Lipid Assays Research Articles

Cost of Integrating Noncommunicable Disease Screening Into Home-Based HIV Testing and Counseling in South Africa.

Integrated HIV-noncommunicable disease (NCD) services have the potential to avert death and disability but require data on program costs to assess the impact of integrated services on affordability. We estimated the incremental costs of NCD screening as part of home-based HIV testing and counseling (HTC) and referral to care in KwaZulu-Natal, South Africa. All adults in the households were offered integrated HIV-NCD screening (for HIV, diabetes, hypertension, hypercholesterolemia, obesity, depression, tobacco, and alcohol use), counseling, and linkage to care. We conducted comprehensive program microcosting including ingredient-based and activity-based costing, staff interviews, and time assessment studies. Sensitivity analyses varied cost inputs and screening efficiency. Integrating all-inclusive NCD screening as part of home-based HTC in a high HIV prevalence setting increased program costs by $3.95 (42%) per person screened (from $9.36 to $13.31 per person). Integrated NCD screening, excluding point-of-care cholesterol testing, increased program costs by $2.24 (24%). Furthermore, NCD screening integrated into HTC services reduced the number of persons tested by 15%-20% per day. Integrated HIV-NCD screening has the potential to efficiently use resources compared with stand-alone services. Although all-inclusive NCD screening could increase the incremental cost per person screened for integrated HIV-NCD services over 40%, a less costly lipid assay or targeted screening would result in a modest increase in costs with the potential to avert NCD death and disability. Our analysis highlights the need for implementation science studies to estimate the cost-effectiveness of integrated HIV-NCD screening and linkage per disability-adjusted life year and death averted.

Discovery and characterization of a novel HCV inhibitor targeting the late stage of HCV life cycle.

Currently approved anti-HCV drugs, the direct-acting antivirals (DAAs), are highly effective and target the viral RNA replication stage of the HCV life cycle. Due to high mutation rate of HCV, drug resistant variants can arise during DAA monotherapy. Thus, a combination of DAAs is necessary to achieve a high response rate. Novel HCV inhibitors targeting the HCV late stage such as assembly and release may further improve combination therapy with the DAAs. Here we characterize one late stage-targeting candidate compound, 6-(4-chloro-3-methylphenoxy)-pyridin-3-amine (MLS000833705). We treated HCV-infected cells with MLS000833705 and other HCV inhibitors and examined HCV RNA and infectious titres. We evaluated the colocalization of HCV core and lipid droplets by confocal microscopy. We performed HCV core-proteinase K digestion assay and several lipid assays to study the mechanism of MLS000833705. We showed that MLS000833705 decreased extracellular HCV RNA levels more than intracellular HCV RNA levels in HCV infectious cell culture. Similarly, MLS000833705 reduced infectious HCV titres substantially more in the culture supernatant than intracellularly. Confocal microscopy showed that MLS000833705 did not affect the colocalization of HCV core protein with cellular lipid droplets where HCV assembles. HCV core-proteinase K digestion assay showed that MLS000833705 inhibited the envelopment of HCV capsid. Our study demonstrates that MLS000833705 is a late-stage HCV inhibitor targeting HCV morphogenesis and maturation. Therefore, MLS000833705 can be used as a molecular probe to study HCV maturation and secretion and possibly guide development of a new class of HCV antivirals.

Effect of intranasal insulin on peripheral glucose profile in dexamethasone-induced insulin resistance in Wistar rats

This study evaluates the therapeutic potential of intranasal insulin (INI) in dexamethasone-induced insulin resistance in Wistar rats. In the first phase of the study, thirteen, healthy, untreated male Wistar rats were divided into 2 groups and administered either vehicle (0.9% Normal saline, 20 µl) or insulin (2 IU) intranasally to assess intranasal delivery of insulin in brain. In the second phase of experiments, to evaluate the acute effects of intranasal insulin on peripheral blood glucose, intranasal or intraperitoneal insulin was co-administered with or without dexamethasone 10 mg/kg to 26 male Wistar rats and blood glucose monitored. To evaluate effect of intranasal insulin in peripheral metabolic disease model, insulin or vehicle was administered via intranasal or intraperitoneal (IP) route to control or dexamethasone (Dex)-treated (0.5 mg/kg, IP) female Wistar rats for seven consecutive days. Twenty-four hours after last dose, trunk blood was collected via cardiac puncture. Biochemical assay of glucose, lipid and insulin was performed on serum while enzyme activity – glucokinase and glucose-6-phosphatase (G6Pase) or lactate dehydrogenase (LDH) and glucose-6-phosphate dehydrogenase (G6PDH) were assayed from liver and brain homogenates respectively. Acute intranasal but not intraperitoneal insulin elevated brain insulin after 30 min. In animals administered single dose of 10 mg/kg dexamethasone, intranasal and intraperitoneal insulin lowered blood glucose within one hour. However, only the former’s effect was maintained at the 3rd and fourth hour. Dex-induced hyperglycemia was associated with increased hepatic glucose-6-phosphatase activity and decreased high-density lipoprotein (HDL), these effect were attenuated by subchronic (INI) administration. Also INI did not induce oxidative stress in the brain which suggests no brain damage during the period of study. Subchronic administration of INI was able to reduce the effect of Dex-induced hyperglycemia that is associated with increased hepatic glucose-6-phosphatase activity and decreased high-density lipoprotein (HDL) without damage to the brain. We demonstrate the potential of brain targeting with intranasal insulin in a rat model of insulin resistance and peripheral metabolic disease.

Abstract 397: Cotinine, but Not Nicotine, Increased ApoA-I Mediated HDL Synthesis in THP-1 Cells

Aim: Cotinine is known as a stable metabolite of nicotine, the plasma half-life is over 20-30 hrs. Here, we evaluated its characteristic properties on the apoA-I mediated HDL generation. Method: Cotinine (5.6 to 11.2μM), or nicotine (5 to 150μM) were supplemented to medium and co-incubated with apoA-I (10μg/mL) for 18 hrs with THP-1 cells to determine their effect on efflux activity. Cellular ACAT accessible cholesterol pool size was determined by [ 14 C]oleic acid labeling of cells for 60 min immediately after removal of the culture medium. The culture medium and the cellular lipids were extracted by organic solvents and cholesterol and phospholipids content were determined. Nicotine was administered orally as an aqueous solution (10 μM) to C57BL/6 mice for 10 days and the plasma lipoprotein profiles were measured by gel-HPLC (tandem TSK-lipopropak XL ) with online enzymatic lipid assay system (Skylight biotech, Inc.). Results: Supplementation of nicotine into the medium showed no effect on the apoA-I mediated cellular cholesterol release from THP-1 cells. On the other hand, cotinine increased apoA-I mediated cellular cholesterol efflux 2.8 fold. ACAT accessible cholesterol pool size was dramatically decreased to 80% and 58% of control with 5 and 10 μM cotinine treatment, respectively, indicating that cotinine effectively increased apoA-I/ABCA1 mediated cellular cholesterol efflux. Mice with oral nicotine administration showed increased plasma HDL by 11.4% (50.1±4.4 and 55.8±3.9 mg/dL for control and nicotine treatment, respectively). Conclusions: Cotinine increased apoA-I mediated cellular cholesterol efflux in THP-1 cells. The cellular ACAT accessible cholesterol pool size reduction is one of the early reactions of apoA-I/ABCA1 mediated cellular cholesterol efflux. Cotinine seemingly modifies cellular factor/factors to induce a change in cholesterol distribution. Mice administrated oral nicotine may convert the compound to cotinine by CYP2A6 and aldehyde oxidase activities in their liver, and which likely increased HDL generation. Further experiments are necessary to examine the direct effect of cotinine on the ABCA1 transporter for increasing HDL assembly system.

Correlation between dyslipidemia and severity of allergic rhinitis

BackgroundAllergic rhinitis is a common problem affecting between 20 and 25% of the population lowering the quality of life (QOL) more than any other disease. Dyslipidemia is known to impact potently the development of atopy as it promotes proatopic Th2 immunity and allergic inflammation.ObjectiveThe aim was to test the correlation between severity of allergic rhinitis and dyslipidemia.Materials and methodsA comparative study carried out on 350 allergic rhinitis patients were subjected to full serum lipid assays, visual analog scale assessing their nasal symptoms, and QOL assessment using a seven-point scale.ResultsPatients were divided into two groups (according to their lipid profile): abnormal dyslipidemia group (33%) and normal lipid profile group (67%).The abnormal dyslipidemia group showed a more intense allergic rhinitis symptoms compared with the normal lipid profile with poor QOL score (1.97).ConclusionDyslipidemia might play an important role in increasing the severity of allergic rhinitis symptoms with impaired patients’ QOL; therefore, its control could achieve better treatment outcomes.

Human ATG3 Binding to Lipid Bilayers. Role of Lipid Geometry and Electric Charge

Specific protein-lipid interactions lead to a gradual recruitment of AuTophaGy-related (ATG) proteins to the nascent membrane during autophagosome (AP) formation. ATG3, a key protein in the movement of LC3 towards the isolation membrane, has been proposed to facilitate LC3/GABARAP lipidation in highly curved membranes. In this work we have performed a biophysical study of human ATG3 interaction with membranes containing phosphatidylethanolamine, phosphatidylcholine and anionic phospholipids. We have found that ATG3 interacts more strongly with negatively-charged phospholipid vesicles or nanotubes than with electrically neutral model membranes, cone-shaped anionic phospholipids (cardiolipin and phosphatidic acid) being particularly active in promoting binding. Moreover, an increase in membrane curvature facilitates ATG3 recruitment to membranes although addition of anionic lipid molecules makes the curvature factor relatively less important. The predicted N-terminus amphipathic α-helix of ATG3 would be responsible for membrane curvature detection, the positive residues Lys 9 and 11 being essential in the recognition of phospholipid negative moieties. We have also observed membrane aggregation induced by ATG3 in vitro, which could point to a more complex function of this protein in AP biogenesis. Moreover, in vitro GABARAP lipidation assays suggest that ATG3-membrane interaction could facilitate the lipidation of ATG8 homologues.

Total membrane lipid assay (MLA): simple and practical quantification of exosomes based on efficient membrane-specific dyes unaffected by proteins

MLA uses two fluorescent dyes for ratiometric determination of exosomes' membrane lipids.

Thyroid Function Test, C- Reactive Protein And Blood Lipids In Subclinical Hypothyroidism Patients Reporting At Surgical Wards

Objective: To investigate the thyroid function, C-reactive protein (CRP) and blood lipid profile in the subclinical hypothyroidism (SH) patients in surgical wards and outpatient department. Study design: Cross sectional study Place and Duration: Department of surgery, Isra University Hospital from August 2014 to May 2017. Materials and Methods: 100 subclinical hypothyroid cases and 100 controls were included according to inclusion and exclusion criteria. Serum TSH level > 6.2 (µIU /ml) with normal free T4 and T3 was considered as subclinical hypothyroidism. Venous blood was taken from ante cubital vein, centrifuged and processed. Sera were used for the assay of thyroid hormones, TSH, blood lipids and C-reactive protein. Data variables were analyzed by Statistix 8.0 (95% confidence interval) (P-value =0.05). Results: Age in controls and cases was noted as 51.3±12.5 and 50.8±11.95 years respectively. SH cases show serum T3, T4 and TSH as 0.89±0.18 µg/dl, 4.96±0.85 µg/dl and 11.95±2.85 µU/ml respectively. Serum total cholesterol, triglycerides and LDLc were raised with concomitant low HDLc in SH cases. C- reactive protein in SH cases was 6.91±3.38 ng/ml compared to controls 2.56±1.51 ng/ml (P=0.0001). Serum TSH showed negative correlation with HDLc, serum T3 and T4. Conclusion: The present study reports dyslipidemia with elevated inflammatory marker of C-reactive protein in subclinical hypothyroid patients.

Human ATG3 binding to lipid bilayers: role of lipid geometry, and electric charge

Specific protein-lipid interactions lead to a gradual recruitment of AuTophaGy-related (ATG) proteins to the nascent membrane during autophagosome (AP) formation. ATG3, a key protein in the movement of LC3 towards the isolation membrane, has been proposed to facilitate LC3/GABARAP lipidation in highly curved membranes. In this work we have performed a biophysical study of human ATG3 interaction with membranes containing phosphatidylethanolamine, phosphatidylcholine and anionic phospholipids. We have found that ATG3 interacts more strongly with negatively-charged phospholipid vesicles or nanotubes than with electrically neutral model membranes, cone-shaped anionic phospholipids (cardiolipin and phosphatidic acid) being particularly active in promoting binding. Moreover, an increase in membrane curvature facilitates ATG3 recruitment to membranes although addition of anionic lipid molecules makes the curvature factor relatively less important. The predicted N-terminus amphipathic α-helix of ATG3 would be responsible for membrane curvature detection, the positive residues Lys 9 and 11 being essential in the recognition of phospholipid negative moieties. We have also observed membrane aggregation induced by ATG3 in vitro, which could point to a more complex function of this protein in AP biogenesis. Moreover, in vitro GABARAP lipidation assays suggest that ATG3-membrane interaction could facilitate the lipidation of ATG8 homologues.

Potential Application of 1H NMR for Routine Serum Lipidome Analysis \u2013Evaluation of Effects of Bariatric Surgery

Routine laboratory lipid assays include simple measurements of total cholesterol, triacylglycerols and HDL. However, lipids are a large group of compounds involved in many metabolic pathways, and their alterations may have serious health consequences. In this study, we used 1H NMR to analyze lipids extracted from sera of 16 obese patients prior to and after bariatric surgeries. We observed a post-surgery decrease in serum concentrations of lipids from various groups. The hereby presented findings imply that 1H NMR is suitable for rapid, simple and non-invasive detection of lipids from 30 structural groups, among them triacylglycerols, phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, total phospholipids, total, free and esterified cholesterol, total and unsaturated fatty acids. NMR-based analysis of serum lipids may contribute to a substantial increase in the number of routinely determined markers from this group; therefore, it may find application in clinical assessment of obese subjects prior to and after bariatric surgeries, as well as in the examination of patients with other metabolic diseases.

Associations of the APOB rs693 and rs17240441 polymorphisms with plasma APOB and lipid levels: a meta-analysis

BackgroundThe associations of the apolipoprotein B gene (APOB) rs693 and rs17240441 polymorphisms with plasma levels of APOB and lipids have been widely explored, but the results were inconclusive. This meta-analysis aimed to clarify the associations of the rs693 and rs17240441 polymorphisms with fasting APOB and lipid levels.MethodsSixty-one studies (50,018 subjects) and 23 studies (8425 subjects) were respectively identified for the rs693 and rs17240441 polymorphisms by searching in PubMed, Google Scholar, Web of Science, Cochrane Library, Wanfang, VIP and CNKI databases. The following information was collected for each study: first author, age, gender, ethnicity, health condition, sample size, genotyping, lipid assay method, mean and standard deviation or standard error of APOB and lipid variables by genotypes. A dominant model was used for this meta-analysis.ResultsThe carriers of the rs693 variant allele (T) had higher levels of APOB [standardized mean difference (SMD) = 0.26, 95% confidence interval (CI) = 0.16–0.36, P < 0.01], triglycerides (TG) (SMD = 0.12, 95% CI = 0.05–0.20, P < 0.01), total cholesterol (TC) (SMD = 0.24, 95% CI = 0.17–0.30, P < 0.01) and low-density lipoprotein cholesterol (LDL-C) (SMD = 0.22, 95% CI = 0.14–0.30, P < 0.01), and lower levels of high-density lipoprotein cholesterol (HDL-C) (SMD = −0.06, 95% CI = −0.11–0.01, P = 0.01) than the non-carriers. The carriers of the rs17240441 deletion allele had higher levels of APOB (SMD = 0.13, 95% CI = 0.06–0.20, P < 0.01), TC (SMD = 0.17, 95% CI = 0.07–0.26, P < 0.01) and LDL-C (SMD = 0.15, 95% CI = 0.07–0.23, P < 0.01) than the non-carriers.ConclusionsThe rs693 polymorphism is significantly associated with higher levels of APOB, TG, TC and LDL-C, and lower levels of HDL-C. The rs17240441 polymorphism is significantly associated with higher levels of APOB, TC and LDL-C. Further studies are needed to elucidate the underlying mechanisms.

AGE AND GENDER PECULIARITIES OF BLOOD LIPIDS IN URBAN RESIDENTS (CROSS-SECTIONAL STUDY RESULTS)

Aim . To analyze the results of measurements of the blood lipid spectrum of urban residents of the European part of the Russian Federation. Material and methods . The results of single assessment of lipid profiles in 22,436 males and 35,100 females at the age from 13 to 94 years in 347 cities and towns of the European part of the Russian Federation were analyzed. Lipid assays were performed by the Laboratory Service «HELIX» from 28 Jan 2015 to 19 Mar 2016. Statistical methods included descriptive statistics, the analysis of distributions of sample comparisons and the search of dependencies. Results. Results of 56,235 measurements of total cholesterol (TC), 22,641 – triglycerides (TG), 21,032 – high density lipoproteins cholesterol (HDL-C) and 22,441 – low density lipoprotein cholesterol (LDL-C) were analyzed. TC above 5.2 mmol/l was identified in 55.78% samples; the proportion of people with LDL-C level above 3.0 mmol/l was 62.95%. The level of TC was the highest at the age of 43 to 62 years in both men and women, while the peak in the increase in TC in men was approximately 10 years earlier, than in women. In patients of older age groups, TC levels decreased and reached minimum values in men and women of senile age and long-livers. Low level of HDL-C was observed in 24.46% of the total number of samples in men and in 17.68% – in women. The monotonous increase in the average levels of HDL-C was revealed in men with aging. In women the HDL-C levels increased sharply from a minimum at the age of 13 years to a maximum at the age of 25 years with a slight monotonous decrease in the following age periods. However, even in the age older than 83 years levels of HDL-C remained high (1.49 mmol/l in average). The level of TG showed significant inter-age fluctuations, especially in men. The levels of TG above 1.7 mmol/l were recorded in 30.07% of all cases. Conclusion . Two-factor analysis of variance for all lipid parameters revealed a high statistical significance of gender and age.

Alcohol Consumption Is Associated with Hypogonadism and Decreased Sexual Function in Ghanaian Diabetics

Introduction: Alcohol usage has largely been seen as a risk factor for the development of sexual dysfunction as well as erectile dysfunction. Others have reported that prolonged alcohol usage and abuse is compatible with normal sexual function in the absence of endocrinological problems as well as hepatic dysfunction. About seventy five (75) percent of alcoholics have various sexual difficulties with improvements in sexual functions occurring after treatment of alcoholism and psychosexual therapy. It is evident from the various reports over the years that mild and occasional alcohol usage is not as much implicated in the causation of SD and its other forms as heavy, addictive or dependent alcohol usage. Alcohol usage has also long been linked to hypogonadism, testicular atrophy as well as leydig cell toxicity. Alcohol induced hypogonadism has been reported to resolve after withdrawal of alcohol use. Since both diabetes and alcohol usage have been strongly associated with both hypogonadism and sexual dysfunction, it is logical to expect that diabetics who frequently consume alcohol will have a worsened hypogonadal state and sexual function. This research therefore seeks to provide evidence of an association between alcohol consumption in diabetics and a worsened sexual dysfunction in comparison to diabetics who did not consume alcohol. Methods: Type II diabetic patients attending the Diabetic Clinic at the Maamobi General Hospital between the periods of January 2010 and March 2011 were consecutively recruited for this study. Diabetics with other known endocrinological diseases and physical disabilities were excluded from the study. Sexual function was assessed using the GRISS-M. Early morning fasting samples were used in lipid and testosterone profile assays. Results: Study participants who consumed alcohol recorded higher levels of triglycerides and LDL-Cholesterol. They also recorded significantly lower levels of bioavailable testosterone. Furthermore they also recorded higher scores for impotence, premature ejaculation, non-sensuality and infrequency but lower scores for avoidance and were about six times more likely to be infrequent in their sexual activity in comparison with those who did not consume alcohol. Conclusion: Alcohol consumption among diabetic males is associated with hypogonadism and has an impact on several domains of male sexual function. Diabetic males should be advised to avoid alcohol abuse in order to facilitate the management of diabetes associated sexual dysfunction.

The link between metabolic features and TSH levels in polycystic ovary syndrome is modulated by the body weight: an euglycaemic-hyperinsulinaemic clamp study.

To evaluate the link among thyroid function, glucose/insulin metabolism and steroid hormones in women with polycystic ovary syndrome (PCOS), and to verify if the body mass index (BMI) might influence the interplay between PCOS features and subclinical hypothyroidism (SCH). Case-control study conducted from January to December 2014. One-hundred fifty-four young women with PCOS, according to Rotterdam criteria, and 88 controls were enrolled in an academic research environment. Anthropometric evaluation, hormonal and lipid assays, oral glucose tolerance test (OGTT) and euglycaemic-hyperinsulinaemic clamp were performed. Hirsutism was assessed with the Ferriman-Gallwey (FG) score. SCH was found in 14% of PCOS subjects and in 1% of controls (P < 0.01). In PCOS women, TSH levels were directly correlated with fasting glycaemia, but not with other hormonal and metabolic parameters. When PCOS patients were classified on the basis of BMI, TSH levels significantly correlated with insulin secretion, insulin resistance, DHEAS and cortisol levels in obese PCOS women. Inverse correlations were found between TSH and both oestradiol and SHBG in the same group. In nonobese PCOS patients, only waist-to-hip ratio values were correlated with TSH. The prevalence of SCH was not different between nonobese and obese PCOS groups (14 and 15% respectively). However, SCH was associated with higher levels of insulin, DHEAS, cortisol and FG score only in the obese subgroup. Our data confirm that the prevalence of SCH is increased in PCOS women. The presence of SCH is associated with endocrine and metabolic imbalances of PCOS, and the excessive body weight seems to promote this interplay.

Combined nitrogen limitation and hydrogen peroxide treatment enhances neutral lipid accumulation in the marine diatom Phaeodactylum tricornutum

Exogenous application of dilute hydrogen peroxide (H2O2) increases neutral lipid production in Phaeodactylum tricornutum. Exposing early stationary phase cultures of P. tricornutum to 0.25–2mM H2O2 increases the amount of neutral lipids per biomass (mg/mg) by >100% at 24h post H2O2 treatment as determined upon lipid extraction and analysis using a neutral lipid assay. H2O2 treatment increased the total levels of neutral lipids harvested up to 50%, from 64mg/L to 96mg/L, demonstrating its possible effectiveness as a pre-harvest strategy to enhance the biofuel feedstock potential of P. tricornutum. The effects of H2O2 on biomass are concentration dependent; increasing concentrations of H2O2 reduce the levels of isolated biomass. Analysis of combined stressors demonstrates that H2O2 treatment exhibits synergistic effects to enhance neutral lipid production under nitrogen-depleted, but not phosphorus-depleted conditions, suggesting that the effects of hydrogen peroxide on lipid production are influenced by environmental nitrogen levels.

Purification of Active Respiratory Supercomplex from Bovine Heart Mitochondria Enables Functional Studies

To understand the roles of mitochondrial respiratory chain supercomplexes, methods for consistently separating and preparing supercomplexes must be established. To this end, we solubilized supercomplexes from bovine heart mitochondria with digitonin and then replaced digitonin with amphipol (A8–35), an amphiphilic polymer. Afterward, supercomplexes were separated from other complexes by sucrose density gradient centrifugation. Twenty-six grams of bovine myocardium yielded 3.2 mg of amphipol-stabilized supercomplex. The purified supercomplexes were analyzed based on their absorption spectra as well as Q10 (ubiquinone with ten isoprene units) and lipid assays. The supercomplex sample did not contain cytochrome c but did contain complexes I, III, and IV at a ratio of 1:2:1, 6 molecules of Q10, and 623 atoms of phosphorus. When cytochrome c was added, the supercomplex exhibited KCN-sensitive NADH oxidation; thus, the purified supercomplex was active. Reduced complex IV absorbs at 444 nm, so we measured the resonance Raman spectrum of the reduced amphipol-solubilized supercomplex and the mixture of amphipol-solubilized complexes I1, III2, and IV1 using an excitation wavelength of 441.6 nm, allowing measurement precision comparable with that obtained for complex IV alone. Use of the purified active sample provides insights into the effects of supercomplex formation.

Prevalence of dyslipemia in Aristide le Dantec biochemistry laboratory in Dakar Senegal

IntroductionL'objectif de cette étude était d'évaluer la prévalence des dyslipidémies chez les patients reçus au laboratoire de Biochimie de l'Hôpital Aristide Le Dantec pour le dosage d'un paramètre lipidique au cours de l'année 2013.MéthodesIl s'agit d'une étude rétrospective portant sur 1356 patients âgés de 10 à 94 ans reçus au laboratoire de Biochimie du CHU Le Dantec de janvier à décembre 2013. Etaient inclus dans l'étude, tous les patients ayant au moins un paramètre du bilan lipidique dont les résultats étaient enregistrés dans le registre du laboratoire. Le cholestérol total, le cholestérol HDL, le cholestérol LDL ainsi que les triglycérides ont été dosés grâce à des méthodes enzymatiques sur un automate de Biochimie de type Cobas Integra 400 (Roche Diagnostics).RésultatsLa prévalence des dyslipidémies dans notre population d'étude est de 39,30%. Les prévalences de l'hypercholestérolémie, l'hypoHDLémie, l'hyperLDLémie, l'hypertriglycéridémie et l'hyperlipidémie mixte étaient respectivement : 30,89% ; 7,30% ; 31,19% ; 0,51% ; 7,22%. Les sujets de 40 à 59 ans semblaient être plus exposés et on note une prédominance féminine en ce qui concerne l'hypercholestérolémie (54,17% vs 45,82%), l'hypoHDLémie (54,54% vs45, 45%), et l'hyperlipidémie mixte (51,08% vs 48,97%). Enfin les dyslipidémies étaient fortement corrélées à l'HTA et l'obésité.ConclusionLa forte prévalence des dyslipidémies retrouvée dans notre étude démontre l'intérêt d'étudier la prévalence des facteurs de risque cardio-vasculaires en particulier les dyslipidémies dans la population sénégalaise.

A Simple, High-Throughput Method for Analysis of Ceramide, Glucosylceramide, and Ceramide Trihexoside in Dried Blood Spots by LC/MS/MS.

A unique monophasic extraction system coupled with LC/MS/MS to reduce matrix effects for sphingolipid analysis was developed. A solvent mixture of methanol, acetonitrile, and water was identified to simultaneously extract multiple sphingolipids with broad polarity range. To reduce matrix effects, the targeted sphingolipids were analyzed by liquid chromatography-tandem mass spectrometry (LC/MS/MS). The extraction solvent was used as an isocratic mobile phase in chromatographic separation to eliminate solvent exchange steps and enable high-throughput multiple lipid assay. The assay is linear for ceramide from 0.6 to 9 μg/mL with bias <15 %. The intra-assay coefficient of variation is less than 10 % for concentrations from 1.2 to 9 μg/mL, and less than 25 % for concentrations below 1.2 μg/mL. For glucosylceramide and ceramide trihexoside the linear range is 0.05-3 μg/mL with biases <10 % and <20 %, respectively. The intra-assay coefficient of variation for these analytes is less than 10 % at concentrations from 0.4 to 3 μg/mL, and less than 25 % for concentrations below 0.4 μg/mL.

Development of a liquid chromatography/mass spectrometry assay for the bacterial transglycosylation reaction through measurement of Lipid II.

Transglycosylation is the second to last step in the production of bacterial peptidoglycan. It is catalyzed by a transglycosylation site in class A penicillin-binding proteins (PBPs) or monofunctional glycosyl transferases. Several potential inhibitors have been suggested and need to be tested for activity. In this article, we describe the development and validation of an LC/MS assay for Lipid II, the substrate for transglycosylation. The developed assay can be used to monitor the transglycosylation activity of Staphylococcus aureus PBP2. There was no need for modification of Lipid II with a fluorescent tag that could alter affinity of inhibitors toward Lipid II. Recombinant PBP2 was produced in Escherichia coli and has been tested for activity. This LC/MS method is suitable for a transglycosylation assay for PBP2 and since it is relatively fast, it can be used to test inhibitors.

Modification of plant bait technique for direct diagnosis for Rhizoctonia rice pathogens from Myanmar paddy field soils

A modified baiting technique was conducted for selective isolation, fungal DNA diagnosis and fungal cell lipid assay derived from Myanmar isolates of Rhizoctonia spp., causal agents of rice sheath diseases by trapping selective plant stem segments. Bait plant materials of rice, mat rush and cotton were successfully used to isolate R. solani AG1-IA, R. oryzae and R. oryzae-sativae. Moreover, the three plant materials were also effectively used to detect genomic DNA derived from all Rhizoctonia spp. obtained from Myanmar. Rice segment was the most successful materials for detection of fungal cell lipids including palmitic, stearic and linoleic acids. The results of this experiment demonstrate that bait plant materials of rice, mat rush and cotton were the best useful tools for not only direct isolation, but also fungal DNA diagnosis and cell lipid assay of Myanmar soil environmental conditions.