Production of α-Bisabolol from metabolically engineered Escherichia coli

α-Bisabolol is a natural-occurring sesquiterpenoid with applications in cosmetics as whitening and soothing agent. It is synthesized from the universal precursors, isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP), which are generated either through the mevalonate (MVA) pathway or the 2C-methyl-D-erythritol-4-phosphate (MEP) pathway. Farnesyl pyrophosphate (FPP) synthase (IspA) then catalyzes the condensation of IPP and DMAPP to the linear FPP, which is rearranged and cyclized to α-bisabolol by bisabolol synthases. Here, we compared the capacity of 5 α-bisabolol synthases from Lippia dulcis, Streptomyces citricolor, Santalum spicatum, Matricaria recutita, and Artemisia annua for α-bisabolol production. MVA pathway and FPP synthase were also overexpressed to supply sufficient FPP for bisabolol synthesis in the recombinant E. coli. Bisabolol synthase from M. recutita (MrBBS) shows the highest activity of bisabolol synthesis, and 75 mg/L/OD600 of bisabolol was produced in a test-tube culture. We further optimized the expression level of IspA and MrBBS by modulation their RBS strength. The 24 bisabolol synthesis operons with different RBSs were assessed for their performance on bisabolol synthesis. By this approach, the best strain is able to produce bisabolol with a capacity of 220mg/L/OD600 in a test tube culture. The consequence of host strain optimization led to an increase in bisabolol production to 300 mg/L/OD600, which presents a 4-fold increase over the initial engineered strain. This work was supported by a grant (NRF-2016R1A2B2010678) from the National Research Foundation, MSIP, Korea

Routine breast milk monitoring using automated molecular assay system reduced postnatal CMV infection in preterm infants

Human cytomegalovirus (CMV) transmitted through breast milk poses fatal risks to preterm infants. However, current molecular assay systems often do not accommodate breast milk samples. In this study, we evaluated the analytical and clinical performance of the measurement procedure of CMV load in breast milk utilizing the Cobas CMV test on the Cobas 6,800 system. This was enabled by incorporating a simple independent sample preparation procedure before the application of samples on the automated assay system. Clinical data from electronic medical records were retrospectively analyzed. Breast milk samples from mothers of preterm infants born before 33 weeks of gestation were screened for CMV using the automated assay system. CMV positivity rates in breast milk and neonatal samples and the CMV transmission rate were calculated. Furthermore, to validate the analytical accuracy of the overall measurement procedure with newly obtained residual breast milk samples, the linearity of the measurement procedure was assessed, and a simplified sample preparation method was validated against a conventional method. The CMV positivity rates in maternal breast milk and neonatal samples were 57.8 and 5.2%, respectively. The CMV transmission rate through breast milk was 7.7%. No significant differences in gestational age or birth weight were found between the CMV-negative and CMV-positive neonates. The linearity of the procedure was observed within a range of 1.87–4.73 log IU/mL. The simplified sample preparation method had an equivalent or even improved CMV detection sensitivity than the conventional method. Incorporating a simple independent sample preparation procedure effectively resolved any potential issues regarding the application of breast milk on the automated assay system. Our approach contributed to reduced vertical transmission of CMV by providing a convenient and reliable method for the monitoring of breast milk CMV positivity for clinicians

Enhancing pest control interventions by linking species distribution model prediction and population density assessment of pine wilt disease vectors in South Korea

Pine wilt disease caused by pinewood nematode is one of the most destructive forest diseases, and still spreading in South Korea despite the various control efforts. Japanese pine sawyer (JPS) and Sakhalin pine sawyer (SPS) are the main vectors of the disease. Understanding the distribution and density of the vectors is crucial since the control period is determined by the different emergence periods of the two vectors and the control method by its density and the expected damage severity. In this study, we predicted the distribution of JPS and SPS using Maxent and investigated the relationship between the resulting suitability value and the density. The population densities of JPS and SPS were obtained through a national survey using pheromone traps between 2020-2022. We converted the density data into presence/absence points to externally validate each species distribution model, then we used quantile regression to check the correlation between the suitability and population density, and finally we used three widely used thresholds to convert the model results into binary maps, and tested if they could distinguish the density by comparing the Rb value of biserial correlation. The quantile regression revealed a positive relationship between the habitat suitability and population density sampled in the field. Moreover, the binary map with threshold criteria that maximizes the sum of the sensitivity and specificity had the best density discrimination capacity with the highest Rb. A quantitative relationship between suitability and vector density measured in the field from our study provides reliability to species distribution model as practical tools for forest pest management

Institutional Board Review for Clinical Investigations on Inflammatory Bowel Diseases: A Single-Center Study

Background/AimsThe growing volume and the diversity of clinical research has led to related laws and regulations as well as the Institutional Review Board (IRB) approval process becoming more stringent. To conduct clinical research efficiently and while following regulations, information about the IRB approval process and feedback is important for investigators. This has yet to be studied.MethodsWe included 381 gastrointestinal disease research proposals (79 with inflammatory bowel disease [IBD], and 302 with non-IBD) reviewed by the IRB of Severance Hospital between January 2009 and December 2013. We retrospectively analyzed research characteristics including research risk levels, results of initial reviews, frequencies of continuing review, numbers of IRB comments, frequencies of IRB comments, and durations from submission to approval.ResultsInvestigators' decisions on risk level were higher in the IBD group than in the non-IBD group (P<0.05). Results of initial reviews, frequencies of continuing reviews, the numbers of IRB review comments, and durations from submission to approval were not different between the two groups, but IRB decisions on risk level were higher in the IBD group (P<0.05). In subgroup analysis, the number of IRB comments from initial review on informed consent forms and procedures as well were quest of more information were significantly higher in the IBD group than in the non-IBD group (P<0.001 and 0.01, respectively).ConclusionsIn Korea, rare diseases such as IBD require more information for the IRB process due to their distinct characteristics. IBD researchers should develop research protocols more carefully and make their research as subject-friendly as possible

Propensity score matched analysis for the safety and effectiveness of remdesivir in COVID-19 patients with renal impairment

Backgrounds Remdesivir (RDV) is an antiviral agent approved for the treatment of coronavirus disease 2019 (COVID-19); however, is not recommended for patients with renal impairment. Due to limitations associated with prospective clinical trials, real-world data on the safety and efficacy of RDV in patients with renal impairment are necessary. Methods Propensity score-matched (PSM) retrospective analysis was conducted between March 2020 and September 2022 in COVID-19 patients with an eGFR < 30 mL/min in four Korean hospitals. The RDV treatment group was matched to the untreated control group. The safety and clinical outcomes in patients who received RDV were analyzed. Results A total of 564 patients were enrolled; 229 patients received RDV either for treatment or prophylaxis. On day 5, no difference in nephrotoxicity was observed between the two groups, and liver enzyme levels were within the normal range. In multivariate analysis for new dialysis, RDV treatment was not a risk factor for new dialysis. Among the 564 patients, 417 were indicated for a 5-day course of RDV treatment and 211 patients were treated with RDV. After PSM, no differences in the clinical outcomes were observed between the two groups. Conclusion RDV use in COVID-19 patients with renal impairment did not result in significant nephrotoxicity or hepatotoxicity.This work was supported by the National Institute of Infectious Diseases; and National Institute of Health Research Projects (#2022-ER1903-01)

Simultaneous Detection of EGFR and VEGF in Colorectal Cancer using Fluorescence-Raman Endoscopy

Fluorescence endomicroscopy provides quick access to molecular targets, while Raman spectroscopy allows the detection of multiple molecular targets. Using a simultaneous fluorescence-Raman endoscopic system (FRES), we herein demonstrate its potential in cancer diagnosis in an orthotopically induced colorectal cancer (CRC) xenograft model. In the model, epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF) were targeted with antibody-conjugated fluorescence and surface-enhanced Raman scattering (F-SERS) dots. FRES demonstrated fast signal detection and multiplex targeting ability using fluorescence and Raman signals to detect the F-SERS dots. In addition, FRES showed a multiplex targeting ability even on a subcentimeter-sized CRC after spraying with a dose of 50 µg F-SERS dots. In conclusion, molecular characteristics of tumor cells (EGFR in cancer cell membranes) and tumor microenvironments (VEGF in the extracellular matrix) could be simultaneously investigated when performing a colonoscopy