The Application of Clinical Lithotripter Shock Waves to RNA Nucleotide Delivery to Cells

AbstractThe delivery of genes into cells through the transfer of ribonucleic acids (RNAs) has been found to cause a change in the level of target protein expression. RNA-based transfection is conceptually more efficient than commonly delivered plasmid DNA because it does not require division or damage of the nuclear envelope, thereby increasing the chances of the cell remaining viable. Shock waves (SWs) have been found to induce cellular uptake by transiently altering the permeability of the plasma membrane, thereby overcoming a critical step in gene therapy. However, accompanying SW bio-effects include dose-dependent irreversible cell injury and cytotoxicity. Here, the effect of SWs generated by a clinical lithotripter on the viability and permeabilisation of three different cell lines in vitro was investigated. Comparison of RNA stability before and after SW exposure revealed no statistically significant difference. Optimal SW exposure parameters were identified to minimise cell death and maximise permeabilisation, and applied to enhanced green fluorescent protein (eGFP) messenger RNA (mRNA) or anti-eGFP small interfering RNA delivery. As a result, eGFP mRNA expression levels increased up to 52-fold in CT26 cells, whereas a 2-fold decrease in GFP expression was achieved after anti-eGFP small interfering RNA delivery to MCF-7/GFP cells. These results indicate that SW parameters can be employed to achieve effective nucleotide delivery, laying the foundation for non-invasive and high-tolerability RNA-based gene therapy

Effects of telemonitoring on glycaemic control and healthcare costs in type 2 diabetes: a randomised controlled trial

Introduction: This study examined the effect of a telehealth intervention on the control of type 2 diabetes, and subsequent potential cost-savings to the health system. Methods: This prospective randomised controlled trial randomised adults with type 2 diabetes to the intervention (diabetes program) or control (usual care) arm. Key eligibility criteria included an HbA1c level of at least 58 mmol/mol (7.5%) without severe or unstable comorbidities. All participants continued their usual healthcare, but participants in the intervention arm received additional diabetes care from a diabetes care coordinator via a home monitor that captured clinical measures. Data collected included biomedical, quality of life measures and healthcare (GP, outpatient and inpatient) costs. The primary outcome was HbA1c collected at baseline and six-months. Analysis was conducted on a complete case intention-to-treat basis. The healthcare system perspective was taken to calculate the incremental cost per percentage-point reduction in HbA1c. Results: Results from 63 participants from each study arm were analysed. HbA1c in the intervention group decreased from a median 68 mmol/mol (8.4%) to 58 mmol/mol (7.5%), and remained unchanged in the control group at median 65 mmol/mol (8.1%) at the six-month endpoint. The intervention effect on HbA1c change was statistically significant (p=0.004). Total healthcare costs in the intervention group, including the intervention costs, were lower (mean $3,781 vs$4,662; p<0.001) compared to usual care. Discussion: There was a clinically meaningful and statistically significant benefit from the telehealth intervention at a lower cost; thus, telehealth was cost-saving and produced greater health benefits compared to usual care

A systems approach to the perception of the integration of public health into pharmacy practice: A qualitative study

Background: Pharmacists, as health professionals, are ideally positioned to support the health and wellbeing of populations, in addition to their role of providing individualised health care. Objective: The aim of this study was to explore current opinion on the contribution of pharmacists to public health and how this may be enhanced to positively impact public health indicators. Methods: A total of 24 pharmacists from Australia, United Kingdom, Canada and the United States of America, and Australian public health professionals and consumers participated in semi-structured interviews between January and October 2021. Interpretive thematic analysis was applied with coding of transcripts using the constant comparison method. Themes were developed and named in accordance with Bronfenbrenner’s ecological theory of development. Results: Pharmacists contribute to public health and have important roles in health education and illness prevention services. Strong enablers in community pharmacy include trust by consumers and ease of accessibility to pharmacists. Pharmacists are viewed as leaders in communities and contribute to the health system broadly in areas such as medication policy and public health organisations. Participants suggested that pharmacist contributions to public health are often unrecognized by the pharmacy profession, health professionals and consumers, and could be developed to allow effective contributions. Strategies to improve pharmacist contributions included clarifying public health-related terminology, increased development of pharmacy roles and reform for community pharmacies to participate in health prevention and promotion services. Integration of public health in pharmacy education, professional development, and recognition of pharmacy roles across all system levels were also identified as important. Conclusions: The study indicated that pharmacists currently contribute to the improvement of public health. However, development strategies are required for this to be more effective in integrating public health approaches into their professional practice to be recognized for their public health-related roles

The UK quality and outcomes framework pay-for-performance scheme and spirometry: rewarding quality or just quantity? A cross-sectional study in Rotherham, UK

Background: Accurate spirometry is important in the management of COPD. The UK Quality and Outcomes Framework pay-for-performance scheme for general practitioners includes spirometry related indicators within its COPD domain. It is not known whether high achievement against QOF spirometry indicators is associated with spirometry to BTS standards. Methods: Data were obtained from the records of 3,217 patients randomly sampled from 5,649 patients with COPD in 38 general practices in Rotherham, UK. Severity of airflow obstruction was categorised by FEV1 (% predicted) according to NICE guidelines. This was compared with clinician recorded COPD severity. The proportion of patients whose spirometry met BTS standards was calculated in each practice using a random sub-sample of 761 patients. The Spearman rank correlation between practice level QOF spirometry achievement and performance against BTS spirometry standards was calculated. Results: Spirometry as assessed by clinical records was to BTS standards in 31% of cases (range at practice level 0% to 74%). The categorisation of airflow obstruction according to the most recent spirometry results did not agree well with the clinical categorisation of COPD recorded in the notes (Cohen's kappa = 0.34, 0.30 - 0.38). 12% of patients on COPD registers had FEV1 (% predicted) results recorded that did not support the diagnosis of COPD. There was no association between quality, as measured by adherence to BTS spirometry standards, and either QOF COPD9 achievement (Spearman's rho = -0.11), or QOF COPD10 achievement (rho = 0.01). Conclusion: The UK Quality and Outcomes Framework currently assesses the quantity, but not the quality of spirometry

Signal Transmission in the Auditory System

Contains table of contents for Section 3, an introduction and reports on six research projects.Health Sciences FundNational Institutes of Health Grant 5 R01 DC00194National Institutes of Health Grant 8 P01 DC00119National Institutes of Health Grant 5 R01 DC00473National Institutes of Health Grant 5 R01 DC00238National Institutes of Health Grant 5 T32 DC00006National Institutes of Health Grant 5 P01 DC00361National Institutes of Health Grant 5 R01 DC00235Peoples Republic of China FellowshipUnisys Corporation Doctoral FellowshipWhitaker Health Sciences Fellowshi

Discovery of widespread transcription initiation at microsatellites predictable by sequence-based deep neural network

Using the Cap Analysis of Gene Expression (CAGE) technology, the FANTOM5 consortium provided one of the most comprehensive maps of transcription start sites (TSSs) in several species. Strikingly, ~72% of them could not be assigned to a specific gene and initiate at unconventional regions, outside promoters or enhancers. Here, we probe these unassigned TSSs and show that, in all species studied, a significant fraction of CAGE peaks initiate at microsatellites, also called short tandem repeats (STRs). To confirm this transcription, we develop Cap Trap RNA-seq, a technology which combines cap trapping and long read MinION sequencing. We train sequence-based deep learning models able to predict CAGE signal at STRs with high accuracy. These models unveil the importance of STR surrounding sequences not only to distinguish STR classes, but also to predict the level of transcription initiation. Importantly, genetic variants linked to human diseases are preferentially found at STRs with high transcription initiation level, supporting the biological and clinical relevance of transcription initiation at STRs. Together, our results extend the repertoire of non-coding transcription associated with DNA tandem repeats and complexify STR polymorphism

A qualitative case study of telehealth for in-home monitoring to support the management of type 2 diabetes

The present study formed part of a randomised controlled trial of telehealth for in-home monitoring to support people with poorly controlled type 2 diabetes. We explored the experiences of patients and healthcare practitioners, and their perceptions of the telehealth model of care used in the trial. In addition to their usual diabetes care, participants receive diabetes care from a diabetes educator nurse via an in-home broadband communication device. On average, each patient participated in 14 videoconferences with a diabetes care coordinator during the 12-month trial period. Qualitative data was collected from two general practices and included semi-structured interviews and document review of patient clinical notes. A total of 12 people were interviewed: 8 health practitioners and 4 patients. Patients and health practitioners expressed a high level of satisfaction with the model of care provided. Patients also reported positive health and social outcomes as a result of being involved in the trial and indicated that in the main they had achieved their goals and were happy with their progress over the 12-month period. Analysis of interviews revealed three broad elements associated with the implementation of telehealth: interpersonal factors, operational problems and the wider health system context within which the general practices and trial team were operating. The findings suggest that adopting telehealth in the management of type 2 diabetes can lead to improved diabetes control, but more support is required to ensure sustainability and widespread implementation

The Application of Clinical Lithotripter Shock Waves to RNA Nucleotide Delivery to Cells.

The delivery of genes into cells through the transfer of ribonucleic acids (RNAs) has been found to cause a change in the level of target protein expression. RNA-based transfection is conceptually more efficient than commonly delivered plasmid DNA because it does not require division or damage of the nuclear envelope, thereby increasing the chances of the cell remaining viable. Shock waves (SWs) have been found to induce cellular uptake by transiently altering the permeability of the plasma membrane, thereby overcoming a critical step in gene therapy. However, accompanying SW bio-effects include dose-dependent irreversible cell injury and cytotoxicity. Here, the effect of SWs generated by a clinical lithotripter on the viability and permeabilisation of three different cell lines in vitro was investigated. Comparison of RNA stability before and after SW exposure revealed no statistically significant difference. Optimal SW exposure parameters were identified to minimise cell death and maximise permeabilisation, and applied to enhanced green fluorescent protein (eGFP) messenger RNA (mRNA) or anti-eGFP small interfering RNA delivery. As a result, eGFP mRNA expression levels increased up to 52-fold in CT26 cells, whereas a 2-fold decrease in GFP expression was achieved after anti-eGFP small interfering RNA delivery to MCF-7/GFP cells. These results indicate that SW parameters can be employed to achieve effective nucleotide delivery, laying the foundation for non-invasive and high-tolerability RNA-based gene therapy

Miscarriages following caesarean section-induced extra-uterine adhesions

A Caucasian lady in her 30s presented with repeated miscarriages following a previous caesarean section. Following ongoing pain and bleeding, and inconclusive imaging, a scan undertaken after her second miscarriage suggested a possible communication through the uterine wall. Subsequent laparoscopy revealed a dense band of adhesions starting from lower segment of uterus and extending all the way to anterior fundus adherent to the anterior abdominal wall. To our knowledge, this is the first published case of repeated miscarriages following extra-uterine adhesions post caesarean section. Although causality cannot be proven, this is an important and potentially treatable diagnosis to consider