BACKGROUND

Low doses of androgen are used in women for the symptomatic treatment of sexual dysfunction and premenstrual syndrome (PMS). However, little is known about the long-term safety of androgen use in women. This study investigated the effects of low dose exogenous testosterone (T) on lipid metabolism, markers of activation of the coagulation system and ultrasonographic ovarian morphology in women.

METHODS

Twenty-two patients with severe PMS (age 39.6 +/- 3.1 years, mean +/- SD) treated with subcutaneous T implants (100 mg six monthly) for at least two years (mean duration 3.3 (+/- 0.9 years) were compared with 22 age-matched (age 37.7 +/- 2.9 years) control patients with severe PMS who had not previously received T treatment. All women continued to have regular menses.

METHODS

Fasting blood samples were obtained for measurement of lipids and clotting factors and ovarian ultrasound examination carried out between days 1-4 of the menstrual cycle (2.3 +/- 1.2 months after the T implant in T-treated group).

RESULTS

Mean plasma T was 4.5 +/- 2.2 nmol/l, and 1.9 +/- 0.6 nmol/l in the treated and control groups, respectively. In the T-treated group apolipoprotein-A1 (Apo-A1) (treated 99.2 +/- 12 vs controls 116.2 +/- 27.7 g/l, P < 0.01) and high density lipoprotein cholesterol (HDL-C) (treated 1.3 +/- 0.3 vs controls 1.5 +/- 0.4 nmol/l, P < 0.01) were significantly decreased. In addition very low density lipoprotein cholesterol (VLDL-C) (treated 0.4 +/- 0.3 vs controls 0.2 +/- 0.1 nmol/l, P < 0.05) was increased in T-treated patients. There were no differences in total serum cholesterol and triglyceride or low density lipoprotein cholesterol (LDL-C), apolipoprotein B (Apo-B), lipoprotein(a), lecithin:cholesterol acyltransferase and cholesteryl ester transfer protein activity. There was no difference in clotting factors between the two groups which included prothrombin time, fibrinogen, antithrombin-III, protein-C, protein-S (total and free), tissue plasminogen activator, plasminogen activator inhibitor, beta-thromboglobulin and prothrombin fragments 1.2. Ultrasound showed normal ovarian architecture with no evidence of polycystic ovarian changes in any patients in the T-treated group. No patient experienced adverse symptoms while on T treatment, in particular, there were no complaints of hirsutism or acne and no one requested termination of treatment.

CONCLUSIONS

Low-dose testosterome administration to women for over two years did not induce changes in ovarian architecture but had small, potentially atherogenic effects on some parameters of lipid and lipoprotein metabolism. However, no differences were detected in markers of activation of the clotting system to indicate an actual increase in the risk of thrombosis. Overall, this study provides largely reassuring data about the safety of low-dose androgen treatment in women. However, caution should be exercised in women with existing or a familial predisposition to lipid abnormalities, because of the small but significant changes found in HDL-C, apo-A1 and VLDL-C.

Progressive accumulation of amyloid-β peptide (Aβ) in the brain is implicated as the central event in the development of Alzheimer's disease (AD). It is thought that extracellular Aβ triggers toxic signals leading to neurodegeneration. The events downstream of Aβ however are not entirely clear. Clusterin (Apo J) is one of the major risk factors for sporadic form of AD. Clusterin binds to Aβ and prevents Aβ aggregation. In addition, clusterin promotes Aβ degradation and accelerates Aβ clearance from the brain. Clusterin thus protects neurons from Aβ and loss of clusterin level in the brain is implicated as promoting AD pathology. In this study, we found that the level of clusterin protein but not mRNA is reduced in the brains of 3xTg-AD mice. When rat hippocampal primary neurons were treated with Aβ1-42, level of clusterin protein but not mRNA was downregulated. Aβ1-42-induced downregulation of clusterin was blocked by lysosome inhibitors bafilomycin A1 and ammonium chloride. In neurons, Aβ1-42 induced expression of sortilin, a lysosomal sorting protein that targets proteins to lysosome for degradation. In BE(2) M17 human neuroblastoma cells, clusterin bound to sortilin and when sortilin expression was silenced, Aβ1-42-induced clusterin downregulation was almost completely blocked. Our data demonstrate that in neurons, Aβ1-42 promotes lysosomal degradation of clusterin by inducing expression of sortilin and provide a novel mechanism by which Aβ promotes AD pathogenesis.

We analyzed correlations between apolipoprotein B (apo B), cholesterol and phospholipids (preponderant lipids) in low-density lipoproteins (LDL) as well as between apolipoprotein A1 (apo A1) and these same lipids in high-density lipoproteins (HDL), during the acute phase of myocardial infarction. In LDL, a very elevated and stable correlation (r) was observed between these parameters, and the coefficients of regression (b) did not differ significantly during the period studied. In HDL, there was a decrease in r and b values from day 1 to day 2, then an increase after day 2. We hypothesize that these disturbances in HDL composition may be due to a greater endocytosis of LDL at day 2, leading to intracellular increase in cholesterol and phospholipids. Part of these lipids could be taken up by HDL molecules, causing a transient overload.

Plasma proteins provide precise information about the physiological status of an individual. In this study, we compared the plasma protein profiles of 168 individuals from the Adélé ethnic group, from an isolated rural area of Togo, with those of 159 individuals from an urban population from the capital, Lomé. The Adélé villages are located in the Atakora mountains. The subjects were volunteers, all apparently healthy and aged between 18 and 65 years. We separated serum proteins by electrophoresis and identified proteins specific for nutritional, inflammatory and immune status. The Adélé significantly higher total serum protein concentrations than the urban individuals, with higher concentrations of a1 globulins (2.35 +/- 0.57 g/L versus 1.94 +/- 0.52 g/L) and g globulins (22.19 +/- 5.67 g/L versus 16.98 +/- 5.23 g/L) and lower concentrations of b globulins (6.83 +/- 1.56 g/L versus 7.34 +/- 1. 52 g/L). The Adélé also had lower plasma concentrations of albumin (41.91 +/- 5.74 g/L versus 44.56 +/- 6.32 g/L), tranferrin (2.5 +/- 0.52 g/L versus 3.03 +/- 0.6 g/L), haptoglobin (0.57 +/- 0.59 g/L versus 1.32 +/- 0.89 g/L) and IgA (2.3 +/- 0.89 g/L versus 2.88 +/- 1.12 g/L) and higher plasma concentrations of orosomucoid (0.85 +/- 0.26 g/L versus 0.69 +/- 0.27 g/L); IgG (25.3 +/- 7.11 g/L versus 21. 79 +/- 6.5 g/L) and IgM (4.25 +/- 2.83 g/L versus 2.25 +/- 1.0 g/L). The data obtained for the Adélé and urban populations were similar to those obtained for European populations except for IgM (higher in the Adélé than in the urban and European populations), IgG and CRP (higher for the Adélé and urban populations than for European populations). Nutritional status, as estimated by albumin and transferrin concentrations, was higher in the urban population of Lomé than in the Adélé population but the Adélé population suffered no malnutrition problems. These results are consistent with those of a previous study, using apo A-I concentrations as an index of nutritional status. Apo A-I has also been shown to be a reliable indicator of nutritional status, as prealbumin concentration alone is sufficient for the early diagnosis of protein malnutrition. The very high concentrations of plasma CRP obtained indicate the presence of an inflammatory syndrome in the Adélé and urban populations, as this protein is the first acute phase protein to be detected. However, the orosomucoid concentrations obtain-ed provide no evidence of significant inflammation. The high affinity of haptoglobin (Hp) for hemoglobin (Hb) results in the formation of soluble Hp-Hb complexes, reducing the value of Hp as a marker of the acute phase of inflammation. The frequency os sickle cell disease was higher in the Adélé population than in the urban population (10-25% versus 2-6%). Hemoglobinopathies are correlated with haptoglobin concentration and thus plasma haptoglobin concentration was lower in the Adélé population than in the urban population. The plasma concentrations of a1-antitrypsin in this study were similar to those reported for Europeans. The plasma concentration of protease inhibitors, such as a1-antitrypsin, increased as protease levels increased. These data confirm that the Adélé and urban populations suffer no disease due to high levels of protease release into the bloodstream. They also show that a1-antitrypsin is of some value as an acute phase marker protein. The acute nature of the inflammatory syndrome (as assessed by CRP concentration) in the Adélé and urban populations was confirmed by the hyperglobulinemia (high levels of production of IgM and IgG antibodies) observed in these populations. The Adélé and Lomé urban populations live in a tropical environment in which they are continuously in contact with infectious agents. This results in repeated stimulation of the immune system in both these populations. This study of plasma proteins in the Adélé provides insight into the physiological conditions of this ethnic group, w

Statin and aspirin form the therapeutic cornerstone in patients with coronary artery disease (CAD) and diabetes. Little is known about relationship of statins with blood thrombogenicity and inflammation in these patients.

Two hundred nine consecutive patients with diabetes and suspected CAD undergoing elective cardiac catheterization were divided in groups based on statin treatment in the Multi-Analyte, Thrombogenic, and Genetic Markers Atherosclerosis study. Urinary 11-dehydrothromboxane B2 (11-dh-TxB2), lipid profile and oxLDL/β2GPI were measured by AspirinWorks™ ELISA assay, vertical density gradient ultracentrifugation and immunoassay respectively. Thrombelastography, and ADP- and collagen-induced light transmittance aggregometry assessed thrombogenicity. CAD was classified as none/minor [<20% diameter stenosis (DS)], moderate (20-75% DS), or severe (>75% DS).

Severe, moderate, and no CAD was observed in 66, 19, and 15% of patients respectively. Patients on statins had significantly lower 11-dh-TxB2, collagen-induced aggregation, total cholesterol, total LDL, LDL3, oxidized-LDL, Apo B100, and ApoB100/A1 ratio (p<0.01 for all). Statin therapy demonstrated a lower proportion of patients with high urinary 11-dh-TxB2 (>1500pg 11-dh-TxB2/mg creatinine) (25 vs. 57%, p=0.01).

Statins along with aspirin, confers additional anti-inflammatory and antithrombotic effect in diabetics with CAD. Urinary 11-dh-TxB2 may be a useful biomarker for personalizing statin therapy.

BACKGROUND

There are conflicting reports about the effects of angiotensin converting enzyme inhibitors on insulin sensitivity and glycaemic control. In addition, the chronic effects of ACEI on insulin sensitivity in normotensive but insulin resistant individuals have been controversial.

OBJECTIVE

To determine the long-term effects of low-dose captopril or enalapril on insulin sensitivity and lipid parameters in normotensive non-insulin dependent diabetic volunteers.

METHODS

Twenty-eight normotensive non-insulin dependent diabetes mellitus subjects on diet alone or diet plus oral hypoglycaemic agents were randomized in a single-blind cross-over study to receive either captopril (12.5 mg daily) or enalapril (5 mg daily). Initially, captopril was compared with enalapril for 28 days with a 28-day washout period between drug regimens. For the long-term study, the subjects then remained on the second ACEI for a further 11 months. Insulin sensitivity was measured using the isoglycaemic hyperinsulinaemic clamp (insulin infusion rate 20 mIU/kg/min) at the start and completion of each part of the cross-over study and then at 3, 6 and 12 months of drug therapy. Fasting glucose, insulin, HbA1, lipids and lipoproteins were measured at the start of each clamp.

RESULTS

No first or second order carry-over effects were demonstrated between the ACEIs. No differences were detected between enalapril and captopril on insulin sensitivity at any of the time points. Statistically significant hypotension was avoided, and at doses used the ACEIs did not modify any parameters of glycaemic control over the 12-month study period. There were no significant alterations in plasma cholesterol, triglycerides, HDL cholesterol or Apo A1 levels during the study.

CONCLUSIONS

Long-term low-dose ACEIs (captopril/enalapril) do not modify insulin sensitivity, glycaemic control or lipids in normotensive non-insulin dependent diabetic subjects.

An abnormal plasma lipid and lipoprotein profile is an independent and strong predictor of mortality and morbidity from coronary artery disease (CAD). We report on plasma lipid and lipoprotein profiles with respect to race, age, obesity, blood pressure (BP), smoking, and drinking history in 1,292 male veterans with a diastolic BP of 95 to 109 mm Hg while off antihypertensive medications. Blacks had 24% (p <0.001) lower triglycerides than whites. In contrast, the following parameters were higher in blacks than in whites by the indicated percentages: high-density lipoprotein (HDL) cholesterol, 16% (p <0.001); HDL2 cholesterol, 36% (p <0.001); apolipoprotein (Apo) A1, 8% (p <0.001); HDL/low-density lipoprotein (LDL), 18% (p = 0.018); HDL2/LDL, 36% (p = 0.031); HDL2/HDL3, 21% (p <0.001); and Apo A1/Apo B, 15% (p <0.001). Triglycerides were unchanged up to age 60, but were lower by 24% (p <0.001) in those aged>> or = 70. Apo A1 levels were higher (p <0.001), whereas LDL cholesterol was lower (p <0.008) in moderate alcohol consumers versus abstainers. Triglycerides were higher (p <0.001), whereas HDL, HDL2 cholesterol, and Apo A1 were lower (p <0.001) with increasing obesity. Moderate alcohol consumption had a strong favorable effect on HDL, HDL2, and HDL3 cholesterol among subjects of normal weight, but this effect was diminished in obese subjects. Total and LDL cholesterol were higher by 6.4% (p = 0.001) and 9.4% (p <0.003), respectively, whereas HDL cholesterol remained unchanged in those with diastolic BP of 105 to 109 mm Hg versus those with diastolic BP of 95 to 99 mm Hg. We conclude that hypertensive black men have lipid and lipoprotein profiles indicative of less CAD risk than white men. Chronic moderate alcohol consumption correlates with a favorable plasma lipid and lipoprotein profile in normal, but not obese, men. Obesity is associated with an adverse plasma lipid and lipoprotein profile. Thus, race, alcohol intake, and obesity may be important modifiers of CAD in untreated hypertensive men.

A pharmacological study was carried out in a group of 20 subjects composed of 13 coronary patients and 7 normal close relatives (first degree) who were considered at a high risk for coronary heart disease (CHD) because of their low levels of high density lipoprotein cholesterol (HDLch) (mean +/- SD: 34.1 +/- 5.2 mg/dl) and their high total cholesterol/HDLch (Tch/HDLch) ratio (mean +/- SD:6.7 +/- 1.1), despite their normal serum lipid values. With the purpose of normalizing these parameters they were submitted to a 4-month treatment with bezafibrate, a hypolipidemic agent which has a known effect in increasing HDLch and in decreasing the Tch/HDLch ratio. At the end of the study total serum cholesterol and triglycerides decreased significantly by 10 and 30%, respectively (p less than 0.01). HDL increased in its cholesterol content by 33% reaching a value of 45.4 +/- 9.8 mg/dl (mean +/- SD, p less than 0.01) as well as in its apolipoprotein A1 (Apo A1) content (13%, p less than 0.02). HDL2 subfraction also rose in cholesterol and in Apo A1: by 90 and 38%, respectively (p less than 0.01). HDL3 subfraction rised only its cholesterol content by 24% (p less than 0.01). Tch/HDLch ratio was significantly reduced (p less than 0.01) to a value of 4.6 +/- 0.9 (mean +/- SD). Total serum Apo B diminished by 14% (p less than 0.01). No adverse effects were observed during the follow-up.(ABSTRACT TRUNCATED AT 250 WORDS)

Without appropriate interventions, prediabetes is typically followed by type II diabetes. Eggs are a rich source of important nutrients including protein, vitamins, minerals, carotenoids and lecithin. In this 12-week, parallel, randomized controlled trial, 42 overweight or obese individuals between the ages of 40 and 75 years with pre- and type II-diabetes were included. Participants were randomly assigned to receive either one large egg per day or an equivalent amount of egg substitute for 12 weeks. Blood samples were obtained to analyze lipid profile and biomarkers associated with glycemic control at all time points. Regular egg consumption resulted in improvements of fasting blood glucose, which was significantly (P = 0.05) reduced by 4.4% at the final visit in the egg group. Participants in the egg group had significantly (P = 0.01) lower levels of homeostatic model assessment of insulin resistance (HOMA-IR) at all visits. In the egg group, ATP-binding cassette protein family A1 (ABCA1) was significantly higher at the 6-week visit (0.78 ± 0.21 vs. 0.28 ± 0.05 mg dL-1, P < 0.001) and tended to be higher at the final visit (0.62 ± 0.11 vs. 0.55 ± 0.18 mg dL-1, P = 0.1). The mean apolipoprotein A1 (apo A1) level was also significantly higher at the final visit in the egg group compared to the control (147.43 ± 5.34 vs. 142.81 ± 5.09 mg dL-1, P = 0.01). There were no significant changes in total cholesterol and low-density lipoprotein cholesterol (LDL-C) levels. Daily consumption of one large egg may reduce the risk of diabetes without having any adverse effects on lipid profiles in individuals with pre- and type II diabetes.

BACKGROUND

In the previous studies, fasting insulin and apo-lipoproteins are considered as one of the risk-factor of coronary artery disease (CAD) but did not have the same results. In this study, we attempted to define the association of high fasting insulin and apo-lipoproteins of serum in non-diabetic patients who were afflicted with coronary arteries disease with severity of coronary arteries involvement.

METHODS

This study was conducted between September 2011 and February 2012 on three groups, each one consisting of 100 members while using angiographic scores of Gensini with three equal groups with low, medium, and high stenosis of coronary arteries. The evaluation of non-diabetic patients afflicted with CADs, included the fasting glucose level less than 126 mg/dl or non-consumption of blood glucose reduction drugs or negativity history of diabetes.

RESULTS

In this study, there were 300 non-diabetic patients afflicted with CAD in three groups of low, medium, and high extremity. Due to attained results, the patients afflicted with high CAD had a higher level of insulin (18.3 ± 0.8) in relation with low and medium groups (P < 0.001). As it was observed, the level of serum apo-lipoproteins of A1 (APO-A1) in low group of CAD (175 ± 36.4) is meaningfully higher than its quantity in high-CAD group (158 ± 42.4, P < 0.001). Furthermore, the quantity of serum apo-lipoproteins of B (APO-B) in mild CAD group (139 ± 30.4) is meaningfully less than severe CAD group (155.21 ± 29.7, P < 0.001).

CONCLUSIONS

Our findings show that insulin, APO-A1, APO-B, and total cholesterol measurement is a good case for defining the severity of coronary artery involvement, while high-density lipoprotein, low-density lipoprotein, and triglyceride are not important risk-factors.

The relationship between erythrocyte sodium-lithium counter-transport activity, serum insulin, lipids and demographic factors was examined in 93 normoglycaemic predominantly normotensive individuals with mild fasting hypercholesterolaemia (greater than 5.2 mmol/l). The major significant univariate correlates of sodium-lithium counter-transport activity were fasting serum triglycerides, HDL cholesterol, the ratio of fasting glucose: insulin, apo A1, alcohol consumption and apo B. Stepwise multiple regression analysis revealed 24% of the variability in sodium-lithium counter-transport activity could be accounted for by independent contributions of fasting serum triglycerides, alcohol consumption, the fasting glucose/insulin ratio and apo A1 and ANOVA confirmed a significant relationship with fasting insulin measures that was independent of serum triglycerides (P less than 0.05). The relationship between erythrocyte sodium-lithium counter-transport activity and concentrations of serum triglycerides, HDL components, insulin and additionally alcohol consumption, could reflect the influence of those variables on erythrocyte structure and function.

The Göttingen Risk-, Incidence- and Prevalence Study (abbreviated GRIPS) consists of several epidemiological projects, among which project B is currently the most important one. Project B is a prospective cohort study, which was started in 1982 by a baseline investigation, including approximately 6000 men aged 40 to 60 years. Follow-up investigations were performed in 1985 and 1987 in order to record reasons of death and incidences of newly developed diseases. Further follow-up investigations are planned for 1990 and 1992. Until now incidence data of death and diseases are available for a 5-year observation period (January 1982-December 1986), and for more than 95% of the original study participants. The results clearly indicate that plasma levels of LDL-cholesterol, followed by plasma concentrations of total cholesterol and apoprotein B are the predominant predictors for the risk of coronary artery diseases (CAD; i.e. fatal and non fatal myocardial infarction as well as sudden coronary death and chronic coronary heart disease). Further significantly positive associations to the incidence of myocardial infarction (MI) were found for the following parameters: Systolic and diastolic blood pressure, family history of premature MI, cigarette smoking, plasma levels of triglycerides, VLDL-cholesterol and blood glucose. Plasma concentrations of HDL-cholesterol and apo A1 showed an inverse relationship to the MI incidence and--somewhat weaker--the same was true for the frequency of alcohol consumption and of physical leisure activity. Besides LDL-cholesterol, total cholesterol and apoprotein B the following parameters showed significantly positive associations to the risk of chronic coronary heart disease: Systolic as well as diastolic blood pressure and--to a lesser extent--the plasma levels of triglycerides, VLDL-cholesterol and blood glucose. An inverse relationship to the incidence rate of chronic coronary heart disease was found for HDL-cholesterol, apo A1 and the frequency of physical leisure activity. Based on the epidemiological results from GRIPS, additionally considering results from other epidemiologic studies and pathophysiological findings a diagnostic strategy was developed for the early recognition of patients at increased coronary risk. LDL-cholesterol is the most important variable in this diagnostic schedule but the coronary status and the individual profile of further risk factors (i.e. positive family history of premature myocardial infarction, hypertension, diabetes mellitus, cigarette smoking, increased plasma levels of triglyceride rich lipoproteins or Lp(a) as well as decreased plasma concentrations of HDL-cholesterol or apo A1) are additionally taken into account.

In the present study the association between angiographically demonstrated coronary artery disease lipid subfractions and insulin in Caucasians, Asian migrants to the UK and Asians in India was studied. Patients having at least one angiographic lesion of more than 50% were recruited and angiograms scored by two independent observers, blindly. There were 87 Caucasian, 83 British Asian and 30 Indian Asian patients. Lipid subfractions measured were cholesterol, HDL cholesterol, triglyceride, apo A1, apo B, Lp(a). Estimation of glucose and insulin was carried out at fasting and after two hours of oral 75 g glucose. Asians were younger than Caucasians. They did not differ in their body mass index, systolic and diastolic blood pressure or cigarette consumption. In all three ethnic groups cholesterol, LDL cholesterol and apo B showed significant positive association. Insulin levels at fasting or post glucose challenge failed to show any association. Apo B may be an additional marker for coronary artery disease in the ethnic groups studied.

Lipid-lowering effect of Rhus coriaria L. (Rhus) has been investigated in multiple animal studies with promising results. Nonetheless, its clinical efficacy has not been adequately examined.

The aim of this study was to evaluate the lipid-lowering effects of Rhus among patients with hyperlipidemia.

The study was designed as a two-arm, double-blind placebo-controlled randomized clinical trial, using a parallel design. Eighty patients with primary hyperlipidemia were randomly assigned to receive Rhus capsules or placebo for 6 weeks.

The serum lipid levels, apolipoprotein-A1 (Apo-A1) and apolipoprotein-B (Apo-B) were measured.

Mean serum high-density lipoprotein cholesterol (HDL-C) and Apo-A1 levels were significantly increased in the Rhus group, compared with the placebo group, after 6 weeks of intervention (P = 0.001). The analysis of covariance test including age, gender, body mass index (BMI), and smoking as co-variables revealed that the increase in HDL-C and Apo-A1 levels remained significant, and increases in HDL-C were dependent on the increase in Apo-A1 levels. No significant difference was observed between Rhus and placebo groups in terms of mean reductions in total cholesterol, low-density lipoprotein cholesterol and triglyceride levels; however, more significant improvement was observed among obese patients (BMI ≥ 30 kg/m2).

The study showed significant increases in HDL-C and Apo-A1 levels in response to Rhus supplementation in patients with hyperlipidemia.

ClinicalTrials.gov ID: NCT02295293.

BACKGROUND

APOE polymorphism is believed to confer susceptibility to coronary heart disease (CHD) and Alzheimer's disease. It is well known that patterns of APOE polymorphisms differ between populations and ethnic groups, although most of the data available so far have been in whites.

METHODS

We evaluated the frequencies of APOE genotypes and their relationships with serum levels of lipids, lipoproteins and apolipoproteins in two groups of Gulf Arab citizens: a control population of healthy voluntary blood donors (n=106), and a group of patients presenting to the lipid clinic for the first time with combined hyperlipidaemia (CH) (n=41). In both groups, fasting serum total cholesterol (TC), triglycerides (TG), HDL, LDL and apolipoprotein A1 and B levels were measured by routine autoanalyzer methods, and APOE genotyping was performed by validated PCR methods. The lipid and lipoprotein levels were related to the specific APOE allele frequencies.

RESULTS

Allele frequencies were 5.7% for *E2, 85.4% for *E3, and 9.0% for *E4 in the healthy blood donor group. An essentially similar pattern was seen in the patients with CH. This APOE allelic distribution conforms to patterns described in Chinese, whites and South Asians. In both the blood donor and CH groups there were no consistent links between specific APOE pattern and serum lipoproteins, as would have been predicted from APO *E2 and APO *E4 frequencies.

CONCLUSIONS

We conclude that APOE allelic patterns in healthy Kuwaiti blood donors and a smaller group of patients with CH do not satisfactorily predict circulating blood levels of lipids and lipoproteins.

OBJECTIVE

To explore the role of both inheritable and epidemiological factors in the pathogenesis of familial gallstone disease in pedigrees.

METHODS

135 pedigrees, with 695 members aged>> or = 18 (18 - 83, with a mean of 50 +/- 14), 282 males and 413 females, including 370 patients, at least one patient existing in every generation, were investigated. Inquiry was carried out to collect the personal and medical history. Physical examination and ultrasonography were conducted. Blood samples were collected to detect the total cholesterol (TCh), triglyceride (TG), high-density lipoprotein cholesterol (HDL-Ch), and apolipoprotein (Apo) A1 and ApoB. The concentration of low-density lipoprotein cholesterol (LDL-Ch) was calculated by the formula: Tch-TG/5 + HDL-Ch. The inheritable characteristics and heritability were analyzed.

RESULTS

The prevalence of gallstone disease was 53.24%, 56.66% among the females, significantly higher than that among the males (48.23%, P = 0.03). The heritability in the first-degree relatives was 138% +/- 7%, and the concordance rate of monozygotic twins was 100%. The significant risk factors were female gender (P = 0.03), body mass index (P < 0.01) and diet (P < 0.008). The history of hypertension, hyperlipidemia, and diabetes were correlated to gallstone disease (P = 0.008, < 0.008, and 0.03 respectively). Serum HDL-Ch and ApoA1 concentrations were lower in the stone group than in the non-stone group (P = 0.003 and 0.005 respectively). There were no significant differences in TCh, TG, LDL-Ch, and ApoB between the 2 groups.

CONCLUSIONS

Genetic factors play a major role in the pathogenesis of familial gallstone disease, characterized by autosomal dominant inheritance. Female gender, obesity, diet, hypertension, hyperlipidemia, diabetes may be the risk factors of gallstone disease. Dyslipidemia is a characteristic of gallstone disease.

A cohort of 13 female and 14 male heterozygotes for ATP binding cassette A1 (ABCA1) gene defects was directly compared with 13 and 14 unaffected female and male family members of almost exact same age. The activities of three proteins that play key roles in HDL metabolism were measured in addition to extensive lipid and (apo) lipoprotein subfraction analysis. Compared to controls, LCAT activity was reduced by 15% in affected subjects (P < 0.001) while PLTP activity was unaffected. Interestingly, CETP activity was elevated by 50% in the heterozygote siblings of one kindred but was unaffected in heterozygotes of the three other families. With respect to lipids, the heterozygotes had normal total cholesterol (TC), and LDL-cholesterol concentrations but presented with a trend towards increased triglyceride levels (13%; P = 0.08). HDL metabolism, by contrast, was severely affected as illustrated by 40% reductions in HDL-cholesterol (P < 0.001) with concomitant reductions in apoAI (25%; P < 0.001) levels and in lipoprotein subfraction LpAI (28%; P < 0.001), LpAI:AII (24%; P=0.014), and LpCIII:nonB (34%; P < 0.001) concentrations. We furthermore observed reduced average HDL particle size (5%; P = 0.004; 16% in female and 3.6% in male) and reduced plasma apoCIII concentration (15%; P = 0.006) while apoAII, apoAIV, apoE and apoB levels were unchanged. In conclusion, heterozygosity for ABCA1 defects was associated with reduced LCAT activity in absence of effects on PLTP activity. Of special interest was our finding that the effects of compromised ABCA1 function on HDL were more pronounced in women than in men.

A screening study was performed on 106 children with familial risk for coronary heart disease (CHD) and on matched controls. The two groups differed in several parameters. Children of CHD patients exhibited significantly elevated levels of Lp(a) and total cholesterol, reduced HDL apo A1 and apo A2 and increased values of serum hexuronic acid. These results support the concept that genetic and familial factors contribute to the risk of atherosclerosis.

Premenopausal women are known to have less heart disease than their menopausal counterparts and men. However, there is a rising prevalence of coronary artery disease (CAD) in premenopausal females, which necessitates determination of risk factors that negate the effects of hormonal protection. There are few studies describing the prevalence of traditional and emerging risk factors in premenopausal women with CAD. Thus, our objective was to explore the prevalence of traditional and emerging risk factors and features of coronary lesions in premenopausal women with CAD in an Indian population. Forty premenopausal female patients with angiographically proven CAD and undergoing treatment with conventional therapies and 40 age-matched premenopausal females without any evidence of CAD were enrolled. Premenopausal females with CAD most commonly had the single-vessel CAD and the left anterior descending artery was most commonly involved. The prevalence of hypertension, diabetes, obesity, metabolic syndrome, family history of CAD and 10-year risk score was higher in premenopausal females with CAD than controls. Even after treatment with conventional therapies, premenopausal women with CAD had dyslipidemia and significantly elevated levels of emerging risk factors such as ApoB, ApoB/ApoA1 ratio, hsCRP, lipoprotein (a), uric acid, T4, fibrinogen, and total leukocyte count as compared to controls (p < 0.05). Further, they had significantly lower levels of HDL-C, and Apolipoprotein A1 and T3 which are protective markers for vascular risk. Multivariate regression analysis demonstrated that low levels of Apo A1 and high levels of fibrinogen, hsCRP and TG drive the vascular risk, and therefore these factors should be considered as candidates for better diagnosis, early detection, and intervention of CAD in premenopausal women.

The efficacy and safety of simvastatin were evaluated in an open multicenter study over a 24-week period. One hundred seventy-two patients (91 men, 81 women) with primary hypercholesterolemia (mostly polygenic) were enrolled in 14 Centers in Northern Italy. The mean age was 55.8 +/- 9.7 years and the mean baseline total cholesterol level was 305 +/- 59 mg/dL. After 4 weeks on an AHA step 1 diet, patients who met the inclusion criterion (total cholesterol>> or = 250 mg/dL) were given simvastatin 10 or 20 mg in the evening. The dose could be titrated up to a maximum of 40 mg o.d. at week 6 and 12. No dose titration was allowed after week 12. One hundred forty-nine patients (86.6%) completed the study according to the protocol, 2 (1.2%) were withdrawn from the study because of adverse events not related to the drug, 21 (12.2%) were unavailable for follow-up. Simvastatin treatment was associated with a sustained dose-related reduction in total and LDL cholesterol (-28% and -39% respectively at the end of the study). Triglycerides showed a significant descending trend (-16% at week 24) and HDL-C increased by 9%. Apolipoproteins were measured in only 25 patients: apo B was reduced by 30% and apo A1 increased by 9%. Clinical side effects were not relevant. Mean levels of GOT, GPT and CPK significantly increased after 6 weeks on simvastatin, but remained stable and at any rate ioitlin the normal range thereafter. Eight patients (5.4%) experienced small transaminase level elevations (< 3ULN) and six (4%) small CPK elevations.(ABSTRACT TRUNCATED AT 250 WORDS)

A heme-dependent conformational rearrangement of the C-terminal domain of heme binding protein (PhuS) is required for interaction with the iron-regulated heme oxygenase (HemO). Herein, we further investigate the underlying mechanism of this conformational rearrangement and its implications for heme transfer via site-directed mutagenesis, resonance Raman (RR), hydrogen-deuterium exchange MS (HDX-MS) methods, and molecular dynamics (MD). HDX-MS revealed that the apo-PhuS C-terminal α6/α7/α8-helices are largely unstructured, whereas the apo-PhuS H212R variant showed an increase in structure within these regions. The increased rate of heme association with apo-PhuS H212R compared with the WT and lack of a detectable five-coordinate high-spin (5cHS) heme intermediate are consistent with a more folded and less dynamic C-terminal domain. HDX-MS and MD of holo-PhuS indicate an overall reduction in molecular flexibility throughout the protein, with significant structural rearrangement and protection of the heme binding pocket. We observed slow cooperative unfolding/folding events within the C-terminal helices of holo-PhuS and the N-terminal α1/α2-helices that are dampened or eliminated in the holo-PhuS H212R variant. Chemical cross-linking and MALDI-TOF MS mapped these same regions to the PhuS:HemO protein-protein interface. We previously proposed that the protein-protein interaction induces conformational rearrangement, promoting a ligand switch from His-209 to His-212 and triggering heme release to HemO. The reduced conformational freedom of holo-PhuS H212R combined with the increase in entropy and decrease in heme transfer on interaction with HemO further support this model. This study provides significant insight into the role of protein dynamics in heme binding and release in bacterial heme transport proteins.

Acetone precipitation was evaluated as a rapid, simple, low-cost, and efficient method for the selective purification of O-glycopeptides from enzymatic digests of glycoproteins. Ovalbumin (OVA), human and bovine α1-acid glycoprotein (hAGP and bAGP), human apolipoprotein C-III (APO-C3), and recombinant human erythropoietin (rhEPO) were used to obtain enzymatic digests with a broad and varied set of peptides, N-glycopeptides, and O-glycopeptides. After digestion and before capillary electrophoresis mass spectrometry (CE-MS) analysis, the amount of ice-cold acetone added to the digests was optimized to maximize recoveries of O-glycopeptides. Furthermore, the different behavior of peptides, N- and O-glycopeptides was explained by studying with multivariate data analysis methods the influence of several physicochemical parameters and properties related to their composition and structure. Principal component analysis (PCA) and, afterward, partial least-squares discriminant analysis (PLS-DA) were used to identify the most significant variables and their importance to differentiate between peptides, N-glycopeptides and O-glycopeptides, or within these classes. This information was useful to understand precipitation of these compounds after addition of acetone and for the selection of the optimal conditions for purification of specific O-glycopeptide biomarkers. Special attention was paid to O126-glycopeptide glycoforms of rhEPO because of their applicability in biopharmaceutical quality control and doping analysis.

To assess bezafibrate efficacy in a diabetic population a single-blind randomized study was performed in 32 diet-resistant type IIb hyperlipidaemic non-insulin-dependent (NID) diabetic patients in good metabolic control (HbA1c < 8%) compared to a placebo group. In our diabetic patients one month treatment of 400 mg/day bezafibrate lowered plasma C (-14%) and TG (-37%) and globally reduced the VLDL particles and VLDL lipids (-37% for C, -56% for TG and -25% for PL), raising VLDL C/TG ratio (+46%), redistributing TG from VLDL to LDL (+10%) and mainly in HDL (+49%), lowered LDL-C and Apo B levels and increased HDL-C together with Apo A1 (+19% and +13%) and Apo A1/Apo B (+72%). PL were raised by bezafibrate treatment and were redistributed from VLDL (-25%) to LDL (+25%) and HDL (+18%), while PL/C ratio increased in VLDL and in LDL (+18% and +50% respectively). Bezafibrate use was safe and improved the lipid pattern and the apolipoprotein and lipid distribution in the lipoproteins, producing a less atherogenic pattern in our NID diabetics.

Liver cells absorb apolipoprotein (Apo) B48-carrying lipoproteins in ApoE's absence, albeit not as efficiently as the ApoE-mediated process. Our objective was to identify differentially expressed hepatic endosome proteins in mice expressing ApoB48 but lacking ApoE and ApoB100 expression (ApoE-/-/B48/48). We purified early and late endosomes from ApoE-/-/B48/48 and wild-type mouse's livers. In ApoE-/-/B48/48 mouse's hepatic endosomes, proteomic analysis revealed elevated protein levels of major urinary protein 6 (MUP), calreticulin, protein disulfide isomerases (PDI) A1, and A3. These proteins are capable of interacting with lipids/lipoproteins and triggering receptor-mediated endocytosis. In addition, hepatic endosomes from ApoE-/- /B48/48 mice exhibited significantly reduced protein levels of haptoglobin, hemopexin, late endosome/lysosome interacting protein, cell division control protein 2 homolog, γ-soluble Nethylmaleimide- sensitive factor attachment protein, vacuolar ATP synthase catalytic subunit A1, dipeptidyl peptidases II, cathepsin B, D, H, and Z. These proteins participate in plasma heme clearance, receptor-mediated signaling, membrane fusion, endosomal/lysosomal acidification, and protein degradation. The significance of increasing endosomal MUP, calreticulin and PDIs in ApoE-/-/B48/48 mouse liver cells is not clear; however, reducing endosomal/ lysosomal membrane proteins and hydrolases might be, at least partially, responsible for the retarded clearance of plasma ApoB-carrying lipoproteins in ApoE-/-/B48/48 mice.