p.(Asp47Asn) and p.(Thr62Met): non deleterious LDL receptor missense variants functionally characterized in vitro.

Familial Hypercholesterolemia (FH) is a common genetic disorder caused most often by mutations in the Low Density Lipoprotein Receptor gene (LDLr) leading to high blood cholesterol levels, and ultimately to development of premature coronary heart disease. Genetic analysis and subsequent cascade screening in relatives allow diagnosis of FH at early stage, especially relevant to diagnose children. So far, more than 2300 LDLr variants have been described but only a minority of them have been functionally analysed to evaluate their pathogenicity in FH. Thus, identifying pathogenic mutations in LDLr is a long-standing challenge in the field. In this study, we investigated in vitro the activity p.(Asp47Asn) and p.(Thr62Met) LDLr variants, both in the LR1 region. We used CHO-ldlA7 transfected cells with plasmids carrying p.(Asp47Asn) or p.(Thr62Met) LDLr variants to analyse LDLr expression by FACS and immunoblotting, LDL binding and uptake was determined by FACS and analysis of mutation effects was assessed in silico. The in vitro activity assessment of p.(Asp47Asn) and p.(Thr62Met) LDLr variants shows a fully functional LDL binding and uptake activities. Therefore indicating that the three of them are non-pathogenic LDLr variants. These findings also emphasize the importance of in vitro functional LDLr activity studies to optimize the genetic diagnosis of FH avoiding the report of non-pathogenic variants and possible misdiagnose in relatives if cascade screening is carried out.

Replacement of cysteine at position 46 in the first cysteine-rich repeat of the LDL receptor impairs apolipoprotein recognition.

BACKGROUND AND AIMS: Pathogenic mutations in the Low Density Lipoprotein Receptor gene (LDLR) cause Familial Hypercholesterolemia (FH), one of the most common genetic disorders with a prevalence as high as 1 in 200 in some populations. FH is an autosomal dominant disorder of lipoprotein metabolism characterized by high blood cholesterol levels, deposits of cholesterol in peripheral tissues such as tendon xanthomas and accelerated atherosclerosis. To date, 2500 LDLR variants have been identified in the LDLR gene; however, only a minority of them have been experimentally characterized and proven to be pathogenic. Here we investigated the role of Cys46 located in the first repeat of the LDL receptor binding domain in recognition of apolipoproteins. METHODS: Activity of the p.(Cys46Gly) LDLR variant was assessed by immunoblotting and flow cytometry in CHO-ldlA7 expressing the receptor variant. Affinity of p.(Cys46Gly) for LDL and VLDL was determined by solid-phase immunoassays and in silico analysis was used to predict mutation effects. RESULTS AND CONCLUSION: Functional characterization of p.(Cys46Gly) LDLR variant showed impaired LDL and VLDL binding and uptake activity. Consistent with this, solid-phase immunoassays showed the p.(Cys46Gly) LDLR variant has decreased binding affinity for apolipoproteins. These results indicate the important role of Cys46 in LDL receptor activity and highlight the role of LR1 in LDLr activity modulation. This study reinforces the significance of in vitro functional characterization of LDL receptor activity in developing an accurate approach to FH genetic diagnosis. This is of particular importance because it enables clinicians to tailor personalized treatments for patients' mutation profile.

Effects of a brief, comprehensive, yoga-based program on quality of life and biometric measures in an employee population: a pilot study.

OBJECTIVE: The aim of this study was to determine whether a comprehensive, yoga-based wellness program could positively affect multiple markers of health and wellness in an employee population. DESIGN: Self-selected employees who enrolled in a new wellness class were invited to participate in a yoga-based wellness program. Participants met six days per week (Monday through Saturday) at 5:10 am. Sessions lasted for at least one hour, and the program was six weeks long. Each session consisted of power yoga interwoven with philosophical concepts and instruction about the benefits of mindfulness, breath, and meditation. Certain classes each week incorporated large and small group sharing, journal writing, and mindful eating exercises. Main outcome measures were biometric measures (height, weight, blood pressure, flexibility, body fat) and quality-of-life measures (physical, emotional, and spiritual well-being). RESULTS: Fifty-nine employees were invited to join the program; 50 consented to participate, of which 37 (74%) attended more than 90% of classes. Participant age ranged from 24 to 76 years. Statistically significant improvements were observed in weight (-4.84 ± 5.24 kg; P < .001), diastolic blood pressure (-2.66 ± 8.31 mm/Hg; P = .03), flexibility score (relative change 11% ± 20.92; P <.001), body fat percentage (-1.94 ± 2.68; P < .001), and overall quality of life (linear analog self-assessment [LASA] score 3.73 ± 8.11; P = .03). CONCLUSIONS: This pilot study suggests that a yoga-based, comprehensive wellness program is both feasible and efficacious in creating positive, short-term improvements in multiple domains of health and wellness for a population of employees.