Evaluating patient satisfaction with primary care consultations in the Helderberg sub-district of South Africa

Family and Emergency Medicine: Family Medicine and Primary Carehttps://academic.oup.com/fampra/article/36/3/310/5091262?login=tru

Recombinant production of human ICAM-1 chimeras by single step on column 2 refolding and purification

The interaction of the adhesion molecule of the immunoglobulin family Intercellular Adhesion Molecule 1 (ICAM-1) with its ligands such the integrins LFA-1 and Mac-1 is crucial for the regulation of several physiological and pathophysiological processes like cell mediated-elimination of tumor or virus infected cells, cancer metastasis or inflammatory autoimmune processes. Thus, production of milligrams of protein is required to perform structural and functional studies as well as design novel approaches to find out new inhibitors of ICAM-1/LFA-1 interaction. Here we report on the production of a recombinant human ICAM-1 chimera comprising the first two extracellular domains of ICAM-1 linked to the Fc fraction of a human IgG1. To this aim we have used a cost-effective method based on the expression of a His-tagged protein in E. coli followed by a single step of refolding and purification on immobilized metal affinity columns. This method is able to produce 3mg/liter of bacterial culture in just 72 hours with purity greater than 95%. The identity and the native structure of refolded human ICAM-1 chimera were confirmed by biochemical and biophysical studies including SDS-electrophoresis, immunoblot, circular dichroism, isothermal titration calorimetry and fluorescence spectroscopy. Native folding and functional activity of the chimera were further confirmed by different cell biology studies, including B cell adhesion, T cell binding and inhibition of NK cell function. These studies indicate a high biological activity of the protein since it induces a 200-fold increase/mg of protein in B cell adhesion and the Inhibitory Dose 50 to block cell-mediated cytotoxicity is 10 pg/effector cell. These analyses show that our protocol is able to produce a recombinant human ICAM-1 chimera fully active and useful to analyse the biological processes in which ICAM-1/LFA-1 interaction is critically involved.This work was supported by grants 2009tw0034 from the Spanish National Research, Council (CSIC) and The National Science Council from Taiwan (EMG and AC), SAF2011-25390 from Spanish Ministry of Economy and Competitiveness (JP) and BFU2010-19451 from Spanish Ministry of Science and Innovation (AVC). JP and AVC were supported by Aragón I+D (ARAID). DSM is supported by a fellowship from Gobierno de Aragón. We thank Klaus Ebnet for advice regarding experimental models and ICAM-1 cloning and Adriana Aporta for helping in the establishment of the purification protocol.Peer reviewe

Evaluation of patient characteristics, management and outcomes for COVID-19 at district hospitals in the Western Cape, South Africa : descriptive observational study

CITATION: Mash, R. J. et al. 2021. Evaluation of patient characteristics, management and outcomes for COVID-19 at district hospitals in the Western Cape, South Africa : descriptive observational study. BMJ Open, 11:e047016, doi:10.1136/bmjopen-2020-047016.The original publication is available at https://bmjopen.bmj.comENGLISH ABSTRACT: Objectives To describe the characteristics, clinical management and outcomes of patients with COVID-19 at district hospitals. Design A descriptive observational cross-sectional study. Setting District hospitals (4 in metro and 4 in rural health services) in the Western Cape, South Africa. District hospitals were small (<150 beds) and led by family physicians. Participants All patients who presented to the hospitals’ emergency centre and who tested positive for COVID-19 between March and June 2020. Primary and secondary outcome measures Source of referral, presenting symptoms, demographics, comorbidities, clinical assessment and management, laboratory turnaround time, clinical outcomes, factors related to mortality, length of stay and location. Results 1376 patients (73.9% metro, 26.1% rural). Mean age 46.3 years (SD 16.3), 58.5% females. The majority were self-referred (71%) and had comorbidities (67%): hypertension (41%), type 2 diabetes (25%), HIV (14%) and overweight/obesity (19%). Assessment of COVID-19 was mild (49%), moderate (18%) and severe (24%). Test turnaround time (median 3.0 days (IQR 2.0–5.0 days)) was longer than length of stay (median 2.0 day (IQR 2.0–3.0)). The most common treatment was oxygen (41%) and only 0.8% were intubated and ventilated. Overall mortality was 11%. Most were discharged home (60%) and only 9% transferred to higher levels of care. Increasing age (OR 1.06 (95% CI 1.04 to 1.07)), male (OR 2.02 (95% CI 1.37 to 2.98)), overweight/obesity (OR 1.58 (95% CI 1.02 to 2.46)), type 2 diabetes (OR 1.84 (95% CI 1.24 to 2.73)), HIV (OR 3.41 (95% CI 2.06 to 5.65)), chronic kidney disease (OR 5.16 (95% CI 2.82 to 9.43)) were significantly linked with mortality (p<0.05). Pulmonary diseases (tuberculosis (TB), asthma, chronic obstructive pulmonary disease, post-TB structural lung disease) were not associated with increased mortality. Conclusion District hospitals supported primary care and shielded tertiary hospitals. Patients had high levels of comorbidities and similar clinical pictures to that reported elsewhere. Most patients were treated as people under investigation. Mortality was comparable to similar settings and risk factors identified.https://bmjopen.bmj.com/content/bmjopen/11/1/e047016.full.pdfPublisher's versio

The importance of immune dysfunction in determining outcome in acute liver failure

Acute liver failure (ALF) shares striking similarities with septic shock with regard to the features of systemic inflammation, progression to multiple organ dysfunction and functional immunoparesis. While the existence of opposing systemic pro- and anti-inflammatory profiles resulting in organ failure and immune dysfunction are well recognised in septic shock, characterization of these processes in ALF has only recently been described. This review explores the evolution of the systemic inflammation in acute liver failure, its relation to disease progression, exacerbation of liver injury and development of innate immune dysfunction and extra-hepatic organ failure as sequelae. Defects in innate immunity are described in hepatic and extra-hepatic compartments. Clinical studies measuring levels of pro- and anti-inflammatory cytokines and expression of the antigen presentation molecule HLA-DR on monocytes, in combination with ex-vivo experiments, demonstrate that the persistence of a compensatory anti-inflammatory response syndrome, leading to functional monocyte deactivation, is a central event in the evolution of systemic immune dysfunction. Accurate immune profiling in ALF may permit the development of immunomodulatory strategies in order to improve outcome in this condition