Familial hypercholesterolemia (FH) is an autosomal codominant disorder characterized by impaired clearance of low-density lipoproteins from the bloodstream, markedly elevated plasma total cholesterol and low density lipoprotein cholesterol levels, and early onset of atherosclerosis and cardiovascular disease. FH is one of the most common monogenic disorders in humans. The majority of FH cases are caused by pathogenic variants in three genes:
LDLR
(OMIM: 143890),
APOB
(OMIM: 107730), and
PCSK9
(OMIM: 607786). More than 80 % of FH cases are associated with mutations in the
LDLR
gene, located on chromosome 19. In 25–75 % of patients with a clinical FH phenotype, no pathogenic variant is identified in these genes. Whole-exome sequencing and targeted gene panel sequencing of the
LDLR
,
APOB
,
PCSK9
, and
LDLRAP1
genes were performed in patients with an FH phenotype, followed by confirmation of the identified variants by Sanger sequencing. Three unrelated probands were found to carry intronic
LDLR
variants for which functional evidence was previously unavailable. The aim of this study was to functionally assess
in vitro
the effects of three intronic
LDLR
variants (NM_000527.5: c.940+3_940+6del, c.941-3C>G, and c.2389+5G>A) on pre-mRNA splicing using a minigene assay. Minigene constructs encompassing target exons with flanking intronic sequences were generated and transfected into the HEK293 and HeLa cell lines. The deleterious effect of all three variants on pre-mRNA splicing was confirmed. The fournucleotide deletion c.940+3_940+6del resulted in two aberrant transcripts: inclusion of six nucleotides from intron 6 and complete retention of the minigene intronic sequence. The c.941-3C>G variant caused loss of the canonical acceptor splice site and activation of a cryptic site, with inclusion of intronic nucleotides from intron 6. The c.2389+5G>A variant resulted in exon 16 skipping. Functional
in vitro
analysis is a key tool for the molecular verification of intronic variants of uncertain clinical significance and, in conjunction with clinical data, supports the establishment of a definitive molecular diagnosis.
V. Danilchenko, D. Ivanoshchuk, O. Timoshchenko et al.· Vavilov Journal of Genetics...· 0 citations
Aim. To evaluate the effectiveness of outpatient monitoring of patients with arterial hypertension using an automatic remote blood pressure (BP) monitoring system and compare it with the effectiveness of traditional self-monitoring of BP in a metropolitan urban clinic.
Design. Prospective, interventional, randomized, open-label study.
Materials and methods. We recruited 60 patients (40% men, aged 25–65 years, median age 59.5 [48.5; 69.0] years) undergoing outpatient monitoring with a diagnosis of stage 1 or 2 essential (primary) hypertension. Participants were randomized into two groups using the envelope randomization method. The main group consisted of 30 people who were given certified automatic tonometers for measuring BP with remote online data transmission to the physician's computer. The control group included 30 patients who measured their blood pressure at home using their own automatic tonometers and completed self-monitoring diaries. Pulse rate was recorded two or three times, simultaneously with blood pressure measurements. All patients underwent a comprehensive examination, including a general clinical and biochemical blood test, and a urinalysis. Adherence to therapy was assessed using the Morisky — Greene scale, and anxiety and depression were assessed using the Hospital Anxiety and Depression Scale. The observation period was 6 months, during which two follow-up visits were conducted: on day 3 and at 180 ± 2 days, and a phone call at 90 ± 2 days.
Results. When assessing the dynamics of office BP, a statistically significant decrease in systolic and diastolic BP was found in the main and in the control groups (for all indicators p < 0.001) at visit 3 compared with the values at visit 1. By visit 3, in the main group, target values of outpatient BP were achieved in 26 (86.7%) patients, while in the control group — in 23 (76.7%) (p = 0.011). In the main group, the number of patients prescribed drugs in the form of fixed-dose combinations increased from 15 (50%) to 22 (73.3%) people (p < 0.001), in the control group — from 15 (50%) to 19 (63.3%) people (p < 0.001). An intergroup comparison revealed a statistically significant difference in the frequency of taking fixed-dose combinations of AHT agents at visit 3: 22 (73.3%) patients in the main group and 19 (63.3%) patients in the control group (p = 0.005). In patients in the main group, the severity of anxiety (p = 0.016) and depression (p = 0.014) decreased significantly from visit 1 to visit 3, while in the control group — only depression (p = 0.036). According to the Morisky — Greene scale, adherence to therapy in the main group increased statistically significantly (p = 0.007). Improved adherence to treatment was noted in 29 (96.7%) patients. In an intergroup comparison at visit 3, the number of points on the Morisky — Greene scale was statistically significantly lower in the main group than in the control group: 1 [0; 1] versus 3 [2; 3] points (p = 0.001).
Conclusion. Remote BP monitoring is a promising method that can significantly improve hypertension control in real-life clinical practice in patients aged 25–65 years.
Keywords: telemedicine, telemonitoring, remote monitoring of blood pressure, dispensary observation, arterial hypertension
O. Tsygankova, O. Timoshchenko, L. D. Latyntseva et al.· Doctor Ru· 0 citations
