Molecular typing of Staphylococcus pseudintermedius canine strains by three commonly used techniques

Staphylococcus pseudintermedius is a newly described species of Staphylococcus regarded as the main causative agent of canine pyoderma [1]. S. pseudintermedius infection was recently described in humans. An important feature of this pathogen is the high genetic identity with two other species of staphylococci, namely S. intermedius and S. delphini, which are included all together in the Staphylococcus Intermedius Group (SIG) [2]. This scenario seriously hampers phenotypic differentiation of these three pathogens. Despite this, only in 2008 was described the first molecular protocol for diagnostic identification of   S. pseudintermedius [3]. The aim of this work was to investigate the presence of different biotypes of S. pseudintermedius obtained from clinically relevant cases of pyoderma in dogs using three molecular methods commonly used to type bacteria: the Ribosomal Spacers Amplification (RSA), the Random Amplification of Polymorphic DNA (RAPD) and the Restriction Fragment Length Polymorphism (RFLP). A total of 46 different strains were included in this work. The application of the RSA technique, which was applied here for the first time, identified the presence of S. pseudintermedius, although it did not allow any differentiation between biotypes. The RAPD assay showed a single cluster that assembles all the interested strains that are grouped in three different sub-clusters (Fig. 1). The RFLP technique showed the most discriminative power, providing the opportunity to clearly identify this bacterium. In conclusion, the use of these three different techniques allows to clearly identify S. pseudintermedius and to observe the presence of different biotypes. In future it could be interesting to couple these results with the determination of the antibiotic resistance in order to verify if certain Multi Drug Resistant strains have particular RSA and RAPD profiles

A Strong Merger Shock in Abell 665

Deep (103 ks) \chandra\ observations of Abell 665 have revealed rich structures in this merging galaxy cluster, including a strong shock and two cold fronts. The newly discovered shock has a Mach number of $M$ = 3.0 $\pm$ 0.6, propagating in front of a cold disrupted cloud. This makes Abell~665 the second cluster where a strong merger shock of $M \approx$ 3 has been detected, after the Bullet cluster. The shock velocity from jump conditions is consistent with (2.7 $\pm$ 0.7) $\times$ 10$^3$ km sec$^{-1}$. The new data also reveal a prominent southern cold front, with potentially heated gas ahead of it. Abell 665 also hosts a giant radio halo. There is a hint of diffuse radio emission extending to the shock at the north, which needs to be examined with better radio data. This new strong shock provides a great opportunity to study the re-acceleration model with the X-ray and radio data combined.Comment: Accepted for publication in ApJ

Characterization of the evolution of noble metal particles in a commercial three‐way catalyst: Correlation between real and simulated ageing

Ageing of automotive catalysts is associated to a loss of their functionality and ultimately to a waste of precious resources. For this reason, understanding catalyst ageing phenomena is necessary for the design of long lasting efficient catalysts. The present work has the purpose of studying in depth all the phenomena that occur during ageing, in terms of morphological modification and deactivation of the active materials: precious metal particles and oxidic support. The topic was deeply investigated using specific methodologies (FT‐IR, CO chemisorption, FE‐ SEM) in order to understand the behavior of metals and support, in terms of their surface properties, morphology and dispersion in the washcoat material. A series of commercial catalysts, aged in different conditions, have been analyzed, in order to find correlations between real and simulated ageing conditions. The characterization highlights a series of phenomena linked to the deactivation of the catalysts. Pd nanoparticles undergo a rapid agglomeration, exhibiting a quick loss of dispersion and of active sites with an increase of particles size. The evolution of the support allows highlighting also the contribution of chemical ageing effects. These results were also correlated with performance tests executed on synthetic gas bench, underlining a good correspondence between vehicle and laboratory aged samples and the contribution of chemical poisoning to vehicle aged ones. The collected data are crucial for the development of accelerated laboratory ageing protocols, which are instrumental for the development and testing of long lasting abatement systems

Morbidity and Mortality in Patients with Idiopathic Pulmonary Fibrosis Undergoing Diagnostic Surgical Lung Biopsy

Morbidity and Mortality In Patients With Idiopathic Pulmonary Fibrosis Undergoing Diagnostic Surgical Lung Biopsy.Previous studies have shown conflicting results about safety of surgical lung biopsy (SLB) in usual interstitial pneumonia (UIP)/idiopathic pulmonary fibrosis (IPF) patients. Study design: we performed a retrospective analysis of all patients who underwent SLB between 2003 and 2013 at the University Hospital of Modena (Italy), to assess morbidity and mortality among patients with UIP/IPF , as compared to patients with other diffuse parenchymal lung diseases (non UIP/IPF). Results: we analyzed 73 patients with (n=29) and without (n=44) UIP/IPF, in which medical history, histology, and survival status were collected. UIP/IPF was diagnosed according to international guidelines. In 59 patients lung biopsy was performed via video-assisted thoracoscopy; in 14 patients thoracotomy was conducted. No intraoperative complications were observed. Postoperative complications were not significantly different between groups and consisted of fever (n=1 in UIP/IPF group; n=4 in non-UIP/IPF group), pneumothorax (n=1 in non-UIP/IPF), pleural effusion (n=1 in non-UIP/IPF), and acute exacerbation (n=1 in both groups, who died). There was no 30-day, 60-day and 90-day mortality. Conclusions: we conclude that surgical lung biopsy can be safely performed in patients with suspected UIP/IPF

Clinical expression of facioscapulohumeral muscular dystrophy in carriers of 1-3 D4Z4 reduced alleles: Experience of the FSHD Italian National Registry

OBJECTIVES: Facioscapulohumeral muscular dystrophy type 1 (FSHD1) has been genetically linked to reduced numbers ( 64 8) of D4Z4 repeats at 4q35. Particularly severe FSHD cases, characterised by an infantile onset and presence of additional extra-muscular features, have been associated with the shortest D4Z4 reduced alleles with 1-3 repeats (1-3 DRA). We searched for signs of perinatal onset and evaluated disease outcome through the systematic collection of clinical and anamnestic records of de novo and familial index cases and their relatives, carrying 1-3 DRA. SETTING: Italy. PARTICIPANTS: 66 index cases and 33 relatives carrying 1-3 DRA. OUTCOMES: The clinical examination was performed using the standardised FSHD evaluation form with validated inter-rater reliability. To investigate the earliest signs of disease, we designed the Infantile Anamnestic Questionnaire (IAQ). Comparison of age at onset was performed using the non-parametric Wilcoxon rank-sum or Kruskal-Wallis test. Comparison of the FSHD score was performed using a general linear model and Wald test. Kaplan-Meier survival analysis was used to estimate the age-specific cumulative motor impairment risk. RESULTS: No patients had perinatal onset. Among index cases, 36 (54.5%) showed the first signs by 10 years of age. The large majority of patients with early disease onset (26 out of 36, 72.2%) were de novo; whereas the majority of patients with disease onset after 10 years of age were familial (16, 53.3%). Comparison of the disease severity outcome between index cases with age at onset before and over 10 years of age, failed to detect statistical significance (Wald test p value=0.064). Of 61 index cases, only 17 (27.9%) presented extra-muscular conditions. Relatives carrying 1-3 DRA showed a large clinical variability ranging from healthy subjects, to patients with severe motor impairment. CONCLUSIONS: The size of the D4Z4 allele is not always predictive of severe clinical outcome. The high degree of clinical variability suggests that additional factors contribute to the phenotype complexity

Inter- and Intraobserver Variation in the Assessment of Preoperative Colostograms in Male Anorectal Malformations: An ARM-Net Consortium Survey

Aim: Male patients with anorectal malformations (ARM) are classified according to presence and level of the recto-urinary fistula. This is traditionally established by a preoperative high-pressure distal colostogram that may be variably interpreted by different surgeons. The aim of this study was to evaluate the inter- and intraobserver variation in the assessment by pediatric surgeons of preoperative colostograms with respect to the level of the recto-urinary fistula. Materials and Methods: Sixteen pediatric surgeons from 14 European centers belonging to the ARM-Net Consortium twice scored 130 images of distal colostograms taken in sagittal projection at a median age of 66 days of life (range: 4–1,106 days). Surgeons were asked to classify the fistula in bulbar, prostatic, bladder-neck, no fistula, and “unclear anatomy” example. Their assessments were compared with the intraoperative findings (kappa) for two scoring rounds with an interval of 6 months (intraobserver variation). Agreement among the surgeons’ scores (interobserver variation) was also calculated using Krippendorff’s alpha. A kappa over 0.75 is considered excellent, between 0.40 and 0.75 fair to good, and below 0.40 poor. Surgeons were asked to score the images in “poor” and “good” quality and to provide their years of experience in ARM treatment. Results: Agreement between the image-based rating of surgeons and the intraoperative findings ranges from 0.06 to 0.45 (mean 0.31). Interobserver variation is higher (Krippendorff’s alpha between 0.40 and 0.45). Years of experience in ARM treatment does not seem to influence the scoring. The mean intraobserver variation between the two rounds is 0.64. Overall, the quality of the images is considered poor. Images categorized as having a good quality result in a statistically significant higher kappa (mean: 0.36 and 0.37 in the first and second round, respectively) than in the group of bad-quality images (mean: 0.25 and 0.23, respectively). Conclusions: There is poor agreement among experienced pediatric colorectal surgeons on preoperative colostograms. Techniques and analyses of images need to be improved in order to generate a homogeneous series of patients and make comparison of outcomes reliable

Elevated TGF \u3b22 serum levels in Emery-Dreifuss Muscular Dystrophy: Implications for myocyte and tenocyte differentiation and fibrogenic processes

Among rare diseases caused by mutations in LMNA gene, Emery-Dreifuss Muscular Dystrophy type 2 and Limb-Girdle muscular Dystrophy 1B are characterized by muscle weakness and wasting, joint contractures, cardiomyopathy with conduction system disorders. Circulating biomarkers for these pathologies have not been identified. Here, we analyzed the secretome of a cohort of patients affected by these muscular laminopathies in the attempt to identify a common signature. Multiplex cytokine assay showed that transforming growth factor beta 2 (TGF \u3b22) and interleukin 17 serum levels are consistently elevated in the vast majority of examined patients, while interleukin 6 and basic fibroblast growth factor are altered in subgroups of patients. Levels of TGF \u3b22 are also increased in fibroblast and myoblast cultures established from patient biopsies as well as in serum from mice bearing the H222P Lmna mutation causing Emery-Dreifuss Muscular Dystrophy in humans. Both patient serum and fibroblast conditioned media activated a TGF \u3b22-dependent fibrogenic program in normal human myoblasts and tenocytes and inhibited myoblast differentiation. Consistent with these results, a TGF \u3b22 neutralizing antibody avoided fibrogenic marker activation and myogenesis impairment. Cell intrinsic TGF \u3b22-dependent mechanisms were also determined in laminopathic cells, where TGF \u3b22 activated AKT/mTOR phosphorylation. These data show that TGF \u3b22 contributes to the pathogenesis of Emery-Dreifuss Muscular Dystrophy type 2 and Limb-Girdle muscular Dystrophy 1B and can be considered a potential biomarker of those diseases. Further, the evidence of TGF \u3b22 pathogenetic effects in tenocytes provides the first mechanistic insight into occurrence of joint contractures in muscular laminopathies

Bowel function and associated risk factors at preschool and early childhood age in children with anorectal malformation type rectovestibular fistula:An ARM-Net consortium study

Background: Outcome of patients operated for anorectal malformation (ARM) type rectovestibular fistula (RVF) is generally considered to be good. However, large multi-center studies are scarce, mostly describing pooled outcome of different ARM-types, in adult patients. Therefore, counseling parents concerning the bowel function at early age is challenging. Aim of this study was to evaluate bowel function of RVF-patients at preschool/early childhood age and determine risk factors for poor functional outcome. Methods: A multi-center cohort study was performed. Patient characteristics, associated anomalies, sacral ratio, surgical procedures, post-reconstructive complications, one-year constipation, and Bowel Function Score (BFS) at 4–7 years of follow-up were registered. Groups with below normal (BFS < 17; subgroups ‘poor’ ≤ 11, and ‘fair’ 11 < BFS < 17) and good outcome (BFS ≥ 17) were formed. Univariable analyses were performed to detect risk factors for outcome. Results: The study included 111 RVF-patients. Median BFS was 16 (range 6–20). The ‘below normal’ group consisted of 61 patients (55.0%). Overall, we reported soiling, fecal accidents, and constipation in 64.9%, 35.1% and 70.3%, respectively. Bowel management was performed in 23.4% of patients. Risk factors for poor outcome were tethered cord and low sacral ratio, while sacral anomalies, low sacral ratio, prior enterostomy, post-reconstructive complications, and one-year constipation were for being on bowel management. Conclusions: Although median BFS at 4–7 year follow-up is nearly normal, the majority of patients suffers from some degree of soiling and constipation, and almost 25% needs bowel management. Several factors were associated with poor bowel function outcome and bowel management. Level of Evidence: Level III