Isolation and Characterization of Neutralizing Monoclonal Antibodies from a Large Panel of Murine Antibodies against RBD of the SARS-CoV-2 Spike Protein.

The COVID-19 pandemic, once a global crisis, is now largely under control, a testament to the extraordinary global efforts involving vaccination and public health measures. However, the relentless evolution of SARS-CoV-2, leading to the emergence of new variants, continues to underscore the importance of remaining vigilant and adaptable. Monoclonal antibodies (mAbs) have stood out as a powerful and immediate therapeutic response to COVID-19. Despite the success of mAbs, the evolution of SARS-CoV-2 continues to pose challenges and the available antibodies are no longer effective. New variants require the ongoing development of effective antibodies. In the present study, we describe the generation and characterization of neutralizing mAbs against the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein by combining plasmid DNA and recombinant protein vaccination. By integrating genetic immunization for rapid antibody production and the potent immune stimulation enabled by protein vaccination, we produced a rich pool of antibodies, each with unique binding and neutralizing specificities, tested with the ELISA, BLI and FACS assays and the pseudovirus assay, respectively. Here, we present a panel of mAbs effective against the SARS-CoV-2 variants up to Omicron BA.1 and BA.5, with the flexibility to target emerging variants. This approach ensures the preparedness principle is in place to address SARS-CoV-2 actual and future infections.

An EAPC white paper on multi-disciplinary education for spiritual care in palliative care.

BACKGROUND: The EAPC White Paper addresses the issue of spiritual care education for all palliative care professionals. It is to guide health care professionals involved in teaching or training of palliative care and spiritual care; stakeholders, leaders and decision makers responsible for training and education; as well as national and local curricula development groups. METHODS: Early in 2018, preliminary draft paper was written by members of the European Association for Palliative Care (EAPC) spiritual care reference group inviting comment on the four core elements of spiritual care education as outlined by Gamondi et al. (2013) in their paper on palliative care core competencies. The preliminary draft paper was circulated to experts from the EAPC spiritual care reference group for feedback. At the second stage feedback was incorporated into a second draft paper and experts and representatives of national palliative care organizations were invited to provide feedback and suggest revisions. The final version incorporated the subsequent criticism and as a result, the Gamondi framework was explored and critically revised leading to updated suggestions for spiritual care education in palliative care. RESULTS: The EAPC white paper points out the importance of spiritual care as an integral part of palliative care and suggests incorporating it accordingly into educational activities and training models in palliative care. The revised spiritual care education competencies for all palliative care providers are accompanied by the best practice models and research evidence, at the same time being sensitive towards different development stages of the palliative care services across the European region. CONCLUSIONS: Better education can help the healthcare practitioner to avoid being distracted by their own fears, prejudices, and restraints and attend to the patient and his/her family. This EAPC white paper encourages and facilitates high quality, multi-disciplinary, academically and financially accessible spiritual care education to all palliative care staff.

Improving the ecological relevance of aquatic bacterial communities in biodegradability screening assessments.

Concentrating cells from aqueous samples is a common requirement for the enumeration of biomass, investigations of microbial diversity and detection of relatively rare organisms in the environment. Accurately representing the initial sampled environments in the concentrated cells is of particular importance when the subsequent analyses have tangible environmental, economic and societal consequences, as is the case with environmental exposure and risk assessment of chemicals. This study investigated the potential use of four different cell concentration methods: centrifugation, membrane filtration, tangential flow filtration and column colonisation. These methods were assessed against a series of scientific and practical criteria, including: similarity of concentrated community to initial environmental sample; cell concentration achieved; biodegradation test outcome; sample throughput; and capital and maintenance costs. All methods increased cell concentration by as little as 10-fold to as much as 1000-fold. DGGE and 454 pyrosequencing analysis showed concentrated communities to have >60% similarity to each other, and the initial sample. There was a general trend for a more reliable assessment of 4-nitrophenol biodegradation in 96-well plate biodegradation assays, with increasing cell concentration. Based on the selection criteria, it is recommended that there is not one concentration method fit for all purposes, rather, the appropriate method should be selected on a case-by-case basis. Membrane filtration would be the most suitable method for low sample volumes; the increased throughput capacity of tangential flow filtration renders it most suitable for large volumes; and centrifugation is most suitable for samples with high initial biomass concentrations. The poor similarity in microbial community composition of the column colonised samples compared to the initial samples, suggested a concentration basis; this combined with its low sample throughput precluded this approach for future concentration studies of planktonic bacterial samples.

High Throughput Biodegradation-Screening Test To Prioritize and Evaluate Chemical Biodegradability.

Comprehensive assessment of environmental biodegradability of pollutants is limited by the use of low throughput systems. These are epitomized by the Organisation for Economic Cooperation and Development (OECD) Ready Biodegradability Tests (RBTs), where one sample from an environment may be used to assess a chemical's ability to readily biodegrade or persist universally in that environment. This neglects the considerable spatial and temporal microbial variation inherent in any environment. Inaccurate designations of biodegradability or persistence can occur as a result. RBTs are central in assessing the biodegradation fate of chemicals and inferring exposure concentrations in environmental risk assessments. We developed a colorimetric assay for the reliable quantification of suitable aromatic compounds in a high throughput biodegradation screening test (HT-BST). The HT-BST accurately differentiated and prioritized a range of structurally diverse aromatic compounds on the basis of their assigned relative biodegradabilities and quantitative structure-activity relationship (QSAR) model outputs. Approximately 20 000 individual biodegradation tests were performed, returning analogous results to conventional RBTs. The effect of substituent group structure and position on biodegradation potential demonstrated a significant correlation (P < 0.05) with Hammett's constant for substituents on position 3 of the phenol ring. The HT-BST may facilitate the rapid screening of 100 000 chemicals reportedly manufactured in Europe and reduce the need for higher-tier fate and effects tests.

'I think you just learnt as you went along' - community clergy's experiences of and attitudes towards caring for dying people: A pilot study.

BACKGROUND: Spiritual distress is a factor associated with poor outcomes at the end of life. Timely interventions, assessing and meeting spiritual distress, among patients are contained within nationally agreed guidance. Community clergy are well placed to work alongside healthcare professionals and chaplains to meet spiritual needs. METHODS: Qualitative interviews among Christian clergy in two South East London boroughs and a self-completed Death Anxiety Questionnaire. RESULTS: Fourteen clergy were interviewed from six Christian denominations. Participants described their experiences of ordination training and how helpful this had been for their work among Christian communities. Respondents were invited to discuss their knowledge of and involvement with palliative care services. Each interviewee also accounted for their understanding of pastoral care and spiritual care and considered whether any differences existed between these terms and, if so, what they were. Overall, clergy lacked any detailed formal training and had little experience of working with or relating to palliative care providers. Recommendations are made to improve educational opportunities and working relationships. CONCLUSIONS: Creating opportunities for clergy and palliative care staff to meet and undertake shared training will enhance the quality and level of care for people dying at home who wish to receive spiritual support. Enabling clergy to develop links with local palliative care centres will enhance confidence for both clergy and staff.

PA6†Death chat: engaging with dying and death.

BACKGROUND: Talking about death continues to be a social taboo. St Christopher's has a large, welcoming social space, (The Anniversary Centre) and is committed to opening up its buildings in a number of ways. The St Christopher's social programme, of which Death Chat is part, aims to break down social taboos. AIMS: Hospices have a responsibility to engage creatively with patients, family members, carers and the wider community. Death Chat, held in the hospice buildings, enables honest discussion about dying and death and topics surrounding these themes. METHODS: Death Chat meets weekly and is an open meeting that takes a different subject each week as the starting point for conversation. Cheese and wine are shared and participants quickly find a place in the group. RESULTS: Death Chat has attracted patients, family members, bereaved relatives and the community since September 2013. Attendees have reflected that coming has broken taboos. Peter said, 'it's nowhere near as depressing as it sounds; it's a nice, friendly atmosphere - a convivial place.' Molly found Death Chat to be a welcoming, open and challenging space, 'I have learnt that death is more about my attitude to life than anything else. It has been by far the most important lesson I have learnt since dealing with bereavement.' CONCLUSION: Death Chat provides a forum in which discussion of dying and death for recognises that these are social events and reclaims them from being taboo, to being a normal part of life's experience.

Standard inocula preparations reduce the bacterial diversity and reliability of regulatory biodegradation tests.

OECD ready biodegradability tests have been central to understanding the biodegradation of chemicals from a regulatory perspective for many decades. They are not fit for contemporary prioritisation of chemicals based on persistence, however, due to the low concentration of inocula used, short duration and high variability between tests. Two OECD standard inoculum pretreatment methods (settlement and filtration) were investigated to observe their effect on the probability of biodegradation and associated changes in bacterial community structure and diversity of inocula sourced from the activated sludge process of wastewater treatment plants. Both settlement and filtration were shown to dramatically and significantly reduce the probability and increase the variability of biodegradation of 4-nitrophenol compared to the use of unprocessed inocula. These differences were associated with a significant hundred-fold reduction in cell numbers and solids content and a significant shift in bacterial community structure that was sometimes accompanied by significant reductions in detectable operational taxonomic unit richness and evenness. The natural variation (between different environments) and variation due to differential selection of bacterial communities (by different pretreatment methods) is offered as an explanation for the historical high variability in standard OECD ready biodegradability tests.