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Our civilization creates enormous volumes of digital data, a substantial fraction of which is preserved and made publicly available for present and future usage. Additionally, historical born-analog records are progressively being digitized and incorporated into digital document repositories. While professionals often have a clear idea of what they are looking for in document archives, average users are likely to have no precise search needs when accessing available archives (e.g., through their online interfaces). Thus, if the results are to be relevant and appealing to average people, they should include engaging and recognizable material. However, state-of-the-art document archival retrieval systems essentially use the same approaches as search engines for synchronic document collections. In this article, we develop unique ranking criteria for assessing the usefulness of archived contents based on their estimated relationship with current times, which we call contemporary relevance. Contemporary relevance may be utilized to enhance access to archival document collections, increasing the likelihood that users will discover interesting or valuable material. We next present an effective strategy for estimating contemporary relevance degrees of news articles by utilizing learning to rank approach based on a variety of diverse features, and we then successfully test it on the New York Times news collection. The incorporation of the contemporary relevance computation into archival retrieval systems should enable a new search style in which search results are meant to relate to the context of searchers' times, and by this have the potential to engage the archive users. As a proof of concept, we develop and demonstrate a working prototype of a simplified ranking model that operates on the top of the Portuguese Web Archive portal (arquivo.pt). [ABSTRACT FROM AUTHOR]

Aim: Dysphagia is a problem typically associated with aging. The aim was to investigate the relationship between dysphagia and motor function using a simple assessment method that can be performed in the community setting, and to promote the early detection and prevention of dysphagia. Methods: Data from the Locomotive Syndrome and Health Outcome in Aizu Cohort Study (LOHAS) were used. Those aged ≥65 years were included. Motor function was assessed using a grip strength test, single limb standing test (SLS), and timed up and go test (TUG). Swallowing function was assessed using the Japanese version of the 10‐item Eating Assessment Tool (EAT‐10). The association between motor function and swallowing function was analyzed. Results: In total, 1732 participants were included. In logistic regression modes in which grip strength, SLS, and TUG results were included separately, the odds ratio for dysphagia increased by 1.08 (P = 0.001) for each 1‐kg decrease in grip strength, and increased by 1.15 (P < 0.001) for each 1‐s increase in TUG time. No association was found for SLS. In the model in which grip strength and TUG were included simultaneously, the odds ratio for dysphagia increased by 1.06 (P = 0.01) in grip strength, and increased by 1.11 (P = 0.009) in TUG time. Conclusion: Our results suggest that skeletal muscle strength and dynamic balance function are associated with dysphagia in community‐dwelling older people. Geriatr Gerontol Int 2023; 23: 603–608. [ABSTRACT FROM AUTHOR]

Recent years have witnessed an evolution of imaging technologies towards sophisticated approaches for visualising cells within their natural environment(s) and for investigating their interactions with other cells, with adjacent anatomical structures, and with implanted biomaterials. Resin cast etching (RCE) is an uncomplicated technique involving sequential acid etching and alkali digestion of resin embedded bone to observe the osteocyte lacuno-canalicular network using scanning electron microscopy. This review summarises the applicability of RCE to bone and the bone-implant interface. Quantitative parameters such as osteocyte size, osteocyte density, and number of canaliculi per osteocyte, and qualitative metrics including osteocyte shape, disturbances in the arrangement of osteocytes and canaliculi, and physical communication between osteocytes and implant surfaces can be investigated. Ageing, osteoporosis, long-term immobilisation, spinal cord injury, osteoarthritis, irradiation, and chronic kidney disease have been shown to impact osteocyte lacuno-canalicular network morphology. In addition to titanium, calcium phosphates, and bioactive glass, observation of direct connectivity between osteocytes and cobalt chromium provides new insights into the osseointegration potential of materials conventionally viewed as non-osseointegrating. Other applications include in vivo and in vitro testing of polymer-based tissue engineering scaffolds and tissue-engineered ossicles, validation of ectopic osteochondral defect models, ex vivo organ culture of whole bones, and observing the effects of gene dysfunction/deletion on the osteocyte lacuno-canalicular network. Without additional contrast staining, any resin embedded specimen (including clinical biopsies) can be used for RCE. The multitude of applications described here attest to the versatility of RCE for routine use within correlative analytical workflows, particularly in biomaterials science. [ABSTRACT FROM AUTHOR]

Control of angiogenesis is widely considered a therapeutic strategy, but reliable control methods are still under development. Phosphorylation of myosin light chain 2 (MLC2), which regulates actin-myosin interaction, is critical to the behavior of vascular endothelial cells (ECs) during angiogenesis. MLC2 is phosphorylated by MLC kinase (MLCK) and dephosphorylated by MLC phosphatase (MLCP) containing a catalytic subunit PP1. We investigated the potential role of MLC2 in the pharmacological control of angiogenesis. We exposed transgenic zebrafish Tg(fli1a:Myr-mCherry) ncv1 embryos to chemical inhibitors and observed vascular development. PP1 inhibition by tautomycetin increased length of intersegmental vessels (ISVs), whereas MLCK inhibition by ML7 decreased it; these effects were not accompanied by structural dysplasia. ROCK inhibition by Y-27632 also decreased vessel length. An in vitro angiogenesis model of human umbilical vein endothelial cells (HUVECs) showed that tautomycetin increased vascular cord formation, whereas ML7 and Y-27632 decreased it. These effects appear to be influenced by regulation of cell morphology rather than cell viability or motility. Actin co-localized with phosphorylated MLC2 (pMLC2) was abundant in vascular-like elongated-shaped ECs, but poor in non-elongated ECs. pMLC2 was associated with tightly arranged actin, but not with loosely arranged actin. Moreover, knockdown of MYL9 gene encoding MLC2 reduced total MLC2 and pMLC2 protein and inhibited angiogenesis in HUVECs. The present study found that MLC2 is a pivotal regulator of angiogenesis. MLC2 phosphorylation may be involved in the regulation of of cell morphogenesis and cell elongation. The functionally opposite inhibitors positively or negatively control angiogenesis, probably through the regulating EC morphology. These findings may provide a unique therapeutic target for angiogenesis. • PP1 inhibitors show angiogenic effects in zebrafish embryos and human ECs. • MLCK inhibitors show anti- angiogenic effects in zebrafish embryos and ECs. • PP1 and MLCK inhibitors affect EC elongation, but not cell viability or migration. • Elongated-shaped ECs show co-localization of pMLC2 and F-actin. • Knockdown of the MYL9 gene encoding MLC2 inhibits EC migration and morphological activity and suppresses angiogenesis. [ABSTRACT FROM AUTHOR]

Staphylococcus epidermidis are gram-positive bacteria that form a biofilm around implanted devices and develop an infection into a chronic state. Recently, it has been revealed that microvesicles have important roles in biofilm formation and intercellular communication among bacteria. However, biofilm formation of Staphylococcus epidermidis, and its relation to microvesicle secretion, is poorly understood because of the difficulty required to preserve the delicate water-rich morphology of biofilm for high-resolution observations. Here, we successfully imaged the microvesicles secreted from Staphylococcus epidermidis and the subsequent process of their integration into biofilm using liquid–phase imaging using atmospheric scanning electron microscopy (ASEM). In the biofilm, cells were connected by nanotube-like structures attached by microvesicles, and surrounded by extracellular polymeric substances. Cells cultured in the ASEM specimen holder were aldehyde-fixed and stained using positively charged nanogold labelling and/or using National Center for Microscopy and Imaging Research method. The samples immersed in aqueous radical scavenger glucose buffer were imaged by the inverted SEM of ASEM. Information regarding the morphologies of microvesicles, nanotube-like fibrils, and biofilm formed by Staphylococcus epidermidis is expected to be useful to elucidate the biological mechanism of biofilm formation and to develop a medicine against biofilms and their associated infections. [ABSTRACT FROM AUTHOR]

Chondrogenesis and angiogenesis drive endochondral ossification. Using the atmospheric scanning electron microscopy (ASEM) without decalcification and dehydration, we directly imaged angiogenesis-driven ossification at different developmental stages shortly after aldehyde fixation, using aqueous radical scavenger glucose solution to preserve water-rich structures. An embryonic day 15.5 mouse femur was fixed and stained with phosphotungstic acid (PTA), and blood vessel penetration into the hypertrophic chondrocyte zone was visualised. We observed a novel envelope between the perichondrium and proliferating chondrocytes, which was lined with spindle-shaped cells that could be borderline chondrocytes. At postnatal day (P)1, trabecular and cortical bone mineralisation was imaged without staining. Additional PTA staining visualised surrounding soft tissues; filamentous connections between osteoblast-like cells and osteocytes in cortical bone were interpreted as the osteocytic lacunar-canalicular system. By P10, resorption pits had formed on the tibial trabecular bone surface. The applicability of ASEM for pathological analysis was addressed using knockout mice of Keap1, an oxidative-stress sensor. In Keap1−/− femurs, we observed impaired calcification and angiogenesis of epiphyseal cartilage, suggesting impaired bone development. Overall, the quick ASEM method we developed revealed mineralisation and new structures in wet bone tissue at EM resolution and can be used to study mineralisation-associated phenomena of any hydrated tissue. [ABSTRACT FROM AUTHOR]

Glycine, a non-essential amino acid, exerts concentration-dependent biphasic effects on angiogenesis. Low-doses of glycine promote angiogenesis, whereas high-doses cause anti-angiogenesis. The phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling participates in angiogenesis of both physiological development, and pathological events including tumor and inflammation. We assessed the role of PI3K/Akt/mTOR signaling in vascular development, and the interaction with glycine, using transgenic zebrafish Tg(fli1a:Myr-mCherry) ncv1 embryos expressing fluorescent proteins in vascular endothelial cells. Treatment with inhibitors of mTORC1 (rapamycin and everolimus), mTORC1/mTORC2 (KU0063794), PI3K (LY29400), and Akt (Akt inhibitor) decreased the development of intersegmental vessels (ISVs). These inhibitors cancelled the angiogenic effects of a low-dose of glycine, while acted synergistically with a high-dose of glycine in anti-angiogenesis. mTOR signaling regulates the gene expression of vascular endothelial growth factor (VEGF) , a major angiogenic factor, and nitric oxide (NO) synthase (NOS) , an enzyme for the synthesis of an angiogenic mediator NO. Expressions of VEGF and NOS were consistent with the vascular features induced by glycine and an mTOR inhibitor. Our results suggest that PI3K/Akt/mTOR signaling may interact with dose-dependent biphasic effects of exogenous glycine on in vivo angiogenesis. mTOR signaling is a key target for cancer therapy, thus, the combining mTOR inhibitors with glycine may be a potential approach for controlling angiogenesis. • Inhibition of PI3K/Akt/mTOR signaling impairs vascular development. • Low-glycine induced angiogenesis is blocked by inhibition of mTOR signaling. • High-glycine induced anti-angiogenesis is enhanced by inhibition of mTOR signaling. • Combination of glycine and an mTOR inhibitor affects expression of VEGF and NOS genes. [ABSTRACT FROM AUTHOR]

Glycine, a non-essential amino acid, is involved in both angiogenesis and anti-angiogenesis. We hypothesized that glycine would exert dose-dependent different effects on angiogenesis. In this study, we investigated the effects of a broad range of concentrations of glycine on vascular development using transgenic zebrafish Tg(fli1a:Myr-mCherry) ncv1 embryos. Effects of glycine transporter (GlyT) inhibitors (sarcosine and bitopertin) and a glycine receptor (GlyR) inhibitor (strychnine) were also examined in embryos in the absence or presence of glycine. After exposure to glycine and inhibitors, intersegmental vessels (ISVs) were observed by fluorescent microscopy. Low concentrations of glycine promoted the development of ISVs, whereas high concentrations reduced it. These effects of glycine could generally be reversed by treatment with GlyT and GlyR inhibitors. Furthermore, expressions of vascular endothelial growth factor (VEGF) (an angiogenic factor) and nitric oxide synthase (NOS) (an enzyme for nitric oxide synthesis) were associated with the dose-dependent effects of glycine. Our results suggest that glycine exerts dose-dependent biphasic effects on vascular development, which rely on GlyTs and GlyRs, and correlate with the expression of VEGF and NOS genes. At low concentrations, glycine acted as an angiogenic factor. In contrast, at high concentrations, glycine induced anti-angiogenesis. This evidence provides a novel insight into glycine as a unique target in angiogenic and anti-angiogenic therapy. • Glycine exerts biphasic effects on the vascular development of zebrafish embryos. • Inhibition of glycine transporters and receptors impairs vascular development. • Effects of glycine are blocked by inhibition of glycine transporters and receptors. • Glycine affects the expression of VEGF and NOS genes. [ABSTRACT FROM AUTHOR]

Opsin3 (Opn3) is a transmembrane heptahelical G protein–coupled receptor (GPCR) with the potential to produce a nonvisual photoreceptive effect. Interestingly, anatomical profiling of GPCRs reveals that Opn3 mRNA is highly expressed in adipose tissue. The photosensitive functions of Opn3 in mammals are poorly understood, and whether Opn3 has a role in fat is entirely unknown. In this study, we found that Opn3-knockout (Opn3-KO) mice were prone to diet-induced obesity and insulin resistance. At the cellular level, Opn3-KO brown adipocytes cultured in darkness had decreased glucose uptake and lower nutrient-induced mitochondrial respiration than wild-type (WT) cells. Light exposure promoted mitochondrial activity and glucose uptake in WT adipocytes but not in Opn3-KO cells. Brown adipocytes carrying a defective mutation in Opn3's putative G protein–binding domain also exhibited a reduction in glucose uptake and mitochondrial respiration in darkness. Using RNA-sequencing, we identified several novel light-sensitive and Opn3-dependent molecular signatures in brown adipocytes. Importantly, direct exposure of brown adipose tissue (BAT) to light in living mice significantly enhanced thermogenic capacity of BAT, and this effect was diminished in Opn3-KO animals. These results uncover a previously unrecognized cell-autonomous, light-sensing mechanism in brown adipocytes via Opn3-GPCR signaling that can regulate fuel metabolism and mitochondrial respiration. Our work also provides a molecular basis for developing light-based treatments for obesity and its related metabolic disorders. Brown adipose tissue plays a pivotal role in energy homeostasis and serves as a metabolic sink for glucose and fatty acid. This study demonstrates a novel light-sensing mechanism in mice via the photoreceptor Opsin3 that regulates fuel utilization and mitochondrial activity of brown adipocytes. [ABSTRACT FROM AUTHOR]

We examined the experiences of a public health nurse (PHN) in a rural area affected by the Great East Japan Earthquake. We used an ethnographic method to obtain in-depth information about her experiences, from which six themes were developed. The PHN risked her life to protect members of the community, but she remained anxious to see her own young children as soon as possible. Nevertheless, she was strongly committed to helping community members and continued to work for them. We suggest creating a practical system to allow PHNs to obtain information about the safety of their own families during a disaster. [ABSTRACT FROM PUBLISHER]

Highlights: [•] BMP2 induces Y1 receptor expression in C2C12 cells. [•] Smad1/4 induces Y1 receptor expression in C2C12 cells. [•] Y1 gene promoter activity is regulated by BMP2 signaling. [•] Y1 receptor suppression by siRNA increases osteoblast-related gene expression. [ABSTRACT FROM AUTHOR]

The adhesion of leukocytes circulating in the blood to vascular endothelium is critical for their trafficking in the vasculature, and CD44 is an important cell surface receptor for rolling adhesion. In this study, we demonstrate the correlative observation of CD44 distribution at the lymphocyte cell surface in liquid by fluorescence optical microscopy and immuno-electron microscopy using an atmospheric scanning electron microscope (ASEM). The ultrastructure of the cell surface was clearly imaged by ASEM using positively charged Nanogold particles. ASEM analysis demonstrated microvilli projections around the cell surface and the localization of CD44 on the microvilli. Treatment of cells with cytochalasin D resulted in a loss of the microvilli projections and concomitantly abrogated CD44-mediated adhesion to its ligand hyaluronan. These results suggest the functional relevance of microvilli in CD44-mediated rolling adhesion under shear flow. [ABSTRACT FROM AUTHOR]

In eukaryotic cells, degradation of most intracellular proteins is carried out by the ubiquitin-proteasome pathway. Recent investigations suggest that bone metabolism is also regulated by this pathway. The clinical efficacy of bortezomib, a 26 S proteasome inhibitor used as an anticancer drug, has been linked to an increase in bone formation. In this study, we show that proteasome inhibitors induce expression of osteoblastic differentiation-related genes such as osteocalcin and alkaline phosphatase in C2 C12 cells. In contrast, myogenic differentiation is inhibited. Among the proteasome inhibitors tested, bortezomib induced the greatest increase in osteocalcin expression. Although these effects were similar to that of bone morphogenetic protein ( BMP) 2, proteasome inhibitors did not induce transcriptional activity of Smad1/4-dependent reporter or BMP2 signaling target gene expression. Transient transfection of osteocalcin promoter-luciferase constructs with bortezomib resulted in an increase in luciferase activity. Mutation of OSE2, but not OSE1, sites of the osteocalcin promoter diminished the bortezomib-induced activity. Also, Runx2 binding activity and protein levels were induced by bortezomib treatment. These results suggest that the bortezomib induces osteoblastic differentiation by modifying the activity of Runx2 and that the function of the proteasome in controlling degradation of differentiation-related transcription factors plays an important role in osteoblast differentiation. [ABSTRACT FROM AUTHOR]

Abstract: CXCL12 (stromal cell-derived factor-1, SDF-1), produced by stromal and endothelial cells including cells of the bone marrow, binds to its receptor CXCR4 and this axis regulates hematopoietic cell trafficking. Recently, osteoclast precursor cells were found to express CXCR4 and a potential role for the CXCL12–CXCR4 axis during osteoclast precursor cell recruitment/retention and development was proposed as a regulator of bone resorption. We examined the role of canonical Wnt signaling in regulating the expression of CXCL12 in bone marrow stromal cells. In mouse stromal ST2 cells, CXCL12 mRNA was expressed, while its expression was reduced in Wnt3a over-expressing ST2 (Wnt3a-ST2) cells or by treatment with lithium chloride (LiCl). Wnt3a decreased CXCL12 levels in culture supernatants from mouse bone marrow stromal cells. The culture supernatant from Wnt3a-ST2 cells also reduced migratory activity of bone marrow-derived cells in a Transwell migration assay. Silencing of glycogen synthase kinase-3β decreased CXCL12 expression, suggesting that the canonical Wnt signaling pathway regulates CXCL12 expression. In a transfection assay, LiCl down-regulated the activity of a reporter gene, a 1.8kb fragment of the 5′-flanking region of the CXCL12 gene. These results show that canonical Wnt signaling regulates CXCL12 gene expression at the transcriptional level, and this is the first study linking chemokine expression to canonical Wnt signaling. [Copyright &y& Elsevier]

Abstract: MicroRNAs (miRNAs) are small non-coding RNAs that are emerging as important post-transcriptional gene regulators. miR-206 is unique in that it is expressed only in skeletal muscle, including the myoblastic C2C12 cell line. In C2C12 cells, miR-206 expression was reduced dramatically after bone morphogenetic protein (BMP)-2 treatment. The down-regulation of miR-206 expression was also observed after co-transfection with constitutively-active Smad1 and Smad4, which are the intracellular signaling molecules of the BMP pathway. BMP-2 also reduced miR-206 expression in the presence of α-amanitin in a similar manner to that in the absence of α-amanitin. Moreover, the expression of pri-miR-206 was increased upon BMP-2 treatment for 6h compared to that in the absence of BMP-2. These results suggested that BMP-2 down-regulates miR-206 expression at the post-transcriptional level, by inhibiting the processing of pri-miR-206 into mature miR-206, and that BMP-2 could regulate miRNA biogenesis by a novel mechanism. [Copyright &y& Elsevier]

Wnt/β-catenin signaling plays an important role in the developing skeletal system. Our previous studies demonstrated that Wnt/β-catenin signaling inhibits the ability of bone morphogenetic protein (BMP)-2 to suppress myotube formation in the multipotent mesenchymal cell line C2C12 and that this inhibition is mediated by Id1. In this study, we examined the role of intracellular signaling by Wnt/β-catenin and BMP-2 in regulating the expression of osteoprotegerin (OPG) and of the receptor activator of NFκB ligand (RANKL). OPG expression was induced by Wnt/β-catenin signaling in C2C12 cells and osteoblastic MC3T3-E1 cells. Silencing of glycogen synthase kinase-3β also increased OPG expression. In contrast, R expression was suppressed by Wnt/β-catenin signaling. In a transfection assay, β-catenin induced the activity of a reporter gene, a 1.5 kb fragment of the 5′-flanking region of the OPG gene. Deletion and mutation analysis revealed that Wnt/β-catenin signaling regulates transcription of OPG via a promoter region containing two Wnt/β-catenin responsive sites. BMP-2 enhanced Wnt/β-catenin-dependent transcriptional activation of the OPG promoter. In response to BMP-2 stimulation, Smad 1 and 4 interacted with Wnt/β-catenin responsive sites. These results show that the regulation of OPG expression is mediated through two transcription pathways that involve the OPG promoter. [ABSTRACT FROM AUTHOR]

A 69-year-old woman visited our outpatient clinic due to abdominal pain and anorexia. She received initial diagnostic workup, and was admitted to the hospital as an inpatient because of ileus. CT showed tumor with a diameter of 3cm in upper left abdomen, liver metastasis, peritoneal fluid localized in the pelvic cavity, and peritoneal node. Upper and lower gastrointestinal endoscopy showed no abnormalities, and percutaneous liver biopsy confirmed adenocarcinoma. PET showed a focal abnormal uptake in proximal jejunum, but no abnormalities in pancreas and biliary tract. We diagnosed the patient with small bowel cancer. It was considered unresectable, and FOLFOX chemotherapy was initiated. However, during FOLFOX chemotherapy after 1st course, CT showed that exacerbation of primary lesion and liver mass, and significant increase in ascites. She subsequently died of pneumonia day 32 after the start of chemotherapy. Autopsy findings revealed primary small bowel adenocarcinoma with proximal jejunum. Histological response was Grade1a. The incidence of small bowel adenocarcinoma (SBA) is very low, accounting for 0.1%-0.3% of all gastrointestinal malignant neoplasms. Many cases have been reported to be within 50 cm of Treitz ligament or Bauhin valve. For diagnosis of SBA, some reports suggest small bowel series, gastrointestinal endoscope, CT, and PET are useful. However, in many advanced cases it is difficult to apply curative therapy at the time of discovery. There is no standard treatment especially for unresectable small bowel cancer, and the prognosis is poor. In this case SBA was confirmed by metastatic lesion biopsy and exclusion diagnosis. Because the disease was already advanced at the time of diagnosis and furthermore time was necessary for diagnostic workup, it caused deterioration of the general condition. As a result, adequate chemotherapy could not be performed. There is a need for further case studies to establish diagnosis and treatment of SBA. [ABSTRACT FROM AUTHOR]

A 69-year-old woman visited our outpatient clinic due to abdominal pain and anorexia. She received initial diagnostic workup, and was admitted to the hospital as an inpatient because of ileus. CT showed tumor with a diameter of 3cm in upper left abdomen, liver metastasis, peritoneal fluid localized in the pelvic cavity, and peritoneal node. Upper and lower gastrointestinal endoscopy showed no abnormalities, and percutaneous liver biopsy confirmed adenocarcinoma. PET showed a focal abnormal uptake in proximal jejunum, but no abnormalities in pancreas and biliary tract. We diagnosed the patient with small bowel cancer. It was considered unresectable, and FOLFOX chemotherapy was initiated. However, during FOLFOX chemotherapy after 1st course, CT showed that exacerbation of primary lesion and liver mass, and significant increase in ascites. She subsequently died of pneumonia day 32 after the start of chemotherapy. Autopsy findings revealed primary small bowel adenocarcinoma with proximal jejunum. Histological response was Grade1a. The incidence of small bowel adenocarcinoma (SBA) is very low, accounting for 0.1%-0.3% of all gastrointestinal malignant neoplasms. Many cases have been reported to be within 50 cm of Treitz ligament or Bauhin valve. For diagnosis of SBA, some reports suggest small bowel series, gastrointestinal endoscope, CT, and PET are useful. However, in many advanced cases it is difficult to apply curative therapy at the time of discovery. There is no standard treatment especially for unresectable small bowel cancer, and the prognosis is poor. In this case SBA was confirmed by metastatic lesion biopsy and exclusion diagnosis. Because the disease was already advanced at the time of diagnosis and furthermore time was necessary for diagnostic workup, it caused deterioration of the general condition. As a result, adequate chemotherapy could not be performed. There is a need for further case studies to establish diagnosis and treatment of SBA. [ABSTRACT FROM AUTHOR]

Background: To investigate factors that have an impact on the risk of falls and determine whether radiographic knee osteoarthritis (KOA) is a factor involved in falls independent of knee pain, psychological factors, and physical function. Methods: A cross-sectional analysis was conducted on 1083 subjects for the 2009 Locomotive Syndrome and Health Outcomes in the Aizu Cohort Study (LOHAS). A logistic regression analysis was performed to examine the relationship between radiographic KOA and fall history. Results: Fall history was significantly associated with the severity of knee pain. Compared to subjects with no knee pain, the odds ratio (OR) was 1.53 times higher in the subjects with mild knee pain (95% confidence interval [CI]: 1.04–2.25), 1.69 times higher in those with moderate knee pain (95%CI: 1.03–2.79), and 2.98 times higher in those with severe knee pain (95%CI: 1.67–5.30). In subjects with depression, the OR was 1.91 (95%CI: 1.25–2.92), and in those with decreased mobility, the OR was 1.70 (95%CI: 1.08–2.69). Age, gender, knee crepitus, BMI, OLST, and sleeping pill use were not significantly associated with fall risk. In a multivariate analysis, radiographic KOA severity was not significantly associated with fall risk (OR 0.81, 95%CI 0.44–1.50 in mild OA; OR 1.10, 95%CI 0.57–2.14 in severe OA). Conclusion: Knee pain, decreased mobility, and depression, but not the radiographic KOA severity, were significantly associated with a fall risk. Regardless of the individual's radiographic KOA severity, the risk of falls may be reduced by treating his/her knee pain, mobility problems, and/or psychological factors. [ABSTRACT FROM AUTHOR]

Epidermal growth factor receptor (EGFR)‐tyrosine kinase inhibitors are standard therapeutic agents for non‐small cell lung cancer (NSCLC) patients with major EGFR mutations such as exon 19 deletions and a L858R mutation, whereas treatment strategies for cases with uncommon EGFR mutations remain to be fully established. Here, we report a long‐term (≥20 years from initial diagnosis) NSCLC survivor carrying EGFR L858R and L747V mutations. The patient received gefitinib monotherapy, systemic chemotherapy/chemoimmunotherapy, and local consolidative therapies for oligometastatic lesions, and responded to afatinib rechallenge with a progression‐free survival of 12 months. The current case suggests that afatinib is effective in NSCLC patients with EGFR L858R and L747V mutations and that a therapeutic approach combining appropriately timed systemic therapies with local consolidative therapies for oligometastatic lesions improves long‐term survival. [ABSTRACT FROM AUTHOR]

Objective This study aimed to elucidate the actual activities conducted by public health nurses during their dispatch and their health status during and after dispatch to the three prefectures most severely affected by the Great East Japan Earthquake. Sample A survey request was sent to a total of 2,237 facilities. Of these, 778 facilities returned questionnaires from dispatched public health nurses. Results The participants of this study were 1,570 dispatched health nurses who participated in activities mostly at evacuation centers, followed by evacuees' homes. After dispatch, an earlier postdisaster phase at the start of dispatch was independently associated with poor subjective well-being, low mood, worsened sleep state, and intense fatigue. Work hours per day were associated with poor subjective well-being and intense fatigue after dispatch. Conclusion Results suggest that the factor that most strongly affected the postdispatch health of the nurses was the phase that they began their dispatch. [ABSTRACT FROM AUTHOR]

The adhesion of circulating lymphocytes to the surface of vascular endothelial cells is important for their recruitment from blood to secondary lymphoid organs and to inflammatory sites. CD44 is a key adhesion molecule for this interaction and its ligand-binding ability is tightly regulated. Here we show that the hyaluronan-binding ability of CD44 in T cells is upregulated by the depletion of membrane cholesterol with methyl-β-cyclodextrin (MβCD), which disintegrates lipid rafts, i.e. cholesterol- and sphingolipid-enriched membrane microdomains. Increasing concentrations of MβCD led to a dose-dependent decrease in cellular cholesterol content and to upregulation of hyaluronan binding. Additionally, a cholesterol-binding agent filipin also increased hyaluronan binding. Cholesterol depletion caused CD44 to be dispersed from cholesterol-enriched membrane microdomains. Cholesterol depletion also increased the number of cells undergoing rolling adhesion under physiological flow conditions. Our results suggest that the ligand-binding ability of CD44 is governed by its cholesterol-dependent allocation to membrane microdomains at the cell surface. These findings provide novel insight into the regulation of T cell adhesion under blood flow. [ABSTRACT FROM AUTHOR]

BoneXpert, an automated method for analysis of hand radiographs of children, has recently been developed and validated in European children. It determines Tanner-Whitehouse (TW) and Greulich Pyle (GP) bone ages (BA). The purpose of this work is to validate BoneXpert BA in Japanese children and determine the following two properties of the method: (1) The accuracy of the BA, i.e. the standard deviation from an experienced Japanese TW BA rater. (2) The precision of the BA, i.e. BoneXpert's ability to yield the same BA value on a repeated radiograph. The data consist of two studies: 185 radiographs of 22 normal children followed longitudinally from approximately 7 years to full maturity, and 284 radiographs of 22 patients with growth hormone deficiency treated with growth hormone and gonadotropin-releasing hormone analogue followed from an age of 4-11 years to almost full maturity. All radiographs were rated manually according to the TW-Japan system. BoneXpert processed all images, and the accuracy (SD) of TW-Japan BA was 0.72 years (95% CI 0.68-0.76). The precision error (SD) on a single determination of GP BA was 0.17 years (95% CI 0.15-0.19). It is concluded that BoneXpert performs as well in Japanese children as it does in Caucasian children. This study accomplishes a calibration of BoneXpert to the TW-Japan standard, which performs well for the entire BA range from 4 years up to full maturity. Copyright © 2010 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Abstract: On Earth, it is common to employ laboratory animals such as the nematode Caenorhabditis elegans to help understand human health concerns. Similar studies in Earth orbit should help understand and address the concerns associated with spaceflight. The “International Caenorhabditis elegans Experiment FIRST” (ICE FIRST), was carried out onboard the Dutch Taxiflight in April of 2004 by an international collaboration of laboratories in France, Canada, Japan and the United States. With the exception of a slight movement defect upon return to Earth, the result of altered muscle development, no significant abnormalities were detected in spaceflown C. elegans. Work from Japan revealed apoptosis proceeds normally and work from Canada revealed no significant increase in the rate of mutation. These results suggest that C. elegans can be used to study non-lethal responses to spaceflight and can possibly be developed as a biological sensor. To further our understanding of C. elegans response to spaceflight, we examined the gene transcription response to the 10 days in space using a near full genome microarray analysis. The transcriptional response is consistent with the observed normal developmental timing, apoptosis, DNA repair, and altered muscle development. The genes identified as altered in response to spaceflight are enriched for genes known to be regulated, in C. elegans, in response to altered environmental conditions (Insulin and TGF-β regulated). These results demonstrate C. elegans can be used to study the effects of altered gravity and suggest that C. elegans responds to spaceflight by altering the expression of at least some of the same metabolic genes that are altered in response to differing terrestrial environments. [Copyright &y& Elsevier]

Ultrasound stimulation is a type of mechanical stress, and low-intensity pulsed ultrasound (LIPUS) devices have been used clinically to promote fracture healing. However, it remains unclear which skeletal cells, in particular osteocytes or osteoblasts, primarily respond to LIPUS stimulation and how they contribute to fracture healing. To examine this, we utilized medaka, whose bone lacks osteocytes, and zebrafish, whose bone has osteocytes, as in vivo models. Fracture healing was accelerated by ultrasound stimulation in zebrafish, but not in medaka. To examine the molecular events induced by LIPUS stimulation in osteocytes, we performed RNA sequencing of a murine osteocytic cell line exposed to LIPUS. 179 genes reacted to LIPUS stimulation, and functional cluster analysis identified among them several molecular signatures related to immunity, secretion, and transcription. Notably, most of the isolated transcription-related genes were also modulated by LIPUS in vivo in zebrafish. However, expression levels of early growth response protein 1 and 2 (Egr1, 2), JunB, forkhead box Q1 (FoxQ1), and nuclear factor of activated T cells c1 (NFATc1) were not altered by LIPUS in medaka, suggesting that these genes are key transcriptional regulators of LIPUS-dependent fracture healing via osteocytes. We therefore show that bone-embedded osteocytes are necessary for LIPUS-induced promotion of fracture healing via transcriptional control of target genes, which presumably activates neighboring cells involved in fracture healing processes. [ABSTRACT FROM AUTHOR]

Colony spreading of Flavobacterium johnsoniae is shown to include gliding motility using the cell surface adhesin SprB, and is drastically affected by agar and glucose concentrations. Wild-type (WT) and ΔsprB mutant cells formed nonspreading colonies on soft agar, but spreading dendritic colonies on soft agar containing glucose. In the presence of glucose, an initial cell growth-dependent phase was followed by a secondary SprB-independent, gliding motility-dependent phase. The branching pattern of a ΔsprB colony was less complex than the pattern formed by the WT. Mesoscopic and microstructural information was obtained by atmospheric scanning electron microscopy (ASEM) and transmission EM, respectively. In the growth-dependent phase of WT colonies, dendritic tips spread rapidly by the movement of individual cells. In the following SprB-independent phase, leading tips were extended outwards by the movement of dynamic windmill-like rolling centers, and the lipoproteins were expressed more abundantly. Dark spots in WT cells during the growth-dependent spreading phase were not observed in the SprB-independent phase. Various mutations showed that the lipoproteins and the motility machinery were necessary for SprB-independent spreading. Overall, SprB-independent colony spreading is influenced by the lipoproteins, some of which are involved in the gliding machinery, and medium conditions, which together determine the nutrient-seeking behavior. [ABSTRACT FROM AUTHOR]

Objective: To verify the validity of the prediction of oral intake recovery for inpatients with aspiration pneumonia using the Hyodo‐Komagane score. Background: Patients admitted for treatment of aspiration pneumonia sometimes have difficulty in resuming oral intake due to decreased swallowing function. Predicting whether the swallowing function will recover enough to achieve oral ingestion at discharge is an important factor in developing a treatment strategy. No studies have investigated the prediction of oral intake recovery using videoendoscopic examination. Methods: Subjects were 65 patients who were admitted to an acute care hospital for the treatment of aspiration pneumonia. The patients were divided into two groups, the oral feeding group and the tube feeding group, according to their oral intake status at discharge or transfer. Logistic regression analysis was performed using the condition that tube feeding was not required as an objective variable and the items with significant differences between the two groups as explanatory variables. Additionally, the receiver operating characteristic curve was used to identify patients who could take food orally at discharge. Results: The odds ratios for the Hyodo‐Komagane score and the pharyngeal clearance score were 1.485 and 3.379, respectively. When the cut‐off values of the Hyodo‐Komagane score and the pharyngeal clearance score were 6 and 1, the sensitivity was 0.88 and 0.91, and the specificity was 0.64 and 0.70, respectively. Conclusion: The Hyodo‐Komagane score and especially the pharyngeal clearance score are useful indices to predict oral intake recovery for inpatients with aspiration pneumonia. [ABSTRACT FROM AUTHOR]

The malformation and disordered remodeling of bones induce various diseases, including osteoporosis. We have developed atmospheric SEM (ASEM) to directly observe aldehyde-fixed bone tissue immersed in radical scavenger buffer without thin sectioning. The short procedure realized the observation of bone mineralization surrounded by many cells and matrices in natural aqueous buffer, decreasing the risk of changes. In osteoblast primary cultures, mineralization was visible without staining. Correlative energy dispersive X-ray spectrometry indicated the formation of calcium phosphate mineral. Fixed bone was sectioned, and the section surface was inspected by ASEM. Mineralized trabeculae of talus spongy bone were directly visible. Associated large and small cells were revealed by phosphotungstic acid staining, suggesting remodeling by bone-absorbing osteoclasts and bone-rebuilding osteoblasts. In tibia, cortical bone layer including dense grains, was bordered by many cells with protrusions. Tissue immuno-EM performed in solution for the first time and anti-cathepsin-K antibody, successfully identified osteoclasts in femur spongy bone. A microfluidics chamber fabricated on the silicon nitride film window of an ASEM dish allowed mineralization to be monitored in vitro; calcium phosphate crystals as small as 50 nm were imaged. ASEM is expected to be widely applied to study bio-mineralization and bone-remodeling, and to help diagnose bone-related diseases. [ABSTRACT FROM AUTHOR]

Highlights • ASEM of primary culture neurons and their networks in aqueous liquid. • Imaging of fine structures, 50 nm buttons and spines, of synapses in solution. • Line of mitochondria imaged on neurite backbone including microtubule. • ASEM visualized fatty liver tissues of ob/ob type 2 diabetes model mouse. • Osmium tetroxide staining revealed lipid droplets pressing the liver sinusoids. Abstract Neural networking, including axon targeting and synapse formation, is the basis of various brain functions, including memory and learning. Diabetes-mellitus affects peripheral nerves and is known to cause fatty liver disease. Electron microscopy (EM) provides the resolution required to observe changes in fine subcellular structures caused by such physiological and pathological processes, but samples are observed in vacuum. Environmental capsule EM can directly observe cells in a more natural aqueous environment, but the size-limited capsules restrict cell culturability. The recently developed atmospheric scanning electron microscope (ASEM) has an open, 35 mm sample dish, allowing the culture of primary cells, including neurons, on the electron-transparent film window fabricated in its base. The system's inverted scanning electron microscope observes aldehyde-fixed cells or tissues from below through the silicon nitride film; the optical microscope located above allows direct correlation of fluorescence labeling. To observe fixed biological samples, damage due to low dose electron radiation is minimized in three ways. First, knock on damage that pushes out atoms is decreased by the low accelerating voltage of 10–30 kV. Second, increased radical generation due to the decreased acceleration voltage is countered by the addition of a radical scavenger, glucose or ascorbic acid, to the sample solution. Third, the large volume (max. 2 ml) of aqueous buffer surrounding the sample has a high specific heat capacity, minimizing the temperature increase caused by irradiation. Using ASEM, we have developed protocols for heavy metal staining in solution to selectively visualize intracellular structures. Various EM staining methods served as a starting point. Uranyl acetate preferably stains proteins and nucleic acid, and prior tannic acid treatment enhances membranes. Osmium tetroxide is suggested to enhance lipids, especially oil droplets. Imaging primary-culture neurons stained with platinum blue or uranyl acetate revealed growth cones, synapses, and 50–500 nm spines, together with neurite backbones and their associated structures. Correlative microscopy with immuno-fluorescence labeling suggested that these were mainly microtubule associated objects; some showed signs of a fission process and were, thus, possibly mitochondria. Liver tissue excised from the ob/ob type 2 diabetes model mouse, was stained with osmium tetroxide and observed using ASEM. Swollen bright balls occupied a large area of the cytoplasm and could be distinguished from vacuoles, suggesting that they are oil droplets. In some of the images, oil-like droplets were pressing surrounding structures, including sinusoids, significant for blood circulation in the liver. Based on these studies, ASEM combined with metal staining methods promises to allow the study of various mesoscopic-scale phenomena of cells and tissues immersed in natural aqueous environment in the near future. The quick nature of ASEM could facilitate not only the precise imaging for neuroscience but also the diagnosis of fatty liver disease and related diseases. [ABSTRACT FROM AUTHOR]

Purpose: To assess the predictive ability of the maximum chemiluminescence intensity (CImax) for severe neutropenia (SN) during neoadjuvant chemo(radio)therapy [NAC(RT)] in patients with advanced pancreatic or biliary tract cancer.Methods: Clinicopathological variables and blood test data before NAC(RT) were evaluated in 64 patients with advanced pancreatic or biliary tract cancer who received gemcitabine plus tegafur/gimeracil/oteracil as NAC(RT).Results: Thirty-nine patients (60.9%) developed Grade 3-4 SN. The median time between commencing NAC(RT) and the onset of SN was 15 (range 10-36) days. SN occurred during the NAC period, not the RT period. The CImax, neutrophil count, serum interleukin-6 level, C-reactive protein level, complement C3 titer, serum complement titer, and 50.0% hemolytic unit of complement before NAC(RT) were significantly lower in patients with SN than in those without SN (P < 0.05). Multivariate analysis confirmed the CImax to be the sole independent predictor of SN (P < 0.05). The optimal threshold for the CImax was 46,000 RLU/s. The sensitivity and specificity were 46.2% and 80.0%, respectively. Majority of the patients (81.8%) with a low CImax before NAC(RT) experienced SN during NAC(RT).Conclusions: CImax before NAC(RT) predicts SN during NAC(RT) in patients with advanced pancreatic or biliary tract cancer. [ABSTRACT FROM AUTHOR]

This study aimed to identify trends in frequency, serotype, and antimicrobial susceptibility of Streptococcus pneumoniae and Haemophilus influenzae isolated from middle ear fluid specimens of children aged≤15 years (mean, 2 years), before and after the introduction of the 7-valent pneumococcal conjugate vaccine (PCV7) and the H. influenzae type b vaccine, at a pediatric facility in Japan. Sixty-six S. pneumoniae and 88 H. influenzae strains were isolated from 820 middle ear fluid samples. Serotyping and antimicrobial susceptibility testing were performed. The study time-frame was divided into period 1 (2007–2010) and period 2 (2011–2014), according to the availability of vaccine public funding. The S. pneumoniae detection rate decreased from 9.6% in period 1–6.1% in period 2 (p = 0.042). PCV7 serotypes decreased from 56.8% to 9.1% (p = 0.0002). No significant change was observed for the 13-valent pneumococcal conjugate vaccine (PCV13) serotypes: 72.7% in period 1 and 59.1% in period 2. Penicillin-resistant strains (penicillin G-MIC ≥2 μg/mL) decreased from 25% to 4.5% (p = 0.038). Detection rates for H. influenzae did not change significantly: 10.3% in period 1 and 11.3% in period 2. Serotypes were mostly non-typeable: 97.9% in period 1 and 90.2% in period 2, and only one serotype b strain was isolated in each period. The frequency of ampicillin-resistant strains (MIC ≥4 μg/mL) did not change. These results show a preventative effect of PCV7 on otitis media due to S. pneumoniae . PCV7 was replaced with PCV13 in 2013 in Japan; therefore, a further decrease in pneumococcal otitis media is anticipated in the future. [ABSTRACT FROM AUTHOR]

Background: The Great East Japan Earthquake and subsequent tsunami devastated the northeastern part of Japan. Low back pain is thought to increase after a natural disaster and is related to various factors. The aim of this study was to examine the influencing factors of "Living environment" and "Subjective economic hardship" on new-onset of low back pain in the chronic phase for the survivors of the earthquake evaluated by a self-report questionnaire.Methods: A panel study was conducted with the Great East Japan Earthquake survivors at 2 and 3 years after the disaster. New-onset of low back pain was defined as low back pain absent at the 1st period (2 years after the earthquake) and present at the 2nd period (3 years after the earthquake). Living environment was divided into 4 categories (1. Living in the same house as before the earthquake, 2. Living in a prefabricated house, 3. Living in a new house, 4. Others: Living in an apartment, house of relatives or acquaintance). Subjective economic hardship was obtained using the following self-report question: "How do you feel about the current economic situation of your household?" The response alternatives were "Normal", "A little bit hard", "Hard", and "Very hard". A univariate and multivariate logistic regression models were used.Results: 1357 survivors consented to join this study. There was no significant association between new-onset of low back pain and living environment. There was significant association between new-onset of low back pain and "A little hard" (OR = 1.6, 95% CI = 1.07-2.40), "Hard" (OR = 2.2, 95% CI = 1.56-3.74), and "Very hard" (OR = 3.19, 95% CI = 1.84-5.53) in subjective economic hardship.Conclusions: Subjective economic hardship was significantly associated with new-onset of low back pain in the chronic phase for survivors of the Great East Japan Earthquake. [ABSTRACT FROM AUTHOR]

Background: The Great East Japan Earthquake and devastating Tsunami caused irreparable damage on the northeastern coast of Japan. This study aimed to examine the influencing factors of "Living environment" and "Working status" on low back pain for the survivors of the earthquake evaluated by a self-report questionnaire.Methods: Between 2011 and 2013, survivors replied to the self-report questionnaire, and 986 people consented to join this study. The living environment was divided into 3 categories (1. Living in the same house as before the earthquake, 2. Living in a safe shelter or temporary small house, 3. Living in a house of relatives or apartment house) and working status was divided into 5 categories (1. Unemployed before the earthquake, 2. Unemployed after the earthquake, 3. Decrease in income, 4. Different occupation after the earthquake, 5. The same occupation as before the earthquake). Age, gender, living areas, past history of arthritis, arthropathy, osteoporosis, sleep disturbance, psychological distress, and economic status were considered as confounding factors. Generalized estimating regression models with logit link function were used because outcome variables are repeatedly measured and binomial. We evaluated the correlation between the presence/severity of low back pain over time and housing status/working status at 1 year after the earthquake.Results: There were no significant differences between age, gender, living areas, working status, or living environment before or after the earthquake. There was no significant difference in the risk of having low back pain in living environment or gender. There was significant difference in the risk of having low back pain in those with "Decrease in income" (OR = 1.93, 95% CI = 1.23-3.03) and "The same occupation as before the earthquake" (OR = 1.67, 95% CI = 1.1-2.52).Conclusions: Though living environment has little effect, "Decrease in income" and "The same occupation as before the earthquake" have strong influences on low back pain. [ABSTRACT FROM AUTHOR]

Reports on two cases of insulin requiring diabetic patients accompanied by a hallucinatory-delusional state and diagnosed as having schizophrenia. Medical history of the patients; Results of the neurological examination conducted; Improvements observed on the patients after treatment.

Purpose: To evaluate the relationship between skin autofluorescence (SAF), which reflects the accumulation of advanced glycation end products (AGEs), and the severity of diabetic retinopathy (DR) in patients with type 2 diabetes. Methods: Sixty-seven eyes of 67 patients with type 2 diabetes were enrolled. Sixty-seven age-matched non-diabetic subjects served as controls. Diabetic patients were classified by the severity of their DR: no DR (NDR), non-proliferative DR (NPDR), and proliferative DR (PDR). SAF was measured with an autofluorescence reader. Results: SAF in the diabetes patients was significantly higher than in the controls (median 2.5 (interquartile range 2.3-2.7) and 1.8 (1.6-2.3) arbitrary unit (AU), respectively, p < 0.001). There was a statistically significant increase in SAF along with the increasing severity of DR (from NDR to NPDR: p = 0.034; NPDR to PDR: p < 0.01). Logistic regression analysis revealed that SAF (OR, 17.2; p < 0.05) was an independent factor indicating the presence of PDR. Conclusions: SAF has an independent relationship with PDR in patients with type 2 diabetes. SAF measurement with an autofluorescence reader is a non-invasive way to assess the risk of DR. SAF may, therefore, be a surrogate marker candidate for the non-invasive evaluation of DR. [ABSTRACT FROM AUTHOR]

Abstract: Correlative light-electron microscopy of cells in a natural environment of aqueous liquid facilitates high-throughput observation of protein complex formation. ASEM allows the inverted SEM to observe the wet sample from below, while an optical microscope observes it from above quasi-simultaneously. The disposable ASEM dish with a silicon nitride (SiN) film window can be coated variously to realize the primary-culture of substrate-sensitive cells in a few milliliters of culture medium in a stable incubator environment. Neuron differentiation, neural networking, proplatelet-formation and phagocytosis were captured by optical or fluorescence microscopy, and imaged at high resolution by gold-labeled immuno-ASEM with/without metal staining. Fas expression on the cell surface was visualized, correlated to the spatial distribution of F-actin. Axonal partitioning was studied using primary-culture neurons, and presynaptic induction by GluRδ2-N-terminus-linked fluorescent magnetic beads was correlated to the presynaptic-marker Bassoon. Further, megakaryocytes secreting proplatelets were captured, and P-selectins with adherence activity were localized to some of the granules present by immuno-ASEM. The phagocytosis of lactic acid bacteria by dendritic cells was also imaged. Based on these studies, ASEM correlative microscopy promises to allow the study of various mesoscopic-scale dynamics in the near future. [Copyright &y& Elsevier]

tRNase ZL-utilizing efficacious gene silencing (TRUE gene silencing) is a newly developed technology to suppress mammalian gene expression. TRUE gene silencing works on the basis of a unique enzymatic property of mammalian tRNase ZL, which is that it can recognize a pre-tRNA-like or micro-pre-tRNA-like complex formed between target RNA and artificial small guide RNA (sgRNA) and can cleave any target RNA at any desired site. There are four types of sgRNA, 5'-half-tRNA, RNA heptamer, hook RNA, and ∼14-nt linear RNA. Here we show that a 14-nt linear-type sgRNA against human miR-16 can guide tRNase ZL cleavage of miR-16 in vitro and can downregulate the miR-16 level in HEK293 cells. We also demonstrate that the 14-nt sgRNA can be efficiently taken up without any transfection reagents by living cells and can exist stably in there for at least 24 hours. The naked 14-nt sgRNA significantly reduced the miR-16 level in HEK293 and HL60 cells. Three other naked 14-nt sgRNAs against miR-142-3p, miR-206, and miR-19a/b are also shown to downregulate the respective miRNA levels in various mammalian cell lines. Our observations suggest that in general we can eliminate a specific cellular miRNA at least by ∼50% by using a naked 14-nt sgRNA on the basis of TRUE gene silencing. [ABSTRACT FROM AUTHOR]

The incidence rate after a colonoscopic polypectomy includes the true incidence rate of new polyp formation and miss rate of polyps at the initial colonoscopy. It is therefore important to assess accurate incidence rates of polyps as well as those of missing polyps with colocoscopy.Six hundred and eighty-eight patients who underwent total colonoscopy twice within 30 days were investigated to determine the miss rates of neoplastic polyps under colonoscopy. The cumulative incidence rates of neoplasm were evaluated by the Kaplan-Meier method in another series of investigations on 864 patients who underwent surveillance total colonoscopy over 31 days.In 157 (22.8%) patients out of 688, 200 neoplastic lesions (all adenomas) were missed during the first examination. The miss rate was 16% for adenomas≤ 5 mm and 2% for adenomas> 10 mm. The miss rates of patients with 0, 1, 2, or> 2 neoplastic polyps on the first examination was 5.3%, 19.6%, 23.5% and 35.5%, respectively. One year cumulative incidence rates of patients with 1, 2, or> 2 neoplastic polyps on the first examination were 18.4%, 21.1% and 34.2%, respectively.Detection of multiple neoplasms on index colonoscopy predicts a high miss rate and cumulative incidence rate on surveillance colonoscopy. A substantial proportion of neoplasms detected at nearly 1-year postpolypectomy surveillance colonoscopy were missed at the initial total colonoscopy. [ABSTRACT FROM AUTHOR]

Abstract We examined the relations between cognitive function and age and education in the normal elderly population. As per the community-based stroke, dementia, and bed confinement pre-vention in the town of Tajiri, neuropsychological assessments, including the Cognitive Ability Screening Instrument (CASI), were performed for 99 randomly selected normal elderly subjects. We assessed the frontal function (working memory, word fluency, Trail-Making Tests, CASI subitems of list-generating fluency, attention, and concentration/mental manipulation), language function (proverbs, CASI subitem language), non-language function (the digit symbol test of the Wechsler Adult Intelligence Scale-Revised (WAIS-R), CASI subitem visual construction), memory (Alzheimer's Disease Assessment Scale recall/recognition, story recall, CASI subitems short and long-term memory, the Rey-Osterrieth Complex Figure Test), and the global function (CASI subitems orientation and abstraction and judgment). We found that the only test affected by age was the digit symbol test of the WAIS-R. The effects of education were distributed among various tests. There was a significant correlation between age and the frontal lobe atrophy in the lower educated group. The present findings suggest that cognitive function is spared by the aging process itself and dementia should be considered as age-related, not aging-related disorders, and that education might have a protective effect on cognitive change, supporting the reserve hypothesis. [ABSTRACT FROM AUTHOR]

Flavobacterium johnsoniae forms a thin spreading colony on nutrient-poor agar using gliding motility. As reported in the first paper, WT cells in the colony were sparsely embedded in self-produced extracellular polymeric matrix (EPM), while sprB cells were densely packed in immature biofilm with less matrix. The colony surface is critical for antibiotic resistance and cell survival. We have now developed the Grid Stamp-Peel method whereby the colony surface is attached to a TEM grid for negative-staining microscopy. The images showed that the top of the spreading convex WT colonies was covered by EPM with few interspersed cells. Cells exposed near the colony edge made head-to-tail and/or side-to-side contact and sometimes connected via thin filaments. Nonspreading sprB and gldG and gldK colonies had a more uniform upper surface covered by different EPMs including vesicles and filaments. The EPM of sprB, gldG, and WT colonies contained filaments ~2 nm and ~5 nm in diameter; gldK colonies did not include the latter. Every cell near the edge of WT colonies had one or two dark spots, while cells inside WT colonies and cells in SprB-, GldG-, or GldK-deficient colonies did not. Together, our results suggest that the colony surface structure depends on the capability to expand biofilm. [ABSTRACT FROM AUTHOR]

The Gram-negative bacterium Flavobacterium johnsoniae employs gliding motility to move rapidly over solid surfaces. Gliding involves the movement of the adhesin SprB along the cell surface. F. johnsoniae spreads on nutrient-poor 1% agar-PY2, forming a thin film-like colony. We used electron microscopy and time-lapse fluorescence microscopy to investigate the structure of colonies formed by wild-type (WT) F. johnsoniae and by the sprB mutant (ΔsprB). In both cases, the bacteria were buried in the extracellular polymeric matrix (EPM) covering the top of the colony. In the spreading WT colonies, the EPM included a thick fiber framework and vesicles, revealing the formation of a biofilm, which is probably required for the spreading movement. Specific paths that were followed by bacterial clusters were observed at the leading edge of colonies, and abundant vesicle secretion and subsequent matrix formation were suggested. EPM-free channels were formed in upward biofilm protrusions, probably for cell migration. In the nonspreading ΔsprB colonies, cells were tightly packed in layers and the intercellular space was occupied by less matrix, indicating immature biofilm. This result suggests that SprB is not necessary for biofilm formation. We conclude that F. johnsoniae cells use gliding motility to spread and maturate biofilms. [ABSTRACT FROM AUTHOR]

We developed a liquid-phase synthesis method for Pd-based nanostructure, in which Pd dissolved in dimethyl sulfoxide (DMSO) solutions was precipitated using acid aqueous solution. In the development of the method, in situ monitoring using atmospheric scanning electron microscopy (ASEM) revealed that three-dimensional (3D) Pd-based nanonetworks were deformed to micrometer-size particles possibly by the surface tension of the solutions during the drying process. To avoid surface tension, critical point drying was employed to dry the Pd-based precipitates. By combining ASEM monitoring with critical point drying, the synthesis parameters were optimized, resulting in the formation of lacelike delicate nanonetworks using citric acid aqueous solutions. Precipitation using HCl acid aqueous solutions allowed formation of 500-nm diameter nanorings connected by nanowires. The 3D nanostructure formation was controllable and modifiable into various shapes using different concentrations of the Pd and Cl ions as the parameters. [ABSTRACT FROM AUTHOR]

Melanoma is a highly aggressive cancer with a propensity for brain metastases. These can be treated by radiotherapy, but the radiation-resistant nature of melanoma makes the prognosis for melanoma patients with brain metastases poor. Previously, we demonstrated that treatment of mice with subcutaneous melanoma with 5-aminolevurinic acid (5-ALA) and X-rays in combination, ("radiodynamic therapy (RDT)"), instead of with 5-ALA and laser beams ("photodynamic therapy"), improved tumor suppression in vivo. Here, using the B16-Luc melanoma brain metastasis model, we demonstrate that 5-ALA RDT effectively treats brain metastasis. We also studied how 5-ALA RDT damages cells in vitro using a B16 melanoma culture. Cell culture preincubated with 5-ALA alone increased intracellular photosensitizer protoporphyrin IX. On X-ray irradiation, the cells enhanced their ∙OH radical generation, which subsequently induced γH2AX, a marker of DNA double-strand breaks in their nuclei, but decreased mitochondrial membrane potential. After two days, the cell cycle was arrested. When 5-ALA RDT was applied to the brain melanoma metastasis model in vivo, suppression of tumor growth was indicated. Therapeutic efficacy in melanoma treatment has recently been improved by molecular targeted drugs and immune checkpoint inhibitors. Treatment with these drugs is now expected to be combined with 5-ALA RDT to further improve therapeutic efficacy. [ABSTRACT FROM AUTHOR]