The unique monoclonal antibodies and immunochemical assay for comprehensive determination of the cell-bound and soluble HER2 in different biological samples.

The expression of the HER2 (human epidermal growth factor receptor 2) protein in cancer cells is a well-established cancer marker used for diagnostic and therapeutic purposes in modern treatment protocols, especially in breast cancer. The gold-standard immunohistochemical diagnostic methods with the specific anti-HER2 antibodies are utilized in the clinic to measure expression level of the membrane-bound receptor. However, a soluble extracellular domain (ECD) of HER2 is released to the extracellular matrix, thus the blood assays for HER2 measurements present an attractive way for HER2 level determination. There is a need for accurate and validated assays that can be used to correlate the concentration of the circulating HER2 protein with disease clinical manifestations. Here we describe two monoclonal antibodies binding HER2 with a unique sequence of the complementarity-determining regions that recognize HER2 ECD. Development and validation of the sandwich enzyme-linked immunosorbent assay (ELISA) for quantification of the soluble HER2 in a variety of biological samples is also presented. The assay provides HER2 quantitation within a concentrations range from 1.56 to 100 ng/ml with sensitivity at the level of 0.5 ng/ml that meets the expectations for measurements of HER2 in the blood and tumor tissue samples. The method presents satisfactory intra- and inter-assay precision and accuracy for immunochemical quantification of biomarkers in biological samples. The utility of the generated monoclonal anti-HER2 antibodies has been confirmed for use in the precise measurement of HER2 (both cell-bound and soluble) in several types of biological material, including serum, solid tumor tissue, and cell culture medium. Additionally, the developed immunochemical tools have a potential for HER2 detection, not only in a wide range of sample types but also independently of the sample storage/pre-processing, allowing for comprehensive HER2 analysis in tissue (IHC), cultured cells (immunofluorescence) and blood (ELISA).

Proteins in human body fluids contain in vivo antigen analog of the melibiose-derived glycation product: MAGE.

Melibiose-derived AGE (MAGE) is an advanced glycation end-product formed in vitro in anhydrous conditions on proteins and protein-free amino acids during glycation with melibiose. Our previous studies revealed the presence of MAGE antigen in the human body and tissues of several other species, including muscles, fat, extracellular matrix, and blood. MAGE is also antigenic and induces generation of anti-MAGE antibody. The aim of this paper was to identify the proteins modified by MAGE present in human body fluids, such as serum, plasma, and peritoneal fluids. The protein-bound MAGE formed in vivo has been isolated from human blood using affinity chromatography on the resin with an immobilized anti-MAGE monoclonal antibody. Using mass spectrometry and immunochemistry it has been established that MAGE epitope is present on several human blood proteins including serum albumin, IgG, and IgA. In serum of diabetic patients, mainly the albumin and IgG were modified by MAGE, while in healthy subjects IgG and IgA carried this modification, suggesting the novel AGE can impact protein structure, contribute to auto-immunogenicity, and affect function of immunoglobulins. Some proteins in peritoneal fluid from cancer patients modified with MAGE were also observed and it indicates a potential role of MAGE in cancer.

Glycoproteins Presenting Galactose and N-Acetylgalactosamine in Human Seminal Plasma as Potential Players Involved in Immune Modulation in the Fertilization Process.

In light of recent research, there is increasing evidence showing that extracellular semen components have a significant impact on the immune reaction of the female partner, leading to the tolerogenic response enabling the embryo development and implantation as well as further progress of healthy pregnancy. Seminal plasma glycoproteins are rich in the unique immunomodulatory glycoepitopes that may serve as ligands for endogenous lectins that decorate the surface of immune cells. Such interaction may be involved in modulation of the maternal immune response. Among immunomodulatory glycans, Lewis type antigens have been of interest for at least two decades, while the importance of T/Tn antigens and related structures is still far from understanding. In the current work, we applied two plant lectins capable of distinguishing glycoepitopes with terminal GalNAc and Gal to identify glycoproteins that are their efficient carriers. By means of lectin blotting and lectin affinity chromatography followed by LC-MS, we identified lactotransferrin, prolactin inducible protein as well as fibronectin and semenogelins 1 and 2 as lectin-reactive. Net-O-glycosylation analysis results indicated that the latter three may actually carry T and/or Tn antigens, while in the case of prolactin inducible protein and lactotransferrin LacdiNAc and lactosamine glycoepitopes were more probable. STRING bioinformatics analysis linked the identified glycoproteins in the close network, indicating their involvement in immune (partially innate) processes. Overall, our research revealed potential seminal plasma ligands for endogenous Gal/GalNAc specific lectins with a possible role in modulation of maternal immune response during fertilization.

Immunomodulatory Factors in Primary Endometrial Cell Cultures Isolated from Cancer and Noncancerous Human Tissue-Focus on RAGE and IDO1.

BACKGROUND: Immune modulatory factors like indoleamine 2,3-dioxygenase 1 (IDO1) generating kynurenine (Kyn) and receptor for advanced glycation end-products (RAGE) contribute to endometrial and cancer microenvironment. Using adequate experimental models is needed to learn about the significance of these molecular factors in endometrial biology. In this paper we study IDO1 activity and RAGE expression in the in vitro cultured primary human endometrial cells derived from cancerous and noncancerous tissue. METHODS: The generated primary cell cultures from cancer and noncancerous endometrial tissues were characterized using immunofluorescence and Western Blot for expression of endometrial and cancer markers. IDO1 activity was studied by Kyn quantification with High Performance Liquid Chromatography with Diode Array Detector. RESULTS: The primary cultures of endometrial cells were obtained with 80% success rate and no major genetic aberrations. The cells retained in vitro expression of markers (mucin MUC1 and HER2) or immunomodulatory factors (RAGE and IDO1). Increased Kyn secretion was associated with cancer endometrial cell culture in contrast to the control one. CONCLUSIONS: Primary endometrial cells express immune modulatory factors RAGE and IDO1 in vitro associated with cancer phenotype of endometrium.

Evaluation of the health behaviors of women in multiple pregnancies - a preliminary study.

OBJECTIVES: The aim of the paper was to assess which health behavior indicators were characteristic for women with a multiple pregnancy. MATERIAL AND METHOD: The study involved 35 women in multiple pregnancies. The inclusion criteria were: the completion of the 22nd week of pregnancy (22 Hbd, i.e., 22 weeks and 1/7) and the consent of the women to participate in the study. We used an original questionnaire devised by us specifically for our study, as well as a standardized research tool, Juczynski's Health Behavior Inventory questionnaire, for the assessment of the pregnant women's health behaviors. RESULTS: The overall rate of health behaviors was high and was on average 93.9 ± 10.8 points (7 sten scores). Indicators of the health behaviors of women in multiple pregnancies were high across all categories. However, the highest rate was observed in preventive behaviors. CONCLUSIONS: The overall rate of health behaviors of women with a multiple pregnancy and all the individual behavior category indicators were high. However, the highest rate was for preventive behaviors. The indicator values of the health behaviors of the pregnant women in the study were not dependent on the variables adopted in this paper.

Anti-Müllerian hormone: structure, properties and appliance.

Anti-Müllerian hormone (AMH) is a glycoprotein produced by the granulosa cells of preantral and small antral follicles. AMH concentrations reflect ovarian physiology with high precision, thus serving as a more sensitive marker of the ovarian reserve than the chronological age. This hormone plays a role in the pathogenesis of menstrual disorders and fertility in obesity and polycystic ovary syndrome. The evaluation of AMH may also be useful in the diagnosis or the monitoring therapy of granulosa cells ovarian tumors.

Anti-Müllerian hormone: a critical factor for female fertility and reproductive health.

Anti-Müllerian hormone (AMH) is a glycoprotein produced by the granulosa cells of preantral and small antral follicles. AMH concentrations reflect ovarian physiology with high precision, thus serving as a more sensitive marker of the ovarian re-serve than chronological age. This hormone plays a role in the pathogenesis of menstrual disorders and fertility in both obesity and polycystic ovary syndrome. The evaluation of AMH may also be useful in diagnosing or monitoring therapy of granulosa cell ovarian tumors.

[Awaiting elective gynecological surgery--opinions of patients]. / Oczekiwanie na planowana operace ginekologiczna w ocenie kobiet.

PURPOSE: To establish if and to what extend women find the time of waiting for an elective gynecological operation difficult, how it is manifested and what the conditioning factors are. MATERIAL AND METHOD: The investigation was carried out in the group of 272 women who during two months of 2008 were admitted to one of the four selected hospitals for an elective gynecological operation. A questionnaire has been designed solely for the purpose of the pilot study 25 women participated in the study. RESULTS: The respondents reported 717 concerns that occurred within the period of waiting for the surgery; the most frequent among them were fear of being diagnosed with cancer (27.6%), postoperative pain (26.7%) postoperative complications (19.1%). More than half of the respondents (58.5%) waited for the surgery for 2-3 months or longer. Every fifth woman complained about problems with being admitted to hospital and every fourth woman expected emotional support from medical staff. 91.2% respondents had searched for additional information about their disease. CONCLUSIONS: 1. Waiting for elective gynecological operation may create a difficult situation for many women what was confirmed by a vast number and nature of their concerns, frequent expectations of emotional support and search for additional information about their disease. 2. Problems with being admitted to a health care institution and long period of waiting for the operation (> 1 month) are quite common and these factors may contribute to further worsening of the quality of patients' life during the preoperative period. 3. Demographic parameters assumed for the purpose of this analysis do not differentiate the parameters tested, with the exception of the place of inhabitance factor that turned out to be associated with the expectations of emotional support. 4. The degree of hospital reference significantly differentiates tested parameters.

Bipolar affective disorder in a male with a deletion of Y chromosome -- a case report.

We report on a 25-year-old male with bipolar disorder, dysmorphic features and a deletion of the long arm of Y chromosome. A potential association between sex chromosome abnormalities and a susceptibility to major psychiatric disorders has been documented. However there have been very few reports on the coincidence of Y chromosome aberrations with bipolar disorder. Cytogenetic studies have contributed to the identification of several disease genes. Karyotyping of patients with bipolar disorder in order to identify candidate regions for linkage studies has been recommended.

Activity of superoxide dismutase and glutathione peroxidase in human semen in normozoospermia and spermatopathy.

The aim of the study was to assess the levels of antioxidative enzymes--superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the whole semen of men treated for infertility. The first group consisted of the patients with normozoospermia; the second--included the men with oligozoospermia. According to sperm motility, the patients were divided into the third group--motility higher than 50% and the fourth group--motility lower than 50%. The study showed that the levels of both enzymes were higher in men with oligozoospermia than those with normozoospermia. It was also found that the activities of SOD and GSH-Px were higher in men with low motility(< 50%) compared to high motility patients. The results confirm that the activity of superoxide dismutase and glutathione peroxidase is related to the parameters of human semen assessed according to the WHO standards.