Development of an Automated Capillary Immunoassay to Detect Prion Glycotypes in Creutzfeldt-Jakob Disease.

Creutzfeldt-Jakob disease (CJD) comprises a group of transmissible neurodegenerative diseases with vast phenotypic diversity. Sporadic CJD heterogeneity is predominantly influenced by the genotype at codon 129 of the prion-encoding gene and the molecular weight of PrPSc fragments after protease digestion, resulting in a classification of 6 subtypes of CJD (MM1, MM2, MV1, MV2, VV1, and VV2). The majority of cases with CJD can be distinguished using this classification system. However, a number of reported CJD cases are phenotypically unique from others within their same subtype, such as variably protease-sensitive prionopathies, or exist as a mixture of subtypes within the same patient. Western blotting of brain tissue, along with the genotyping of codon 129 of the prion-encoding gene, is considered the "gold standard" for the biochemical characterization of CJD. Western blotting requires a significant amount of prion protein for detection, is labor-intensive, and is also associated with high interassay variability. In addition to these limitations, a growing body of research suggests that unique subtypes of CJD are often undetected or misdiagnosed using standard diagnostic western blotting protocols. Consequently, we successfully optimized and developed a capillary-based western assay using the JESS Simple Western (ProteinSimple) to detect and characterize prion proteins from patients with CJD. We found that this novel assay consistently differentiated CJD type 1 and type 2 cases with a limit of detection 10 to 100× higher than traditional western blotting. Cases with CJD in which type 1 and type 2 coexist within the same brain region can be detected using type 1-specific and type 2-specific antibodies, and we found that there was remarkable specificity for the detection of cases with variably protease-sensitive prionopathy. The assay presented displays outstanding sensitivity, allowing for the preservation of valuable samples and enhancing current detection methods.

Filling the gaps in knowledge translation: Physical therapy recommendations for individuals with spinal muscular atrophy compared to standard of care guidelines.

This study describes the current landscape of physical therapy practice recommendations in the United States for children with spinal muscular atrophy (SMA) and their relationship to current SMA standard of care (SMA-SOC) guidelines. Pediatric physical therapists were surveyed to determine their knowledge of SMA-SOC guidelines, and the type, duration and frequency of intervention they recommend for children with SMA, as well as perceived barriers and facilitators to progress in physical therapy. Physical therapists recommend five key intervention areas for individuals with SMA; however discrepancies exist between the SOC recommended intervention parameters and respondents' reported frequency and duration of recommendations. After individuals with SMA initiated disease modifying pharmacotherapies, a majority of physical therapist respondents recommended increases in both frequency and duration of interventions. Nearly all respondents reported that familiarity with SOC guidelines was beneficial to their practice. The primary facilitator to progress was parent/caregiver support, while the primary barrier was limited access to resources. Variation in practice exists regarding care for those with SMA, particularly in the areas of frequency and duration of specific interventions. These findings can guide educational initiatives, identify future research needs and further inform SMA-SOC and best-practice rehabilitation management.

Prospective Study Demonstrates Utility of EP-QuIC in Creutzfeldt-Jakob Disease Diagnoses.

Prospectively acquired Canadian cerebrospinal fluid samples were used to assess the performance characteristics of three ante-mortem tests commonly used to support diagnoses of Creutzfeldt-Jakob disease. The utility of the end-point quaking-induced conversion assay as a test for Creutzfeldt-Jakob disease diagnoses was compared to that of immunoassays designed to detect increased amounts of the surrogate markers 14-3-3γ and hTau. The positive predictive values of the end-point quaking-induced conversion, 14-3-3γ, and hTau tests conducted at the Prion Diseases Section of the Public Health Agency of Canada were 96%, 68%, and 66%, respectively.

Creutzfeldt-Jakob Disease with a Five-Year Clinical Course, Multicentric Cerebellar Prion Plaques and Prior History of Biopsy-Proven Primary Angiitis of the Central Nervous System: A Case for Iatrogenic Exposure?

Creutzfeldt-Jakob disease (CJD) is a rapidly progressive neurodegenerative disease that can arise spontaneously, genetically, or be acquired through iatrogenic exposure. Most patients die within a year of symptom onset. It is rare, affecting 1-2 per million per year, and the majority of cases are sporadic. Primary angiitis of the central nervous system (PACNS) is also rare, affecting 2.4 per million per year. We present a case of an unusually long clinical course of CJD, almost five years, which began with symptoms of apraxia. The patient had biopsy-proven PACNS 16 years prior to clinical presentation, and the site of biopsy was the left parietal lobe. Autopsy revealed multicentric prion plaques in the cerebellum, in the setting of normal genetic testing. The presence of plaques in the cerebellum, and prior neurosurgery, raises the possibility of iatrogenic exposure. We present the details of this case, including pathology from the original biopsy and final autopsy, as well as a review of relevant cases in the literature.

Cannabis in homes with children: use and storage practices in a legalised state.

Our objective was to examine the prevalence of cannabis in homes with children and temporal trends in storage practices among caregivers. Caregivers of children ages 1-14 in Colorado, identified through a representative population-based telephone survey, participated in a repeated telephone survey from 2014 to 2017. Representing 79 805 households in Colorado with children, 8.6% (95% CI 7.3 to 10.0) of caregivers reported cannabis in the home. From 2014 to 2017, the prevalence of reporting cannabis in the home increased significantly from 6.9% to 11.2% (p=0.02). Among caregivers who reported cannabis in the home, 91.4% reported storage locations inaccessible to children, such as 'out of reach'. Fewer, 67.0%, caregivers reported storage in a locked container, such as a cabinet, drawer or safe. In a state with recreational cannabis, an increasing number of children live in homes where cannabis is present. Public health practitioners should promote messages to caregivers about safe cannabis storage practices.

Impact of an incentive-based prenatal smoking cessation program for low-income women in Colorado.

OBJECTIVE: To assess birth outcomes and cost-savings of an incentive-based prenatal smoking cessation program targeting low-income women in Colorado. DESIGN: Prospective observational cohort with nonequivalent population control groups. SAMPLE: Program participants (n = 2,231) linked to the birth certificate to ascertain birth outcomes compared to two reference populations from Pregnancy Risk Assessment Monitoring System (PRAMS) and Colorado live births based on the birth certificate. MEASUREMENTS: Tobacco cessation metrics in the third trimester of pregnancy, neonatal low birth weight (<2,500 g), preterm birth (birth at <37 weeks gestation), neonatal intensive care unit (NICU) admission and maternal gestational hypertension. Cost-savings and return on investment (ROI) were projected using average Medicaid reimbursement. RESULTS: Infants of mothers enrolled in the program had a lower risk of low birthweight (RR = 0.86; 95% CI = 0.75, 0.97), preterm birth (PTB) (RR = 0.76; 95% CI = 0.65, 0.88) and neonatal intensive care unit (NICU) admission (RR = 0.76; 95% CI = 0.66, 0.88) compared to the birth certificate population, corresponding to a ROI of $7.73 and an individual cost savings of $6,040. Compared to PRAMS, infants of enrolled mothers had a lower risk of PTB (RR = 0.72; 95% CI = 0.53, 0.99) and NICU admission (RR = 0.45; 95% CI = 0.32, 0.62), corresponding to an ROI of $2.79 and an individual cost savings of $2,182. CONCLUSIONS: We found a reduction of adverse birth outcomes, and cost savings.

Investigating the Aftershock of a Disaster: A Study of Health Service Utilization and Mental Health Symptoms in Post-Earthquake Nepal.

Background: In 2015, a 7.8 magnitude earthquake struck Nepal, causing unprecedented damage and loss in the mountain and hill regions of central Nepal. The aim of this study was to investigate the association between healthcare access and utilization, and post-disaster mental health symptoms. Methods: A cross-sectional study conducted with 750 disaster-affected individuals in six districts in central Nepal 15 months post-earthquake. Anxiety and depression were measured through the Depression, Anxiety and Stress Scale (DASS-21). Healthcare utilization questions examined types of healthcare in the communities, utilization, and approachability of care providers. Univariate analyses, ANOVAs and Tobit regression were used. Results: Depression and anxiety symptoms were significantly higher for females and individuals between 40-50 years old. Those who utilized a district hospital had the lowest anxiety and depression scores. Participants who indicated medical shops were the most important source of health-related information had more anxiety and depression than those who used other services. Higher quality of healthcare was significantly associated with fewer anxiety and depressive symptoms. Conclusions: Mental health symptoms can last long after a disaster occurs. Access to quality mental health care in the primary health care settings is critical to help individuals and communities recover immediately and during the long-term recovery.

Translating medicines to patients: A novel methodology for quantifying the global medical supplies and donations program.

BACKGROUND: International medical donation programs can help alleviate the burden of illness and serve as a safety net for the global health care system. However, to our knowledge no studies have assessed the number of individuals served through medical donation programs. As such, this study aimed to evaluate the impact of the Americares Foundation's (Americares) medical donation program in terms of the number of patients served. METHODS: We conducted an outcome evaluation study in 34 health facilities in 10 countries that receive medical donations from Americares. Medical records were randomly sampled at each participating facility and evaluated for types of medications and number of courses of prescribed treatments. Facility level data and donation inventory data were also collected. We developed an algorithm for converting quantities of donated medicines into the number of individuals served at the facility level. These estimates were then extrapolated to the country and region levels to assess the total impact of medications donated in 2015. Probabilistic sensitivity analysis was conducted to derive 95% credible ranges for projected estimates and to assess model uncertainty. RESULTS: Records of 3,205 unique patients were reviewed, encompassing 10,449 medical visits. The average number of medications and courses of treatments prescribed per visit were 2.63 and 2.68, respectively. The average medication destruction rate ranged from 0% to 24% at facilities, with a cross-country average of 7%. For the 10 countries included in the study, we project that 700,377 unique individuals were served through the program (95% credible range: 518,401-905,982). Scaled across all regions receiving Americares donations, we project that the program supported an estimated 5.1 million beneficiaries, including 484,188 chronic care and 4.65 million acute care patients. CONCLUSIONS: This study provides a novel methodology for medical donation programs seeking to estimate one of their key outcomes-patients served-and global reach. Rigorous assessments of program outcomes can provide important insights into the value of medical donation initiatives. TRIAL REGISTRATION: Human subjects approval was received from the University of Washington Institutional Review Board (Approval #52316; 7/19/2016).

Prenatal screening for substance use and violence: findings from physician focus groups.

OBJECTIVE: To guide quality improvement activities, the study sought to identify effective strategies for influencing and improving physician screening and referral of pregnant women for violence and substance abuse (alcohol, drugs and tobacco). METHODS: This qualitative study conducted in Washington State consisted of interviews with eight physicians and focus groups with twenty-eight physicians who practice obstetric care. Physicians, selected using systematic sampling, were asked about perceptions on the importance of screening and barriers to effective screening, awareness of information and resources from the state Department of Health, and the effectiveness of various provider training strategies for improving prenatal screening. RESULTS: Physicians were most interested in practical, concise information for themselves and office staff. Referral information and patient handouts were identified as important tools to increase the efficacy of screening and intervention. Physicians supported in-person programs in offices or in hospitals but rejected use of audio conferences and direct mailings. CONCLUSIONS: This study provided insight about the way we deliver best practice information to physicians. Collecting qualitative data from physicians is important prior to developing statewide quality improvement activities aimed at this group.

Decreased caspase-3 activity in human lens epithelium from posterior subcapsular cataracts.

Apoptosis has been implied in normal lens development in the embryo as well as in lens fibre differentiation. It has also been suggested to play a role in non-congenital cataract and in the formation of posterior subcapsular opacification, but data on the presence of apoptosis in human lens epithelium from cataractous lenses are scarce and conflicting. The present study aimed to investigate apoptosis in lens epithelium from patients undergoing cataract surgery. The amount of apoptosis detected was correlated to age, gender, type of cataract, medications and disease. Moreover, the ability of human lens epithelial cells in culture to respond to the apoptosis-inducing agent staurosporin by activation of caspase-3 was investigated. Human lens capsulotomy specimens were collected immediately after surgery, frozen and later analysed with respect to caspase-3 activity, using the fluorogenic substrate Ac-DEVD-AMC. Generally, the activity of caspase-3 detected in this manner was very low and in 23% of the specimens it was non-detectable. However, there were differences in caspase activity between lens epithelial cells from different types of cataract, where samples from lenses with posterior subcapsular cataract exhibited significantly lower caspase-3 activity than lenses with a clear subcapsular zone. Age, gender or medications did not show any correlation with caspase activity but human capsulotomy specimens from diabetic patients exhibited significantly lower caspase-3 activity. Staurosporin caused a concentration-dependent increase in caspase activity in cultured human lens epithelial cells and the amount of apoptotic nuclei was also increased as viewed by staining with Hoechst 33342, showing chromatin condensation and nuclear fragmentation. Similar results were obtained when fresh human lens capsulotomy specimens were exposed to 1000 nM staurosporin for 24 hr. To conclude, the present data indicate that human lens epithelial cells have the ability to respond to apoptosis-inducing agents with caspase-3 dependent apoptosis, and that even though the general level of apoptosis in human lens epithelium in vivo is low, there are differences in caspase-3 activity levels in lenses with or without posterior subcapsular cataract. The latter finding supports previous studies indicating that this type of cataract may result from defective differentiation, in which apoptosis may play an important role.