Effects of preservation method on canine (Canis lupus familiaris) fecal microbiota.

Studies involving gut microbiome analysis play an increasing role in the evaluation of health and disease in humans and animals alike. Fecal sampling methods for DNA preservation in laboratory, clinical, and field settings can greatly influence inferences of microbial composition and diversity, but are often inconsistent and under-investigated between studies. Many laboratories have utilized either temperature control or preservation buffers for optimization of DNA preservation, but few studies have evaluated the effects of combining both methods to preserve fecal microbiota. To determine the optimal method for fecal DNA preservation, we collected fecal samples from one canine donor and stored aliquots in RNAlater, 70% ethanol, 50:50 glycerol:PBS, or without buffer at 25 °C, 4 °C, and -80 °C. Fecal DNA was extracted, quantified, and 16S rRNA gene analysis performed on Days 0, 7, 14, and 56 to evaluate changes in DNA concentration, purity, and bacterial diversity and composition over time. We detected overall effects on bacterial community of storage buffer (F-value = 6.87, DF = 3, P < 0.001), storage temperature (F-value=1.77, DF = 3, P = 0.037), and duration of sample storage (F-value = 3.68, DF = 3, P < 0.001). Changes in bacterial composition were observed in samples stored in -80 °C without buffer, a commonly used method for fecal DNA storage, suggesting that simply freezing samples may be suboptimal for bacterial analysis. Fecal preservation with 70% ethanol and RNAlater closely resembled that of fresh samples, though RNAlater yielded significantly lower DNA concentrations (DF = 8.57, P < 0.001). Although bacterial composition varied with temperature and buffer storage, 70% ethanol was the best method for preserving bacterial DNA in canine feces, yielding the highest DNA concentration and minimal changes in bacterial diversity and composition. The differences observed between samples highlight the need to consider optimized post-collection methods in microbiome research

Distance Education And Learning Styles: Some Interesting Results

The rush to offer web courses has left many questions as to what makes them effective and satisfactory. Important issues are: the perceived advantages of web-based courses; appropriate pedagogical tools (e.g., streaming media, PowerPoint presentations) to facilitate student-content interaction, media richness, and the influence of antecedent characteristics, such as gender, experience, motivation, and learning styles. Research here indicates support for media richness, web-based course advantages, some pedagogies, and minimal support for learning styles, at least Gardner’s multiple intelligences and Keirsey’s personality styles

A duchenne muscular dystrophy gene hot spot mutation in dystrophin-deficient Cavalier King Charles Spaniels is amenable to exon 51 skipping

BACKGROUND Duchenne muscular dystrophy (DMD), which afflicts 1 in 3500 boys, is one of the most common genetic disorders of children. This fatal degenerative condition is caused by an absence or deficiency of dystrophin in striated muscle. Most affected patients have inherited or spontaneous deletions in the dystrophin gene that disrupt the reading frame resulting in unstable truncated products. For these patients, restoration of the reading frame via antisense oligonucleotide-mediated exon skipping is a promising therapeutic approach. The major DMD deletion "hot spot" is found between exons 45 and 53, and skipping exon 51 in particular is predicted to ameliorate the dystrophic phenotype in the greatest number of patients. Currently the mdx mouse is the most widely used animal model of DMD, although its mild phenotype limits its suitability in clinical trials. The Golden Retriever muscular dystrophy (GRMD) model has a severe phenotype, but due to its large size, is expensive to use. Both these models have mutations in regions of the dystrophin gene distant from the commonly mutated DMD "hot spot". METHODOLOGY/PRINCIPAL FINDINGS Here we describe the severe phenotype, histopathological findings, and molecular analysis of Cavalier King Charles Spaniels with dystrophin-deficient muscular dystrophy (CKCS-MD). The dogs harbour a missense mutation in the 5' donor splice site of exon 50 that results in deletion of exon 50 in mRNA transcripts and a predicted premature truncation of the translated protein. Antisense oligonucleotide-mediated skipping of exon 51 in cultured myoblasts from an affected dog restored the reading frame and protein expression. CONCLUSIONS/SIGNIFICANCE Given the small size of the breed, the amiable temperament and the nature of the mutation, we propose that CKCS-MD is a valuable new model for clinical trials of antisense oligonucleotide-induced exon skipping and other therapeutic approaches for DMD

Effect of Acute Posttransplant Renal Failure on the Survival of Perfused Cadaver Kidneys

Between 7973 and 7977 we encountered 22 cases of acute renal failure after transplantation in 70 patients who received perfused cadaver kidneys. Nearly two-thirds of 76 nonfunctioning grafts were lost due to subsequent superimposed rejection, often undetected and, hence, untreated. Thirty-one percent of the 76 recovered function. The recovery rate, we believe, can be improved by earlier diagnosis and treatment of rejection and by avoiding invasive diagnostic procedures in the early postoperative period. If the oliguric period extends beyond two weeks, a closed percutaneous renal biopsy is justified. The diagnosis of rejection and/or other abnormality as well as subsequent treatment are very important in these patients

Commentary on key aspects of fecal microbiota transplantation in small animal practice

The gastrointestinal tract of dogs, cats, and other mammals including humans harbors millions of beneficial microorganisms that regulate and maintain health. Fecal microbiota transplantation (FMT) is a procedure involving the administration of a fecal infusion from a healthy individual (donor) to a patient with disease to help improve health. Despite the effectiveness of FMT to treat intestinal disorders in humans, in particular recurrent Clostridium difficile infection, there is a paucity of scientific data regarding the application of FMT in veterinary patients. Here, we outline key aspects of FMT in small animal practice

Implementation of Multipronged Approach in Patients with Chest Trauma Reduces VAP and Unplanned Admission to the ICU

Problem The incidence of pulmonary complications in trauma patients with chest trauma has been reported to be as high as 49% (Ruibel L, 2022). However, all trauma patients are at risk for developing pulmonary complications due to a multitude of factors some pre-existing while others injury related sequela. Within our institution, we observed an increase in pulmonary complications through review of our TQIP (Spring 2020) and state registry data.https://jdc.jefferson.edu/surgeryposters/1013/thumbnail.jp

Emotion, deliberation, and the skill model of virtuous agency

A recent skeptical challenge denies deliberation is essential to virtuous agency: what looks like genuine deliberation is just a post hoc rationalization of a decision already made by automatic mechanisms (Haidt 2001; Doris 2015). Annas’s account of virtue seems well-equipped to respond: by modeling virtue on skills, she can agree that virtuous actions are deliberation-free while insisting that their development requires significant thought. But Annas’s proposal is flawed: it over-intellectualizes deliberation’s developmental role and under-intellectualizes its significance once virtue is acquired. Doing better requires paying attention to a distinctive form of anxiety—one that functions to engage deliberation in the face of decisions that automatic mechanisms alone cannot resolve

Fuzapladib in a randomized controlled multicenter masked study in dogs with presumptive acute onset pancreatitis

We read with interest the article by Steiner et al,1 that claims that administration of fuzapladib is safe and effective in reducing 2 clinical scores in dogs with acute pancreatitis (AP). We commend Steiner et al for their efforts in addressing a critical need in veterinary medicine. This letter, however, raises significant concerns regarding the methodology and interpretation of the study results

Pre-operative self-efficacy education vs. usual care for patients undergoing joint replacement surgery: a pilot randomised controlled trial

Background Hip and knee replacement is a major surgical procedure performed worldwide. Despite 20 or so years of clinical research and care guidelines, the management of acute postoperative pain continues to be a concern. A growing number of self-efficacy strategies are being included in education programs for patients to enable then to have a central role in managing their illness and symptoms. Aims and Objectives The purpose of this pilot study was to evaluate the feasibility of testing an education intervention to improve self-efficacy in patients undergoing hip or knee replacement. Methods A single-blinded, parallel, pilot randomised control trial design was used. Ninety-one patients undergoing hip or knee replacement surgery were randomly assigned to an intervention or control group. Intervention group participants were given a DVD demonstrating self-efficacy activities to undertake four times before admission. Feasibility criteria related to recruitment, protocol adherence and missing data were assessed. Participants were assessed for pain, anxiety, self-efficacy and healthcare utilisation. Results In relation to recruitment, 55% of screened patients were eligible and of these 81% enrolled (n = 91). Exclusion following randomisation was 10% with missing data ranging from 0 to 20.7%. Nineteen per cent of participants were lost to follow up in the control group and 20% lost to follow up in DVD group. Protocol adherence to components of the intervention varied. Both groups were generally satisfied with pain management during hospitalisation, and there were no differences in groups on clinical outcome measures. Conclusions Preliminary evidence for the benefits of self-efficacy-based education for patients undergoing hip or knee replacement was identified. Additional findings included a need to strengthen the intervention and reducing the number of data collection points to improve the protocol, missing data and numbers lost to follow up before a larger trial is undertaken

How do people live life successfully with Parkinson's disease?

The aim of this paper is to explore how people live life successfully with Parkinson's disease and what contributed to the level of success