linked NeuAc alpha(2-3/6)-Gal beta(1-3)-GalNAc (PDC-109 and fetuin) with a Kd = 0.7 microM. AWN-1 also bound to NeuAc alpha(2-3/6)-Gal beta(1-4)-GlcNAc sequences in N-linked triantennary structures of fetuin, but not to the same oligosaccharide in the diantennary structures of IgG or fibrinogen. The absence of terminal sialic acid decreased fivefold the binding affinity. By competitive ELISA, peptides containing the N-linked oligosaccharide sequence inhibited the binding of the parent glycoprotein to immobilized AWN-1 5-45 times less effectively than those carrying O-linked NeuAc alpha(2-3/6)-Gal beta(1-3)-GalNAc structures. In addition, AWN-1 bound with a Kd = 0.3 microM to solubilized, biotinylated porcine zona pellucida glycoproteins. PDC-109 competed effectively with zona pellucida glycoproteins for AWN binding, whereas fetuin was a poor competitor. On the other hand, AWN epitopes were demonstrated on in vitro capacitated boar spermatozoa which were able to bind to, and penetrate, zona-encased oocytes. These data indicate that spermadhesin AWN-1 may play a role in pig fertilization as a sperm-associated lectin of broad specificity though preferential affinity for certain O-linked oligosaccharide structures of the oocyte's zona pellucida.

Interaction of non-aggregated boar AWN-1 and AQN-3 with phospholipid matrices. A model for coating of spermadhesins to the sperm surface.

Boar spermadhesins are 12-14 kDa lectins which coat the entire acrosomal cap sperm head surface. The large amount of spermadhesins AQN-1, AQN-2, AQN-3, and AWN-1 present on in vitro capacitated spermatozoa (approximately 7 x 10(6) molecules of each spermadhesin per cell) suggested that they may bind to major component(s) of the sperm surface. We have investigated both the aggregation state of spermadhesins in seminal plasma using gel filtration chromatography, and their ability to bind to the major phospholipids of the boar sperm plasma membrane, i.e. phosphorylcholine and phosphorylethanolamine. The bulk (90%) of spermadhesins AQN-3 and AWN-1 were eluted as aggregated proteins (Mr > 50,000) with the void volume of a Sephadex G-50 column; the remaining 10% of the total amount of seminal plasma AWN-1 and AQN-3 were recovered, together with the whole amount of AQN-1 and AQN-2, in a fraction containing low-molecular-mass proteins (Mr 16,000-30,000). None spermadhesin of either gel-filtration fraction bound to a phosphorylcholine affinity matrix. On the other hand, low-molecular-mass (monomeric or dimeric) AQN-3 and AWN-1 were the only spermadhesins retained in a phosphorylethanolamine affinity column. Both AQN-3 and AWN-1 purified from seminal plasma by reverse-phase HPLC retained their lipid-binding capability. In addition, immobilization of AQN-3 and AWN-1 onto a phosphorylethanolamine matrix did not interfere with the ability of the proteins to bind bovine glycoprotein PDC-109, indicating that the structural determinants for the binding lipid and carbohydrates lay on different structural domains.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunolocalization and quantitation of acidic seminal fluid protein (aSFP) in ejaculated, swim-up, and capacitated bull spermatozoa.

Acidic seminal fluid protein (aSFP, 12.9 kDa), a major protein of bull seminal plasma, belongs to the spermadhesin protein family. Boar spermadhesins become bound to the sperm head's surface at ejaculation and are thought to play a role as capacitation factors and/or in gamete recognition and binding. Here, we have investigated the topographical distribution and fate of bovine spermadhesin aSFP during sperm capacitation in order to assess whether aSFP could be involved in similar aspects of the fertilization process as its boar homologous proteins. 5.7 +/- 2.1 x 10(6) molecules/spermatozoa were quantitated on the surface of fresh ejaculated and washed sperm. The binding site of aSFP was restricted to a thin coat at the apical part of the acrosomal cap. The amount of aSFP in swim-up sperm was 1.8 +/- 1.0 x 10(6) molecules/spermatozoa, but decreased dramatically to 22 +/- 10 x 10(3) and to undetectable levels after incubation of sperm for 1.5 h and 18 h, respectively, in capacitation medium. This indicates that the bull spermatozoa surface may be completely depleted of spermadhesin aSFP before spermatozoa reach the surroundings of the investing egg. Therefore, our results suggest that aSFP may act as a decapacitation factor on bull spermatozoa rather than as a zona pellucida binding molecule.

Quantitation of boar spermadhesins in accessory sex gland fluids and on the surface of epididymal, ejaculated and capacitated spermatozoa.

Spermadhesins are multifunctional proteins involved in boar sperm capacitation and gamete recognition. Using anti-AWN antibodies, we have followed the fate of spermadhesin AWN along the maturation and capacitation stages of boar spermatozoa. In addition, the amount of spermadhesins AQN-1, AQN-2, and AQN-3 relative to that of AWN was determined by amino acid analysis after reverse-phase HPLC isolation. Our data show that AWN-1 is the only spermadhesin on the surface of epididymal sperm and that a large amount of AQN-1, AQN-2, AQN-3, AWN-1 and AWN-2 become coated on ejaculated spermatozoa. The number of spermadhesin molecules on ejaculated sperm (12-60 x 10(6)/spermatozoa) is sufficient to form a many-molecules-thick coat over the sperm head. However, 50-75% of the AQN-1, AQN-2, and AQN-3 population, and around 90% of coated AWN (1 + 2) are released from ejaculated sperm during capacitation. This indicates that a large subpopulation of each boar spermadhesin is loosely associated to the sperm surface and may function as decapacitation factors. The remaining spermadhesin molecules, which are tightly bound to the sperm head's surface may play a role as either positive capacitation factors and/or in gamete recognition and binding.

Characterization of AWN-1 glycosylated isoforms helps define the zona pellucida and serine proteinase inhibitor-binding region on boar spermadhesins.

Spermadhesin AWN-1 (14 kDa) belongs to a recently described family of boar sperm surface-associated proteins. AWN-1 is a multifunctional protein which possesses heparin-, serine proteinase inhibitor-, and zona pellucida glycoprotein-binding capability. Therefore it has been implicated in sperm capacitation and sperm-oocyte attachment. Here, we report the characterization of 22-25 kDa isoforms of AWN-1 isolated by heparin-affinity chromatography, which fail to bind to zona pellucida glycoproteins or serine proteinase inhibitors. Our results show that the structure of the high and low molecular mass AWN-1 forms differ in that the former is N-glycosylated at Asp50 and truncated at the C-terminus. The inability of the glycosylated AWN-1 molecules to bind ligands is due solely to the presence of the oligosaccharide moieties, however. This indicates that glycosylation of AWN-1 may modulate its ligand-binding capabilities. On the other hand, the effect of glycosylation on ligand-binding suggests that both the zona pellucida- and the serine proteinase inhibitor binding domain(s) may be located around the glycosylation point.