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.

Evaluation of phenolic profile, enzyme inhibitory and antimicrobial activities of Nigella sativa L. seed extracts.

Black cumin (Nigella sativa L. [N.sativa]) seed extracts demonstrated numerous beneficial biological effects including, among others, antidiabetic, anticancer, immunomodulatory, antimicrobial, anti-inflammatory, antihypertensive, and antioxidant activity. To better understand the phytochemical composition of N. sativa seeds, methanol seed extracts were analyzed for phenolic acid and flavonoid content. Furthermore, we tested N. sativa methanol, n-hexane, and aqueous seed extracts for their inhibitory activity against butyrylcholinesterase (BChE) and catalase (CAT) as well as for antimicrobial activity against several bacterial and a yeast strains. The phenolic content of N. sativa was analyzed using ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). The inhibition of BChE was assessed by modified Ellman's method, and the inhibition of CAT was determined by monitoring hydrogen peroxide consumption. The extracts were tested against Bacillus subtilis, Staphylococcus aureus, Salmonella enterica, and Escherichia coli using the agar diffusion method. The UHPLC-MS/MS method allowed the identification and quantification of 23 phenolic compounds within 15 minutes. The major components found in N. sativa seed extract were sinapinic acid (7.22 ± 0.73 µg/mg) as a phenolic acid and kaempferol (11.74 ± 0.92 µg/mg) as a flavonoid. All extracts showed inhibitory activity against BChE, with methanol seed extract demonstrating the highest inhibitory activity (inhibitory concentration 50% [IC50] 79.11 ± 6.06 µg/ml). The methanol seed extract also showed strong inhibitory activity against CAT with an IC50 value of 6.61 ± 0.27 µg/ml. Finally, the methanol extract exhibited considerable inhibitory activity against the tested microbial strains. Overall, this is the first study to investigate the ability of black cumin seed extracts to inhibit CAT. Our results indicate that N. sativa seed can be considered as an effective inhibitor of CAT activity.

Chemical composition of various Ephedra species.

The medicinal significance of Ephedra is based on the sympathomimetic properties of ephedrine (E) alkaloids. Pharmacological effects depend on the phytocomposition of individual Ephedra species. The aim of this study was to measure the total alkaloids content (TAC), total phenolics content (TPC), and total flavonoids content (TFC) and determine their relationship in dry herb of Ephedra major, Ephedra distachya subsp. helvetica, Ephedra monosperma, Ephedra fragilis, Ephedra foeminea, Ephedra alata, Ephedra altissima and Ephedra foliata. Nowadays, medicinal use of Ephedrae herba is limited, but the abuse of its psychostimulants is rising. In this study, TAC, TPC and TFC were determined using spectrophotometric methods. For the first time, ultra-performance liquid chromatography with ultraviolet detection (UPLC-UV) was used for separation and quantification of E-type alkaloids of various Ephedra species. The highest TPC and TFC were found in E. alata (53.3 ± 0.1 mg Gallic acid equivalents/g dry weight, 2.8 mg quercetin equivalents/g dry weight, respectively). The total content of E and pseudoephedrine determined by UPLC-UV varied between 20.8 mg/g dry weight (E. distachya subsp. helvetica) and 34.7 mg/g dry weight (E. monosperma). The variable content and ratio between secondary metabolites determined in different Ephedra species reflects their metabolic activities. Utilization of UPLC-UV unveiled that this technique is sensitive, selective, and useful for separation and quantification of different alkaloids in complex biological matrixes. The limit of detection was 5 ng. Application of UPLC-UV can be recommended in quick analyses of E-type alkaloids in forensic medicine and quality control of pharmaceutical preparations.

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.

Anticoagulant activity of some Artemisia dracunculus leaf extracts.

Platelet hyperactivity and platelet interaction with endothelial cells contribute to the development and progression of many cardiovascular diseases such as atherosclerosis and thrombosis. The impact of platelet activity with different pharmacological agents, such as acetylsalicylic acid and coumarin derivatives, has been shown to be effective in the prevention of cardiovascular disease. Artemisia dracunculus, L. Asteraceae (Tarragon) is used for centuries in the daily diet in many Middle Eastern countries, and it is well known for its anticoagulant activity. The present study investigates the presence of coumarins in tarragon leaves and subsequently determines the extract with a major amount of coumarin derivatives. The solvents of different polarities and different pH values were used for the purpose of purifying the primary extract in order to obtain fractions with the highest coumarin content. Those extracts and fractions were investigated for their anticoagulant activity by determining prothrombin time (PT) and the international normalized ratio (INR), expressed in relation to the coagulation time of the healthy person. Purified extracts and fractions obtained from plant residue after essential oil distillation, concentrated in coumarin derivatives, showed the best anticoagulant activity, using samples of human blood. INR maximum value (2.34) and consequently the best anticoagulant activity showed the methanol extract at concentration of 5%.  The INR value of normal plasma in testing this extract was 1.05.

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.

Elevated serum homocysteine level is not associated with serum C-reactive protein in patients with probable Alzheimer's disease.

Elevated plasma homocysteine (Hcy) levels have been associated with Alzheimer's disease (AD) and cognitive impairment. Studies have shown that Hcy may have direct and indirect neurotoxicity effects. The aim of the study was to investigate serum Hcy concentration in patients with probable AD with age-matched controls and to determine whether there was an association between serum Hcy and C-reactive protein concentration in patients with probable AD. We also aimed to determine whether there was an association between serum tHcy concentration and cognitive impairment in patients with probable AD. Serum concentration of total Hcy was determined by the fluorescence polarization immunoassay on the AxSYM system, and serum C-reactive protein (CRP) concentration was determined by means of particle-enhanced immunonephelometry with the use of BN II analyzer. Cognitive impairment was tested by the MMSE score. Body mass index (BMI) was calculated for each subject included in the study. Age, systolic and diastolic blood pressure and BMI did not differ significantly between the two groups. Mean serum tHcy concentration in the control group of subjects was 12.60 mumol/L, while in patients with probable AD the mean serum tHcy concentration was significantly higher than 16.15 mumol/L (p < 0.01). A significant negative association between serum tHcy concentration and cognitive impairment tested by the MMSE score in patients with probable AD was determined (r = -0.61634; p < 0.001). Positive, although not significant correlation between CRP and serum tHcy concentrations in patients with AD, was observed. Increased tHcy concentration in patients with probable AD, and the established negative correlation between serum tHcy concentration and cognitive damage tested by MMSE score in the same group of patients, suggests the possible independent role of Hcy in the pathogenesis of AD and cognitive impairment associated with this disease.

Identification and isolation of pharmacologically active triterpenes in Betuale cortex, Betula pendula Roth., Betulaceae.

Betulae cortex, Betula pendula Roth., Betulaceae, comprise triterpene substances which are confirmed to posses very important pharmacological activities such as anti-inflammatory, anticancer and antiviral. In this study, extraction of triterpene substances from both, inner and external birch bark was carried out and after that qualitative analysis on betulin, betulinic acid, oleanolic acid and lupeol was performed by method of thin layer chromatography. By this separation method, applying system for development benzene-ethyl acetate-formic acid (36:12:5), is gained a good separation of examined triterpene substances from methanol extracts of inner and external birch bark as well as used standards. From obtained row triterpene mixtures, certain triterpene substances are isolated using method of dry column chromatography. To those substances infrared (IR) spectra were recorded and compared with IR spectra of adequate standards. The study encloses all obtained IR spectra and interpretations on the basis of which can be concluded that triterpene substances, betulin, betulin acid and lupeol isolated from external birch bark give identical characteristic signals and absorbance as referent standards. Method of dry column chromatography has resulted as simple, efficient, repeatable and economical for laboratory conditions. Beside this, a sufficient quantity of examined triterpene substances is also obtained for continuation of their further analytical analysis.

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.

Serum prolactin, leptin, lipids and lipoproteins levels during antipsychotics treatment in Parkinson's disease and related psychosis.

Weight gain is a common adverse effect associated with the use of most typical and atypical antipsychotic. Aim of this study was to investigate serum prolactin, leptin, cholesterol, triglyceride, lipoproteins, such high density lipoprotein (HDL), and low density lipoprotein (LDL) levels in patients with Parkinson's disease (PD)-related psychosis during long-term medication with atypical antipsychotic. The study population comprised 40 patients, who were divided into 4 groups: olanzapine (n=10), risperidone (n=10), seroquel (n=10) monotherapy, a group of 10 patients receiving only antiparkinson drugs and a control group of 8 healthy persons. The patients were evaluated at baseline and at the sixth and twelfth week according to the Positive and Negative Syndrome Scale (PANSS), body mass index (BMI), and fasting serum prolactin, leptin, lipids and lipoproteins levels. Treatment of patients with olanzapine caused marked increase of serum LDL, cholesterol, triglyceride, and leptin levels (p<0,02). No changes in HDL concentrations. There was positive relationship between serum leptin, lipid levels and BMI. However, treatment of patients with seroquel did not cause changes in serum prolactin, leptin, lipids, and lipoproteins levels. Our results suggest that treatment of patients with PD-related psychosis with seroquel appears to have minimal influence on serum leptin, prolactin, lipids, lipoproteins and BMI compared with olanzapine and risperidone.

Amperometric determination of bonded glucose with an MnO(2) and glucose oxidase bulk-modified screen-printed electrode using flow-injection analysis.

A screen-printed amperometric biosensor based on carbon ink double bulk-modified with MnO(2) as a mediator and glucose oxidase as a biocomponent was investigated for its ability to serve as a detector for bonded glucose in different compounds, such as cellobiose, saccharose, (-)-4-nitrophenyl-beta-d-glucopyranoside, as well as in beer samples by flow-injection analysis (FIA). The biosensor could be operated under physiological conditions (0.1M phosphate buffer, pH 7.5) and exhibited good reproducibility and stability. Bonded glucose was released with glucosidase in solution, and the free glucose was detected with the modified screen-printed electrode (SPE). The release of glucose by the aid of glucosidase from cellobiose, saccharose and (-)-4-nitrophenyl-beta-d-glucopyranoside in solution showed that stoichiometric quantities of free glucose could be monitored in all three cases. The linear range of the amperometric response of the biosensor in the FIA-mode flow rate 0.2mLmin(-1), injection volume 0.25mL, operation potential 0.48V versus Ag/AgCl) extends from 11 to 13,900mumolL(-1) glucose in free form. The limit of detection (3sigma) is 1mumolL(-1) glucose. A concentration of 100mumolL(-1) yields a relative standard deviation of approximately 7% with five injections. These values correspond to the same concentrations of bonded glucose supposed that it is liberated quantitatively (incubation for 2h with glucosidase). Bonded glucose could be determined in beer samples using the same assay. The results corresponded very well with the reference procedure.

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.

A placebo-controlled study of memantine (Ebixa) in dementia of Wernicke-Korsakoff syndrome.

We evaluated the responses of 16 patients to preliminarily explore the spectrum of effectiveness and tolerability of the memantine, and NMDA antagonist, in the treatment of dementia in Wernicke-Korsakoff syndrome. In this study, for the first time in dementia of Wernicke-Korsakoff syndrome, the response to memantine was assessed. 16 patients with median age of 64 years and median body weight of 77 kg were treated with memantine 10 mg twice daily for up to 28 weeks. Clinical global impressions (CGI), and Mini Mental Status Examination (MMSE) were performed during the treatment period (after 2, 4, and 28 weeks). Efficacy measures also included the ADCS-Activities of Daily Living scale (ADCS-ADL). At 28 weeks, the ADCS-ADL showed significantly less deterioration in memantine treated patients compared with placebo (-2.3 compared with -4.3: p = 0.005). The results of MMSE demonstrate a significant and clinically relevant benefit for memantine relative to placebo as shown by positive outcomes in cognitive and functional assessments. Memantine (10 mg) was safe and well tolerated. The preliminarily findings of this study with 16 patients suggested that memantine is effective in the treatment of dementia in Wernicke-Korsakoff syndrome.

A pilot study of Topiramate (Topamax) in the treatment of tonic-clonic seizures of alcohol withdrawal syndromes.

In this study for the first time in alcohol withdrawal syndrome, the response to topiramate was assessed. 12 patients with median age of 49.5 years and median body weight of 76.3 kg were treated with topiramate twice daily for up 30 days, starting with a dose of 50 mg in the morning and 50 mg in the evening. The preliminary findings of this study suggest that topiramate is very effective against tonic-clonic seizures in alcohol withdrawal syndrome. No side effects were observed. Only two patients had loss of body weight (3-3.5 kg/4 weeks).