Petar Canovic

Induction of mitochondrial apoptotic pathway by raloxifene and estrogen in human endometrial stromal ThESC cell line

Introduction Endometrial hyperplasia is a condition that occurs as a result of hormonal imbalance between estrogen and progesterone. Morphological disturbance of endometrial cells occurs consequently leading towards endometrial cancer. In therapy of endometrial hyperplasia SERMs are used to supress effects of locally high estrogen level in uterus. There is strong evidence suggesting that estrogen could be involved in cell death – apoptosis. There are no experimental data demstrating the direct apoptotic effect of both raloxifene and estrogen on the ThESC cell line. The aim of our study wa sto investigate both cytotoxic and apototic mechanism of raloxifene and estrogen – induced death in the ThESC cell line. Material and methods In order to determine their cytotoxic and apoptotic effects, various doses of raloxifene and estrogen were applied to the ThESC cell line for 24 h. After the treatment MTT assay, FACS analysis and immunofluoroscence method were conducted. Results The results of this study for the first time demonstrated the cytotoxic and apoptotic effects of raloxifene and estrogen on human endometrial stromal cell line suggesting the involvement of the inner, mitochondrial apoptotic pathway. Conclusions Our results demonstrated apoptotic effects of investigated drugs in the ThESC cell line through increasing the Bax/Bcl-2 ratio and activation of caspase 3.

Older Hypertensive Patients’ Adherence to Healthy Lifestyle Behaviors

Abstract Non-pharmacological treatment including diet, body weight reduction, smoking cessation and physical activity, is very important part of hypertension treatment. The objective of this study was to investigate the adherence to healthy lifestyle behavior in the representative sample of the older hypertensive patients, and to investigate factors associated with adherence in the studied older population. The study was conducted on random sample of 362 long term hypertensive (> five years) patients older than 65 years of age, at Health Care Center of Kragujevac. Adherence was assessed using the structured questionnaire for the analysis of the implementation of both hypertension and diabetes guidelines in the primary care. Both bivariate and multivariate analyses were conducted. Nearly 35% of examined patients were highly adherent; they exercised regularly, avoided smoking for at least five years and consumed special healthy diet prescribed for hypertension. Another 35.6% of the cases reported exercising regularly, 39.5% followed the recommended diet for the hypertension, while 23.4% of the patients have still consumed cigarettes. Multivariate logistic regression demonstrated that received counseling on healthy lifestyle behaviors by physicians and lack of education predicted high adherence to healthy lifestyle behavior. In order to improve adherence of elderly hypertensive patients to healthy lifestyle, strengthening patient-physician relationships through efforts to enhance communication may be a promising strategy to enhance patients’ engagement in healthy lifestyle behaviors for hypertension. Such an improvement could be achieved through the education of both the physicians and patients.

Acute Kidney Damage: Definition, Classification and Optimal Time of Hemodialysis

Abstract Acute damage to the kidney is a serious complication in patients in intensive care units. The causes of acute kidney damage in these patients may be prerenal, renal and postrenal. Sepsis is the most common cause of the development of acute kidney damage in intensive care units. For the definition and classification of acute kidney damage in clinical practice, the RIFLE, AKIN and KDIGO classifications are used. There is a complex link between acute kidney damage and other organs. Acute kidney damage is induced by complex pathophysiological mechanisms that cause acute damage and functional disorders of the heart (acute heart failure, acute coronary syndrome and cardiac arrhythmias), brain (whole body cramps, ischaemic stroke and coma), lung (acute damage to the lung and acute respiratory distress syndrome) and liver (hypoxic hepatitis and acute hepatic insufficiency). New biomarkers, colour Doppler ultrasound diagnosis and kidney biopsy have significant roles in the diagnosis of acute kidney damage. Prevention of the development of acute kidney damage in intensive care units includes maintaining an adequate haemodynamic status in patients and avoiding nephrotoxic drugs and agents (radiocontrast agents). The complications of acute kidney damage (hyperkalaemia, metabolic acidosis, hypervolaemia and azotaemia) are treated with medications, intravenous solutions, and therapies for renal function replacement. Absolute indications for acute haemodialysis include resistant hyperkalaemia, severe metabolic acidosis, resistant hypervolaemia and complications of high azotaemia. In the absence of an absolute indication, dialysis is indicated for patients in intensive care units at stage 3 of the AKIN/KDIGO classification and in some patients with stage 2. Intermittent haemodialysis is applied for haemodynamically stable patients with severe hyperkalaemia and hypervolaemia. In patients who are haemodynamically unstable and have liver insufficiency or brain damage, continuous modalities of treatment for renal replacement are indicated.

Enhanced cytotoxicity and apoptosis by raloxifene in combination with estrogen and methotrexate in human endometrial stromal cells

Endometrial hyperplasia is a condition that may lead to the development of endometrial carcinoma. Initially, changes of the endometrium are caused by the estrogens hyperstimulation that may lead to the development of an irregular bleeding and the infertility problems. Therapy of endometrial hyperplasia is limited to medical and surgical approaches. During the past decade, the new types of drugs were developed for the treatment of the endometrial hyperplasia. Here, for the first time, we investigated the cytotoxic effects of the various combinations of estrogen, raloxifene, and methotrexate in human ThESC cell line as a possible potential treatment of the endometrial hyperplasia. Our aim was to investigate and to determine the most efficient combination of investigated drugs in ThESC cells during 24‐hr period using MTT assay, FACS analysis, and immunofluorescence staining. Our results demonstrated that the combination of raloxifene with methotrexate efficiently induced both the cytotoxicity and apoptosis in ThESC cells when compared to their single effect, as well as to the effect of combined treatment of raloxifene with estrogen. The application of the low doses of methotrexate combined with raloxifene offers all advantages of a potential beneficial antitumor match in cancer chemoprevention and therapy.

Tumor Necrosis Factor- Alpha as Differential Diagnostic Marker for Patients with Fever of Unknown Origin

Abstract Febrile conditions of unidentified origin are still unknown in modern medicine despite the development of diagnostic procedures. There are various agents of long-term temperature encompassing numerous infectious or non-infectious diseases. The aim of this study was to determine if there was a statistically significant difference in the values of proinflammatory cytokines (IL-1, TNFα, IL-6) in patients who meet the criteria for febrile conditions of unidentified origin, between the group of infectious, malignant, rheumatic, “other” diseases and undiagnosed patients. The study was conducted in the Immunology laboratory of the Center for Molecular Medicine and Stem Cells Research of the Faculty of Medical Sciences in Kragujevac. Blood samples were taken from patients tested at the Clinic for Infectious Diseases, of the Clinical Center of Kragujevac, in the period from 2014 to 2016. The study included 70 patients. The measured values of the level of TNFα showed significantly higher values in a group of malignant diseases than in the group of infectious diseases, while the values of IL-1 and IL-6 did not show statistical significance. TNFα can improve diagnosing in case of patients with an unknown febrile condition, which can shorten the length of the hospital stay and reduce the volume of performance of diagnostic procedures.

The Analysis of Risk Factors and Clinical-Demographic Characteristics of Patients with Clostridium Dificille Infection as Well as The Outcome of Their Treatment

Abstract Pseudomembranous colitis is a frequent nosocomial infection associated with significant morbidity and mortality. Clostridium difficile infection incidence most frequently increases due to unreasonable antibiotic use and the appearance of new hypervirulent bacterial strains, which leads to prolonged hospitalization and an increase in the total cost of hospital treatment. This is a retrospective design study conducted at Clinical Centre Kragujevac from January to December 2014. The patient data were obtained from the protocol of the Virological Laboratory and from medical documentation. All statistical analyses were performed using the computer program SPSS. The descriptive statistical data are expressed as percentage values. Continuous variables are expressed as the arithmetic mean with the standard deviation. Clostridium difficile infection occurred more frequently with elderly patients (123 patients were over 65 years old). Out of 154 patients on antibiotic treatment, 110 patients were treated with a combination of two or more antibiotics from different pharmacological groups. The most represented antibiotics were from the cephalosporin (71.4%) and quinolone (46.3%) groups. A total of 85.8% of the patients used proton pump inhibitors and H2 blockers. Our results describe the clinical and demographic characteristics of patients with diagnosed Clostridium difficile infection. The most prevalent characteristics (age, antibiotic therapy, PPI and H2 blocker use), which other researchers have also mentioned as risk factors, were present in our study as well.

Analysis Of Clinical, Haematological And Biochemical Parameters In Patients With Infectious Mononucleosis

Abstract Primary infection with Epstein-Barr virus (EBV) usually occurs in early childhood and often does not present clinical symptoms. More than 90% of adults are infected with this virus. A primary infection that occurs in adolescence or adulthood is usually clinically presented as infectious mononucleosis with a triad of symptoms: fever, lymphadenopathy and pharyngitis. Our retrospective study included 51 patients with a median age of 17 (9-23) years and serologically confirmed infectious mononucleosis. All patients with infectious mononucleosis were treated at the Clinic for Infectious Diseases at the Clinical Center in Kragujevac during 2013. We analysed the clinical, haematological and laboratory parameters of patients. The aspartate-aminotransferase levels were increased in 40 patients, with a mean value of 116.24 (±93.22); the alanine-aminotransferase levels were increased in 44 patients, with a mean value of 189.24 (±196.69). Lymphadenopathy was the most common clinical feature upon admission in 49 patients (96%); 38 patients (74.5%) had splenomegaly, and 20 (39%) had hepatomegaly. Twenty-six patients (51%) had leukocytosis with lymphocytosis, while 15 (75%) of the 20 who had a normal leukocyte count also had lymphocytosis. In the present study, we updated the clinical, haematological and laboratory parameters, which may lead to the establishment of an accurate diagnosis and promote further treatment of the patients.