Transcription factor GATA6 stably expressed in Chinese hamster ovary (CHO)-K1 cells is exported from the nucleus to the cytoplasm and degraded there by proteasome upon treatment with dibutylyl-cyclic AMP (dbcAMP), which is a membrane-permeable cyclic AMP (cAMP) analogue. The cAMP-dependent proteolysis of GATA6 was characterized by dissection of the GATA6 protein into a zinc-finger domain (Zf) and the surrounding region (ΔZf). These segments were separately expressed in CHO-K1 cells stably, and followed by treatment with dbcAMP. The nuclear localized Zf was degraded by proteasome similarly to the full-length GATA6. Site-directed mutants of nuclear localizing signal (NLS) (³⁴⁵RKRKPK³⁵⁰ → AAAAPK and AAAAPA) and closely related GATA4 showed the same behavior. Although nuclear-localized ΔZf was degraded by proteasome, the cytoplasmic-located ΔZf was resistant to proteolysis in contrast to the NLS mutants. We also searched for a potential NLS and nuclear export signal (NES) with computational prediction programs and compared the results with ours. All these results suggest that the amino acid sequence(s) of the Zf of GATA6 is responsive to cAMP-dependent nuclear export and proteolysis.

The extracellular signal-regulated kinase (ERK) 1 and 2 intracellular signaling pathways play key roles in a variety of cellular processes, such as proliferation and differentiation. Dysregulation of ERK1/2 signaling has been implicated in many diseases, including cancer. Although ERK1/2 signaling pathways have been extensively studied, controversy remains as to whether ERK1 and ERK2 have specific or redundant functions. In this study, we examined the functional roles of ERK1 and ERK2 in cell proliferation and cell cycle progression using an auxin-inducible degron system combined with gene knockout technology. We found that ERK1/2 double depletion, but not ERK1 or ERK2 depletion, substantially inhibited the proliferation of HCT116 cells during G1-S transition. We further demonstrated that ERK1/2-double-depleted cells were much more tolerant to etoposide-induced G2/M arrest than ERK1 or ERK2 single-knockout cells. Together, these results strongly suggest the functional redundancy of ERK1 and ERK2 in both the G1-S transition under physiological conditions and the DNA damage-induced G2/M checkpoint. Our findings substantially advance understanding of the ERK1/2 pathways, which could have strong implications for future pharmacological developments.

Increasing incidences of resistance to antibiotics by pathogenic bacteria is a worldwide concern and isolation of antibiotic-resistant strains of Niallia circulans (formerly known as Bacillus circulans), an opportunistic human pathogen, has been reported. Due to their lack of ethical constraints as well as their cost-effective rearing, invertebrates have been commonly used to study infection by bacteria pathogenic to humans. In this study, we demonstrate that a foodborne strain of N. circulans kills larvae of the silkworm, Bombyx mori within 48 h after hemolymph injection. The infected larvae turned black with an increase in the phenoloxidase (PO) activity in the hemolymph. Midgut injection of N. circulans resulted in the killing of larvae within 96 h. A significant increase in bacterial load was observed in the hemolymph 12 h after infection. The viable hemocyte number decreased to 48% within 12 h of injection. RT-qPCR analysis revealed that upon hemolymph infection with N. circulans the expression of the antimicrobial peptide (AMP) genes, Bmdefensin-B and Bmgloverin-3, were upregulated 2.5- and 1.8-fold, respectively, whereas 1.6-fold upregulation was observed for BmToll-2 in the larval fat body. Therapeutic effects of antibiotics like tetracycline, imipenem, ceftriaxone, ampicillin, and clindamycin were observed against N. circulans in the Bombyx larvae with varying efficacies. Results from this study suggest that larvae of B. mori can be used as infection models for screening therapeutics that are effective against N. circulans.

Cervical lymph node metastasis (CLNM) of papillary thyroid carcinoma (PTC) is directly associated with clinical management and prognosis. In this study, we aimed to evaluate the value of conventional ultrasound (US) combined with ENST00000438158 in predicting CLNM of PTC. Fourty-nine PTC patients underwent US examination and US-guided fine needle aspiration (FNA). ENST00000438158 expression in FNA cytological specimens and PTC cell lines was detected using real-time reverse transcription polymerase chain reaction (qRT-PCR). The role of ENST00000438158 expression in the proliferation, migration, invasion, apoptosis, and cell cycle of PTC cells was investigated by Cell Counting Kit-8 (CCK8) and clone formation experiments, transwell assay, and flow cytometry, respectively. Calcification, capsule contact, and low ENST00000438158 expression were independently associated with PTC with CLNM (all p < 0.05). The combination of multiple US features was more valuable than a single US feature in predicting CLNM in PTC. Adding ENST0000438158 to US greatly improved the value of differentiation of PTC with or without CLNM. In conclusion, ENST00000438158 is a potential molecular marker for predicting CLNM in PTC. ENST00000438158 combined with US features is highly valuable for predicting CLNM in PTC.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a serious threat to global development. Rapid and accurate diagnosis is critical for containing the pandemic and treating patients in time. As the gold standard for SARS-CoV-2 diagnosis, the qualitative reverse transcription-PCR (RT-qPCR) test has long been criticized for its long detection time. In this study, we optimized the primers and probes targeting SARS-CoV-2 ORF1ab and N gene designed by the Chinese Center for Disease Control and Preventions (CDC) to increase their Tm values to meet the optimal elongation temperature of Taq DNA polymerase, thus greatly shortened the elongation time. The higher elongation temperature in turn narrowed the temperature range of the reaction and saved more time. In addition, by shortening the distance between the fluorophore at the 5' end and the quencher in the middle we got a probe with higher signal-to-noise ratio. Finally, by using all these measures and optimized RT-qPCR program we successfully reduced the time (nucleic acid extraction step is not included) for nucleic acid test from 74 min to 26 min.

During the coronavirus disease 2019 (COVID-19) outbreak, firefighters have been working in an environment that is both physically and mentally taxing. This study aimed to investigate factors affecting health-related quality of life (HRQOL) among firefighters in Japan during the COVID-19 pandemic. A total of 227 firefighters from a single firefighting organization were surveyed in June 2021, during the fourth infection spread period of COVID-19 in Japan. Regression analysis was performed to examine factors affecting HRQOL of firefighters measured with the SF-8. In the present study, factors affecting HRQOL among firefighters during the COVID-19 pandemic were lack of sleep, physical abnormalities due to infection control measures, exercise habits, living with family members, and history of suspected COVID-19 infection. The present findings may help develop support services for first responders, including firefighters during the COVID-19 pandemic.

Intravenous infusion using a peripheral intravenous catheter (PIVC) is often complicated by catheter failure (CF). We hypothesized that catheterization of an upper arm vein instead of a forearm vein may help prevent CF. This study was designed to compare the incidence of CF in patients receiving hyper-stimulant drugs when catheters are placed in the forearm using short PIVCs (SPCs) with that when catheters are placed in the upper arm using the new long PIVCs. Patients admitted to a university hospital in Tokyo, Japan were enrolled in this study and were assigned to the SPC or the new long PIVC group. The primary outcome was the incidence of CF until 7 days. The secondary outcomes were the number of CFs per 1,000 days, the duration of the indwelling catheter, and the presence of thrombi and subcutaneous edema. Forty-seven patients were analyzed (median age, 67.0 years). The incidence of CF was 0% in the new long PIVCs and 32.0% (8 catheters) in the SPCs (p = 0.007), and the number of CF per 1,000 days was 0/1,000 and 81.7/1,000 days, respectively (p = 0.001). A significant difference in the duration of the indwelling catheter until CF occurrence was observed between the two groups (p = 0.004). Thrombi and subcutaneous edema were observed more frequently in the SPC group (p < 0.001). Catheterization of an upper arm vein using the new long PIVC to administer a hyper-stimulant drug might reduce CF compared with catheterization of a forearm vein using SPC.

The present retrospective study aimed to examine the real-world data regarding time-dependent changes in the age distribution of patients with coronavirus disease 2019 (COVID-19) as well as the severity and infectivity in a regional core hospital in Japan. Patients with COVID-19 who visited the fever outpatient branch in Takagi Hospital during phase I (May 1 to December 31, 2021), and during phase II (January 1 to April 30, 2022) were evaluated. The age distribution of outpatients and the characteristics of inpatients aged > 75 years were compared between phases I and II. The age distribution of outpatients shifted from the older generation in phase I to the younger generation in phase II (p < 0.01). Disease severity might be reduced in a time-dependent manner with a decrease in the hospitalization rate (phase I: 145/368 (39.4%); phase II: 104/1496 (7.0%); p < 0.01) and mortality rate (phase I: 10/368 (2.7%); phase II: 7/1496 (0.5%); p < 0.01). The number of patients increased in phase II (374.0/month) compared to that in phase I (36.8/month). Regarding the older inpatients, the disease severity of COVID-19 and hospitalization days were reduced in phase II compared to those in phase I (p < 0.01, each). In conclusion, the present study suggests a change in the age distribution of patients with COVID-19, a decrease in toxicity, and an increase in infectivity of severe acute respiratory syndrome coronavirus 2 in a time-dependent manner.

Various studies have reported relationships between salt intake and diseases, such as hypertension, cardiovascular disease, stroke, gastric cancer, and bronchial asthma. However, no reports exist on the relationship between salt intake and food allergies. In this study, we investigated the effect of continuous ingestion of sodium chloride (NaCl) on allergy symptoms using a mouse model of food allergy. BALB/c mice were divided into four groups of 6-8 animals each. The control-water group (CW) and sensitization-water group (SW) groups were provided free access to water, and the control-1% NaCl group (CS) and sensitization-1% NaCl group (SS) groups were provided a 1% NaCl solution. The SW and SS groups were sensitized with 50 µg ovalbumin (OVA) at 2 timepoints by intraperitoneal injection. After oral administration of OVA, anaphylactic response was measured and blood was collected. The mice were sacrificed, and serum levels of OVA and anti-OVA immunoglobulin (Ig)E and IgG1 were measured by enzyme-linked immunosorbent assays. The sodium ion (Na⁺) concentrations in duodenal and jejunal extracts were measured using a Na⁺ ion meter. The results suggested that continuous ingestion of a 1% NaCl solution for 36 days promoted allergen absorption and may have aggravated allergy symptoms in the mice. However, NaCl ingestion did not affect Na⁺ concentrations in the small intestine or OVA-specific antibody production.

We studied 95 patients with infantile hemangioma (IH) treated with propranolol at the Department of Dermatology, Kumamoto University Hospital, from November 2016 to January 2022, based on sex, site, clinical classification, duration of treatment, and residual lesions after treatment. Four of the 95 patients discontinued propranolol due to side effects, and 55 completed follow-ups at our hospital. We observed that 30.1% showed complete resolution of the skin rash, while the remaining 69.8% had erythema or atrophic scarring. Complete resolution occurred in 70% of the cases with the subcutaneous type but only in 15% with the tumor type. Seventeen of the 55 patients who completed follow-ups were treated with propranolol combined with laser therapy. Combined use of propranolol and laser therapy significantly reduced severe erythema compared to the propranolol monotherapy. These results suggest that propranolol therapy in IH often leaves erythema except in the subcutaneous type and that an improvement in erythema can be expected when propranolol is combined with laser therapy.