Towards a Long-Read Sequencing Approach for the Molecular Diagnosis of RPGRORF15 Genetic Variants

Sequencing of the low-complexity ORF15 exon of RPGR, a gene correlated with retinitis pigmentosa and cone dystrophy, is difficult to achieve with NGS and Sanger sequencing. False results could lead to the inaccurate annotation of genetic variants in dbSNP and ClinVar databases, tools on which HGMD and Ensembl rely, finally resulting in incorrect genetic variants interpretation. This paper aims to propose PacBio sequencing as a feasible method to correctly detect genetic variants in low-complexity regions, such as the ORF15 exon of RPGR, and interpret their pathogenicity by structural studies. Biological samples from 75 patients affected by retinitis pigmentosa or cone dystrophy were analyzed with NGS and repeated with PacBio. The results showed that NGS has a low coverage of the ORF15 region, while PacBio was able to sequence the region of interest and detect eight genetic variants, of which four are likely pathogenic. Furthermore, molecular modeling and dynamics of the RPGR Glu-Gly repeats binding to TTLL5 allowed for the structural evaluation of the variants, providing a way to predict their pathogenicity. Therefore, we propose PacBio sequencing as a standard procedure in diagnostic research for sequencing low-complexity regions such as RPGRORF15, aiding in the correct annotation of genetic variants in online databases

Is there a correlation between nasal septum deviation and maxillary transversal deficiency? A retrospective study on prepubertal subjects

Introduction: Deviated nasal septum may cause a reduction of the nasal airflow, thus, during the craniofacial development, a reduced nasal airflow could originate a chronic mouth-breathing pattern, related with moderate to severe maxillary constriction. The aim of this retrospective study is to analyze the correlation between maxillary transverse deficiency and nasal septum deviation.Methods: Frontal cephalograms were performed on 66 posterior-anterior radiographs of subjects (34M, 32F; mean age 9.95 +/- 2.50 years) with maxillary transverse deficiency and on a control group of 31 posterior-anterior radiographs of subjects (13M, 18F; 9.29 +/- 2.08 years). Angular parameters of the nasal cavities were recorded and compared between the two groups using a Student's t-test.Results: Generally all the parameters are very similar between the two groups except for the ASY angle that differs for about the 27%; anyway the Student's t-test showed no statistically significant differences between the two groups (mostly p > 0.20).Conclusions: This study failed to show an association between transverse maxillary deficiencies and nasal septum deviations. Moreover, no significant differences were found between the mean nasal cavities dimensions in subjects with transverse maxillary deficiency and the control group. (C) 2016 Elsevier Ireland Ltd. All rights reserved

Influence of the mandibular position on the active cervical range of motion of healthy subjects analyzed using an accelerometer

Objective: The aim of this study is to analyze the influence of the mandibular positions (habitual rest position, habitual maximum intercuspation, habitual maximum intercuspation with clenching, and mandibular position with cotton rolls) on the active cervical range of motion (ROM) (flexion-extension, lateroflexions, rotations) using an accelerometer in a sample of healthy subjects. Methods: A total of 21 (14 M, 7 F) healthy volunteers aged from 18 to 27 years (mean age 23.88 ± 2.34 years; mean weight 67.86 ± 11.38 kg; mean height 172.52 ± 9.00 cm) underwent a cervical range of movement examination using a 9-axis accelerometer. A one-way ANOVA analysis was performed in order to statistically evaluate the effective influence of the mandibular position on the recorded parameters. Results: The analysis showed no statistically significant differences (all p-values > 0.1) with variations smaller than three degrees among the different mandibular positions. Discussion: The mandibular position seems to have no influence on the active cervical ROM in healthy subjects. Further studies are needed to assess the usefulness of the accelerometer in the cervical analysis of temporomandibular disorder subjects

Predizione precoce di malattie cardiovascolari in pazienti con trapianto di rene

Razionale : Il rischio di malattie cardiovascolari (CVD) è elevato nel trapianto di rene (Tx-rene). Questo studio ha analizzato in pazienti con Tx-rene il potere predittivo di CVD fornito da informazioni disponibili pre-trapianto o nei primi sei mesi post-trapianto. Casistica e Metodi: Coorte target erano pazienti con Tx-rene in età adulta (≥ 18 anni) effettuato nel periodo 2005-10. Obiettivo dell’analisi era l’incidenza post-trapianto fino al 2015 di CVD (malattie coronariche, malattie cerebrovascolari, arteriopatie periferiche) oppure di evento competitivo composito (follow-up interrotto, insufficienza del trapianto, morte per malattia non-cardiovascolare con trapianto funzionante). Il potere predittivo di CVD è stato indagato per 34 variabili usando modelli multi-variati di regressione di Cox con inclusione di rischio competitivo. Risultati: La coorte dello studio consisteva di 629 pazienti. Il follow-up post-trapianto variava da 0.28 a 10.00 anni (mean±SD = 7.30±3.10). Primo evento incidente era una CVD in 103 pazienti ed un evento competitivo in 146 pazienti. In modelli multi-variati limitati alle sole variabili pre-trapianto, predittori indipendenti di CVD erano nefropatia diabetica (hazard ratio, HR = 6.63, 95%CI = 1.81/24.35), CVD pre-trapianto (4.87, 2.84/8.35), età al trapianto ≥ 45 anni (2.98, 1.83/4.87), sesso maschile (1.68, 1.06/2.66) e durata dialisi pre-trapianto ≥ 5 anni (1.52, 1.02/2.27). In modelli multi-variati per variabili pre-trapianto e post-trapianto insieme, la sola variabile post-trapianto con potere predittivo indipendente era evidenza a sei mesi dal traapianto di eGFR <60 mL/min x 1.73 m2 (1.75, 1.11/2.77). Da sole, nefropatia diabetica, dialisi pre-trapianto ≥ 5 anni, e CVD pre-trapianto predicevano il 91% delle CVD incidenti. L’aggiunta di altre variabili non aumentava significativamente questa percentuale. Conclusioni: I dati indicano che semplici informazioni disponibili al trapianto hanno un elevato potere predittivo per CVD post-trapianto. I dati non supportano un ruolo preponderante di fattori post-trapianto nel predire o determinare l’incidenza di malattie CVD post-trapianto

Gnathological features in growing subjects

Aim of this study was to evaluate the prevalence of temporomandibular disorders (TMD) in a sample of consecutive subjects. Materials and methods: TMDs were recorded in a sample of 580 subjects (279 M, 301 F; mean age: 13.4y). For each subject a case history was compiled to evaluate the social and demographic parameters. An extraoral exam was effected to point out the face proportions, and an intraoral exam was performed to analyze dental occlusion, mandibular deviation during opening, presence of cross-bites, overjet and overbite. A functional exam was carried out to evaluate mandibular movements and to find joint sounds and myofascial pain. The sample was divided into 6 groups according to the: gender, age (ages 6y- 11y and 12y-16y), Angle Dental Class, cross-bite, midline deviation and chewing side. For this investigation latex gloves, a millimeter calipers (precision 0,01 mm) and a phonendoscope were used. The percentages of signs and symptoms were compared using the ?2-test with Yates correction to determine the differences among the groups for the rates of TMDs, reduced opening/lateral/protrusive movements, and myofascial pain. Results: The prevalence of TMDs in the total sample was 13,9%. Among 6y-11y subjects the percentage of TMD was 7,3% while it was 16,1% among 12y-16y subjects (?2=1.634;; p=0.201). Females showed a percentage of 16,6% of TMDs while males one of 10,8% (?2=0.556;; p=0.456). According to angle malocclusion, the prevalence was 14% in subjects with Class I malocclusion, 15% in sample with Class II and 9% in patients with Class III (?2=0.540;; p=0.763). According to presence or absence of crossbite, prevalence of TMD signs and symptoms was 13,8% among subjects without crossbite and 14,3% among subjects with crossbite, with no significant difference between the two subgroups (?2= 0,047619; p=0.050). In relation of midline deviation, prevalence of TMDs was 15% in subjects without deviation, 15,8% in functional deviation subjects and 4,7% in anatomic deviation ones (?2=1.555;; p=0.05). Prevalence of TMDs was 12,6% in subjects with bilateral chewing and 28% in unilateral chewing. Conclusions: TMDs seem to be not associated to age, to gender, Angle Class, cross-bite and chewing side

Predizione precoce di malattie cardiovascolari in pazienti con trapianto di rene

Razionale : Il rischio di malattie cardiovascolari (CVD) è elevato nel trapianto di rene (Tx-rene). Questo studio ha analizzato in pazienti con Tx-rene il potere predittivo di CVD fornito da informazioni disponibili pre-trapianto o nei primi sei mesi post-trapianto. Casistica e Metodi: Coorte target erano pazienti con Tx-rene in età adulta (≥ 18 anni) effettuato nel periodo 2005-10. Obiettivo dell’analisi era l’incidenza post-trapianto fino al 2015 di CVD (malattie coronariche, malattie cerebrovascolari, arteriopatie periferiche) oppure di evento competitivo composito (follow-up interrotto, insufficienza del trapianto, morte per malattia non-cardiovascolare con trapianto funzionante). Il potere predittivo di CVD è stato indagato per 34 variabili usando modelli multi-variati di regressione di Cox con inclusione di rischio competitivo. Risultati: La coorte dello studio consisteva di 629 pazienti. Il follow-up post-trapianto variava da 0.28 a 10.00 anni (mean±SD = 7.30±3.10). Primo evento incidente era una CVD in 103 pazienti ed un evento competitivo in 146 pazienti. In modelli multi-variati limitati alle sole variabili pre-trapianto, predittori indipendenti di CVD erano nefropatia diabetica (hazard ratio, HR = 6.63, 95%CI = 1.81/24.35), CVD pre-trapianto (4.87, 2.84/8.35), età al trapianto ≥ 45 anni (2.98, 1.83/4.87), sesso maschile (1.68, 1.06/2.66) e durata dialisi pre-trapianto ≥ 5 anni (1.52, 1.02/2.27). In modelli multi-variati per variabili pre-trapianto e post-trapianto insieme, la sola variabile post-trapianto con potere predittivo indipendente era evidenza a sei mesi dal traapianto di eGFR <60 mL/min x 1.73 m2 (1.75, 1.11/2.77). Da sole, nefropatia diabetica, dialisi pre-trapianto ≥ 5 anni, e CVD pre-trapianto predicevano il 91% delle CVD incidenti. L’aggiunta di altre variabili non aumentava significativamente questa percentuale. Conclusioni: I dati indicano che semplici informazioni disponibili al trapianto hanno un elevato potere predittivo per CVD post-trapianto. I dati non supportano un ruolo preponderante di fattori post-trapianto nel predire o determinare l’incidenza di malattie CVD post-trapianto

EARLY PREDICTION OF CARDIOVASCULAR DISEASE IN KIDNEY TRANSPLANT RECIPIENTS

Cardiovascular disease (CVD) is frequent in kidney transplant (Tx). This study investigated CVD prediction in kidney Tx by information available pre-Tx or within six months post-Tx. Study cohort consisted of 629 patients with Tx in 2005-10 and with adult age at Tx. Endpoint was the incidence up to 2015 of CVD (coronary heart disease, cerebrovascular disease, peripheral artery disease). Graft failure, non-CVD death with functioning graft, and loss to follow-up were considered competing events. CVD prediction was investigated for thirty-four variables using competing-risks regression. Follow-up range was 0.28-10.00 years (mean±SD= 7.30±3.10). First incident event was CVD in 103 patients and competing events in 146 patients. In multi-variable model for pre-Tx variables only, CVD predictors were male sex (hazard ratio= 1.68, 95%CI= 1.06/2.66), diabetic nephropathy (6.63, 1.81/24.35), pre-Tx dialysis for ≥5 years (1.52, 1.02/2.27), pre-Tx CVD (4.87, 2.84/8.35), age at Tx ≥45 years (2.98, 1.83/4.87). In model for pre-Tx and post-Tx variables together, the sole post-Tx CVD predictor was estimated glomerular filtration rate <60 mL/min at the 6-month visit (1.75, 1.11/2.77). Diabetic nephropathy, pre-Tx dialysis, pre-Tx CVD, and age at Tx predicted 91.2% of incident CVD. Early available information effectively predicted CVD in kidney Tx independent of competing events

Predizione precoce di malattie cardiovascolari in pazienti con trapianto di rene

Razionale : Il rischio di malattie cardiovascolari (CVD) è elevato nel trapianto di rene (Tx-rene). Questo studio ha analizzato in pazienti con Tx-rene il potere predittivo di CVD fornito da informazioni disponibili pre-trapianto o nei primi sei mesi post-trapianto. Casistica e Metodi: Coorte target erano pazienti con Tx-rene in età adulta (≥ 18 anni) effettuato nel periodo 2005-10. Obiettivo dell’analisi era l’incidenza post-trapianto fino al 2015 di CVD (malattie coronariche, malattie cerebrovascolari, arteriopatie periferiche) oppure di evento competitivo composito (follow-up interrotto, insufficienza del trapianto, morte per malattia non-cardiovascolare con trapianto funzionante). Il potere predittivo di CVD è stato indagato per 34 variabili usando modelli multi-variati di regressione di Cox con inclusione di rischio competitivo. Risultati: La coorte dello studio consisteva di 629 pazienti. Il follow-up post-trapianto variava da 0.28 a 10.00 anni (mean±SD = 7.30±3.10). Primo evento incidente era una CVD in 103 pazienti ed un evento competitivo in 146 pazienti. In modelli multi-variati limitati alle sole variabili pre-trapianto, predittori indipendenti di CVD erano nefropatia diabetica (hazard ratio, HR = 6.63, 95%CI = 1.81/24.35), CVD pre-trapianto (4.87, 2.84/8.35), età al trapianto ≥ 45 anni (2.98, 1.83/4.87), sesso maschile (1.68, 1.06/2.66) e durata dialisi pre-trapianto ≥ 5 anni (1.52, 1.02/2.27). In modelli multi-variati per variabili pre-trapianto e post-trapianto insieme, la sola variabile post-trapianto con potere predittivo indipendente era evidenza a sei mesi dal traapianto di eGFR <60 mL/min x 1.73 m2 (1.75, 1.11/2.77). Da sole, nefropatia diabetica, dialisi pre-trapianto ≥ 5 anni, e CVD pre-trapianto predicevano il 91% delle CVD incidenti. L’aggiunta di altre variabili non aumentava significativamente questa percentuale. Conclusioni: I dati indicano che semplici informazioni disponibili al trapianto hanno un elevato potere predittivo per CVD post-trapianto. I dati non supportano un ruolo preponderante di fattori post-trapianto nel predire o determinare l’incidenza di malattie CVD post-trapianto