NaV1.1 contributes to the cell cycle of human mesenchymal stem cells by regulating AKT and CDK2.

Non-excitable cells express sodium voltage-gated channel alpha subunit 1 gene and protein (SCN1A/NaV1.1); however, the functions of NaV1.1 are unclear. SCN1A was expressed in human mesenchymal stem cells (MSCs). Nav1.1 was abundantly expressed in the endoplasmic reticulum of MSCs; however, its expression was not found to be related to sodium currents. SCN1A-silencing reduced MSC proliferation and delayed the cell cycle in the S phase. SCN1A-silencing also suppressed the protein levels of CDK2 and AKT, despite similar mRNA expression, and inhibited AKT phosphorylation in MSCs. Cycloheximide-chase assay showed that SCN1A-silencing induced CDK2 but not AKT protein degradation in MSCs. Proteolysis inhibition assay using epoxomicin, bafilomycin A1, and NH4Cl, revealed that the ubiquitin-proteasome and autophagy/endo-lysosome systems were irrelevant to CDK2 and AKT protein reduction in SCN1A-silenced MSCs. AKT inhibitor LY294002 did not affect the degradation and nuclear localization of CDK2 in MSCs. Likewise, AKT activator SC79 did not attenuate the SCN1A-silencing effects on CDK2 in MSCs. These results suggest that NaV1.1 contributes to the cell cycle of MSCs by regulating the post-translational control of AKT and CDK2.

Joint segmentation of sternocleidomastoid and skeletal muscles in computed tomography images using a multiclass learning approach.

Deep-learning-based methods can improve robustness against individual variations in computed tomography (CT) images of the sternocleidomastoid muscle, which is a challenge when using conventional methods based on probabilistic atlases are used for automatic segmentation. Thus, this study proposes a novel multiclass learning approach for the joint segmentation of the sternocleidomastoid and skeletal muscles in CT images, and it employs a two-dimensional U-Net architecture. The proposed method concurrently learns and segmented segments the sternocleidomastoid muscle and the entire skeletal musculature. Consequently, three-dimensional segmentation results are generated for both muscle groups. Experiments conducted on a dataset of 30 body CT images demonstrated segmentation accuracies of 82.94% and 92.73% for the sternocleidomastoid muscle and entire skeletal muscle compartment, respectively. These results outperformed those of conventional methods, such as the single-region learning of a target muscle and multiclass learning of specific muscle pairs. Moreover, the multiclass learning paradigm facilitated a robust segmentation performance regardless of the input image range. This highlights the method's potential for cases that present muscle atrophy or reduced muscle strength. The proposed method exhibits promising capabilities for the high-accuracy joint segmentation of the sternocleidomastoid and skeletal muscles and is effective in recognizing skeletal muscles, thus, it holds promise for integration into computer-aided diagnostic systems for comprehensive musculoskeletal analysis. These findings are expected to enhance medical image analysis techniques and their applications in clinical decision support systems.

MRI features of histological subtypes of thyroid cancer in comparison with CT findings: differentiation between anaplastic, poorly differentiated, and papillary thyroid carcinoma.

PURPOSE: This study aimed to evaluate the MRI features of the main histological subtypes of thyroid cancer and enable differentiation between anaplastic thyroid carcinoma (ATC), poorly differentiated thyroid carcinoma (PDTC), and papillary thyroid carcinoma (PTC). MATERIALS AND METHODS: This study included 79 patients with histopathologically proven thyroid cancer (14 ATCs, 8 PDTCs, and 57 PTCs) who underwent neck MRI. MRI images were retrospectively reviewed and compared between the three pathologies. RESULTS: The maximum diameter was larger in ATCs and PDTCs than in PTCs (65.2 mm and 38.4 mm vs. 26.0 mm, p < 0.01). The signal intensity ratio of the solid components on T2-weighted images (T2WIs) was higher in ATCs than in PTCs (1.13 vs. 0.89, p < 0.05). The predominant signal intensity of the solid components on T2WI exhibited hyperintensity relative to the spinal cord in ATCs more frequently than in PTCs (71% vs. 30%, p < 0.01), whereas hypointensity was more frequent in PTCs than in ATCs and PDTCs (60% vs. 0% and 13%, p < 0.01). Intratumoral ring-shaped hypointensity on T2WI was more frequent in ATCs than in PDTCs and PTCs (64% vs. 13% and 18%, p < 0.01). An ill-defined margin was more frequent in ATCs and PDTCs than in PTCs (93% and 63% vs. 25%, p < 0.01). Extrathyroidal extension, tracheal invasion, esophageal invasion, vascular invasion, and venous thrombosis were more frequently observed in ATCs than in PTCs (p < 0.05). CONCLUSIONS: MRI could characterize the differences between ATCs, PDTCs, and PTCs.

Pelvic inflammatory disease associated with cytomegalovirus infection in an immunocompetent adult: Case report and literature review.

Pelvic inflammatory disease associated with cytomegalovirus infection in immunocompetent adults might be difficult to diagnose because of the rarity and relatively inconspicuous symptoms of infectious mononucleosis. Even if the main complaint is lower abdominal pain, careful search for symptoms latent outside the abdomen could lead to the diagnosis.

Thyroid hemiatrophy associated with papillary thyroid carcinoma.

PURPOSE: The present study aimed to investigate CT imaging features, pathological findings, and prognosis in patients with thyroid hemiatrophy (THA) associated with papillary thyroid carcinoma (PTC). METHODS: This retrospective study included 225 patients with histopathologically proven PTC treated by surgical resection who underwent preoperative CT scanning. On CT images, THA was defined as thyroid parenchymal hemiatrophy on the ipsilateral side of PTC. CT findings, overall survival, and disease-free survival were compared between patients with and without THA. Pathological findings were also assessed in PTCs with and without THA. RESULTS: THA was observed in 35 of 225 (16%) patients with PTC. Atrophic thyroid parenchyma was observed in the right lobe of 20 patients (57%) and in the left lobe of the remaining 15 patients (43%). With respect to the solid components within PTCs, contrast-enhanced CT attenuation (114.2 ± 18.2 vs. 126.7 ± 31.3 HU; p < 0.05) and CT attenuation change for contrast-enhanced CT minus unenhanced CT (60.2 ± 18.1 vs. 72.3 ± 31.0 HU; p < 0.05) were significantly lower in PTCs with THA than in those without THA. Histopathologically, almost all PTCs with THA (97%) had keloid-like collagen, which is broad bundles of hypocellular collagen with bright eosinophilic hyalinization, typically observed in keloid. However, no significant differences were observed in the prognosis between the two groups. CONCLUSION: THA was occasionally observed in patients with PTC. Weak contrast-enhancement was distinct characteristic of PTC patients with THA, which is probably caused by keloid-like collagen.

Diffusion-weighted Echo Planar Imaging with Compressed SENSE (EPICS-DWI) for Pancreas Assessment: A Multicenter Study.

PURPOSE: This study aimed to evaluate the feasibility of single-shot echo planar diffusion-weighted imaging with compressed SENSE (EPICS-DWI) for pancreas assessment by comparing with single-shot echo planar DWI with parallel imaging (PI-DWI). METHODS: This multicenter prospective study included 27 consecutive participants with untreated pancreatic ductal adenocarcinoma (PDAC) (15 men; mean age, 67 ± 10 years) who underwent pancreatic protocol MRI including both PI-DWI and EPICS-DWI. Two radiologists independently and randomly reviewed the high b-value DWI images and qualitatively assigned confidence scores for overall image quality, image noise, pancreas conspicuity, and PDAC conspicuity using a 5-point scale. One radiologist measured the PDAC-to-pancreas contrast-to-noise-ratio (CNR) on high b-value DWI images and the apparent diffusion coefficient (ADC) value of PDAC. Qualitative and quantitative parameters were compared between PI-DWI and EPICS-DWI using the Wilcoxon signed-rank test. RESULTS: The confidence scores for overall image quality (P < 0.001 in both radiologists) and image noise (P < 0.001 in both radiologists) were higher in EPICS-DWI than in PI-DWI. The pancreas conspicuity was better in EPICS-DWI than in PI-DWI in one of the radiologists (P = 0.02 and 0.06). The PDAC conspicuity was comparable between PI-DWI and EPICS-DWI (P > 0.99 in both radiologists). The PDAC-to-pancreas CNR was higher in EPICS-DWI than in PI-DWI (P = 0.02), while the ADC value of PDAC in PI-DWI was not significantly different compared to that in EPICS-DWI (P = 0.48). CONCLUSION: The image quality and PDAC-to-pancreas CNR was improved in EPICS-DWI compared to PI-DWI. However, the conspicuity and ADC value of PDAC were comparable between PI-DWI and EPICS-DWI.

Does Colorectal Stenting as a Bridge to Surgery for Obstructive Colorectal Cancer Increase Perineural Invasion?

Objectives: To clarify whether self-expandable metallic stent (SEMS) placement for obstructive colorectal cancer (CRC) increases perineural invasion (PNI), thereby worsening the prognosis. Methods: In total, 1022 patients with pathological T3 or T4 colon or rectosigmoid cancer who underwent resection were retrospectively reviewed. The study patients were divided into a no obstruction group (n=693), obstruction without stent group (n=251), and obstruction with stent group (n=78), and factors demonstrating an independent association with PNI, the difference in PNI incidence and severity between groups, and the association between PNI and the duration from SEMS placement to surgery were investigated. Survival analysis was performed for each group. Results: On multivariate analysis, SEMS placement (hazard ratio [HR]: 2.08) was independently associated with PNI whereas SEMS placement was not. PNI occurred in 39%, 45%, and 68% of the no obstruction, obstruction without stent, and obstruction with stent group, respectively. In the obstruction with stent group, the proportion of PNI was not associated with the duration from SEMS placement to surgery. Extramural PNI, an advanced form of PNI, demonstrated no increase with increasing interval. The five-year OS was 86.3%, 76.7%, and 73.1% in no obstruction, obstruction without stent, and obstruction with stent group, respectively. On multivariate analysis, obstruction was an independent risk factor of decreased OS (HR: 1.57) whereas SEMS placement was not. Conclusions: The prognosis was comparable between patients with SEMS placement and those with an obstruction who did not undergo SEMS placement, thus demonstrating that SEMS is a viable, therapeutic option for BTS.

Daptomycin treatment for persistent bacteremia in a pediatric solid organ transplant recipient: Case report and literature review.

INTRODUCTION: Information on the clinical utility of daptomycin in patients with persistent bacteremia and daptomycin's pharmacokinetic data in pediatric patients has been sparse. In addition, reports on the experience of using daptomycin in children undergoing solid organ transplantation have been extremely limited. The authors describe a pediatric case of persistent bacteremia after solid organ transplantation successfully treated by daptomycin. Blood daptomycin concentrations were measured by liquid chromatography-mass spectrometry and pharmacokinetic analysis was performed. We also conducted a literature review on the use of daptomycin in children with persistent bacteremia. CASE REPORT: An eight-year-old girl who underwent small bowel and liver transplantation experienced persistent bacteremia due to Staphylococcus epidermidis. The bacteremia persisted despite standard therapy; however, it finally resolved with the addition of daptomycin. The patient had renal dysfunction and the initial dosing resulted in excessive drug exposure. The dosage was adjusted based on the pharmacokinetic analysis. The dosage of administrated teicoplanin was also adjusted according to trough concentration values. In the literature review, we identified 12 cases of neonates and 24 cases of post-neonatal children with the experience of using daptomycin for persistent bacteremia; however, no solid organ transplant recipient was identified. Similar trends in blood concentrations and dose ratios of teicoplanin and daptomycin were observed over time. DISCUSSION: More information is required regarding the clinical utility and pharmacokinetics of daptomycin in pediatric patients with persistent bacteremia. Referring to the exposure to renally excreted drugs that are routinely measured and pharmacokinetic analysis of daptomycin may be useful in optimizing the dose of daptomycin in special patient populations, including those with renal impairment.

An N-terminal and ankyrin repeat domain interactome of Shank3 identifies the protein complex with the splicing regulator Nono in mice.

An autism-associated gene Shank3 encodes multiple splicing isoforms, Shank3a-f. We have recently reported that Shank3a/b-knockout mice were more susceptible to kainic acid-induced seizures than wild-type mice at 4 weeks of age. Little is known, however, about how the N-terminal and ankyrin repeat domains (NT-Ank) of Shank3a/b regulate multiple molecular signals in the developing brain. To explore the functional roles of Shank3a/b, we performed a mass spectrometry-based proteomic search for proteins interacting with GFP-tagged NT-Ank. In this study, NT-Ank was predicted to form a variety of complexes with a total of 348 proteins, in which RNA-binding (n = 102), spliceosome (n = 22), and ribosome-associated molecules (n = 9) were significantly enriched. Among them, an X-linked intellectual disability-associated protein, Nono, was identified as a NT-Ank-binding protein. Coimmunoprecipitation assays validated the interaction of Shank3 with Nono in the mouse brain. In agreement with these data, the thalamus of Shank3a/b-knockout mice aberrantly expressed splicing isoforms of autism-associated genes, Nrxn1 and Eif4G1, before and after seizures with kainic acid treatment. These data indicate that Shank3 interacts with multiple RNA-binding proteins in the postnatal brain, thereby regulating the homeostatic expression of splicing isoforms for autism-associated genes after birth.

A study of traction endoscopic submucosal dissection using a unique traction device with a ring and a clip with an obtuse tip.

Appropriate use of traction devices is important in Endoscopic Submucosal Dissection (ESD). The Sure-Clip Traction Band (Microtec: "traction band") is a disposable clip with a series of three small silicone rings, which can be used as a traction device by inserting the clip through two holes. The SB clip long with a 125-degree tip claw angle (SB Kawasumi: hereafter referred to as SB clip) has an obtuse angle tip, can be re-gripped and is considered to cause less damage to the digestive tract wall when withdrawn after clipping. We performed traction ESD using a traction band and SB clip. These were used for gastric ESD in 4 cases and colorectal ESD in 3 cases.

Experience with the new multi-hole self-expandable metallic stent.

Multi-hole self-expandable metallic stent (M.I.Tech/Boston Scientific: MHSEMS) has small holes in the cover part of FCSEMS and has properties between UCSEMS and FCSEMS. The MHSEMS was originally an FCSEMS with holes in all cells. In the distal bile duct, the MHSEMS have the potential to prevent migration by allowing tissue to enter the small holes. In the hilar region, the MHSEMS can potentially prolong the duration of patency of the central bile duct by preventing ingrowth by covering the branches without impeding them. However, because there were reports of cases of inoperable removal due to ingrowth overseas, the model launched in Japan in 2022 had fewer holes, and the cover of the Hanarostent Biliary Full Cover NEO (M.I. Tech/Boston Scientific) had a row of holes in every other row. It was used in 7 cases of malignant biliary obstruction. The primary diseases were gallbladder cancer, cholangiocarcinoma, and pancreatic cancer. The obstruction sites were the hilar region and the distal bile duct. All patients underwent successful implantation with no pancreatitis or cholangitis observed. Five patients continued chemotherapy. One gallbladder cancer patient died after 20 days, one cholangiocarcinoma patient experienced obstruction after 81 days, and the other five cases showed no evidence of obstruction or deviation (ranging from 6 to 119 days after implantation, with an average of 75 days). MHSEMS can be used for malignant biliary obstruction, mainly in the distal bile duct.

Gnao1 is a molecular switch that regulates the Rho signaling pathway in differentiating neurons.

GNAO1 encodes G protein subunit alpha O1 (Gαo). Pathogenic variations in GNAO1 cause developmental delay, intractable seizures, and progressive involuntary movements from early infancy. Because the functional role of GNAO1 in the developing brain remains unclear, therapeutic strategies are still unestablished for patients presenting with GNAO1-associated encephalopathy. We herein report that siRNA-mediated depletion of Gnao1 perturbs the expression of transcripts associated with Rho GTPase signaling in Neuro2a cells. Consistently, siRNA treatment hampered neurite outgrowth and extension. Growth cone formation was markedly disrupted in monolayer neurons differentiated from iPSCs from a patient with a pathogenic variant of Gαo (p.G203R). This variant disabled neuro-spherical assembly, acquisition of the organized structure, and polarized signals of phospho-MLC2 in cortical organoids from the patient's iPSCs. We confirmed that the Rho kinase inhibitor Y27632 restored these morphological phenotypes. Thus, Gαo determines the self-organizing process of the developing brain by regulating the Rho-associated pathway. These data suggest that Rho GTPase pathway might be an alternative target of therapy for patients with GNAO1-associated encephalopathy.

Assessment of Arterial Involvement in Pancreatic Cancer: Utility of Reconstructed CT Images Perpendicular to Artery.

The purpose of this study was to investigate the utility of reconstructed CT images perpendicular to the artery for assessing arterial involvement from pancreatic cancer and compare the interobserver variability between it and the current diagnostic imaging method. This retrospective study included patients with pancreatic cancer in the pancreatic body or tail who underwent preoperative pancreatic protocol CT and distal pancreatectomy. Five radiologists used axial and coronal CT images (current method) and perpendicular reconstructed CT images (proposed method) to determine if the degree of solid soft-tissue contact with the splenic artery was ≤180° or >180°. The generalized estimating equations were used to compare the diagnostic performance of solid soft-tissue contact >180° between the current and proposed methods. Fleiss' ĸ statistics were used to assess interobserver variability. The sensitivity and negative predictive value for diagnosing solid soft-tissue contact >180° were higher (p < 0.001 for each) and the specificity (p = 0.003) and positive predictive value (p = 0.003) were lower in the proposed method than the current method. Interobserver variability was improved in the proposed method compared with the current method (ĸ = 0.87 vs. 0.67). Reconstructed CT images perpendicular to the artery showed higher sensitivity and negative predictive value for diagnosing solid soft-tissue contact >180° than the current method and demonstrated improved interobserver variability.

Soluble form of the MDA5 protein in human sera.

Viral double-stranded RNA (dsRNA) is sensed by toll-like receptor 3 (TLR3) and retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs), including melanoma differentiation-associated gene 5 (MDA5). MDA5 recognizes the genome of dsRNA viruses and replication intermediates of single-stranded RNA viruses. MDA5 also plays an important role in the development of autoimmune diseases, such as Aicardi-Goutieres syndrome and type I diabetes. Patients with dermatomyositis with serum MDA5 autoantibodies (anti-CADM-140) are known to have a high risk of developing rapidly progressive interstitial lung disease and poor prognosis. However, there have been no reports on the soluble form of MDA5 in human serum. In the present study, we generated in-house monoclonal antibodies (mAbs) against human MDA5. We then performed immunohistochemical analysis and sensitive sandwich immunoassays to detect the MDA5 protein using two different mAbs (clones H27 and H46). As per the immunohistochemical analysis, the MDA5 protein was moderately expressed in the alveolar epithelia of normal lungs and was strongly expressed in the cytoplasm of lymphoid cells in the tonsils and acinar cells of the pancreas. Interestingly, soluble MDA5 protein was detectable in the serum, but not in the urine, of healthy donors. Soluble MDA5 protein was also detectable in the serum of patients with dermatomyositis. Immunoblot analysis showed that human cells expressed a 120 kDa MDA5 protein, while the 60 kDa MDA5 protein increased in the supernatant of peripheral mononuclear cells within 15 min after MDA5 agonist/double-strand RNA stimulation. Hydrogen deuterium exchange mass spectrometry revealed that an anti-MDA5 mAb (clone H46) bound to the epitope (415QILENSLLNL424) derived from the helicase domain of MDA5. These results indicate that a soluble MDA5 protein containing the helicase domain of MDA5 could be rapidly released from the cytoplasm of tissues after RNA stimulation.

Recurrence of rectal cancer on the pelvic sidewall after lateral lymph node dissection.

PURPOSE: Although lateral lymph node dissection has been performed to prevent lateral pelvic recurrence in locally advanced lower rectal cancer, the incidence of lateral pelvic recurrence after this procedure has not been investigated. Therefore, this study aimed to investigate the long-term outcomes of patients who underwent lateral pelvic lymph node dissection, with a particular focus on recurrence patterns. METHODS: This was a retrospective study conducted at a single high-volume cancer center in Japan. A total of 493 consecutive patients with stage II-III rectal cancer who underwent lateral lymph node dissection between January 2005 and August 2022 were included. The primary outcome measures included patterns of recurrence, overall survival, and relapse-free survival. Patterns of recurrence were categorized as lateral or central pelvic. RESULTS: Among patients who underwent lateral lymph node dissection, 18.1% had pathologically positive lateral lymph node metastasis. Lateral pelvic recurrence occurred in 5.5% of patients after surgery. Multivariate analysis identified age > 75 years, lateral lymph node metastasis, and adjuvant chemotherapy as independent risk factors for lateral pelvic recurrence. Evaluation of the recurrence rate by dissection area revealed approximately 1% of recurrences in each area after dissection. CONCLUSION: We demonstrated the prognostic outcome and limitations of lateral lymph node dissection for patients with advanced lower rectal cancer, focusing on the incidence of recurrence in the lateral area after the dissection. Our study emphasizes the clinical importance of lateral lymph node dissection, which is an essential technique that surgeons should acquire.

ATP1A3 regulates protein synthesis for mitochondrial stability under heat stress.

Pathogenic variants in ATP1A3, the gene encoding the α3 subunit of the Na+/K+-ATPase, cause alternating hemiplegia of childhood (AHC) and related disorders. Impairments in Na+/K+-ATPase activity are associated with the clinical phenotype. However, it remains unclear whether additional mechanisms are involved in the exaggerated symptoms under stressed conditions in patients with AHC. We herein report that the intracellular loop (ICL) of ATP1A3 interacted with RNA-binding proteins, such as Eif4g (encoded by Eif4g1), Pabpc1 and Fmrp (encoded by Fmr1), in mouse Neuro2a cells. Both the siRNA-mediated depletion of Atp1a3 and ectopic expression of the p.R756C variant of human ATP1A3-ICL in Neuro2a cells resulted in excessive phosphorylation of ribosomal protein S6 (encoded by Rps6) and increased susceptibility to heat stress. In agreement with these findings, induced pluripotent stem cells (iPSCs) from a patient with the p.R756C variant were more vulnerable to heat stress than control iPSCs. Neurons established from the patient-derived iPSCs showed lower calcium influxes in responses to stimulation with ATP than those in control iPSCs. These data indicate that inefficient protein synthesis contributes to the progressive and deteriorating phenotypes in patients with the p.R756C variant among a variety of ATP1A3-related disorders.