Single step PCR for the identification of low density lipoprotein receptor [LDL-R] gene mutations

This study was conducted to determine the common mutation of low density lipoprotein receptor in patients with familial hypercholesterolemia [FH] in our population and identify the different point mutation in the LDL-receptor gene. The main aim of this study was to reduce the cost of PCR without extracting DNA and do the diagnosis at single step. This study was carried out in the period of one year, from 2009- 2011. All the patients selected for this study were from Dr. Ziauddin Hospital, National Institute of Cardiovascular Diseases, and Dr. Rubina Ghani's Pathological and Molecular Laboratories. While collecting the blood sample, the patients were in overnight fasting condition. The clinical and biochemical analysis was performed on hyperlipidemic patients [n=120] to determine the frequency of familial hypercholesterolemia in our population. After lipid profile the patients were selected and direct multiplex PCR [Polymerase chain reaction] was performed from whole blood collected in a single tube using forward and reverse primers of exons 3, 4, 9 and 14 of without extracting DNA. Genomic DNA was extracted from blood samples as well as direct whole ETDA blood of healthy control group and hypercholesterolemia patients to detect mutations in exons 3, 4, 9, and 14 of the LDLR gene, with modification in the technique by using type-specific primers. These results for exon 4 mutation were confirmed by DNA sequencing. Screening method based on PCR by using Kappa direct PCR could be a faster and cheaper method with least contamination for screening a large number of FH patients for mutation of LDLR gene

Determination of site-specific carotid-intima media thickness: common -carotid artery and carotid bifurcation in hypercholesterolemia patients

Objective: To determine site specific carotid intima-media thickness: common-carotid artery and carotid bifurcation in hypercholesterolemia patients as a marker for atherosclerosis

Methods: Fifty patients with hypercholesterolemia and twenty controls were selected after getting informed consent regarding the investigation of carotid- intima media thickness by B-mode ultrasound. All the patients of hypercholesterolemia with LDL-C > 160mg/dL had family history of coronary artery diseases. This procedure was carried out in the Radiology Department of Dr. Ziauddin Hospitals. Measurement of carotid -intima media thickness, B-mode ultrasonography of common carotid artery, carotid bifurcation and internal carotid artery [left and right carotid arteries] was performed with Toshiba [M# SSA-580A/E2] ultrasound scanner with linear probe. The posterior or far wall of the carotid artery is, the distance between the leading edge first bright line [lumen -intima interface] and the leading edge of the second bright line [media-adventitia interface] of far wall was recorded as intima -media thickness. The average mean of six segments of intima-media thickness was taken as mean CIMT of right and left common carotid, bifurcation and internal carotid arteries

Results: Maximal CIMT was significantly increased at sites common carotid, carotid bifurcation and internal carotid arteries in fifty patients with hypercholesterolemia as compared to controls. CIMT comparison of hypercholesterolemia patients and controls was assessed by applying students t-test [p < 0.001***]. At carotid bifurcation mean of maximal CIMT was [0.9+/-0.3mm]. Range of maximum CIMT in hypercholesterolemia patients was [0.8- 3.3mm] and in controls [0.4- 0.8 mm]. The thickness was more frequently increased at site of bifurcation

Conclusions: Carotid intima- media thickness in hypercholesterolemia patients was increased and carotid bifurcation was site that has shown greater increase in intima-media thickness and plaques in these patients predict high risk for atherosclerosis

Carotid intima-media thickness correlation with lipid profile in patients with familial hypercholesterolemia versus controls

To determine the variations in carotid intima-media thickness [CIMT] in familial hypercholesterolemia [FH] patients and its use as predictive marker for premature cardiovascular diseases. National Institute of Cardiovascular Diseases and Dr. Ziauddin Hospital, Karachi, from June 2008 to October 2009. Familial hypercholesterolemia was clinically diagnosed by premature coronary diseases, xanthomas, arcus cornealis and family history of premature coronary heart diseases. Controls were age matched normal individuals without hypercholesterolemia. Their lipid profile was tested after overnight fasting. CIMT was measured in mm using B-mode ultrasonography using linear probe. Student t-test was applied to compare mean CIMT of cases and the control. The mean CIMT values of the FH cases were correlated with LDL using Pearson's correlation test. Forty cases with hypercholesterolemia gave consent to participate in the study. These patients had total cholesterol >200 mg/dL and LDL >/= 160 mg/dL as compared to twenty controls of similar age with total cholesterol

Moxonidine-in mild to moderate systemic hypertension

We have recently concluded a nation wide post marketing study on efficacy and safety of Moxonidine in mild to moderate hypertensives. In all 97 patients were enrolled and out of these 87 completed a 12 weeks trial. There was an average drop of 12.9 mmHg in the DBP and 16.5 mmHg in SBP. 80% patient required a daily dose of 0.2 - 0.4 mg of Moxonidine for control of their BP In 84% of the patients completing the trial the BP changed from moderate to mild or normal or from mild toonormal. The drug was well tolerated, only 2.3% having adverse effects. We conclude that the drug studied is an effective alternate for the control of hypertension