Analysis of LMNB1 Duplications in Autosomal Dominant Leukodystrophy Provides Insights into Duplication Mechanisms and Allele‐Specific Expression

Autosomal dominant leukodystrophy (ADLD) is an adult onset demyelinating disorder that is caused by duplications of the lamin B1 (LMNB1) gene. However, as only a few cases have been analyzed in detail, the mechanisms underlying LMNB1 duplications are unclear. We report the detailed molecular analysis of the largest collection of ADLD families studied, to date. We have identified the minimal duplicated region necessary for the disease, defined all the duplication junctions at the nucleotide level and identified the first inverted LMNB1 duplication. We have demonstrated that the duplications are not recurrent; patients with identical duplications share the same haplotype, likely inherited from a common founder and that the duplications originated from intrachromosomal events. The duplication junction sequences indicated that nonhomologous end joining or replication‐based mechanisms such fork stalling and template switching or microhomology‐mediated break induced repair are likely to be involved. LMNB1 expression was increased in patients’ fibroblasts both at mRNA and protein levels and the three LMNB1 alleles in ADLD patients show equal expression, suggesting that regulatory regions are maintained within the rearranged segment. These results have allowed us to elucidate duplication mechanisms and provide insights into allele‐specific LMNB1 expression levels.     

EGFR-Targeted Therapy for Non-Small Cell Lung Cancer: Focus on EGFR Oncogenic Mutation

The two essential requirements for pathologic specimens in the era of personalized therapies for non-small cell lung carcinoma (NSCLC) are accurate subtyping as adenocarcinoma (ADC) versus squamous cell carcinoma (SqCC) and suitability for EGFR molecular testing, as well as for testing of other oncogenes such as EML4-ALK and KRAS. Actually, the value of EGFR expressed in patients with NSCLC in predicting a benefit in terms of survival from treatment with an epidermal growth factor receptor targeted therapy is still in debate, while there is a convincing evidence on the predictive role of the EGFR mutational status with regard to the response to tyrosine kinase inhibitors (TKIs).This is a literature overview on the state-of-the-art of EGFR oncogenic mutation in NSCLC. It is designed to highlight the preclinical rationale driving the molecular footprint assessment, the progressive development of a specific pharmacological treatment and the best method to identify those NSCLC who would most likely benefit from treatment with EGFR-targeted therapy. This is supported by the belief that a rationale for the prioritization of specific regimens based on patient-tailored therapy could be closer than commonly expected.     

Dentinal tubule penetration and root canal cleanliness following ultrasonic activation of intracanal‐heated sodium hypochlorite

This study investigated the effect of ultrasonic activation of intracanal‐heated sodium hypochlorite (NaOCl) on its dentinal tubular penetration and root canal cleanliness in vitro. In experiment 1, mandibular premolars were randomly allocated to three groups (n = 8): group A, ultrasonic activation; group B, ultrasonic activation of intracanal‐heated NaOCl and group C, syringe‐and‐needle irrigation. Penetration of the fluorescent‐labelled NaOCl was investigated using light microscopy. In experiment 2, mandibular premolars were randomly allocated to group B or C (n = 10), for histological analysis of the remaining pulp tissue and debris. Data were statistically analysed using Kruskal–Wallis and Mann–Whitney tests (P = 0.05). The highest penetration of NaOCl was observed in group B, followed by group A (P < 0.05). Group B showed significantly less amount of debris than group C (P < 0.05). Dentinal tubule penetration of NaOCl and root canal cleanliness were significantly improved by ultrasonic activation of intracanal‐heated NaOCl.     

Effects of Dry Needling on Muscle Stiffness in Latent Myofascial Trigger Points: A Randomized Controlled Trial

The aim of this study was to analyze the effects of dry needling (DN) in upper trapezius latent trigger points (LTrPs) on muscle stiffness. A total of 51 recreational physically active subjects with LTrPs in the upper trapezius volunteered to participate and were randomly divided into a DN-group (n = 27) and a sham-DN group (n = 24). Volunteers received 1-session of DN or placebo treatment. Muscle stiffness, measured with strain and shear-wave elastography, pressure pain threshold (PPT), post-needling soreness, and muscle thickness were evaluated before treatment, and at 30-min, 24-hours, and 72-hours follow-up after treatment. The DN-group showed lower values from baseline for muscle stiffness measured with shear-wave elastrography at 24-hours (from 44.44 ± 15.97 to 35.78 ± 11.65 kpa; P < .01) and at 72-hours (35.04 ± 12.61 kpa; P < .01) and with strain elastography at 72-hours (from 1.75 ± 0.50 to 1.36 ± 0.40 AU; P < .01). The DN-group showed higher values of PPT than the sham-DN group at 72-hours (4.23 ± 0.75 vs. 5.19 ± 1.16 kg/cm²; P < .05). There was a progressive decrease in post-needling soreness compared to pain during needling of 33.13 ± 21.31% at 30-min, 80.92 ± 10.06% at 24-hours, and a total decrease in post-needling soreness in all participants at 72-hours. DN therapy is effective in reducing short-term muscle stiffness and increasing the PPT in volunteers with LTrPs in the upper trapezius after a treatment session.PerspectiveThis study found that one session of DN intervention in latent trigger points of the upper trapezius muscle reduced muscle stiffness and the pressure pain threshold for the dry needling group compared to the sham dry needling group.     

Three-Dimensional Laparoscopic Sleeve Gastrectomy: Improved Patient Safety and Surgeon Convenience

One of the aims of laparoscopic surgery is to improve upon the results obtained by open surgery. This clearly appears to have been achieved in bariatric surgery. Two-dimensional (2-D) systems have been used to date, though new 3-dimensional (3-D) technologies have been introduced in an attempt to improve surgeon vision and thus increase the safety of the surgical techniques. Sixty obese patients underwent sleeve gastrectomy using a device equipped with 3-D optics allowing surgery to be viewed by the surgeon in 3 dimensions by using a specific monitor and wearing appropriate glasses. The mean patient age was 48.1 years. The mean weight was 114 kg (range, 92–172), with a mean body mass index (BMI) of 44 ± 5.21 kg/m². All surgeries were performed using the 3-D system, with a mean surgical time of 71 ± 49.6 minutes and a mean hospital stay of 3.0 ± 1.2 days. Only 1 intraoperative complication was recorded: retroperitoneal bleeding on insertion of the optical trocar. Over a mean follow-up period of 12 months, the mean body weight of the patients was 88 kg (range, 71–121), with a BMI of 30.56 ± 3.98 kg/m² and a percentage excess weight loss of 68.14% ± 7.89%. There was clear improvement of both the blood pressure and glucose levels. Three-dimensional sleeve gastrectomy is safe, viable, and fully reproducible compared with 2-D surgery, improving visualization of the surgical field, safety, and surgeon convenience. Randomized studies involving larger patient samples are needed for the comparison of results.Key words: 3-D laparoscopy, Bariatric surgery, Metabolic surgeryIn recent years, laparoscopic surgery has become clearly indicated in bariatric and metabolic surgery. This surgical technique has made it possible to reduce morbidity and mortality in comparison with conventional open surgery, and to shorten hospital stay. In addition to the usual 5-trocar technique, laparoscopic surgery has also produced satisfactory results with the single-port approach, which is regarded as less invasive and affords equivalent outcomes and improved aesthetic results.More recently, the possibility of using three-dimensional (3-D) surgery has been considered. The film Avatar (James Cameron, 2006) represented a starting point for a number of multinational medical engineering companies who launched projects that now allow us to use 3-D laparoscopic systems. This new technology offers the perception of depth, which was not previously available.Sleeve gastrectomy is one of the surgical techniques presently used to treat obesity. It was initially regarded as first-step surgery in patients with a high body mass index (BMI), before performing biliopancreatic diversion. Sleeve gastrectomy is currently considered to be a restrictive technique that can be used as a sole option in morbid obesity, even when accompanied by diabetes mellitus, as contemplated by the International Sleeve Gastrectomy Expert Panel Consensus of 2011.Three-dimensional laparoscopic sleeve gastrectomy requires 5 ports, like the two-dimensional (2-D) laparoscopic technique, and one of them must be enlarged to extract the sectioned stomach. However, 3-D visualization is very useful in all the surgical steps, as it improves spatial orientation and can increase the safety of the surgical technique.The present study analyzes the results of 3-D laparoscopic sleeve gastrectomy after a short 1-year follow-up period, with a view to evaluating its safety and feasibility in patients with severe or morbid obesity and type 2 diabetes.     

Evaluación multimodal de la degeneración estructural de válvulas percutáneas en el seguimiento a largo plazo

Introduction and objectivesWe assessed the long-term hemodynamic performance of transcatheter heart valve (THV) by paired transthoracic echocardiography (TTE), and the incidence, characteristics and factors associated with THV structural valve degeneration (SVD).MethodsA total of 212 patients who underwent transcatheter aortic valve replacement and had a potential follow-up > 5 years with at least 1 TTE ≥ 1-year postprocedure were included. All patients had a TTE at 1 to 5 years and 36 had another one at 6 to 10 years. SVD was defined as subclinical (increase > 10 mmHg in mean transvalvular gradient +  decrease > 0.3 cm² in valve area and/or new-onset mild or moderate aortic regurgitation) and clinically relevant (increase > 20 mmHg in mean transvalvular gradient + decrease > 0.6 cm² in valve area and/or new-onset moderate-to-severe aortic regurgitation). Fifteen patients had a transesophageal echocardiography at the time of SVD diagnosis, and 85 an opportunistic computed tomography examination at 1 (0.5-2) years.ResultsTransvalvular mean gradient increased and valve area decreased over time (P < .01). At 8 years of follow-up, SVD occurred in 30.2% of patients (clinically relevant: 9.3%). Transesophageal echocardiography revealed thickened and reduced-mobility leaflets in 80% and 73% of SVD cases, respectively. No baseline or procedural factors were associated with SVD. THV underexpansion (3.5%) or eccentricity (8.2%) had no impact on valve hemodynamics/SVD at follow-up.ConclusionsA gradual THV hemodynamic deterioration occurred throughout a 10-year period, leading to SVD in ～30% of patients (clinically relevant in < 10%). Leaflet morphology/mobility were frequently impaired in SVD cases, but THV geometry did not influence valve hemodynamics or SVD.     

Comparison of an interferon-gamma assay with tuberculin skin testing for detection of tuberculosis (TB) infection in patients with rheumatoid arthritis in a TB-endemic population

OBJECTIVE: Tuberculosis (TB) in patients with rheumatoid arthritis (RA) undergoing treatment with anti-tumor necrosis factor (TNF) agents is commonly the result of reactivation of latent TB infection (LTBI); detection and treatment of LTBI is essential before treatment with anti-TNF agents. We reported previously that the tuberculin skin test (TST) is inaccurate for diagnosis of LTBI in patients with RA. Here, we compare the prevalence of LTBI in RA patients and matched controls according to positive TST and QuantiFeron-TB Gold In-Tube version (QFT) results and determine their agreement. METHODS: A cross-sectional study of 101 RA patients and 93 controls was conducted in Lima, Perú, where the prevalence of LTBI in the general population has been estimated to be 68%. Blood was drawn for QFT assay followed by TST using 2-TU of RT 23 purified protein derivative. TST was deemed positive at >or= 5 mm for RA patients and >or= 10 mm for controls. RESULTS: There were no significant differences between RA patients and controls for age, sex, bacillus Calmette-Guérin vaccination, or history of or contact with TB. 88% of patients had active RA disease and 2 (1.9%) patients had indeterminate QFT results. The number of subjects testing positive with the QuantiFeron assay was comparable between patients and controls (44.6% vs 59.1%, respectively), whereas the TST detected significantly less LTBI among RA patients (26.7%) than controls (65.6%). Thus, the rate of LTBI in RA patients represented 75% and 41% of the rate in their controls using QFT or TST, respectively (p = 0.008). Poor agreement between TST and QFT was seen in RA patients, but in controls, good agreement was observed between these tests. CONCLUSION: In a TB-endemic population, the QuantiFeron-TB Gold In-Tube assay seemed to be a more accurate test for detection of LTBI in RA patients compared with the TST, and may potentially improve the targeting of prophylactic therapy before treatment with anti-TNF agents.     

Intravenous immunoglobulin treatment for acute fulminant inflammatory cardiomyopathy: series of six patients and review of literature

BACKGROUND:

Although an autoimmune mechanism has been postulated for myocarditis and acute-onset inflammatory dilated cardiomyopathy (DCM), immunomodulatory treatment strategies are still under investigation.

METHODS AND RESULTS:

The clinical data of six patients with acute inflammatory DCM referred for evaluation for possible heart transplantation were reviewed. All patients were admitted with acute congestive heart failure and severely impaired left ventricular (LV) function and were treated with high-dose (2 g/kg) intravenous immunoglobulin (IVIG). The diagnosis of acute inflammatory DCM was based on recent onset of congestive heart failure (New York Heart Association functional class III or IV) with severely depressed LV ejection fraction ([LVEF] 30% or lower) occurring shortly after viral-like illness. All patients had inflammation on endomyocardial biopsy or elevated cardiac enzymes, as well as a normal coronary angiogram. All patients were in New York Heart Association class I or II at the time of hospital discharge. The mean LVEF improved from 21.7±7.5% at baseline to 50.3±8.6% at discharge (P=0.005). Four patients had complete recovery (LVEF 50% or higher) and two patients had partial LV recovery. Patients were followed for a median 13.2 months (range two to 24 months) and had a mean LVEF of 53±6% (P not significant versus LVEF at discharge).

CONCLUSIONS:

Therapy with intravenous high-dose IVIG may be a potentially useful treatment in selected patients if given early in the course of acute fulminant inflammatory DCM. A randomized, prospective trial is warranted to prove the real benefit of IVIG in this patient population.