3DT Holdings (United States)

DNA polymerases are complex molecular machines capable of replicating genetic material using a template-driven process. While the copying function of these enzymes is well established, their ability to perform untemplated DNA synthesis is less well characterized. Here, we explore the ability of DNA polymerases to synthesize DNA fragments in the absence of a template. We use long-read nanopore sequencing, real-time fluorescence assays, and atomic force microscopy to observe the synthesis and physical structure of pools of DNA products derived from a diverse set of natural and engineered DNA polymerases across varying temperatures and buffer compositions. We detail the features of the DNA fragments generated, enrichment of select sequence motifs, and demonstrate that the sequence composition of the synthesized DNA can be altered by modifying environmental conditions. This work provides extensive data to better discern the process of untemplated DNA polymerase activity and may support its potential repurposing as a technology for the guided synthesis of DNA sequences on the kilobase-scale and beyond.

Chronic kidney disease (CKD) is traditionally viewed as a condition marked by a progressive reduction in kidney function leading to the need for kidney dialysis or transplantation. The estimated prevalence of CKD in adults in Puerto Rico is ~20% higher than that of the overall United States (US). To address the disproportionately high rate of CKD in Puerto Rico, we created a multidisciplinary chronic disease management (CDM) program targeting CKD and diabetes mellitus (DM), the leading CKD risk factor. Over 7200 eligible enrollees in a Puerto Rico-Managed Medicare Program participated in a CDM program targeting individuals with CKD or DM as determined by administrative review. Evaluations were conducted on 4068 program participants with baseline glomerular filtration rate (eGFR) and codifying CKD stage by eGFR. A dietitian/nurse team provided dietary and lifestyle recommendations to the patient/family and a nephrologist/endocrinologist made diabetes and CKD recommendations to the primary care provider. Findings on 2095 participants with Stages 1-3 CKD with follow-up eGFR at least 6 months but less than 2 years after baseline are presented. At baseline, the mean age was 74 years (range 30-101), 59% of patients were female and mean duration of follow-up from initial evaluation to second evaluation was 407 days (±159 days SD). Most participants had Stage 2 CKD (34.8%), followed by CKD Stage 1 and 3 (33.5 and 31.7%). During the follow-up period, 55.9% of participants with Stage 1 CKD remained in Stage 1, 84.9% of patients with Stage 2 remained in Stage 2 or regressed to Stage 1, while 96.1% of patients with Stage 3 remained in Stage 3 or regressed to Stage 2. Only 15.1% of patients in Stage 2 progressed to Stage 3 and 3.9% of patients in Stage 3 progressed to Stage 4 or 5. A secondary analysis comparing all 665 CDM Stage 3 participants to 117,249 historical controls found CDM participants demonstrated a higher rate of regression (20.3% vs. 15.2%; absolute difference +5.1 percentage points; p = <0.01) and a lower rate of progression (3.9% vs. 15.3%; absolute difference -11.4 percentage points; p < 0.001). Early findings of a multidisciplinary CDM intervention indicate that 79% of participants with CKD Stages 1-3 by eGFR had stabilized or improved CKD status. Comparison to a randomized control group to better assess for causality and longer-term CDM program follow-up on CKD status and clinical outcomes is warranted.

Fecobionics is a novel integrated technology for assessment of anorectal function. It is a defecatory test with simultaneous measurements of pressures, orientation, and device angle (a proxy of the anorectal angle). Furthermore, the latest Fecobionics prototypes measure diameters (shape) using impedance planimetry during evacuation of the device. The simultaneous measurement of multiple variables in the integrated test allows new metrics to be developed including more advanced novel defecation indices, enabling mechanistic insight in the defecation process at an unprecedented level in patients with anorectal disorders including patients suffering from obstructed defecation, fecal incontinence, and low anterior resection syndrome. The device has the consistency and shape of a normal stool (type 3–4 on the Bristol Stool Form Scale). Fecobionics has been validated on the bench and in animal studies and used in clinical trials to study defecation phenotypes in normal human subjects and patients with obstructed defecation, fecal incontinence, and low anterior resection syndrome after rectal cancer surgery. Subtypes have been defined, especially of patients with obstructed defecation. Furthermore, Fecobionics has been used to monitor biofeedback therapy in patients with fecal incontinence to predict the outcome of the therapy (responder versus non-responder). Most Fecobionics studies showed a closer correlation to symptoms as compared to current technologies for anorectal assessment. The present article outlines previous and ongoing work, and perspectives for future studies in proctology, including in physiological assessment of function, diagnostics, monitoring of therapy, and as a tool for biofeedback therapy.

Legalization of cannabis in the US and other countries highlight the need to understand the health consequences of this substance use. Research indicates that some cannabis ingredients may play beneficial role in treating various medical conditions while other ingredients may pose health risks. This review is focused on the brain and mental health effects of cannabis use. The rationale for examining cannabis use in behavioral and neural conditions is that these conditions are highly widespread in the US and account for high level of medical healthcare and associated cost. The purpose of this review is to provide an overview of the known medicinal benefits of selected cannabis cannabinoids in conditions like pediatric epilepsy, attention deficit hyperactivity disorder, autism spectrum disorder, and the known side effects or contraindications in conditions such as addiction, cognition, and psychosis. Several recommendations are made as to studies that will help further understanding the increasing role of cannabis in neuropsychiatric health and disease.

Abstract Prioritizing Ontario’s long-term care home (LTCH) residents for vaccination against severe acute respiratory syndrome coronavirus 2 has drastically reduced their disease burden; however, recent LTCH outbreaks of variants of concern (VOCs) have raised questions regarding their immune responses. In 198 residents, mRNA vaccine dose 1 elicited partial spike and receptor binding domain antibody responses, while the second elicited a response at least equivalent to convalescent individuals in most residents. Residents administered mRNA-1273 (Moderna) mounted stronger total and neutralizing antibody responses than those administered BNT162b2 (Pfizer-BioNTech). Two to four weeks after dose 2, residents (n= 119, median age 88) produced 4.8–6.3-fold fewer neutralizing antibodies than staff (n= 78; median age 47) against wild-type (with D614G) pseudotyped lentivirus, and residents administered BNT162b2 produced 3.89-fold fewer neutralizing antibodies than those who received mRNA-1273. These effects were exacerbated upon serum challenge with pseudotyped VOC spike, with up to 7.94-fold reductions in B.1.351 (Beta) neutralization. Cumulatively, weaker vaccine stimulation, age/comorbidities, and the VOC produced an ∼130-fold reduction in apparent neutralization titers in LTCH residents and 37.9% of BNT162b2-vaccinated residents had undetectable neutralizing antibodies to B.1.351. Continued immune response surveillance and additional vaccine doses may be required in this population with known vulnerabilities.

Data modification operations (update, delete, insert) may require significant computational power and may cause delays in the execution of other queries. We start with abstract analysis of reasons why data modification and concurrency control may affect performance of the database server and individual queries. Besed on this analysis we identify several cases that may necessitate tuning. Specifically we discuss the impact of multiple indexes, large and frequent updates, triggers and integrity constraints.

A query is considered long if a significant part of at least one large table is needed to calculate the output. The techniques needed to tune long queries differ from those for short queries. We show how to reduce the size of data being processed during the execution of a long query, how to reduce the naumber of full scans and how to control the choice of join algorithms and other decisions made by the optimizer.