Polyacrylamide Disc Gel Research Articles

High Density Lipoprotein Subfraction Changes in End Stage Liver Failure

BACKGROUND: This work was designed to ascertain high-density lipoprotein (HDL) subfractions and HDL-related enzyme alterations in end stage liver failure (ESLF). METHODS: Twenty patients with ESLF and twenty control subjects (liver donors) were chosen for the study. Before the transplant, serum samples from each patient and control were evaluated. Continuous disc polyacrylamide gel electrophoresis was performed to determine HDL subfraction changes and ELISA was carried out to determine serum levels of lecithin–cholesterol acyltransferase (LCAT), apolipoprotein A-1 (ApoA-I) and cholesteryl ester transfer protein (CETP). RESULTS: Liver failure patients had significantly higher levels of triglycerides (TG) and very low-density lipoprotein (VLDL) compared to healthy donors. Additionally, these patients showed significant increases in levels of aspartate aminotransaminase (AST), alkaline phosphatase (ALP), alanine aminotransaminase (ALT), and blood urea nitrogen (BUN), while albumin was notably lower compared to controls. Even though HDL cholesterol concentrations were not considerably altered between the two groups, ESLF patients had a marked raise in the HDL-large subfraction and a significant decline in the HDL-small subfraction in contrast to controls. Moreover, liver failure patients had considerably lower serum ApoA1 levels compared to healthy controls, but there was no significant difference in LCAT and CETP levels between the two groups. CONCLUSION: HDL subfraction profile can distinguish between healthy donors and liver failure patients. The results also indicate that levels of ApoA-1, which performs a crucial function in HDL metabolism, are lower in ESLF. This decrease in HDL-small subfractions may be due to impaired anabolism resulting from hepatic failure.

Sphingolipidomic profile and HDL subfractions in obese dyslipidemic type 2 diabetic patients

PurposeThe aim of the study was to investigate changes in serum sphingolipid levels and high density lipoprotein (HDL) subtypes with relation to low-density lipoprotein cholesterol (LDL-C), non-HDL-C and triglyceride (TG) levels in type 2 diabetes mellitus (T2DM) patients. MethodsBlood was obtained from 60 patients with T2DM. Levels of sphingosine-1-phosphate (S1P), C16-C24 sphingomyelins (SMs), C16-C24 ceramides (CERs), and C16 CER-1 P were determined by LC-MS/MS. Serum concentrations of cholesterol ester transfer protein (CETP), lecithin-cholesterol acyltransferase (LCAT) and apolipoprotein A-1 (apoA-I) were analyzed by enzyme-linked immunosorbent assay (ELISA). HDL subfraction analysis was performed by Disc polyacrylamide gel electrophoresis. ResultsC16 SM, C24 SM, C24-C16 CER and C16 CER-1 P levels were significantly increased in T2DM patients with LDL-C above 160 mg/dL, compared to those with LDL-C below 100 mg/dL. A significant correlation was observed between C24:C16 SM, C24:C16 CER ratios and LDL-C, non HDL-C levels. Higher serum levels of C24 SM, C24-C18 CER and C24:C16 SM ratio was seen in obese T2DM patients (BMI>30) compared to those with BMI 27–30. Patients with fasting TG levels below 150 mg/dL had significantly increased HDL-large and significantly decreased HDL-small fractions compared to those with fasting TG levels above 150 mg/dL. ConclusionObese dyslipidemic T2DM patients had increased levels of serum sphingomyelins, ceramides and HDL-small fractions. The ratio of serum C24:C16 SM, C24:C16 CER and long chain CER levels may be used as diagnostic and prognostic indicators of dyslipidemia in T2DM.

The major yolk protein in sea urchin egg yolk granules is a glycoprotein complex

Major yolk protein (MYP) is the most abundant protein found in the yolk granules of sea urchin eggs. The structure of MYPs stored in the eggs of two sea urchin species from Strongylocentrotus intermedius and Mesocentrotus nudus were investigated. Egg-localized major yolk protein (EGMYP) extracted from the eggs of two species of sea urchins was assessed using disc-polyacrylamide gel electrophoresis and immunoelectrophoresis. The molecular weight of native EGMYP was 595 in S. intermedius eggs and 625 kDa in M. nudus eggs. The use of sodium dodecyl sulfate–polyacrylamide gel electrophoresis and Western blotting analysis under reducing conditions revealed that the EGMYP of S. intermedius separated into four bands (approximately 172, 116, 74 and 68 kDa), while the EGMYP of M. nudus resolved as a set of bands ranging from 175 to 58 kDa (approximately 175, 165, 153, 115, 102, 90, 78, 65 and 58 kDa). Yolk granules from eggs were isolated using sucrose density ultracentrifugation and examined using electron microscopy. The structures of purified EGMYP extracted from eggs and from yolk granules were similar in both species. The EGMYP stored in the eggs of the sea urchins was a glycoprotein complex, the protein structure of which, however, varied between the two species.

Age-dependent variations in the venom proteins of Vipera kaznakovi Nikolsky, 1909 and Vipera ammodytes (Linnaeus, 1758) (Ophidia: Viperidae)

Polyacrylamide disc gel electrophoresis and densitometry analysis were used to analyze venom extracts of Vipera kaznakovi (16.5, 30, and 55 cm) and Vipera ammodytes (28.5, 36.7, and 51.5 cm) specimens of 3 different lengths. V. kaznakovi specimens were collected from Hopa (Artvin Province); 2 V. ammodytes specimens were from Ağva (İstanbul Province) and the longest one was from Yuvacık (Kocaeli Province). The electropherograms of the venom protein samples showed age-dependent qualitative and quantitative variations.

Lipoprotein Subfraction Profile and HDL‐Associated Enzymes in Sickle Cell Disease Patients

Although hypocholesterolemia is a reported finding in sickle cell disease (SCD), low-density lipoprotein (LDL)/high-density lipoprotein (HDL) subfractions and HDL-associated enzymes have not been determined in SCD patients. Blood was collected from 38 hemoglobin (Hb)A volunteers and 45 homozygous HbSS patients who had not received blood transfusions in the last 3 months. Serum lipids were measured by automated analyzer while LDL and HDL subfraction analysis was done by continuous disc polyacrylamide gel electrophoresis. Serum levels of cholesteryl ester transfer protein (CETP), lecithin-cholesterol acyltransferase (LCAT), apolipoprotein B (apoB) and apolipoprotein A-1 (apoA-I) were determined by enzyme-linked immunosorbent assay (ELISA). Total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) levels were significantly decreased, while TG levels were significantly increased in SCD patients compared to controls. A significant decrease in intermediate-density lipoprotein (IDL)-C, IDL-B, IDL-A and LDL-1 fractions were seen in SCD patients, while no significant difference was observed in small dense LDL particles. A significant decrease was seen in HDL-large, HDL-intermediate and HDL-small fractions in SCD patients versus controls. Levels of LCAT and ApoA-1 protein measured in SCD patients were significantly lower while no significant difference was observed in CETP and ApoB protein levels compared to controls. The reduction observed in LDL- and HDL-C in SCD patients was reflected as significantly decreased IDL, LDL-1 and HDL-subfractions. Decreased HDL subfractions may possibly lead to the reduced ApoA-1 and LCAT protein levels observed in SCD patients.

Early Postoperative Changes of HDL Subfraction Profile and HDL-Associated Enzymes After Laparoscopic Sleeve Gastrectomy

This study aimed to determine early postoperative changes of LDL/HDL subfraction profile and HDL-associated enzymes following laparoscopic sleeve gastrectomy (LSG). Thirteen obese patients (mean body mass index (BMI) 52.74 ± 10.97 kg/m(2)) underwent LSG and normal weight control patients (mean BMI 23.56 ± 1.92 kg/m(2)) underwent laparoscopic abdominal surgery. Fasting blood samples were collected prior to surgery, at day 1 after surgery, and after postoperation oral feeding. LDL and HDL subfraction analysis was done by continuous disk polyacrylamide gel electrophoresis. Plasma levels of cholesteryl ester transfer protein (CETP), lecithin-cholesterol acyltransferase (LCAT), and apolipoprotein A-1 (apoA-I) were determined by enzyme-linked immunosorbent assay. Measurement of CETP and LCAT activity was performed via fluorometric analysis. LDL subfraction profile showed no change in both LSG and control group patients. No significant difference was observed in HDL cholesterol, HDL-subfraction distribution, and apoA-I levels in the control group. LSG patients showed a significant increase in HDL-large and a significant decrease in HDL-small fractions at postoperation day 1 compared to preoperation. HDL cholesterol significantly decreased and apoA-I significantly increased in LSG patients after postoperation oral feeding compared to both preoperation and postoperation day 1. Changes in HDL subfraction profile at postoperation day 1 after LSG were accompanied by a significant decrease in CETP protein, LCAT protein, and LCAT activity as compared to preoperation levels. Early changes in HDL subfraction profile and HDL-associated enzymes following LSG suggest that the surgical procedure, irrespective of changes in body weight, affects reverse cholesterol transport.

Effect of insulin analog initiation therapy on LDL/HDL subfraction profile and HDL associated enzymes in type 2 diabetic patients

BackgroundInsulin treatment can lead to good glycemic control and result in improvement of lipid parameters in type 2 diabetic patients. This study was designed to evaluate the effect of insulin analog initiation therapy on low-density lipoprotein (LDL)/ high-density lipoprotein (HDL) sub-fractions and HDL associated enzymes in type 2 diabetic patients during early phase.MethodsTwenty four type 2 diabetic patients with glycosylated hemoglobin (HbA1c) levels above 10% despite ongoing combination therapy with sulphonylurea and metformin were selected. Former treatment regimen was continued for the first day followed by substitution of sulphonylurea therapy with different insulin analogs (0.4 U/kg/day) plus metformin. Glycemic profiles were determined over 72 hours by continuous glucose monitoring system (CGMS) and blood samples were obtained from all patients at 24 and 72 hours. Plasma levels of cholesteryl ester transfer protein (CETP), lecithin-cholesterol acyltransferase (LCAT), apolipoprotein B (apoB) and apolipoprotein A-1 (apoA-I) were determined by enzyme-linked immunosorbent assay (ELISA). Measurement of CETP and LCAT activity was performed via fluorometric analysis. Paraoxonase (PON1) enzyme activity was assessed from the rate of enzymatic hydrolysis of phenyl acetate to phenol formation. LDL and HDL subfraction analysis was done by continuous disc polyacrylamide gel electrophoresis.ResultsMean blood glucose, total cholesterol (TC), triglyceride (TG) and very low-density lipoprotein cholesterol (VLDL-C) levels were significantly decreased while HDL-C levels were significantly increased after insulin treatment. Although LDL-C levels were not significantly different before and after insulin initiation therapy a significant increase in LDL-1 subgroup and a significant reduction in atherogenic LDL-3 and LDL-4 subgroups were observed. Insulin analog initiation therapy caused a significant increase in HDL-large, HDL- intermediate and a significant reduction in HDL-small subfractions. CETP protein level and activity was significantly increased while apoB levels were significantly decreased following insulin analog initiation therapy. No significant difference was found in LCAT mass, LCAT activity, apoA-I and PON-1 arylesterase levels following insulin initiation therapy.ConclusionThese findings indicate that insulin analog initiation therapy activates lipid metabolism via up-regulating CETP and shows anti-atherogenic effects by increasing HDL-large and decreasing LDL-3 and LDL-4 subfractions in a short time period.

Increased Small Dense LDL and Decreased Paraoxonase Enzyme Activity Reveals Formation of an Atherogenic Risk in Streptozotocin-Induced Diabetic Guinea Pigs

This study aimed to investigate LDL subfraction distribution as well as serum cholesteryl ester transfer protein (CETP), lecithin-cholesterol acyltransferase (LCAT), and paraoxonase (PON1) activity in streptozotocin-induced diabetic guinea pigs. Materials/Methods. Guinea pigs were given a single intraperitoneal (ip) injection of streptozotocin (STZ) and animals having fasting blood glucose levels greater than 200 mg/dl, were considered diabetic. Protein levels of LCAT and CETP were determined via ELISA. Paraoxonase activity was measured kinetically by the formation of phenol while LDL subfraction analysis was done by disc polyacrylamide gel electrophoresis. Results. Plasma glucose and high-density lipoprotein (HDL) cholesterol were significantly increased while total cholesterol and LDL cholesterol were significantly decreased in diabetic guinea pigs compared to controls. LDL subfraction analysis revealed a significant decrease in nonatherogenic LDL-2 subfraction and a significant increase in atherogenic LDL-4 subfraction in diabetic guinea pigs compared to controls. Plasma CETP and PON1 levels were significantly decreased while LCAT showed no significant difference in diabetic guinea pigs compared to controls. Conclusion. Decreased non-atherogenic LDL-1, LDL-2 subfractions, increased small dense LDL-4 subfraction, and decreased PON1 activity, reveals formation of an atherogenic risk in diabetic guinea pigs. Decrease in CETP levels supports the observed increase in HDL cholesterol levels in diabetic guinea pigs.

PREPARATION, PURIFICATION AND PROPERTIES OF LIPASE FROM HEPATOPANCREAS OF TRA (PANGASIUS) CATFISH

Lipase from the hepatopancreas of Tra (Pangasius) catfish was purified by ammonium sulfate fractionation, followed by ion-exhange chromatography on DEAE Cellulose and gel filtration Sephadex G-75. The preparation was homogeneous on polyacrylamide disc gel electrophoresis. The specific activity of the purified enzyme was 37.95 times higher than that of the crude extract. The enzyme showed a molecular weight of 57000 Da. The pH and temperature optima of purified lipase were 8 and 500C respectively. Enzyme activity was enhanced by Ca2+ but inhibited by heavy metals Zn2+, Cd2+, Mg2+.

Haptoglobin polymorphism among the tribal groups of southern Gujarat.

BACKGROUND:Gujarat is located at the western most point of the Indian subcontinent. Valsad and Surat districts are part of the ‘tribal belt’of Gujarat and constitute 29.1% of total tribal population of Gujarat. These tribal populations are a rich source of gaining insights in the patterns of genetic diversity and genetico-environmental disorders against the back drop of their ecological, historical and ethnographic aspects.AIM:The objectives were to find out a) the genetic diversity among the tribes of Gujarat with reference to haptoglobin (Hp) locus b) the relationship between Hp polymorphism and sickle cell anemia/trait.MATERIALS AND METHODS:431 individuals belonging to eight tribal groups were studied for Hp polymorphism using polyacrylamide disc gel electrophoresis (PAGE). Hb*S was screened by dithionate tube turbididy (DTT) test and confirmed using cellulose acetate membrane electrophoresis (CAME).STATISTICAL ANALYSIS:Allele frequency was calculated by direct gene counting method. Average heterozygosity and gene diversity were computed using software DISPAN. Analysis of molecular variance (AMOVA) was estimated using software ARLEQUIN version 3.1.RESULTS AND CONCLUSIONS:Pattern of allele frequency distribution showed preponderance of Hp2 allele in all the eight tribal groups, which is in accordance with its frequency in different populations of Indian subcontinent. Total average heterozygosity (HT) was found to be low (0.160) but the level of genetic differentiation (GST) was found to be moderately high (5.6%). AMOVA analysis indicated least among group variance between west and south Indian populations (-0.04%) indicating the affinities of the tribes of Gujarat with that of Dravidian speaking groups. Analysis of Hp phenotypes among sickle cell anemia/ trait individuals revealed a high frequency of Hp 0-0 phenotype (92.7%) among SS individuals as opposed to only 9.7% among AS individuals, reaffirming the selective advantage of HbAS state in relation to hemolytic disorders.

Electrophoretic Comparison of Blood-serum Proteins of Apathya cappadocica (Sauria, Lacertidae) Subspecies from Anatolia

Blood-serum proteins of the known subspecies of Apathya cappadocica (Werner, 1902) were studied comparatively by polyacrylamide disc gel electrophoresis. In order to obtain useful biochemical data for classification, differences between the electrophoreograms of the samples included in the morphologically different subspe - cies were distinguished qualitatively and quantitatively. These comparisons indicated that electrophoretic results supported morphological discrimination of the known subspecies of A. cappadocica.

A partially degraded form of human plasminogen in circulating blood

Summary A method for separate measurement of the amounts of the native form and partially degraded form of plasminogen and plasmin in human circulating blood was devised based on their different mobilities on disc polyacrylamide gel electrophoresis. The relative amounts in the circulating blood as estimated by the method were as follows: 78.5% native form, 6.1% partially degraded form and 15.5% plasmin, in the resting state of healthy adults. The partially degraded form increased to about 50% after strenuous exercise load. The present study showed that relatively large amounts of the partially degraded form can be produced in the circulating blood, even though abundant plasmin inhibitors exist there. These data suggest that the partially degraded form may play an important role in thrombolysis in vivo.

A partially degraded form of human plasminogen in circulating blood

A method for separate measurement of the amounts of the native form and partially degraded form of plasminogen and plasmin in human circulating blood was devised based on their different mobilities on disc polyacrylamide gel electrophoresis. The relative amounts in the circulating blood as estimated by the method were as follows: 78.5% native form, 6.1% partially degraded form and 15.5% plasmin, in the resting state of healthy adults. The partially degraded form increased to about 50% after strenuous exercise load. The present study showed that relatively large amounts of the partially degraded form can be produced in the circulating blood, even though abundant plasmin inhibitors exist there. These data suggest that the partially degraded form may play an important role in thrombolysis in vivo.

Isolation and Characterization of a Novel Acid Proteinase, Tropiase from Candida tropicalis IFO 0589

A novel acid proteinase (Tropiase) was isolated from Candida tropicalis IFO 0589 by DE52-cellulose, and DEAE-Cosmogel column chromatographies. The purified tropiase gave a single band on disc polyacrylamide gel electrophoresis, isoelectric focusing and sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. The enzyme preparation had a molecular weight of 23,900, isoelectric point of pH 5.1, optimum pH range of 7 to 9 and possessed 208 amino acid residues. The enzyme hydrolyzed casein, fibrinogen, keratin and collagen. The purified tropiase demonstrated hemorrhagic and capillary permeability-increasing activities. Inhibition of tropiase occurred with leupeptin and N-bromosuccinimide, however, no inhibition was observed with alpha(2)-macroglobulin, soybean trypsin inhibitor, benzamidine-HCl or diisopropyl fluorophosphate.

Effects of long wave ultraviolet radiation on photosensitizing and related compounds. I. The effects on the fluorescence of topically applied compounds.

SUMMARY The effects of long wave ultraviolet radiation (320–400 nm) on the binding of twenty-five fluorescent photosensitizing and related compounds to the major saline soluble guinea-pig epidermal proteins were studied. Binding before or after irradiation was detected on polyacrylamide disc gel electrophoresis by correlating the fluorescent migration pattern with the amido black counterstained pattern. Effects were seen only on photo-allergic and structurally related compounds (phenothiazines and halogenated salicylanilides). The binding of five phenothiazine derivatives was detected only after irradiation. Ten of twelve halogenated salicylanilides were bound without irradiation, and eleven were bound following it. Ultraviolet exposure appeared to increase the amount of each halogenated salicylanilide that was bound and/or to alter the existing complex.

Acarbose ameliorates atherogenecity of low-density lipoprotein in patients with impaired glucose tolerance

Acarbose, an α-glucosidase inhibitor, is administered to control blood glucose levels. The drug also reduces the risk of cardiovascular disease, but the underlying mechanism is still to be elucidated. We therefore hypothesized that treatment with acarbose ameliorates the atherogenecity of low-density lipoprotein (LDL), a key molecule in atherogenesis. Patients with impaired glucose tolerance were or were not treated with acarbose (acarbose-treated group [n = 20] and control group [n = 20], respectively) for 3 months under dietary therapy. The oxidative susceptibility of LDL was determined by measuring lag time for the formation of dienes in the presence of CuSO 4. The lag time was significantly longer in the acarbose-treated group than in the control group before treatment. Moreover, the density gradient lipoprotein separation and disk polyacrylamide gel electrophoresis analyses showed that acarbose reduced the amount of small dense LDL, a more atherogenic and oxidatively susceptible form of LDL. We also found that the fatty acid composition of LDL changed after the treatment: polyunsaturated ( ω-3) fatty acid, a beneficial substance for preventing cardiovascular disease, was significantly increased, whereas saturated fatty acids and triglyceride were decreased in the LDL of the acarbose-treated group. The present findings suggest that acarbose treatment reduces the risk of cardiovascular diseases by ameliorating the atherogenecity of LDL.

Isolation and Characterization of a Novel Elastase Inhibitor, AFLEI from Aspergillus flavus

A novel elastase inhibitor from Aspergillus flavus (AFLEI) was isolated, and biochemical properties of AFLEI were examined. Column chromatography using diethylaminoethyl (DE) 52-Cellulose and Sephadex G-75 was used to purify the inhibitor. The final preparation was found to be homogeneous as indicated by a single band after disc polyacrylamide gel (PAGE) and isoelectric focusing electrophoreses. AFLEI had a molecular weight of 7,525.8 as determined by TOF-MS (time of flight mass spectrometry). The elastolytic activity of elastases from A. flavus, A. fumigatus and human leukocytes were inhibited by AFLEI. However, this activity from porcine pancreas elastase, trypsin, chymotrypsin, thrombin, and Ac1-Proteinase from snake venom was not affected by AFLEI. The fibrinogenase activity of the elastase from A. flavus was inhibited by AFLEI. AFLEI was inhibited by alpha2-macroglobulin. However, ethylenediaminetetraacetic acid (EDTA-2Na), benzamidine, chymostatin, tosyl phenylalanine chloromethyl ketone (TPCK) and dithiothreitol (DTT) did not show any inhibitory effect on the elastase inhibitory activity of AFLEI.

Changes of Serum Alkaline Phosphatase Activity in Dry and Lactational Cows

Serum alkaline phosphatase (ALP) activities were measured during dry and lactational periods to investigate the influence of lactation on serum ALP activity in cows. Higher levels of serum ALP activity were seen in lactational periods than in dry periods. The serum activities of bone-specific ALP (BALP), liver ALP (LALP), tartrate resistant acid phosphatase (TRAP) and aspartate aminotransferase also increased in lactational periods. ALP activities in the bone extract and in whey were decreased at similar rates by the addition of lectin. Moreover, since the ALP band in whey was observed to have the same migration in polyacrylamide gel (PAG) disk electrophoresis as that of the bone extract, analysis of ALP isoenzymes by lectin affinity or PAG disk electrophoresis could not distinguish ALP originating from the mammary gland from that of bone. In this study, it was clear that the increased level of serum ALP activity was due to increases of BALP and LALP in lactational periods. However, the extent of the influence of ALP originating from the mammary glands on serum ALP activity was unknown. Judging from changes of BALP and TRAP activities in the serum and the correlation between the both, it was guessed that ALP originating from the mammary glands influenced serum ALP activity.

KMP016 Molecular biological analysis of Interferon resistance in hepatitis C virus genotype 1b

An elongation factor with activity corresponding to that of mammalian EF-2 (avian EF-2) has been purified 500-fold from hen oviduct to apparent homogeneity using a 6-step procedure. The homogeneity of the preparation has been determined by the following criteria: (1) a single band on three polyacrylamide disc gel electrophoretic systems (anodic, cathodic, and sodium dodecyl sulfate); (2) velocity sedimentation ultracentrifugation; and (3) amino terminal analysis. These studies show that purified avian EF-2 is a single polypeptide. The purified protein has been found to have activity in the following reactions: (1) synthesis of polyphenylalanine from [14C]Phe-tRNA in the presence of rabbit reticulocyte EF-1; (2) formation and release of the dipeptide[14C]phenylalanylpuromycin also in the presence of EF-1; (3) the ribosome-dependent hydrolysis of [γ-32P]GTP; and (4) the diphtheria toxin-dependent [3H]ADP-ribose incorporation from [3H]NAD+. The activity of the factor in polypeptide synthesis is inhibited by fusidic acid and ADP-ribosylation. Avian EF-2 was inactive in stimulating polyphenylalanine synthesis from [14C]Phe-tRNA in a protein synthesis system derived from Escherichia coli.The molecular weight was determined to be 80 000 by band sedimentation ultracentrifugation, 96 000 by Sephadex G-200 gel filtration chromatography and 93 000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The Stokes radius was estimated to be 37.5 Å. The calculated axial ratio of the molecule was 4. The sedimentation coefficient (s20,w) was 4.92 S at infinite dilution. Comparison of the amino acid composition analysis of avian EF-2 with other mammalian EF-2 molecules as well as EF-G from E. coli showed that there was a striking degree of similarity. The molecular weight calculated from the amino acid residue composition was 81 500. The pI was estimated to be 6.75 by the technique of zone convection electrofocusing. In addition fluorescence and circular dichroism spectra of purified avian EF-2 were analyzed. Circular dichroism spectrum analysis estimated the α-helix content of the molecule to be 19–20%.

Low-density lipoprotein subfractions and cardiovascular risk in hypertension: relationship to endothelial dysfunction and effects of treatment.

Although hypertensive patients are at particular risk of vascular complications, the possible contribution of an atherogenic lipoprotein profile and endothelial dysfunction to this risk is unclear. We investigated this by measuring LDL subfractions and flow-mediated dilation (FMD) (reflecting endothelial dysfunction) in a cohort of high-risk hypertensive patients. We studied 84 hypertensive patients (74 men; mean age, 64 years; SD 8). Chylomicron-free LDL subfractions were analyzed by disc polyacrylamide gel electrophoresis, producing an LDL score, with higher scores being equivalent to a greater proportion of the more atherogenic LDL subfractions. High-resolution ultrasound was used to assess endothelium-dependent brachial artery FMD after reactive hyperemia after vessel occlusion. Baseline levels were compared with 61 age- and gender-matched healthy normotensive control subjects. Mean LDL score was higher and FMD impaired in hypertensive subjects compared with control subjects. These indexes were significantly improved after 6 months of cardiovascular risk factor management. LDL score correlated significantly with the 10-year Framingham coronary heart disease risk score, with a negative correlation with FMD (both P<0.001). Abnormal atherogenesis and endothelial dysfunction are both present in hypertension and appear to be related to each other, potentially leading to vascular complications. The abnormal LDL scores also correlate with the 10-year cardiovascular risk and can be positively influenced by cardiovascular risk management.