Effect of triclocarban on membrane potential of rat thymocytes : Assessment with bis-(1,3-dibutylbarbituric acid)trimethine oxonol

The effect of triclocarban (TCC), an　environmental pollutant from household items and health care products, on membrane potential of rat thymocytes was examined by a flow cytometry with a fluorescent probe sensitive to membrane potential, bis-(1,3-dibutylbarbituric acid)trimethine oxonol, because TCC changes intracellular ionic circumstance that may affect the membrane potential. TCC at 0.3 μM or more (up to 3 μM) depolarized the membranes. This TCC-induced phenomenon was against our prediction because TCC increases intracellular Ca2+ concentration that activates Ca2+-dependent K+ channels, resulting in a hyperpolarization. The depolarization was still observed under Ca2+-free condition, but not under Na+-free condition. Furthermore, TCC hyperpolarized the membranes under Ca2+- and Na+-free condition. To see if TCC inhibits Ca2+-dependent hyperpolarization, the effect of A23187, a calcium ionophore, on the membrane potential was examined in the cells treated with TCC. A23187 induced large depolarization in the cells treated with 0.3–3 μM TCC. The A23187-induced depolarization in the presence of TCC was greatly attenuated under Na+-free or Ca2+-free condition whereas A23187 elicited hyperpolarization in the cells treated with 0.3–3 μM TCC under Ca2+- and Na+-free condition. Results suggest that 0.3–3 μM TCC increases membrane permeability of Na+ and Ca2+, resulting in the depolarization. Large depolarization induced by TCC in the presence of external Ca2+ and Na+ may mask the hyperpolarization elicited via the increase in intracellular Ca2+ concentration by TCC. Thus, there is a possibility that TCC depolarizes membranes of lymphocytes, resulting in alteration of cellular functions of lymphocytes

Fibrosis in metastatic lymph nodes is clinically correlated to poor prognosis in colorectal cancer.

Background:Tumor microenvironment including fibrosis has a pivotal role in cancer growth and distant metastasis. Fibrosis is a known risk factor for carcinogenesis, but its biological role in disease invasion and metastasis in colorectal cancer (CRC) remains unclear. In particular, there is no report on how fibrosis of metastatic lymph nodes(MLNs) in CRC contributes to prognosis.Methods:We reviewed 94 colorectal adenocarcinoma patients with MLNs who underwentcolectomy. Both the primary tumors and MLNs were analyzed for alpha-smooth muscleactin (α-SMA) expression and collagen deposition.Results:Higher α-SMA expression and collagen deposition in MLNs were associated withsignificantly shorter relapse-free survival and overall survival in CRC patients. α-SMA expression in MLNs (HR, 1.53; p = 0.034) was independent predictive factor of overallsurvival in multivariate Cox proportional hazards regression analysis of clinicopathologicalfactors. In the Stage III patient subgroup, α-SMA expression in MLNs was a strong prognostic marker (HR, 3.01; p = 0.006). On the other hand, higher α-SMA expressionand collagen deposition in primary tumors were associated with short overall survival, but they were not significant factors in multivariate Cox regression analyses. In MLNs,the podoplanin signals co-localized with α-SMA expression and were confirmed by thedual immunofluorescence staining, implying that the MLN stromal cells were fibroblasticreticular cells.Conclusion:Both high collagen deposition and high α-SMA expression in MLNs predicted poor prognosis in CRC

DOCK2 is involved in the host genetics and biology of severe COVID-19

「コロナ制圧タスクフォース」COVID-19疾患感受性遺伝子DOCK2の重症化機序を解明 --アジア最大のバイオレポジトリーでCOVID-19の治療標的を発見--. 京都大学プレスリリース. 2022-08-10.Identifying the host genetic factors underlying severe COVID-19 is an emerging challenge. Here we conducted a genome-wide association study (GWAS) involving 2, 393 cases of COVID-19 in a cohort of Japanese individuals collected during the initial waves of the pandemic, with 3, 289 unaffected controls. We identified a variant on chromosome 5 at 5q35 (rs60200309-A), close to the dedicator of cytokinesis 2 gene (DOCK2), which was associated with severe COVID-19 in patients less than 65 years of age. This risk allele was prevalent in East Asian individuals but rare in Europeans, highlighting the value of genome-wide association studies in non-European populations. RNA-sequencing analysis of 473 bulk peripheral blood samples identified decreased expression of DOCK2 associated with the risk allele in these younger patients. DOCK2 expression was suppressed in patients with severe cases of COVID-19. Single-cell RNA-sequencing analysis (n = 61 individuals) identified cell-type-specific downregulation of DOCK2 and a COVID-19-specific decreasing effect of the risk allele on DOCK2 expression in non-classical monocytes. Immunohistochemistry of lung specimens from patients with severe COVID-19 pneumonia showed suppressed DOCK2 expression. Moreover, inhibition of DOCK2 function with CPYPP increased the severity of pneumonia in a Syrian hamster model of SARS-CoV-2 infection, characterized by weight loss, lung oedema, enhanced viral loads, impaired macrophage recruitment and dysregulated type I interferon responses. We conclude that DOCK2 has an important role in the host immune response to SARS-CoV-2 infection and the development of severe COVID-19, and could be further explored as a potential biomarker and/or therapeutic target

Transudative pleural effusion in pleuritis associated with immunoglobulin G4‐related disease diagnosed by thoracoscopy under local anaesthesia

Abstract Immunoglobulin G4 (IgG4)‐related disease is a chronic inflammatory condition often characterized by exudative pleural effusions. However, transudative pleural effusions, like in the presented case of an 80‐year‐old man with multiple comorbidities, are less common but possible. Despite initial treatment with diuretics, the effusion persisted, prompting further investigation. Medical thoracoscopy revealed lymphatic follicle hyperplasia and an abundance of IgG4‐positive plasmacytoid cells, confirming IgG4‐related pleuritis. This case underscores the importance of considering inflammatory etiologies, such as IgG4‐related disease, when faced with unresponsive transudative pleural effusions. Thoracoscopy serves as a valuable diagnostic tool in such scenarios, allowing for precise diagnosis and appropriate management

Robot-assisted radical prostatectomy significantly reduced biochemical recurrence compared to retro pubic radical prostatectomy

Abstract Background The pathological and oncological outcomes of retro-pubic radical prostatectomy (RRP) and robot-assisted radical prostatectomy (RARP) have not been sufficiently investigated. Methods Treatment-naïve patients with localized prostate cancer (PC) (n = 908; RRP, n = 490; and RARP, n = 418) were enrolled in the study. The clinicopathological outcomes, rate and localization of the positive surgical margin (PSM), localization of PSM, and biochemical recurrence (BCR)-free survival groups were compared between RRP and RARP. Results The median patient age and serum PSA level (ng/mL) at diagnosis were 67 years and 7.9 ng/ml, respectively, for RRP, and 67 years and 7.6 ng/ml, respectively, for RARP. The overall PSM rate with RARP was 21%, which was 11% for pT2a, 12% for pT2b, 9.8% for pT2c, 43% for pT3a, 55% for pT3b, and 0% for pT4. The overall PSM rate with RRP was 44%, which was 12% for pT2a, 18% for pT2b, 43% for pT2c, 78% for pT3a, 50% for pT3b, and 40% for pT4. The PSM rate was significantly lower for RARP in men with pT2c and pT3a (p < 0.0001 for both). Multivariate analysis showed that RARP reduced the risk of BCR (hazard ratio; 0.6, p = 0.009). Conclusions RARP versus RRP is associated with an improved PSM rate and BCR. To examine the cancer-specific survival, further investigations are needed

Bordetella Dermonecrotic Toxin Is a Neurotropic Virulence Factor That Uses CaV3.1 as the Cell Surface Receptor

Bordetella pertussis, which causes pertussis, a contagious respiratory disease, produces three major protein toxins, pertussis toxin, adenylate cyclase toxin, and dermonecrotic toxin (DNT), for which molecular actions have been elucidated. The former two toxins are known to be involved in the emergence of some clinical symptoms and/or contribute to the establishment of bacterial infection. In contrast, the role of DNT in pertussis remains unclear. Our study shows that DNT affects neural cells through specific binding to the T-type voltage-gated Ca2+ channel that is highly expressed in the central nervous system and leads to neurological disorders in mice after intracerebral injection. These data raise the possibility of DNT as an etiological agent for pertussis encephalopathy, a severe complication of B. pertussis infection.Dermonecrotic toxin (DNT) is one of the representative toxins produced by Bordetella pertussis, but its role in pertussis, B. pertussis infection, remains unknown. In this study, we identified the T-type voltage-gated Ca2+ channel CaV3.1 as the DNT receptor by CRISPR-Cas9-based genome-wide screening. As CaV3.1 is highly expressed in the nervous system, the neurotoxicity of DNT was examined. DNT affected cultured neural cells and caused flaccid paralysis in mice after intracerebral injection. No neurological symptoms were observed by intracerebral injection with the other major virulence factors of the organisms, pertussis toxin and adenylate cyclase toxin. These results indicate that DNT has aspects of the neurotropic virulence factor of B. pertussis. The possibility of the involvement of DNT in encephalopathy, which is a complication of pertussis, is also discussed