High-Resolution Identification of Multiple Salmonella Serovars in a Single Sample by Using CRISPR-SeroSeq

Salmonella enterica is represented by \u3e2,600 serovars that can differ in routes of transmission, host colonization, and in resistance to antimicrobials. S. enterica is the leading bacterial cause of foodborne illness in the United States, with well-established detection methodology. Current surveillance protocols rely on the characterization of a few colonies to represent an entire sample; thus, minority serovars remain undetected. Salmonella contains two CRISPR loci, CRISPR1 and CRISPR2, and the spacer contents of these can be considered serovar specific. We exploited this property to develop an amplicon-based and multiplexed sequencing approach, CRISPR-SeroSeq (serotyping by sequencing of the CRISPR loci), to identify multiple serovars present in a single sample. Using mixed genomic DNA from two Salmonella serovars, we were able to confidently detect a serovar that constituted 0.01% of the sample. Poultry is a major reservoir of Salmonella spp., including serovars that are frequently associated with human illness, as well as those that are not. Numerous studies have examined the prevalence and diversity of Salmonella spp. in poultry, though these studies were limited to culture-based approaches and therefore only identified abundant serovars. CRISPR-SeroSeq was used to investigate samples from broiler houses and a processing facility. Ninety-one percent of samples harbored multiple serovars, and there was one sample in which four different serovars were detected. In another sample, reads for the minority serovar comprised 0.003% of the total number of Salmonella spacer reads. The most abundant serovars identified were Salmonella enterica serovars Montevideo, Kentucky, Enteritidis, and Typhimurium. CRISPR-SeroSeq also differentiated between multiple strains of some serovars. This high resolution of serovar populations has the potential to be utilized as a powerful tool in the surveillance of Salmonella species

Regional Dominant Frequency: A New Tool for Wave Break Identification During Atrial Fibrillation

Cardiac mapping systems are based on the time/frequency feature analyses of intracardiac electrograms recorded from individual bipolar/unipolar electrodes. Signals from each electrode are processed independently. Such approaches fail to investigate the interrelationship between simultaneously recorded channels of any given mapping catheter during atrial fibrillation (AF). We introduce a novel signal processing technique that reflects regional dominant frequency (RDF) components. We show that RDF can be used to identify and characterize variation and disorganization in wavefront propagation- wave breaks. The intracardiac electrograms from the left atrium of 15 patients were exported to MATLAB and custom software employed to estimate RDF and wave break rate (WBR). We observed a heterogeneous distribution of both RDF and WBR; the two measures were weakly correlated (0.3; p < 0.001). We identified locations of AF or atrial tachycardia (ATach) termination and later compared offline with RDF and WBR maps. We inspected our novel metrics for associations with AF termination sites. Areas associated with AF termination demonstrated high RDF and low WBR (↑RDF,↓WBR). These sites were present in 14 of 15 patients (mean 2.6 ± 1.2 sites per patient; range, 1–4 sites), 43% situated within the pulmonary veins. In nine patients where AF terminated to sinus rhythm (6) or ATach (3), post-hoc analysis demonstrated all ↑RDF,↓WBR sites were ablated and correlated with AF termination sites. The proposed RDF signal processing tools can be used to identify and quantify wave break, and the combined use of these two novel metrics can aid characterization of AF. Further prospective studies are warranted

How to treat a patient with T1 high-grade disease and no tumour on repeat transurethral resection of the bladder?

A relatively young (64-yr old) long-term heavy smoker but otherwise very healthy man is diagnosed with a primary unifocal left-side tumour (urothelial, T1 high grade), but no lymphovascular invasion and no variant histology. We discuss whether treatment with intravesical bacillus Calmette-Guérin vaccine will be sufficient or early radical cystectomy is at least equally preferred regarding patient benefit, safety, and quality of life. Patient summary A patient with a single high-grade T1 bladder tumour without aggressive features (eg, lymphovascular invasion or variant tumour aspects) will be adequately treated with bacillus Calmette-Guérin intravesical therapy delivered into the bladder, followed by 3 yr of maintenance. However, all decisions should be taken with the patient in a shared decision-making process, including a discussion regarding removal of the bladder

The effects of suppressing inflammation by tofacitinib may simultaneously improve glycaemic parameters and inflammatory markers in rheumatoid arthritis patients with comorbid type 2 diabetes: a proof-of-concept, open, prospective, clinical study

Background: A consistent connection has been increasingly reported between rheumatoid arthritis (RA), insulin resistance (IR), and type 2 diabetes (T2D). The β-cell apoptosis induced by pro-inflammatory cytokines, which could be exaggerated in the context of RA, is associated with increased expression pro-apoptotic proteins, which is dependent on JAnus Kinase/Signal Transducer and Activator of Transcription (JAK/STAT) activation. On these bases, we aimed to evaluate if the administration of tofacitinib, a potent and selective JAK inhibitor, could simultaneously improve glycaemic parameters and inflammatory markers in patients with RA and comorbid T2D. Methods: The primary endpoint was the change in the 1998-updated homeostatic model assessment of IR (HOMA2-IR) after 6 months of treatment with tofacitinib in RA patients with T2D. Consecutive RA patients with T2D diagnosis were included in this proof-of-concept, open, prospective, clinical study, which was planned before the recent emergence of safety signals about tofacitinib. Additional endpoints were also assessed regarding RA disease activity and metabolic parameters. Results: Forty consecutive RA patients with T2D were included (female sex 68.9%, mean age of 63.4 ± 9.9 years). During 6-month follow-up, a progressive reduction of HOMA2-IR was observed in RA patients with T2D treated with tofacitinib. Specifically, a significant effect of tofacitinib was shown on the overall reduction of HOMA2-IR (β = − 1.1, p = 0.019, 95%CI − 1.5 to − 0.76). Also, HOMA2-β enhanced in these patients highlighting an improvement of insulin sensitivity. Furthermore, although a longer follow-up is required, a trend in glycated haemoglobin reduction was also recorded. The administration of tofacitinib induced an improvement in RA disease activity, and a significant reduction of DAS28-CRP and SDAI was observed; 76.8% of patients achieved a good clinical response. In this study, no major adverse events (AEs) were retrieved without the identification of new safety signals. Specifically, no life-threatening AEs and cardiovascular and/or thromboembolic events were recorded. Conclusions: The administration of tofacitinib in RA with T2D led to a simultaneous improvement of IR and inflammatory disease activity, inducing a “bidirectional” benefit in these patients. However, further specific designed and powered studies are warranted to entirely evaluate the metabolic effects of tofacitinib in RA patients with T2D

Celecoxib as an Adjuvant to Fluvoxamine in Moderate to Severe Obsessive-compulsive Disorder: A Double-blind, Placebo-controlled, Randomized Trial

Introduction: A growing body of evidence implicates inflammatory cascades in the pathophysiology of obsessive-compulsive disorder (OCD), making this pathway a target for development of novel treatments. Methods: 50 outpatients with moderate to severe OCD participated in the trial, and underwent 10 weeks of treatment with either celecoxib (200 mg twice daily) or placebo as an adjuvant to fluvoxamine. Participants were investigated using Yale-Brown Obsessive Compulsive Scale (Y-BOCS). The main outcome measure was to assess the efficacy of celecoxib in improving the OCD symptoms. Results: General linear model repeated measures demonstrated significant effect for time � treatment interaction on the Y-BOCS total scores F (1.38, 66.34)=6.91, p=0.005. Kaplan-Meier estimation with log-rank test demonstrated significantly more rapid response in the celecoxib group than the placebo group (p<0.001). There was no significant difference in adverse event frequencies between the groups. Discussion: The results of the current study suggest that celecoxib could be a tolerable and effective adjunctive treatment for more rapid and more satisfying improvements in OCD symptoms. © Georg Thieme Verlag KG Stuttgart, New York

Effect of intensive neuromuscular electrical stimulation on chronic neck pain: A case report

© Nova Science Publishers, Inc. Chronic neck pain is a relatively common problem that can interfere with daily activities, and it is often experienced following musculoskeletal injuries. To identify the impact of intensive neuromuscular electrical stimulation (INES) for reducing chronic neck pain in a 21-year-old female athlete, following a traumatic sports injury, which occurred two years earlier. A treatment package including three separate sessions of intensive neuromuscular electrical stimulation and exercise therapy were prescribed. Outcomes measurements were short form McGill pain questionnaire (SF-MPQ), visual analogue scale (VAS), and the neck disability index (NDI). Measurements were performed at baseline, following the intervention, and three months later. Following our intervention; VAS score decreased from 6/10 to 3/10, and 1/10 after three months; and NDI decreased from 54/100 to 18/100, and 10/100 after three months. A combination of INES and resistance training significantly reduced neck pain after three months in a female gymnast. Further research is required to determine the effectiveness of this combination of treatments in larger cohorts with more diffuse musculoskeletal conditions

Comparison of risk factors related to intraventricular hemorrhage between preterm infants born after normal and in vitro fertilization conceptions

Objectives We aimed to compare the level of significance of risk factors related Intraventricular hemorrhage (IVH) between preterm infants born after IVF and non-IVF conceptions. Materials & Methods This historical cohort study was done in four Iranian Hospitals in 2013-2014. Overall, 155 preterm newborns were divided into case (IVF) and control (normal conception) groups. Both groups� demographic data were extracted and recorded. The incidence of IVH and its grades were compared between case and control groups. Significant related risk factors were also considered. Results No differences were observed between 2 groups except for gestational age and mode of delivery. The incidence of IVH especially grades II and III were significantly higher in the case group (P=0.003). Results showed no correlations between Gestational age (GA), birth weight and number of gestations with the incidence of IVH in the case group (0.059, 0.85 and 0.49, respectively). On the other hand, among GA, birth weight and number of gestations; multi gestations (P=0.0001) was an effective risk factor for IVH occurrence in the controls. Conclusion The incidence of IVH in the IVF group was significantly higher than in the non-IVF group. IVF as an independent risk factor may cause high-grade IVH; however, in the controls, multi gestational pregnancy (P=0.0001) was an effective risk factor for IVH occurrence. © 2019, Iranian Child Neurology Society. All rights reserved

Concomitant Carcinoma in situ in Cystectomy Specimens Is Not Associated with Clinical Outcomes after Surgery

Objective: The aim of this study was to externally validate the prognostic value of concomitant urothelial carcinoma in situ (CIS) in radical cystectomy (RC) specimens using a large international cohort of bladder cancer patients. Methods: The records of 3,973 patients treated with RC and bilateral lymphadenectomy for urothelial carcinoma of the bladder (UCB) at nine centers worldwide were reviewed. Surgical specimens were evaluated by a genitourinary pathologist at each center. Uni- and multivariable Cox regression models addressed time to recurrence and cancer-specific mortality after RC. Results: 1,741 (43.8%) patients had concomitant CIS in their RC specimens. Concomitant CIS was more common in organ-confined UCB and was associated with lymphovascular invasion (p < 0.001). Concomitant CIS was not associated with either disease recurrence or cancer-specific death regardless of pathologic stage. The presence of concomitant CIS did not improve the predictive accuracy of standard predictors for either disease recurrence or cancer-specific death in any of the subgroups. Conclusions: We could not confirm the prognostic value of concomitant CIS in RC specimens. This, together with the discrepancy between pathologists in determining the presence of concomitant CIS at the morphologic level, limits the clinical utility of concomitant CIS in RC specimens for clinical decision-making. Copyright (C) 2011 S. Karger AG, Base