Screening by high-throughput sequencing for pathogenic variants in cystic fibrosis: Benefit of introducing personalized therapies.

This short report documented cystic fibrosis transmembrane conductance regulator (CFTR) variants in 37 patients with cystic fibrosis (CF) in the Rio Grande do Norte region of Northeast Brazil. The high-throughput sequencing technology (HTS) genetic testing provided a definitive molecular diagnosis in 31 patients (83.8%). Among them, 25 patients' carriers of the c.1521_1523delCTT variant, categorized as a class 2 mutation, can be currently treated with CFTR modulator drugs. Five children aged 2-5 years could benefit from double lumacaftor/ivacaftor therapy, and 20 patients aged >6 years could be treated with the triple-combination elexacaftor/tezacaftor/ivacaftor therapy. Thus, the identification of pathogenic variants associated with the development of this disease allows for the introduction of therapy with CFTR modulators that favour better patient management.

Application of high-resolution array platform for genome-wide copy number variation analysis in patients with nonsyndromic cleft lip and palate.

BACKGROUND: Although more than 14 loci may be involved in the development of nonsyndromic cleft lip and palate (NSCLP), the etiology has not been fully elucidated due to genetic and environmental risk factor interactions. Despite advances in identifying genes associated with the NSCLP development using traditional genetic mapping strategies of candidate genes, genome-wide studies, and epidemiologic and linkage analysis, microarray techniques have become important complementary tools in the search for potential causative oral clefts genes in genetic studies. Microarray hybridization enables scanning of the whole genome and detecting copy number variants (CNVs). Although common benign CNVs are often smaller, with sizes smaller than 20 kb, here we reveal small exonic CNVs based on the importance of the encompassed genes in cleft lip and palate phenotype. METHODS: Microarray hybridization analysis was performed in 15 individuals with NSCLP. RESULTS: We identified 11 exonic CNVs affecting at least one exon of the candidate genes. Thirteen candidate genes (COL11A1-1p21; IRF6-1q32.3; MSX1-4p16.2; TERT-5p15.33; MIR4457-5p15.33; CLPTM1L-5p15.33; ESR1-6q25.1; GLI3-7p13; FGFR-8p11.23; TBX1-22q11.21; OFD-Xp22; PHF8-Xp11.22; and FLNA-Xq28) overlapped with the CNVs identified. CONCLUSIONS: Considering the importance to NSCLP, the microdeletions that encompass MSX1, microduplications over TERT, MIR4457, CLPTM1L, and microduplication of PHF8 have been identified as small CNVs related to sequence variants associated with oral clefts susceptibility. Our findings represent a preliminary study on the clinical significance of small CNVs and their relationship with genes implicated in NSCLP.

Gait analysis in short-term follow-up of medial opening wedge high tibial osteotomy.

BACKGROUND: Biomechanical gait changes are proposed as adaptations to medial knee osteoarthritis (OA), and little is known about which parameters can be modified early by high tibial osteotomy (HTO) surgery. The aim of this study was to identify early gait changes in a postoperative period of 6 months as compared to a control group, in three different spatial planes. METHODS: Twenty-one patients with OA were submitted to three-dimensional gait analysis preoperatively and 6 months after HTO surgery. Sixteen healthy individuals were selected for the control group. RESULTS: Compared to the control group, OA patients walked more slowly, with a shorter stride length, and with a higher knee varus and flexion angles. The gait changes detected in the postoperative of 6 months were a significant reduction in knee varus angle and adductor moment in coronal plane; an important reduction in knee extension and an increased extensor moment in sagittal plane; also an increased foot external rotation angle in axial plane was observed. Flexion angle peak in swing phase, adductor and flexor moments were the gait parameters with postoperatively results that were closer to those of the control group. CONCLUSIONS: Even in a short follow-up of 6 months, HTO determines positive results in biomechanical gait, not only in the coronal plane but also in the sagittal and axial planes and should be taken into account during the rehabilitation process.

Low bone mineral density in patients with type 1 diabetes: association with reduced expression of IGF1, IGF1R and TGF B 1 in peripheral blood mononuclear cells.

BACKGROUND: The negative effects of type 1 diabetes (T1D) on growth factors of bone metabolism lead to a reduction in bone mineral density. This study aimed to evaluate the association between bone mineral density and insulin-like growth factor 1 (IGF1), insulin-like growth factor 1 receptor (IGF1R) and transforming growth factor beta 1 (TGFB1) expressions in children and adolescents with T1D. Moreover, the influences of age at diagnosis, time since diagnosis, glycaemic control and albuminuria on bone mineral density were investigated. METHODS: Eighty-six T1D children/adolescents (T1D group) and ninety normoglycaemic controls (normoglycaemic group) were included. T1D patients were analysed as a whole and also in subsets of patients with good glycaemic control (glycated hemoglobin concentration ≤7.5%) and with poor glycaemic control (glycated hemoglobin concentration >7.5%). Bone mineral density was assessed by dual energy x-ray absorptiometry. Glycaemic control, renal function and bone markers were also assessed. IGF1, IGF1R and TGFB1 expressions were determined in peripheral blood mononuclear cells by real-time polymerase chain reaction. RESULTS: Patients with T1D showed low bone mineral density and poor glycaemic control. Serum total calcium and urinary albumin-to-creatinine ratio were higher in patients with poor glycaemic control compared to those with good glycemic control (p = 0.003 and p = 0.035, respectively). There was a reduction of IGF1, IGF1R and TGFB1 expressions in the T1D patients and in the subset with poor glycaemic control compared to normoglycaemic controls (p < 0.05). CONCLUSIONS: The decreased IGF1, IGF1R and TGFB1 expressions in the T1D patients, who presented with T1D at an early age, had been diagnosed with T1D for a longer time, had poor glycaemic control and albuminuria may contribute to low bone mineral density. Copyright © 2016 John Wiley & Sons, Ltd.

Association of polymorphisms in IL6 gene promoter region with type 1 diabetes and increased albumin-to-creatinine ratio.

BACKGROUND: Pro-inflammatory cytokines, such as interleukin-6 (IL-6), have been considered as key factors in type 1 diabetes mellitus (T1DM) and diabetic nephropathy, thus, our aim was to investigate the association of IL6-174G>C (rs1800795) and -634C>G (rs1800796) polymorphisms with T1DM susceptibility and diabetic nephropathy. METHODS: These polymorphisms were analyzed in 144 children and adolescents with T1DM and 173 normoglycemic control subjects. Glycemic control, laboratory parameters of kidney function and serum lipids were evaluated. By studying only T1DM patients, we evaluated the polymorphisms associated with relevant biochemical parameters in various genetic models. RESULTS: Type 1 diabetes mellitus patients showed poor glycemic control and albumin-to-creatinine ratio, total cholesterol and LDL-cholesterol levels increased when compared with normoglycemic subjects (p < 0.001, p = 0.004 and p < 0.001, respectively). IL6-174C allele was associated with an increased risk of developing T1DM (OR = 1.53, CI = 1.01-2.31, p = 0.044). In the T1DM group, IL6-174CC carriers showed higher concentrations of glycated hemoglobin (p = 0.029), albumin-to-creatinine ratio (p = 0.021), total cholesterol (p = 0.010), and LDL-cholesterol (p = 0.002), when compared with GG+GC carriers. No association was found for the IL6-634C>G polymorphism. CONCLUSIONS: These results suggest that IL6-174G>C may contribute to T1DM and increased albumin-to-creatinine ratio as well as to poor glycemic control and hyperlipidemia.