CellShip: An Ambient Temperature Transport and Short-Term Storage Medium for Mammalian Cell Cultures.

Cell culture is a critical platform for numerous research and industrial processes. However, methods for transporting cells are largely limited to cryopreservation, which is logistically challenging, requires the use of potentially cytotoxic cryopreservatives, and can result in poor cell recovery. Development of a transport media that can be used at ambient temperatures would alleviate these issues. In this study, we describe a novel transportation medium for mammalian cells. Five commonly used cell lines, (HEK293, CHO, HepG2, K562, and Jurkat) were successfully shipped and stored for a minimum of 72 hours and up to 96 hours at ambient temperature, after which, cells were recovered into standard culture conditions. Viability (%) and cell numbers, were examined, before, following the transport/storage period and following the recovery period. In all experiments, cell numbers returned to pretransport/storage concentration within 24-48 hours recovery. Imaging data indicated that HepG2 cells were fully adherent and had established typical growth morphology following 48 hours recovery, which was not seen in cells recovered from cryopreservation. Following recovery, Jurkat cells that had been subjected to a 96 hours transport/storage period, demonstrated a 1.93-fold increase compared with the starting cell number with >95% cell viability. We conclude that CellShip® may represent a viable method for the transportation of mammalian cells for multiple downstream applications in the Life Sciences research sector.

OECD workshop consensus report: Ethical considerations with respect to human derived products, specifically human serum, in OECD test guidelines.

The ethical needs and concerns with use and sourcing of human materials, particularly serum, in OECD in vitro test guidelines were explored in a dedicated international workshop held in 2019. The health-related aspects of the donation procedure, including tissue screening, donor health, laboratory work health protection, permission from the donor for commercial use, payment of the donors and the potential for exploitation of low-income populations and data protection of the donors; supply, availability, and competition with clinical needs; traceability of the serum and auditability/GLP needs for the Test Guideline Programme, were examined. Here we provide the recommendations of the workshop with respect to the use of human serum, and potentially other human reagents, specifically with regard to test method development for OECD Test Guideline utility as part of the Mutual Acceptance of Data requirement across all OECD member countries. These include informed donor consent terminology, a checklist of human serum information requirements to be included with the Good Laboratory Practise report, and suitable sources for human serum to ensure waste supplies are used, that can no longer be used for medical purposes, ensuring no competition of supply for essential medical use.

The Effect of Substance Abuse Programs on Positive Drug Screening Tests in Trauma Patients.

BACKGROUND: According to the National Institute on Drug Abuse, West Virginia has the highest age-adjusted rate of drug overdose deaths involving opioids. To combat this crisis, comprehensive drug counseling support services were established within the Cabell Huntington Hospital (CHH) system in October 2018 in Huntington, WV, USA. The purpose of this study was to investigate whether these services significantly reduced the number of trauma patients with a positive urine drug screen (UDS) seen at CHH. METHODS: The trauma registry at CHH was used to obtain data on trauma patients with positive UDS from January 2017 to October 2019, which was divided into groups before and after October 2018. Exclusion criteria were any patients who were prescribed the drug. The percentages of the total number of positive drug screens within each group were calculated, and a t-test analysis was performed to determine the P values. RESULTS: 345 trauma patients with positive UDS were selected. Results showed that there was an overall decrease in the rate of nonprescribed use after October 2018 of benzodiazepines (18.1%-11.5%), cocaine (19.5%-15.3%), opioids (19.1%-12.3%), and oxycodone (10.2%-4.6%). However, none of these changes were statistically significant. There was an increase in the rate of nonprescribed use of amphetamine (20.0%-23.8%) and methamphetamine (14.4%-33.8%). DISCUSSION: Our hypothesis is that the support systems are relatively new, and may need more time to identify and intervene on patients before a statistically significant effect on drug abuse rates in our region can be seen.

Differences in the perceived role of the healthcare provider in delivering vascular health checks: a Q methodology study.

BACKGROUND: The UK Department of Health introduced the National Health Service (NHS) Health Check Programme in April 2009 in an attempt to improve primary and secondary prevention of cardiovascular disease in the UK population and to reduce health inequalities. Healthcare professionals' attitudes towards giving lifestyle advice will influence how they interact with patients during consultations. We therefore sought to identify the attitudes of primary care healthcare professionals towards the delivery of lifestyle advice in the context of the NHS Health Check Programme. METHODS: Fifty-two primary care healthcare professionals undertook a Q sort with 36 statements that represented a range of viewpoints about the importance of lifestyle change, medication, giving lifestyle advice in the primary care setting, and the individual, social and material factors that might impact on lifestyle related behaviour change. Sorts were analysed by-person using principal components analysis and varimax rotation. RESULTS: Five statistically independent factors (accounts) reflected distinct views on the topic. Account 1 was supportive of initiatives like the NHS Health Check, and emphasised the importance of professionals working collaboratively with patients to facilitate lifestyle change. Account 2 expressed views on the potential overuse of statin medication and placed responsibility for lifestyle change with the patient. Account 3 viewed the healthcare professional role to be one of educator, emphasising the provision of information. Account 4 perceived lifestyle change to be difficult for patients and emphasised the need for healthcare professionals to be role models. Account 5 was inconsistent about the value of lifestyle change, or the role of healthcare professionals in promoting it, a finding that may be due to ambivalence about the health check or to lack of engagement with the Q sort task. We found no strong associations between any of the factors and, gender, role, age or ethnicity. CONCLUSIONS: Our findings suggest that healthcare professionals hold viewpoints that may influence how they interact with patients during health checks. When implementing programmes like the NHS Health Check, it would be useful to take healthcare professionals' views into account. Attitudes and beliefs could be explored during training sessions, for example.

Survey to investigate the role of the community stroke care coordinator.

The National Service Framework for older people recommends that patients recovering from a stroke should have access to a stroke care coordinator (SCC) to support their longer-term recovery. However, the knowledge base, skills and procedures for the role are poorly defined. We have therefore conducted a survey to examine the development and responsibilities of SCCs with a remit for longer-term support. We identified 39 SCCs, most of whom were nurses. Three role types were observed: enhanced community follow-up (n=22); hybrid (n=15); and hospital outpatient (n=2). However, there was overlap between the roles reflecting a wide variation in practice in respect of timing and frequency of patient contacts, and methods for assessing patients' needs. Workload and knowledge of the evidence-base were identified as the main challenges to delivering care. Meeting patients' needs was impeded by limited access to psychology and physical therapies. A more robust evidence-base to support longer-term stroke care, combined with routine clinical governance systems for monitoring care provision could enhance the development of the SCC role.

Methyl side-chain dynamics in proteins using selective enrichment with a single isotopomer.

13C relaxation studies on side-chain methyl groups in proteins typically involve measurements on (13)CHD(2) isotopomers, where the (13)C relaxation mechanism is particularly straightforward in the presence of a single proton. While such isotopomers can be obtained in proteins overexpressed in bacteria by use of (13)C enriched and fractionally deuterated media, invariably all possible (2)H isotopomers are obtained. This results in a loss of both resolution and sensitivity, which becomes particularly severe for larger proteins. We describe an approach that overcomes this problem by chemical synthesis of amino acids containing a pure (13)CHD(2) isotopomer. We illustrate the benefits of this approach in (13)C side-chain relaxation measurements on the mouse major urinary protein selectively enriched with [gamma(1),gamma(2)-(13)C(2),alpha,beta,gamma(1),gamma(1),gamma(2),gamma(2)-(2)H(6)] valine. Relaxation measurements in the absence and presence of pyrazine-derived ligands suggest that valine side-chain dynamics do not contribute significantly to binding entropy.

1H-filtered correlation experiments for assignment and determination of coupling constants in backbone labelled proteins.

The implementation of [13Calpha,13C',15N,2Halpha] labelled amino acids into proteins allows the acquisition of high resolution triple resonance experiments. We present for the first time resonance assignments facilitated by this new labelling strategy. The absence of 1JCalpha,Cbeta couplings enables us to measure 1JCalpha,C' scalar and 1DCalpha,C' residual dipolar coupling constants using modified HNCA experiments which do not suffer from sensitivity losses characteristic for 13C constant time experiments.