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Enzyme purification, characterization, and identification are some of the best ways to introduce undergraduate students to many aspects of biochemistry, particularly as part of project-based learning (PBL). These kinds of multi-step laboratory experiments not only help students to better understand basic biochemistry concepts but also serve to introduce them to the scaffolded nature of the research environment. A 13-week enzyme-based laboratory project was designed as one of three components associated with the course titled Capstone Laboratory, which is delivered to second-year undergraduate students at Weill Cornell Medicine in Qatar (WCM-Q). The project incorporated several fundamental biochemical laboratory techniques, such as chromatography, centrifugation, spectrophotometry, electrophoresis, and kinetic assays, as well as enzyme inhibition and bioinformatic exercises. The aims of the project were to first purify, then to quantify, and finally to study a particular lactate dehydrogenase (LDH) isoenzyme extracted from different chicken organs. LDH was selected for investigation because its inhibition has potential as a therapeutic strategy for cancer treatment. Students enrolled in the Capstone Laboratory course were divided into three groups. Each group conducted experiments associated with one of the project's three aims over consecutive 3-week periods. Relevant data and materials were passed from one group to the next, with individual students writing reports describing the results from their respective collection of experiments. Students in the third and final group gave presentations summarizing the results of the overall project. In the associated bioinformatic exercises, students assessed the similarities and differences between chicken-sourced and human-sourced LDH as well as the interaction between the LDH enzyme and the inhibitors. This PBL in biochemistry is a successful addition to the WCM-Q premedical curriculum because (a) it affords the second-year premedical students opportunities to improve and develop content knowledge and technical and communication skills, and also (b) it provides an opportunity to engage many of the undergraduate students in research.

Since COVID‐19 took a strong hold around the globe causing considerable morbidity and mortality, a lot of effort was dedicated to manufacturing effective vaccines against SARS‐CoV‐2. Many questions have since been raised surrounding the safety of the vaccines, and a lot of media attention to certain side effects. This caused a state of vaccine hesitancy that may prove problematic in the global effort to control the virus. This review was undertaken with the aim of putting together all the reported cardiovascular and haematological events post COVID‐19 vaccination in published literature and to suggest possible mechanisms to explain these rare phenomena. [ABSTRACT FROM AUTHOR]

Myeloid cells, including antigen-presenting cells (APCs) and myeloid-derived suppressor cells (MDSCs) play opposing roles to orchestrate innate and adaptive immune responses during physiological and pathological conditions. We investigated the role of DNA methylation in regulating the transcription of inhibitory/suppressive molecules in myeloid suppressive cells (identified as CD33+HLA-DR–) in comparison to APCs. We selected a number of immune checkpoints (ICs), IC ligands, and immunosuppressive molecules that have been implicated in MDSC function, including PD-L1, TIM-3, VISTA, galectin-9, TGF-β, ARG1 and MMP9. We examined their mRNA expression levels, and investigated whether DNA methylation regulates their transcription in sorted myeloid cell subpopulations. We found that mRNA levels of PD-L1, TIM-3, TGF-β, ARG1 and MMP9 in CD33+HLA-DR– cells were higher than APCs. However, VISTA and galectin-9 mRNA levels were relatively similar in both myeloid subpopulations. CpG islands in the promoter regions of TGF-β1, TIM-3 and ARG1 were highly unmethylated in CD33+HLA-DR–cells, compared with APCs, suggesting that DNA methylation is one of the key mechanisms, which regulate their expression. However, we did not find differences in the methylation status of PD-L1 and MMP9 between CD33+HLA-DR– and APCs, suggesting that their transcription could be regulated via other genetic and epigenetic mechanisms. The promoter methylation status of VISTA was relatively similar in both myeloid subpopulations. This study provides novel insights into the epigenetic mechanisms, which control the expression of inhibitory/suppressive molecules in circulating CD33+HLA-DR– cells in a steady-state condition, possibly to maintain immune tolerance and haemostasis. [ABSTRACT FROM AUTHOR]

Myeloid cells, including antigen-presenting cells (APCs) and myeloid-derived suppressor cells (MDSCs) play opposing roles to orchestrate innate and adaptive immune responses during physiological and pathological conditions. We investigated the role of DNA methylation in regulating the transcription of inhibitory/suppressive molecules in myeloid suppressive cells (identified as CD33 + HLA-DR - ) in comparison to APCs. We selected a number of immune checkpoints (ICs), IC ligands, and immunosuppressive molecules that have been implicated in MDSC function, including PD-L1, TIM-3, VISTA, galectin-9, TGF-β, ARG1 and MMP9. We examined their mRNA expression levels, and investigated whether DNA methylation regulates their transcription in sorted myeloid cell subpopulations. We found that mRNA levels of PD-L1, TIM-3, TGF-β, ARG1 and MMP9 in CD33 + HLA-DR - cells were higher than APCs. However, VISTA and galectin-9 mRNA levels were relatively similar in both myeloid subpopulations. CpG islands in the promoter regions of TGF-β1, TIM-3 and ARG1 were highly unmethylated in CD33 + HLA-DR - cells, compared with APCs, suggesting that DNA methylation is one of the key mechanisms, which regulate their expression. However, we did not find differences in the methylation status of PD-L1 and MMP9 between CD33 + HLA-DR - and APCs, suggesting that their transcription could be regulated via other genetic and epigenetic mechanisms. The promoter methylation status of VISTA was relatively similar in both myeloid subpopulations. This study provides novel insights into the epigenetic mechanisms, which control the expression of inhibitory/suppressive molecules in circulating CD33 + HLA-DR - cells in a steady-state condition, possibly to maintain immune tolerance and haemostasis.