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Shiga toxin-producing Escherichia coli (STEC) are associated with severe infections including hemorrhagic colitis and hemolytic uremic syndrome in humans. Ruminants are known as reservoirs of STEC; however, no data are available on STEC in ruminants in Mongolia, where more than 5 million cattle and 25 million sheep are raised. To disclose the existence and characteristics of STEC in Mongolia, in this study, we isolated and characterized STEC from cattle in Mongolia. We collected 350 rectal swabs of cattle from 30 farms near Ulaanbaatar city and isolated 45 STEC from 21 farms. Rectal swabs were precultured with modified Escherichia coli broth and then inoculated to Cefixime-Tellurite Sorbitol MacConkey agar plate and/or CHROMagar STEC agar plate for the isolation of STEC. The isolation ratios in each farm were from 0% to 40%. Multiplex PCR for the estimation of O- and H-serotypes identified 12 O-genotypes (Og-types) and 11 H-genotypes (Hg-types) from 45 isolates; however, Og-types of 19 isolates could not be determined. Stx gene subtyping by PCR identified 2 stx1 subtypes (1a and 1c) and 4 stx2 subtypes (2a, 2c, 2d, and 2g). Forty-five isolates were divided into 21 different groups based on the Og- and Hg-types, stx gene subtypes and the existence of virulence factors, ehxA, eae, and saa, which includes several major serotypes associated with human illness such as O26:H11 and O157:H7. The most dominant isolate, OgUT:H19 [stx1a (+), stx2a (+), ehxA (+) and saa (+)], was isolated from eight farms. This is the first report on the characterization of STEC in cattle in Mongolia, and the results suggest the importance of further monitoring of STEC contamination in the food chains as well as STEC infection in humans.

Abstract Background Abnormal behavior after oseltamivir administration has been reported in the media; in 2007, the package insert for oseltamivir phosphate was revised to restrict its administration to individuals aged over 10 years. However, in 2018, the age limitation specified in the package insert was removed. Here, we evaluated the trends in anti-influenza drug prescription and adverse drug reactions (ADRs) reported in pediatric outpatients after revising the oseltamivir package insert as an ecological study. Methods Anti-influenza drug prescriptions for pediatric outpatients with influenza aged 0–19 years were downloaded from the acute Diagnosis Procedure Combination hospital databases using the MDV analyzer®. ADR reports on anti-influenza drug prescription among patients aged 0–20 years in the Japanese Adverse Drug Event Report database were downloaded from the Pharmaceutical and Medical Devices Agency website. Data were collected during the 2016/2017 and 2019/2020 influenza seasons. Results During the influenza epidemic season (January–March), the percentage of oseltamivir prescriptions for patients with influenza aged 10–19 years tripled after the revision of the oseltamivir package insert (9.3% during the 2016/2017 season and 29.2% during the 2019/2020 season); however, reports of abnormal behavior did not increase (two during the 2016/2017 season and none during the 2019/2020 season). Conclusions The number of oseltamivir-related ADR reports among minors over 10 years of age did not increase although the proportion of oseltamivir prescriptions increased after the revision of the oseltamivir package insert.

Abstract Background Esophagectomy is a high-risk procedure that can involve serious postoperative complications. There has been an increase in the number of minimally invasive esophagectomies (MIEs) being performed. However, the relationship between intraoperative management and postoperative complications in MIE remains unclear. Methods After the institutional review board approval, we enrolled 300 patients who underwent MIE at Tohoku University Hospital between April 2016 and March 2021. The relationships among patient characteristics, intraoperative and perioperative factors, and postoperative complications were retrospectively analyzed. The primary outcome was the relationship between intraoperative fluid volume and anastomotic leakage, and the secondary outcomes included the associations between other perioperative factors and postoperative complications. Results Among 300 patients, 28 were excluded because of missing data; accordingly, 272 patients were included in the final analysis. The median [interquartile range] operative duration was 599 [545–682] minutes; total intraoperative infusion volume was 3,747 [3,038–4,399] mL; total infusion volume per body weight per hour was 5.48 [4.42–6.73] mL/kg/h; and fluid balance was + 2,648 [2,015–3,263] mL. The postoperative complications included anastomotic leakage in 68 (25%) patients, recurrent nerve palsy in 91 (33%) patients, pneumonia in 62 (23%) patients, cardiac arrhythmia in 13 (5%) patients, acute kidney injury in 5 (2%) patients, and heart failure in 5 (2%) patients. The Cochrane-Armitage trend test indicated significantly increased anastomotic leakage among patients with a relatively high total infusion volume (P = 0.0085). Moreover, anastomotic leakage was associated with male sex but not with peak serum lactate levels. Patients with a longer anesthesia duration or recurrent nerve palsy had a significantly higher incidence of postoperative pneumonia than those without. Further, the incidence of postoperative pneumonia was not associated with the operative duration, total infusion volume, or fluid balance. The operative duration and blood loss were related to the total infusion volume. Acute kidney injury was not associated with the total infusion volume or serum lactate levels. Conclusions Among patients who underwent MIE, the total infusion volume was positively correlated with the incidence of anastomotic leakage. Further, postoperative pneumonia was associated with recurrent nerve palsy but not total infusion volume or fluid balance.

Background: Functional deterioration following emergency general surgery (EGS) poses a significant challenge in super-elderly patients. However, limited research has focused on assessing the deterioration in postoperative performance status (PS). This study aimed to investigate the impact of EGS on PS deterioration in super-elderly patients, and the extent to which deteriorated PS is recovered. Methods: This historical cohort study comprised 77 super-elderly patients who underwent EGS between July 2015 and December 2020. Functional deterioration was evaluated by comparing preoperative and postoperative Eastern Cooperative Oncology Group Performance Status (ECOG-PS). The Emergency Surgical Score (ESS) was used as a risk-adjustment tool. Questionnaires were mailed to the patients and their families to assess post-discharge PS and obtain their impressions of EGS. Results: Postoperative PS deteriorated in 35/77 patients (45.5 %). Significant differences were observed between the groups in terms of sex, serum C-reactive protein (CRP) levels, ESS scores, preoperative ECOG-PS, duration of operation, and major complications. Multivariate analysis of preoperative factors showed that ESS ≥7 (OR: 3.7, 95 % CI: 1.0–13), preoperative ECOG-PS ≤2 (OR: 5.9, 95 % CI: 1.7–21), and female sex (OR: 5.8, 95 % CI: 1.6–21) were associated with postoperative ECOG-PS deterioration. According to the questionnaire results, PS recovery post-discharge was observed in 6/36 (17 %) patients, and 34/36 (94 %) patients and their families expressed positive impressions of EGS. Conclusions: EGS in super-elderly patients highly caused a deterioration in their PS, particularly in patients with maintained preoperative PS. PS hardly recovered; however, most patients and their families had positive impressions of the EGS. Key message: We assessed the pre- and postoperative performance status of super-elderly patients who underwent emergency general surgery. Surgery caused a marked deterioration in patients' functional performance, which seldom recovered postoperatively.

Three-dimensional eddy current distributions in cold crucibles are analyzed by using an integro-differential method. The electromagnetic force is calculated from the obtained eddy current distributions. In this paper, computations of eddy current and levitation force of the molten metal in some cold crucible models are performed, and the influences of coil position, the number of segments of the crucible, frequency of the coil current and gap length between the segments on the levitation force are discussed. Furthermore, an experimenta verification is presented.

ABSTRACT Aim/Introduction Resistin, which induces insulin resistance, is mainly expressed in monocytes/macrophages in humans. We reported previously that serum resistin was highest in the G–A haplotype defined by resistin single nucleotide polymorphisms (SNPs) at −420 (rs1862513) and − 358 (rs3219175). As sarcopenic obesity is associated with insulin resistance, we aimed to examine whether serum resistin and its haplotypes were associated with sarcopenic obesity at a latent stage. Materials and Methods We cross‐sectionally analyzed 567 community‐dwelling Japanese participants attending annual medical check‐ups in which the sarcopenic obesity index was evaluated. The age‐ and gender‐matched normal glucose tolerance subjects with G–A homozygotes and those with C–G homozygotes were examined via RNA‐sequencing and pathway analysis (each n = 3), and RT‐PCR (each n = 8). Results In multivariate logistic regression analyses, the fourth quartile (Q4) of serum resistin and G–A homozygotes were both associated with the latent sarcopenic obesity index defined by a visceral fat area of ≥ 100 cm2 and grip strength Q1 after adjustment for age and gender, with or without other confounding factors. RNA sequencing and pathway analysis showed that tumor necrosis factor (TNF) was involved in the top five pathways in the whole blood cells of G–A homozygotes compared with C–G homozygotes. RT‐PCR revealed that TNF mRNA was higher in G–A homozygotes than in C–G homozygotes. Conclusions The G–A haplotype was associated with the latent sarcopenic obesity index defined by grip strength in the Japanese cohort, could be mediated by TNF‐α.

In the sinusoidal steady-state analysis, it is an advantage of boundary element method that eddy current analysis can be performed by only the surface integral without the volume integral. However, the volume integral and the surface integral are necessary for boundary element analysis in the transient problem. Therefore, the advantage of the boundary element method vanishes. In this paper, techniques of the transient analysis by boundary element method using the Fourier transform are presented. In the proposed method, the response of the frequency domain is obtained by boundary element analysis in the sinusoidal steady-state analysis. Transient analysis is by the inverse Fourier transform using the response of the frequency domain. Therefore, the volume integral with respect to the region to be analyzed is not necessary. The applicability of the presented method is shown by the comparison of the computation results of the two-dimensional model and three-dimensional model with the theoretical values. Furthermore, the stability of the presented method is compared with the time differential method.

In order to reduce computer resources for the analysis of eddy current distribution of cold crucible models, the authors employed a current sheet approximation for eddy current distribution because penetration depth of the eddy current is relatively small in comparison with the dimension of the conducting bodies. The eddy current distributions approximated by current sheets are solved for by an integro-differential method using an electric vector potential. Furthermore, magnetic flux density, power loss, Lorentz force and lifting force of the molten metals are obtained from the solved eddy current distribution.

Objectives: In stroke patients, the assessment of gait ability over time is important. For quantitative gait assessment using measuring devices, the walking speed condition for measurement is generally based on the patient’s preferred walking speed or the maximum walking speed at the time of measurement. However, because walking speed often increases during the convalescent stage, understanding the effects of change in walking speed on gait when comparing the course of recovery is necessary. Although several previous studies have reported the effects of change in walking speed on gait in stroke patients, the time-distance parameters described in these reports may not be generalizable because of the small case numbers. Therefore, we measured treadmill gait at the preferred walking speed (PWS) and 1.3 times the PWS (130% PWS) in 43 post-stroke hemiplegic patients and analyzed the effects of change in walking speed on time-distance parameters. Methods: Forty-three patients with hemiplegia after a first stroke, who were able to walk on a treadmill under supervision, were recruited as subjects. Using a three-dimensional motion analysis system, treadmill gait was assessed under two conditions: PWS and 130% PWS. The primary outcome measures were the time-distance parameters, which were compared between the PWS and 130% PWS conditions. Results: Cadence, stride length, and step length of the affected and unaffected lower limbs increased significantly at 130% PWS compared with at PWS. In terms of actual time, single stance time and initial and terminal double stance time in both affected and unaffected limbs decreased significantly at 130% PWS. In terms of relative time (% of the gait cycle), compared with PWS, relative single stance time increased significantly, whereas relative initial and terminal double stance times decreased significantly at 130% PWS in both the affected and unaffected limbs. Conclusions: This study on treadmill gait in patients with hemiplegia after a first stroke confirmed the effects of change in walking speed on time-distance parameters. Our results will help in the interpretation of time-distance parameters measured under different walking speed conditions.

Abstract Although FOLFIRINOX (l-Leucovorin/5-FU/Irinotecan/Oxaliplatin) is established as one of the standard therapies for patients with metastatic pancreatic cancer, the modified FOLFIRINOX (mFOLFIRINOX) is often used in clinical practice to reduce the incidence of toxicities. Febrile neutropenia (FN) and severe neutropenia during FOLFIRINOX are especially frequently observed in Japanese patients. In this study, we evaluated the incidence of FN and severe neutropenia, and explored the risk factors for severe neutropenia in patients receiving treatment with mFOLFIRINOX. The data of patients who had received mFOLFIRINOX between December 2013 and December 2014 at the National Cancer Center Hospital East were reviewed retrospectively. We graded the neutropenia severity and defined ≥ Grade 3 neutropenia as severe neutropenia. Univariate and multivariate analysis were undertaken to evaluate the associations with risk of development of severe neutropenia. A total of 122 patients were enrolled in this study. Sixty two patients (51%) and 10 patients (8%) developed severe neutropenia and FN, respectively. Multivariate analysis identified a low baseline white blood cell count (odds ratio [OR], 14.50; 95% confidence interval (CI), 3.27–111.14; p = 0.002) and presence of heterozygosity for UGT1A1*28 or UGT1A1*6 polymorphism (OR, 2.84; 95% CI, 1.18–7.17; p = 0.023) as independent risk factors for severe neutropenia. The incidences of severe neutropenia and FN in patients receiving mFOLFIRINOX in our clinical practice were comparable to previous reports. The risk factors for severe neutropenia in patients receiving mFOLFIRINOX were a low baseline white blood cell count and presence of heterozygosity for UGT1A1*28 or UGT1A1*6 polymorphism.

Cleavage of the glycan-branching enzyme N-acetylglucosaminyltransferase-V (GnT-V) by signal peptide peptidase-like 3 (SPPL3) protease and extracellular secretion of active glycan GnT-V depend on GnT-V’s own glycosylation state.

Byline: M. Egawa, M. Fukuda, H. Takashima, T. Misaki, K. Kinuya, S. Terahata A sentinel node (SN) is defined as the first site where cancer cells are carried by lymph [...]

Non-native species pose a threat to native organisms. When non-native and native species are closely related, the former can often competitively exclude the latter. Many studies have focused on competitive exclusion of native insect species by non-native eusocial hymenopterans, including ants, hornets, paper wasps and bees. Although solitary species of wasps have been introduced in many regions, few studies have investigated the effects of these insects on their native congeners. We investigated competitive interactions between native and non-native solitary wasps belonging to the same genus (Hymenoptera: Vespidae: Eumeninae: Anterhynchium). Specifically, we compared resource use and natural enemies of the native Anterhynchium flavomarginatum and supposedly non-native A. gibbifrons at a forest edge in Takasago, Hyogo, Japan, in June-October 2019, using trap nests (bamboo canes). Of 950 bamboo canes, 70 (7.4%) and 50 (5.3%) were used as nests by A. flavomarginatum and A. gibbifrons, respectively. Anterhynchium flavomarginatum produced two generations over the period studied, whereas A. gibbifrons produced only one. Although A. gibbifrons began nesting two weeks after A. flavomarginatum, the nesting period of A. gibbifrons overlapped that of the first nesting period of A. flavomarginatum. Nest architecture and the inner diameter of the canes used by both species were similar, suggesting potential competition for nesting resources. Anterhynchium flavomarginatum used larvae of 14 species of moths (Lepidoptera: Crambidae, Pyralidae, Tortricidae) as food for their larval offspring, whereas A. gibbifrons used only a single species, Demobotys pervulgalis (Lepidoptera: Crambidae). Prey species were exclusive to each wasp species, indicating no competition for this resource. Three parasitoid species, Macrosiagon nasutum (Coleoptera: Ripiphoridae), Amobia distorta (Diptera: Sarcophagidae) and Megaselia sp. (Diptera: Phoridae), attacked both Anterhynchium species. The percentage parasitism by Amobia distorta was higher for the native A. flavomarginatum. Anterhynchium gibbifrons may indirectly affect A. flavomarginatum via shared parasitoids.

Abstract Background Early gastric cancer (EGC) is often associated with lymphatic metastasis, but it is extremely rare to be found as a single giant lymph node. Cancer often becomes more malignant in metastatic lesions than in primary lesions, and retrodifferentiation to the fetal gastrointestinal tract during the metastatic process has been reported in gastric cancer. We report an extremely rare case of EGC with a 13-cm giant lymph node metastasis in which an adenocarcinoma with enteroblastic differentiation and yolk sac tumor-like components was observed. Case presentation The case was a 70-year-old man who visited his local doctor with right hypochondrial pain, which was identified by computed tomography (CT) as a giant mass. Upper endoscopy revealed a 30-mm-sized 0-IIc lesion in the greater curvature of the angular incisure and a 15-mm-sized 0-IIa lesion in the anterior wall of the lower body of the gastric body. Endoscopic biopsy revealed tubular adenocarcinoma in both lesions. The gastric lesion and the giant tumor were clinically regarded as independent lesions (gastrointestinal stromal tumor, [GIST], and EGCs), and distal gastrectomy and D1 + dissection were performed to comprehensively treat all lesions. Pathological examination revealed that the giant tumor was tubular adenocarcinoma with an intestinal phenotype and was considered a lymph node metastasis of EGCs. To exclude the possibility of metastasis of adenocarcinoma other than EGCs, postoperative positron emission tomography-computed tomography (PET-CT) and colonoscopy were performed; however, no primary site other than the stomach was found. Metastatic lymph nodes have an increased degree of atypia compared with the primary tumor, and yolk sac tumor-like carcinoma morphology was observed along with α-fetoprotein (AFP) and Spalt-like 4 (SALL4) expression in this case. It was considered that retrodifferentiation to a fetal phenotype occurred during the metastatic process. Liver metastasis occurred 6 months after surgery, and chemotherapy is currently being introduced. Conclusions We experienced a case of EGC with a single giant lymph node metastasis. Retrodifferentiation to the fetal gastrointestinal tract during metastasis was speculated to be involved in the formation of giant lymph node metastasis and liver metastasis in this case.

Background and Objectives Immune checkpoint inhibitors (ICIs) are effective in various types of cancer and cause immune-related adverse events (irAEs). The occurrence of irAEs is associated with improved survival outcome. We investigated the association between the occurrence of irAEs and overall survival (OS) and progression free survival (PFS), and the risk factors for the development of irAEs, in patients with non–small-cell lung cancer (NSCLC), gastric cancer (GC) and melanoma (MM) treated with ICIs. Methods This was a retrospective observational cohort study, and the data were taken from inpatients in a hospital. OS and PFS were compared among patients with different numbers of irAEs. Log-rank test and Cox regression and logistic regression analysis were applied, and details of irAEs characteristics were summarized. Results We obtained data from 200 patients. The major tumor types were NSCLC, GC, and MM. Median OS and PFS in all patients were 9.3 and 3.5 months, respectively. Patients without irAEs tended to have shorter OS or PFS compared with those with a single irAE or multi-system irAEs. Covariate analysis suggested that age (≥75 years), albumin (≥3.5 g/dL) and smoking history were significant for increased occurrence of irAEs. Pneumonitis and thyroiditis tended to occur frequently in patients with NSCLC and MM. The irAE grade was ≤2 in 67.3% of all irAEs, and days of irAEs onset varied. Conclusion We observed patients with irAEs tended to have better OS or PFS in patients with various types of cancers treated with ICIs. We suggest that ICIs should be used appropriately by continuously monitoring the irAEs.

Introduction: Current severity classification of appendicitis does not take into account the impact of bloodstream infection (BSI), despite the importance of BSI in critical care settings. Therefore, we examined whether BSI in emergency appendectomy indicated for acute appendicitis has a clinical impact. Methods: We retrospectively reviewed patients with acute appendicitis who underwent emergency surgery at our department between July 2015 and January 2020. Results: BSI was detected in 14 out of 154 patients (9%). Patients with BSI had a higher rate of major complications, readmission rate, and longer postoperative hospital stay, but there was no difference in the rate of complicated and uncomplicated appendicitis. Multivariate analysis showed age ≧ 58 years (Odds ratio [OR] 8.98 95% Confidence interval [CI] 2.07–39.0) and bilirubin ≧ 2.0 mg/dl (OR 8.79 95% CI 1.09–71.0) were associated with BSI. In 4 out of 8 cases (50%), the bacteria isolated in patients with BSI were not isolated in patients with intraabdominal infection (IAI). Conclusion: BSI in acute appendicitis had a clinical impact on postoperative outcomes, irrespective of complicated and uncomplicated appendicitis. Blood cultures should be considered for older patients, and patients with high bilirubin levels in emergency appendectomy for acute appendicitis.

Abstract We have developed a novel method to predict the success of PCR amplification for a specific primer set and DNA template based on the relationship between the primer sequence and the template. To perform the prediction using a recurrent neural network, the usual double-stranded formation between the primer and template nucleotide sequences was herein expressed as a five-lettered word. The set of words (pseudo-sentences) was placed to indicate the success or failure of PCR targeted to learn recurrent neural network (RNN). After learning pseudo-sentences, RNN predicted PCR results from pseudo-sentences which were created by primer and template sequences with 70% accuracy. These results suggest that PCR results could be predicted using learned RNN and the trained RNN could be used as a replacement for preliminary PCR experimentation. This is the first report which utilized the application of neural network for primer design and prediction of PCR results.

Controlling the alignment of blue phase liquid crystals is challenging. Here, electric-field induced phase transitions are exploited to control the crystal orientation of blue phase I, and applied to the fabrication of holograms with omnidirectional circular-polarization selectivity.

A dietary symbiotic supplement of okara, a by-product of tofu manufacturing with Bacillus coagulans lilac-01 found to reduce secondary bile acid (BA) in a rat study and cholic acid (CA), a primary 12α-hydroxylated (12αOH) BA, induces lipid accumulation in rats. In this study, we investigated whether dietary supplementation of a synbiotic with okara and Bacillus coagulans lilac-01 improves CA-induced hepatic lipid accumulation. Male Wistar/ST rats (4 weeks old) were fed with or without CA-supplementation combined with the synbiotic for 2 weeks. The CA diet increased hepatic triglycerides and the synbiotic diet in combination with CA normalized liver triglyceride concentration accompanied by suppression of hepatic fatty acid synthase expression, enhancement of fecal triglyceride excretion, and increase in fecal non-12αOH BA excretion. Dietary supplementation with the okara synbiotic ameliorated hepatic lipid accumulation probably by reducing de novo lipogenesis and enhancing fecal triglyceride excretion.

Fish and omega-3 fatty acid consumption is known to be beneficial for cardiometabolic health. However, the related evidence for individuals with a relatively higher intake of fish or omega-3 unsaturated fatty acids, e.g., Japanese individuals, is scarce. Therefore, this study aimed to examine the association of fish and omega-3 fatty acid intakes with the carotid intima-media thickness (C-IMT) in the Japanese population. In total, 1803 Japanese men and women aged 30–84 years without a history of myocardial infarction or angina pectoris were included in the study. The fish and omega-3 fatty acid intakes were estimated using food frequency questionnaires. The C-IMT was measured using ultrasound imaging, and the participants were classified into three groups: normal, moderate (1.1 to 1.4 mm of maximum C-IMT), and severely increased C-IMT (≥1.5 mm). Multinomial logistic regression models were used to calculate the odds ratio (OR) and 95% confidence intervals (95% CI) of the presence of moderately and severely increased C-IMT. The omega-3 fatty acid intake was shown to be associated with lower odds of severely increased C-IMT. The multivariable-adjusted OR (95%CI) was 0.55 (0.31–0.97; p for trend = 0.04). We also found a borderline significant negative association between fish intake and the presence of severely increased C-IMT. In conclusion, omega-3 fatty acid intake might protect against the development of atherosclerosis in the Japanese population.

There has been no report on the prevalence of Campylobacter spp. in farm animals in Mongolia. To uncover the prevalence of Campylobacter spp. in chickens in Mongolia and their antimicrobial resistance, in this study, we isolated and characterized Campylobacter spp. from chickens in Mongolia. We collected 71 cloacal swabs of chickens from 5 farms including 4 layer farms and one broiler farm near Ulaanbaatar city and isolated 25 Campylobacter jejuni and 6 Campylobacter coli isolates. All isolates were resistant to tetracycline, and 3 C. coli isolates were resistant to erythromycin. The C. coli isolates possessed either the erm(B) gene or nucleotide substitution at nt 2,075 of 23S rDNA, both of which are known to be associated with erythromycin resistance. Sixteen of the 31 C. jejuni/C. coli isolates (51.6%) were resistant to nalidixic acid and fluoroquinolones. All the fluoroquinolone-resistant isolates possessed amino acid substitution from threonine to isoleucine at codon 86 (nucleotide substitution: ACA to ATA). Multilocus sequence typing and phylogenetic analyses showed a variation in C. jejuni/C. coli in chickens in Mongolia. In addition, some of the C. jejuni isolates seemed to be phylogenetically close to isolates in Asian and Oceanian countries. This is the first report on the characterization of antimicrobial resistance of Campylobacter spp. in farm animals in Mongolia and is valuable for implementation of measures for a prudent use of antimicrobials in farm animals.

Geobacillus kaustophilus HTA426 is a thermophilic bacterium whose genome harbors numerous insertion sequences (IS). This study was initially conducted to generate mutant genes for thermostable T7 RNA polymerase in G. kaustophilus; however, relevant experiments unexpectedly identified that the organism transposed multiple IS elements and produced derivative cells that expressed a silent gene via transposition. The transposed elements were diverse and included members of the IS4, IS701, IS1634, and ISLre2 families. The transposition was relatively active at elevated temperatures and generated 4–9 bp of direct repeats at insertion sites. Transposition was more frequent in proliferative cells than in stationary cells but was comparable between both cells when sigX, which encodes an extra-cytoplasmic function sigma factor, was forcibly expressed. Southern blot analysis indicated that IS transposition occurred under growth inhibitory conditions by diverse stressors; however, IS transposition was not detected in cells that were cultured under growth non-inhibitory conditions. These observations suggest that G. kaustophilus enhances IS transposition via sigX-dependent stress responses when proliferative cells were prevented from active propagation. Considering Geobacillus spp. are highly adaptive bacteria that are remarkably distributed in diverse niches, it is possible that these organisms employ IS transposition for environmental adaptation via genetic diversification. Thus, this study provides new insights into adaptation strategies of Geobacillus spp. along with implications for strong codependence between mobile genetic elements and highly adaptive bacteria for stable persistence and evolutionary diversification, respectively. This is also the first report to reveal active IS elements at elevated temperatures in thermophiles and to suggest a sigma factor that governs IS transposition.

Although the pathogenesis of chronic obstructive pulmonary disease (COPD) is not yet fully understood, recent studies suggest that the disruption of the intracellular balance of oxidative (such as reactive oxygen species (ROS)) and antioxidant molecules plays an important role in COPD development and progression. Metallothionein is an endogenous metal-binding protein with reported ROS scavenging activity. Although there have been many publications on the protective effects of metallothionein in the kidney and liver, its role in COPD models such as elastase- or cigarette smoke (CS)-induced lung injury is unknown. Thus, in the present study, we analyzed the elastase-induced lung injury model using metallothionein-knockout (MT-KO; MT-1 and -2 gene deletion) mice. The expression of MT-1 and MT-2 in the lungs of MT-KO mice was markedly lower compared with that in the lungs of wildtype (WT) mice. Porcine pancreatic elastase (PPE)-induced lung injury (alveolar enlargement and respiratory impairment) was significantly exacerbated in MT-KO mice compared with WT mice. Additionally, PPE-induced increases in the number of inflammatory cells, inflammatory cytokines, and cell death in lung tissue were significantly more pronounced in MT-KO mice compared with WT mice. Finally, using an in vivo imaging system, we also found that PPE-induced ROS production in the lungs was enhanced in MT-KO mice compared with WT mice. These results suggest that metallothionein may act as an inhibitor against elastase-induced lung injury by suppressing ROS production. These results suggest that metallothionein protein, or compounds that can induce metallothionein, could be useful in the treatment of COPD.