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An evaluation of SOX2 and hTERC gene amplifications as screening markers in oral and oropharyngeal squamous cell carcinomas
Nadja Kokalj-Vokač, Bogdan Čizmarevič, Andreja Zagorac, Boris Zagradišnik, Boštjan Lanišnik, izvirni znanstveni članek

Opis: Background: Oral and oropharyngeal squamous cell carcinomas (OSCC) are among the most common cancers. The poor survival rate among oral cancer patients can be attributed to several factors, one of them being lack of early detection. A key approach to this problem would be to detect potentially malignant lesion at their early stage. Using the FISH technique, oral brush cytology slides can be an easy and rapid screening approach for malignant cell detection. The present study was designed to detect hTERC and SOX2 amplifications in OSSC exfoliative tumor cells and evaluate whether those two gene amplifications might serve as a supportive biomarker in early detection and diagnosis of oral and oropharyngeal SCC. Results: Brush biopsies were collected from exophytic and exulcerated oral and oropharyngeal lesions of the oral cavity of 71 patients and 22 healthy controls. FISH techniques using a TERC-specific DNA probe and a SOX2 DNA specific probe both combined with a centromere 3-specific control probe was performed on the cytology slides. A 100 squamous epithelial cell nuclei of the smears per slide were analysed. As abnormal FISH pattern were considered amplified and polyploid patterns. From 71 brush biopsies of oropharynx and other locations in oral cavity analysed by FISH 49 were considered to be abnormal (69%). The over representation of polyploidy and/or TERC/SOX2 amplification in tumour samples was statistically significant when compared to controls (p = 0.01). Conclusion: SOX2 and TERC gene amplifications are common in all squamous cell carcinomas and their detection in early stages could be crucial for early detection and more accurate prognosis. Our study strongly suggests that early detection by FISH on cytobrushed samples could be a possible non-invasive screening method even before a tissue biopsy is performed.
Ključne besede: SOX and hTERC gene amplifications, brush biopsy, Oral, oropharyngeal squamous cell carcinoma
Objavljeno: 29.06.2017; Ogledov: 68; Prenosov: 0
.pdf Polno besedilo (1,28 MB)

Comorbidities and characteristics of coronary heart disease patients: their impact on health-related quality of life
Ksenija Tušek-Bunc, Davorina Petek, 2016, izvirni znanstveni članek

Opis: Background: Patients with coronary heart disease (CHD) commonly present with more than one comorbid condition, contributing to poorer health-related quality of life (HRQoL). The aim of our study was to identify the associations between HRQoL and patient characteristics, vascular comorbidities and anxiety/depression disorders. Methods: This observational study was conducted in 36 family medicine practices selected by random stratified sampling from all regions of Slovenia. HRQoL was assessed using the European Quality of Life - 5 Dimensions (EQ-5D) questionnaire and EQ Visual Analogue Scale (EQ-VAS). The associations between HRQoL and patient characteristics stratified by demographics, vascular comorbidities, health services used, their assessment of chronic illness care, and anxiety/depression disorders were identified by ordinal logistic regression and linear regression models. Results: The final sample included 423 CHD patients with a mean age of 68.0 ± SD 10.8 years; 35.2% were female. Mean EQ-VAS score was 58.6 ± SD 19.9 (median: 60 with interquartile range of 45–75), and mean EQ-5D index was 0.60 ± SD 0.19 (median: 0.56 with interquartile range of 0.41–0.76). The statistically significant predictors of a lower EQ-VAS score were higher family physician visit frequency, heart failure (HF) and anxiety/depression disorders (R² 0.240; F = 17.368; p < 0.001). The statistically significant predictor of better HRQoL, according to EQ-5D was higher patient education, whereas higher family physician visit frequency, HF and peripheral artery disease (PAD) were predictors of poorer HRQoL (Nagelkerke R 2 = 0.298; χ 2 = 148.151; p < 0.001). Conclusions: Results of our study reveal that comorbid conditions (HF and PAD), family physician visit frequency and years in education are significant predictors of HRQoL in Slovenian CHD patients.
Ključne besede: coronary heart disease patient, health-related quality of life, vascular comorbidities, anxiety/depression disorders
Objavljeno: 29.06.2017; Ogledov: 64; Prenosov: 0
.pdf Polno besedilo (407,23 KB)

Vasopressin improves outcome in out-of-hospital cardiopulmonary resuscitation of ventricular fibrillation and pulseless ventricular tachycardia: a observational cohort study
Štefek Grmec, Štefan Mally, 2006, izvirni znanstveni članek

Opis: Introduction: An increasing body of evidence from laboratory and clinical studies suggests that vasopressin may represent a promising alternative vasopressor for use during cardiac arrest and resuscitation. Current guidelines for cardiopulmonary resuscitation recommend the use of adrenaline (epinephrine), with vasopressin considered only as a secondary option because of limited clinical data. Method: The present study was conducted in a prehospital setting and included patients with ventricular fibrillation or pulseless ventricular tachycardia undergoing one of three treatments: group I patients received only adrenaline 1 mg every 3 minutes; group II patients received one intravenous dose of arginine vasopressine (40 IU) after three doses of 1 mg epinephrine; and patients in group III received vasopressin 40 IU as first-line therapy. The cause of cardiac arrest (myocardial infarction or other cause) was established for each patient in hospital. Results: A total of 109 patients who suffered nontraumatic cardiac arrest were included in the study. The rates of restoration of spontaneous circulation and subsequent hospital admission were higher in vasopressin-treated groups (23/53 [45%] in group I, 19/31 [61%] in group II and 17/27 [63%] in group III). There were also higher 24-hour survival rates among vasopressin-treated patients (P < 0.05), and more vasopressin-treated patients were discharged from hospital (10/51 [20%] in group I, 8/31 [26%] in group II and 7/27 [26%] group III; P = 0.21). Especially in the subgroup of patients with myocardial infarction as the underlying cause of cardiac arrest, the hospital discharge rate was significantly higher in vasopressin-treated patients (P < 0.05). Among patients who were discharged from hospital, we found no significant differences in neurological status between groups. Conclusion: The greater 24-hour survival rate in vasopressin-treated patients suggests that consideration of combined vasopressin and adrenaline is warranted for the treatment of refractory ventricular fibrillation or pulseless ventricular tachycardia. This is especially the case for those patients with myocardial infarction, for whom vasopressin treatment is also associated with a higher hospital discharge rate.
Ključne besede: vasopressin, antidiuretic hormone, ADH, out-of-hospital cardiopulmonary resuscitation, ventricular fibrillation, pulseless ventricular tachycardia
Objavljeno: 29.06.2017; Ogledov: 39; Prenosov: 0
.pdf Polno besedilo (159,29 KB)

The effect of carbon dioxide on near-death experiences in out-of-hospital cardiac arrest survivors: a prospective observational study
Zalika Klemenc-Ketiš, Janko Kersnik, Štefek Grmec, 2010, izvirni znanstveni članek

Opis: Introduction: Near-death experiences (NDEs) are reported by 11-23% of cardiac arrest survivors. Several theories concerning the mechanisms of NDEs exist - including physical, psychological, and transcendental reasons - but so far none of these has satisfactorily explained this phenomenon. In this study, we investigated the effect of partial pressures of O2 and CO2, and serum levels of Na and K on the occurrence of NDEs in out-of-hospital cardiac arrest survivors. Methods: A prospective observational study was conducted in the three largest hospitals in Slovenia. Fifty-two consecutive patients (median age 53.1 years, 42 males) after out-of-hospital cardiac arrest were included. The presence of NDEs was assessed with a self-administered Greyson's NDE scale. The initial partial pressure of end-tidal CO2, the arterial blood partial pressures of O2 and CO2 and the levels of Na and K in venous blood were analysed and studied. Univariate analyses and multiple regression models were used. Results: NDEs were reported by 11 (21.2%) of the patients. Patients with higher initial partial pressures of end-tidal CO2 had significantly more NDEs (P < 0.01). Patients with higher arterial blood partial pressures of CO2 had significantly more NDEs (P = 0.041). Scores on a NDE scale were positively correlated with partial pressures of CO2 (P = 0.017) and with serum levels of potassium (P = 0.026). The logistic regression model for the presence of NDEs (P = 0.002) explained 46% of the variance and revealed higher partial pressures of CO2 to be an independent predictor of NDEs. The linear regression model for a higher score on the NDE scale (P = 0.001) explained 34% of the variance and revealed higher partial pressures of CO2, higher serum levels of K, and previous NDEs as independent predictors of the NDE score. Conclusions: Higher concentrations of CO2 proved significant, and higher serum levels of K might be important in the provoking of NDEs. Since these associations have not been reported before, our study adds novel information to the field of NDEs phenomena.
Ključne besede: near-death experience, cardiac arrest, survivors
Objavljeno: 29.06.2017; Ogledov: 36; Prenosov: 0
.pdf Polno besedilo (301,05 KB)

The dynamic pattern of end-tidal carbon dioxide during cardiopulmonary resuscitation - difference between asphyxial cardiac arrest and ventricular fibrillation/pulseless ventricular tachycardia cardiac arrest
Katja Lah, Miljenko Križmarić, Štefek Grmec, 2011, izvirni znanstveni članek

Opis: Introduction: Partial pressure of end-tidal carbon dioxide (PetCO2) during cardiopulmonary resuscitation (CPR) correlates with cardiac output and consequently has a prognostic value in CPR. In our previous study we confirmed that initial PetCO2 value was significantly higher in asphyxial arrest than in ventricular fibrillation/pulseless ventricular tachycardia (VF/VT) cardiac arrest. In this study we sought to evaluate the pattern of PetCO2 changes in cardiac arrest caused by VF/VT and asphyxial cardiac arrest in patients who were resuscitated according to new 2005 Guidelines. Methods: The study included two cohorts of patients: cardiac arrest due to asphyxia with initial rhythm asystole or pulseless electrical activity (PEA), and cardiac arrest due to arrhythmia with initial rhythm VF or pulseless VT. PetCO2 was measured for both groups immediately after intubation and repeatedly every minute, both for patients with or without return of spontaneous circulation (ROSC). We compared the dynamic pattern of PetCO2 between groups. Results: Between June 2006 and June 2009 resuscitation was attempted in 325 patients and in this study we included 51 patients with asphyxial cardiac arrest and 63 patients with VF/VT cardiac arrest. The initial values of PetCO2 were significantly higher in the group with asphyxial cardiac arrest (6.74 +/- 4.22 kPa versus 4.51 +/- 2.47 kPa; P = 0.004). In the group with asphyxial cardiac arrest, the initial values of PetCO2 did not show a significant difference when we compared patients with and without ROSC (6.96 +/- 3.63 kPa versus 5.77 +/- 4.64 kPa; P = 0.313). We confirmed significantly higher initial PetCO2 values for those with ROSC in the group with primary cardiac arrest (4.62 +/- 2.46 kPa versus 3.29 +/- 1.76 kPa; P = 0.041). A significant difference in PetCO2 values for those with and without ROSC was achieved after five minutes of CPR in both groups. In all patients with ROSC the initial PetCO2 was again higher than 1.33 kPa. Conclusions: The dynamic pattern of PetCO2 values during out-of-hospital CPR showed higher values of PetCO2 in the first two minutes of CPR in asphyxia, and a prognostic value of initial PetCO2 only in primary VF/VT cardiac arrest. A prognostic value of PetCO2 for ROSC was achieved after the fifth minute of CPR in both groups and remained present until final values. This difference seems to be a useful criterion in prehospital diagnostic procedures and attendance of cardiac arrest.
Ključne besede: end-tidal carbon dioxide, ETCO2, cardiopulmonary resuscitation, CPR, asphyxial cardiac arrest, ventricular fibrillation, pulseless ventricular tachycardia, cardiac arrest
Objavljeno: 29.06.2017; Ogledov: 34; Prenosov: 1
.pdf Polno besedilo (1,58 MB)

Combination of lung ultrasound (a comet-tail sign) and N-terminal pro-brain natriuretic peptide in differentiating acute heart failure from chronic obstructive pulmonary disease and asthma as cause of acute dyspnea in prehospital emergency setting
Gregor Prosen, Petra Klemen, Matej Strnad, Štefek Grmec, 2011, izvirni znanstveni članek

Opis: Introduction: We studied the diagnostic accuracy of bedside lung ultrasound (the presence of a comet tail sign), N-terminal pro-brain natriuretic peptide (NT-proBNP), and clinical assessment (modified Boston criteria) in differentiating heart failure (HF)- related acute dyspnea from pulmonary (COPD/asthma) related acute dyspnea in the prehospital setting. Methods: Prospective study was performed at the Center for Emergency Medicine Maribor, Slovenia, between July 2007 and April 2010. Two groups of patients were compared: HF-related acute dyspnea group (n = 129) vs pulmonary-related (asthma/COPD) acute dyspnea group (n = 89). All patients underwent lung ultrasound examination, along with basic laboratory, rapid NT-proBNP testing and chest X-ray. Results: Ultrasound comet tail sign has 100% sensitivity, 95% specificity, 100% negative predictive value (NPV) and 96% positive predictive value (PPV) for the diagnosis of HF. NT-proBNP (cut-off point 1000 pg/ml) has 92% sensitivity, 89% specificity, 86% NPV and 90% PPV. Boston modified criteria have 85% sensitivity, 86% specificity, 80% NPV and 90% PPV. Comparing the three methods, we found significant differences between ultrasound sign vs NT-proBNP (P<0.05) and Boston modified criteria (P<0.05). Combination of ultrasound sign and NT-proBNP has 100% sensitivity, 100% specificity, 100% NPV and 100% PPV. With ultrasound we can exclude HF in patients with pulmonary related dyspnea who have positive NT-proBNP (> 1000 pg/ml) and previous history of HF. Conclusions: Ultrasound comet tail sign alone or in combination with NT-proBNP has a high diagnostic accuracy in differentiating between acute HF and COPD/asthma causes of acute dyspnea in prehospital emergency setting.
Ključne besede: lungs, ultrasound, N-terminal pro-brain natriuretic peptide, acute heart failure, chronic obstructive pulmonary disease, asthma, acute dyspnea
Objavljeno: 29.06.2017; Ogledov: 50; Prenosov: 0
.pdf Polno besedilo (420,76 KB)

TMPRSS2:ERG gene aberrations may provide insight into pT stage in prostate cancer
Zoran Krstanoski, Nadja Kokalj-Vokač, Andreja Zagorac, Boris Pospihalj, Miha Munda, Sašo Džeroski, Rastko Golouh, 2016, izvirni znanstveni članek

Opis: Background: TMPRSS2:ERG gene aberration may be a novel marker that improves risk stratification of prostate cancer before definitive cancer therapy, but studies have been inconclusive. Methods: The study cohort consisted of 202 operable prostate cancer Slovenian patients who underwent laparoscopic radical prostatectomy. We retrospectively constructed tissue microarrays of their prostatic specimens for fluorescence in situ hybridization, with appropriate signals obtained in 148 patients for subsequent statistical analyses. Results: The following genetic aberrations were found: TMPRSS2:ERG fusion, TMPRSS2 split (a non-ERG translocation) and ERG split (an ERG translocation without involvement of TMPRSS2). TMPRSS2:ERG gene fusion happened in 63 patients (42 %), TMPRSS2 split in 12 patients and ERG split in 8 patients. Association was tested between TMPRSS2:ERG gene fusion and several clinicopathological variables, i.e., pT stage, extended lymph node dissection status, and Gleason score, correcting for multiple comparisons. Only the association with pT stage was significant at p = 0.05: Of 62 patients with pT3 stage, 34 (55 %) had TMPRSS2:ERG gene fusion. In pT3 stage patients, stronger (but not significant) association between eLND status and TMPRSS2:ERG gene fusion was detected. We detected TMPRSS2:ERG gene fusion in 64 % of the pT3 stage patients where we did not perform an extended lymph node dissection. Conclusions: Our results indicate that it is possible to predict pT3 stage at final histology from TMPRSS2:ERG gene fusion at initial core needle biopsy. FISH determination of TMPRSS2:ERG gene fusion may be particularly useful for patients scheduled to undergo a radical prostatectomy in order to improve oncological and functional results.
Ključne besede: FISH, predicting pT stage, radical prostatectomy, prostate cancer, TMPRSS2:ERG fusion
Objavljeno: 29.06.2017; Ogledov: 64; Prenosov: 0
.pdf Polno besedilo (1,06 MB)

Typing Clostridium difficile strains based on tandem repeat sequences
N Henning Zaiß, Maja Rupnik, Ed J Kuijper, Celine Harmanus, Dolf Michielsen, Koen Janssens, Ulrich Nübel, 2009, izvirni znanstveni članek

Opis: Background: Genotyping of epidemic Clostridium difficile strains is necessary to track their emergence and spread. Portability of genotyping data is desirable to facilitate inter-laboratory comparisons and epidemiological studies. Results: This report presents results from a systematic screen for variation in repetitive DNA in the genome of C. difficile. We describe two tandem repeat loci, designated 'TR6' and 'TR10', which display extensive sequence variation that may be useful for sequence-based strain typing. Based on an investigation of 154 C. difficile isolates comprising 75 ribotypes, tandem repeat sequencing demonstrated excellent concordance with widely used PCR ribotyping and equal discriminatory power. Moreover, tandem repeat sequences enabled the reconstruction of the isolates' largely clonal population structure and evolutionary history. Conclusion: We conclude that sequence analysis of the two repetitive loci introduced here may be highly useful for routine typing of C. difficile. Tandem repeat sequence typing resolves phylogenetic diversity to a level equivalent to PCR ribotypes. DNA sequences may be stored in databases accessible over the internet, obviating the need for the exchange of reference strains.
Ključne besede: Clostridium difficile, microbiology, strain typing
Objavljeno: 29.06.2017; Ogledov: 33; Prenosov: 1
.pdf Polno besedilo (792,24 KB)

The LexA regulated genes of the Clostridium difficile
Beata Maria Walter, Maja Rupnik, Vesna Hodnik, Gregor Anderluh, Bruno Dupuy, Nejc Paulič, Darja Žgur-Bertok, Matej Butala, 2014, izvirni znanstveni članek

Opis: Background: The SOS response including two main proteins LexA and RecA, maintains the integrity of bacterial genomes after DNA damage due to metabolic or environmental assaults. Additionally, derepression of LexA-regulated genes can result in mutations, genetic exchange and expression of virulence factors. Here we describe the first comprehensive description of the in silico LexA regulon in Clostridium difficile, an important human pathogen. Results: We grouped thirty C. difficile strains from different ribotypes and toxinotypes into three clusters according to lexA gene/protein variability. We applied in silico analysis coupled to surface plasmon resonance spectroscopy (SPR) and determined 16 LexA binding sites in C. difficile. Our data indicate that strains within the cluster, as defined by LexA variability, harbour several specific LexA regulon genes. In addition to core SOS genes: lexA, recA, ruvCA and uvrBA, we identified a LexA binding site on the pathogenicity locus (PaLoc) and in the putative promoter region of several genes involved in housekeeping, sporulation and antibiotic resistance. Conclusions: Results presented here suggest that in C. difficile LexA is not merely a regulator of the DNA damage response genes but also controls the expression of dozen genes involved in various other biological functions. Our in vitro results indicate that in C. difficile inactivation of LexA repressor depends on repressor`s dissociation from the operators. We report that the repressor`s dissociation rates from operators differentiate, thus the determined LexA-DNA dissociation constants imply on the timing of SOS gene expression in C. difficile.
Ključne besede: Clostridium difficile, antibiotic resistance, toxin regulation, SOS system, surface plasmon resonance, SPR, LexA repressor
Objavljeno: 29.06.2017; Ogledov: 35; Prenosov: 1
.pdf Polno besedilo (1,16 MB)

International Clostridium difficile animal strain collection and large diversity of animal associated strains
Sandra Janežič, Valerija Tkalec, Bart Pardon, Alexander Indra, Branko Kokotovic, Jose Luis Blanco, Christian Seyboldt, Rodriguez Diaz, Ian Poxton, Vincent Perreten, Ilenia Drigo, Alena Jiraskova, Matjaž Ocepek, Scott Weese, Glenn Songer, Mark Wilcox, Maja Rupnik, 2014, izvirni znanstveni članek

Opis: Background: Clostridium difficile is an important cause of intestinal infections in some animal species and animals might be a reservoir for community associated human infections. Here we describe a collection of animal associated C. difficile strains from 12 countries based on inclusion criteria of one strain (PCR ribotype) per animal species per laboratory. Results: Altogether 112 isolates were collected and distributed into 38 PCR ribotypes with agarose based approach and 50 PCR ribotypes with sequencer based approach. Four PCR ribotypes were most prevalent in terms of number of isolates as well as in terms of number of different host species: 078 (14.3% of isolates; 4 hosts), 014/020 (11.6%; 8 hosts); 002 (5.4%; 4 hosts) and 012 (5.4%; 5 hosts). Two animal hosts were best represented; cattle with 31 isolates (20 PCR ribotypes; 7 countries) and pigs with 31 isolates (16 PCR ribotypes; 10 countries). Conclusions: This results show that although PCR ribotype 078 is often reported as the major animal C. difficile type, especially in pigs, the variability of strains in pigs and other animal hosts is substantial. Most common human PCR ribotypes (014/020 and 002) are also among most prevalent animal associated C. difficile strains worldwide. The widespread dissemination of toxigenic C. difficile and the considerable overlap in strain distribution between species furthers concerns about interspecies, including zoonotic, transmission of this critically important pathogen.
Ključne besede: Clostridium difficile, animals, ribotyping, geographic distribution, strain collection
Objavljeno: 29.06.2017; Ogledov: 35; Prenosov: 1
.pdf Polno besedilo (985,16 KB)

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