Clinical use of an analysis of oxidative stress and IL-6 as the promoters of diabetic polyneuropathy.

Aim To investigate interleukin 6 (IL-6) values depending on duration of diabetes mellitus (DM) and evaluate possible correlation with diabetic polyneuropathy. Methods The research study included 90 patients with DM divided into three groups (30 patients each) according to the duration of DM: group A - patients who had DM for less than 10 years, group B - duration of DM was 10 to 20 years, and group C - patients with DM over 20 years. Control group (K) included 30 healthy participants. Results IL-6 was significantly higher in the healthy control group, 180.318 pg/mL±94.18, than in group A, 47.23pg/ml±34.8, group B, 43.31pg/ml±33.17, and group C, 70.39 pg/ml±59.26 (p=0.0001). All groups had significantly different values of IL-6 between each other (p=0.0001). Level of IL-6 was in correlation with diabetic polyneuropathy in the group A (the youngest participants) (p=0.0001). In other groups there was no significant correlation between IL-6 and diabetic polyneuropathy. Conclusion The level of IL-6 was in correlation with neuropathy among younger patients. A higher level of IL-6 in the control group than in diabetic groups is a sign of stronger inflammatory response among younger and healthy people than in patients with DM.

Serum Uric Acid Concentration in Patients with Cerebrovascular Disease (Ischemic Stroke and Vascular Dementia).

INTRODUCTION: Significance of serum uric acid (UA) in cerebrovascular disease still remains controversial. UA is most abundant natural antioxidant in human plasma. Its antioxidant properties might protect against free radical damage, thereby reducing the risk of oxidative stress-related cognitive impairment and dementia. AIM: In our investigation, we determine the level of UA in 100 male patients diagnosed with the first ischemic brain stroke (blood samples were collected during the acute phase and post-acute phase), 100 male patients diagnosed with vascular dementia and 100 male healthy volunteers (control group). METHODS: UA was determined using DIMENSION LxR automatic analyzer. Measurement of UA concentration was based on an enzymatic method (range 208-428 µmol/L). RESULTS: The prevalence of hyperuricemia among ischemic stroke and vascular dementia patients was 30% and 8%, respectively. Serum UA concentration was higher 7 and 14 days after the stroke compared to the acute phase (24-48 hours after hospitalization) and these concentrations were significantly higher than those measured in the control group. UA levels measured at 24-48 hours after the first symptoms of ischemic stroke were strongly correlated with those measured after 7 days of treatment (r = 0.79, p = 0.001) or after 14 days (r = 0.839, p = 0.0049). No significant differences were found between ischemic stroke and vascular dementia groups. CONCLUSION: UA concentrations were higher in ischemic stroke and vascular dementia groups than in controls. UA increase may reflect vascular atherosclerosis and tissue hypoxia. UA monitoring in patients with cerebrovascular disease is essential, because UA is more harmful than protective.

The Estimation of Glomerular Filtration Rate Based on the Serum Cystatin C and Creatinine Values.

BACKGROUND: Renal insufficiency is one of the most serious renal diseases. Early diagnosis plays an essential role. Serum creatinine as a marker has had priority so far. The use of cystatin C for estimation of glomerular filtration rate has been recommended recently. There is a set of formulae which consolidates serum values of both creatinine and cystatin C. The study focuses on different formulae and their estimation for the purpose of better, faster, and more accurate diagnosis of renal insufficiency. METHODS: The sample consists of 75 examinees which are divided into three groups. The first group is made of patients with the glomerular filtration rate (GFR) 90 - 120 mL/minute/1.73m². The second group consists of patients with GFR 60 - 89 mL/minute/1.73m² and the third with GFR 30 - 59 mL/minute/1.73m². The exclusion criteria are the following: children and adolescents under the age of 20, pregnant women, patients on corticosteroid therapy and blockers of the distal tubular creatinine secretion. The five equations (GFR1-5) used in this research are: a Cockcroft-Gault equation - GFR1; a MDRD equation - GFR2; CKD-EPI - an equation based on cystatin C - GFR3; CKD-EPI - an equation based on cystatin C, adjusted according to the gender and age - GFR4; and CKDEPI - a combined equation based on cystatin C and creatitine, adjusted according to the age, gender, and race - GFR5. Upon acquiring the results, it is followed up with the statistical data analysis followed by graphs and tables. RESULTS: After data analysis, it is established that data distribution does not show normal distribution in each case, which leads to the use of nonparametric statistics. Depending on the stage of kidney injury there are different results regarding the difference in statistics of the used formulae. The highest sensitivity is recorded with the formulae GFR4 and GFR5. Then the increase in cystatin C levels increases sensitivity with the formulae GFR3 and GFR4. CONCLUSIONS: As a result of this study, it is to be established that the formula of choice is the GFR3 - CKD-EPI formula based on serum cystatin C values, without adjustments. Its sensitivity, specificity, price, and feasibility are to be observed as parameters. Besides that, the increase in serum cystatin levels leads to the increase of sensitivity of the GFR3 formula, which could be an additional factor in the selection of formulae.

Vascular endothelial growth factor receptor-1 expression in breast cancer and its correlation to vascular endothelial growth factor a.

VEGF-A is the most potent angiogenic factor in tumour angiogenesis. Its effects are mediated via two receptors VEGFR-1 and VEGFR-2. Primary aim of our study was to examine the expression of VEGFR-1 in breast cancer and its correlation to VEGF expression, lymph node status, tumour size, histological grade, and hormone receptor status. To examine the VEGFR-1 and VEGF expressions in tumour and surrounding tissue of 51 breast cancer patients, and in healthy breast tissue of 30 benign breast diseases patients, we used three-step immunohistochemical staining. VEGFR-1 and VEGF expressions were significantly increased in breast cancer tumour in relation to surrounding tissue (P < 0.01), and the VEGF expression was significantly increased in lymph node positive breast cancer patients (P < 0.01). VEGFR-1 and VEGF expressions were significantly higher in breast cancer tumour compared with healthy breast tissue (P < 0.01). Significant correlation between VEGF and VEGFR-1 expressions was found (P < 0.05). No significant correlations between VEGF and VEGFR-1 expressions and tumour size, histological grade, and hormone receptor status were found. Increased expression of VEGFR-1 and VEGF in breast cancer tumour and significant correlation between these proteins suggest the possible role of VEGF/VEGFR-1 signalization in breast cancer development, although VEGFR-1 potential prognostic value was not confirmed.

Idiosyncratic adverse reactions of most frequent drug combinations longterm use among hospitalized patients with polypharmacy.

GOAL: Inappropriate prescribing of a multiple therapeutic agents to patients with chronic conditions is very common in everyday practice. Adverse drug reactions (ADRs) are still considered as one of the main problems of drug therapy. We investigated idiosyncratic symptoms and signs of adverse drug reactions (ADRs) of the most frequent used combination of drugs among hospitalized patients prescribed polypharmacy. METHODOLOGY: A cross sectional study (design) was performed in Pharmacies "Eufarm Edal" in Tuzla from 2010 to 2011. Polypharmacy was defined as using > or =4 drugs. The total study sample of 166 examiners was interviewed with a questionnaire about ADRs which was developed special for study. Linear regression analyses was used to evaluate predictors of idiosyncratic signs of adverse drug reactions of the most prevalent drug combinations; using length of drugs in cases polypharmacy more than 6 months as independent variable. Age, sex, index of cumulative morbidity, drug number in polypharmacy, type of drug combination related pharmacological effects, type of hospital clinics were used as possible confounders. RESULTS: The most common exposures to various drug combinations were: medication for high blood pressure and heart (62%), psychotropic drugs (59%), antacids (47%) and antibiotics (46%) among hospitalized patients with polypharmacy. Our results indicated that from 9.6% to 90.4% of hospitalized patients with polypharmacy had at least one suspicious long-term idiosyncratic drug combination use symptoms. The ADRs prevalence often used psychotropic drug combination was initiated suspected idiosyncratic adverse reactions: confusion, depression, anxiety, decreased libido and insomnia. Linear regression analyses also showed that it remains a very strange, and negative idiosyncratic and lacking therapeutic effects of use of antacids in conditions of polypharmacy. CONCLUSION: The toxicity of some drug combinations may sometimes be synergistic and be greater than the sum of the risks of toxicity of either agent used alone. In order to recognize and to prevent ADRs (including drug interactions), good communication between pharmacist and patient and/or physicians and patient is crucial, and prescribers should develop an effective therapeutic partnership with the patient and with fellow health professionals.

Prevalence of polypharmacy and drug interaction among hospitalized patients: opportunities and responsabilities in pharmaceutical care.

GOAL: Polypharmacy and drug-related problems (DRPs) have been shown to prevail in hospitalized patients. We evaluated the prevalence of polypharmacy; and investigated relationship between polypharmacy and: symptoms of DRPs, number of drugs and OTC, index of cumulative morbidity, length of exposure to polypharmacy and the number of days of hospital stay among hospitalized patients. METHODOLOGY: A study was performed in Pharmacies "Eufarm Edal" Tuzla from 2010 to 2011. Polypharmacy was defined as using ≥ 3 drugs. The total study sample of 226 examiners were interviewed with special constructed questionnaires about DRPs. Experimental study group consisted of hospital patients with polypharmacy (n=166) and control group hospital patients without polypharmacy (n=60). Mann-Whitney test was used to test for significant self-reported symptom differences between groups and cross sectional subgroups, t- test and χ(2)- test for age, gender and treatment data in hospital. RESULTS: The prevalence of polypharmacy was 74% among 226 hospitalized patients. The vulnerable age subgroup of hospitalized patients was men and hospitalized patients aged from 46 to 50 years (not geriatric patients). The prevalence of index of cumulative morbidity was 65%. The most common exposures varied by patient age and by hospital type, with various antibiotics, antidepressants, analgesics, sedatives, antihypertensives, flixotide, ranitidine and others. The prevalence of exposure to OTC and self- treatment was 80%. The prevalence of symptoms of drug-related problems were significantly differed among patients of experimental in relationship of control study group patients (P<0.001). CONCLUSION: In addition to helping to resolve the above mentioned issues, the results from this study could provide baseline information quantifying the problem of drug- related problems among hospitalized patients receiving polypharmacy and contribute to the formulation and implementation of risk management strategies for pharmacists and physicians in primary care health.

Interleukin 13 expression in the primary breast cancer tumour tissue.

INTRODUCTION: The aim of this study was to investigate the presence and the expression levels of the interleukin 13 (I1-13) in the primary breast cancer tumour tissue in relation to the unchanged breast tissue in the same patients and to the breast tissue in the patients with benign breast disease, and to investigate the correlation between the IL-13 expression levels and the pathohistological factors, and between IL-13 expression and estrogens and progesterone receptor status. MATERIALS AND METHODS: 50 patients with invasive ductal breast cancer and 20 patients with benign breast diseases were included in this prospective case-control study. The three-step immunohistochemical staining was used for testing the levels of IL-13 expression and hormone receptor status. RESULTS: IL-13 was present in breast cancer tumour tissue, and in the surrounding unchanged tissue in the same patients, and in breast tissue in patients with benign breast disease. The expression of IL-13 was significantly higher in breast cancer tumour compared with surrounding tissue (P < 0.05) of the same, lymph node-positive patients. In addition, IL-13 expression was significantly higher in breast cancer tumour compared with breast tissue in patients with benign breast diseases (P < 0.01). There was significant correlation between IL-13 expression and tumour size in patients with lymph node-negative breast cancer (r = 0.405, P = 0.050). There was no significant correlation between IL-13 expression and the other pathohistological factors, and no significant correlation between IL-13 expression and the lymph node status. CONCLUSION: Obtained results suggest possible involvement of IL-13 in breast carcinogenesis.

Cystatin C in sera of patients with aggressive non-Hodgkin B-cell lymphoma.

AIM: To investigate the cystatin C levels in sera of patients with aggressive non-Hodgkin B-cell lymphoma. METHODS: The levels of cystatin C in sera of lymphoma patients and control group consisted of healthy individuals, were measured by using specific sandwich-type ELISA. For each patient the clinical stage of disease was determined according to Ann Arbor staging system for lymphomas. RESULTS: Our study shows that mean cystatin C serum level in the patients group (1056 +/- 65 ng/mL) was significantly higher when compared with the mean level of the healthy control group (819 +/- 28 ng/mL) (P = 0.001). Mean cystatin C level of the group with clinical stages III and IV (1255 +/- 109 ng/mL) was significantly elevated when compared with the mean level of the group with clinical stages I and II (896 +/- 51 ng/mL) (P = 0.03). CONCLUSION: This finding points out a connection between inhibitor level and aggressive behaviour of lymphoma and could be considered for further strategies of prognosis of the disease.

Cystatin C as a potential marker for relapse in patients with non-Hodgkin B-cell lymphoma.

The concentration of cysteine protease inhibitor cystatin C was determined in sera from 59 patients with non-Hodgkin B-cell lymphoma using ELISA. The sera from 43 age and sex matched healthy blood donors served as controls. Cystatin C was significantly increased in sera of patients without therapy (mean 1136+/-SE 105.7ng/ml, p=0.00001) and with therapy (mean 1073+/-52ng/ml, p=0.001) compared to controls (mean 819+/-28ng/ml). The highest levels were determined in sera of patients with a relapse (mean 1680+/-196ng/ml). By using immunofluorescence staining and confocal microscopy we determined immature dendritic cells as a major population of cystatin C positive cells in affected lymph nodes. Our study reports for the first time that cystatin C is a potential marker for relapse in patients with non-Hodgkin B-cell lymphoma.

[Morphological research of fibrinoid and intervillous space of human term placenta in relation to the age of pregnant woman]. / Morfoloska istrazivanja fibrinoida i intervilusnog prostora humane zrele posteljice u odnosu na zivotnu dob trudnice.

Fibrinoid and intervillous spaces of the mature human placenta were morphologically and quantitatively examined in pregnant women of different ages. The aim of this research was to compare the results and to confirm if there is some quantitative difference between fibrinoid and intervillous spaces of placenta related to the age of pregnant women. The examined group of women were of age between 20 and 45 years, and they were divided into two groups: 1) pregnant women 20-35 year old; 2) pregnant women over 35 years old. The volume density and absolute volume of fibrinoid and intervillous space of placenta were stereologically examined. Volume densities of fibrinoid and intervillous space of placentas in older pregnant women compared to younger is significantly increased. Absolute volumes of fibrinoid and intervillous space of placentas in younger and older pregnant women are not significantly different.

Temptation of academic medicine: second alma mater and "shared employment' concepts as possible way out?

Apparently, in developing and in well-developed societies we are confronted with a crisis of academic medicine in all aspects: health care, teaching, and research. Health care providers in teaching hospitals are under pressure to generate revenues, academic research is pressed to keep pace with institutions devoted solely to research, and teaching is often understood not as privilege and honor but as burden and nuisance. The key problem and the principal cause of the crisis are low interest of the best young graduates to follow an academic career in a world where the benefits and values of the private sector are prevailing. Confronted with these circumstances and the continuous perils of permanent brain-drain, we developed an innovative concept of "shared employment' where two academic institutions (one in a developed and one in a developing country) will collaborate in development and support of fresh talents, building elite academic staff. Most academic exchange programs developed so far have proved to be ineffective and of poor vitality, in spite of loud exclamations, high expectations, and a huge amount of good will involved. In contrast, the suggested cooperation will be based exclusively on mutual interest and clearly defined benefits for all involved parties.

[Aminotransferase activity during convalescence in infectious hepatitis in older adolescents]. / Uticaj prebolovanog infektivnog hepatitisa na aktivnosti aminotransferaza u starijih adolescenata.

The objective of the research were consolidate the correlation the greatest values ALT and AST, and the recovery from hepatitis infection. To the group of 237 adolescents, male sex of 18, 19 and 20 years is measured the activity of ALT and AST. The examinees are questionnaired to the recovery from hepatitis infection. Had to the significant increased values AST, and only mildly increased values in relation to the upper referent values.

[Effect of alcohol on aminotransferase activity in adolescents]. / Uticaj alkohola na aktivnosti aminotransferaza u adolescenata.

The objective of research were consolidate the correlation the greatest values ALT and AST, and consummation alcohol. To the group of 302 adolescents, male sex of 18, 19 and 20 years is measured the activity of ALT and AST. The examines are questionned to the consummation of alcohol. Had to the significant increased values AST, and only mildly increased values in relation to the upper referent values.

[Correlation of body mass index and aminotransferase activity in healthy adolescents]. / Korelacija indeksa tjelesne mase i aktivnosti aminotransferaza u zdravih adolescenata.

The objective of this study to consolidate the correlation between BMI with the values of ALT and AST of the healthy adolescents in age of 18, 19 and 20 years. To the group of 237 examinees of the healthy adolescents is determined BMI, and is measured the activity of ALT and AST. In the sample of the examinees we had 22% examinees had the increased BMI > or = 25. The examinees who had the increased BMI (> or = 25), had the significant increased values ALT, and only mildly increased values AST in relation to the upper referent values. On the basis of the simple linear regression was confirmed the positive correlation between the of body mass indexes and activities ALT i AST.

Isolation and immunochemical characterization of human cystitis C.

Cystatin C is a natural inhibitor of the cysteine proteinases papain, and mammalian lysosomal cathepsins B, H, L and S. This protein is thought to serve an important physiological role as a local regulator of enzyme activity. The changes of levels of cystatin C in extracellular fluids have shown themselves having potential clinical importance. We have purified cystatin C from urine of patients with chronic renal failure by procedure using affinity chromatography on CM-papain Sepharose, gel filtration on Sephacryl S-200, and ion exchange chromatography on CM-cellulose. After isolation we obtained three inhibitory peaks (pI's from 7.8 to 9.2) which represent isoforms of the same protein. These isoforms are immunologically identical and differ in N-terminal sequence of the molecule. The form with pI 9.2 represents the intact inhibitor form, whereas the form with pI 7.8 is shortened for 8 amino-acid residues at N-terminal end. Purified cystatin C pI 9.2 was used for immunization of rabbits. Polyclonal antibodies, produced in rabbits, were isolated from rabbit sera by affinity chromatography on Protein A Sepharose. Enzyme immunoassay (ELISA) for cystatin C is developed on the basis of purified antibodies. Using ELISA test we determined amount of cystatin C in urine and serum samples of patients with chronic renal failure. The concentration of the inhibitor in the urine of these patients was approximately 100-fold more than in normal urine. In the serum from the same patients we found concentrations of cystatin C to be five times higher in comparison with the serum of healthy individuals.

[New findings on the diagnosis and therapy of prion diseases]. / Nova saznanja o mogucnostima dijagnostike i terapije prionskih bolesti.

Spongiform encephalopathies are the fatal diseases, that affect the brain tissue of mammals. They are caused by a conformational changed prion protein. There is no adequate diagnostic test for in vivo identification of prion protein. Disease can be diagnosed only by clinical sings and EEG in new variant of Creutzfeldt-Jakob disease. Post mortem, histopathological examination of brain tissue reveals spongiform changes and immunohistochemistry detects disease-related prion protein. Appropriate diagnostic in vivo tests are not developed yet; therefore extensive researches are ongoing aimed to introduce such methods. This review describes a few promising experimental methods, which may develop into diagnostic tests in the future: detection of prions in urine samples, PMCA (protein misfolding cyclic amplification), DATAS (differential analysis of transcripts with alternative splicing), SELEX (in vitro selection), detection of prions in tonsils and detection of copper and manganese dysbalance in tissues. Current therapy strategy is based on testing of some known drugs (quinacrine, chlorpromazine), and antioxidant and antibody treatments. The detection of NSE (neuron-specific enolase) and cholesterol in meat products reveals the presence of brain and spinal cord tissue. The spreading of spongiform encephalopathies can be diminished by utilising the adequate in vivo diagnostic tests, effective therapy strategy and preventive steps.