Founder effects facilitate the use of a genotyping-based approach to molecular diagnosis in Swedish patients with familial hypercholesterolaemia.

AIM: To investigate whether genotyping could be used as a cost-effective screening step, preceding next-generation sequencing (NGS), in molecular diagnosis of familial hypercholesterolaemia (FH) in Swedish patients. METHODS AND RESULTS: Three hundred patients of Swedish origin with clinical suspicion of heterozygous FH were analysed using a specific array genotyping panel embedding 112 FH-causing mutations in the LDLR, APOB and PCSK9 genes. The mutations had been selected from previous reports on FH patients in Scandinavia and Finland. Mutation-negative cases were further analysed by NGS. In 181 patients with probable or definite FH using the Dutch lipid clinics network (DLCN) criteria (score ≥ 6), a causative mutation was identified in 116 (64%). Of these, 94 (81%) were detected by genotyping. Ten mutations accounted for more than 50% of the positive cases, with APOB c.10580G>A being the most common. Mutations in LDLR predominated, with (c.2311+1_2312-1)(2514)del (FH Helsinki) and c.259T>G having the highest frequency. Two novel LDLR mutations were identified. In patients with DLCN score < 6, mutation detection rate was significantly higher at younger age. CONCLUSION: A limited number of mutations explain a major fraction of FH cases in Sweden. Combination of selective genotyping and NGS facilitates the clinical challenge of cost-effective genetic screening in suspected FH. The frequency of APOB c.10580G>A was higher than previously reported in Sweden. The lack of demonstrable mutations in the LDLR, APOB and PCSK9 genes in ~1/3 of patients with probable FH strongly suggests that additional genetic mechanisms are to be found in phenotypic FH.

Plasma tumour necrosis factor-alpha and early carotid atherosclerosis in healthy middle-aged men.

AIMS: Tumour necrosis factor-alpha (TNF-alpha) is a proinflammatory cytokine, which is implicated in some metabolic disorders and may play a role in the development of cardiovascular disease. We examined whether plasma TNF-alpha is related to established cardiovascular risk indicators, plasma levels of soluble cellular adhesion molecules and carotid artery intima-media thickness determined by ultrasound examination in a population-based cohort of 96 healthy 50-year-old men. METHODS AND RESULTS: TNF-alpha and cellular adhesion molecules were measured with enzyme-linked immunosorbent assays. Plasma TNF-alpha concentration was associated with systolic and diastolic blood pressure, degrees of alimentary lipaemia, plasma very low density lipoprotein triglyceride, low density lipoprotein (LDL) cholesterol concentrations and peak LDL particle size. Two indices of insulin resistance as well as all soluble cellular adhesion molecules correlated positively with TNF-alpha. The plasma TNF-alpha concentration was associated with common carotid intima-media thickness in univariate analysis. In contrast, soluble E-selectin and postprandial triglycerides, but not TNF-alpha, were independent determinants of common carotid intima--media thickness. CONCLUSION: The plasma TNF-alpha concentration is associated with degrees of early atherosclerosis and correlates with metabolic and cellular perturbations that are considered important for the vascular process.

A common functional polymorphism (C-->A substitution at position -863) in the promoter region of the tumour necrosis factor-alpha (TNF-alpha) gene associated with reduced circulating levels of TNF-alpha.

Tumour necrosis factor-alpha (TNF-alpha) plays a key role in orchestrating the complex events involved in inflammation and immunity. Accordingly, TNF-alpha has been implicated in a wide range of autoimmune and infectious diseases, but also in conditions such as obesity and insulin resistance. The regulation of TNF-alpha expression in man is indicated to be partly genetically determined. We therefore screened a 1263 bp section of the proximal promoter of the TNF-alpha gene for common genetic variants affecting the transcriptional activity of the gene. Here we report the characterization of a common functional polymorphism in the promoter region of the TNF-alpha gene, a C-->A substitution at position -863. Electromobility shift assays provided evidence for a distinct difference in the binding of monocytic and hepatic nuclear factors to the -863C and -863A alleles. The rare -863A allele was associated with 31% lower transcriptional activity ( P < 0.001) in chloramphenicol acetyltransferase (CAT) reporter gene studies in human hepatoblastoma (HepG2) cells, indicating that the-863C/A polymorphism influences the basal rate of transcription of the TNF-alpha gene in vitro. Allele frequencies were 0.83/0.17 amongst 254 apparently healthy men of Swedish origin, aged 35-50 years. In 156 men, the -863C/A polymorphism was associated with the serum TNF-alpha concentration, carriers of the rare A allele having a significantly lower TNF-alpha level ( P < 0.05). It is concluded that the common-863C/A polymorphism in the promoter region of the TNF-alpha gene is functional in vitro in monocytic and hepatic cells and influences the serum TNF-alpha concentration in vivo in healthy middle-aged men.

The influence of cleft length and palatoplasty on the dental arch and the deciduous occlusion in cases of clefts of the secondary palate.

In patients with clefts of the secondary palate, the effect of surgical reconstruction was studied from the time of the palate repair, at the age of 18-24 months, to the age of 5 years. The material comprised 99 children (37 boys and 62 girls). The material was grouped according to the antero-posterior size of the cleft. The changes of the maxillary dimensions were studied on the casts by linear measurements. The occlusion was analysed by descriptive and numerical methods. In the deciduous dentition the smallest intercanine and intermolar dimensions were recorded in patients with large palatal clefts. A the age of 5, the frequency of anterior crossbite was 38% in patients with large palatal clefts, compared with 19--21% in patients with smaller clefts. In boys with cleft palates, the frequency of anterior crossbite was 13% higher than in girls, in spite of the fact that in the material the incidence of large palatal clefts was lower in boys than in girls. In cases of large clefts of the secondary palate, the incidence of anterior crossbite was 12.5 times higher than in a material of non-cleft patients of the same age.

The influence of primary periosteoplasty on maxillary growth and deciduous occlusion in cases of complete unilateral cleft lip and palate. A longitudinal study from infancy to the age of 5.

The influence of infant periosteoplasty upon the growth of the maxilla, its form and size, and the prevalence of malocclusion in the deciduous dentition was investigated. The material consisted of 66 patients with total unilateral clefts of the primary and secondary palate. Thirty-six had periosteoplasty performed in conjunction with cleft-lip and/or palate repair. Thirty patients were operated upon without periosteoplasty and served as controls. Repair of the lip had a notable effect upon the width of the alveolar cleft and palatal cleft, both in the periosteoplasty cases and in the controls, with no certain difference between the groups. Following lip repair, the anterior width of the alveolar arch was slightly reduced. After palatal repair a further reduction was noted in the deciduous dentition, both in the cases treated with periosteoplasty and in the controls, while the posterior width of the palate across the tuberosities increased during growth. In the deciduous dentition, no differences were found in intercanine and intermolar dimensions between the periosteoplasty cases and the controls. Thus, the new bone formed in the cleft area after periosteoplasty does not seem to withstand the contracting forces introduced by palate surgery. An increased length of the buccal alveolar arch on the cleft side, compared with that on the non-cleft side, was found at both the lip repair and the palate repair in the periosteoplasty cases, as well as in the controls. In the deciduous dentition, this difference was negligible. In the deciduous dentition an anterior position of the lateral maxillary segment proved more common in the periosteoplasty cases than in the controls. On the non-cleft side, there was an increased frequency of mesial occlusion and a corresponding decrease of neutral and distal occlusion in the periosteoplasty cases. No increased frequency of anterior crossbite was found even after repeated periosteoplasty, nor was the maxillary dental-arch length unfavourably influenced. Descriptive analysis of occlusion revealed an increase of buccal crossbite in the periosteoplasty cases of a select group of the widest clefts, treated by repeated periosteoplasty. These cases also had the highest total occlusal score according to the numerical classification, while the total occlusal score after one periosteoplasty in patients with less wider clefts was smaller than in the controls. In all patients who had undergone periosteoplasty new bone formed within the alveolar cleft. A good amount of new bone developed in about half the number of cases.(ABSTRACT TRUNCATED AT 400 WORDS)

Reconstruction of articular cartilage using autologous perichondrial grafts. A preliminary report.

In a pilot study of grown-up rabbits perichondrium from the ear was grafted to the joint surface of cavitas glenoidalis from which the normal articular cartilage had been resected. in all cases regeneration of new cartilage occurred. Five clinical cases of arthritis are reported in which, following removal of the degenerated cartilage and grafting of rib perichondrium, articular cartilage regeneration took place

The pathogenesis of cauliflower ear. An experimental study in rabbits.

Appreciating an imcomplete understanding of the pathogenesis of cauliflower ear, an experimental study was designed to demonstrate the pathophysiology of this deformity. The investigation was conducted in 2-month-old rabbits. In one ear a collection of blood was placed under the raised perichondrium which was then sutured back in place and the skin closed. In the other ear an equal amount of blood was deposited between the intact perichondrium and skin. In the first study new cartilage developed under the perichondrium, but in the ear in which the blood was left above the surface of the perichondrium-covered cartilage, complete resorption of the clot occurred. The cauliflower ear was thus shown to be generating cartilage, arising from a layer of raised perichondrium which was further stimulated by a sero-sanguinous medium. The subperichondrial hematoma was extensively invaded by chondroblasts within 2 weeks, and over a period of 4 weeks the new tissue gradually changed into more mature cartilage. It was a consistent finding that the separated perichondrium retracted, thus causing the original cartilage to rise and buckle over the hamatoma, similar to the picture observed in the human pathology.