Simultaneous quantification by double rocket immunoelectrophoresis of apolipoproteins A-I and B in blood spotted on filter paper.

Abstract We describe double rocket immunoelectrophoresis for simultaneous quantification of apolipoprotein A-I (apo A-I) and B (apo B) in blood on filter paper. The apolipoproteins from blood spots on filter paper were eluted with detergents (sodium dodecyl sulfate and Triton X-100). The eluates were subjected to electrophoresis on agarose gel containing antisera against both apolipoproteins. Within- and between-assay CVs for apo B/A-I ratios were less than 5.5% and 7.2%, respectively. The apo B/A-I ratio was influenced by length and temperature of storage. In results for 121 venous blood samples, the apo B/A-I ratios in dried blood spots correlated well with those in serum (r = 0.92) and correlated somewhat with the ratios for low-density lipoprotein/high-density lipoprotein cholesterol in serum (r = 0.87). Of these specimens, 68 were from patients with known familial hypercholesterolemia, all of whom had an apo B/A-I ratio greater than 0.90. We think this method will be of value for detecting familial hypercholesterolemia and possibly familial hyperapobeta- and hypoalphalipoproteinemia.

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Use of filter paper for sample collection and transport in steroid pharmacology

Clinical Chemistry ◽

10.1093/clinchem/43.8.1408 ◽

1997 ◽

Vol 43 (8) ◽

pp. 1408-1415 ◽

Cited By ~ 14

Author(s):

Christopher J Howe ◽

David J Handelsman

Keyword(s):

Filter Paper ◽

Pharmacological Study ◽

Venous Blood ◽

Field Studies ◽

Dried Blood Spots ◽

Capillary Blood ◽

Sample Collection ◽

Blood Spots ◽

Adverse Environmental Conditions ◽

Good Agreement

Abstract Field studies of androgen pharmacology are complicated by the necessity to collect, process, and store blood samples in a central facility. We have assessed the feasibility of using capillary blood spots collected by fingerprick and dried on filter paper for pharmacokinetics and pharmacodynamic measurements with nandrolone and testosterone RIAs modified for extracts from capillary blood spots. Assays on punched spots of 7.9-mm diameter (14.9 μL of dried blood) permitted accurate quantification of testosterone down to 0.4 nmol/L from a single spot and nandrolone down to 0.9 nmol/L from two spots. Stability of the steroids in dried blood spots to adverse environmental conditions, notably increased temperatures, was investigated both in the laboratory and in field studies of dried spots sent through the postal system. Storage or postal transport under moderate conditions appeared to have no deleterious effects on apparent androgen concentrations. However, under extreme conditions of storage at 50 °C for a week or more, or transport to a very hot tropical location, a rise in the final concentration of nandrolone, and, to a lesser extent, testosterone when corrected for tracer recovery, was noticed. These effects were largely due to apparent susceptibility of tritiated tracer, but not unlabeled androgens, to thermal degradation. In a pilot pharmacological study involving intramuscular injection of 100 mg of nandrolone decanoate in 1 mL of arachis oil, nandrolone concentrations in concurrently collected plasma as well as venous and capillary blood spots showed good agreement. Testosterone concentrations in contemporaneously collected plasma and venous blood spots also showed very good agreement. We propose that these methods may allow patients and experimental subjects to self-collect samples at remote or field locations for convenient mailing to a central laboratory for androgen assay. Applications of this methodology are now under way.

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Therapeutic Options for Homozygous Familial Hypercholesterolemia: The Role of Lomitapide

Current Medicinal Chemistry ◽

10.2174/0929867326666190121120735 ◽

2020 ◽

Vol 27 (23) ◽

pp. 3773-3783

Author(s):

Antonina Giammanco ◽

Angelo B. Cefalù ◽

Davide Noto ◽

Maurizio R. Averna

Keyword(s):

Familial Hypercholesterolemia ◽

Low Density Lipoprotein ◽

Safety Data ◽

Real Life ◽

Density Lipoprotein ◽

Microsomal Triglyceride Transfer Protein ◽

Poor Responders ◽

Homozygous Familial Hypercholesterolemia ◽

Good Tolerability ◽

Apo B

Background: Lomitapide (Juxtapid® in US and Lojuxta® in Europe) is the first developed inhibitor of the Microsomal Triglyceride Transfer Protein (MTP) approved as a novel drug for the management of Homozygous Familial Hypercholesterolemia (HoFH). It acts by binding directly and selectively to MTP thus decreasing the assembly and secretion of the apo-B containing lipoproteins both in the liver and in the intestine. Aims: The present review aims at summarizing the recent knowledge on lomitapide in the management of HoFH. Results: The efficacy and safety of lomitapide have been evaluated in several trials and it has been shown a reduction of the plasma levels of Low-Density Lipoprotein Cholesterol (LDL-C) by an average of more than 50%. Although the most common side effects are gastrointestinal and liver events, lomitapide presents generally with a good tolerability and satisfactory patients compliance. Recently, in Europe, to evaluate the long-term safety and efficacy of lomitapide, the LOWER registry (ClinicalTrials.gov Identifier: NCT02135705) has been established in order to acquire informations on HoFH lomitapidetreated patients from “real life” clinical practice. : Furthermore, the observation that lomitapide decreases triglyceride levels may be considered for patients affected by severe forms of hypertriglyceridemia who undergo recurrent episodes of pancreatitis and are poor responders to conventional treatment. Conclusion: Lomitapide represents an innovative and efficacious drug for the treatment of HoFH. Longterm safety data, treatment of pediatric and pregnant HoFH patients and management of severe hypertriglyceridemia still require further investigations.

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Clinical implications and outcomes of the ORION Phase III trials

Future Cardiology ◽

10.2217/fca-2020-0150 ◽

2020 ◽

Author(s):

Julia Brandts ◽

Kausik K Ray

Keyword(s):

Familial Hypercholesterolemia ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Phase Iii ◽

Atherosclerotic Cardiovascular Disease ◽

Low Density Lipoprotein Cholesterol ◽

Clinical Safety ◽

Phase Iii Trials ◽

Phase Ii And Iii ◽

Ongoing Phase

Inclisiran is a siRNA inhibiting hepatic PCSK9 synthesis. As a first-in-class therapy, inclisiran has been assessed within the ORION trial program for its low-density lipoprotein cholesterol (LDL-C) lowering efficacy and clinical safety. Phase II and III trials have shown that inclisiran lowers LDL-C by about 50% with an infrequent dosing schedule in patients with established atherosclerotic cardiovascular disease and those at high risk, including patients with heterozygous familial hypercholesterolemia. Ongoing Phase III trials will provide evidence on longer-term safety and effectiveness, and inclisiran’s efficacy in patients with homozygous familial hypercholesterolemia. Furthermore, the ORION-4 trial will assess inclisiran’s impact on cardiovascular outcomes.

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Homozygous familial hypercholesterolemia in a young woman with dual gene mutations of low-density lipoprotein receptor and proprotein convertase subtilisin/kexin type 9

Journal of Clinical Lipidology ◽

10.1016/j.jacl.2020.01.009 ◽

2020 ◽

Vol 14 (2) ◽

pp. 192-196

Author(s):

Patrizia Suppressa ◽

Concetta Carbonara ◽

Natasha Scialpi ◽

Alessandro Ciavarella ◽

Carlo Sabbà

Keyword(s):

Familial Hypercholesterolemia ◽

Young Woman ◽

Low Density Lipoprotein ◽

Gene Mutations ◽

Density Lipoprotein ◽

Low Density ◽

Lipoprotein Receptor ◽

Homozygous Familial Hypercholesterolemia ◽

Proprotein Convertase ◽

Density Lipoprotein Receptor

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Immunoblot analysis of low density lipoprotein receptors in fibroblasts from subjects with familial hypercholesterolemia.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)33635-4 ◽

1982 ◽

Vol 257 (21) ◽

pp. 13150-13156 ◽

Cited By ~ 29

Author(s):

U Beisiegel ◽

W J Schneider ◽

M S Brown ◽

J L Goldstein

Keyword(s):

Familial Hypercholesterolemia ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Immunoblot Analysis ◽

Low Density ◽

Lipoprotein Receptors

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Decreased production of low density lipoprotein by atorvastatin after apheresis in homozygous familial hypercholesterolemia

The Journal of Lipid Research ◽

10.1016/s0022-2275(20)37137-6 ◽

1997 ◽

Vol 38 (10) ◽

pp. 2071-2078

Author(s):

A D Marais ◽

R P Naoumova ◽

J C Firth ◽

C Penny ◽

C K Neuwirth ◽

...

Keyword(s):

Familial Hypercholesterolemia ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Low Density ◽

Homozygous Familial Hypercholesterolemia

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The LDLR, APOB, and PCSK9 Variants of Index Patients with Familial Hypercholesterolemia in Russia

Genes ◽

10.3390/genes12010066 ◽

2021 ◽

Vol 12 (1) ◽

pp. 66

Author(s):

Alexey Meshkov ◽

Alexandra Ershova ◽

Anna Kiseleva ◽

Evgenia Zotova ◽

Evgeniia Sotnikova ◽

...

Keyword(s):

Familial Hypercholesterolemia ◽

Low Density Lipoprotein ◽

Significant Proportion ◽

Density Lipoprotein ◽

Genetic Diagnostics ◽

Atherosclerotic Cardiovascular Disease ◽

Gene Variants ◽

Systematic Analysis ◽

Pcsk9 Gene ◽

Cholesterol Levels

Familial hypercholesterolemia (FH) is a common autosomal codominant disorder, characterized by elevated low-density lipoprotein cholesterol levels causing premature atherosclerotic cardiovascular disease. About 2900 variants of LDLR, APOB, and PCSK9 genes potentially associated with FH have been described earlier. Nevertheless, the genetics of FH in a Russian population is poorly understood. The aim of this study is to present data on the spectrum of LDLR, APOB, and PCSK9 gene variants in a cohort of 595 index Russian patients with FH, as well as an additional systematic analysis of the literature for the period of 1995–2020 on LDLR, APOB and PCSK9 gene variants described in Russian patients with FH. We used targeted and whole genome sequencing to search for variants. Accordingly, when combining our novel data and the data of a systematic literature review, we described 224 variants: 187 variants in LDLR, 14 variants in APOB, and 23 variants in PCSK9. A significant proportion of variants, 81 of 224 (36.1%), were not described earlier in FH patients in other populations and may be specific for Russia. Thus, this study significantly supplements knowledge about the spectrum of variants causing FH in Russia and may contribute to a wider implementation of genetic diagnostics in FH patients in Russia.

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Improving Familial Hypercholesterolemia Index Case Detection: Sequential Active Screening from Centralized Analytical Data

Journal of Clinical Medicine ◽

10.3390/jcm10040749 ◽

2021 ◽

Vol 10 (4) ◽

pp. 749

Author(s):

Fernando Sabatel-Pérez ◽

Joaquín Sánchez-Prieto ◽

Víctor Manuel Becerra-Muñoz ◽

Juan Horacio Alonso-Briales ◽

Pedro Mata ◽

...

Keyword(s):

Familial Hypercholesterolemia ◽

Diagnostic Yield ◽

Low Density Lipoprotein ◽

Analytical Data ◽

Genetic Diagnosis ◽

Density Lipoprotein ◽

Screening Strategy ◽

Cascade Screening ◽

Active Screening ◽

Lipid Clinic

The majority of familial hypercholesterolemia index cases (FH-IC) remain underdiagnosed and undertreated because there are no well-defined strategies for the universal detection of FH. The aim of this study was to evaluate the diagnostic yield of an active screening for FH-IC based on centralized analytical data. From 2016 to 2019, a clinical screening of FH was performed on 469 subjects with severe hypercholesterolemia (low-density lipoprotein cholesterol ≥220 mg/dL), applying the Dutch Lipid Clinic Network (DLCN) criteria. All patients with a DLCN ≥ 6 were genetically tested, as were 10 patients with a DLCN of 3–5 points to compare the diagnostic yield between the two groups. FH was genetically confirmed in 57 of the 84 patients with DLCN ≥ 6, with a genetic diagnosis rate of 67.9% and an overall prevalence of 12.2% (95% confidence interval: 9.3% to 15.5%). Before inclusion in the study, only 36.8% (n = 21) of the patients with the FH mutation had been clinically diagnosed with FH; after genetic screening, FH detection increased 2.3-fold (p < 0.001). The sequential, active screening strategy for FH-IC increases the diagnostic yield for FH with a rational use of the available resources, which may facilitate the implementation of FH universal and family-based cascade screening strategies.

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