Hamman's Syndrome after Vaginal Delivery: A Case of Postpartum Spontaneous Pneumomediastinum with Subcutaneous Emphysema and Review of the Literature.

Hamman's syndrome is a rare condition that mostly affects young males, often with a predisposition to asthma. It includes the presence of free air in the mediastinum and subcutaneous emphysema with no other underlying cause such as trauma, infection, or administration of any sort of mask support with hyperpressure. It occurs spontaneously and often in association with a prolonged Valsalva maneuver. This might explain why there are some cases of Hamman's syndrome among young females giving birth. Here, we present a case report of a 24-year-old non-smoker primigravida with Hamman's syndrome. She presented with symptoms a few hours after an uncomplicated vaginal delivery at 40 + 1 weeks of pregnancy where the active phase of labor lasted for three hours with normal progress. The second stage lasted for 30 min, with no signs of distress on CTG. The symptoms (pain in the right ear, swelling and pain in the neck, chest tightness, shortness of breath, dysphagia, odynophagia, and pain in the upper thorax on the right side) and objective findings as subcutaneous crepitations in the neck, parasternal region, right axillary fossa, clavicle and over the chest resolved spontaneously after a few days of observation and conservative management. We also give a systemic review of reported cases since 2000 to provide an overview of the pathomechanism, symptoms, diagnostics, treatment, and management of this condition. Hamman's syndrome is a rare, usually benign, but potentially serious complication that can occur during the second stage of labor. Diagnostics include inquiring about typical symptoms, clinical examination, and chest x-ray or CT scan. Treatment is usually conservative with oxygen, bronchodilators, and pain relief. The recurrence rate is low and there is no contraindication to vaginal delivery in future pregnancies. However, it is suggested that physicians and midwives be cautious and consider a low threshold for instrumental delivery or cesarean section to avoid excessive Valsalva maneuvers.

Desmoid Fibromatosis of the Anterior Abdominal Wall in Pregnancy: A Case Report and Review of the Literature.

A desmoid tumor (DT) is a rare benign neoplasm arising from muscle aponeurosis, associated mostly with trauma or pregnancy. DT has an infiltrative and locally aggressive growth pattern and usually does not metastasize. However, it has a high recurrence and complication rate. When it occurs in pregnancy, the pregnancy and delivery is taken as an individual case for optimal management by physicians and midwifes, who need to be cautious in finding the optimal delivery mode for the patient, which depends on the tumor size, location, behavior, and past history. The authors report a case of 29-year-old pregnant woman who previously underwent systemic oncological treatment for a large abdominal wall desmoid tumor and became pregnant afterwards. The history of DT presented a follow-up and delivery challenge. Observational management was chosen with an elective cesarean section at week 38 + 4 of pregnancy with uncomplicated postpartum follow-up. The authors detail the clinical management and chosen therapeutic approach; chemotherapy can be a choice in the treatment options for patients with DTs, although the majority of DTs are treated surgically with subsequent mesh plastic. Moreover, the authors provide a systematic review of the literature focused on the treatment management of DTs in pregnant women during pregnancy and the postpartum period, as pregnancy-associated desmoid tumors are a specific condition, where the optimal management is not well established, despite some guidelines for non-pregnant patients.

Cognitive Functions, Depressive and Anxiety Symptoms After One Year of CPAP Treatment in Obstructive Sleep Apnea.

Objective: The study worked with depressive symptoms, anxiety score and cognitive functions in obstructive sleep apnea (OSA) patients treated with CPAP. Methods: Eighty-one subjects with OSA and without psychiatric comorbidity were treated with CPAP for one year and completed the following scales and cognitive tests: Trail Making Test, Verbal Fluency Test, d2 Test, Beck Depression Inventory-II and Beck Anxiety Inventory. MINI ruled out psychiatric disorder. At the two months check-up, subjects were re-evaluated for depressive and anxiety symptoms, and after one year of CPAP treatment, subjects repeated cognitive tests and scales. Data about therapy adherence and effectiveness were obtained from the patient's CPAP machines. Results: The study was completed by 59 CPAP adherent patients and eight non-adherent patients. CPAP therapy effectiveness was verified in all patients by decreasing the apnea-hypopnoea index below 5 and/or 10% of baseline values. The adherent patients significantly improved depressive and anxiety symptoms. There was also an improvement in overall performance in the attention test; however, performance in many individual items did not change. The adherent patients also improved verbal fluency and in the Part B of the Trail making test. The non-adherent group significantly increased the number of mistakes made in the d2 test; other results were non-significant. Conclusion: According to our results, OSA patients' mood, anxiety and certain cognitive domains improved during the one-year therapy with CPAP. Trial Registration Number: NCT03866161.

Early life experiences and adult attachment in obsessive-compulsive disorder. Part 1: Relationships between demographic, clinical, and psychological factors in pharmacoresistant OCD.

OBJECTIVES: Obsessive-compulsive disorder (OCD) has been connected to various psychosocial factors that might influence its onset and course. Developmental factors, such as parenting styles or early adverse experiences, and adult attachment have been listed as examples. However, the research on the interconnections of these factors brought mixed results. The study explores the relationship between demographic, clinical, and selected psychosocial factors and the severity of adult OCD. METHOD: Eighty-seven pharmacoresistant inpatients with OCD were admitted between October 2019 and August 2022 for a 6-week cognitive behavioural therapy inpatient program in the psychotherapeutic department. The participants completed the following scales at the start of the hospitalisation: the self-report Yale-Brown Obsessive-Compulsive Scale (Y-BOCS-SR), Beck Anxiety Inventory (BAI), Beck Depression Inventory (BDI-II), Dissociative Experiences Scale (DES), Childhood Trauma Questionnaire-Short Form (CTQ-SF), PBI (Parental Bonding Instrument), ECR-R (Experiences in Close Relationships - Revised), and a demographic questionnaire. A skilled psychologist administered Mini International Neuropsychiatric Interview (MINI) to confirm the OCD diagnosis and Hamilton Anxiety Rating Scale (HAMA). RESULTS: OCD patients with more severe adverse childhood experiences (ACEs) showed earlier onset of the disorder and more pronounced attachment anxiety, depressive symptoms, and dissociation and subjectively rated the severity of the disorder as more severe. Physical abuse and physical neglect were related to the severity of specific OCD symptoms. Maternal care negatively correlates with clinician-rated anxiety, patient-rated depressive symptoms, and dissociation. The maternal and paternal control positively correlated with patient-rated anxiety and depression. Attachment anxiety negatively correlated with the age of onset and positively with the severity of the clinician-rated anxiety and the patient-rated anxiety, depressive symptoms, and dissociation. CONCLUSIONS: Early adverse experiences, perceived parental styles, and adult attachment anxiety could play a significant role in the symptoms of anxiety, depression, and dissociation. The connection with the specific obsessive-compulsive symptoms is less apparent. Still, adverse childhood events and adult attachment anxiety seem to influence the age of OCD onset.

Early life experiences and adult attachment in obsessive-compulsive disorder. Part 2: Therapeutic effectiveness of combined cognitive behavioural therapy and pharmacotherapy in treatment-resistant inpatients.

OBJECTIVES: Obsessive-compulsive disorder (OCD) is a chronic mental disorder that is often hard to treat with current treatment options. Therapeutic outcomes are predicted by many factors, ranging from biological to psychosocial. Early life experiences and adult attachment influence the effectiveness of the treatment. This study explores their predictive abilities in the combined treatment of adult inpatients with OCD. METHODS: Seventy-seven patients with OCD, diagnosed according to the ICD-10 criteria, were included in the study, out of which 66 patients completed the treatment. All patients were previously unsuccessfully treated with a minimum of two antidepressants for three months each. They were evaluated with rating scales and questionnaires at the start and the end of a six-week hospitalization in a psychotherapeutic department. The treatment approach presented a combination of group cognitive-behavioural therapy and medication. RESULTS: The average severity of OCD, anxiety and depressive symptoms significantly decreased during the inpatient treatment. The improvement in Y-BOCS negatively correlated with the age of onset. The history of emotional abuse and neglect and physical neglect predicted a lower change in anxiety evaluated by a psychologist and perceived maternal care positively correlated with a decrease in anxiety assessed with a rating scale. Adult attachment anxiety predicted a lower decrease in the anxiety measured by the clinician but not the OCD symptomatology. Dissociative symptoms did not significantly predict a change in any of the measures. Comorbid personality disorder did not have a significant impact on therapeutic change. CONCLUSIONS: The early onset of the disorder was the sole predictor of the treatment outcomes regarding specific OCD symptomatology. Selected early adverse experiences, maternal care, and adult attachment anxiety predicted a change in the anxiety symptoms. Future research should focus on mediation and moderation analyses that could help target specific treatment strategies to decrease the impact of these factors.

Colposcopic scoring indexes in the evaluation of cervical lesions with the cytological result of atypical squamous cells, cannot exclude high-grade lesion.

AIM: Colposcopic indexes including Reid index and Swede score were developed to make the colposcopy more objective. The aim of our study was to evaluate the significance of colposcopic indexes in the evaluation of ASC-H cervical lesions. METHODS: We carried out a cross-sectional study in the Clinic of Obstetrics and Gynecology between January 2013 and December 2018. The study included 535 women, from which 66 women had a cytological result ASC-H. Scoring of all colposcopic findings was assessed according to Reid modified index and Swede score and a composite score was determined. Frequency distributions were compared using χ2 /Fisher exact test. Spearman rank correlation coefficient was computed between RCI and Swede score. RESULTS: Sensitivity, specificity, positive and negative predictive value and positive likelihood ratio of modified Reid colposcopic index at a cutoff of ≥4 for the detection of HSIL+ lesions were: 86.11% (95% CI: 70.5-95.3), 83.33% (95% CI: 65.3-94.4), 86.11% (95% CI: 69.7-94.8), 83.33% (95% CI: 64.5-93.7) and 5.17 (95% CI: 2.3-11.6). Swede score with the cutoff value ≥5 showed comparable results to modified Reid index with the increased sensitivity: 94.44% (95% CI: 81.3-99.3). CONCLUSION: ASC-H category represents the trickiest cytological diagnosis as it is underlined with the high risk of severe cervical dysplasia. Evaluating the cervical lesion by the use of colposcopic indices helps the gynecologist to objectively evaluate all the pathologies of uterine cervix. Swede score with the cutoff value 8 also enables a 'see and treat' option in management of atypical squamous cells, cannot exclude high-grade lesions.

Different methylation levels in the KLF4, ATF3 and DLEC1 genes in the myometrium and in corpus uteri mesenchymal tumours as assessed by MS-HRM.

Mesenchymal tumours of the corpus uteri comprise common benign lesions - leiomyomas and very rare malignant variants - sarcomas. It can be difficult to distinguish between the particular types of mesenchymal tumours pre-surgically. Primarily, leiomyomas and the very aggressive leiomyosarcomas can be easily misdiagnosed when using only imaging devices. Therefore, a reliable non-invasive marker for these tumour types would provide greater certitude for patients that the lesion remains benign. Our collection comprises 76 native leiomyomas, an equal number of healthy myometrium samples and 49 FFPE samples of various types of sarcomas. The methylation level was assessed by MS-HRM method and we observed differences in the methylation level between healthy, benign and (semi)malignant tissues in the KLF4 and DLEC1 genes. The mean methylation levels of leiomyomas compared to myometrium and leiomyosarcomas were 70.7% vs. 6.5% vs. 39.6 % (KLF4) and 66.1% vs. 14.08% vs. 37.5% (DLEC1). The ATF3 gene was differentially methylated in leiomyomatous and myometrial tissues with 98.1% compared to 76.6%. The AUC values of the predictive logistic regression model for discrimination between leiomyomas and leiomyosarcomas based on methylation levels were 0.7829 (KLF4) and 0.7719 (DLEC1). Finally, our results suggest that there should be distinct models for the methylation events in benign leiomyomas and sarcomas, and that the KLF4 and DLEC1 genes can be considered potential methylation biomarkers for uterine leiomyomas.

Methylation of CADM1 and MAL together with HPV status in cytological cervical specimens serves an important role in the progression of cervical intraepithelial neoplasia.

Cervical cancer (CC) is the second most common type of cancer affecting the female population. The development of CC takes several years, and involves a precancerous stage known as cervical intraepithelial neoplasia (CIN). A key factor in the development of disease is the human papillomavirus (HPV) infection, which initiates carcinogenesis. Furthermore, CC is also impacted by epigenetic changes such as DNA methylation, which causes activation or exclusion of certain genes, and the hypermethylation of cytosines in promoters, thereby switching off previously active genes. The majority of DNA methylation events occur at cytosine-guanine nucleotides, which in the human genome are known as CpG islands. The aim of the present study was to investigate the methylation levels in intronic sequences of the two tumor suppressor genes cell adhesion molecule 1 (CADM1) and T-lymphocyte maturation associated protein (MAL) using cytological samples and to identify potential biomarkers involved in CIN by pyrosequencing. DNA was isolated from cervical smears from patients with CINs, with healthy patients serving as a control group. Samples were converted by treatment with sodium bisulfite and subsequent pyrosequencing to detect the methylation status of the selected genes. The presence of HPV DNA infection analyzed by the polymerase chain reaction, was detected in each sample. Of the total number of samples (n=91), the present study confirmed the presence of one or two high-risk subtypes of HPV in 39 cases (42.85%) and HPV infection was significantly associated with CIN2+ lesions. For the two genes (MAL and CADM1) the present study confirmed that the median methylation was significantly higher in HPV positive patients [P=0.0097, 95% confidence interval (CI): (-0.030, -0.003)/P=0.0024, 95% CI: (-0.06, -0.01)] when compared with patients negative for HPV DNA infection, and the average methylation was demonstrated to be increased with the degree of cervical lesion. The present study used logistical regression to model the dependence between the case/control statuses (control group vs. Dg. 1-4). The area under the curve values for MAL were: 84% for cervical inflammation, 71% for CIN1, 73.4% for CIN2+ and 77% for squamous cell carcinoma (SCC); and for CADM1 were: 88.6% for cervical inflammation, 68% for CIN1, 80% for CIN2+ and 89% for SCC. The present study confirmed that there were statistically significant differences between the methylation levels of individual CpGs and significantly higher median methylation in patients positive for HPV16/18. CADM1 exhibited higher levels of methylation in almost every study group when compared with MAL during the transition of CIN and appeared to be a promising biomarker for future study.

Different amplification patterns of 3q26 and 5p15 regions in cervical intraepithelial neoplasia and cervical cancer.

PURPOSE: The aim of this study was to evaluate and correlate the amplification of chromosomal regions 3q26 and 5p15 in different cytological and histological subgroups of patients and to compare the sensitivity and specificity of amplification tests with cytology, colposcopy and HPV status. METHODS: The work was conducted at the Department of Obstetrics and Gynaecology in cooperation with the Institute of Pathological Anatomy, JFM CU in Martin and UNM during years 2013-2016. Prospective longitudinal study included 131 patients. We focused on the FISH diagnosis of the amplification of regions encoding the components of telomerase enzyme (3q26, 5p15) in cytology specimens. We manually evaluated 100 atypical cells per slide and analysed the amplification patterns. Correlations between cytological, histological, HPV DNA results and amplification patterns of chromosomal regions 3q26 and 5p15 were analysed by chi-squared test and non-parametric Man - Whitney U test. RESULTS: The results showed that the amplification of chromosomal regions increases with the degree of dysplasia toward the invasive disease (p < 0.001). Whereas the increase in the amplification of 3q26 is noticeable already at CIN 2 + lesions (p < 0.01), 5p15 amplification is shifted up toward CIN 3/CIS (p < 0.001) and cervical cancer. Amplification of selected regions correlated with each other and also with hrHPV-positive status (p < 0.01). CONCLUSION: The analysis of the amplification of 3q26 and 5p15 regions may serve in the future for the differential diagnosis of cervical lesions and to determine their malignant potential. High specificity of these tests can improve the excellent sensitivity of HPV DNA test.

Molecular and clinical attributes of uterine leiomyomas.

Uterine leiomyomas, also called uterine fibroids or myomas, represent one of the most common benign tumour types in women of a fertile age. Leiomyomas arise due to transformation of the layer of smooth muscle cells of corpus uteri - the myometrium. Despite frequent occurrence of this disease, the molecular mechanisms behind the origin and development of leiomyomas are still relatively unknown. Most predisposed are obese women and women of African origin. In more than half of cases, leiomyomas remain asymptomatic. Genetic factors also have an important impact on the development of these hormone-dependent tumours. However, the clinical and molecular characteristics of familiar and sporadic leiomyomas can widely differ. The main reason is the heterogeneity of this disease and the abundance of factors which can underlie their tumourigenesis. Clinical diagnosis of uterine leiomyomas without surgical interference can be hindered in the case of small, mostly submucosal leiomyomas or if it is necessary to avoid potential malignancy of tumour. Also, medical treatment of uterine leiomyomas cannot be nowadays considered sufficient with many medical agents still being tested only within clinical research. The main goal of this article is to summarise known facts about the aetiology of leiomyomas, risk factors that contribute to their development, known molecular-genetic aberrations connected with the presence of leiomyomas as well as the possibilities of their diagnosis and treatment.

DNA methylation as mechanism of apoptotic resistance development in endometrial cancer patients.

DNA methylation is a significant epigenetic modification which plays a key role in regulation of gene expression and influences functional changes in endometrial tissue. Aberrant DNA methylation changes result in deregulation of important apoptotic proteins during endometrial carcinogenesis and apoptosis resistance development. Evading apoptosis is still a major problem in the successful treatment of endometrial cancer patients. The aim of our study was to examine the promoter DNA methylation changes in 22 apoptosis-associated genes in endometrioid endometrial cancer patients, precancerous lesions and healthy tissue from various normal menstrual cycle phases using a unique pre-designed methylation platform. We observed as the first a significant difference in promoter DNA methylation status in genes: BCL2L11 (p < 0.001), CIDEB (p < 0.03) and GADD45A (p < 0.05) during endometrial carcinogenesis and BIK gene (p < 0.03) in different phases of normal menstrual cycle. The results of our study indicate that deregulation of mitochondrial apoptotic pathway can considerably contributes to the apoptosis resistance development and may be helpful in identifying of new potent biomarkers in endometrial cancer.

The examination of superior mesenteric artery circulation in fetus during pregnancy.

OBJECTIVE: The aim of the study was a comprehensive examination of the circulation of superior mesenteric artery in different weeks of pregnancy using Doppler ultrasound examination. METHODS: The study was conducted at the Clinic of Obstetrics and Gynecology, Jessenius Faculty of Medicine, Comenius University, Slovakia between the years 2008 and 2012. Dynamics and changes in superior mesenteric artery circulation were studied in 856 women. Results of Doppler examination were processed and subsequently evaluated for each gestational week separately. RESULTS: We found that changes in resistance index of superior mesenteric artery are slower and have a tendency to decrease as a consequence of a gradual decrease in vascular resistance. The changes in pulsatility index are different in nature; there is a tendency for there are to be repeating periods of higher and then lower values, which are subject to a more pronounced dependence on flow rate, especially end diastolic velocity, and Vmean. CONCLUSIONS: This article and its results bring a new, comprehensive view not only of physiological changes in the splanchnic circulation, but also of changes that may be subject to a certain extent, not only to the sex, but primarily to the weight of the fetus, which, of course, indirectly reflects its metabolic and respiratory demands.

Survival and risk factors associated with uterine sarcomas and carcinosarcomas in stage I and II.

OBJECTIVE: Uterine sarcomas are rare mesodermal malignant tumors with an incidence between 0.5 and 3.3 cases per 100,000 females per year. Most sarcomas are aggressive tumors leading to poor overall survival rates and only limited therapeutic options. The aim of this study was to evaluate the risk factors for uterine sarcomas and carcinosarcomas, and to identify the factors influencing the survival rate. SUBJECTS AND METHODS: We conducted a retrospective study with twenty-nine patients who were diagnosed with uterine sarcoma and thirty-four patients with carcinosarcoma between the years 1990 and 2006 at the Oncogynecologic center at the University Hospital in Martin, Slovakia. We focused on the analysis of the risk factors and survival rate of early stages I and II. RESULTS: We confirmed highly statistically significant values for the inverse correlation between survival and tumor size, positive lymph nodes, high mitotic activity, vascular invasion, positive peritoneal cytology, elevated CA-125, smoking and BMI in sarcoma and carcinosarcoma group (p<0.001 for all factors). The use of lymphadenectomy had no effect on survival of all patients. DISCUSSION: Sarcomas and carcinosarcomas are aggressive tumors leading to poor overall survival rates and only limited therapeutic options. As there is no consensus on specific treatment, an individual approach based on evaluation of known risk factors is essential.

The analysis of linear and monomethylalkanes in exhaled breath samples by GC×GC-FID and GC-MS/MS.

A new arrangement of the INCAT (inside needle capillary adsorption trap) device with Carbopack X and Carboxen 1000 as sorbent materials was applied for sampling, preconcentration and injection of C6C19n-alkanes and their monomethyl analogs in exhaled breath samples. For the analysis both GC-MS/MS and GC×GC-FID techniques were used. Identification of the analytes was based on standards, measured retention indices and selective SRM transitions of the individual isomers. The GC-MS/MS detection limits were in the range from 2.1 pg for n-tetradecane to 86 pg for 5-methyloctadecane. The GC×GC-FID detection limits ranged from 19 pg for n-dodecane to 110 pg for 3-methyloctane.

Amplification of TERT and TERC genes in cervical intraepithelial neoplasia and cervical cancer.

OBJECTIVES: Telomerase is activated in various stages of oncogenesis. For cervical cancer, telomerase is already active in precancerous lesions. In our study we focused on the analysis of the amplification patterns of telomerase genes TERT and TERC. DESIGN AND SETTING: We included 39 patients in our study between January 2012 and April 2013. Each patient underwent a classical gynaecological examination and a colposcopy. During the colposcopic examination we collected material for a Pap smear, HPV DNA test (HC2) and LBC (LiquiPrep™), and performed punch biopsies for histopathological evaluation. Residual cytologic sample was hybridized with the FISH probe and telomerase genes were analysed. RESULTS: The amplification of the TERT gene showed us a very similar amplification pattern as TERC and gradually corresponded with both histolopathological (p<0.001) and cytopathological findings (p<0.001). The specificity and sensitivity of TERC gene amplification for the detection of CIN2+ lesions (cut off value 2.3) was 88.2% and 95.5% respectively (PPV 91.3%, NPV 93.8%). CONCLUSIONS: We identified increasing amplification pattern of telomerase genes in cervical lesions. According to our results telomerase genes could help in the future to determine the malignant potential of cervical lesions and could be tested together with cytology and HPV DNA in order to obtain the highest combined sensitivity and specificity for CIN2+ lesion detection.

Amplification of 3q26 and 5p15 regions in cervical intraepithelial neoplasia.

OBJECTIVE: To analyze different amplification patterns of 3q26 and 5p15 regions in low-grade and high-grade cervical intraepithelial neoplasia. DESIGN: Experimental research. SETTING: Department of Obstetrics and Gynecology at a medical faculty in Slovakia. POPULATION: A group of 83 patients referred for colposcopic examination. METHODS: Amplification of 3q26 and 5p15 regions was analyzed on the 100 most atypical cells from a cervical cytology slide by fluorescent in situ hybridization using a multicolor hybridization probe. Chi-squared and Man-Whitney U-tests were used for statistical analysis. MAIN OUTCOME MEASURES: Liquid-based cytology samples and biopsy samples obtained during colposcopic examination correlated with high-risk human papillomavirus status and with amplification patterns of selected regions analyzed by fluorescent in situ hybridization. RESULTS: The number of cells with 3q26 and 5p15 gain rises with the severity of the lesion p < 0.01. The sensitivity of 3q26 amplification for CIN2+ lesions was 72.1% (95% confidence interval 56.3-84.7) and specificity was 90.0% (95% confidence interval 76.3-97.1). The sensitivity of 5p15 amplification for CIN2+ lesions was 69.8% (95% confidence interval 53.9-82.8) and specificity was 85.0% (95% confidence interval 70.2-94.3). CONCLUSION: Evaluation of telomerase components can help in differential diagnosis of low-grade and high-grade cervical lesions and in individualized management of these patients.

Direct silylation of Trypanosoma brucei metabolites in aqueous samples and their GC-MS/MS analysis.

A simple two-step method for the derivatization of polar compounds (lactate, alanine, glycerol, succinate and glucose) using hexamethyldisilazane (HMDS) and N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) was developed. This method allows direct derivatization of aqueous samples wihout sample pretreatment. The method was used for the analysis of the metabolites of the unicellular organism Trypanosoma brucei. The limits of detection by GC-MS/MS analysis were in the range of 0.02 mg L(-1) for glucose to 0.85 mg L(-1) for lactate.

Detection of methylation of the promoter region of the MAL and CADM1 genes by pyrosequencing in cervical carcinoma.

OBJECTIVE: Cervical cancer is the second most common cancer disease affecting the female population. A key factor in development of the disease is the human papillomavirus infection (HPV). The disease is also impacted by epigenetic changes such as DNA methylation, which causes activation or exclusion of certain genes, and simultaneously the hypermethylation of cytosines in the promoters and turn-off of previously active genes occur. In this study, we focused on the introduction of pyrosequencing for the detection of DNA methylation of the selected CADM1 and MAL genes. METHODS: DNA was isolated from cytological cervical smear of patients with different types of dysplasia [L-SIL (n=14), ASC-US (n=15), H-SIL (n=1)] and four control samples from healthy women. Prepared samples were further analyzed by bisulfite conversion and subsequent pyrosequencing (Pyromark Q96 ID, Qiagen, Germany). We examined the extent of methylation of CpG islands and as control samples of this method we used a fully methylated and unmethylated DNA. Methylation level (Met level) from each sample was quantified as the mean value [sum of all methylated CpG islands in %/total number of CpG islands (MAL n=4; CADM1 n=3)]. RESULTS: In total, 30 clinical samples and 4 control samples from healthy women were analyzed. By means of the analysis of the CADM1promoter region, the values of the Met level were obtained [fully methylated DNA (94.83 and 88); completely unmethylated DNA (0 and 0); and control samples from healthy patients (6.825 and 0.825), L-SIL (2.107 and 2.778), ASC-US (7.313 and 3.626), H-SIL (0 and 0)]. By means of the analysis of the MAL promoter region, the values of Met level were obtained [fully methylated DNA (53.25); completely unmethylated DNA (0.875); and control samples from healthy patients (2.925), L-SIL (1.517), ASC-US (2.833), and H-SIL (4)]. CONCLUSION: We introduced a pyrosequencing method for quantification of methylation of CADM1, MAL promoter regions, and detected methylations in clinical samples and also some basal methylation in healthy women.

Promoter hypermethylation of the tumor-suppressor genes RASSF1A, GSTP1 and CDH1 in endometrial cancer.

Endometrial cancer is a common gynecological malignancy with a good prognosis in early stages of the disease. The CpG island in the promoter region of tumor-suppressor genes are frequently methylated in various types of human cancers. In the present study, we examined the methylation status of the GSTP1, CDH1 and RASSF1A genes in endometrioid endometrial cancer (EEC), endometrial complex hyperplasia (EHP) and healthy endometrium with the aim to identify correlations between promoter hypermethylation, disease risk and clinicopathological parameters. A nested two-stage methylation-specific PCR (MSP) was performed to analyze the promoter CpG methylation status of GSTP1, CDH1 and RASSF1A genes in the population studied. A total of 92 subjects were initially included in the study of which 41 EEC, 19 EHP and 20 controls were processed for final analyses. A significant difference was found between the study groups and the presence of promoter CpG hypermethylation status in the GSTP1 (p<0.05) and RASSF1A (p<0.0001) genes. RASSF1A, GSTP1 and CDH1 gene promoter methylation was present in 85.4, 68.3 and 31.4% of EEC samples when compared to that in the controls with 30.0, 35.0 and 20.0%, respectively. CpG methylation of all three investigated tumor-suppressor genes was found in 12.2% of EEC patients, in 4.2% of EHP patients and in 3.7% of the controls, respectively. Positive findings for the promoter methylation of two investigated genes were found in 48.7% of EEC patients, 26.0% of EHP patients and in 18.5% of the controls. With regard to histopathological variables and CpG methylation, we found significant correlations between the RASSF1A and GSTP1 genes and higher tumor grade, deeper myometrial invasion and positive metastatic involvement of pelvic lymph nodes. No associations were noted between promoter hypermethylation of the CDH1 gene and biological features of the endometrial cancer cases. The results indicate that aberrant CpG methylation of the promoter region in the GSTP1 and RASSF1A tumor-suppressor genes is an important event in carcinogenesis of endometrial cancer and may have an impact on tumor aggressiveness. Finally, the present study suggests that epigenetic alterations may be of diagnostic value for the better clinical management of premalignant endometrial lesions.

Fungal neuroinfections and fungaemia: unexpected increase of mortality from invasive fungal infections in 2005-2011 in comparison to 1989-1998: analysis of 210 cases.

OBJECTIVE: In this short communication we compared the data of fungaemia cases in Slovak hospitals from 1989-1998 published in 1999-2000 with data from 2005-2011. METHODS: Risk factors, etiology and outcome of fungaemia between two periods (1989-1998 vs. 2005-2011) were compared and risk factors for death assessed by univariate analysis (CDC 2006 Statistical Package). RESULTS: In comparison to 1989-1998 when only amphotericin B and fluconazole has been used (55%), in 2005-2011 only 35.2% patients received FLU, but 26.4% received voriconazole, 22% caspofungin and anidulafungin and about 6.6% lipid formulations of Amphotericin B. In etiology, while in 1989-1998 only 37.1% (115/310) represented non-albicans Candida (NAC) and non-Candida yeasts in 2005-2011 already reached 63.7%. The significant increase of breakthrough fungaemia may be a sign of inappropriate empiric therapy.