Biallelic structural variants in three patients with ERCC8-related Cockayne syndrome and a potential pitfall of copy number variation analysis.

Cockayne syndrome (CS) is a rare autosomal recessive disorder caused by mutations in ERCC8 or ERCC6. Most pathogenic variants in ERCC8 are single nucleotide substitutions. Structural variants (SVs) have been reported in patients with ERCC8-related CS. However, comprehensive molecular detection, including SVs of ERCC8, in CS patients remains problematic. Herein, we present three Japanese patients with ERCC8-related CS in whom causative SVs were identified using whole-exome-based copy number variation (CNV) detection tools. One patient showed compound heterozygosity for a 259-kb deletion and a deletion of exon 4 which has previously been reported as an Asia-specific variant. The other two patients were homozygous for the same exon 4 deletion. The exon 4 deletion was detected only by the ExomeDepth software. Intrigued by the discrepancy in the detection capability of various tools for the SVs, we evaluated the analytic performance of four whole-exome-based CNV detection tools using an exome data set from 337 healthy individuals. A total of 1,278,141 exons were predicted as being affected by the 4 CNV tools. Interestingly 95.1% of these affected exons were detected by one tool alone. Thus, we expect that the use of multiple tools may improve the detection rate of SVs from aligned exome data.

Bactericidal Efficacy of the Combination of Maresin-like Proresolving Mediators and Carbenicillin Action on Biofilm-Forming Burn Trauma Infection-Related Bacteria.

Biofilm-associated bacterial infections are the major reason for treatment failure in many diseases including burn trauma infections. Uncontrolled inflammation induced by bacteria leads to materiality, tissue damage, and chronic diseases. Specialized proresolving mediators (SPMs), including maresin-like lipid mediators (MarLs), are enzymatically biosynthesized from omega-3 essential long-chain polyunsaturated fatty acids, especially docosahexaenoic acid (DHA), by macrophages and other leukocytes. SPMs exhibit strong inflammation-resolving activities, especially inflammation provoked by bacterial infection. In this study, we explored the potential direct inhibitory activities of three MarLs on Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa and Escherichia coli) bacteria in their biofilms that are leading bacteria in burn trauma-related infections. We also examined the effects of MarLs on the bactericidal activities of a typical broad-spectrum antibiotic, carbenicillin (carb), on these bacteria in their preformed biofilms. The results revealed that MarLs combined with carbenicillin can inhibit the survival of Gram-positive and Gram-negative bacteria in their biofilms although MarLs alone did not exhibit bactericidal activity. Thus, our findings suggest that the combination of MarLs and carbenicillin can lower the antibiotic requirements to kill the bacteria in preformed biofilms.

Synthesis of fulgidic acid and the two possible stereoisomers of chaenomic acid D.

We synthesized fulgidic acid and the proposed structure for chaenomic acid D. The core part of the two natural products was constructed stereoselectively by the addition of acetic acid to the α,ß-unsaturated epoxy alcohol in the presence of a palladium catalyst. Subsequently, the two natural products were synthesized from the intermediate in a few steps. The data for the synthesized fulgidic acid were in good agreement with the reported data. Chaenomic acid was in good agreement with the natural product in the 1H and 13C NMR data, but not in the optical rotation. The 15R-isomer of chaenomic acid was also synthesized, but the 1H and 13C NMR data did not agree with the natural product.

Identification and characterization of bioactive metabolites of 12-hydroxyheptadecatrienoic acid, a ligand for leukotriene B4 receptor 2.

12(S)-hydroxyheptadecatrienoic acid (12-HHT) is a bioactive fatty acid synthesized from arachidonic acid via the cyclooxygenase pathway and serves as an endogenous ligand for the low-affinity leukotriene B4 receptor 2 (BLT2). Although the 12-HHT/BLT2 axis contributes to the maintenance of epithelial homeostasis, 12-HHT metabolism under physiological conditions is unclear. In this study, 12-keto-heptadecatrienoic acid (12-KHT) and 10,11-dihydro-12-KHT (10,11dh-12-KHT) were detected as 12-HHT metabolites in the human megakaryocytic cell line MEG01s. We found that 12-KHT and 10,11dh-12-KHT are produced from 12-HHT by 15-hydroxyprostaglandin dehydrogenase (15-PGDH) and prostaglandin reductase 1 (PTGR1), key enzymes in the degradation of prostaglandins, respectively. The 15-PGDH inhibitor SW033291 completely suppressed the production of 12-KHT and 10,11dh-12-KHT in MEG01s cells, resulting in a 9-fold accumulation of 12-HHT. 12-KHT and 10,11dh-12-KHT were produced in mouse skin wounds, and the levels were significantly suppressed by SW033291. Surprisingly, the agonistic activities of 12-KHT and 10,11dh-12-KHT on BLT2 were comparable to that of 12-HHT. Taken together, 12-HHT is metabolized into 12-KHT by 15-PGDH, and then 10,11dh-12-KHT by PTGR1 without losing the agonistic activity.

Total synthesis of resolvin D3.

Resolvin D3 was synthesized by the Suzuki-Miyaura cross-coupling reaction of C1-C8 borane with C9-C22 iodoolefin as the key reaction. The latter intermediate was obtained by the sequential Wittig reactions of C9-C13 phosphonium salt with C14-C19 aldehyde and then C9-C19 aldehyde with propyltriphenylphosphonium bromide. The stereogenic centers at C4, C11, and C17 were constructed by the ruthenium-catalyzed asymmetric transfer hydrogenation with high stereoselectivity.

Isoflurane attenuates sepsis-associated lung injury.

Acute lung injury is one of major complications associated with sepsis, responsible for morbidity and mortality. Patients who suffer from acute lung injury often require respiratory support under sedations, and it would be important to know the role of sedatives in lung injury. We examined volatile anesthetic isoflurane, which is commonly used in surgical setting, but also used as an alternative sedative in intensive care settings in European countries and Canada. We found that isoflurane exposure attenuated neutrophil recruitment to the lungs in mice suffering from experimental polymicrobial abdominal sepsis. We found that isoflurane attenuated one of major neutrophil chemoattractants LTB4 mediated response via its receptor BLT1 in neutrophils. Furthermore, we have shown that isoflurane directly bound to BLT1 by a competition assay using newly developed labeled BLT1 antagonist, suggesting that isoflurane would be a BLT1 antagonist.

Association between perceived organizational support and COVID-19 vaccination intention: A cross-sectional study.

OBJECTIVES: We examined the association between perceived organizational support (POS) and COVID-19 vaccination intention as well as the influence of the implementation of workplace infection prevention measures. METHODS: We analyzed data from an Internet survey of workers aged 20-65 years conducted in December 2020, during a period of widespread COVID-19 infection in Japan. RESULTS: Of the 23 846 participants in this survey, 1958 (8%) reported very high POS. In the group with very high POS, 836 (43%) workers wanted the COVID-19 vaccination; in contrast 1382 (36%) workers in the group with very low POS wanted the vaccination. POS was associated with COVID-19 vaccination intention (odds ratio [OR] = 1.11). The OR decreased after additional adjustment for the number of workplace infection control measures (OR = 1.06). In the analysis with POS as a categorical variable, very high POS was associated with COVID-19 vaccination intention (reference: very low POS) (OR = 1.34). The OR decreased after additional adjustment for the number of workplace infection control measures (OR = 1.17). High POS was associated with COVID-19 vaccination intention (OR = 1.17) but not with vaccination intention after additional adjustment for the number of workplace infection control measures (OR = 1.05). CONCLUSIONS: High POS during the COVID-19 pandemic increased employees' vaccination intention. The relationship between POS and vaccination intention was strongly influenced by implementation of workplace infection prevention measures. Implementing appropriate workplace infection prevention measures in the event of an emerging infectious disease outbreak may influence the vaccination behavior of employees, which may contribute to the acquisition of herd immunity in the community.

Synthesis of a TNF inhibitor, flurbiprofen and an i-Pr analogue in enantioenriched forms by copper-catalyzed propargylic substitution with Grignard reagents.

The copper-catalyzed substitution reaction of diethyl phosphate derived from TMSCCCH(OH)CH2CH2OTBDPS with 3-c-C5H9-4-MeOC6H3MgBr, followed by several transformations, afforded a tumor necrosis factor inhibitor possessing a Ph-acetylene moiety. The inhibitor was also synthesized from phenylacetylene phosphate PhCCCH(OP(O)(OEt)2)CH2CH2OTBDPS. Furthermore, the substitution of phosphates derived from TMSCCCH(OH)CH3 and TMSCCCH(OH)-i-Pr with 3-F-4-PhC6H3MgBr gave the corresponding substitution products, which were transformed to flurbiprofen and its i-Pr analogue, respectively. The copper-catalyzed substitutions in these syntheses proceeded in a regio- and stereoselective manner.

[Qualitative analysis of provided information and advice from occupational physicians to attending clinical physicians in supporting an employee's work-treatment balance].

OBJECTIVES: In Japan, the population is aging and there is a declining birth rate. It is an important occupational health issue to support the balance between illness treatment (including nursing care, childcare, etc.) and work. Many patients require mental and financial support to help them with their work-treatment balance. In 2016, the Ministry of Health, Labor and Welfare provided guidelines for supporting employee's work-treatment balance, and in 2018, "Consulting Fee" was approved as an insured medical treatment when clinic doctors supported their patients for continuing to work. The request for the consulting fee requires that the clinician and the occupational physician exchange information on the support necessary for the patient to continue working. Generally, occupational physicians obtain medical information from clinicians to give advice on a worker's employment considerations. However, we do not know what kind of workplace information clinicians hope to know when treating their patients. Therefore, we conducted this survey to clarify how occupational physicians could provide useful information to clinicians. METHODS: We asked approximately 1,500 occupational physicians from the Occupational Health Subcommittee of the Japan Society for Occupational Health to provide us with a letter sent to their clinician to assist workers. From the collected letters, the structural parts of the letters (titles, greetings, acknowledgments, etc.) were removed. We defined a section as a contextual unit that does not impair the meaning. The prepared sections underwent qualitative inductive analysis using the content analysis method of "Berelson, B." RESULTS: A total of 103 cases and 178 documents from 42 people were included in the analysis. Extracting descriptions that could be interpreted as providing information, including descriptions related to treatment, employment, and living environment, and opinions and suggestions from occupational physicians resulted in 596 sections. As a result of the qualitative and inductive classification, the information was classified into three large categories that consisted of information provision, opinions of occupational physicians, and information handling, five middle and eighteen small classifications. In addition, some good practices that were considered significant to clinicians were illustrated. CONCLUSIONS: We analyzed and categorized the information present in the letters sent by occupational physicians to clinicians. The letter does not need to contain all the information in the category table. However, it is important that it should have the necessary and sufficient information considering the case in question. We believe that this category table will aid occupational physicians in writing letters to clinicians.

α- and γ-Regiocontrol and Enantiospecificity in the Copper-Catalyzed Substitution Reaction of Propargylic Phosphates with Grignard Reagents.

The regioselectivity (r.s.) and enantiospecificity (e.s.) of the substitution reactions of secondary propargylic alcohol derivatives using reagents derived from ArMgBr and Cu salts were studied. First, the picolinate, 3-methylpicolinate, and diethylphosphonate derivatives of Ph(CH2 )2 CH(OH)C≡CTMS were reacted with PhMgBr/CuCN in ratios of 2.5:2.7-2.5:0.25. The use of 2.5:0.25 ratio in THF/DME (6:1) at 0 °C for 1 h afforded the α-substitution product from the phosphate with ≥98 % r.s. and 99 % e.s. CuBr⋅Me2 S gave similar selectivity. The reaction system was then applied to phosphates derived from R1 CH(OH)C≡CR2 and ArMgBr to obtain synthetically sufficient r.s. and e.s. values with R2 =TMS, Ph, whereas iPr was borderline in terms of size as an R1 substituent. The presence of a substituent at the o-position of Ar marginally affected the selectivity. We also found that the use of PhMgBr/Cu(acac)2 in a 2:1 ratio in THF produced the γ-substitution products (allenes) with high r.s. and e.s.

Follow-Up of Occupational Health Issues and Measures Taken in Fukushima Daiichi Nuclear Power Plant Where Decommissioning Work Has Continued Over 6 Years Since 2014.

OBJECTIVES: To follow up arising occupational health (OH) issues, measures taken, and their performances in the Fukushima Daiichi Nuclear Power Plant since 2014, and thus share experiences and extend the contribution of OH to long-term decommissioning work and preparation for future disasters. METHODS: Necessary information from official reports and through the OH-related activities involved was collected and analyzed. RESULTS: The issues were categorized into establishment of the OH management system, three individual issues, and others. During the 6 years until end 2019, the OH management system has been strengthened and OH measures have been enriched gradually by visualizing the broader picture, even though some resistance and problems have been encountered. CONCLUSIONS: Improvement in the autonomy of contractors and their ability to respond to environmental changes is necessary.

Assessing the effect of mandatory progress reporting on treatment requirements identified during health examinations at the Fukushima Daiichi Nuclear Power Plant: A time series analysis.

OBJECTIVES: At the Fukushima Daiichi Nuclear Power Plant, run by the Tokyo Electric Power Company, new procedures were introduced as part of the fitness for duty program in July 2016. These were designed to ensure that treatment and further investigations identified as necessary during health examinations were carried out. This study aimed to assess the effectiveness of the initiative by testing whether workers who needed further health examinations obtained them promptly, and whether the number with unmet health needs decreased and the number of workers being treated increased. METHODS: The primary contractors reported aggregated quarterly results of health examinations of both their own and their subcontractors' employees, and follow-up visits to medical institutions were also reported over the next two quarters. The study used data for the period from July 2016 to December 2018. Incident rate ratios were estimated using a multilevel Poisson regression model, including the logarithm of the number of workers who took health examination for each primary contractor company as offset. The linear trend was assessed by treating the number of periods as a continuous variable. RESULTS: The incident rate ratio for workers who needed treatment having a follow-up examination promptly showed a significant decrease over time. The incident rate ratio for those with unmet needs decreased, and those being treated increased over time. CONCLUSIONS: The findings showed that the initiative was effective, with the number of early visits for further health examinations increasing and a decrease in the number of people with unmet health needs.

Synthesis of Two Stereoisomers of Potentially Bioactive 13,19,20-Trihydroxy Derivative of Docosahexaenoic Acid.

The C16-C22 fragment with the acetylene terminus was constructed through the asymmetric dihydroxylation of the corresponding olefin, while the 15-iodo-olefin corresponding to the C11-C15 part was prepared via the asymmetric transfer hydrogenation of the corresponding acetylene ketone followed by hydrozirconation/iodination. Both pieces were joined by a Sonogashira coupling, and the product was further converted into the title compound via a Wittig reaction with the remaining C1-C10 segment and Boland reduction using Zn with TMSCl.

[Systems and human resource development for occupational health in India: Effective occupational health management for Japanese enterprises with overseas branches].

OBJECTIVES: This study aimed to collect and assess information about occupational health in India, for Japanese enterprises. METHODS: We conducted a preliminary survey through literature reviews and internet search engines. We then conducted interview-based surveys at a central government agency, an international organization, the Japanese embassy, educational and research institutions, and Japanese enterprises in India. This information was categorized into: (1) organizations, legal and inspection systems in occupational health, (2) occupational health management and specialists in workplaces, (3) occupational health-related activities in workplaces resulting from legal obligations, and (4) healthcare and workers' compensation systems. RESULTS: The Indian Ministry of Labour and Employment is primarily responsible for occupational safety and health. There are four main acts of legislation covering occupational safety and health in the factories, ports, mines, and construction sites. The Factories Act, 1948, mandates the establishment of an occupational health center and a safety committee in the factories; the appointment of factory medical officers and safety officers. These medical officers must possess an Indian medical degree, and undertake a three months' course to obtain an Associate Fellow of Industrial Health certificate. The rules and regulations under this act differ in each Indian state. Low-wage workers are registered with a medical insurance scheme. Most workers are covered by workers' compensation schemes, although the number of reported occupational injuries are low. CONCLUSIONS: Japanese enterprises should consider the local conditions of occupational health in India because of the different legalities and occupational health status in each state. Regardless of the Factories Act, 1948, stipulating a variety of occupational health-related activities, inadequate legal compliance is suspected to be common because of the ineffective labor inspection requirements and a shortage of specialists on human resources. The study also revealed a deficient social security system. Therefore, the internal educational support for specialists, external support from the company headquarters in Japan, and the local institutions in India; and the systemic support for effective occupational health-related activities are required for improving the status of occupational health in the factories in India.

[System and human resources for occupational health in the republic of the Philippines: Management of occupational health activities at overseas workplaces for Japanese enterprises].

OBJECTIVES: This study aimed to consider the appropriate occupational health system for Japanese enterprises in the Philippines based on information on the regulations and development of specialists. METHODS: We collected information using an information-gathering checklist. Along with literature and internet surveys, we conducted interviews by visiting local business sites, central government agencies in charge of medical and health issues, and educational institutions with specialized occupational physician training curricula. RESULTS: Occupational health administration in the Philippines is managed by the Department of Labor and Employment, which issues the Occupational Safety and Health Standards that specify the legal requirements for occupational health. A new law(Republic Act 11058),enacted in 2018 to strengthen the Occupational Safety and Health Standards, has newly established a penalty provision in case of violations. Professional personnel responsible for occupational health are grouped as safety officers and occupational health personnel, including occupational physicians and occupational nurses; training is conducted at the Occupational Safety and Health Center of the Department of Labor and Employment and educational institutions. The basic medical insurance system and the workers' compensation system are operated by the Philippine Health Insurance Corporation and Social Security Committee, respectively, both of which are government agencies. CONCLUSIONS: We confirmed that occupational health activities in the Philippines are based on government regulations, namely, the Occupational Safety and Health Standards. In addition, the enactment of a new law calls for strict compliance with corporate occupational health activities. To manage proper occupational health activities at overseas workplaces, Japanese corporations should clarify corporate-wide policies and support local employers in complying with regulations and utilizing highly specialized personnel.

[Systems and actual practices of occupational safety and health risk management in the People's Republic of China].

OBJECTIVES: To investigate the systems and actual practices of occupational safety and health risk management in China. METHODS: First, we gathered basic information through a literature review using an academic search engine (Japan Medical Abstracts society, Pubmed, and Google Scholar), as well as a general search on the Internet. Next, we conducted field surveys at a graduate school for public health, providers of occupational health services (e.g. medical examinations, working environment measurements), and local workplaces of a Japanese construction machinery company in China. This information was analyzed in terms of legal framework, professional staff, working environment measurements, medical examinations, occupational diseases, and occupational health service providers. RESULTS: Health and safety-related matters have become codified in Chinese workplaces as a result of safety laws and measures to prevent occupational diseases. While the country does have safety and hygiene officers, they lack official frameworks for occupational physicians and nursing professionals. The employers are not obligated to appoint medical professionals. While general medical examinations are not provided for under Chinese law, businesses are obligated to bring in external providers of occupational safety to perform special medical exams and working environment measurements. Occupational diseases are on the rise; pneumoconiosis comprises roughly 80% of cases. In addition, occupational health technical service providers have specialized staff and are not permitted to perform medical examinations or other services without government accreditation. DISCUSSION/CONCLUSION: There are great disparities in specialist knowledge about health and hygiene between company staff and external organizations, thus running the risk of corporate health and safety policies existing only on paper. This issue demands greater utilization of public health physicians in Chinese workplaces and support from Japanese professionals who understand how occupational safety and health risk management operate in China.

Developing a global occupational health and safety management system model for Japanese companies.

OBJECTIVES: To develop and validate a global occupational health and safety management system (OHSMS) model for Japanese companies. METHODS: In cooperation with a Japanese company, we established a research team and gathered information on occupational health and safety (OHS) practices in nine countries where the target company operated manufacturing sites. We then developed a model hypothesis via research team meeting. The model hypothesis was introduced to local factories in Indonesia and Thailand as trial sites. We evaluated the roles of the company headquarters, the implementation process, and any improvements in OHS practices at the sites. Based on the results, a global OHSMS model was formalized for global introduction. RESULTS: The model consisted of both headquarters and site roles. These roles were well-functioning, and OHS at the sites improved. Two issues concerning the functioning of the headquarters were identified: the need to establish a reporting system to the headquarters and the need to support the improvement of specialized human resources. By improving the model hypothesis to address these issues, the model was formalized for global introduction. CONCLUSIONS: The global OHSMS model was based on the use of methods and specialized human resources relevant to each region and their common objectives, as well as evaluation indicators based on the minimum requirements of the company headquarters. To verify the effectiveness of this model, the experiment should be extended to other countries.

Stereoselective Synthesis of Maresin-like Lipid Mediators.

Maresin-L1 (14S,22-dihydroxy-docosa-4Z,7Z,10Z,12E,16Z,19Z-hexaenoic acid) and maresin-L2 (14R,22-dihydroxy-docosa-4Z,7Z,10Z,12E,16Z,19Z-hexaenoic acid) were chemically synthesized. They were identical to activated macrophage produced counterparts and their total synthesis was highly Stereoselective, as revealed by chiral LC-UV-MS/MS analysis. The synthesis involved the following steps: (1) kinetic resolution of a racemic allylic alcohol by the asymmetric epoxidation; (2) transformation of the epoxy alcohol to γ-hydroxyenal derivative; and (3) the Wittig reaction to furnish the Z-olefin.

Use of the 2-Pyridinesulfonyloxy Leaving Group for the Fast Copper-Catalyzed Coupling Reaction at Secondary Alkyl Carbons with Grignard Reagents.

Investigation of the copper-catalyzed coupling reaction of 2-pyridinesulfonates with Grignard reagents revealed that reactions with catalytic Cu(OTf)2 were completed in <40 min. The results differed from those of the previous CuI-catalyzed reactions of tosylates in the presence of additives (LiOMe and TMEDA) for 12-24 h. It was shown that the preferred coordination of the leaving group to the reagents accelerated the reaction. Successful reagents were MeMgCl and other RMgX. Complete inversion was established.

Synthesis of resolvin E3 via palladium-catalyzed addition of AcOH to vinyl epoxy alcohols.

(18R)- and (18S)-stereoisomers of resolvin E3 (RvE3), potent anti-inflammatory mediators, were synthesized stereo- and enantioselectively through the Wittig reaction of the carbonate of 6R,7R- and 6R,7S-dihydroxynona-2E,4E-dienal, a C12-C20 part, with the phosphonium salt corresponding to the C1-C11 part. The stereoisomeric carbonate was prepared by the Swern oxidation of 3-(AcO)-6R,7R- or 3-(AcO)-6R,7S-(dihydroxy-carbonate)-4-nonen-1-ol followed by the spontaneous elimination of the AcO group in one pot. The (6R,7R)-(dihydroxy-carbonate)-alcohol for (18R)-RvE3 was, in turn, provided by stereoselective epoxidation of 9-(TBS-oxy)nona-4Z,6E-dien-3R-ol with m-CPBA and the subsequent Pd-catalyzed addition of AcOH to the resulting syn vinyl epoxy alcohol followed by carbonate formation of the vic-syn-diol and TBS desilylation. The Mitsunobu inversion of the syn vinyl epoxy alcohol gave the anti isomer, which was converted to 3-(AcO)-6R,7S-(dihydroxy-carbonate)-4-nonen-1-ol, the intermediate to (18S)-RvE3, by the same set of reactions.