Is drug interference still an issue for pretransfusion testing of patients on anti CD38 and other monoclonal antibody therapies?

Certain therapies that target CD markers on some blood cells can affect pretransfusion testing. Key examples are anti-CD38, CD47 monoclonal antibody (mAb) therapies such as daratumumab (DARA) and magrolimab, which have presented a challenge for transfusion medicine laboratories around the globe. Scientists have been faced with not only introducing a protocol to provide safe blood to patients but also investigating the most effective method to remove the pretransfusion pan-agglutinating interference caused. A number of papers in the last 5 years have reported on various methods to remove pretransfusion interference; however, most of these studies have been conducted only in a few countries. Most recent reviews on this topic have focused on techniques and reagents to remove pretransfusion interference, and dithiothreitol is currently the gold standard for removing DARA interference. However, it was clear from this review that while many laboratories have developed processes for addressing interference in pretransfusion testing, and DARA interference may not be a major issue, there are still laboratories around the world, that may not have adequately addressed this issue. In addition, the impact of mAb interference on widely used techniques such as flow cytometry is unclear.

An Integrated Approach to Teaching Cell Staining.

Active learning has been shown to improve student outcomes across a range of science, technology, engineering, and math (STEM) disciplines. In addition, active learning with an interdisciplinary focus in the classroom is beneficial for students to pursue health and allied health careers. Case studies have also been shown to enhance student learning. In this study, we utilized a novel learning approach combining a case study with interdisciplinary focus and an active learning exercise that introduced the chemistry of stains and how this relates to staining various organelles and components in a cell. This case study along with active learning exercise (e.g., a card game, group PowerPoint presentations, or coloring organelles or structures in a eukaryotic cell upon staining with a biochemical stain) resulted in improved student performance as well as long-term retention. This activity can be implemented in introductory biology, microbiology, cell biology, and related STEM disciplines (e.g., chemistry, biochemistry).

Immune Literacy: a Call to Action for a System-Level Change.

Immune literacy-the ability to hear, learn, read, write, explain, and discuss immunological content with varied audiences-has become critically important in recent years. Yet, with its complex terminology and discipline-specific concepts, educating individuals about the immune system and its role in health and disease may seem daunting. Here, we reflect on how to demystify the discipline and increase its accessibility for a broader audience. To address this, a working group of immunology educators from diverse institutions associated with the research coordination network, ImmunoReach, convened virtually. As a result of these discussions, we request a call to action for a system-level change and present a set of practical recommendations that novice and experienced educators from diverse institutions, professional societies, and policymakers may adopt to foster immune literacy in their classrooms and communities.

Exploring the uptake of COVID-19 vaccination amongst respiratory therapists in Canada.

Introduction/background: COVID-19 vaccination uptake rates and responses by Canadian respiratory therapists (RTs) were investigated along with factors that may be shown to play a role in vaccination hesitancy. Methods: An anonymous survey using SurveyMonkey® on vaccination uptake rates, responses and attitudes was made available to student RTs, graduate RTs and registered RTs in Canada from July to October of 2021. Pearson's chi-square tests were performed to evaluate association between vaccination status and the other categorical parameters evaluated. Results: A total of 1013 surveys (8.0% of target population) were completed fully and included in the data analysis. Of the surveyed RT population, 90.42% received their vaccination as soon as it was made available compared to Canada's Ministry of Health's published rate at the time of 86.27% for all Canadian healthcare workers. There was a significant (p = 0.013) association between early vaccination and age and a significant (p = 0.036) association between vaccination status and a participant's response on whether or not they have a family member or know someone who has had COVID-19. There was also a significant (p < 0.001) association between vaccination status and attitudes towards trusting science to develop safe, effective, new vaccines and trusting the Ministry of Health to ensure that vaccines are safe. There was no significant association between vaccination status and gender, province/territory of residency/work, level of education and level of involvement with COVID-19 patients. Conclusion: The results suggest that RT groups across Canada had higher early vaccination uptake rates than general healthcare worker groups and that age, relationship to people with COVID-19 and trust in science played a significant role in their vaccination uptake rates.

Metagenomics of Toenail Onychomycosis in Three Victorian Regions of Australia.

Onychomycosis is a fungal disease of the nail that is found worldwide and is difficult to diagnose accurately. This study used metagenomics to investigate the microbiology of 18 clinically diagnosed mycotic nails and two normal nails for fungi and bacteria using the ITS2 and 16S loci. Four mycotic nails were from Bass Coast, six from Melbourne Metropolitan and eight from Shepparton, Victoria, Australia. The mycotic nails were photographed and metagenomically analysed. The ITS2 sequences for T. rubrum and T. interdigitale/mentagrophytes averaged over 90% of hits in 14/18 nails. The high abundance of sequences of a single dermatophyte, compared to all other fungi in a single nail, made it the most likely infecting agents (MLIA). Trichophyton rubrum and T. interdigitale/mentagrophytes were found in Bass Coast and Shepparton while only T. interdigitale/mentagrophytes was found in Melbourne. Two nails with T. interdigitale/mentagrophytes mixed with high abundance non-dermatophyte moulds (NDMs) (Aspergillus versicolor, Acremonium sclerotigenum) were also observed. The two control nails contained chiefly Fusarium oxysporum and Malassezia slooffiae. For bacteria, Staphylococcus epidermidis was in every nail and was the most abundant, including the control nails, with an overall mean rate of 66.01%. Rothia koreensis, Corynebacterium tuberculostearicum, and Brevibacterium sediminis also featured.

Antigen and Immunogen: An Investigation into the Heterogeneity of Immunology Terminology in Learning Resources.

The need to focus on immunology education has never been greater. The coronavirus disease 2019 pandemic has revealed that a significant proportion of our society is vaccine hesitant. Some of this hesitancy may stem from a general lack of understanding of how the immune system and immunological interventions work. In addition, social media platforms undercut public health efforts by quickly propagating a multitude of misconceptions and erroneous information surrounding the science behind these interventions. The responsibility to be advocates for science is well recognized by immunology researchers, educators, and public health professionals, as evidenced by the rich body of resources developed to communicate science to the lay audience. Scientific jargon, however, can be a barrier to effective communication and can negatively impact learning and comprehension. The field of immunology is especially laden with discipline-specific terminology, which can hamper educators' efforts to convey key concepts to learners. Furthermore, a lack of consistency in accepted definitions can complicate students' conceptual understanding. Learning resources, including textbooks, published in print or available online, and exclusively digital resources, continue to serve as the primary sources of information for both educators and students. In this article, we describe a vast heterogeneity in learning resource glossary descriptions of two key conceptual terms: antigen and immunogen We provide a perspective on pedagogical strategies to address these critical terms. Using current knowledge, we recommend an approach to standardize the definitions of the terms antigen and immunogen within the immunology educator community.

Challenges in antibody titration for ABO-incompatible renal transplantation.

BACKGROUND AND OBJECTIVES: Accurate and regular monitoring of anti-A and anti-B titres pre- and post-transplantation plays a crucial role in the clinical management of patients receiving ABO-incompatible renal transplants. There is no standardized protocol or an external quality assurance program (EQA) currently available for this testing in Australia. The aim of this study was to investigate the diversity of techniques, test platforms and reagents that were currently in use in various laboratories with the aim of developing an EQA. MATERIALS AND METHODS: An online survey was sent to the participants enrolled with the Royal College of Pathologists of Australasia Quality Assurance Program (RCPAQAP) to assess their interest in participation in the pilot study. A total of 24 participants who expressed interest were sent the group O plasma, A1 , A2 and B cells to perform ABO titration using their own methods. RESULTS: Participants reported a wide range of titre results, from 8 to 1024 for the anti-A titre using A1 cells, from 2 to 128 for anti-A titre using A2 cells and from neat to 32 for anti-B titre using B cells. CONCLUSION: There was a wide variation in titre results between and within different technologies. These findings demonstrate the need for an ABO titration EQA. Development of a standard technique and participation in an EQA program should, over time, reduce variation and enable transferrable results across testing centres, which will assist in consistent clinical interpretation and better outcomes for patients.

Living with food allergy: What this means for children.

Health-related knowledge, skills and attitudes that translate to behaviours are important foundations for healthy living. However, deficiencies in any one of these factors have the potential to impact quality of life. This study investigates how Australian children perceive and manage their food allergy as they transition through childhood onto adolescence. To measure this, children aged 12 years and under completed an online survey [under 8 years and 8-12 years] on knowledge, skills and attitudes. Although recruitment involved advertisements to over 700 Australian pre-schools, 44 allergy specialists and multiple representations to the patient support group, Allergy & Anaphylaxis Australia, completion relied on parental notification and/or guidance. Of the 139 participants recruited, 114 commenced the survey with 65 participants under 8 years (58%) and 49 between 8 and 12 years. Results showed participants recognised different ways to keep safe with children under 8 years (N = 65) 100% cognisant of the need to check with 'Mum/Dad' or teachers if they could safely eat food as well as the importance of not swapping lunches, while there was a slight change in compliance for those aged 8-12 years as three (6.1%, n = 3/49) children did not see the necessity of asking an adult/teacher if a food item was safe to eat. Older children were also less likely to tell friends about their food allergy even if they thought they were having an allergic reaction, despite children under 8 years more likely to feel different to friends (38%, n = 24/64) compared to those aged 8-12 years (31%, n = 14/45). Although both groups disliked parents' fussing, for children under 8 years, there was a strong association between this and 'feeling different to friends'. Results from this study highlight the importance of normalising food allergy at home and in the community, so children are confident in telling others about their food allergy.

A Systematic Review of Campylobacter jejuni Vaccine Candidates for Chickens.

Campylobacter jejuni infection linked to the consumption of contaminated poultry products is one of the leading causes of human enteric illness worldwide. Vaccination of chickens is one of the potential strategies that could be used to control C. jejuni colonization. To date, various C. jejuni vaccines using potential antigens have been evaluated, but a challenge in identifying the most effective formulation is the wide variability in vaccine efficacies reported. A systematic review was undertaken to compare C. jejuni vaccine studies. Based upon specific selection criteria eligible papers were identified and included in the analysis. Vaccine efficacy reported from different C. jejuni antigens, vaccine types, and vaccination regimens reported in these papers were reviewed. Our analysis shows that total outer membrane proteins and cysteine ABC transporter substrate-binding protein were among the most efficacious vaccine antigen candidates reported. This review also highlights the importance of the need for increased consistency in the way C. jejuni vaccine studies in poultry are designed and reported in order to be able to undertake a robust comparison of C. jejuni vaccine candidates.

Inside the Undergraduate Immunology Classroom: Current Practices that Provide a Framework for Curriculum Consensus.

Although immunological research has become increasingly important in recent decades for understanding infectious and immune-mediated diseases, immunological pedagogy at the undergraduate level has lagged behind in reports of evidence-based scholarship. To address the need for a renewed emphasis on immunology education and to describe the current status of undergraduate education in immunology, an online survey of instructors with experience in teaching immunology was conducted. The survey investigated the effects of instructors' level of teaching experience, target student population, and course components on the emphasis given to certain immunology subtopics in their courses. Instructor teaching experience and current role in teaching influenced the proportion of time allotted to lab techniques, clinical topics, and evolutionary aspects, but type of institution (undergraduate and graduate degree-granting institutions) did not affect course content or emphasis on subtopics. Topics that received the greatest emphasis were the adaptive immune system, the innate immune system, host-pathogen interactions, and molecular mechanisms. Vaccines, hypersensitivity, autoimmunity, and essential immunology techniques were ranked slightly lower, while topics such as evolution, metabolism and antibody purification received the least emphasis. Inclusion of a lab component increased time given to lab-related and clinical topics but did not affect the perceived importance of various scientific competencies. These data describe current curricular practices of instructors who have experience teaching immunology and inform curricular priorities and course design frameworks for undergraduate immunology education.

Investigation of Campylobacter colonization in three Australian commercial free-range broiler farms.

Campylobacter spp. contaminated poultry products are strongly associated with foodborne illnesses worldwide. Development of effective management strategies to reduce contamination by Campylobacter spp. requires an improved understanding of the numerous factors that drive these contamination processes. Currently, chicken farms are using more free-range chicken meat production systems in response to consumer preferences. However, Campylobacter spp. colonization has rarely been investigated on free-range broiler farms. The present study investigated the temporal and environmental factors influencing Campylobacter spp. colonization of free-range broilers as well as potential sources and genetic diversity of Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli) in commercial free-range broiler farms. Genetic linkages among the isolates were analyzed using flaA amplicon analysis. Campylobacter coli was first detected in fecal samples of a commercial free-range broiler flock on day 10 of rearing. Multiple genotypes of C. jejuni and C. coli were identified in this study. The farm environment was identified as a potential source of C. jejuni and C. coli colonization of free-range broilers. The dominant Campylobacter genotype varied between free-range broiler farms over time, with C. jejuni being the most frequently isolated species. These findings enhance the understanding of C. jejuni and C. coli colonization in free-range broiler farms and could inform the development of more effective intervention strategies to help control this important foodborne pathogen.

Parent perceptions in managing children with food allergy: An Australian perspective.

INTRODUCTION: Food allergy affects up to 10% of Australian children, and living with food allergic children can be challenging for parents. This study explored parental perceptions and knowledge as they navigate a new reality of keeping their child safe. METHODS: Parents of children with food allergies completed an online food allergy survey in 2015. Questions explored health knowledge, skills, and attitudes (KSAs) as well as quality of life (QoL) through the inclusion of the Food Allergy Quality of Life - Parental Burden instrument (FAQL-PB). Notification of the survey included advertisements to more than 700 randomly selected Australia-wide preschools, 44 allergy specialists, and Allergy & Anaphylaxis Australia. Responses were tabulated and analysed. RESULTS: Of the 400 participants who logged on, 357 commenced the survey and 318 finished. Questionnaire analysis showed that 44.1% of parents (n = 135) hesitated to use an adrenaline auto-injector and may be influenced by a classification system where symptom severity is not universally understood. While 79% would sign up to a national Anaphylaxis Registry, intention to participate in clinical trials using vaccines was disclosed by only 56%. Allergen labelling and community acceptance continue to be the most challenging aspects of managing a food allergy, and 50% of parents reported that food allergy played a role in choosing a preschool or primary school. Overall, quality of life for participants was influenced more by gender (male) and age of the child than where they lived - capital city or regional location; however, regional participants were more frustrated over lack of appreciation relating to the seriousness of food allergy (p = 0.010). CONCLUSION: Results highlight the need for educational strategies for both the food allergy community and public, particularly in regional areas, since there is a perceived lack of appreciation as to the seriousness of food allergy. Educational resources and relevant networks are required to support parents and caregivers in the management of children with food allergy.

Predominance of Trichophyton interdigitale Revealed in Podiatric Nail Dust Collections in Eastern Australia.

Toenail onychomycosis caused by dermatophytes is a significant medical and financial worldwide burden. Relatively scant research has been undertaken as to the predominant species and strains causing this condition in Australia, which is a unique isolated continent with diverse geographical, climatic and population regions. Four regions were selected in Eastern Australia: Far North Queensland, Rural Victoria, Melbourne Metropolitan and Tasmania. From each of these areas, communal nail dust bags from podiatric physicians' work were collected and analysed. A total of 32 dust bags were collected: 10 from Far North Queensland, 8 from Melbourne Metropolitan, 8 from Rural Victoria and 6 from Tasmania. Dermatophyte test medium was used to isolate dermatophytes from the dust, and the colonies were subcultured to Potato Dextrose Agar. Of the bags collected, in total 69% were positive for dermatophytes: 40% from Far North Queensland, 75% from Melbourne Metropolitan, 88% from Rural Victoria and 83% from Tasmania. The internal transcribed spacer (ITS) region of ribosomal DNA was used to identify and compare isolates. A total of 148 dermatophyte strains were identified. The predominant species isolated was Trichophyton interdigitale (125 isolates), which was found in all four regions. This species was further subdivided into four ITS genotypes: the first two were present in all regions, but the third was found only in the Melbourne Metropolitan area and the fourth only in Tasmania. Only one strain of Trichophyton rubrum was found and only in Rural Victoria. Eighteen isolates of Arthroderma quadrifidum were cultured from Rural Victoria and Tasmania and were further classified into three ITS genotypes. Some isolates rarely reported in clinical material were identified as Paraphyton cookei, Arthroderma tuberculatum and Arthroderma crocatum. A potentially new species of Arthroderma was also found in Melbourne Metropolitan. These findings reveal a unique dermatophyte fingerprint in toenails for Eastern Australia.

Semi-automated von Willebrand factor multimer assay for von Willebrand disease: Further validation, benefits and limitations.

INTRODUCTION: Accurate diagnosis of von Willebrand disease (VWD) enables effective patient management. von Willebrand factor (VWF) multimer analysis provides useful information regarding VWF multimer structure, thereby aiding VWD subtyping and management; however, historically technically challenging assays have had limited utility. This study evaluates the Sebia Hydrasys Hydragel-11 semi-automated VWF multimer assay and further validates the Hydragel-5 gel system, as primarily pertaining to VWD diagnostics and monitoring of therapy. METHODS: Provisionally diagnosed (via a reference assay test panel) archived patient samples and prospective test patient samples, including those undergoing desmopressin trial or therapy monitoring, along with commercial and in-house control material and various external quality assessment (EQA) samples, were analysed. VWF multimers were evaluated for presence, loss or partial loss of high molecular weight (HMWM) and intermediate molecular weight (IMWM) multimers by both visual inspection and densitometric scanning, and comparison with reference assay results. RESULTS: All anticipated multimer patterns were reproduced, with patients generally showing multimer profiles matching expected patterns according to VWD type based on reference test panel 'diagnosis'. Occasional discrepancies were resolved by retesting. The increase in plasma VWF following desmopressin therapy was also clearly demonstrated. Multimer profiles of EQA samples complemented reference test panel results and matched EQA targets. There were some 'technical' limitations noted. CONCLUSION: This easy to use, standardised, semi-automated multimer analysis system can demonstrate the multimer profile of VWD patients, thus representing an additional laboratory tool for improved diagnosis, thereby facilitating appropriate patient management.

Technologies for the Selection, Culture and Metabolic Profiling of Unique Rhizosphere Microorganisms for Natural Product Discovery.

Small molecule discovery has benefitted from the development of technologies that have aided in the culture and identification of soil microorganisms and the subsequent analysis of their respective metabolomes. We report herein on the use of both culture dependent and independent approaches for evaluation of soil microbial diversity in the rhizosphere of canola, a crop known to support a diverse microbiome, including plant growth promoting rhizobacteria. Initial screening of rhizosphere soils showed that microbial diversity, particularly bacterial, was greatest at crop maturity; therefore organismal recovery was attempted with soil collected at canola harvest. Two standard media (Mueller Hinton and gellan gum) were evaluated following inoculation with soil aqueous suspensions and compared with a novel "rhizochip" prototype buried in a living canola crop rhizosphere for microbial culture in situ. Following successful recovery and identification of 375 rhizosphere microbiota of interest from all culture methods, isolates were identified by Sanger sequencing and/or characterization using morphological and biochemical traits. Three bacterial isolates of interest were randomly selected as case studies for intensive metabolic profiling. After successful culture in liquid media and solvent extraction, individual extracts were subjected to evaluation by UHPLC-DAD-QToF-MS, resulting in the rapid characterization of metabolites of interest from cultures of two isolates. After evaluation of key molecular features, unique or unusual bacterial metabolites were annotated and are reported herein.

Antimicrobial susceptibility of clinical isolates of Campylobacter jejuni from New South Wales, Australia.

OBJECTIVES: The aim of this study was to investigate the prevalence of resistance to commonly used antimicrobials in Campylobacter jejuni isolates from clinical faecal samples in New South Wales (NSW), Australia. METHODS: A total of 117 C. jejuni isolates from human faecal samples from regional and metropolitan NSW were examined for antimicrobial resistance. RESULTS: Of the 117 isolates tested, 15.4% were resistant to ampicillin, 5.1% to tetracycline and 13.7% to ciprofloxacin. Most of the isolates were susceptible to erythromycin, except for three that showed intermediate resistance. Furthermore, 9.4% of isolates were resistant (or intermediate-resistant) to more than one antimicrobial agent. Isolates that were resistant to ampicillin and tetracycline harboured the blaOXA-61 and tet(O) genes, respectively. A mutation in the gyrA gene, resulting in the T86I substitution, was identified in the majority of ciprofloxacin-resistant isolates. CONCLUSION: The data obtained in the current study demonstrate that the majority of C. jejuni isolates evaluated were susceptible to one or more antimicrobials tested. Apart from three isolates that demonstrated intermediate resistance, all of the isolates were susceptible to erythromycin, which is the drug of choice for treating Campylobacter infections.

Chemopreventive Potential of Cereal Polyphenols.

It has been identified that diet is one of the major contributing factors associated with the development of cancer and other chronic pathologies. In the recent years, supplementing regular diet with food and/or its components that contain chemopreventive properties has been considered an effective approach in reducing the incidence of cancer and other lifestyle associated diseases. This systematic review provides an exhaustive summary of the chemopreventive properties exhibited by everyday dietary ingredients such as rice, barley, oats, and sorghum. The studies both in vitro and in vivo reviewed have highlighted the potential role of their polyphenolic content as chemopreventive agents. Polyphenolic compounds including anthocyanins, tricin, protocatechualdehyde, avenanthramide, and 3-deoxyanthocyanins found in rice, barley, oats, and sorghum, respectively, were identified as compounds with potent bioactivity. Studies demonstrated that cereal polyphenols are likely to have chemopreventive activities, particularly those found in pigmented varieties. In conclusion, findings suggest that the consumption of pigmented cereals could potentially have an important role as a natural complementary cancer preventive therapeutic. However, further studies to develop a complete understanding of the mechanisms by which phenolic compounds inhibit cancerous cell proliferation are warranted.

B-Cell Epitope Mapping Using a Library of Overlapping Synthetic Peptides in an Enzyme-Linked Immunosorbent Assay.

This chapter describes a strategy for mapping linear B-cell epitopes of proteins using synthetic biotinylated peptides in an ELISA.A set of overlapping peptides were designed based upon a known amino acid sequence of the target protein, VapA (Virulence-associated Protein A) of the bacterium Rhodococcus equi, an important pulmonary pathogen in foals. The peptides synthesized as biotinylated peptides were coated directly onto micro titer plates which had been pre-coated with NeutrAvidin™ and used to screen sera from foals confirmed to have R. equi disease. A linear B-cell epitope was identified which corresponded to a 20 mer sequence of the VapA protein.

Long-Term Nutritional Outcome and Health Related Quality of Life of Patients Following Esophageal Cancer Surgery: A Meta-Analysis.

Long term health related quality of life (HRQL) and nutritional outcome of patients following esophagectomy for cancer has become increasingly significant as the 5-year survival rate in this patient group is increasing. This meta-analysis aims to investigate the HRQOL, nutritional impact symptoms and nutritional outcomes of patients following an esophagectomy at greater than 12 months after surgery. In studies reporting on HRQL as an outcome, global QOL score at 6-month compare to greater than 12-month showed no statistically significant difference (65.92 vs. 75.78, p = 0.07). Forty-one percent of patients reported a greater than 10% weight loss at six-month follow-up (95% CI: 20-65%; I2 = 94.27, p < 0.001), and at the greater than 12-month follow-up, 33% of patients had the greater than 10% weight loss (95% CI: 15-57%; I2 = 96.18, p < 0.001). At the 12-month or longer post esophagectomy, just over half the patients reported dysphagia (51%, 95% CI: 25-76%; I2 = 95.70, p < 0.001), nausea was reported by 11% (95% CI: 7-19%; I2 = 59.31, p = 0.09), dumping syndrome reported by 60% (95% CI: 43-76%; I2 = 96.92, p < 0.001). Symptoms such as dysphagia, diarrhea, reflux, dumping syndrome, and nausea were found to persist following esophagectomy. There were insufficient robust research investigating how these symptoms impact on the adequacy of dietary intake and micronutrient status.