Association of HOTAIR, MIR155HG, TERC, miR-155, -196a2, and -146a Genes Polymorphisms with Papillary Thyroid Cancer Susceptibility and Prognosis.

Polymorphisms in long non-coding RNA and microRNA genes may play a significant role in the susceptibility and progression of papillary thyroid carcinoma (PTC). The current study investigates the polymorphisms HOTAIR rs920778, MIR155HG rs1893650, TERC rs10936599, miR-155 rs767649, miR-196a2 rs11614913 and miR-146a rs2910164 in 102 PTC patients and 106 age- and sex-matched controls of the Caucasian Serbian population, using real-time PCR. We observed differences in genotype distributions of the HOTAIR rs920778 (p = 0.016) and MIR155HG rs1893650 (p = 0.0002) polymorphisms between PTC patients and controls. HOTAIR rs920778 was associated with increased PTC susceptibility (adjusted OR = 1.497, p = 0.021), with the TT variant genotype increasing the risk compared to the CC genotype (OR = 2.466, p = 0.012) and C allele carriers (CC + CT) (OR = 1.585, p = 0.006). The HOTAIR rs920778 TT genotype was associated with lymph node metastasis (p = 0.022), tumor recurrence (p = 0.016), and progression-free survival (p = 0.010) compared to C allele carriers. Multivariate Cox regression revealed that ATA risk (HR = 14.210, p = 0.000004) and HOTAIR rs920778 (HR = 2.811, p = 0.010) emerged as independent prognostic factors in PTC. A novel polymorphism, MIR155HG rs1893650, was negatively correlated with susceptibility to PTC, with TC heterozygotes exerting a protective effect (OR = 0.268, p = 0.0001). These results suggest that the polymorphisms HOTAIR rs920778 and MIR155HG rs1893650 could be potential prognostic and risk biomarkers in papillary thyroid carcinomas.

Correlation of the OCT Double-Layer Sign with Type 1 Non-Exudative Neovascularization on OCT-A in Age-Related Macular Degeneration.

Background and Objectives: Early diagnosis of the exudative form of age-related macular degeneration (AMD) is very important for a timely first treatment, which is directly related to the preservation of functional visual acuity over a long period. The goal of this paper was to examine the correlation between the double-layer sign (DLS) and the presence of non-exudative macular neovascularization (MNV). Materials and Methods: Our research included 60 patients with AMD, exudative in one eye and non-exudative in the other eye. We analyzed only the non-exudative form using optical coherence tomography (OCT) and optical coherence tomography angiography (OCT-A). The patients were classified into three groups, depending on the duration of the disease (<2 years, 2 to 5 years, >5 years). The onset of the disease was deemed the moment of establishing a diagnosis of exudative AMD in one eye. We defined the presence or absence of a DLS using OCT and the presence of non-exudative MNV using OCT-A, both on 3 × 3 mm and 6 × 6 mm sections. DLS was used as a projection biomarker for non-exudative MNV, with the aim of establishing a rapid diagnosis and achieving early treatment of the disease. Results: We found that there was a statistically significant correlation between the DLS diagnosed using OCT and non-exudative MNV diagnosed by OCT-A for both 3 × 3 mm (p < 0.001) and 6 × 6 mm (p < 0.001) imaging. There was a statistically significant difference between the frequencies of both DLS and MNV in Groups I and III on both 3 × 3 and 6 × 6 mm imaging. A statistically significant difference was also noted in the frequencies of DLS and MNV on 6 × 6 mm imaging, but not on 3 × 3 mm imaging, between Groups I and II. No differences were found between the frequencies of DLS and MNV between Groups II and III. Conclusions: The DLS on OCT can be used as a projection biomarker to assess the presence of a non-exudative MNV.

Examining the Effects of Hyperbaric Oxygen Therapy on the Cardiovascular System and Oxidative Stress in Insulin-Treated and Non-Treated Diabetic Rats.

BACKGROUND: This study explored the effects of hyperbaric oxygen therapy (HBOT) on the cardiovascular system and oxidative stress in streptozotocin-induced diabetic rats. Wistar albino rats were divided into four groups: DM group (diabetic rats), DM+HBOT group (diabetic rats exposed to HBOT for 1 h daily, five days a week, at 2.8 atmosphere absolute (ATA) with 100% oxygen for two weeks), DM+INS group (diabetic rats treated with neutral protamine hagedorn (NPH) insulin at a dosage of 3-5 U/day), and DM+HBOT+INS group (diabetic rats treated with both NPH insulin and HBOT for two weeks). METHODS: Evaluations included glycemic control, oxidative stress parameters, and cardiac function measurements. RESULTS: NPH insulin treatment reduced blood glucose levels, although normoglycemia was not achieved. The DM+HBOT+INS group demonstrated the lowest pro-oxidative marker levels. NPH insulin treatment improved cardiac function, and combination therapy effectively restored cardiac function in diabetic animals. CONCLUSIONS: NPH insulin treatment reduced hyperglycemia and improved cardiac function in diabetic rats. The combined approach of NPH insulin and HBOT resulted in decreased pro-oxidative markers. These findings provide valuable insights for managing cardiovascular complications and oxidative stress in diabetes.

The еffects of low mineral content water on microbiota, metabolic, and oxidative stress parameters in patients with type 2 diabetes mellitus.

Although scientific evidence has shown that natural mineral waters have potential beneficial metabolic effects, there is still very scarce data on their influence on type 2 diabetes mellitus (T2DM). The study was designed to investigate the effects of low mineral water from the "Sneznik-1/79″ source in Serbia on microbiota, metabolic, and oxidative stress parameters in patients with T2DM. In total, 60 patients with confirmed T2DM were included in the study, and they consumed "Sneznik-1/79″ water for 28 days. To examine the positive effects of "Sneznik-1/79″ water, we compared the results before and after the four weeks of "Sneznik-1/79″ water intake. Standard biochemical analyses were carried out, such as glucose level, lipid profile, and stool tests. The blood samples were collected to evaluate the effects of "Sneznik-1/79″ water on the redox status. At the end of the monitoring period, the total cholesterol concentration significantly dropped compared to the initial value. A significant improvement in intestinal peristalsis was observed, which was reflected in the fact that after four weeks, all patients established regular, daily bowel movements. Moreover, consumption of "Sneznik-1/79″ water eliminated the appearance of dysbiosis in 50% of patients. Additionally, the antioxidant capacity was improved by increasing the concentration of superoxide dismutase and reduced glutathione. The result of our study pointed out that the intake of "Sneznik-1/79″ water could be a promising adjuvant therapy for improving intestinal peristalsis as well as reducing the appearance of dysbiosis in T2DM patients.

The acute effects of different spironolactone doses on cardiac function in streptozotocin-induced diabetic rats.

Currently, cardiovascular diseases are the leading cause of global mortality, while diabetes mellitus remains an important cause of cardiovascular morbidity. A recent study showed that patients with diabetes mellitus treated with mineralocorticoid receptor antagonists have improved coronary microvascular function, leading to improved diastolic dysfunction. In this study, we evaluated the influence of acute administration of spironolactone on myocardial function in rats with streptozotocin-induced diabetes mellitus, with special emphasis on cardiodynamic parameters in diabetic rat hearts. The present study was carried out on 40 adult male Wistar albino rats (8 weeks old). Rats were randomly divided into 4 groups (10 animals per group): healthy rats treated with 0.1 µmol/L of spironolactone, diabetic rats treated with 0.1 µmol/L of spironolactone, healthy rats treated with 3 µmol/L of spironolactone, and diabetic rats treated with 3 µmol/L of spironolactone. Different, dose-dependent, acute responses of spironolactone treatment on isolated, working diabetic and healthy rat heart were observed in our study. In healthy rats, better systolic function was achieved with higher spironolactone dose, while in diabetic rats, similar effects of low and high spironolactone dose were observed.

The effects of zofenopril on cardiac function and pro-oxidative parameters in the streptozotocin-induced diabetic rat heart.

Diabetes mellitus is a chronic condition that continues to increase in both incidence and prevalence. Renin-Angiotensin-Aldosterone System is one of the main modulators of chronic hyperglycaemia and, thus, its influence on tissues. Hyperglycaemia-induced oxidative stress is an important factor in diabetic cardiomyopathy. The present study was carried out on 24 adult male Wistar albino rats (8-week-old and with body masses of 190 ± 10 g). We evaluated the influence of acute administration of zofenopril on ex vivo myocardial function from rats with streptozotocin-induced diabetes mellitus, with a special emphasis on cardiodynamic and oxidative stress parameters in diabetic rat hearts. Rats were divided randomly into two groups (12 animals per group): control non-diabetic animals (C) were healthy rats perfused with 1.5 µM of zofenopril, and STZ-treated diabetic animals (DM) were diabetic animals perfused with 1.5 µM of zofenopril 4 weeks after the induction of diabetes. Our results demonstrated that diabetic rats are characterized by a depressed cardiac performance and that oxidative markers are related to alterations in cardiac function in rats with 4 weeks of STZ-induced diabetes. Additionally, the use of zofenopril as a monotherapy slightly diminished cardiac damage induced by chronic hyperglycaemia. However, long-term follow-up intervention trials are necessary to fully demonstrate the benefit of zofenopril in this context. A challenge for future investigations will be to identify the effects of chronic administration or combination therapy with angiotensin-converting enzyme inhibitors in various models of diabetes.

Catheter-associated urinary tract infection in a surgical intensive care unit.

BACKGROUND/AIM: Because patients in intensive care units usully have an urinary catheter, the risk of urinary tract infection for these patients is higher than in other patients. The aim of this study was to identify risk factors and causative microrganisms in patients with catheter-associated urinary tract infection (CAUTI) in the Surgical Intensive Care Unit (SICU) during a 6-year period. METHODS: All data were collected during prospective surveillance conducted from 2006 to 2011 in the SICU, Military Medical Academy, Belgrade, Serbia. This case control study was performed in patients with nosocomial infections recorded during surveillance. The cases with CAUTIs were identified using the definition of the Center for Disease Control and Prevention. The control group consisted of patients with other nosocomial infections who did not fulfill criteria for CAUTIs according to case definition. Results. We surveyed 1,369 patients representing 13,761 patient days. There were a total of 226 patients with nosocomial infections in the SICU. Of these patients, 64 had CAUTIs as defined in this study, and 162 met the criteria for the control group. Multivariate logistic regression analysis identified two risk factors independently associated to CAUTIs: the duration of having an indwelling catheter (OR = 1.014; 95% CI 1.005-1.024; p = 0.003) and female gender (OR = 2.377; 95% CI 1.278-4.421; p = 0.006). Overall 71 pathogens were isolated from the urine culture of 64 patients with CAUTIs. Candida spp. (28.2%), Pseudomonas aeruginosa (18.3%) and Klebsiella spp. (15.5%) were the most frequently isolated microorganisms. CONCLUSIONS: The risk factors and causative microrganisms considering CAUTIs in the SICU must be considered in of planning CAUTIs prevention in this setting.

Role of decreased production of interleukin-10 and interferon-gamma in spontaneous apoptosis of B-chronic lymphocytic leukemia lymphocytes in vitro.

BACKGROUND AND AIMS: B-chronic lymphocytic leukemia (B-CLL) is characterized by the progressive accumulation of small B lymphocytes that do not undergo apoptosis due to an underlying defect. One potential mechanism of defective apoptosis may be irregular cytokine production. The goal of our investigation was to determine in vitro production of relevant cytokines by lymphocytes of B-CLL patients. METHODS: Thirty untreated (stage A) B-CLL patients, as well as 20 stage B and C patients and 30 healthy volunteers as a control group were examined. Interleukin-4 (IL-4), interferon-gamma (IFN-gamma), interleukin-10 (IL-10) and tumor necrosis factor-alpha (TNF-alpha) were measured by enzyme-linked immunosorbent assay (ELISA) in supernatants of lymphocyte cultures of all three investigated groups. The method applied for detecting apoptosis was fluorescence microscopic analysis using acridine orange/ethidium bromide (AO/EB) double staining. RESULTS: Investigation showed that in vitro lymphocyte production of IL-10 and IFN-gamma were significantly decreased in B-CLL patients, whereas there were no statistically significantly differences of IL-4 and TNF-alpha production among the tested groups. Compared with the spontaneous apoptosis observed in control subjects' lymphocytes, B-CLL lymphocytes showed increased percentages of apoptotic cells after incubation for 24h. Interestingly, increased spontaneous apoptosis of B-CLL lymphocytes was followed by decreased IL-10 and IFN-gamma production. Stage of disease did not influence B-CLL lymphocyte spontaneous apoptosis in vitro. CONCLUSIONS: These changes in cytokine production in cultures of B-CLL lymphocytes may be one of the potential mechanisms in the pathogenesis of abnormal apoptosis.

Clinical evaluation of the simultaneous determination of CA 15-3, CA 125 and sHER2 in breast cancer.

OBJECTIVE: We investigated serum levels of CA 15-3, sHER2 and CA 125, and their usefulness in the detection of metastatic disease in breast cancer patients. METHODS: The levels of CA 15-3, sHER2 and CA 125 tumour markers were determined in 60 patients, 40 with localized and 20 with metastatic breast carcinoma. The control group consisted of 10 healthy women. RESULTS: We found that, at the time of diagnosis, serum levels of all three tumour markers were elevated in patients with distant metastases, but of minute importance in the detection of any breast cancer. When the data for the individual markers were combined the overall sensitivity of metastases detection with all three markers improved. In this regard, 90% of patients with distant metastases had an increase in serum level of at least one of tested tumour markers. Similar results were obtained using receiver operating characteristic curve (ROC). Moreover, using ROC we defined cut-off values for metastasis detection for each of the tested markers. CONCLUSION: Our findings indicate that measurement of CA 15-3 serum values in conjunction with sHER2 and CA 15-3 can increase sensitivity in metastasis detection.

Analysis of cycloheximide-induced apoptosis in human leukocytes: fluorescence microscopy using annexin V/propidium iodide versus acridin orange/ethidium bromide.

Apoptosis is a highly regulated and programmed cell breakdown process characterized by numerous changes. Since it is implicated in many pathological as well as physiological processes, it is vital to have reliable methods for detecting cell death. In this study, we compared several methods for detecting apoptosis and necrosis in human leukocytes. Apoptosis was induced either by incubating the cells with various doses of cycloheximide (CHX) or by 312 nm UVB irradiation. The methods used for detecting apoptosis were light microscopy (May Grunwald-Giemsa and trypan blue staining), fluorescence microscopy (acridin orange/ethidium bromide and annexin V/propidium iodide staining) and agarose gel electrophoresis of fragmented genomic DNA. Our study showed that CHX-induced apoptosis in cultured peripheral blood mononuclear cells but had no effect on apoptosis in polymorphonuclear cells, so its effect depends on cell type. Evaluation and comparison of the methods for detecting apoptosis showed the following. A Giemsa-stained cytospin allows the main morphological characteristics of necrotic and apoptotic death to be recognized. Trypan blue staining, widely used for estimating cell viability, is valueless for detecting apoptosis. Both fluorescence methods provided reliable and reproducible results and distinguished clearly between subpopulations of apoptotic cells, and were closely intercorrelated. Although applicable to a wide spectrum of cell types, agar electrophoresis of extracted DNA cannot be applied to all cell types and apoptotic conditions. Generally, microscopic examination of acridin orange/ethidium bromide stained cells can be recommended as the most reliable of the methods tested.