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Enzyme-linked immunosorbent assays (ELISAs) are a cornerstone of modern molecular detection, but the technique still faces notable challenges. One of the biggest problems is discriminating true signal generated by target molecules versus non-specific background. Here, we developed a Si ngle- M olecule C olocalization A ssay (SiMCA) that overcomes this problem by employing total internal reflection fluorescence microscopy to quantify target proteins based on the colocalization of fluorescent signal from orthogonally labeled capture and detection antibodies. By specifically counting colocalized signals, we can eliminate the effects of background produced by non-specific binding of detection antibodies. Using TNF-α, we show that SiMCA achieves a three-fold lower limit of detection compared to conventional single-color assays and exhibits consistent performance for assays performed in complex specimens such as serum and blood. Our results help define the pernicious effects of non-specific background in immunoassays and demonstrate the diagnostic gains that can be achieved by eliminating those effects. A major challenge of enzyme-linked immunosorbent assays is discriminating true signal from non-specific binding. Here the authors present a Single-Molecule Colocalization Assay (SiMCA) which eliminates such effects, enabling reproducible detection of picomolar protein concentrations.

Objective. The aim of this study was to compare National Disability Insurance Scheme (NDIS) timeframes and functional outcomes for a patient population managed in an inpatient hospital rehabilitation unit. Methods. A retrospective hospital audit was undertaken of adult patients admitted to a tertiary-level, regional inpatient rehabilitation unit between January 2017 and December 2021 who were either referred, or not, to the NDIS. A hospital NDIS patient database, Australasian Rehabilitation Outcome Centre episode data, and patient medical records were analysed. The main outcome measures included actual rehabilitation length of stay versus expected length of stay, and Functional Independence Measure (FIM) efficiency for all inpatients, with NDIS timeframes analysed for the NDlS-referred patient subgroup. Results. Rehabilitation inpatients referred for NDIS services significantly exceeded expected rehabilitation length of stay compared to those not referred to the NDIS. Furthermore, expected length of stay was significantly exceeded for those patients who required implementation of a NDIS plan to safely transition from hospital. FIM efficiency was significantly lower for patients referred to the NDIS. Recent improvement in timeframes for being accepted as a NDIS participant did not reduce length of stay. Conclusions. NDIS timeframes for rehabilitation inpatients incur a significant opportunity cost for the provision of efficient inpatient rehabilitation services that are unaccounted for in current benchmarking performance standards.

Background High food insecurity has been demonstrated in rural American Indian households, but little is known about American Indian families in urban settings or the association of food insecurity with diet for these families. The purpose of this study was to examine the prevalence of food insecurity in American Indian households by urban-rural status, correlates of food insecurity in these households, and the relationship between food insecurity and diet in these households. Methods Dyads consisting of an adult caregiver and a child (2–5 years old) from the same household in five urban and rural American Indian communities were included. Demographic information was collected, and food insecurity was assessed using two validated items from the USDA Household Food Security Survey. Factors associated with food insecurity were examined using logistic regression. Child and adult diets were assessed using food screeners. Coping strategies were assessed through focus group discussions. These cross-sectional baseline data were collected from 2/2013 through 4/2015 for the Healthy Children, Strong Families 2 randomized controlled trial of a healthy lifestyles intervention for American Indian families. Results A high prevalence of food insecurity was determined (61%) and was associated with American Indian ethnicity, lower educational level, single adult households, WIC participation, and urban settings ( p  = 0.05). Food insecure adults had significantly lower intake of vegetables ( p  < 0.05) and higher intakes of fruit juice (<0.001), other sugar-sweetened beverages ( p  < 0.05), and fried potatoes ( p  < 0.001) than food secure adults. Food insecure children had significantly higher intakes of fried potatoes ( p  < 0.05), soda ( p  = 0.01), and sports drinks ( p  < 0.05). Focus group participants indicated different strategies were used by urban and rural households to address food insecurity. Conclusions The prevalence of food insecurity in American Indian households in our sample is extremely high, and geographic designation may be an important contributing factor. Moreover, food insecurity had a significant negative influence on dietary intake for families. Understanding strategies employed by households may help inform future interventions to address food insecurity. Trial registration ( NCT01776255 ). Registered: January 16, 2013. Date of enrollment: February 6, 2013.

Patients with pancreas cancer may undergo palliative gastrointestinal or biliary bypass. Recent comparisons of post-operative outcomes following such procedures are lacking. We analyzed patients undergoing exploration, gastrojejunostomy, biliary bypass or double bypass for pancreatic cancer using data from the 2005–2019 American College of Surgeons National Surgical Quality Improvement Program. We compared 30-day mortality and complications across procedures and over time periods (2005-10, 2011-14, 2015-19) using multivariable regression models. Factors associated with postoperative mortality were identified. Of 43,525 patients undergoing surgery with a postoperative diagnosis of pancreatic cancer, 5572 met inclusion criteria. Palliative operations included 1037 gastrojejunostomies, 792 biliary bypasses, 650 double bypasses, and 3093 explorations. The proportion of biliary and double bypass procedures decreased from 2005-10 to 2015-19. Gastrojejunostomy had higher 30-day mortality rate (11.5%) than other operations (p < 0.001). Adjusted 30-day mortality rates remained stable over time (7.8% vs 6.3%, p = 0.095), while rates of serious complications decreased over time (23.2% vs 17.1%, p < 0.001). Palliative bypass for pancreatic cancer has not become safer over time, and 30-day mortality and complications remain high. [Display omitted] •Short-term mortality from palliative operations for pancreatic cancer remains high.•Palliative gastrojejunostomy is associated with the highest 30-day mortality rate of 11.5%.•Rates of major complications from palliative operations have decreased over 15 years.•Rates of double bypass and biliary bypass operations have decreased over time.•Certain risk factors predict high risk of morbidity and short term mortality.

Background Tracheal papillomatosis is a relatively rare condition with limited data on successful treatment modalities. To our knowledge, this is the first report to describe a papilloma arising from an accessory bronchus. Furthermore, this case report demonstrates successful treatment with clinical and patient-centered improvements after use of Spray Cryotherapy. Case presentation A 71-year-old woman presented with one year history of recurrent fevers and intermittent hemoptysis. Imaging and video bronchoscopy revealed an obstructing papilloma of an accessory tracheal bronchus to the right upper lobe. She was treated with debridement followed by multiple cryotherapy treatments resulting in complete clinical and radiographic resolution of her post-obstructive pneumonia. Conclusions This case report not only supports existing literature on the use of cryotherapy for airway diseases but also presents a unique form of obstructing papilloma confined to an accessory bronchus, the only report of its kind based on extensive literature review.

BackgroundHigh-volume centers (HVC), academic centers (AC), and longer travel distances (TD) have been associated with improved outcomes for patients undergoing surgery for pancreatic adenocarcinoma (PAC). Effects of mediating variables on these associations remain undefined. The purpose of this study is to examine the direct effects of hospital volume, facility type, and travel distance on overall survival (OS) in patients undergoing surgery for PAC and characterize the indirect effects of patient-, disease-, and treatment-related mediating variables. Patients and MethodsUsing the National Cancer Database, patients with non-metastatic PAC who underwent resection were stratified by annual hospital volume (< 11, 11–19, and ≥ 20 cases/year), facility type (AC versus non-AC), and TD (≥ 40 versus < 40 miles). Associations with survival were evaluated using multiple regression models. Effects of mediating variables were assessed using mediation analysis. ResultsIn total, 19,636 patients were included. Treatment at HVC or AC was associated with lower risk of death [hazard ratio (HR) 0.90, confidence interval (CI) 0.88–0.92; HR 0.89, CI 0.86–0.91, respectively]. TD did not impact OS. Patient-, disease-, and treatment-related variables explained 25.5% and 41.8% of the survival benefit attained from treatment at HVC and AC, reducing the survival benefit directly attributable to each variable to 4.9% and 6.4%, respectively.ConclusionsTreatment of PAC at HVC and AC was associated with improved OS, but the magnitude of this benefit was less when mediating variables were considered. From a healthcare utilization and cost–resource perspective, further research is needed to identify patients who would benefit most from selective referral to HVC or AC.

BackgroundMinimally invasive distal pancreatectomy (MIDP) is gaining popularity due to improved perioperative outcomes over open distal pancreatectomy (ODP). The purpose of this study is to compare outcomes of MIDP and ODP using patients within a nationwide cohort.MethodsThe American College of Surgeons’ National Quality Improvement Program (2014–2018) was used to evaluate incidence of post-operative pancreatic fistula (POPF) as well as 30-day composite major morbidity for patients undergoing MIDP vs. ODP. Matching was performed with a Mahalanobis-distance model for demographic characteristics, preoperative risk factors, and benign versus malignant pathology. Outcomes were assessed via weighted multiple logistic regression.ResultsA total of 3940 patients underwent distal pancreatectomy (1978 MIDP, 1962 ODP). After matching, 2985 patients were included (1978 MIDP, 1007 ODP). The rates of major morbidity (8.65% MIDP vs. 9.76% ODP, p = 0.37) were similar between groups. The MIDP group was found to have significantly decreased length of stay (5.6 vs. 7 days, p ≤ 0.001), but greater rates (12.54% MIDP vs. 9.35% ODP, p = 0.02) of post-operative fistula.ConclusionsWhen matched for baseline patient characteristics, MIDP was associated with shorter length of hospitalization with similar rates of morbidity compared to ODP. However, MIDP was associated with significantly increased rates of POPF. Further studies are needed to investigate this difference in POPF rate, and determine how to optimize MIDP surgical technique to reduce this risk.

Objective The purpose of this study was to assess the accuracy of BMI categories based on self-reported height and weight in adult women. Methods BMI categories from self-reported responses were compared to categories measured during physical examination from women, age 18 or older, who participated in the National Health and Examination Survey, 1999–2004. We first examined strength of agreement using Cohen’s kappa, which, unlike sensitivity and specificity, allows for the comparison of polychotomous measures beyond chance agreement. Kappa regression identifies potential threats to accuracy. Likelihood of bias, as measured by under-reporting, was examined using logistic regression. Results Cohen’s kappa estimates were 0.443 for pregnant women ( N  = 724) and 0.705 for non-pregnant women ( N  = 5,910). Kappa varied by age and race, but was largely unrelated to socioeconomic status, health and health behaviors. Women who visited a physician in the last year or been diagnosed with osteoporosis were more accurate, while women most likely to under-report were older, white, non-Hispanic, and college-educated. Conclusions Our results suggest substantial agreement between self-reported and measured categories, except for women who are pregnant, above the age of 75 or without physician visits. Under-reporting may be more prevalent in well-educated, white populations than minority populations.

Purpose Despite curative-intent treatment, recurrence is common for patients with colorectal cancer (CRC).  Currently, prediction of disease recurrence and prognostication following surgery is based upon vague clinical factors and more precise and dynamic biomarkers for risk stratification and treatment decisions are urgently needed.  Circulating tumor DNA (ctDNA) is a promising biomarker for patients undergoing treatment for resectable CRC. Methods In this review, we provide an overview of the data supporting current uses of ctDNA for CRC, including localized CRC and resectable colorectal liver metastases (CLM), as well as descriptions of important ongoing clinical trials using ctDNA in the care of patients with CRC. Results The detection of ctDNA following curative-intent therapy is associated with disease recurrence, and multiple trials are investigating its role in determining need and duration for adjuvant therapy for localized CRC. In addition, ctDNA reliably predicts prognosis for patients with CLM, with trials underway studying ctDNA-guided treatment sequencing and intensity. Conclusion The detection of ctDNA is a sensitive and dynamic biomarker for disease recurrence in CRC. Many investigations are underway into ctDNA’s potential role in surveillance and treatment algorithms, and it has the potential to become a critical biomarker to determine individualized strategies for treatment sequencing, choice, and duration of therapies.