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

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.

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.

Serum nitric oxide concentrations in patients with multiple sclerosis and patients with epilepsy.

Nitric oxide (NO), a neurotransmitter and a free radical, has been purported to be involved in numerous neurological diseases. We investigated the serum nitric oxide concentration in 30 patients with multiple sclerosis (MS), in 30 patients with epilepsy and in 30 control subjects. The aim was also to determine whether a statistically significant difference in serum NO concentrations exists between the groups of interest. The total serum nitric oxide concentration was measured using the Griess reaction after reducing nitrates to nitrites with elemental zinc. In the group multiple sclerosis, the mean NO concentrations were X ± SEM = 31.02 ± 1.79 µmol/l, in the control group X ± SEM = 25.31 ± 1.44 µmol/l and in the group epilepsy X ± SEM = 22.51 ± 1.28 µmol/l. Student's t test showed a statistically significant difference between subjects with multiple sclerosis and the control group (p = 0.013), as well as between the groups multiple sclerosis and epilepsy (p = 0.0002). This data confirms that NO may play an important role in the pathogenesis of multiple sclerosis, whereas its role in epilepsy still remains unclear.

Daily fluctuation of cortisol in the saliva and serum of healthy persons.

UNLABELLED: The objective of this study was to evaluate cortisol in the saliva and serum of healthy persons and its daily fluctuations by using immunochemical method on a autoanalyzer. Biological samples: Serum from 14 healthy persons and saliva from 18 healthy persons were taken two times at 8 a.m. and at 4 p.m. Immunochemical assay: The principle of this method is the competitive binding of cortisol present in the analyzed sample and cortisol marked with peroxides on binding parts with specific antibodies. STATISTICAL ANALYSIS: Student t-test. Cortisol in saliva in the morning: 21,2 +/- 16,2 nmol/l and in the afternoon 12,7 +/- 8,1 nmol/l. Cortisol in serum in the morning: 459, 6 +/- 235,2 nmol/l , and in the afternoon 340,5 +/- 207,5 nmol/l. The concentrations of cortisol in saliva are lower than in serum. Cortisol in the serum in the morning is about twenty times higher than cortisol in the saliva at the same time. Cortisol in the serum at afternoon is about twenty-seven times higher than cortisol in the saliva. Individual variabilities of cortisol in the saliva and serum were found during the day.

Urinary hippuric acid after ingestion of edible fruits.

Aim of this study was to evaluate the biotransformation of simple phenols after ingestion of edible fruits and mixed food. It was analyzed hippuric acid in urine as biomarker of conjugation in the liver cells of glycine with aromatic phenolic acids such benzoic and salicylic acid from ingested food. Measurement of hippuric acid in urine samples of 10 healthy individuals: 5 female and 5 male with a mean age 51,5 years were recruited to participate in this study. Urine samples were collected for 24 hours. The additional meals 300 g of fruits: blueberry, cherry, raspberry, melon, blackberry and mixed food were given immediately before the 24 hr urine sampling. Otherwise, the meals given during 24 hr was a usually food. Biotransformation of phenols in edible fruits, that are together with liver glycins precursors of hippuric acid biosynthesis, was evaluated by direct spectrophotometric measurement of excreted hippuric acid in urine at 410 nm. It was established that the highest quantity of hippuric acid was after ingestion of 300 g of bilberry fruits (p< 0,003), and same quantity of cherries (p< 0,003). Concentration of excreted hippuric acid was twice higher after ingestion of these fruits in comparison with hippuric acid concentrations in urine after ingestion of common - mixed food. Quantity of biosynthesised hippuric acid was in direct correlation with the concentrations of its precursors, primarily phenol acids and other simple aromatic acids ingested with food.

Antioxidant capacity in the lipophilic fraction of Alzheimer's brain tissues.

The aim of this study was to investigate the antioxidant capacity (AC) in the lipophilic fraction of postmortem motorcortex (MC), nucleus caudatus (NC) and gyrus temporalis (GT) from controls (C) and Alzheimer's disease (AD) patients. The initial samples consisted of 50 human brain tissues of AD and C. AC of the different region of human brain were measured by using the fluorescent method of the oxygen radical absorbance capacity (ORAC). Peroxyl and hydroxyl radical generators were used in the analysis. All ORAC analysis were carried out on the Perkin-Elmer spectrofluorometer LS 55 with fluorescent filters, Ex: 485 nm; Em: 520 nm. Final results were calculated using the differences between area under the quenching curve of fluorescein (FL), blank and analyzed biological samples. AC against peroxyl radicals (ORAC-ROO degrees ) of lipophilic fraction in MC of AD was statistically significantly lower in comparison with MC of C (p < 0.008). No changes in the AC against hydroxyl radicals (ORAC- degrees OH) of lipophilic fraction of AD were found in comparison with C. Reduction of total protein in GT of AD (p < 0.03) was found. The results showed that in the MC of AD brain the balance between production of free radicals and the neutralization by a complex antioxidant system is disturbed. The manual fluorescent method for AC measurements proved to be sufficiently appropriate and sensitive for the AC measurements of lipophilic fraction of postmortem brain tissues from different patologic conditions.

Plasma lipids, lipoproteins and hormones levels during olanzapine treatment in psychosis and depression.

Weight gain is common adverse effect associated with the use of most typical and atypical antipsychotic. Aim of this study was to investigate plasma lipids, lipoproteins and some hormones levels during olanzapine treatment in patients with psychosis. The study population comprised twenty nine patients (29) diagnosed with psychosis and eleven patients (11) with endogenous depression. Plasma cholesterol, triglicerides, phospholipids, high-density lipoprotein cholesterol (HDL-C), very-low-density lipoprotein cholesterol (VLDL-C), low-density lipoprotein cholesterol (LDL-C), apolipoprotein A-I (ApoA-I) and apolipoprotein B (Apo B), and hormones - prolactin, cortisol, follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone and estradiol were measured by auto-analyzer and by classic photometric methods. All measurements were performed before and during one-year treatment with olanzapine and antidepressant. Treatment of the schizophrenic patients with olanzapine caused a great metabolic impact that is primarily expressed in body mass, cholesterol increase and statistically significant rise of BMI, respectively. Body mass increase could be explained by the fact that olanzapine blocked not only dopaminergic, serotonergic, cholinergic, alpha-adrenergic but histaminergic receptors as well.

Reduced brain antioxidant capacity in rat models of betacytotoxic-induced experimental sporadic Alzheimer's disease and diabetes mellitus.

It is believed that oxidative stress (OS) 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 Alzheimer's 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 (AC) against peroxyl (ORAC(-ROO) (*)) and hydroxyl (ORAC(-OH) (*)) free radicals (FR) was measured in three different brain regions: the hippocampus (HPC), the cerebellum (CB), and the brain stem (BS) by means of oxygen radical absorbance capacity (ORAC) assay. In the brain of both STZ-induced diabetic and STZ-icv treated rats decreased AC has been found demonstrating regionally specific distribution. In the diabetic rats these abnormalities were not associated with the development of peripheral diabetic neuropathy (PDN). Also, these abnormalities were not prevented by the intracerebroventricularly (icv) pretreatment of glucose transport inhibitor 5-thio-D: -glucose (TG) in the STZ-icv treated rats, suggesting different mechanism of STZ-induced central effects from those at the periphery. Similarities of the OS alterations in the brain of STZ-icv rats and humans with sAD could be useful in the search for the new drugs in the treatment of sAD that have antioxidant activity. In the STZ-induced diabetic animals the existence of PDN was tested by the paw pressure test, 3 weeks following the diabetes induction. Mechanical nociceptive thresholds were measured three times at 10-min intervals by applying increased pressure to the hind paw until the paw-withdrawal or overt struggling was elicited. Only those diabetic animals which demonstrated decreased withdrawal threshold values in comparison with the control non-diabetic animals (C) were considered to have developed the PDN.

Total content of phenols and anthocyanins in edible fruits from Bosnia.

Content of total phenols and total anthocyanins was estimated in edible fruits from Bosnia by photometric methods. Cyanidin-3-galactoside chloride was used as a standard for determination of total anthocyanins, and galic acid served as a standard for determination of total phenols. Total content of phenols was 12.7 mg/g in elderberry fruits, 10.4 mg/g in bilberry, 9.8 mg/g in blackberry, 8.8 mg/g in wild cherry, 6.1 mg/g in cultivated blackberry, 3.5 mg/g in cultivated strawberry, 2.4 mg/g in average in sour cherry fruits from different locations and the lowest quantity of total phenols was in edible parts of melon, only 0.2 mg/g. Total content of anthocyanins was 6.8 mg/g in wild cherry, 6.7 mg/g in elderberry fruits and 4.5 mg/g in bilberry. Wild bilberry fruits from different locations had in average 3.5 mg/g, cherries from different locations 1.3 mg/g, cultivated blackberries 1.0 mg/g, cultivated strawberries 0.8 mg/g while melon fruit had no anthocyanins at all. Acidity was measured in macerate of edible fruits by direct insertion of electrode. pH values in the macerates were as follows: 3.03 in bilberry, 3.45 in blackberries, 3.59 in sour cherries, 3.92 in wild cherries, 4.44 in elderberries and 6.19 in melon.

Inhibition of Neutral red photolysis with different antioxidants.

Neutral red is a dye the azine structure which has been used as an acido-base indicator and a dye in histochemistry. In 1960 Goldhaber introduced Neutral red into the medium of resorbing bone cultures to localize the osteoclast in the living cultures. Using time-lapse microcinematography in order to follow the osteoclasts, he reported excellent contrast could be obtained with Neutral red due to the avidity of osteoclasts for this dye. Unfortunately, however, the photodynamic effect resulting from subsequent exposure of these cultures to light precluded this approach, and again in 1963. it was observed that the death of the osteoclasts was probably due to a photodynamic effect related to the dye in the cell, the presence of oxygen and the frequent exposure of light by our time-lapse photography. VIS and UV irradiation induced photolysis of Neutral red, and from Neutral red cation produced with photons a Neutral red radical. This Neutral red radical can be inhibited with action of an antioxidant, such as melatonin, glutathione, ascorbic acid, E vitamin, etc. We developed an assay with Neutral red photolysis which utilizes a VIS and UV irradiation technique for quantification the inhibition of photolysis with action of an antioxidant. In this method Neutral red acts double, as a free radical generator and as a photosensitizer.

A study of gabapentin in the treatment of tonic-clonic seizures of alcohol withdrawal syndrome.

In this study for thirty (30) patients with alcohol withdrawal syndrome, the response to anticolvusant gabapentin was assessed. Thirty (30) patients with median age of 57.0 years and median body weight of 79.1 kg were treated with gabapentin 3 x 300 mg daily for up 30 days. The preliminary findings of this study suggest that gabapentin is very effective against tonic-clonic seizures in alcohol withdrawal syndrome. Gabapentin was safe and well tolerated. For twenty (20) patients no side effect were observed.