Blood donation screening for hepatitis B virus core antibodies: The importance of confirmatory testing and initial implication for rare blood donor groups.

BACKGROUND AND OBJECTIVES: Exclusion of blood donors with hepatitis B virus (HBV) core antibodies (anti-HBc) prevents transfusion-transmitted HBV infection but can lead to significant donor loss. As isolated anti-HBc positivity does not always indicate true past HBV infection, we have investigated the effectiveness of confirmatory anti-HBc testing and the representation of rare blood groups in anti-HBc-positive donors. MATERIALS AND METHODS: Three hundred ninety-seven HBV surface antigen-negative and anti-HBc initially reactive blood donor samples were tested by five different anti-HBc assays. RESULTS: Eighty percentage of samples reactive in Architect anti-HBc assay were positive by the Murex assay and anti-HBc neutralization. Eleven out of 397 samples showed discordant results in supplementary testing from the Murex confirmatory test result, and five remained undetermined following extensive serological testing. Thirty-eight percentage of anti-HBc-positive donors identified as minority ethnic groups compared with 11% representation in anti-HBc-negative donors (p < 0.0001); the frequency of the Ro blood group in anti-HBc-positive donors was 18 times higher in non-white ethnic groups. CONCLUSION: Using two anti-HBc assays effectively enabled the identification of HBV-exposed and potentially infectious donors, their deferral and potential clinical follow-up. However, the exclusion of confirmed anti-HBc-positive donors will still impact the supply of rare blood such as Ro.

Toward the rapid diagnosis of sepsis: dendritic copper nanostructure functionalized diazonium salt modified screen-printed graphene electrode for IL-6 detection.

Sepsis, an infectious disease affecting millions of people's health worldwide each year, calls for urgent attention to an improvement of analytical devices. Chemiluminescence immunoassay is a typical diagnostic method utilized to assess the risk development of sepsis. However, due to its high-cost, delayed, and complicated procedure, the practical utilization is therefore undoubtedly limited, especially for point-of-care test. Herein, we fabricated for the first time an immunosensor based on dendritic copper nanostructures (CuNSs) combined with 4-aminobenzoic acid (4-AB, the diazonium salt) as antibody linker modified on a screen-printed graphene electrode for the early detection of the sepsis biomarker interleukin-6 (IL-6). The electrode fabrication is made by electrodeposition, thus eliminating the multistep of nanomaterial synthesis and time wasting. The resulting dendritic CuNSs significantly increase the effective surface area (1.2 times) and the sensor's performance. The morphology of this combination was characterized using CV, EIS, SEM, EDX, and FTIR techniques. In the detection process, the appearance of IL-6 suppresses the current response of the redox probe indicator measured by differential pulse voltammetry due to the antibody-antigen complex. The subtraction of signal (ΔI) was interpreted as IL-6 concentration. This sensor exhibited a linear range from 0.05 to 500 pg mL-1 with low detection limit of 0.02 pg mL-1, proving a possibility for early sepsis screening. In addition, the established immunosensor can successfully quantify IL-6 in human serum sample, in which the results agreed well with those achieved using the standard approach, further showing high practical applicability of this developed immunosensor.

The Benefits of Using Case Study Focussed, Problem Based Learning Approaches to Unit Design for Biomedical Science Students.

As part of the Biomedical Sciences undergraduate degree course students are required to apply biological principles to the interpretation of clinical case studies and the diagnosis of patients. Case study-based learning, i.e., application of knowledge to patient diagnosis, is new to most students as case studies do not form part of non-applied A level courses in biological sciences. This approach is an example of Problem Based Learning (PBL) which has been shown to support higher levels of student learning, encouraging critical thinking and analysis. PBL approaches have also been shown to increase academic satisfaction and student engagement. In recent years we have observed a downwards trend in student engagement and historically student performance in applied case study-based assessments to be lower than that observed for assessments based on detailing fundamental biological principles. We hypothesised that PBL teaching delivery would support students in preparing for case study-based assessments, helping them to demonstrate their critical evaluation and problem-solving skills, and hence, improve student performance. We also hypothesised that the student learning experience would be enhanced by a PBL teaching delivery approach which would improve overall engagement. We therefore redesigned a second year Biomedical Sciences degree haematology and clinical biochemistry unit: "Blood Science," with a stronger focus on PBL, including case study focussed activities throughout the unit. We subsequently analysed whether this PBL-focussed unit design improved student experience and feedback, student engagement and student confidence for biomedical science undergraduate students. We present here, our teaching strategy and the impact our changes had on student feedback for the 21/22 and 22/23 academic years. Our findings demonstrate that case study-based activities and tutorial PBL exercises, when incorporated into the curriculum design, can improve student experience in the Biomedical Sciences and other biological science undergraduate degree courses.

Cold stored platelets in the management of bleeding: is it about bioenergetics?

When platelet concentrates (PCs) were first introduced in the 1960s as a blood component therapy, they were stored in the cold. As platelet transfusion became more important for the treatment of chemotherapy-induced thrombocytopenia, research into ways to increase supply intensified. During the late 1960s/early 1970s, it was demonstrated through radioactive labeling of platelets that room temperature platelets (RTP) had superior post-transfusion recovery and survival compared with cold-stored platelets (CSP). This led to a universal switch to room temperature storage, despite CSP demonstrating superior hemostatic effectiveness upon being transfused. There has been a global resurgence in studies into CSP over the last two decades, with an increase in the use of PC to treat acute bleeding within hospital and pre-hospital care. CSP demonstrate many benefits over RTP, including longer shelf life, decreased bacterial risk and easier logistics for transport, making PC accessible in areas where they have not previously been, such as the battlefield. In addition, CSP are reported to have greater hemostatic function than RTP and are thus potentially better for the treatment of bleeding. This review describes the history of CSP, the functional and metabolic assays used to assess the platelet storage lesion in PC and the current research, benefits and limitations of CSP. We also discuss whether the application of new technology for studying mitochondrial and glycolytic function in PC could provide enhanced understanding of platelet metabolism during storage and thus contribute to the continued improvements in the manufacturing and storage of PC.

What is the context? To transition into an activated state, platelets require a highly efficient source of energy that is met through the production of ATP ­ this is referred to as "platelet bioenergetics"Platelets can be removed from healthy donors and used to make platelet concentrates for clinical usePlatelet concentrates are used clinically either therapeutically (to halt bleeding) or prophylactically (to prevent bleeding in patients with low platelet counts)They are stored at room temperature (20­24^oC) with constant gentle agitation, in packs that allow gas exchange and have a 7-day shelf life in some jurisdictionsStoring platelets in the cold (2­6^oC) has historically been shown to improve their ability to halt bleedingWhat is new? There is a renewed interest in cold stored platelets for use in actively bleeding patientsThere are benefits to cold-storing platelets over room temperature storageCold stored platelets are licensed in the US and Norway for certain indications for 14 daysWhat is next? Cold stored platelets have the potential to improve logistics of clinical supply of platelets, enable supply of platelet concentrates where access is currently limited, such as pre-hospital care and on the battlefield and provide improved hemostatic effects for bleeding patients.New research measuring the bioenergetic profiles of cold stored platelets could advance understanding of metabolism in cold stored platelets and support decisions on their re-introduction on a wider scale.

CXR-Net: A Multitask Deep Learning Network for Explainable and Accurate Diagnosis of COVID-19 Pneumonia From Chest X-Ray Images.

Accurate and rapid detection of COVID-19 pneumonia is crucial for optimal patient treatment. Chest X-Ray (CXR) is the first-line imaging technique for COVID-19 pneumonia diagnosis as it is fast, cheap and easily accessible. Currently, many deep learning (DL) models have been proposed to detect COVID-19 pneumonia from CXR images. Unfortunately, these deep classifiers lack the transparency in interpreting findings, which may limit their applications in clinical practice. The existing explanation methods produce either too noisy or imprecise results, and hence are unsuitable for diagnostic purposes. In this work, we propose a novel explainable CXR deep neural Network (CXR-Net) for accurate COVID-19 pneumonia detection with an enhanced pixel-level visual explanation using CXR images. An Encoder-Decoder-Encoder architecture is proposed, in which an extra encoder is added after the encoder-decoder structure to ensure the model can be trained on category samples. The method has been evaluated on real world CXR datasets from both public and private sources, including healthy, bacterial pneumonia, viral pneumonia and COVID-19 pneumonia cases. The results demonstrate that the proposed method can achieve a satisfactory accuracy and provide fine-resolution activation maps for visual explanation in the lung disease detection. Compared to current state-of-the-art visual explanation methods, the proposed method can provide more detailed, high-resolution, visual explanation for the classification results. It can be deployed in various computing environments, including cloud, CPU and GPU environments. It has a great potential to be used in clinical practice for COVID-19 pneumonia diagnosis.

Electroanalytical point-of-care detection of gold standard and emerging cardiac biomarkers for stratification and monitoring in intensive care medicine - a review.

Determination of specific cardiac biomarkers (CBs) during the diagnosis and management of adverse cardiovascular events such as acute myocardial infarction (AMI) has become commonplace in emergency department (ED), cardiology and many other ward settings. Cardiac troponins (cTnT and cTnI) and natriuretic peptides (BNP and NT-pro-BNP) are the preferred biomarkers in clinical practice for the diagnostic workup of AMI, acute coronary syndrome (ACS) and other types of myocardial ischaemia and heart failure (HF), while the roles and possible clinical applications of several other potential biomarkers continue to be evaluated and are the subject of several comprehensive reviews. The requirement for rapid, repeated testing of a small number of CBs in ED and cardiology patients has led to the development of point-of-care (PoC) technology to circumvent the need for remote and lengthy testing procedures in the hospital pathology laboratories. Electroanalytical sensing platforms have the potential to meet these requirements. This review aims firstly to reflect on the potential benefits of rapid CB testing in critically ill patients, a very distinct cohort of patients with deranged baseline levels of CBs. We summarise their source and clinical relevance and are the first to report the required analytical ranges for such technology to be of value in this patient cohort. Secondly, we review the current electrochemical approaches, including its sub-variants such as photoelectrochemical and electrochemiluminescence, for the determination  of important CBs highlighting the various strategies used, namely the use of micro- and nanomaterials, to maximise the sensitivities and selectivities of such approaches. Finally, we consider the challenges that must be overcome to allow for the commercialisation of this technology and transition into intensive care medicine.

Analysis of heat-shock protein localisation using flow cytometry.

The use of flow cytometry in heat-shock protein (HSP) research is increasing rapidly due to the high sensitivity and versatility of the technique. The method allows the simultaneous analysis of multiple proteins within numerous cell types in a heterogeneous sample, providing advantages over alternative techniques, such as ELISA and Western blotting. As a result, flow cytometry is becoming the leading technique used in this area of research. The current chapter describes the methodology for preparing samples for this technique and outlines two protocols for the analysis of surface- and intracellular-localised HSPs.

Heat Shock Protein translocation induced by membrane fluidization increases tumor-cell sensitivity to chemotherapeutic drugs.

Treatment of chronic lymphocytic leukemia (CLL) remains a challenge due to the frequency of drug resistance amongst patients. Improving the delivery of chemotherapeutic agents while reducing the expression of anti-apoptotic Heat Shock Proteins (HSPs) within the cancer cells may facilitate in overcoming this drug resistance. We demonstrate for the first time that sub-lethal doses of chemotherapeutic agents can be combined with membrane fluidizing treatments to produce a significant increase in drug efficacy and apoptosis in vitro. We show that fluidizers result in a transient decrease in intracellular HSPs, resulting in increased tumor-cell sensitivity and a membrane-associated induction of HSP gene expression.

Differential heat shock protein localization in chronic lymphocytic leukemia.

Mechanisms behind carcinogenesis and resistance of tumor cells to treatment regimes remain elusive. The major stress proteins Hsp72, Hsp90, and Hsp27 are credible candidates to provide this resistance, as their overexpression in many cancer types is well documented. In addition to being present inside tumor cells, where they confer resistance to apoptosis, Hsp72, in particular, is presented externally, embedded in the cell membrane of cancer cells. This study aimed to investigate the localization of Hsp72, Hsp90, and Hsp27 in leukocytes from patients with CLL and age-matched control subjects. CLL patients were found to express significantly higher levels of iHsp90 (CLL=2463 MFI; control=748 MFI) and iHsp27 (CLL=2190 MFI; control=1031 MFI) in lymphocytes than that expressed by lymphocytes from control subjects. Furthermore, expression of iHsp90 was shown to be related to stage of disease, and expression of iHsp27 correlated with levels of active caspase-3. Patients were found to express very high levels or very low levels of sHsp72 and iHsp72 in CD5(+)/CD19(+) cells, although surface and intracellular datasets did not correlate. Levels of extracellular Hsp72 circulating in the serum were found to correlate with internal levels of Hsp72 and were also found to be significantly lower in patients receiving corticosteroid treatment than in patients not receiving corticosteroid treatment. Finally, analysis of the number of circulating Tregs revealed significantly elevated numbers in CLL patients compared with control subjects.