Identification of Immunodominant Outer Membrane Proteins of Fusobacterium necrophorum from Severe Ovine Footrot By MALDI-TOF Mass Spectrometry.

The aim of this study was to identify the immunodominant outer membrane proteins (OMPs) of Fusobacterium necrophorum from sheep affected with severe foot-rot. The OMP profile of ovine strains of F. necrophorum has not been well studied. We analyzed the OMP profile of the most frequent lktA variant JKS-F3 of F. necrophorum associated with severe ovine foot-rot with lesion score 4 in order to identify its major immunodominant OMPs. Electrophoretic separations of extracted OMPs showed a number of spots in two-dimensional electrophoretic gels. Two immunoreactive proteins of size around 43 kDa were identified through western blotting using hyperimmune sera raised in rabbits. These two immunogenic OMPs were analyzed by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF/MS) which revealed that these two OMPs of lktA variant JKS-F3 of F. necrophorum showed 46 and 42 percent protein sequence coverage and scores of 125 and 114, respectively, with the reported 43 kDa outer membrane protein of F. necrophorum strain H05, a putative porin having properties similar to pore-forming proteins of anaerobic Gram-negative bacteria. These identified immunogenic OMPs will contribute to our understanding of the pathogenic role played by this organism in ovine foot-rot and could be exploited to devise an effective control strategy through development of an OMP-based recombinant vaccine to mitigate foot-rot in sheep and goats.

Production of Biodegradable Polymer from Agro-Wastes in Alcaligenes sp. and Pseudomonas sp.

The present study was aimed to evaluate the suitability of agro-wastes and crude vegetable oils for the cost-effective production of poly-ß-hydroxybutyrate (PHB), to evaluate growth kinetics and PHB production in Alcaligenes faecalis RZS4 and Pseudomonas sp. RZS1 with these carbon substrates and to study the biodegradation of PHB accumulated by these cultures. Alcaligenes faecalis RZS4 and Pseudomonas sp. RZS1 accumulates higher amounts of PHB corn (79.90% of dry cell mass) and rice straw (66.22% of dry cell mass) medium respectively. The kinetic model suggests that the Pseudomonas sp. RZS1 follows the Monod model more closely than A. faecalis RZS4. Both the cultures degrade their PHB extract under the influence of PHB depolymerase. Corn waste and rice straw appear as the best and cost-effective substrates for the sustainable production of PHB from Alcaligenes faecalis RZS4 and Pseudomonas sp. RZS1. The biopolymer accumulated by these organisms is biodegradable in nature. The agro-wastes and crude vegetable oils are good and low-cost sources of nutrients for the growth and production of PHB and other metabolites. Their use would lower the production cost of PHB and the low-cost production will reduce the sailing price of PHB-based products. This would promote the large-scale commercialization and popularization of PHB as an ecofriendly bioplastic/biopolymer.

Genotyping and antibiotic resistance patterns of Campylobacter fetus subsp.venerealis from cattle farms in India.

Bovine genital campylobacteriosis caused by Campylobacter fetus subsp. venerealis (Cfv) is of considerable economic importance to the cattle industry worldwide. Cfv causes syndrome of temporary infertility in female cattle, early embryonic mortality, aberrant oestrus cycles, delayed conception, abortions and poor calving rates. In the present study, a total of 200 samples obtained from vaginal swabs, cervicovaginal mucous (CVM), preputial washes and semen straws were investigated that were obtained from organized cattle farm of MLRI, Manasbal and unorganized sectors. Out of a total of 200 samples, 49 (47·57%) vaginal swabs, 1 (3·33%) preputial wash and 8 (25%) carried out CVM samples were positive for Cfv, whereas none of the semen straws were positive for Cfv. A total of eleven isolates of Cfv were recovered. PFGE (Pulse field gel electrophoresis) analysis revealed four different pulsotypes (I-IV) circulating in the screened farms. A common pulsotype circulating among farms could not be established. Insertion element (ISCfe1), a 233 bp amplicon of Cfv, was sequenced and the sequence was deposited in GenBank (accession no: MK475662).

Enterotoxin gene profile and molecular epidemiology of Aeromonas species from fish and diverse water sources.

AIMS: This investigation was undertaken to study the prevalence, enterotoxin gene profile and molecular epidemiology of Aeromonads from various sources of water (182) and fish (173). METHODS AND RESULTS: A total of 116 Aeromonas sp. were isolated, of which 48 (26·37%) were from water and 68 (34·62%) were from fish samples collected from retail markets and fish farms. The Aeromonads were recovered from all types of water sources viz. drinking water (13%), surface waters (26%) and fish ponds (69%). The most prevalent species recovered from drinking water was A. hydrophila, from fish ponds it was A. caviae, from surface water sources A. hydrophila and A. caviae were recovered more frequently, and A. hydrophila and A. veronii bv. sobria were isolated predominantly from gills of fish samples. On multiplex PCR analysis for the detection of enterotoxin genes (act, alt, ast), the above mentioned Aeromonas species frequently contained enterotoxin genes, irrespective of their sources. From isolates across all the sources, act (63%) and alt (57%) genes were encountered more frequently than ast (6%). The enterobacterial repetitive intergenic consensus sequences polymerase chain reaction was used for typing of isolates and most of the isolates from water and fish were related, owing to similar ecosystem. CONCLUSION: A wide distribution of enterotoxin genes in Aeromonads from water and fish is a potential public health threat and molecular genotyping can be helpful to study epidemiology of the pathogen. SIGNIFICANCE AND IMPACT OF THE STUDY: A high proportion of isolates recovered from diverse water sources, particularly potable drinking water and fish samples carried one or more enterotoxin genes thereby indicating a potential pathogenic nature of isolates from these sources. The genetic relatedness was detected amongst many isolates recovered from water sources and fish samples indicating circulation of familiar virulent clones in the aquatic environments.

Dysplasia severity is associated with poor quality of life in patients with Barrett's esophagus referred for endoscopic eradication therapy.

Limited data exist regarding patient-reported outcomes and quality of life (QOL) experienced by patients with Barrett's esophagus (BE) referred for endoscopic eradication therapy (EET). Specifically, the impact of grade of dysplasia has not been explored. The purpose of this study is to measure patient-reported symptoms and QOL and identify factors associated with poor QOL in BE patients referred for EET. This was a prospective multicenter study conducted from January 2015 to October 2017, which included patients with BE referred for EET. Participants completed a set of validated questionnaires to measure QOL, symptom severity, and psychosocial factors. The primary outcome was poor QOL defined by a PROMIS score >12. Multivariable logistic regression analysis was performed to identify factors associated with poor QOL. In total, 193 patients participated (mean age 64.6 years, BE length 5.5 cm, 82% males, 92% Caucasians) with poor QOL reported in 104 (53.9%) participants. On univariate analysis, patients with poor QOL had lower use of twice daily proton pump inhibitor use (61.5% vs. 86.5%, P = 0.03), shorter disease duration (4.9 vs. 5.9 years, P = 0.04) and progressive increase in grade of dysplasia (high-grade dysplasia: 68.8% vs. 31.3%, esophageal adenocarcinoma: 75.5% vs. 24.5%, P < 0.001). Multivariate analysis demonstrated that high-grade dysplasia was independently associated with poor QOL (OR: 5.57, 95% CI: 1.05, 29.5, P = 0.04). In summary, poor QOL is experienced by the majority of patients with BE referred for EET and the degree of dysplasia was independently associated with poor QOL, which emphasizes the need to incorporate patient-centered outcomes when studying treatment of BE-related dysplasia.

The detection and prevalence of leukotoxin gene variant strains of Fusobacterium necrophorum in footrot lesions of sheep in Kashmir, India.

The objective of this study was to determine the prevalence and identification of leukotoxin gene, lktA, variant strains of Fusobacterium necrophorum in the footrot lesions of sheep. The detection of F. necrophorum was carried out by PCR targeting the lktA gene fragment and identification of lktA variant strains was done by PCR-single stranded conformational polymorphism (PCR-SSCP) and gene sequencing. Of the 450 swabs collected from footrot lesions of sheep, 117 were lktA-positive for F. necrophorum. Of the 50 swabs collected from apparently asymptomatic sheep, only one was lktA-positive for F. necrophorum. The overall prevalence of F. necrophorum in footrot affected sheep in Kashmir valley was 26%, and ranged from 20 to 34.8%, respectively. PCR-SSCP of lktA gene fragment analysis revealed three lktA variants, designated as JKS-F1/F2/F3, while two samples (1.7%) showed multiple lktA variant strains of F. necrophorum in a single footrot-affected sheep hoof. This appears to be the first report on the presence of more than one lktA variant of F. necrophorum in a footrot lesion of sheep. The JKS-F3 lktA variant was the most frequent (75.4%), followed by JKS-F2 (14.4%) and JKS-F1 (8.4%), respectively. Among the three lktA variants identified, JKS-F3 was detected in 74 (86.0%) samples from severe footrot affected sheep with a lesion score of 4. The data suggest that JKS-F3 is the predominant lktA variant of F. necrophorum and is associated with severe footrot in sheep. Hence, JKS-F3 may be a significant variant contributing to the severity and duration of the disease in sheep.

Does time of surgery and complication have any correlation in the management of hip fracture in elderly and can early surgery affect the outcome?

OBJECTIVE: Hip fractures in the elderly are usually associated with a high rate of morbidity and mortality and affect quality of life. On review of published data, the current guidelines in the literature indicate that early surgery should be performed within 24 h of injury because it is associated with better functional outcomes and lower rates of perioperative complications and mortality. The aim of this study was to evaluate whether any correlation exists between early surgical intervention and functional outcomes. METHODS: In a retrospective study, we analyzed the records between January 2010 and December 2016 and 550 elderly patients between the age group of 65-95 were included. RESULTS: The results of early surgery were promising, 451 (82%) patients did not have any complication, 42 (7.6%) has developed bed sore, 23 (4%) developed a hospital-acquired infection, 13 (2.3%) developed wound infection, 08 (1.4%) developed DVT, 04 (0.7%) developed implant failure, and 09 (01%) died of some or other complications. CONCLUSION: Still, there is conflicting evidence in the published data that early surgery would improve mortality and function. However, early surgery definitively improves outcome and reduces morbidity, bed sores, infection, and the length of hospital stay could be improved by reducing the waiting time of hip surgery following fracture. We concluded that it is beneficial to the elderly patients to receive surgical treatment as early as possible.

Incorporation of carrot pomace powder in wheat flour: effect on flour, dough and cookie characteristics.

Carrot pomace powder (CPP) of 72 and 120 mesh sizes was incorporated in wheat flour at 10, 15 and 20 % level and its impact on flour, dough and cookie characteristics was evaluated. Protein content of the flour blends (8.84-7.88 %) decreased and fibre content (4.63-6.68 %) increased upon blending of CPP in wheat flour. Wheat flour containing 120 mesh CPP showed better functional properties [water absorption (1.16-1.47 %), oil absorption (1.11-1.39 %), solubility index (41-50 %) and swelling power (1.34-1.39)] than those containing 72 mesh. Water solvent retention capacity and sucrose solvent retention capacity increased while lactic acid solvent retention capacity and sodium carbonate solvent retention capacity decreased with blending of CPP. Water absorption, dough development time and degree of softening increased whereas, dough stability and mixing tolerance decreased with increasing CPP. The highest decrease in pasting was observed flour containing 72 mesh CPP. Rheology of dough containing 120 mesh CPP closely resembled the control. Color of flour and cookies increased with blending of CPP irrespective of mesh size. Antioxidant activity of cookies was higher than the flour blends. The cookies containing CPP of 72 mesh showed the lowest hardness. However, cookies containing CPP of 120 mesh showed the best sensory properties. Incorporation of 120 mesh CPP produced low gluten cookies with manageable flour and dough characteristics and better antioxidant and sensory properties.

Ultrasound treatment: effect on physicochemical, microbial and antioxidant properties of cherry (Prunus avium).

The cherry was treated with ultrasonic waves (33 kHz, 60 W) at different time intervals (10, 20, 30, 40, 60 min) and study was carried out to analyze the change in physico-chemical properties (TSS, pH, color, acidity and firmness), antioxidant potential and microbial load of the fruit during the storage period of 15 days at 4 °C. It was observed that ultrasound treatment (US) between 30 and 40 min showed better retention of color of the fruit during the storage period. The antioxidant assays (DPPH, ABTS and TPC) also increased significantly (P ≤ 0.05) up to 40 min, however the firmness of the fruit was affected and it showed a significant decrease beyond 20 min of US treatment. The sample with 40 min US treatment showed significantly less microbial load than other samples. The 20-40 min US treatment time (33 kHz, 60 W) was suggested for preservation of cherry during the storage at 4 °C.

Application of guar-xanthan gum mixture as a partial fat replacer in meat emulsions.

The physicochemical, oxidative, texture and microstructure properties were evaluated for low fat meat emulsions containing varying levels of guar/xanthan gum mixture (1:1 ratio) as a fat substitute. Partial replacement of fat with guar/xanthan gum resulted in higher emulsion stability and cooking yield but lower penetration force. Proximate composition revealed that high fat control had significantly higher fat and lower moisture content due to the difference in basic formulation. Colour evaluation revealed that low fat formulations containing gum mixture had significantly lower lightness and higher yellowness values than high fat control formulation. However non-significant difference was observed in redness values between low fat formulations and the high fat control. The pH values of the low fat formulations containing gum mixture were lower than the control formulations (T0 and TC). The MetMb% of the high fat emulsion formulation was higher than low fat formulations. The significant increase of TBARS value, protein carbonyl groups and loss of protein sulphydryl groups in high fat formulation reflect the more oxidative degradation of lipids and muscle proteins during the preparation of meat emulsion than low fat formulations. The SEM showed a porous matrix in the treatments containing gum mixture. Thus, the guar/xanthan gum mixture improved the physicochemical and oxidative quality of low fat meat emulsions than the control formulations.

Status of Competency-Based Medical Education in Endoscopy Training: A Nationwide Survey of US ACGME-Accredited Gastroenterology Training Programs.

OBJECTIVES: The Accreditation Council for Graduate Medical Education (ACGME) emphasizes the importance of medical trainees meeting specific performance benchmarks and demonstrating readiness for unsupervised practice. The aim of this study was to examine the readiness of Gastroenterology (GI) fellowship programs for competency-based evaluation in endoscopic procedural training. METHODS: ACGME-accredited GI program directors (PDs) and GI trainees nationwide completed an online survey of domains relevant to endoscopy training and competency assessment. Participants were queried about current methods and perceived quality of endoscopy training and assessment of competence. Participants were also queried about factors deemed important in endoscopy competence assessment. Five-point Likert items were analyzed as continuous variables by an independent t-test and χ(2)-test was used for comparison of proportions. RESULTS: Survey response rate was 64% (94/148) for PDs and 47% (546/1,167) for trainees. Twenty-three percent of surveyed PDs reported that they do not have a formal endoscopy curriculum. PDs placed less importance (1­very important to 5­very unimportant) on endoscopy volume (1.57 vs. 1.18, P<0.001), adenoma detection rate (2.00 vs. 1.53, P<0.001), and withdrawal times (1.96 vs. 1.68, P=0.009) in determining endoscopy competence compared with trainees. A majority of PDs report that competence is assessed by procedure volume (85%) and teaching attending evaluations (96%). Only a minority of programs use skills assessment tools (30%) or specific quality metrics (28%). Specific competencies are mostly assessed by individual teaching attending feedback as opposed to official documentation or feedback from a PD. PDs rate the overall quality of their endoscopy training and assessment of competence as better than overall ratings by trainees. CONCLUSIONS: Although the majority of PDs and trainees nationwide believe that measuring specific metrics is important in determining endoscopy competence, most programs still rely on procedure volume and subjective attending evaluations to determine overall competence. As medical training transitions from an apprenticeship model to competency-based education, there is a need for improved endoscopy curricula which are better suited to demonstrate readiness for unsupervised practice.

Xanthan gum as a fat replacer in goshtaba-a traditional meat product of India: effects on quality and oxidative stability.

Goshtaba is a restructured meat product of Kashmiri wazwan prepared from meat emulsion with added fat (20 %), salt, spices and condiments and cooked in the curd. The present study was undertaken for the development of low fat goshtaba with the addition of xanthan gum as a fat replacer and was evaluated for proximate composition, pH, colour, lipid and protein oxidation, texture, microstructure and sensory properties. Low fat goshtaba formulations containing xanthan gum were higher in protein and moisture contents but, lower in fat content and pH value than the high fat control (p < 0.05). Colour evaluation revealed that high fat goshtaba had significantly higher L* value, but lower a* value than its low fat counterparts (p < 0.05). The significant decrease of TBARS values, protein carbonyls and loss of protein sulphydryl groups in low fat goshtaba formulations reflects the potential antioxidant activity of xanthan gum (p < 0.05). Hardness was significantly higher in high fat control but, cohesiveness, gumminess, and chewiness did not show any significant difference. Springiness increased with the increasing concentration of xanthan gum (0.5-1.5 %) and was higher in low fat product containing 1.5 % xanthan gum. SEM results indicate that xanthan gum lead to formation of an additional gel network which holds more water. Sensory evaluation revealed that goshtaba product with 0.5 % xanthan gum had quality characteristics that were similar to the control product containing 20 % fat.

Characterization of cookies made from wheat flour blended with buckwheat flour and effect on antioxidant properties.

Buckwheat flour was incorporated into wheat flour at different levels (0, 20, 40, 60, 80, and 100 %) and the physicochemical, functional and antioxidant properties of the blended flour were studied. This study also investigated the effect of buckwheat on the retention of antioxidant properties of cookies during baking. The results showed significant variation in physicochemical and functional properties of the blended flour. The addition of buckwheat flour into wheat flour also increased the antioxidant properties of blended flour proportionally, but metal chelating properties decreased. The incorporation of buckwheat in wheat flour helped in better retention of antioxidant potential of cookies during baking process as buckwheat cookies (100 % buckwheat) showed greater percentage increase in antioxidant properties than control (100 % wheat). Quality characteristics of cookies such as hardness and spread ratio decreased, while as non-enzymatic browning (NEB) increased significantly with increase in the proportion of buckwheat flour in wheat flour. The Overall acceptability of cookies by sensory analysis was highest at 40 % level of blending. This study concluded that addition of buckwheat in wheat flour, may not only improve the physico-chemical and functional properties of the blended flour but may also enhance the nutraceutical potential of the product prepared from it.

Effect of green tea powder on thermal, rheological & functional properties of wheat flour and physical, nutraceutical & sensory analysis of cookies.

Green tea powder (GTP) was incorporated in wheat flour at different levels (1, 2 and 4 %) and its effect on quality parameters of flour and cookies were studied. Thermal analysis showed that addition of GTP did not increase the onset temperature, end temperature and enthalpy of flour significantly. However, the increase in GTP content retarded the retro gradation of the gelatinized wheat flour gel and decreased the water absorption capacity, Oil absorption capacity, water solubility index, Foam capacity and foam stability significantly. The L* and b* values of cookies decreased from 53.63 to 33.23 and 29.02 to 24.39 respectively, while as, a* value showed an increase from 3.61 to 5.23 indirect proportion to quantity added. The addition of GTP to wheat flour increased the DPPH (2,2-diphenyl-1-picrylhydrazyl) percent inhibition and reducing power of cookies. Further, it also increases the acceptability for color, aroma and taste; however it loosened its structural integrity and decreased the fracture force.

A multiplex PCR for detection of enterotoxin genes in Aeromonas species isolated from foods of animal origin and human diarrhoeal samples.

AIMS: The present study describes incidence and enterotoxin gene profile of Aeromonas spp. from human diarrhoeal samples (83) and raw meats (171). METHODS AND RESULTS: The samples were screened for isolation of Aeromonads. Aeromonas spp. contaminated raw meats of all kinds under the study and per cent contamination in chicken, mutton and beef was 14·03, 22·89 and 19·35, respectively. Of the 83 diarrhoeal samples from children, 6 (7·22%) were positive for presence of Aeromonas spp. Seven different species of Aeromonas (Aer. hydrophila, Aer. caviae, Aer. veronii bv sobria, Aer. trota, Aer. schubertii, Aer. jandaei and Aer. allosaccharophila) could be identified from foods and from diarrhoeal samples two species (Aer. caviae and Aer. hydrophila) were encountered. Unique primers were designed, and a multiplex PCR was standardized for detection of three enterotoxin genes (act, alt, ast) in the Aeromonas spp. Of the 39 isolates, 35 (89·74%) carried one or more enterotoxin genes: act, alt and ast genes were detected in 30 (76·92%), 31 (79·48%) and 4 (10·25%) isolates, respectively. The enterotoxin genes from a strain recovered from mutton were sequenced and submitted to GenBank and the accession no.s KC687135, KC633828 and KC687134 were provided for alt, ast and act, respectively, by the GenBank. CONCLUSIONS: The occurrence of enterotoxigenic Aeromonads in raw meats and diarrhoeal samples is a public health concern. SIGNIFICANCE AND IMPACT OF THE STUDY: Given the increasing evidence of involvement of Aeromonads in foodborne outbreaks, the standardization of single-step multiplex PCR will be helpful tool for detection of enterotoxin genes in Aeromonas spp.

Presence or absence of intestinal metaplasia but not its burden is associated with prevalent high-grade dysplasia and cancer in Barrett's esophagus.

Universal agreement on the inclusion of intestinal metaplasia to diagnose Barrett's esophagus (BE) is lacking. Our aim was to determine the association of intestinal metaplasia and its density with the prevalence of dysplasia/cancer in columnar lined esophagus (CLE). Patients with CLE but no intestinal metaplasia (CLE-no IM) were identified by querying the clinical pathology database using SNOMED codes for distal esophageal biopsies. CLE-IM patients were identified from a prospectively maintained database of BE patients. Subsequently, relative risks for prevalent dysplasia and cancer were calculated. Since patients with CLE-no IM are not usually enrolled in surveillance, only prevalent dysplasia/cancer on index endoscopy was analyzed. Goblet cell density and percent intestinal metaplasia were estimated. All biopsy slides were reviewed for dysplasia by two experienced gastrointestinal pathologists. Two hundred sixty-two CLE-IM and 260 CLE-no IM patients were included (age 64±12 vs. 60±11 years, P=0.001; whites 92% vs. 82%, P=0.001; males 99.7% vs. 99.3%, P=NS; CLE length 3.4±3.2 vears 1.4±0.4 cm, P=0.001 and hiatus hernia 64% vs. 56%, P=0.013). The odds of finding low-grade dysplasia and of high-grade dysplasia (HGD)/cancer were 12.5-fold (2.9-53.8, P=0.007) and 4.2-fold (95% CI 1.4-13, P=0.01) higher, respectively, in the CLE-IM group. Reanalysis after controlling for important variables of age, race, and length did not significantly alter the overall results. In CLE-IM group, when patients with high (>50/LPF) versus low goblet cell density (<50/LPF) and <10% versus >10% intestinal metaplasia were compared, the odds of HGD/cancer, OR 1.5 (0.5-4.9, P=0.5) and 1.97 (0.54-7.22), respectively, were not significantly higher. Demonstration of intestinal metaplasia continues to be an essential element in the definition of BE, but its quantification may not be useful for risk stratification of HGD/cancer in BE.

Evaluation of the updated confocal laser endomicroscopy criteria for Barrett's esophagus among gastrointestinal pathologists.

Previously developed novel probe-based confocal laser endomicroscopy (pCLE) criteria have been found to have high accuracy and substantial interobserver agreement (IOA) for diagnosing dysplasia in Barrett's esophagus (BE) when used by endoscopists. These updated criteria are: (i) epithelial surface: saw toothed, (ii) cells: enlarged, (iii) cells: pleomorphic, (iv) glands: not equidistant, (v) glands: unequal in size and shape, and (vi) goblet cells: not easily identified. The accuracy and IOA among pathologists in the diagnosis of dysplasia using the novel pCLE criteria is not known. The primary objective of the study was to evaluate the accuracy, overall IOA and learning curve among three gastrointestinal (GI) pathologists in diagnosing dysplasia in BE using the updated pCLE criteria. The secondary aim was to compare the accuracy and IOA between GI pathologists and gastroenterology endoscopists. Ninety pCLE videos and respective histology were retrieved from a previously conducted multicenter, prospective, randomized, controlled trial evaluating the utility of pCLE in BE patients. Videos were obtained from 101 BE patients previously enrolled for surveillance or endoscopic treatment of high-grade dysplasia or early esophageal adenocarcinoma. Three GI pathologists reviewed 90 pCLE video clips for dysplasia versus no dysplasia, confidence in their diagnosis, and image quality. The overall accuracy for the diagnosis of dysplasia (low-grade dysplasia/high-grade dysplasia/esophageal adenocarcinoma) was 77.8% (95% confidence interval [CI]: 72.4-82.3). The accuracy was higher when pathologists had 'high confidence' in their assessment of the videos (93.8% vs. 69.3%, P < 0.001). There was no significant difference in accuracy between the first set of 30 and second set of 60 videos (84% vs. 74%, P = 0.065). IOA among GI pathologists was substantial, k = 0.65 (95% CI: 0.53-0.73). The sensitivity for detecting dysplasia was 85% (95% CI: 78.1-90.7) and the specificity was 70% (95% CI: 61.91-77.92). These results were comparable with the evaluation of the same set of videos by endoscopists. GI pathologists have high accuracy and substantial IOA for diagnosing BE dysplasia with pCLE. Pathologists appear to have similar accuracy and IOA as endoscopists. These results provide further support of endoscopists accurately interpreting the in vivo optical histology provided by pCLE.

Tissue informative cell-free DNA methylation sites in amyotrophic lateral sclerosis.

Cell-free DNA (cfDNA) is increasingly recognized as a promising biomarker candidate for disease monitoring. However, its utility in neurodegenerative diseases, like amyotrophic lateral sclerosis (ALS), remains underexplored. Existing biomarker discovery approaches are tailored to a specific disease context or are too expensive to be clinically practical. Here, we address these challenges through a new approach combining advances in molecular and computational technologies. First, we develop statistical tools to select tissue-informative DNA methylation sites relevant to a disease process of interest. We then employ a capture protocol to select these sites and perform targeted methylation sequencing. Multi-modal information about the DNA methylation patterns are then utilized in machine learning algorithms trained to predict disease status and disease progression. We applied our method to two independent cohorts of ALS patients and controls (n=192). Overall, we found that the targeted sites accurately predicted ALS status and replicated between cohorts. Additionally, we identified epigenetic features associated with ALS phenotypes, including disease severity. These findings highlight the potential of cfDNA as a non-invasive biomarker for ALS.

The diagnostic accuracy of 22-gauge and 25-gauge needles in endoscopic ultrasound-guided fine needle aspiration of solid pancreatic lesions: a meta-analysis.

BACKGROUND AND STUDY AIMS: It is uncertain if needle gauge impacts the diagnostic accuracy of endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) of pancreatic mass lesions. Our aim was to use meta-analysis to more robustly define the diagnostic accuracy of EUS-FNA for pancreatic masses using 22 G and 25 G needles. PATIENTS AND METHODS: Studies were identified by searching nine medical databases for reports published between 1994 and 2011, using a reproducible search strategy comprised of relevant terms. Only studies comparing the overall diagnostic accuracy of 22 G vs. 25 G EUS needles that used surgical histology or at least 6 months clinical follow up for a gold standard were included. Two reviewers independently scored the identified studies for methodology and abstracted pertinent data. When required, the original investigators were contacted to provide additional data. Pooling was conducted by both fixed-effects and random-effects models. Diagnostic characteristics (sensitivity, specificity, positive and negative likelihood ratios) with 95% confidence intervals (CIs) were calculated. RESULTS: Eight studies involving 1292 subjects met the defined inclusion criteria. Of the 1292 patients, 799 were in the 22 G group and 565 were in the 25 G group (both needles were used in 72 patients). The pooled sensitivity and specificity of the 22 G needle were 0.85 (95%CI 0.82-0.88) and 1 (95%CI 0.98-1) respectively. The pooled sensitivity and specificity of the 25 G needle were 0.93 (95%CI 0.91-0.96) and 0.97 (95%CI 0.93-0.99) respectively. The bivariate generalized linear random-effect model indicated that the 25 G needle is associated with a higher sensitivity (P = 0.0003) but comparable specificity (P = 0.97) to the 22 G needle. CONCLUSIONS: This meta-analysis suggests 25 G needle systems are more sensitive than 22 G needles for diagnosing pancreatic malignancy.

Effect of acid-suppressive therapy on narrow band imaging findings in gastroesophageal reflux disease: a pilot study.

Standard endoscopy is an insensitive test for gastroesophageal reflux disease (GERD). Narrow band imaging (NBI) endoscopy enhances visualization of the distal esophagus. NBI patterns like intrapapillary capillary loop (IPCL) dilatation, tortuosity, and increased number; microerosions; increased vascularity at the squamocolumnar junction (SCJ); ridge-villous pattern below the SCJ; and presence of columnar islands in the distal esophagus have been suggested as features of GERD. We evaluated the effect of proton pump inhibitor (PPI) therapy on NBI findings in GERD patients. Patients prospectively underwent NBI upper endoscopy before and after PPI therapy. NBI findings were recorded at each endoscopy. Twenty-one patients with GERD symptoms (mean age 60.0 years; males 90.5%; white 90.5%) were studied. After PPI therapy, there was a significant reduction in the proportion of patients with the following NBI features: IPCL tortuosity (90% vs. 4.8%, P < 0.0001), dilated IPCLs (86% vs. 9.5%, P < 0.0001), and increased vascularity at the SCJ (43% vs. 9.5%, P= 0.0082). PPI led to healing of all microerosions (71% vs. 0%, P < 0.0001) and disappearance of ridge-villous patterns below the SCJ (14% vs. 0%, P < 0.0001). There was no significant change in the proportion of patients with increased numbers of IPCLs pre- and post-PPI therapy (71% vs. 48%, P= 0.09) or columnar islands in the distal esophagus (38% vs. 29%, P= 0.31). In patients with GERD symptoms, NBI features suggestive of GERD respond to PPI; suggesting these features are truly acid-mediated. These findings need to be confirmed by randomized controlled trials.