Response to dabrafenib plus trametinib on a rare BRAF mutation (V600_W604 deletion-insertion R) in an advanced non-small cell lung cancer patient.

Although dabrafenib plus trametinib has been approved for BRAF V600E mutation positive advanced non-small cell lung cancer (NSCLC), data on its efficacy against uncommon BRAF mutations are still limited due to their rare frequency. We report a case of 70-year-old woman with BRAF V600_W604 deletion-insertion R-positive stage IVA lung adenocarcinoma, who was successfully treated with dabrafenib plus trametinib. Herein, we discuss the oncogenic role of uncommon BRAF mutations and highlight the importance of performing comprehensive genomic profiling on patients without any targetable gene alterations in companion diagnostics.

A case of eosinophilic bronchiolitis after the initiation of immune checkpoint inhibitor.

A 50-year-old Japanese woman with advanced breast cancer presented with productive cough and dyspnea while she was receiving a sixth cycle of chemotherapy including atezolizumab. Chest computed tomography revealed bronchiolitis and transbronchial lung cryobiopsy revealed eosinophilic bronchiolitis. Corticosteroid therapy successfully resolved her symptoms. Eosinophilic bronchiolitis is a rare but important immune-related adverse event; herein, we discuss its diagnosis and possible pathophysiology.

Zranb1-mutant mice display abnormal colonic mucus production and exacerbation of DSS-induced colitis.

Expression of mucin MUC2, a component of the colonic mucus layer, plays a crucial role in intestinal homeostasis. Here, we describe a new regulator of MUC2 expression, the deubiquitinase ZRANB1 (Trabid). A ZRANB1 mutation changing cysteine to serine in amino acid position 443, affects ubiquitination. To analyze ZRANB1 function in the intestine, we generated Zranb1 C443S mutant knock-in (Zranb1C443S/C443S) mice using the CRISPR/Cas9 system. Zranb1C443S/C443S mice exhibited decreased mRNA expression and MUC2 production. Colonic organoids from Zranb1C443S/C443S mice displayed decreased Muc2 mRNA expression following differentiation into goblet cells. Finally, we analyzed dextran sulfate sodium-induced colitis to understand ZRANB1's role in intestinal inflammation. Zranb1C443S/C443S mice with colitis exhibited significant weight loss, reduced colon length, and worsening clinical and pathological scores, indicating that ZRANB1 contributes to intestinal homeostasis. Together, these results suggest that ZRANB1 regulates MUC2 expression and intestinal inflammation, which may help elucidating the pathogenesis of inflammatory bowel disease and developing new therapeutics targeting ZRANB1.

Progressive pulmonary fibrosis due to diffuse alveolar damage in a COVID-19-infected autopsy case.

We encountered a patient with severe coronavirus disease 2019 (COVID-19)-related pneumonia, who died of progressive respiratory acidosis after 2 months of treatment with mechanical ventilation. The autopsy revealed diffuse alveolar damage (DAD) without any active signs of fungal or bacterial infections, suggesting prolonged and over-activated immune responses against COVID-19 infection. When COVID-19 patients develop acute respiratory distress syndrome, it is essential to remember that the infection can progress to DAD a few months after the disease onset.

Nickel particles are present in Crohn's disease tissue and exacerbate intestinal inflammation in IBD susceptible mice.

Crohn's disease is an inflammatory disease of the gut caused by a complex interplay among genetic, microbial, and environmental factors. The intestinal tract is constantly exposed to metals and other trace elements ingested as food. Synchrotron radiation-induced X-ray fluorescence spectroscopy and X-ray absorption fine structure analysis revealed the deposition of nickel particles within Crohn's disease tissue specimens. After nickel particle stimulation, THP-1 cells showed filopodia formation and autophagic vacuoles containing lipid bodies. Nickel particles precipitated colitis in mice bearing mutations of the IBD susceptibility protein A20/TNFAIP3. Nickel particles also exacerbated dextran sulfate sodium-induced colitis in mice harboring myeloid cell-specific Atg5 deficiency. These findings illustrate that nickel particle ingestion may worsen Crohn's disease by perturbing autophagic processes in the intestine, providing new insights into environmental factors in Crohn's disease pathogenesis.

An NEFH founder mutation causes broad phenotypic spectrum in multiple Japanese families.

BACKGROUND AND AIMS: Mutations in neurofilament genes have been linked to several neuromuscular disorders. The neurofilament heavy (NEFH) gene was identified as the causative gene of Charcot-Marie-Tooth disease type 2CC (CMT2CC) in 2016, with a toxic gain of function mechanism caused by the translation and aggregation of cryptic amyloidogenic element (CAE) in the 3' untranslated region (UTR). But the NEFH-related clinical and genetic spectrums are still unclear in Japan. METHODS: We analyzed all variants in the NEFH gene from our in-house whole-exome sequencing data, established from Japanese nationwide patients with neuromuscular disorders, including Charcot-Marie-Tooth (CMT) disease and spinal muscular atrophy (SMA). RESULTS: We identified a c.3017dup (p.Pro1007Alafs*56) variant in NEFH from three families clinically diagnosed with CMT, and one family with SMA. In addition to the patients presented with typical peripheral neuropathies, pyramidal signs were observed from one CMT patient. Whereas the SMA patients showed severe characteristic weakness of triceps brachii and quadriceps femoris. All of these four families reside in Kagoshima Prefecture of Japan, and a following haplotype analysis strongly suggests a founder effect. INTERPRETATION: This is the original report referring to a founder mutation in NEFH. The clinical diversity in our study, comprising CMT, with or without pyramidal signs, and SMA, suggest an extensive involvement of peripheral nerve, anterior horn cells, or both. Our findings broaden the phenotypic spectrum of NEFH-related disorders.

Isolation of Patients With Bacterial Pneumonia Suspected of COVID-19 Leads to Prolonged Hospitalization.

Objective The coronavirus disease 2019 (COVID-19) pandemic has changed the inpatient treatment practice for bacterial pneumonia. Upon hospitalization, isolation is required while waiting for the polymerase chain reaction (PCR) test result, which may lead to limited access to medical resources and fewer room visits by medical staff. However, little is known about the relationship between isolation and the clinical outcomes of bacterial pneumonia. We hypothesized that isolation of suspected COVID-19 patients who are eventually diagnosed with bacterial pneumonia is associated with a prolonged length of hospitalization as compared with non-isolation. Patients This is a single-center, retrospective observational study of hospitalized adult patients diagnosed with bacterial pneumonia from January 2018 to September 2021. The patients were divided into a non-isolated (patients hospitalized between January 2018 and December 2019, who were not isolated at all) and an isolated group (patients hospitalized between January 2020 and September 2021, who were initially isolated because COVID-19 was suspected). The primary outcome was a prolonged length of hospitalization (≥14 days), and its relationship with isolation was analyzed using logistic regression analysis adjusted for age, sex, CURB-65, and the Charlson Comorbidity Index. Results Among 917 eligible patients, 214 (23%) underwent isolation. In the logistic regression analysis, the isolated group independently had a prolonged length of hospitalization as compared with the non-isolated group (odds ratio 1.49; 95% confidence interval 1.08-2.07, p=0.015). There was no significant difference in antibiotic use duration between the groups. Conclusion The isolation of bacterial pneumonia patients suspected of COVID-19 was associated with prolonged length of hospitalization.

Characteristics of systemic infection and host responses in chickens experimentally infected with Salmonella enterica serovar Gallinarum biovar Gallinarum.

Salmonella enterica serovar Gallinarum biovar Gallinarum (S. Gallinarum) is a host-specific pathogen causing systemic infection in poultry, which leads to significant economic losses due to high mortality. However, little is known about the dynamic process of systemic infection and pathogenic characteristics of S. Gallinarum in chickens. In the present study, we developed an oral infection model that reproduces the pathology of S. Gallinarum and clarified the host immune response of the infected chickens. Chickens at 20 days of age orally inoculated at a dose of 108 colony forming unit (CFU) showed typical clinical signs of fowl typhoid and died between 6 and 10 days post infection. The inoculated S. Gallinarum rapidly disseminated to multple organs and the bacterial counts increased in the liver and spleen at 3 days post infection. Pathological changes associated wirh inflammation in the liver and spleen became apparent at 4 days post infection, and increased expression of interferon (IFN)-γ and interleuikin (IL)-12 in the liver and spleen did not observed until 3 days post infection. These results indicate that S. Gallinarum rapidly spread to entire body through intestine, and the low-level of inflammatory responses in the liver during the early stage of infection may contribute to rapid, systemic dissemination of the bacteria. Our infection model and findings will contribute to the better understanding of the pathogenic mechanism of S. Gallinarum, and provide new insights into the prevention and control of fowl typhoid.

Nickel ions attenuate autophagy flux and induce transglutaminase 2 (TG2) mediated post-translational modification of SQSTM1/p62.

Nickel, the most frequent contact allergy cause, is widely used for various metallic materials and medical devices. Autophagy is an intracellular protein degradation system and contributes to metal recycling. However, it is unclear the functions of nickel in autophagy. We here demonstrated that NiCl2 induced microtubule-associated protein 1 light chain 3 (LC3)-II and LC3 puncta, markers of autophagosomes. Bafilomycin A1 (BafA1) treatment did not enhance LC3 puncta under NiCl2 stimulation, suggesting that NiCl2 did not induce autophagic flux. In addition, NiCl2 promotes the accumulation of SQSTM1/p62 and increased SQSTM1/p62 colocalization with lysosomal-associated membrane protein 1 (LAMP1). These data indicated that NiCl2 attenuates autophagic flux. Interestingly, NiCl2 induced the expression of the high-molecular-weight (MW) form of SQSTM1/p62. Inhibition of NiCl2-induced reactive oxygen species (ROS) reduced the high-MW SQSTM1/p62. We also showed that NiCl2-induced ROS activate transglutaminase (TG) activity. We found that transglutaminase 2 (TG2) inhibition reduced high-MW SQSTM1/p62 and SQSTM1/p62 puncta under NiCl2 stimulation, indicating that TG2 regulates SQSTM1/p62 protein homeostasis under NiCl2 stimulation. Our study demonstrated that nickel ion regulates autophagy flux and TG2 restricted nickel-dependent proteostasis.

Effects of metformin on the prevention of bisphosphonate-related osteonecrosis of the jaw-like lesions in rats.

PURPOSE: In this study, we aimed to investigate the effect of glucose metabolism on bone healing after tooth extraction in an osteoporosis rat model administered zoledronic acid (ZA) and dexamethasone (DX). METHODS: In total, 24 male Wistar rats (4 weeks old) were randomly assigned to four groups: Control (subcutaneous physiological saline), ZD (subcutaneous ZA and DX twice a week), Ins+ZD (subcutaneous insulin followed by ZD treatment), and Met+ZD (oral metformin followed by ZD treatment). Blood was collected every two weeks . Two weeks after treatment initiation, the first molar tooth on the right maxilla was extracted from all rats. Four weeks later, the rats were sacrificed, and bone healing was assessed. Maxillae samples were fixed and scanned using micro-computed tomography for quantifying areas of bone defects. Hematoxylin-eosin and tartrate-resistant acid phosphatase (TRAP) staining were performed to evaluate bone apoptosis and osteoclast number. RESULTS: In all experimental groups, body weight was statistically lower than that in the Control group, with no changes observed in uncarboxylated osteocalcin concentrations. The radiological analysis revealed that insulin or metformin administration improved healing in the tooth extraction socket (p < 0.01). Histological examination revealed that the osteonecrosis area was reduced in the Ins+ZD and Met+ZD groups (p < 0.01). TRAP staining presented increased osteoclast numbers in the ZD group when compared with that observed in the Control. CONCLUSIONS: Tooth extraction with long-term ZA and DX administration inhibited bone remodeling and induced bisphosphonate-related osteonecrosis of the jaw-like lesions. Metformin exerted protective effects ag ainst osteonecrosis of the jaw.

Course of the thoracic nerves around the umbilicus within the posterior layer of the rectus sheath: a cadaver study.

Rectus sheath block is used to anesthetize thoracic nerves around the umbilicus. However, the appropriate point for anesthetic injection during rectus sheath block has not been determined anatomically. Here, we examined the course of thoracic nerve T10 at the posterior layer of the rectus sheath and the anatomical relationship between the nerve and the rectus abdominis and transversus abdominis muscles in formalin-fixed adult cadavers. The cranio-caudal distance from a horizontal line running through the umbilicus to where the thoracic nerve T10 passes through the posterior layer of the rectus sheath was 33.8 ± 14.4 (mean ± standard deviation) mm, while that from the horizontal line running through the umbilicus to the position where the lateral edge of the rectus abdominis muscle and the medial border of the transversus abdominis muscle cross was 33.1 ± 17.1 mm. The position where the lateral edge of the rectus abdominis muscle and the medial border of the transversus abdominis muscle cross approximates the position where thoracic nerves T10 passes through the posterior layer of the rectus sheath. Our results identify effective landmarks to guide the performance of rectus sheath block.

Receptor-Interacting Protein Kinase 3 (RIPK3) inhibits autophagic flux during necroptosis in intestinal epithelial cells.

Autophagy is an intracellular process that regulates the degradation of cytosolic proteins and organelles. Dying cells often accumulate autophagosomes. However, the mechanisms by which necroptotic stimulation induces autophagosomes are not defined. Here, we demonstrate that the activation of necroptosis with TNF-α plus the cell-permeable pan-caspase inhibitor Z-VAD induces LC3-II and LC3 puncta, markers of autophagosomes, via the receptor-interacting protein kinase 3 (RIPK3) in intestinal epithelial cells. Surprisingly, necroptotic stimulation reduces autophagic activity, as evidenced by enlarged puncta of the autophagic substrate SQSTM1/p62 and its increased colocalization with LC3. However, necroptotic stimulation does not induce the lysosomal-associated membrane protein 1 (LAMP1) nor syntaxin 17, which mediates autophagosome-lysosome fusion, to colocalize with LC3. These data indicate that necroptosis attenuates autophagic flux before the lysosome fusion step. Our findings may provide insights into human diseases involving necroptosis.

The effects of hyperglycaemia on peri-implant tissues after osseointegration.

PURPOSE: We assessed the effects of hyperglycaemia induced by streptozotocin (STZ) on peri-implantitis developing after implant osseointegration. METHODS: Thirty-six male Wistar rats (4 weeks old) were used. We placed titanium implants 4 weeks after extraction of the maxillary first molars. Healing abutments were attached 4 weeks later. After osseointegration was confirmed, the rats were divided into control, hyperglycaemia (STZ), and STZ with insulin (STZ+INS) groups. Hyperglycaemia was induced by a single injection of 50mg/kg STZ. Silk ligatures were placed on only the right sides (i.e. ligature sides), not on the left sides. Peri-implant tissues extracted at 4 weeks post-ligation were analysed both radiologically (via micro-computed tomography) and histologically (via toluidine blue staining). Total RNA was also extracted and analysed by quantitative PCR to detect TNF-α, IL-1ß and the receptor of advanced glycation end products (RAGE). Additionally, advanced glycation end products (AGEs) were measured by ELISA. RESULTS: Radiological and histological analyses showed that bone loss on the non-ligature sides was significantly greater in the STZ than the control group. However, on the ligature sides, bone loss was greater than on the non-ligature sides, and no significant difference was evident among the three groups. The levels of mRNAs encoding TNF-α, IL-1ß, RAGE, and AGEs on the ligature sides were significantly upregulated (all P<0.05) in the STZ compared to the control group. CONCLUSIONS: Although hyperglycaemia could be associated with bone loss around implants with increased AGE production and RAGE expression, hyperglycaemia does not become a triggering factor of ligature induced peri-implantitis.

In Vitro Differentiation of Chicken Astrocytes: Growth, Morphology, and Protein Expression of Astrocytes in Primary Cultures.

Astrocytes regulate synaptic transmission in the central nervous system. Astrocytes in vivo have "stems" that express glial fibrillary acidic protein (GFAP), intermediate filaments, and peripheral astrocyte processes (PAPs), which contain actin-rich cytoskeletal structures. At the PAPs, the perisynaptic glia contacts and enwraps synapses, and modulates glia-neuronal communication. Cultured astrocytes have been an invaluable tool for studying roles of astrocytes; however, the morphology of mammalian primary astrocytes cultured in conventional medium containing fetal bovine serum (FBS) was similar to that of fibroblasts, and many culture conditions have been developed to generate stellate astrocytes observed in vivo. Avian astrocytes have been prepared from embryonic chick forebrain and maintained at a high cell density in conventional FBS-containing medium as mammalian astrocytes, thus the morphological analysis of chicken astrocytes has not yet been performed. In the present study, we report that the morphology of astrocytes freshly harvested from the forebrain of a chicken embryo in serum-free Neurobasal medium with B-27 supplement and basic fibroblast growth factor (bFGF) is similar to that of the astrocyte morphology in vivo. We also find that astrocytes in this medium express similar levels of GFAP and two actin-binding proteins as astrocytes in conventional FBS-containing medium, although they have different morphologies. Furthermore, we confirmed that cryopreserved astrocytes differentiate faster than freshly harvested astrocytes.

Implant-supported fixed prosthesis improves nutrient intake in patients with partial edentulous posterior regions.

PURPOSE: This study investigated changes in food and nutrient intake after implant-supported fixed prosthesis treatment in patients with partial edentulous posterior regions. METHODS: This study included 30 patients who received implant treatment with fixed prostheses in the posterior region. Food and nutrient intake was evaluated using a brief self-administered diet history questionnaire at baseline and post-implant treatment, and the results were statistically analyzed. RESULTS: Treatment with implant-supported fixed prostheses in patients with posterior edentulous conditions tended to increase the amounts of soy products and vegetables consumed: in particular, intake of carrot and squash was significantly increased. The total energy, protein, lipid, and carbohydrate intakes were comparable between baseline and post-implant treatment. On the other hand, the vegetable protein, α-carotene, daidzein, and genistein intakes were significantly increased, and dietary fiber and ß-carotene intakes tended to be increased in patients with implant-supported fixed prostheses. CONCLUSIONS: Implant-supported fixed prostheses in patients with posterior edentulous conditions affected food intake, resulting in improved nutrient intake.

Efficacy of electric-powered cleaning instruments in edentulous patients with implant-supported full-arch fixed prostheses: a crossover design.

BACKGROUND: The aim of this study was to evaluate the plaque removal efficacies of electric toothbrushes and electric dental floss compared with conventional manual toothbrushing in cleaning the fitting surface of an All-on-4™ concept (Nobel Biocare, Zürich-Flughafen, Switzerland) implant-supported fixed dental prosthesis (FDP). METHODS: Nine patients with maxillary edentulous arches participated in the study. We investigated two electric-powered brushes (Sonicare Diamond Clean®, Koninklijke Philips N.V., Amsterdam, the Netherlands [SD group], and the Oral-B Professional Care Smart Series 5000®, Braun GmbH, Kronberg, Germany [OralB group]) and one electric dental floss unit (Air Floss®, Koninklijke Philips N.V. [AF group]). A manual toothbrush (Tuft24® MS, OralCare Inc., Tokyo, Japan) was used by the control group. The fitting surface of the FDP was stained to allow visualization of the entire accumulated plaque area. Both the buccal and palatal portions of the plaque area were assessed before and after brushing to evaluate each instrument's plaque removal rate using a crossover study design. Two-week washout periods were employed between each evaluation. RESULTS: The plaque removal rates were 53.5 ± 8.5%, 70.9 ± 6.5%, 75.4 ± 6.3%, and 74.4 ± 4.2% for the control, AF, OralB, and SD groups, respectively. When participants were divided into two groups based on their plaque removal rates with a manual toothbrush (poor brushing and good brushing), the poor brushing group showed significant improvement in the plaque removal rate when using electric-powered toothbrushes. The plaque removal rates for the buccal area were significantly higher for the OralB and SD groups than for the manual brushing group (control group), with rates of 52.8 ± 7.9%, 70.1 ± 7.3%, 77.7 ± 6.5%, and 79.5 ± 3.7% for the control, AF, OralB, and SD groups, respectively. The plaque removal rates in the palatal area were consistently lower than those in the buccal area for each of the three electric instruments. CONCLUSIONS: The results suggest that patients who are not adept at manual toothbrushing may potentially improve their removal of plaque from the fitting surfaces of FDPs by using electric toothbrushes.

Ten-year survival of immediate-loading implants in fully edentulous mandibles in the Japanese population: a multilevel analysis.

PURPOSE: To evaluate the long-term clinical results of and risk factors for immediate-loading implant treatment of completely edentulous mandibles. METHODS: We retrospectively studied 220 implants in 52 patients who received immediate-loading implants in fully edentulous mandibles. Kaplan-Meier survival analyses, log-rank tests, and multilevel mixed-effects parametric survival analysis was used for statistical analyses. RESULTS: Thirteen of implants in seven patients failed, and the 10-year cumulative implant survival rate was 93.9 % and significantly (p=0.049) higher in women than in men. None of the predictor variables were significantly associated with implant survival, although sex tended to be associated with implant survival. CONCLUSIONS: Immediate-loading implant treatment for completely edentulous mandibles had acceptable clinical results in the long term. Although we could not identify significant risk factors, we established a multilevel mixed-effects parametric survival analysis with the immediate-loading implant survival data.