Low-density lipoprotein cholesterol-to-apolipoprotein B ratio as a potential indicator of LDL particle size and plasma atherogenicity in type 2 diabetes.

AIMS: Atherogenic dyslipidemia, associated with small, dense low-density lipoprotein-cholesterol (S-LDL) particles and impaired metabolism of triglycerides (TGs) and high-density lipoprotein-cholesterol (HDL-c), leads to the development of atherosclerosis-related complications of type 2 diabetes mellitus. Based on the hypothesis that an LDL-c-to-apolipoprotein B ratio (LDL/ApoB) < 1.2 may predict the prevalence of S-LDL, this study aimed to evaluate the LDL/ApoB ratio in patients with type 2 diabetes with moderately elevated TG levels. METHODS: The study population consisted of 121 outpatients with type 2 diabetes (S-LDL group, LDL/ApoB < 1.2, n = 79; L-LDL group, LDL/ApoB > 1.2, n = 42) and 58 healthy subjects. The LDL/ApoB ratio was calculated from the measured LDL-c and ApoB levels in participants with TG levels lower than 4.5 mmol/L. Since TGs and HDL-c are included in the atherogenic index of plasma (AIP), we evaluated the relationship between LDL/ApoB and the AIP. RESULTS: Higher levels of AIP, TG (both P < 0.0001), and lipid hydroperoxides (LOOH) (P < 0.001) and lower levels of HDL-c, total cholesterol, and non-HDL-c (P < 0.001, <0.01, <0.05, respectively) were found in the S-LDL group compared to the L-LDL group. There were significant relationships between the LDL/ApoB ratio and the AIP, TG (both P < 0.0001), LOOH (P < 0.0005), and HDL-c levels (P < 0.05) in the S-LDL group. CONCLUSIONS: The prevalence of S-LDL particles (65%) and the close association of LDL/ApoB with the AIP suggest that this ratio may be a potential indicator of increased cardiovascular risk in patients with type 2 diabetes.

Abnormalities in the relationship of paraoxonase 1 with HDL and apolipoprotein A1 and their possible connection to HDL dysfunctionality in type 2 diabetes.

AIMS: Lipid parameters, lipid risk indexes and lipid-related oxidative stress markers (paraoxonase 1 [PON1] and lipid peroxides [LPO]) reflect the actual status of lipid metabolism in type 2 diabetes (T2DM). We hypothesized that relationships of high-density lipoprotein cholesterol (HDL-c) with PON1 and apolipoprotein A1 (ApoA1) and/or PON1 with ApoA1 under conditions of hyperglycaemia and oxidative stress might reveal HDL functionality. We investigated relationships between PON1, LPO, and lipid risk markers in T2DM subjects and compared them with those in healthy subjects. METHODS: A total of 107 Caucasian subjects, 67 T2DM outpatients (mean age 52.2 ±â€¯6.9 years) and 40 healthy subjects (mean age 48.1 ±â€¯7.5 years) were included in the study. Serum levels of total cholesterol (CHOL-T), HDL-c, low-density lipoprotein cholesterol (LDL-c), triglycerides (TG), apolipoprotein B (ApoB), ApoA1, LPO, and PON1 activity were measured. Non-HDL-c, ApoB/ApoA1 and non-HDL/HDL (lipid risk indexes) were calculated. RESULTS: Higher levels of TG, LPO (P < 0.0001), nonHDL/HDL and ApoB/ApoA1 (P < 0.001, 0.05, respectively), and lower levels of HDL-c, ApoA1, and PON1 (P < 0.0001) were observed in T2DM subjects than in controls. There is a lack of relationship among PON1, HDL-c, and ApoA1 in T2DM patients. PON1 activity positively correlated with these parameters (P < 0.0001) in controls. Strong correlations between non-HDL-c and ApoB (r = 0.956 vs. 0.756; P < 0.0001), LPO and TG (r = 0.831 vs. 0.739; P < 0.0001) were recorded in both study groups (P < 0.0001). CONCLUSIONS: Impaired anti-oxidant and anti-atherogenic HDL properties associated with weakened PON1 function and lipid peroxidation may contribute to the development of atherosclerosis-related diseases in T2DM.

Relationship between selected oxidative stress markers and lipid risk factors for cardiovascular disease in middle-aged adults and its possible clinical relevance.

OBJECTIVES: Decreased activity of HDL-associated paraoxonase 1 (PON1) and elevated levels of lipid peroxides together with abnormal lipid profile may prognosticate the progression of atherosclerosis. The aim of this study was to assess associations between selected oxidative stress markers (PON1, lipid peroxides) and lipid risk factors for cardiovascular disease in middle-aged subjects. DESIGN AND METHODS: Arylesterase activity of PON1; lipid peroxides; total-, HDL-, LDL-, non-HDL-cholesterol; triglycerides; apolipoproteins A-I (ApoA-I) and B (ApoB), and lipid risk indexes were determined in serum of 75 volunteers (mean age 41.7±8.2years). Forty six volunteers were divided into normolipidemic (NL) and hyperlipidemic (HL) group. RESULTS: Elevated levels of atherogenic cholesterol (LDL, non-HDL), lipid risk indexes (p<0.0001), lipid peroxides (p<0.001), and decreased activity of PON1 (p<0.05) were found in HL group. Strong correlations between PON1 activity and HDL (r=0.635, p=0.0005), apolipoprotein A-I (r=0.703, p<0.0001), ApoA-I/ApoB ratio (r=0.536, p=0.004), and non-HDL/HDL ratio (r=-0.445, p=0.013) were observed in NL group. There was no significant association between PON1 activity and these parameters in HL group. CONCLUSIONS: Significant abnormalities of lipid parameters, oxidative stress markers and associations between PON1, HDL and apolipoprotein A-I may influence the antioxidant and anti-atherogenic functions of HDL and result in the higher susceptibility of lipoproteins to oxidative modification. Monitoring of lipid profile together with oxidative stress markers in an asymptomatic population could be beneficial for early identification of atherosclerosis-related diseases.

[Significance of alanine aminotransferase screening in blood donors for risk reduction of hepatitis B and C transmission by haemotherapy]. / Význam vysetrovania alanínaminotransferázy u darcov krvi pre redukciu rizika prenosu hepatitíd B a C hemoterapiou.

BACKGROUND: The goal of the retrospective observatory cross-sectional study was to evaluate the benefit of alanine aminotransferase screening of blood donors in prevention of hepatitis B and C transmission by haemotherapy in context of actual screening methods. METHODS: Donations with elevated ALT more than the defined limit (ALT men 80 IU/l, women 64 IU/l, spectrophotometric UV test, KUADRO(TM), BPC BioSed Srt, Castelnuovo di Porto Roma, Italy) and/or reactivity any of the hepatitis screening parameters HBsAg, anti-HBc, anti-HCV (chemiluminescence method, ARCHITECT i2000(TM), Illinois, USA) were evaluated. Donors were confirmatory retested. They were classified into groups with common biological properties according to their final virological status and statistically evaluated in programs Graph Pad Prism 6.05 and Microsoft Excel 2003. RESULTS: From a total of 61 214 donations elevated ALT was found in 420 (0.69 %), active HBV in 25 (0.04 %), active HCV infection in 5 (0.01 %) blood donors. Coincidental elevation of ALT and active HBV infection occured in 1 donor (0.002 %), as well as HCV (0.002 %). Levels of ALT were higher in the group with elevated ALT without active HBV or HCV infection than in groups with active HCV and HCV infection (p < 0.05). Occurence of blood donor in seronegative anti-HCV window was not observed. Elevated ALT was low specific (69.14 %) and senzitive (6.45 %) for active hepatitis. We did not prove positive correlation of ALT and S/CO (signal-to-cut-off) anti-HBc (Spearman r = -0,565, p < 0.0001), ALT and S/CO anti-HCV (Spearman r = -0.1046, p = 0.0022), in ALT and S/CO HBsAg the result was not statistically significant (Spearman r = -0.00968, p = 0.77). Positive but statistically insignificant correlation ALT and S/CO anti-HCV occured in the group of 5 blood donors with active HCV infection (Spearman r = 0.4, p = 0.51). Screening scheme for HCV infection testing anti-HCV + ALT was per one donation by € 0.18 more expensive than the scheme anti-HCV + HCV RNA due to amount of waisted donations with ALT elevation (825 TU, € 41 388.89). CONCLUSION: Elevation of ALT in blood donors was not pathognomonic for hepatitis B and C infection. Screening of HCV consisting of anti-HCV + HCV RNA (nucleic acid testing method, COBAS AmpliScreen HCV 2.0(TM), ROCHE Diagnostics, Hague Road, Indianapolis, USA) is more cost-effective than the scheme anti-HCV + ALT.

Arbutin and decrease of potentially toxic substances generated in human blood neutrophils.

Neutrophils, highly motile phagocytic cells, constitute the first line of host defense and simultaneously they are considered to be central cells of chronic inflammation. In combination with standard therapeutic procedures, natural substances are gaining interest as an option for enhancing the effectiveness of treatment of inflammatory diseases. We investigated the effect of arbutin and carvedilol and of their combination on 4ß-phorbol-12ß-myristate-13α-acetate- stimulated functions of human isolated neutrophils. Cells were preincubated with the drugs tested and subsequently stimulated. Superoxide (with or without blood platelets, in the rate close to physiological conditions [1:50]) and HOCl generation, elastase and myeloperoxidase release were determined spectrophotometrically and phospholipase D activation spectrofluorometrically. The combined effect of arbutin and carvedilol was found to be more effective than the effect of each compound alone. Our study provided evidence supporting the potential beneficial effect of arbutin alone or in combination with carvedilol in diminishing tissue damage by decreasing phospholipase D, myeloperoxidase and elastase activity and by attenuating the generation of superoxide and the subsequently derived reactive oxygen species. The presented data indicate the ability of arbutin to suppress the onset and progression of inflammation.

The natural stilbenoid piceatannol decreases activity and accelerates apoptosis of human neutrophils: involvement of protein kinase C.

Neutrophils are able to release cytotoxic substances and inflammatory mediators, which, along with their delayed apoptosis, have a potential to maintain permanent inflammation. Therefore, treatment of diseases associated with chronic inflammation should be focused on neutrophils; formation of reactive oxygen species and apoptosis of these cells represent two promising targets for pharmacological intervention. Piceatannol, a naturally occurring stilbenoid, has the ability to reduce the toxic action of neutrophils. This substance decreased the amount of oxidants produced by neutrophils both extra- and intracellularly. Radicals formed within neutrophils (fulfilling a regulatory role) were reduced to a lesser extent than extracellular oxidants, potentially dangerous for host tissues. Moreover, piceatannol did not affect the phosphorylation of p40(phox)-a component of NADPH oxidase, responsible for the assembly of functional oxidase in intracellular (granular) membranes. The stilbenoid tested elevated the percentage of early apoptotic neutrophils, inhibited the activity of protein kinase C (PKC)-the main regulatory enzyme in neutrophils, and reduced phosphorylation of PKC isoforms α , ß II, and δ on their catalytic region. The results indicated that piceatannol may be useful as a complementary medicine in states associated with persisting neutrophil activation and with oxidative damage of tissues.

Effect of stilbene derivative on superoxide generation and enzyme release from human neutrophils in vitro.

Neutrophils represent the body's primary line of defense against invading pathogens. They most rapidly reach the site of injury or infection, liberate antimicrobial proteins, proteases and produce reactive oxygen species. Prolonged or excessive liberation of these very effective and toxic substances could intensify the inflammatory process and enhance tissue damage in many diseases, such as allergies, infections and rheumatoid arthritis. Pterostilbene belongs to stilbenoids, structural analogues of resveratrol, which act as natural protective agents in defending the plant against viral and microbial attack. It possesses anticancerous, antidiabetic and anti-inflammatory properties.The study provides new information on the effect of pterostilbene [0.01-100 µmol/l] on superoxide generation in and myeloperoxidase (MPO) release from azurophil granules of isolated human neutrophils. PMA [1µmol/l], which activates NADPH-oxidase via protein kinase C, was used for stimulation of neutrophils Unstimulated cells showed neither superoxide generation nor myelopereoxidase release after preincubation with the drug studied. Pterostilbene dose dependently decreased superoxide generation in and MPO release from stimulated human neutrophils, however a significant decrease was recorded only in the concentration 100 µmol/l. The effect of pterostilbene was more pronounced on superoxide generation in comparison to MPO release. Our results suggest that the effect of pterostilbene may prove beneficial in controlling inflammation.

Quercetin inhibits degranulation and superoxide generation in PMA stimulated neutrophils.

Activated neutrophils represent the main source of myeloperoxidase (MPO), superoxide (SO) and subsequently derived oxygen metabolites. They have important microbicidal activities, however in inflammatory conditions they may secondarily attack surrounding tissues. Overproduction of reactive oxygen species, prolonged or excessive liberation of MPO and other effective yet also toxic substances from neutrophils may participate in disturbed apoptosis, intensify the inflammatory processes and result in serious human diseases. The inhibitory effect of quercetin on PMA stimulated SO generation in isolated human neutrophils was found to be dose-dependent, without affecting the activity of intact isolated neutrophils. At comparable conditions, quercetin was more potent in inhibiting MPO release than SO generation. Our results indicate that quercetin could support resolution of inflammation through decreased activity of neutrophils, i.e. respiratory burst and degranulation.

Naturally appearing N-feruloylserotonin isomers suppress oxidative burst of human neutrophils at the protein kinase C level.

N-feruloylserotonin (N-f-5HT) isomers, isolated from seeds of Leuzea carthamoides (Wild) DC, inhibited dose-dependent oxidative burst in human whole blood and isolated neutrophils in vitro, which were measured by luminol- and/or isoluminol-enhanced chemiluminescence in the following rank order of stimuli: PMA > OpZ > calcium ionophore A23187. In isolated neutrophils that were stimulated with PMA, N-f-5HT isomers were effective against extracellular and intracellular reactive oxygen species. Liberation of ATP, analysis of apoptosis, and recombinant caspase-3 activity revealed that N-f-5HT isomers, used in concentrations up to 100 µM, did not alter the viability and integrity of isolated neutrophils. Western blot analysis documented that in concentrations of 10 and 100 µM, N-f-5HT isomers significantly decreased PMA-induced phosphorylation of PKC α/ß II. The results suggest that N-f-5HT isomers are an effective, naturally occurring substance with a potent pharmacological effect on the oxidative burst of human neutrophils. It should be further investigated for its pharmacological activity against oxidative stress in ischemia-reperfusion, inflammation and other pathological conditions.

Amniopatch - possibility of successful treatment of spontaneous previable rupture of membranes in the second trimester of pregnancy by transabdominal intraamiotic application of platelets and cryoprecipitate.

OBJECTIVE: To outline possibility of successful treatment of spontaneous previable rupture of membranes in the second trimester of pregnancy. INTRODUCTION: Spontaneous previable rupture of membranes (SPROM) in the second trimester of pregnancy is one of the most alarming problems in current obstetrics. Perinatal mortality is about 60 %, one third of which represents intrauterine fetal demise. Surviving neonates suffer from various complications. There are different clinical approaches regarding treatment of SPROM. MATERIAL AND METHODS: We present a case of a 30 year old secundigravida with a history of SPROM at 19+1 weeks gestation. Ultrasonographic examination revealed anhydramnios. Genital cultures and laboratory studies ruled out infectious etiology of SPROM. Due to expected poor neonatal outcome, decision to attempt amniopatch as an experimental therapeutic alternative was made at 21+1 weeks gestation (two weeks after SPROM had occurred). Autologous concentrated platelets followed by autologous cryoprecipitate were administered into the amniotic cavity transabdominally under ultrasound guidance. After 3 days sonographic examination showed normal volume of amniotic fluid. On 22 postoperative day, patient notice some leaking of fluid vaginally. Fetal growth was appropriate, amniotic fluid volume was decreased, however, oligohydramnios never progressed to anhydramnios. Pregnancy ended with primary cesarean delivery at 33+1 weeks gestation. Live born male infant with 1750 g birth weight was delivered. Postnatal development was within normal limits. CONCLUSION: Intraamniotic application of "amniopatch" may represent a possibly successful treatment of spontaneous previable rupture of membranes. This case reports the longest stop of the leaking of amniotic fluid and total prolongation of pregnancy with favorable perinatal outcome after "amniopatch" treatment of spontaneous previable rupture of membranes in the second trimester so far published in available literature.

Suppression of oxidative burst in human neutrophils with the naturally occurring serotonin derivative isomer from Leuzea carthamoides.

OBJECTIVE: Neutrophil leukocytes and macrophages represent professional phagocytic cells. When appropriately stimulated, they undergo dramatic physiological and biochemical changes resulting in phagocytosis, chemotaxis and degranulation with the activation of reactive oxygen species (ROS) production known as the respiratory burst. DESIGN: In this study we analysed the effect of a crystalline complex fraction of four N-feruloyl-serotonin isomers isolated from the seeds of Leuzea carthamoides on the mechanism of oxidative burst of human neutrophils in vitro. RESULTS: N-feruloyl-serotonin (N-f-5HT) inhibited dose-dependently oxidative burst of human whole blood and isolated neutrophils in vitro stimulated with phorbol-myristate-acetate (PMA) as measured by luminol/isoluminol enhanced chemiluminescence.In isolated neutrophils stimulated with PMA, N-f-5HT was effective against extracellular as well as intracellular reactive oxygen species. Western blot analysis documented that N-f-5HT in concentrations of 10 and 100 µM significantly decreased PMA-induced phosphorylation of protein kinase C alpha/beta II. CONCLUSION: The results suggest that N-f-5HT represents an effective naturally occurring substance with potent effect on the oxidative burst of human neutrophils and could be further investigated for its pharmacological activity against oxidative stress in ischaemia-reperfusion, inflammation and other pathological conditions.