Krzysztof Cendrowski

Determination of zearalenone and its metabolites in endometrial cancer by coupled separation techniques

This study presents a selective method of isolation of zearalenone (ZON) and its metabolite, α-zearalenol (α-ZOL), in neoplastically changed human tissue by accelerated solvent and ultrasonic extractions using a mixture of acetonitrile/water (84/16% v/v) as the extraction solvent. Extraction effectiveness was determined through the selection of parameters (composition of the solvent mixture, temperature, pressure, number of cycles) with tissue contamination at the level of nanograms per gram. The produced acetonitrile/water extracts were purified, and analytes were enriched in columns packed with homemade molecularly imprinted polymers. Purified extracts were determined by liquid chromatography (LC) coupled with different detection systems (diode array detection - DAD and mass spectrometry - MS) involving the Ascentis RP-Amide as a stationary phase and gradient elution. The combination of UE-MISPE-LC (ultrasonic extraction - molecularly imprinted solid-phase extraction - liquid chromatography) produced high (R ≈ 95–98%) and repeatable (RSD < 3%) recovery values for ZON and α-ZOL.

Analytical procedure for the determination of zearalenone in environmental and biological samples.

The metabolism of zearalenone (ZEA) and analytical methods for determining the presence of ZEA and its metabolites are discussed in this study. Similar to phytoestrogens, solid metaloestrogens, pharmaceuticals, and selected pesticides, ZEA is a substance that displays endocrine activity. ZEA is accumulated in living organisms, and it is capable of contaminating all trophic levels of the food chain, from grain, maize, and other crop plants to human consumers. Zearalenone has a structure similar to that of estrogen (the presence of a macrocyclic lactone ring), it has an affinity for estrogen receptors, and it competes with 17β-estradiol for binding the estrogen receptor in natural pathways. As endocrine disruptors, zearalenone and its metabolites can also contribute to carcinogenic mutations associated with female secondary sex characteristics. The determination of zearalenone and its metabolites in various matrices, first of all biological and environmental samples, poses significant problems. A variety ways of extracting and purifying zearalenone, including liquid-liquid extraction and solid-phase extraction, are described. Furthermore, it describes the possibility of applying a plurality of sensitive and specific instrumental methods, chromatographic techniques (TLC, HPLC, GC) as well as other methods (immunoaffinity chromatography).

Growth factors, silver dressings and negative pressure wound therapy in the management of hard-to-heal postoperative wounds in obstetrics and gynecology: a review

PurposeThe last two decades witnessed the development of numerous innovative regimens for the management of patients with abnormally healing and infected wounds. Growth factors, negative pressure wound therapy (NPWT) and antiseptic dressings containing silver are examples of methods with best documented efficacy, being widely used in the treatment of acute and chronic post-traumatic wounds, burns and ulcers of various etiology. As far as obstetrics and gynecology are concerned, prevention and treatment of infected, hard-to-heal postoperative wounds is of crucial importance. This article reviews the available literature to discuss the possibilities for use, efficacy and cost-effectiveness of growth factors, NPWT and silver dressings in the treatment of difficult-to-heal postsurgical wounds in obstetrics and gynecology.Materials and methodsAn extensive search of the English and Polish literature via PubMed and EMBASE databases was undertaken for articles published between January 1960 and April 30, 2014 to identify articles that described and assessed use, efficacy and cost-effectiveness of growth factors, silver dressings and NPWT in patients with hard-to-heal postoperative wounds following obstetric or gynecological surgery.ConclusionsLiterature review regarding the use of growth factors, NPWT and silver dressings suggests that these methods may play an important role in the management of wounds after invasive obstetric and gynecological procedures. Obese patients, patients after vulvectomy or prior radiation therapy may benefit most, however, due to non-numerous randomized reports, prospective studies on the use of above-mentioned methods in the treatment of postsurgical wounds following obstetric and gynecological interventions are required.

Expression of vascular endothelial growth factor mRNA in human leiomyomas.

Aim. Growth factors play a major role in the pathogenesis of uterine myomas. The aim of this study was to evaluate vascular endothelial growth factor (VEGF) mRNA expression during leiomyoma growth at different phases of the menstrual cycle with RT-PCR. Method. We studied 56 patients: 43 with myomas, 13 with healthy myometriums. In patients with myomas (secretory phase), VEGF expression was 2.82 times higher than in control patients (p < 0.05). In patients with myomas (phase I), VEGF expression was 2.53 times higher (p < 0.05) than in control patients. For all patients with myomas, those who were in menopause had 1.52 times higher VEGF expression than those who menstruated. For patients with healthy myometriums, those who were in menopause had 1.97 times higher VEGF expression than those who menstruated. A comparison of all the patients in menopause revealed that VEGF expression was 2.03-fold higher in those with myomas than in those with healthy myometriums. Conclusion. We observed the highest VEGF mRNA expression in women with myomas who were in menopause. Among menstruating patients, VEGF expression was significantly higher in those with myomas compared to those with a healthy myometrium. This suggested that VEGF may play a significant role in the pathogenesis of uterine myomas.

Chorioamnionitis (ChA) modifies CX3CL1 (fractalkine) production by human amniotic epithelial cells (HAEC) under normoxic and hypoxic conditions

BackgroundChemokine CX3CL1 possesses unique properties, including combined adhesive and chemotactic functions. Human amniotic epithelial cells (HAEC) show expression of CX3CL1 receptor (CX3CR1) and produce CX3CL1 in response to both physiologic and pathologic stimuli. Chorioamnionitis (ChA) is a common complication of pregnancy and labour. ChA is often accompanied by local hypoxia because of the high oxygen consumption at the site of inflammation. We examined comparatively (ChA-complicated vs. normal pregnancy) CX3CR1 expression and the effects of hypoxia, lipopolysaccharide (LPS), and CX3CR1 blockade on CX3CL1 production in HAEC cultured in vitro.MethodsHAEC have been isolated using trypsinization, and cultured under normoxia (20% O2) vs. hypoxia (5% O2). According to the experimental design, LPS (1 μg/ml) and neutralizing anti-CX3CR1 antibodies were added at respective time points. Mean CX3CL1 concentration in the supernatant samples were determined by ELISA. Expression of immunostained CX3CR1 was analyzed using quantitative morphometry.ResultsWe have found that the mean levels of CX3CL1 and CX3CR1 expression were remarkably (p < 0.05) higher in ChA, compared to normal pregnancy. Significantly increased expression of CX3CR1 was observed in ChA during both normoxia and hypoxia. Hypoxia exposure produced decrease in the mean concentration of CX3CL1 in both groups, however this reduction was stronger in normal pregnancy. In normoxia, LPS-evoked rise in the mean concentration of CX3CL1 was higher (p < 0.05) in normal pregnancy. This response was positively correlated with CX3CR1 expression. Blockade of CX3CR1 canceled the secretory response to LPS in all groups.ConclusionsChA-complicated pregnancy up-regulates CX3CR1 in HAEC cultured in vitro with simultaneous increase in CX3CL1 production. Hypoxia-resistant production of CX3CL1 may be responsible for ChA-related complications of pregnancy and labor.

Randomized Controlled Trial Evaluating Dialkylcarbamoyl Chloride Impregnated Dressings for the Prevention of Surgical Site Infections in Adult Women Undergoing Cesarean Section

Abstract Background: Surgical site infections (SSI) occur in 1.8%–9.2% of women undergoing cesarean section (CS) and lead to greater morbidity rates and increased treatment costs. The aim of the study was to evaluate the efficacy and cost-effectiveness of dialkylcarbamoyl chloride (DACC) impregnated dressings to prevent SSI in women subject to CS. Methods: Randomized, controlled trial was conducted at the Mazovian Bródno Hospital, a tertiary care center performing approximately 1300 deliveries per year, between June 2014 and April 2015. Patients were randomly allocated to receive either DACC impregnated dressing or standard surgical dressing (SSD) following skin closure. In order to analyze cost-effectiveness of the selected dressings in the group of patients who developed SSI, the costs of ambulatory visits, additional hospitalization, nursing care, and systemic antibiotic therapy were assessed. Independent risk factors for SSI were determined by multivariable logistic regression. Results: Five hundred and forty-three women undergoing elective or emergency CS were enrolled. The SSI rates in the DACC and SSD groups were 1.8% and 5.2%, respectively (p = 0.04). The total cost of SSI prophylaxis and treatment was greater in the control group as compared with the study group (5775 EUR vs. 1065 EUR, respectively). Independent risk factors for SSI included higher pre-pregnancy body mass index (adjusted odds ratio [aOR] = 1.08; [95% confidence interval [CI]: 1.0–1.2]; p < 0.05), smoking in pregnancy (aOR = 5.34; [95% CI: 1.6–15.4]; p < 0.01), and SSD application (aOR = 2.94; [95% CI: 1.1–9.3]; p < 0.05). Conclusion: The study confirmed the efficacy and cost-effectiveness of DACC impregnated dressings in SSI prevention among women undergoing CS.

Value of ultrasonography in the diagnosis of polycystic ovary syndrome – literature review

Polycystic ovary syndrome is a multi-factorial disease. Its etiopathogenesis has not been elucidated in detail. It is the most common endocrine disorder in women of child-bearing age. This disease entity is primarily characterized by disrupted ovulation and hyperandrogenism, but the clinical picture can be diversified and symptom intensity can vary. Currently, the sonographic assessment of ovaries is one of the obligatory criteria for the diagnosis of PCOS according to the Rotterdam consensus (2003) and Androgen Excess & PCOS Society (2006). This criterion is determined by the presence of ≥12 follicles within the ovary with a diameter of 2–9 mm and/or ovarian volume ≥10 cm3. Such an ultrasound image in one gonad only is sufficient to define polycystic ovaries. The coexistence of polycystic ovaries with polycystic ovary syndrome is confirmed in over 90% of cases irrespective of ethnic factors or race. However, because of the commonness of ultrasound features of polycystic ovaries in healthy women, the inclusion of this sign to the diagnostic criteria of polycystic ovary syndrome is still questioned. The development of new technologies has an undoubted influence on the percentage of diagnosed polycystic ovaries. This process has caused an increase in the percentage of polycystic ovary diagnoses since the Rotterdam criteria were published. It is therefore needed to prepare new commonly accepted diagnostic norms concerning the number of ovarian follicles and the standardization of the technique in which they are counted. The assessment of anti-Müllerian hormone levels as an equivalent of ultrasound features of polycystic ovaries is a promising method. However, analytic methods have to be standardized in order to establish commonly accepted diagnostic norms.

Impact of pre-gestational and gestational diabetes mellitus on the expression of glucose transporters GLUT-1, GLUT-4 and GLUT-9 in human term placenta

PurposeVarious studies in placental tissue suggest that diabetes mellitus alters the expression of glucose transporter (GLUT) proteins, with insulin therapy being a possible modulatory factor. The aim of the present study was quantitative evaluation of the expression of glucose transporters (GLUT-1, GLUT-4, GLUT-9) in the placenta of women in both, uncomplicated and diabetic pregnancy. Additionally, the effect of insulin therapy on the expression of selected glucose transporter isoforms was analyzed.MethodsTerm placental samples were obtained from healthy control (n = 25) and diabetic pregnancies, including diet-controlled gestational diabetes mellitus (GDMG1) (n = 16), insulin-controlled gestational diabetes mellitus (GDMG2) (n = 6), and pre-gestational diabetes mellitus (PGDM) (n = 6). Computer-assisted quantitative morphometry of stained placental sections was performed to determine the expression of selected glucose transporter proteins.ResultsMorphometric analysis revealed a significant increase in the expression of GLUT-4 and GLUT-9 in insulin-dependent diabetic women (GDMG2 + PGDM) as compared to both, control and GDMG1 groups (p < .05). Significantly increased GLUT-1 expression was observed only in placental specimens from patients with PGDM (p < .05). No statistically significant differences in GLUT expression were found between GDMG1 patients and healthy controls.ConclusionsThe results of the study confirmed the presence of GLUT-1, GLUT-4 and GLUT-9 proteins in the trophoblast from both, uncomplicated and diabetic pregnancies. In addition, insulin therapy may increase placental expression of GLUT-4 and GLUT-9, and partially GLUT-1, in women with GDMG2/PGDM.

Expression of Glucose Transporter Proteins in Human Diabetic Placenta

Gestational diabetes mellitus and pregestational diabetes mellitus constitute carbohydrate metabolism disorders, which, if not diagnosed and adequately treated, lead to serious and often life-threatening pregnancy complications. According to a recently formulated hypothesis, some diabetes-related complications, such as fetal macrosomia, may be the result of disturbances in the transplacental transport of nutrients-in particular, excessive maternal-fetal glucose transfer. Throughout pregnancy, glucose flux across the placenta is mediated by the group of facilitative glucose transporters (GLUT), the expression of which in different placental compartments is the precondition for effective glucose uptake from maternal blood and its subsequent transfer to the fetal circulation. In diabetes-complicated pregnancies, the location, expression and activity of glucose transporters are modified to an extent that results in alterations in the maternal-fetal glucose exchange, potentially leading to an excessive supply of energy substrates to the fetus. This paper reviews the literature on the expression and activity of glucose transporter proteins-GLUT-1, GLUT-3, GLUT-4, GLUT-8, GLUT-9 and GLUT-12-in the human placenta, with a special focus on diabetes-complicated pregnancy. The characteristics of transporters in conditions of maternal normoglycemia and modifications occurring in the diabetic placenta are summarized, and the factors responsible for the regulation of the expression of selected isoforms are described. Finally, the impact of alterations in the placental expression of the aforementioned members of the GLUT family on intrauterine fetal development in pregnancies complicated by diabetes mellitus is discussed.

Spontaneous rupture of the pregnant uterus following salpingectomy: a literature review.

Background: Spontaneous rupture of the uterus is a life-threatening obstetric complication in rare cases associated with previously performed salpingectomy. Aim: This paper presents an analysis of uterine rupture cases during pregnancy in patients after surgical removal of the fallopian tubes. Methods: The English and Polish language literature was reviewed for studies published between January 1, 1980 and September 30, 2014 to identify articles that described rupture of the uterus in women with a previous history of salpingectomy. Results: Thirteen case reports and case series studies in 18 women were identified. 33% of cases of uterine rupture following salpingectomy occurred during intrauterine pregnancy, whereas the rest was associated with interstitial ectopic pregnancy. Laparoscopic salpingectomy more often resulted in rupture of the uterus during non-ectopic pregnancy as compared to laparotomy (4 vs. 2 cases, respectively). When interstitial pregnancies were excluded, uterine rupture was a cause of fetal death in 67% of reported gestations. There were no cases of maternal mortality. Conservative treatment was the preferred management option, and total hysterectomy was performed in only 2 patients. Conclusion: Particular attention should be paid to patients with a previous history of salpingectomy due to the risk of uterine rupture throughout the entire pregnancy.