rAAV2/5 Gene-Targeting to Rods: Dose-Dependent Efficiency and Complications Associated With Different Promoters

A prerequisite for using corrective gene therapy to treat humans with inherited retinal degenerative diseases that primarily affect rods is to develop viral vectors that target specifically this population of photoreceptors. The delivery of a viral vector with photoreceptor tropism coupled with a rod-specific promoter is likely to be the safest and most efficient approach to target expression of the therapeutic gene to rods. Three promoters that included a fragment of the proximal mouse opsin promoter (mOP), the human G-protein-coupled receptor protein kinase 1 promoter (hGRK1), or the cytomegalovirus immediate early enhancer combined with the chicken β actin proximal promoter CBA were evaluated for their specificity and robustness in driving GFP reporter gene expression in rods, when packaged in a recombinant adeno-associated viral vector of serotype 2/5 (AAV2/5), and delivered via subretinal injection to the normal canine retina. Photoreceptor-specific promoters (mOP, hGRK1) targeted robust GFP expression to rods, whereas the ubiquitously expressed CBA promoter led to transgene expression in the retinal pigment epithelium, rods, cones and rare Müller, horizontal and ganglion cells. Late onset inflammation was frequently observed both clinically and histologically with all three constructs when the highest viral titers were injected. Cone loss in the injected regions of the retinas that received the highest titers occurred with both the hGRK1 and CBA promoters. Efficient and specific rod transduction, together with preservation of retinal structure was achieved with both mOP and hGRK1 promoters when viral titers in the order of 1011 vg ml–1 were used

Copper transformation, speciation, and detoxification in anoxic and suboxic freshwater sediments

The complex chemistry of copper (Cu) in freshwater sediments at low concentrations is not well understood. We evaluated the transformation processes of Cu added to freshwater sediments under suboxic and anoxic conditions. Freshwater sediments from three sources in Michigan with different characteristics (Spring Creek, River Raisin, and Maple Lake) were spiked with 30 or 60 mg kg-1 Cu and incubated under a nitrogen atmosphere. After 28-d, each treatment subset was amended with organic matter (OM) to promote anoxic conditions and evaluate its effects on Cu speciation. OM addition triggered a shift from suboxic to anoxic conditions, and sequential extractions showed that Cu accordingly shifted from acid-soluble to oxidizable fractions. Extended X-ray absorption fine-structure (EXAFS) spectroscopy revealed that Cu sulfides dominated all anoxic samples except for Spring Creek 30 mg kgxfffd; 1, where Cu(I) was predominantly complexed to thiol groups of OM. Covellite and chalcopyrite (CuFeS2) were the predominant Cu species in nearly all anoxic samples, as determined by Raman spectroscopy, scanning electron microscopy, and X-ray absorption near-edge structure (XANES) spectroscopy. Copper reduction also occurred under suboxic conditions: for two of three sediments, around 80% had been reduced to Cu(I), while the remaining 20% persisted as Cu(II) complexed to OM. However, in the third coarsest (i.e., Spring Creek), around 50% of the Cu had been reduced, forming Cu(I)-OM complexes, while the remainder was Cu(II)-OM complexes. Toxicity tests showed that survival of H. azteca and D. magna were significantly lower in suboxic treatments. Anoxic sediments triggered a near-complete transformation of Cu to sulfide minerals, reducing its toxicity

Histologic and bacteriologic findings in valvular endocarditis of slaughter-age pigs

Endocarditis lesions from 117 slaughter pigs were examined pathologically and etiologically in addition to 90 control hearts with cardiac valves. Lesions were located on the valves; however, the lesions had extended to the walls in 21 cases (18%). Lesions predominated on the mitral valve (59%). A total of 28 cases, from which no growth was obtained or a contamination flora was grown, were screened by fluorescence in situ hybridization (FISH) for bacteria (general bacterial probe) and probes specific for Streptococcus suis and Erysipelothrix rhusiopathiae, respectively. Using FISH, an additional 10 cases of endocarditis due to S. suis and E. rhusiopathiae were disclosed. Within lesions, streptococci predominated (53%) followed by E. rhusiopathiae (30%). Distinct features of both the lesions and the shape and localization of bacterial colonies were related to streptococci and E. rhusiopathiae. The propensity for streptococci to be localized on more than 1 valve in single hearts may be because S. suis–infected pigs tend to have been infected for a longer period compared with E. rhusiopathiae. Mineralization of endocarditis lesions was significantly associated with infection by streptococci, and was seen in 71% of the cases, whereas it was present in only 28% of lesions caused by E. rhusiopathiae. In addition, areas with mineralization were significantly correlated to the presence of a granulomatous reaction. Granulomatous endocarditis is likely a result of a foreign body reaction due to dystrophic mineralization. Local proliferation of valvular endothelial cells, found in 9 hearts in the current study, may increase the risk of developing thrombosing endocarditis in pigs. </jats:p

SARM1 depletion rescues NMNAT1-dependent photoreceptor cell death and retinal degeneration

Leber congenital amaurosis type nine is an autosomal recessive retinopathy caused by mutations of the NA

The Real-World Observational Prospective Study of Health Outcomes with Dulaglutide and Liraglutide in Type 2 Diabetes Patients (TROPHIES): Baseline Patient-Reported Outcomes

Introduction: Although patient-reported outcome (PRO) measures provide important information beyond clinical data, studies that assess the PROs of type 2 diabetes mellitus (T2DM) patients initiating injectable glucose-lowering medications in routine clinical practice are limited. We describe the perspectives of patients based on a diversified panel of generic and disease-specific PRO measures at the time of enrollment (baseline) in the TROPHIES study. Methods: TROPHIES is a 24-month prospective observational study performed in France, Germany, and Italy in patients with T2DM who initiated their first injectable glucose-lowering medication with once-weekly dulaglutide or once-daily liraglutide. To better understand the perspectives of these patients regarding their overall health, treatment satisfaction, and quality of life and work, the patients’ responses to the following questionnaires were collected at baseline before they initiated treatment with dulaglutide or liraglutide: EQ-5D-5L (scale: 0–1), EQ-VAS (visual analog scale: 0–100), Impact of Weight on Self-Perceptions Questionnaire (IW-SP; scale: 0–100), Diabetes Treatment Satisfaction Questionnaire Status (DTSQs; scale: 0–36), and Diabetes Productivity Measure (DPM; scale: 0–100). Analyses were descriptive in nature, with higher scores reflecting better outcomes. Results: Data from patients at the time of enrollment were analyzed. At baseline, patients initiating dulaglutide (N = 1130) or liraglutide (N = 1051) rated their quality of life in terms of mean EQ-5D-5L index as 0.84 and 0.83, and in terms of mean EQ-VAS as 67.5 and 67.5, respectively. The mean baseline scores in patients initiating dulaglutide or liraglutide were 59.8 and 61.3 for IW-SP, 24.6 and 25.8 for DTSQs, 78.6 and 79.5 for DPM Life Productivity, and 87.5 and 86.8 for DPM Work Productivity, respectively. Conclusion: The information from this varied panel of PRO instruments collected at baseline complements clinical outcomes data

Using an online collaboration platform to facilitate group work

The onset of COVID-19 necessitated moving three large-enrolment introductory engineering and IT subjects online after just one week of face to face teaching. All three subjects focus on facilitating students' learning through group work to solve a self-identified problem. Considering a key Subject Learning Outcome is 'to collaborate effectively in team processes', group work is integral to the aims of these subjects. Studies for both online and face-to-face group work identify the influence educators play in achieving successful learning outcomes and group satisfaction; for example, the importance of group work management (Xu, Du & Fan, 2015). While many challenges faced in online group work in education are common to face-to-face teaching (Roberts & McInnerney, 2007), it has been shown that "distance does matter" (Olson & Olson, 2000). The challenge was to facilitate the same level of cooperation between students and enable them to build teamwork skills without face-to-face interaction with teammates or educators. PURPOSE OR GOAL Moving three early-year subjects of approximately 600 students each onto an online collaboration platform over a short period provided new challenges. This paper will discuss the aspects of our transition to online group work that worked well, and those that did not, from the perspective of students and tutors. These insights into best-practice online learning will inform how teaching can shift into blended learning in 2021. METHODOLOGY Focus groups were conducted with students from one second-year and two first-year subjects. In these focus groups, students discussed their experiences of working in a group environment and how this experience can be improved. The comments from student feedback surveys and students’ comments from the SPARKplus peer assessments were also used. In addition, feedback on classes and materials were taken from tutors and compared to the students' view of the online classes. Transcripts and comments were analysed to identify recurrent themes. OUTCOMES Students had mixed feelings about the transition to online classes. Despite the use of many different strategies, issues with engagement remain. However, some strategies have emerged that motivate the students, while successful ways for groups to work together were discovered. Our research also indicates that the use of a learning platform which facilitates instant student discussion is worth further investigation for online classes. CONCLUSIONS/RECOMMENDATIONS/SUMMARY Online classes make it more challenging to engage students in group work based learning. However, careful fostering of group team spirit helps to lessen individual isolation. Some students thrive from the provision of instant feedback through an interactive learning platform. In future work, focus groups need to be conducted with students who achieved lower marks or dropped out of the subject to understand the issues they faced. More attention needs to be paid to the tutor perspective and how activities can be better adapted to student needs. KEYWORDS Transition online, group work, student engagement

The Real-World Observational Prospective Study of Health Outcomes with Dulaglutide and Liraglutide in Type 2 Diabetes Patients (TROPHIES): Design and Baseline Characteristics

Introduction: The TROPHIES observational study enrolled patients with type&nbsp;2 diabetes mellitus (T2DM) initiating their first injectable treatment with the glucagon-like peptide&nbsp;1 receptor agonists (GLP-1&nbsp;RAs) dulaglutide or liraglutide. This manuscript focuses on the study design, baseline characteristics of the enrolled population, and factors associated with GLP-1&nbsp;RA choice. Methods: TROPHIES is a prospective, observational, 24-month study conducted in France, Germany, and Italy. Inclusion criteria include adult patients with T2DM, naïve to injectable antihyperglycemic treatments, initiating dulaglutide or liraglutide per routine clinical practice. The primary outcome is the duration of treatment on dulaglutide or liraglutide without a significant treatment change. Results: The analysis included 2181 patients (dulaglutide, 1130; liraglutide, 1051) (cutoff date May&nbsp;15, 2019). The population was 56% male with mean [standard deviation (SD)] patient characteristics at baseline as follows: age, 59.2 (11.0) years; body mass index (BMI), 33.9 (6.6) kg/m2; T2DM duration, 8.5 (6.9) years; and glycated hemoglobin (HbA1c), 8.2 (1.3)%. Between-cohort demographic and clinical characteristics were balanced. The mean (SD) HbA1c and BMI values for French, German, and Italian patients were, respectively, 8.6 (1.4)%, 8.2 (1.4)%, 8.0 (0.8)%; 33.3 (6.1) kg/m2, 36.0 (7.2) kg/m2, and 32.6 (5.9) kg/m2. Conclusion: This study analysis at baseline provides an opportunity to evaluate between-country differences in baseline HbA1c, weight, macrovascular complications, and factors driving GLP-1&nbsp;RA selection for patients with T2DM in daily practice

A Telehealth-Delivered Pulmonary Rehabilitation Intervention in Underserved Hispanic and African American Patients With Chronic Obstructive Pulmonary Disease: A Community-Based Participatory Research Approach.

BACKGROUND:Although home telemonitoring (TM) is a promising approach for patients managing their chronic disease, rehabilitation using home TM has not been tested for use with individuals living with chronic obstructive pulmonary disease (COPD) residing in underserved communities. OBJECTIVE:This study aimed to analyze qualitative data from focus groups with key stakeholders to ensure the acceptability and usability of the TM COPD intervention. METHODS:We utilized a community-based participatory research (CBPR) approach to adapt a home TM COPD intervention to facilitate acceptability and feasibility in low-income African American and Hispanic patients. The study engaged community stakeholders in the process of modifying the intervention in the context of 2 community advisory board meetings. Discussions were audio recorded and professionally transcribed and lasted approximately 2 hours each. Structural coding was used to mark responses to topical questions in interview guides. RESULTS:We describe herein the formative process of a CBPR study aimed at optimizing telehealth utilization among African American and Latino patients with COPD from underserved communities. A total of 5 major themes emerged from qualitative analyses of community discussions: equipment changes, recruitment process, study logistics, self-efficacy, and access. The identification of themes was instrumental in understanding the concerns of patients and other stakeholders in adapting the pulmonary rehabilitation (PR) home intervention for acceptability for patients with COPD from underserved communities. CONCLUSIONS:These findings identify important adaptation recommendations from the stakeholder perspective that should be considered when implementing in-home PR via TM for underserved COPD patients. TRIAL REGISTRATION:ClinicalTrials.gov NCT03007485; https://clinicaltrials.gov/ct2/show/NCT03007485

High-throughput screening of tick-borne pathogens in Europe

Due to increased travel, climatic, and environmental changes, the incidence of tick-borne disease in both humans and animals is increasing throughout Europe. Therefore, extended surveillance tools are desirable. To accurately screen tick-borne pathogens (TBPs), a large scale epidemiological study was conducted on 7050 Ixodes ricinus nymphs collected from France, Denmark, and the Netherlands using a powerful new high-throughput approach. This advanced methodology permitted the simultaneous detection of 25 bacterial, and 12 parasitic species (including; Borrelia, Anaplasma, Ehrlichia, Rickettsia, Bartonella, Candidatus Neoehrlichia, Coxiella, Francisella, Babesia, and Theileria genus) across 94 samples. We successfully determined the prevalence of expected (Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Rickettsia helvetica, Candidatus Neoehrlichia mikurensis, Babesia divergens, Babesia venatorum), unexpected (Borrelia miyamotoi), and rare (Bartonella henselae) pathogens in the three European countries. Moreover we detected Borrelia spielmanii, Borrelia miyamotoi, Babesia divergens, and Babesia venatorum for the first time in Danish ticks. This surveillance method represents a major improvement in epidemiological studies, able to facilitate comprehensive testing of TBPs, and which can also be customized to monitor emerging diseases