Ultra-high-speed liquid chromatography combined with mass spectrometry detection analytical methods for the determination of nitrosamine drug substance-related impurities.

In the present study, five simple, feasible, and sensitive Ultra-high-speed liquid chromatography combined with mass spectrometry detection methods, using electrospray ionization are proposed. These methods were developed and validated for the determination of four different nitrosamine drug substance-related impurities-N-nitrosoacebutolol, N-nitrosobisoprolol, N-nitrosometoprolol, and N-nitrososotalol-in five beta blockers active pharmaceutical ingredients-acebutolol HCl, bisoprolol fumarate, metoprolol tartrate, metoprolol succinate, and sotalol HCl. The proposed methods were validated as per regulatory guidelines. Acquity HSS T3 (3.0 × 100 mm, 1.8 µm) column and formic acid 0.1% in water combined with methanol or acetonitrile were used for chromatographic separation in all methods. The limit of detection and the limit of quantification were found to be in the range of 0.02-1.2 and 2-20 parts per billion, respectively. The accuracy and precision of the five methods have been demonstrated in the working range of each one, giving values of recovery within the range of 64.1%-113.3%, and the regression coefficients (R) were found to be in the range of 0.9978-0.9999. These methods could be used for controlling nitrosamine drug substance-related impurities content for beta blockers drug substances batches manufactured at Moehs group.

Risk factors for relapse or recurrence in women with bipolar disorder and recurrent major depressive disorder in the perinatal period: a systematic review.

It is well known that the perinatal period supposes a considerable risk of relapse for women with bipolar disorder (BD) and recurrent major depressive disorder (rMDD), with the consequences that this entails. Therefore, the authors sought to provide a critical appraisal of the evidence related to specific risk factors for this population with the aim of improving the prevention of relapses during pregnancy and postpartum. The authors conducted a systematic review assessing 18 original studies that provided data on risk factors for relapse or recurrence of BD and/or rMDD in the perinatal period (pregnancy and postpartum). Recurrences of BD and rMDD are more frequent in the postpartum period than in pregnancy, with the first 4-6 weeks postpartum being especially complicated. In addition, women with BD type I are at higher risk than those with BD type II and rMDD, and the most frequent presentation of perinatal episodes of both disorders is a major depressive episode. Other risk factors consistently repeated were early age of onset of illnesses, severity criteria, primiparity, abrupt discontinuation of treatment, and personal or family history of perinatal affective episodes. This review shows that there are common and different risk factors according to the type of disorder and to perinatal timing (pregnancy or postpartum) that should be known for an adequate prevention of relapses.

Concentrations of Selected Adipocytokines in the Blood Plasma in Proximal Suspensory Desmopathy of Horses, with a Focus on Their Physical Activity-A Pilot Study.

Chronic tendon and ligament diseases are commonly encountered in both athletic humans and animals, especially horses. Distal limb diseases, including suspensory ligament (SL) pathology due to anatomical, histological, and biomechanical properties, can be considered a model for tendon and ligament pathologies in humans. The appropriate selection of therapy is often crucial in optimising the healing process. One decisive factor influencing the possibility of returning to pre-disease training levels appears to be the utilisation of physical activity, including controlled movement, during the rehabilitation process. In the pathogenesis of musculoskeletal diseases and rehabilitation, adipocytokines play diverse roles. However, it is unclear what significance they hold in horses and in specific disease entities as well as the consequences of their mutual interactions. Recent studies indicate that in the pathogenesis of diseases with varied aetiologies in humans, their value varies at different stages, resulting in a diverse response to treatment. The results of this study demonstrate lower resistin concentrations in the venous blood plasma of horses with proximal suspensory desmopathy (PSD), while higher levels were observed in regularly trained and paddocked animals. The horses investigated in this study showed higher concentrations of resistin and IL-8, particularly in paddocked horses as well as in the working group of horses. The results suggest that these concentrations, including resistin in blood plasma, may be clinically significant. This attempt to explore the aetiopathogenesis of the processes occurring in the area of the proximal attachment of the suspensory ligament may optimise the procedures for the treatment and rehabilitation of horses.

Treating postpartum affective and/or anxiety disorders in a mother-baby day hospital: preliminary results.

INTRODUCTION: Women experiencing perinatal mental-health illness have unique needs. The present study analyzes preliminary data about the effectiveness of MBDH in treating postpartum women with affective and anxiety disorders. METHODS: We analysed 33 mothers with affective and/or anxiety disorders treated at the MBDH with their babies between March 2018 and December 2019. All women were assessed at admission, discharge and three months after discharge. Outcomes included symptoms of depression (EPDS) and anxiety (STAI-S), mother-infant bonding (PBQ) and functional impairment (HoNOs). We also assessed the clinical significance of changes in patients' scores on these scales and patient satisfaction. RESULTS: At discharge, no patients still met the full criteria for the main diagnosis. Between admission and discharge, symptoms of depression and anxiety, mother-infant bonding, functional impairment and autonomy in caring for babies improved significantly. These gains were maintained at three months follow-up. Patient satisfaction was high. CONCLUSIONS: These preliminary results suggest that multidisciplinary intervention for postpartum women with affective or anxiety disorders at the MBDH improves maternal psychopathology, mother-infant bonding and mothers' ability to care for their babies. MBDHs are a promising approach for delivering specialised perinatal mental-health care for mother-baby dyads.

Mother-baby day hospital (MBDH) could be an adequate device for women with perinatal mental disordersMultidisciplinary intervention is effective for treating postpartum anxiety and depressive disorders.Interventions at MBHD improve mother-infant bonding and mothers' ability to care for their babies.Further research is needed to assess the effectiveness at long term not only on maternal health also on child neurodevelopment.

The Importance of Immunological Disorders in the Pathogenesis of Lichen Sclerosus in Pediatric Patients: A Systematic Review.

Lichen sclerosus (LS) is defined as a chronic mucocutaneous inflammatory disease with a localization predominantly to the anus and genitals (vulvar sclerosus (VLS)). Pediatric lichen sclerosus (LS) is a chronic inflammatory skin condition with predilection for the anogenital area that if untreated can lead to scarring. Vulvar LS is characterized by two peaks in incidence: it occurs in prepubertal girls and in postmenopausal women. To date, several mechanisms and risk factors have been proposed in the pathogenesis of pediatric vulvar LS; however, the etiology of this condition is still not fully understood and constitutes a challenge for scientists and clinicians. The presented research aimed to systematically review the existing literature on the pathogenesis of pediatric LS and to identify possible underlying autoimmune mechanisms and molecular networks. The clinical presentation of pediatric lichen sclerosus and available treatment modalities are also presented to acquaint a broader audience with this underdiagnosed and undertreated condition. As a result of our review, we discuss several potential mechanisms, molecules, and pathways that have been recognized in this disease. The purpose of our review was also to summarize what we can induce in further studies, which will ultimately help to identify the mechanism responsible for the disease and aid in the development of new, more effective treatment strategies for diagnosis and treatment by clinicians and researchers.

Hybrid simulation of pediatric gynecologic examination: a mix-methods study of learners' attitudes and factors affecting learning.

BACKGROUND: Children and adolescents with reproductive health issues may require immediate or emergency care, however in many countries availability of the pediatric and adolescent gynecology (PAG) service is low. That being said, teaching PAG examination to OBGYN, pediatrics and emergency medicine residents seems reasonable, and cannot be underestimated. In order to provide residents with opportunity to learn PAG examination, a high-fidelity hybrid simulation workshop was implemented in our institution. METHODS: The study aimed to investigate learners' attitudes towards the high-fidelity simulation (HFS) hybrid model as compared with task trainer-SP (simulated patient)-voice model in the HFS environment and the factors connected to learners' attitudes towards the hybrid model that could influence learning in high-fidelity simulation (HFS). The concept of attitude was used as the theoretical framework and the mixed method approach to study design was utilized with simultaneous collection of quantitative (original questionnaires) and qualitative data (semi-structured interviews). RESULTS: Residents valued the HFS hybrid model higher over task trainer-SP-voice model in regards to all three attitude components: cognitive (95%), affective (87.5%) and behavioral (83.7%). Analysis of qualitative data revealed six themes important to learners and informing learning of PAG examination in HFS. Further analysis of the themes allowed to develop a conceptual model, in which six factors connected to attitude components influenced learning. These factors were: task difficulty, attention, emotional realism of the simulation, patient's emotions, physical realism of the simulation, and technical issues. CONCLUSIONS: Participants of our study appreciated learning experience with the HFS hybrid model more, based on attitude questionnaire. Moreover, findings revealed that multiple, various factors connected to attitude may influence learning of PAG examination in HFS with hybrid model, and we propose a conceptual model illustrating relationships between those factors.

Changes of post-operative peripheral blood dendritic cells in patients undergoing laparoscopy.

INTRODUCTION: Surgical intervention affects local and systemic immune responses, especially in obese individuals. Many studies have attempted to evaluate immunological response to surgical trauma. Surgery changes the quantity and phenotype of circulating blood dendritic cells (DCs), including a decrease of total DCs post-operatively. The study aimed to evaluate the percentage and changes of myeloid, lymphoid DCs, and myeloid to lymphoid DCs ratio in obese and normal weight patients undergoing laparoscopy. MATERIAL AND METHODS: The study enrolled asymptomatic patients with gallstones, who underwent laparoscopic cholecystectomy. Blood samples were obtained before the surgery as well as 24 and 48 hours after the surgery. Cells were collected using a FACSCalibur flow cytometry, and phenotypes were analyzed with CellQuest software. RESULTS: No statistically significant differences were observed between obese and normal-weighted patients in all studied time periods, except for the myeloid to lymphoid DCs ratio assessed at 48-post-operative hour. The myeloid DCs percentage increased significantly in the post-operative period within both studied groups. The percentage of lymphoid DCs increased significantly in obese patients in all studied time periods. CONCLUSIONS: Laparoscopy induces immunomodulation, such as changes of myeloid and lymphoid dendritic cells, especially in obese patients. We describe new findings, in which minimally invasive surgical trauma promotes the increase of percentage of circulating DCs in the early post-operative period.

CD44, TGM2 and EpCAM as novel plasma markers in endometrial cancer diagnosis.

BACKGROUND: Endometrial cancer (EC) is the most common malignancy of the female reproductive tract. Despite years of research, the accurate screening strategy is still not available in this disease and it is usually diagnosed only after the clinical signs are present. The recent technological advances in analytical methodologies enabled detection of multiple molecules in one, small sample of biological materials. Such approach was undertaken in the presented study. METHODS: Concentrations of aldehyde dehydrogenase 1 family, member A1 (ALDH1A1), carbonic anhydrase IX (CA9), CD44, epithelial cell adhesion molecule (EpCAM), hepsin, kallikrein-6, mesothelin, midkine, neural cell adhesion molecule L1 (L1CAM), and transglutaminase 2 (TGM2) were measured using MAGPIX®System in plasma samples of 45 EC, 20 healthy controls and 11 patients with endometriosis. RESULTS: Significantly increased concentration in EC as compared to healthy controls were found in case of CD44 (p <  0.001), EpCAM (p = 0.033) and TGM2 (p <  0.001). EpCAM and mesothelin concentrations differed based on FIGO stages. Regression analysis revealed marker panels with high accuracy in detection of EC. The highest AUC 0.937 was attributed to the 3-marker panel of CD44/TGM2/EpCAM (84% sensitivity, 100% specificity), FIGO IA samples were discriminated from more advanced stages of EC with the mesothelin/grade 1 model featuring AUC of 0.911 (95.24% sensitivity, 78.26% specificity). CONCLUSIONS: Novel plasma biomarkers presenting good accuracy in diagnosing EC were found with TGM2 reported for the first time as plasma marker. It was also revealed that endometriosis may share similarities in the pattern of markers alterations characteristic for EC.

Interactions between microRNA-200 family and Sestrin proteins in endometrial cancer cell lines and their significance to anoikis.

In the present study, we intend to determine whether Sestrin proteins 1, 2, and 3 (SESN1-3) are targets of microRNA-200 family (miR-200) in endometrial cancer (EC) Ishikawa, AN3CA, KLE, and RL 95-2 cell lines and to investigate how these potential interactions influence anoikis resistance of EC cell lines. The luciferase reporter assay, qRT-PCR, and western blotting assays were used to verify whether SESN1-3 are direct targets of miR-200. Moreover, the anoikis assay and transient transfections of miR-200 mimics or inhibitors into EC cell lines were performed to evaluate the modulatory role of miR-200 and SESN proteins on anoikis resistance. We demonstrated that SESN2 protein is a direct target of mir-141 in KLE and RL-95-2 EC cell lines and the functional interaction of miR-141 and SESN2 protein has a downstream effect on anoikis resistance and SESN2 expression level in Ishikawa and AN3CA cell lines. Moreover, we have shown that SESN3 protein is a direct target of miR-200b, miR-200c, and miR-429 in Ishikawa, AN3CA, and KLE cell lines. Our results show that manipulation of miR-200b, miR-200c, and miR-429 expression patterns also has an influence on anoikis resistance in EC cell lines. In conclusion, we identified new interactions between miR-200 and the oxidative stress response SESN proteins that affect anoikis resistance in human EC cells.

Mother-infant bonding in the postpartum period: assessment of the impact of pre-delivery factors in a clinical sample.

This study aims to examine the extent to which a variety of pre-delivery factors (demographic, reproductive, psychological, psychiatric, and psychopathological) predict disturbances in mother-infant bonding (MIB) in the postpartum period. Two hundred fifty-one pregnant women enrolled at a public perinatal psychiatric service were assessed between the first and second trimester of pregnancy and at 6-7 weeks after delivery. During pregnancy, the psychological risk factors were assessed with the Vulnerable Personality Style Questionnaire, the Marital Adjustment Scale, the Early Trauma Inventory, and the General Health Questionnaire. To detect psychopathology, the Edinburgh Postnatal Depression Scale and the State-Trait Anxiety Inventory were used. At the postpartum evaluation, MIB was measured by the Postpartum Bonding Questionnaire. The results of the final regression model showed that emotional abuse in childhood, family psychiatric history, previous psychiatric hospitalization, and anxiety during pregnancy were significant predictors of MIB disturbances in postpartum, explaining 10.7% of the variance. The evaluation of women's risk factors in pregnancy is important in order to prevent MIB disturbances and thus to ensure the welfare of mothers and their babies.

Prevalence of Small-Airway Dysfunction among COPD Patients with Different GOLD Stages and Its Role in the Impact of Disease.

BACKGROUND: In chronic obstructive pulmonary disease (COPD) patients, small-airway dysfunction (SAD) is considered a functional hallmark of disease. However, the exact role of SAD in the clinical presentation of COPD is not yet completely understood; moreover, it is not known whether SAD may have a relationship with the impact of disease. OBJECTIVES: To evaluate the prevalence of SAD among COPD patients categorized by the old and the new GOLD classification and to ascertain whether there is a relationship between SAD and impact of disease measured by the COPD Assessment Test (CAT) questionnaire. METHODS: We prospectively enrolled COPD outpatients from the University Hospital of Parma. Using the impulse oscillometry system (IOS), we assessed the fall in resistance from 5 to 20 Hz (R5-R20), reactance at 5 Hz (X5), and resonant frequency (FRes) as markers of peripheral airway dysfunction. According to R5-R20 ≥0.07 or <0.07, the cohort was also categorized in patients with and without SAD, respectively. RESULTS: We studied 202 patients. In both GOLD classifications, a progressive increasing distribution of R5-R20 and FRes was reported with a decreasing of X5. Moreover, there was a significant correlation between R5-R20 and CAT (r = 0.527, p < 0.001). Finally, the presence of SAD (OR 11.96; 95% CI 4.53-31.58; p < 0.001) and use of ICS + LABA + LAMA (OR 5.31; 95% CI 1.88-15.02; p = 0.002) were independent predictors of higher impact (CAT score ≥10). CONCLUSION: In COPD patients, the presence of SAD, as assessed by IOS, progressively increases with GOLD classifications and it is closely related to the high impact of disease on health status.

What Do We Know About Education in Colorectal Cancer Prevention?-Survey Among 1130 Medical Students.

Colorectal cancer (CRC) is a global public health problem. The degree of knowledge on CRC among medical students, future physicians, brings essential implications for their patients. Therefore, to acquire information about students' knowledge on CRC, a questionnaire survey was conducted in Medical University of Lublin, Poland, with a representative sample of 1130 medical students (basic vs. clinical 552 vs. 578; male vs. female 442 vs. 688). The questionnaire was anonymous and designed in a four-section scheme (CRC risk factors, CRC prevention, CRC symptoms, CRC screening methods). There was a significant difference in the percentage of correct answers between basic and clinical level groups (P = 0.034). In general, clinical students had higher scores for their knowledge regarding CRC. Gender impacted the students' CRC knowledge to a lesser degree; however, if the difference was revealed, female students were more aware about CRC issues (P = 0.045). We found several important deficits in students' knowledge on CRC. These findings should motivate the oncology education coordinator as well as our teachers to introduce innovations in education methods and training environments to enable students to gain necessary knowledge and acquire the skills and competencies that would help them to function as physicians.

In vitro and in vivo activity of miR-92a-Locked Nucleic Acid (LNA)-Inhibitor against endometrial cancer.

BACKGROUND: Endometrial cancer is the most common cancer of the female reproductive tract. Based on our previous studies we speculated that miR-92a exhibited pro-oncogenic properties in endometrial cancer, and therefore its inhibition could be used as a therapeutic measure in this disease. Therefore in the present study we aimed to investigate both in vitro and in vivo if inhibition of miR-92a in endometrial cancer would limit cancer cells proliferation. METHODS: miR-92a expression was evaluated in four endometrial cancer cell lines using qPCR. Inhibition of miR-92a activity was obtained in endometrial cancer cell lines by a transient transfection of a custom designed Locked Nucleic Acid (LNA)-Inhibitor, developed to work both in vitro and in vivo. In vitro proliferation studies were performed using xCELLigence RTCA DP system. In vivo experiment was performed in Cby.Cg-Foxn1 < nu>/cmdb mice bearing endometrial cancer xenografts, which were intraperitoneally injected with nine dosages of 25 mg/kg of miR-205-LNA-inhibitor. RESULTS: qPCR revealed increased expression of miR-92a in HEC-1-B, Ishikawa and AN3CA cells. LNA-i-miR-92a inhibited endometrial cancer growth in vitro. It was also demonstrated that systemic administration of LNA-i-miR-92a was feasible and exerted inhibitory effect on endometrial cancer xenograft growth in vivo with only mild toxic effects in treated animals, however the effect was observed until 12th experimental day and the last three dosages did not maintain the attenuating effect with the acceleration of tumor growth observed at the end and after cessation of the intraperitoneal therapy. CONCLUSIONS: Taken together, these results indicate that intraperitoneal delivery of miR-92a-LNA-modified-inhibitor is feasible, devoid of significant toxicity and moderately inhibits endometrial cancer growth in vivo, and therefore warrants further studies investigating other routes of inhibitor delivery possibly in other animal models.

Assessment of psychometric properties of the Postpartum Bonding Questionnaire (PBQ) in Spanish mothers.

The Postpartum Bonding Questionnaire (PBQ) was developed to assess mother-infant bonding disturbances in the postpartum period. The aim of this study was to examine the psychometric properties of the Spanish version of the PBQ in a sample of Spanish postpartum women. Eight hundred forty mothers were recruited in the postpartum visit (4-6 weeks after delivery): 513 from a gynecology unit (forming the general population sample) and 327 mothers from a perinatal psychiatry program (forming the clinical sample). All women were assessed by means of the Edinburgh Postnatal Depression Scale (EPDS) and the PBQ. Neither the original four-factor structure nor alternative structures (Reck et al. 2006; Wittkowski et al. 2010) were replicated by the confirmatory factor analyses. An exploratory factor analysis showed a four-factor solution. The Schmid-Leiman transformation found a general factor that accounted for 61% of the variance of the PBQ. Bonding impairment showed higher associations with depressive symptomatology in both samples. The Spanish version of the PBQ showed adequate psychometric properties for use with clinical and general populations of Spanish postpartum women. The results suggest that the PBQ could be summarized by a general factor and confirm the utility of the use of the total score for detecting bonding impairment.

Clinically relevant anatomical parameters of the replaced right hepatic artery (RRHA).

PURPOSE: Vascular anatomy of the liver is subjected to many variations. The most common hepatic artery (HA) replacement is the right hepatic artery (RRHA). Variations of the HA are particularly important consideration when choosing the best surgical procedure or if radiological abdominal intervention is required. In this study, we evaluated the anatomical details of the RRHA origin. METHODS: Retrospective investigation of clinical data from 1569 patients who underwent an abdominal MDCT was performed. The anatomy of RRHA origin was described based on four parameters measured: D--the distance between SMA origin and the RRHA origin, L--the lumen at the place of origin, AH--the origin angle from the SMA in horizontal plane, and AV--the origin angle from the SMA in vertical plane. RESULTS: RRHA arising from SMA was detected in 10.13 % of cases (159/1569) and its anatomy was subjected to variations. Mean (±SD) of parameters D, L, AH and AV was 27.34 mm ± 6.83, 3.29 mm ± 1.17, 97.27º ± 26.69 and 89.73º ± 20.81, respectively. Values of parameters D and L were significantly higher in males compared to females. CONCLUSION: Although radiologists are not always aware of the clinical significance of the RRHA origin, the evaluation of its anatomy is thought to help reduce the risk of inadvertent vascular injury, especially in pancreatoduodenectomy. Detection and evaluation of the RRHA does not necessarily require angio-CT examination. Our study demonstrated that the MDCT, the standard imaging modality for diagnosing the abdominal symptoms, is sufficient to provide the knowledge of the HA abnormalities.

Assessment of the correlation between gender, age, body mass index and the severity of postoperative pain, nausea and vomiting in patients undergoing laparoscopic cholecystectomy.

UNLABELLED: Obesity has become one of the leading epidemic diseases in the world. Obesity is acknowledged as a risk factor for postoperative pain, nausea and vomiting (PONV), especially in obese patients undergoing laparoscopic procedures. AIM: The aim of the study was to evaluate the correlation between gender, age, BMI values and the severity of postoperative pain and PONV in patients undergoing laparoscopic cholecystectomy. MATERIALS AND METHODS: The study enrolled 244 patients with cholelithiasis who underwent scheduled laparoscopic cholecystectomy. Patients were assigned to 3 groups according to BMI values. Patients were assessed for the presence of PONV and severity of pain in the 6(th) and 24(th) hour after surgery. RESULTS: BPONV was reported more frequently in female than in male patients in the 1(st) time period. In the first time point the postoperative pain was significantly higher in group O in comparison to group H and N. Postoperative pain significantly decreased in the 24(th) postoperative hour in comparison to its level assessed in the 6th postoperative hour in all three groups. CONCLUSIONS: Female gender is a risk factor of PONV, so women should receive antiemetic prophylactic. There is a positive relationship between BMI and postoperative pain, so obese patients undergoing laparoscopic procedures should receive prophylactic application of analgesic drugs.

[Medical simulation as a professional tool which affect the safety of the patient used in the learning process]. / Symulacja medyczna jako profesjonalne narzedzie wplywajace na bezpieczenstwo pacjenta wykorzystywane w procesie nauczania.

Simulation is the methodology of teaching, learning process using educational equipment from simple simulators, designed to study individual tasks, through advanced mannequins called human patient stimulators faithfully mimic the human and its parameters. The medical simulation's main task is education and improvement patients' safety. Advanced human simulators can realistically cough, vomit with artificial chyme and bleed with artificial blood causing a real stress of medical personnel and the need for immediate action. Medical simulation gives the opportunity to prepare medical personnel to the profession more effectively, in less time than traditional education and also clearly affects the patients' safety.

Does thermodynamic stability of peritoneal collagen change during laparoscopic cholecystectomies? A differential scanning calorimetry (DSC) study.

BACKGROUND: Carbon dioxide pneumoperitoneum used during laparoscopic surgeries alters the integrity of the peritoneum and results in denudation of the basal lamina that might cause altered immune response, inhibited fibrinolysis, hypoxia, and acidosis. The changes in the structure of pneumoperitoneum were described as bulging of mesothelial cells, irregular cell junction's cell membrane degradation, and mesodermal edema. As denaturation of peritoneal proteins reflects overall condition of its structure and interactions with the surrounding molecules, the physical status of collagen was assessed on the basis of parameters of thermal denaturation measured by DSC method. METHODS: Twenty-four female patients operated on due to cholelithiasis were enrolled in this study. Laparoscopic cholecystectomy was performed using standard four-trocar technique, and standard values of insufflated carbon dioxide pneumoperitoneum were used. After trocar placement, the first collection of peritoneal sample (sample A) was performed. The second peritoneal sample (sample B) was collected after the removal of gall bladder. Differential scanning calorimetry (Q200 calorimeter, TA Instruments) was performed on samples defrosted at room temperature. RESULTS: In all samples of peritoneum, a nonreversible endothermal process recognized as denaturation was observed. Sample B obtained at the end of surgery did not differ from sample A obtained at the beginning in terms of all parameters under study. Temperature of denaturation in A and B was correlated only marginally, but enthalpy and specific heat were significantly correlated. The analysis of data from DSC measurements did not reveal differences in physical stability of collagen in peritoneal samples obtained at the beginning and at the end of surgery. Significant negative correlations between duration of CO2 pneumoperitoneum and enthalpy of denaturation in sample B were found. CONCLUSIONS: Differences in enthalpy of denaturation may reflect a quantitative relation between amount of native collagen molecules in the sample and other, non-collagenous components or impaired collagen.

Diagnostic and prognostic significance of miRNA signatures in tissues and plasma of endometrioid endometrial carcinoma patients.

The aim of our study was to define tissue and plasma miRNA signatures, which could potentially serve as diagnostic and prognostic markers in endometrioid endometrial cancer (EEC) and to investigate miRNA profiles in regard to clinicopathological characteristics. Tissue and plasma samples were collected from 122 women (77 EEC and 45 controls). Expression profiling of 866 human miRNAs and 89 human viral miRNAs was performed in 24 samples and was followed by qPCR validation in 104 patients. Expression of 16 miRNAs was analyzed in 48 plasma samples. Microarray study revealed regulation of 21 miRNAs in EEC tissues comparing to normal endometrium. Altered expression of 17 miRNAs was confirmed by qPCR performed in 104 tissue samples. Seven miRNAs were upregulated and two were downregulated in EEC plasma samples. Expression of a number of miRNAs was associated with International Federation of Gynecology and Obstetrics stage, grade, relapse and nodal metastases. Two miRNA signatures: miR-92a/miR-410 and miR-92a/miR-205/miR-410 classified tumor tissues with higher accuracy in comparison to single miRNAs (AUC: 0.977, 95% CI: 0.927-0.996 and 0.984, 95% CI: 0.938-0.999, respectively). miRNA signature composed of miR-205 and miR-200a predicted relapse with AUC of 0.854 (95% CI: 0.691-0.951). Tissue miRNA signatures were independent prognostic markers of overall (miR-1228/miR-200c/miR-429, HR: 2.98) and progression-free survival (miR-1228/miR-429, HR: 2.453). Plasma miRNA signatures: miR-9/miR-1228 and miR-9/miR-92a, classified EEC plasma samples with high accuracy yielding AUCs of 0.909 (95% CI: 0.789-973) and 0.913 (95% CI: 0.794-0.976), respectively. We conclude that miRNA signatures hold a great promise to become noninvasive biomarkers for early EEC detection and prognosis.