Ismet Tahirovic

Antioxidant capacity of the neurohormone melatonin

Summary.The aim of this study was to elucidate the antioxidant behaviour of melatonin (M) and determine its activity–structure relationship. M or 5-metoxy-N acetyltriptamine is a neurohormone secreted by the pineal gland, which plays a proven role in maintaining sleep-wake rhythms. The antioxidant capacity of M was analysed using the oxygen radical absorbance capacity (ORAC) assay. Furthermore, spectral measurements for aerobic photolytic reaction of neutral red (NR) and degree of inhibition of photolysis with M, glutathione (GSH), ascorbic acid (AA) and vitamin E analogue Trolox were studied at room temperature 25°C, using visible (VIS) and ultra-violet (UV) radiations. In the ORAC assay 2,2-azobis (2-amidino-propane)dihydrochloride (AAPH) a peroxyl radical generator, ROO°; H2O2–Cu2+, mainly a hydroxyl radical generator, °OH; and Cu2+ a transition metal were used. Although some studies indicated that M is a powerful antioxidant, no one has compared its antioxidant capacities with GSH, E-vitamin and AA, using three free radical (FR) generators in an assay which utilizes an area-under curve technique and thus combines both inhibition time and inhibition degree of FR action by an antioxidant into a single quantity. In the current study, we used ORAC assay with three FR generators. The assay is based on propensity of the fluorescence emitted by the protein β-phycoerythrin (β-PE) from porphyridium cruentum to be quenched when exposed to FR action. M in our experiments acted as a universal antioxidant against ROO° and °OH radicals. Also, M served as an antioxidant in the presence of Cu2+. M, which is a lipid-soluble compound, was a twice more powerful antioxidant than vitamin E, and four times than AA or GSH. Furthermore, M inhibited aerobic photolysis of NR photoinduced with VIS and UV rays faster and more effectively, than AA, GSH or vitamin E. AA with NR, under aerobic conditions during irradiation with VIS and UV acted as a pro-oxidant.M may be the premier molecule to protect the cells from oxidative stress.

Antioxidant capacity in postmortem brain tissues of Parkinson’s and Alzheimer’s diseases

Oxidative stress has been associated with damage and progressive cell death that occurs in neurodegenerative disorders such as Parkinsons disease (PD) and Alzheimers disease (AD). The aim of this study was to investigate the antioxidant capacity in postmortem motor cortex (MC), nucleus caudatus (NC), gyrus temporalis (GT) and substantia nigra (SN) from controls (C) and patients with PD and AD. The initial samples consisted of 68 subjects of PD, AD and C. Brains were matched for age, sex and postmortem time. Brain tissue was homogenized in a phosphate buffer pH 7.3 and separated with two-step centrifugation at 15,000rpm for 30 min and 15,000 rpm for 10 min at 4 degrees C. Antioxidant capacity in the supernatants was measured using the oxygen radical absorbance assay (ORAC). The results showed that in the SN of parkinsonians brain the balance between production of free radicals and the neutralization by a complex antioxidant system is disturbed. No changes in the antioxidant capacity of postmortem MC and NC of parkinsonians brain in comparison with C were found. In the SN of parkinsonians brain, antioxidant capacity seems to be lower in comparison with C (p < 0.05). Antioxidant capacity against peroxyl radical showed that MC of AD patients was lower than in the MC of C (p < 0.005). In NC of AD patients the antioxidant capacity against hydroxyl radical was increased in comparison with C (p < 0.04). No changes in the antioxidant capacity were found in brain tissues of AD in comparison with C, when CuSO4 was used as a free radical generator.

Brain antioxidant capacity in rat models of betacytotoxic-induced experimental sporadic Alzheimer’s disease and diabetes mellitus

It is believed that oxidative stress plays a central role in the pathogenesis of metabolic diseases like diabetes mellitus (DM) and its complications (like peripheral neuropathy) as well as in neurodegenerative disorders like sporadic Alzheimers disease (sAD). Representative experimental models of these diseases are streptozotocin (STZ)-induced diabetic rats and STZ-intracerebroventricularly (STZ-icv) treated rats, in which antioxidant capacity against peroxyl (ORAC(-ROO)*) and hydroxyl (ORAC(-OH)*) free radical was measured in three different brain regions (hippocampus, cerebellum, and brain stem) by means of oxygen radical absorbance capacity (ORAC) assay. In the brain of both STZ-induced diabetic and STZ-icv treated rats decreased antioxidant capacity has been found demonstrating regionally specific distribution. In the diabetic rats these abnormalities were not associated with the development of peripheral diabetic neuropathy. Also, these abnormalities were not prevented by the icv pretreatment of glucose transport inhibitor 5-thio-D-glucose in the STZ-icv treated rats, suggesting different mechanism for STZ-induced central effects from those at the periphery. Similarities in the oxidative stress alterations in the brain of STZ-icv rats and humans with sAD could be useful in the search for new drugs in the treatment of sAD that have antioxidant activity.

Antioxidant capacity in rat brain after ICV treatment with streptozotocin and alloxan — a preliminary study

Intracerebroventricular (icv) administration of betacytotoxic drug streptozotocin (STZ) produces long-term and progressive cognitive deficits in rats, as well as deficits in cerebral glucose and energy metabolism. These changes resemble those found in the brain of patients with sporadic Alzheimer’s disease (sAD), and therefore, STZ-icv treated rats have been proposed as an experimental model of sAD. In this study the antioxidant capacity (AC), using manual oxygen radical absorbance capacity (ORAC) assay, was measured in the rat brain frontoparietal cortex (FC) and brainstem-cerebellum region (BS-CB) after administration of STZ and another betacytotoxic drug alloxan (AL). Region-specific differences of AC were found, which were more expressed when hydroxyl radical (ORAC-OH°) generator was used in the assay. AC against ORAC-OH° was significantly lower in BS-CB than in FC of the control rats. Furthermore, ORAC-OH° significantly decreased in BS-CB 3-months following the icv administration of AL, but significantly increased following the TG+AL combined treatment in comparison with the controls. However, 3-months following the icv treatment of AL combination with a different glucose transport inhbitor, 3-O-methyl-D-glucose, ORAC-OH° values in BS-CB and ORAC-ROO° values in FC were significantly decreased in comparison to the controls. Our results suggest that betacytotoxic-icv treatment alters antioxidant defense systems in the brain, which particularly regarding the STZ-icv treatment, could be a useful tool in search for possible new antioxidant treatments of the neurodegenerative disorders such as sAD.

The Relation of Erythropoietin Towards Hemoglobin and Hematocrit in Varying Degrees of Renal Insufficiency.

Introduction: Hypoxia is a basic stimulant in production of erythropoietin (EPO). The primary function of erythrocytes is the transport of oxygen to tissues. Erythropoietin stimulates erythropoiesis which leads to increased production of erythrocytes- their total mass. This increases the capacity of the blood to carry oxygen, reduces the hypoxic stimulus and provides a negative feedback of stopping EPO production. The aim of this study was to establish a quantitative relationship between the concentration of erythropoietin, hemoglobin and hematocrit in different values of renal insufficiency. Material and methods: The survey was conducted on 562 subjects divided into two groups: with and without renal insufficiency. EPO, hemoglobin, hematocrit, serum creatinine and additional parameters iron, vitamin B12, and folic acid were determined by using immunochemical and spectrophotometric methods and glomerular filtration rate (GFR) was calculated as well. Results: EPO values (median) grow to the first degree of renal insufficiency, as compared to EPO values of healthy subjects, this increase is statistically significant, p=0.002. With further deterioration of renal function the values of EPO between all pathological groups are decreasing, and this decrease is statistically significant between first and second degree of renal insufficiency (RI) p<0.001. In the group of healthy subjects EPO is correlated rho = -0.532, p <0.0005 with hematocrit. The correlations are negative and strong and can be predicted by regression line (EP0 = 41.375- Hct * .649; EPO = 61.41–Hb * 0.355). In the group of subjects with the first degree of renal insufficiency EPO is in correlation with hematocrit rho=-0.574, p<0, 0005. It is also correlated with hemoglobin rho=-0.580, p< 0.0005. The correlation is negative (EP0= 42.168- Hct * 0.678). In the group of subjects with the third degree of renal insufficiency EPO is in correlation with hemoglobin rho=0.257, p=0.028. The correlation is medium strong and positive. In the group of subjects with third and fourth degree of renal insufficiency EPO is not in correlation with hemoglobin and hematocrit p>0.05. Conclusion: Renal dysfunction, depending on the level of RI effects differently on the biosynthesis of EPO in a diseased kidney, and consequently it also has a different effect on biosynthesis of HB in bone marrow and its content in the blood.

Interpretation of Erythropoietin and Haemoglobin Levels in Patients with Various Stages of Chronic Kidney Disease

Summary Background: The production of erythrocytes is regulated by the hormone erythropoietin (EPO), which maintains the blood haemoglobin (Hb) levels constant under normal conditions. Human EPO is a glycoprotein hormone and its synthesis is controlled by the hypoxia-inducible transcription factor. The aim of this study was to establish EPO and Hb levels in patients with chronic kidney disease (CKD), as well as in control subjects, and to investigate the relationship between these parameters. Methods: This cross-sectional, observational study included 356 subjects with CKD divided into 4 subgroups according to their glomerular filtration rate (GFR). The control group consisted of 206 age and sex matched healthy subjects with GFR rate ≥90 mL/min/1.73 m2. EPO, Hb and serum creatinine levels were determined by using immunochemical and spectrophotometric methods. GFR was determined using the MDRD formula. Results: The CKD patients had significantly lower levels of haemoglobin (p<0.0005) and hematocrit (p<0.0005) compared to control group. Our results showed that Hb levels decreased, whereas serum creatinine increased with the increasing renal failure. The CKD patients in all four groups had significantly lower (p<0.0005) Hb levels, and significantly higher (p<0.0005) creatinine levels compared to the control group. The median EPO in group I and II were significantly higher (p=0.002; p=0.018), while median EPO in group III and IV were significantly lower (p=0.03; p=0.011) compared to the control group. Conclusions: In patients with CKD, GFR positively correlated with Hb and EPO, while the correlation between GFR and serum creatinine was negative.

Hydrophilic antioxidant scores against hydroxyl and peroxyl radicalsin honey samples from Bosnia and Herzegovina

The aim of this study was to evaluate the total hydrophilic antioxidant scores (THAS) against peroxyl (ROO•) and hydroxyl (OH•) free radicals in different types (d.t.) of bee honey from Bosnia and Herzegovina (B&H). It has been analyzed 46 honey samples of d.t. such as: meadow honey (MeH), acacia honey (AH), forest honey (FH), mountain honey (MoH), heather honey (HH), and chestnut honey (CH). All analyses were performed by oxygen radical absorbance capacity (ORAC) assay, using trolox as a standard. Antioxidant capacity (AC) against ROO• and OH• (AC ROO • and AC OH • ) expressed as trolox equivalents per honey weight (mM TE/g): as the THAS [(AC (LA+HA)ROO • +AC (LA+HA)OH • ), AC derived from low-molecular and high-molecular weight antioxidants (LA and HA, respectively) together in the bulk], then as the HAS against ROO• and OH• derived from LA only (HAS(LA)), and as the HAS against ROO• and OH• derived from HA only (HAS(HA)). ORAC analysis showed that THAS d.t. of honeys decreased as follows FH>HH>AH>MoH>MeH>CH, then HAS(LA) d.t. of honeys decreased as follows MoH>FH>AH>HH>MeH>CH. Also, HAS(HA) for AH was higher than HAS(HA) for MeH.

Correlation of Total Secondary Sulfur Compounds, Total Phenols and Antioxidant Capacity in the Ramsons and Garlic

Aims: This study explores the total quantity of sulfur secondary metabolites in the plant organs of garlic and ramsons, the content of total phenol and their correlation to the antioxidant capacity (AOC). There are different reports about correlation of secondary