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Protein-losing enteropathy (PLE) is a rare syndrome characterized by hypoproteinemia due to gastrointestinal (GI) protein loss. Primary intestinal follicular lymphoma (PIFL), a specific variant of follicular lymphoma with essential only GI involvement, has not been reported as an etiology of PLE. We herein report a case of PLE complicated with PIFL that was successfully treated with rituximab, resulting in rapid improvement of PLE and a complete response of PIFL. Macroscopic findings of ulcerative lesions with diffuse involvement, which were precisely described by capsule and double-balloon enteroscopy at the diagnosis, also improved following the treatment. This case provides a clue suggesting factors that promote PLE in PIFL.

DNA immunization has been used to study vaccination methods and for production of specific antibodies. The present study aimed to apply DNA immunization to prepare specific IgYs, which react against rabies virus N protein (RV-N) and can be used to research and diagnose rabies virus. The DNA sequence of RV-N was ligated into a pcDNA 3.1 plasmid for constructing pcDNA-N. Eight hens were divided into four groups. Group 1 comprised the control group (non-immunized). In Groups 2, 3, and 4, hens were injected intramuscularly with pcDNA-N (400 µg/hen). Eight injections were administered every other week. From the 4th week, an adjuvant was injected in addition to pcDNA-N. Freund's complete adjuvant (FCA) and

Several region-specific vegetable varieties in Japan have an extensive cultivation history. Distinct tastes and aromas make them suitable for use in spices and pickles, and varieties with a high co...

We previously reported a novel diagnostic method using follicle-sinus complexes (FSCs) in the muzzle skin for postmortem diagnosis of rabies in dogs. However, whether this method works in other animal species remains unclear. Here, FSCs were collected from a wolf, a red fox, 2 bats, and a cat, and examined for the presence of viral antigen, viral mRNA, and viral particles. Viral antigen and viral mRNA were confirmed in Merkel cells (MCs) in FSCs of all species. Electron microscopy performed using only samples from wolf and cat confirmed viral particles in MCs of FSCs. These results suggested that this novel diagnostic method using FSCs might be useful for detection of rabies not only in domestic but also wild animals.

Rabies is a highly neurotropic disease caused by rabies lyssavirus (RABV). Human rabies vaccines exist for pre- and postexposure prophylaxis; however, after clinical symptoms appear, the disease has an ∼100% mortality rate with no effective treatments available. In our previous study, mouse neuroblastoma cells transfected with a plasmid coding one clone of a single-chain variable fragment (scFv), scFv-P19, against RABV phosphoprotein (RABV-P) derived from an scFv phage-display library, before infection, exhibited reduced viral propagation after infection with the RABV-fixed strain, CVS11. In this study, we conducted epitope mapping of scFv-P19 through indirect fluorescent assay and Western blotting analysis against full-length and N- or C-terminal truncated RABV-P. Our results suggest that scFv-P19 targets a portion containing amino acids 47-52 at the N-terminus, which partially overlaps with the N-terminal nuclear export sequences. This provides insights into the underlying mechanism associated with inhibition of RABV by scFv-P19, while allowing for the design of additional scFv-based therapeutic studies for RABV by integrating appropriate delivery and application systems. Furthermore, the results of this study suggest that scFv-P19 may serve as an effective tool for investigating nuclear trafficking of RABV-P to explore the roles of RABV-P isoforms in rabies pathogenesis.

Exercise is a key intervention for improving older adults’ physical function and life expectancy. Here, we investigated a short-term intervention program designed to improve the physical functioning of elderly adults in a community-dwelling setting. We examined the effect of a 5-week combined exercise and education program on the physical function, social engagement, mobility performance, and fear of falling in 42 subjects older than 65 years. Eleven subjects dropped out. There was significant improvement in the 30-second chair stand test ( p

Pancreatic cancer is a malignant neoplasm with high invasiveness and poor prognosis. In a previous study, a highly invasive pancreatic cancer cell line was established and found to feature enhanced interleukin-32 (IL-32) expression. However, whether IL-32 promotes the invasiveness by enhancing or suppressing the expression of IL-32 through regulating downstream molecules was unclear. To investigate the effect of IL-32, cells were established with high levels of expression or downregulated IL-32; their invasive ability was measured using a real-time measurement system and the expression of some candidate downstream molecules involved in invasion was evaluated in the two cell types. The morphological changes in both cell types and the localization of IL-32 expression in pancreatic cancer tissues were studied using immunohistochemistry. Among the several splice variants of IL-32, cells transfected with the ε isoform had increased invasiveness, whereas the IL-32-suppressed cells had reduced invasiveness. Several downstream molecules, whose expression was changed in the two cell types, were monitored. Notably, changes of E-cadherin, CLDN1, CD44, CTGF and Wnt were documented. The morphologies of the two cell types differed from the original cell line. Immunohistochemically, the expression of IL-32 was observed only in tumor cells and not in normal pancreatic cells. In conclusion, IL-32 was found to promote the invasiveness of pancreatic cancer cells by regulating downstream molecules.

A 32-year-old man, who visited Japan from the Philippines in 2020, was diagnosed with rabies, the first reported case in Japan since 2006. This is the fourth imported case of rabies since 1957; one case in 1970 was imported from Nepal and two in 2006 were imported from the Philippines.

BackgroundOvarian tumors are predominantly of epithelial origin and are currently divided into three groups: Adenoma, borderline tumor, and adenocarcinoma by the presence or absence of invasion and cellular atypia. However, it is difficult to assess invasion, so a more objective biomarker would be desirable. Laminin-5 (Lam5) is a protein that constitutes the extracellular matrix of the basement membrane, and it is composed of three short-chain subunits. One of them, the Lam5γ2 chain (Lam5γ2), has been reported to be expressed in some malignant tumors and is suggested to be related to tumor cell invasion. Against this background, we investigated immunohistologically whether the Lam5γ2 would be useful as a marker to predict invasion in ovarian serous and mucinous tumors.MethodsImmunohistochemistry for Lam5γ2 was performed on a total of 80 cases of serous and mucinous tumor adenomas, borderline tumors, and adenocarcinomas, and the differences in the localization of Lam5γ2 expression were observed.ResultsThe basement membrane expression of Lam5γ2 tended to be preserved or had disappeared in half of the adenomas and borderline serous tumors. In all cases of adenocarcinoma, the expression on the basement membrane had disappeared. The frequency of expression in tumor cells increased in the order of adenoma, borderline tumor, adenocarcinoma. In particular, in most adenocarcinoma cases a cytoplasmic expression was observed. The same tendency was noted in mucinous tumors, and the basement membrane expression tended to disappear in the order of adenoma, borderline tumor and adenocarcinoma. Many cases of adenocarcinoma were observed among the tumor cells. In adenocarcinoma cases of both types of tumor, many tumor cells showed prominent cytoplasmic expression particularly in the microinvasive foci and in the invasive front.ConclusionsThe disappearance of Lam5γ2 from the basement membrane and its aberrant expression in serous and mucinous tumor cells may serve as a phenotype for invasiveness.

AIMS This study investigated the relationship between the differentiation of tumour cells into crypts, which is determined by cell differentiation into Paneth and neuroendocrine cells, and tumour infiltration in gastric dysplasia. METHODS AND RESULTS The lesions were endoscopically biopsied low-grade dysplasia (LGD), endoscopically resected high-grade dysplasia (HGD), or cancer with submucosal invasion. LGD (n = 32) displayed crypt differentiation across the entire width of the tumour in all cases. Crypt differentiation was identified as a characteristic of tumours with low biological malignancy. HGD (n = 40) included tumours with a mixture of areas with and without crypt differentiation (n = 25) and tumours with crypt differentiation throughout the entire width (n = 15). Of the cancers with submucosal invasion (n = 30), the morphological progression of HGD area with crypt differentiation, HGD area without crypt differentiation and invasive cancer without crypt differentiation was confirmed for 23 samples. In two lesions, invasive cancer without crypt differentiation developed from HGD without crypt differentiation throughout the tumour width. In five samples, well-differentiated tubular adenocarcinoma with crypt differentiation developed from HGD with crypt differentiation and invaded with lamina propria-like stroma. CONCLUSIONS Loss of crypt differentiation could be an objective indicator of infiltration in the progression of HGD to invasive cancer. The invasive potential of dysplasia depends on the presence or absence of crypt differentiation.

A novel indirect fluorescent antibody test (IFAT) for detection of IgM against Nipah virus (NiV) was developed using HeLa 229 cells expressing recombinant NiV nucleocapsid protein (NiV-N). The NiV IFAT was evaluated using three panels of sera: a) experimentally produced sera from NiV-N-immunized/pre-immunized macaques, b) post-infection human sera associated with a Nipah disease outbreak in the Philippines in 2014, and c) human sera from a non-exposed Malaysian population. Immunized macaque sera showed a characteristic granular staining pattern of the NiV-N expressed antigen in HeLa 229 cells, which was readily distinguished from negative-binding results of the pre-immunized macaque sera. The IgM antibody titers in sequential serum samples (n = 7) obtained from three Nipah patients correlated well with previously published results using conventional IgM capture ELISA and SNT serology. The 90 human serum samples from unexposed persons were unreactive by IFAT. The IFAT utilizing NiV-N-expressing HeLa 229 cells to detect IgM antibody in an early stage of NiV infection is an effective approach, which could be utilized readily in local laboratories to complement other capabilities in NiV-affected countries.

Background: Cetuximab is a powerful anti-neoplastic agent that can inhibit cell growth in oral squamous cell carcinomas (OSCCs). Unfortunately, there are cases with unfavorable outcomes. Because few studies have focused on the combined effects of cetuximab and histone deacetylase (HDAC) inhibitors, we aimed to evaluate the antitumor effect of cetuximab in combination with HDAC inhibitors in human OSCC cell lines, and investigate the mechanism of apoptosis enhancing activity thereof.Methods: We used human OSCC cell lines treated with cetuximab (500 mg/ml) and several HDAC inhibitors. The WST assay and ApoToxGlo™ Triplex Assay determined cell survival. We employed the TdT-mediated dUTP-biotin nick end labeling method to detect apoptosis. We used western blotting to examine the histone acetylation status, ER stress markers, and epidermal growth factor receptor (EGFR) signaling pathways.Results: Cetuximab in combination with 4-phenyl butyric acid (4-PBA) remarkably decreases cell growth in vitro. In addition, the combined treatment resulted in increased EGFR mRNA expression, and it promoted the activation of ERK. The combination treatment induced apoptosis at a significantly higher frequency than did either agent alone.Conclusions: The combination of cetuximab and 4-PBA is more effective against human OSCC cells than either agent alone, suggesting a potential clinical applicability of combination treatment in OSCC therapies.

The area of Kanto-Koushinetsu occupies 16% (63,041 km2) of the land area of Japan. This region includes the Tokyo metropolis and the prefectures of Ibaraki, Tochigi, Gunma, Saitama, Chiba, Kanagawa, Yamanashi, Nagano, Niigata. Significant depletion in agricultural land has been progressed from 1972 to 2011 due to the population concentration in the urban area of Tokyo metropolis and the prefectures of Saitama, Chiba, Kanagawa. The area around the population concentration zone plays an important role in supplying vegetables, and the national share of items is 33.8% for vegetables and 25.6% for a flowering plant, and these values are the top of the Japan. In the Kanto region, the plain area occupies a large area, and agricultural activities are being developed under good water use and the existence of a populated area. The characteristics of geology, topography, climatic conditions and soil characteristics and the effects of volcanic ash soil in this area are described. The relationship between the recent agricultural production of Kanto-Koshinetsu area and the soil characteristics of each area was clarified. On the other hand, anthropogenic soils are increasing in Tokyo metropolis and problems in current soil classification system emerged from these soils has been pointed out. As a topic, an action on restoration of habitat for the reintroduction of the crested ibis in Sado Island (Sado City, Niigata Prefecture) for selected as Globally Important Agricultural Heritage System (GIAHS) was introduced. Furthermore, examples of agricultural production and soil environment conservation were introduced.

Extramammary Paget's disease (EMPD) is a rare malignant skin neoplasm characterized by intraepidermal proliferation of tumor cells. The tumor cells of EMPD may sometimes invade into the dermis or metastasize into the regional lymph nodes. Several studies have proposed mechanisms underlying the increased invasiveness of EMPD; however, molecular markers indicating invasiveness have yet to be well characterized. Laminin-5 (Lam-5), a heterotrimer composed of three chains (α3, β3, and γ2), is a major component of the basement membrane in many tissues. One of the chains, Lam-5 γ2, is a marker of invasion, because it often develops as a monomer in malignant neoplasms. We investigated the expression of Lam-5 γ2 and its role for the invasiveness in EMPD. Paraffin-embedded specimens of EMPD obtained from 36 patients were examined immunohistochemically for Lam-5 γ2. The cases adopted into this study comprised 16 cases of intraepidermal lesions and 20 cases with dermal invasion. The basement membrane seen in normal skin disappeared in one-third of non-invasive cases and in most invasive cases. The disappearance of Lam-5 γ2 in the basement membrane and its cytoplasmic expression was more observed in the invasive cases than non-invasive cases. Expression of Lam-5 γ2 may be a biological marker to predict invasiveness of EMPD.

Compared to tumors of other organs, pancreatic cancer is highly aggressive; with one of its biological features being that, despite a prominent fibrotic stroma, there is remarkable infiltration of tumor cells. This characteristic is considered to be the main reason for the poor prognosis of patients with pancreatic cancer. Therefore, in order to elucidate the factors that contribute to this high invasiveness, a selective invasion method was used to establish four highly invasive subclones from six human pancreatic cancer cell lines. The results demonstrated that two cell lines did not exhibit enhanced invasiveness. Microarray analysis revealed that, in the highly invasive cell lines, several genes were expressed at high levels, compared with the original cell lines. These highly expressed genes were recognized only in highly invasive cells. Among them, IL-32 was most strongly upregulated in the highly invasive cells, compared with cells with a low invasive potential, as well as the original cells. RT-qPCR and western blot analysis confirmed the high levels of expression of IL-32 in highly invasive cells at the RNA and protein levels. In addition, immunohistochemical analysis of resected surgical materials revealed that the tumor cells expressed IL-32 and, in particular, many IL-32 positive cells were seen at the invasive front of the tumor tissue. IL-32 is a cytokine that is widely involved in the development of cancer and has recently received considerable attention. This cytokine has multiple splice variants and shows a wide variety of behaviors, depending on the tumor type and primary organ. Although some hypotheses have been proposed to explain the activity of IL-32, a unified view has not been agreed. In the present study, through the establishment of highly invasive cells from pancreatic cancer and a comprehensive gene analysis, it is suggested that IL-32 may serve an important role as a molecule involved in the invasiveness of this neoplasm.

Recently, we reported that follicle-sinus complexes (FSCs) in the muzzle skin are useful for postmortem diagnosis of rabid dogs. Here, we compared the sensitivity and specificity of detecting the viral antigen in the brain and FSCs of 226 suspected rabid dogs, and assessed whether the FSC harbored the virus genome and particles. The viral antigen was detected in 211 of 226 samples with 100% sensitivity and specificity. Viral RNA and particles were observed in the cytoplasm of Merkel cells (MCs). These results suggest that MCs are targets of virus infection and FSCs are useful material for diagnosing rabies.

Background The complete genome sequences of 44 Bacillus cereus group isolates collected from diverse sources in Japan were analyzed to determine their genetic backgrounds and diversity levels in Japan. Multilocus sequence typing (MLST) and core-genome single-nucleotide polymorphism (SNP) typing data from whole-genome sequences were analyzed to determine genetic diversity levels. Virulence-associated gene profiles were also used to evaluate the genetic backgrounds and relationships among the isolates. Results The 44 B. cereus group isolates, including soil- and animal-derived isolates and isolates recovered from hospitalized patients and food poisoning cases, were genotyped by MLST and core-genome SNP typing. Genetic variation among the isolates was identified by the MLST and core-genome SNP phylogeny comparison against reference strains from countries outside of Japan. Exploratory principal component analysis and nonmetric multidimensional scaling (NMDS) analyses were used to assess the genetic similarities among the isolates using gene presence and absence information and isolate origins as the metadata. A significant correlation was seen between the principal components and the presence of genes encoding hemolysin BL and emetic genetic determinants in B. cereus, and the capsule proteins in B. anthracis. NMDS showed that the cluster of soil isolates overlapped with the cluster comprising animal-derived and clinical isolates. Conclusions Molecular and epidemiological analyses of B. cereus group isolates in Japan suggest that the soil- and animal-derived bacteria from our study are not a significant risk to human health. However, because several of the clinical isolates share close genetic relationships with the environmental isolates, both molecular and epidemiological surveillance studies could be used effectively to estimate virulence in these important pathogens.

In this paper, we propose a trajectory estimation method using sparsely deployed anchor nodes. Whenever mobile nodes encounter each other, they record the encounter time and ID of the encounter node. Moreover, when they encounter anchor nodes, they forward the encounter records to the anchor nodes. The anchor nodes then send the encounter records to a cloud server. In our proposed method, the cloud server estimates the trajectory of the mobile nodes based on the encounter records, the positions of the anchor nodes, and the average moving speed of the mobile nodes. Through simulation experiments, we show the effectiveness of our proposed method.

Background/aim Angiosarcoma is a rare and aggressive soft tissue sarcoma with poor prognosis. Chemotherapy and radiotherapy do not improve the prognosis. SIRT1, a class III histone deacetylase, is up-regulated in many malignant tumours. This study aimed at exploring the role of SIRTl in angiosarcoma. Materials and methods The effect of suppressing SIRT1 expression with siRNA on the proliferation and invasion ability of ISO-HAS-B angiosarcoma cells was investigated. Additionally, SIRT1 expression in tissues from surgical specimens was immunohistochemically evaluated and compared to that from benign tumours. Results Suppression of SIRT1 expression by siRNA resulted in the down-regulation of cell growth, proliferation, migration, and invasion. An immunohistochemical analysis disclosed that SIRT1 expression in angiosarcoma was stronger than that in haemangioma. Conclusion SIRT1 may be involved in the invasive proliferation and malignant transformation of angiosarcoma, and may be considered a future target for angiosarcoma therapy.

Peripheral electrical stimulation (PES) modulates the excitability of the corticospinal tract (CST). This modulation of CST excitability depends on the PES intensity, defined by the amplitude and the width of each pulse, the total pulse number, the stimulation frequency, and the intervention duration. Another key PES parameter is the stimulation pattern; little is known about how PES pattern affects CST excitability, as previous studies did not control other PES parameters. Here, we investigated the effect of the net difference in PES pattern on CST excitability. We use three controlled PESs, intermittent PES (30 Hz) (stimulation trains at 30 Hz with pauses), continuous PES (12 Hz) (constant stimulation at 12 Hz without pauses), and continuous PES (30 Hz) with the same stimulation frequency as the intermittent PES (30 Hz), to compare the effect of the stimulation frequency. The motor evoked potentials (MEPs) and somatosensory evoked potentials (SEPs) of healthy subjects were recorded before and after these three types of PESs in separate sessions. We found that intermittent PES (30 Hz) increased MEP amplitudes, whereas continuous PES (12 and 30 Hz) decreased amplitudes. A significant change in subcortical SEP component occurred during continuous PES (12 and 30 Hz), but not intermittent PES (30 Hz), whereas cortical SEP components showed similar behavior in three types of PESs. We conclude that (1) opposing modulations of CST excitability were induced by the differences in the PES pattern, and (2) these modulations appear to be mediated through different processes in the sensorimotor system. Our findings suggest the possibility that it may be preferable to select the PES pattern in therapeutic interventions based on the putative desired effect and the neural structure being targeted.

A 41-year-old female smoker presented with a vaginal mass. Gynecological examination showed a mass filling the uterine corpus, cervix, and vagina. A total abdominal hysterectomy was performed. Macroscopic findings included a large fragile mass involving the uterine cavity, cervix, and vagina. Histology revealed atypical ducts admixed with solid components consisting of large atypical cells. The initial pathological diagnosis was grade 3 endometrioid adenocarcinoma. The patient was designated as stage II according to the 2008 International Federation of Gynecology and Obstetrics (FIGO) staging. Two years later, two nodules were found in the upper lobe of the left lung, and the patient underwent an upper lobectomy. The masses, which exhibited solid and organoid growth patterns of large atypical cells, had histological characteristics of large cell neuroendocrine carcinoma (LCNEC) of the lung. However, the tumor was immunohistochemically positive for neuroendocrine markers, such as synaptophysin in addition to estrogen receptor and progesterone receptor, and the tumor was negative for thyroid transcription factor-1. These immunohistochemical results were almost identical to those of the solid portions of the uterine carcinoma. The final diagnosis was LCNEC combined with endometrioid adenocarcinoma of the uterine corpus and lung metastasis of the LCNEC component of the endometrial carcinoma. LCNEC often arises in the lung, but it rarely arises in other organs. Some patients with metastatic components exhibited only a LCNEC pattern although the primary tumor was a mixed carcinoma consisting of LCNEC and other histology, like the present case. LCNEC is often poorly differentiated, especially in extrapulmonary primary organ LCNEC. Therefore, pathologists should consider metastatic carcinoma when they encounter lung LCNEC in a patient with a preceding extrapulmonary carcinoma composed of a poorly differentiated component or LCNEC component, and they should clarify tumor immunohistochemical characteristics to confirm the diagnosis.

The coverage of rabies vaccinations has been reported at 70-80% of dogs in annual reports. However, there are still outbreaks of rabies among humans and dogs in Thailand, thus indicating the necessity of ensuring seroprevalence in vaccinated dogs and efficacy of human immunization. A cost effective easy competitive enzyme-linked immunosorbent assay (CEE-cELISA) was developed here for monitoring protective immunity against the rabies virus in human and dog serum samples using monoclonal antibody clone 1-46-12, which recognizes a conformational epitope of the rabies G protein. The ELISA plate is coated with the whole viral antigen from a commercial vaccine. The serotiter measured by the CEE-cELISA and by the gold standard assay (rapid fluorescent focus inhibition test), detecting the neutralizing antibody, showed a strong correlation, with an R value of 0.958 and 0.931 in humans and dogs, respectively. These correlations were detected in the serum samples from humans and dogs at antibody concentrations up to 100 and 10 IU/ml, respectively. This CEE-cELISA could be an alternative assay for evaluating mass rabies vaccination rapidly at a low cost as well as for detecting antirabies antibodies in the serum of not only humans but also other animal species.

Prenylated indole alkaloid okaramines selectively target insect glutamate-gated chloride channels (GluCls). Because of their highly complex structures, including azocine and azetidine rings, total synthesis of okaramine A or B has not been achieved, preventing evaluation of the biological activities of okaramines. Biosynthetic approaches provide alternatives to accessing structurally diverse derivatives and enabling the elucidation of structure-activity relationships. To explore the biosynthetic potential of okaramines, gene knockout experiments of an okaramine-producer fungus were performed. The deletion mutants of the oxygenase genes okaB, okaD, okaE, and okaG provided analogues that were unlikely to be accumulated in the normal biosynthetic process of the wild-type strain. Analysis of the structure-activity relationships of okaramines collected from the fungal cultures revealed that 1,4-dihydroazocine and N-aliphatic group attached to the indole were crucial for GluCl-activating activity. This provided insights into further derivatization of the complex structure of okaramines in order to facilitate the development of new insecticides.

Expression of the protein deacetylase SIRT1 is associated with either poor or favorable prognosis in cancer patients, depending on the cancer type. In head and neck squamous cell carcinoma (HNSCC), SIRT1 expression is associated with favorable prognosis. However, the molecular mechanism underlying the tumor-suppressive function of SIRT1 in HNSCC is unknown. SIRT1 promotes differentiation in epithelial cells; therefore, we investigated whether SIRT1 promotes differentiation in HNSCC cells by studying the correlations between the expression of SIRT1 and several genes implicated in stemness or differentiation in HNSCC-derived cell lines. Our results suggest that SIRT1 does not contribute to differentiation in HNSCC cells. RNA interference-mediated reduction of SIRT1 revealed that SIRT1 supports the expression of TAp63, which has been implicated in tumor suppression, in addition to epithelial differentiation. A positive correlation was observed between SIRT1 and TAp63 expression in HNSCC tissues, as determined by quantitative reverse transcription-polymerase chain reaction analysis of RNA extracted from formalin-fixed paraffin-embedded biopsy samples. Together, these results suggest that although SIRT1 does not regulate differentiation of HNSCC cells, it functions as a tumor suppressor in HNSCC by supporting the transcription of tumor-suppressive TAp63. This finding supports the notion that SIRT1-activating drugs could be useful for the treatment of HNSCC.

In this study, G proteins of the rabies virus (RABV) Kyoto strain were detected in the cytoplasm but not distributed at the cell membrane of mouse neuroblastoma (MNA) cells. G proteins of CVS-26 were detected in both the cell membrane and perinuclear space of MNA cells. We found that N-glycosylation of street RABV G protein by the insertion of the sequon Asn(204) induced the transfer of RABV G proteins to the cell surface membrane. Fixed RABV budding from the plasma membrane has been found to depend not only on G protein but also on other structural proteins such as M protein. However, the differing N-glycosylation of G protein could be associated with the distinct budding and antigenic features of RABV in street and fixed viruses. Our study of the association of N-glycan of G protein at Asn(204) with the transport of RABV G protein to the cell surface membrane contributes to the understanding of the evolution of fixed virus from street virus, which in turn would help for determine the mechanism underlying RABV budding and enhanced host immune responses.

Two Melaleuca species, M. cajuputi and M. bracteata, were compared to identify the factors determining their distinct aluminum (Al) resistance levels. The presence of Al in a liquid culture medium (maximum tested concentration, 2 mM) did not affect the growth of M. cajuputi, but severely inhibited the growth of M. bracteata. The Al content in the roots was 50% higher in Al-sensitive M. bracteata than in Al-resistant M. cajuputi. Al penetration and tissue damage were obvious in the roots of M. bracteata, but only mild in the roots of M. cajuputi. Relatively high levels of fumarate were released by the roots of M. cajuputi, but not by those of M. bracteata. Supplementation of Al-containing liquid media with fumarate resulted in a reduction of Al toxicity on M. bracteata. These results suggest that Al-resistant M. cajuputi releases fumarate from its roots, thereby detoxifying Al.

Clinical trials of histone deacetylase (HDAC) inhibitors as antitumor therapy have been conducted for gastric cancer. Expression of SIRT1, a class III HDAC, is related to poor prognosis in some malignancies. We investigated the correlation between SIRT1 expression and progression and prognosis of gastric cancers comparing with molecules linked to SIRT1 in order to better predict the efficacy of HDAC inhibitors in treating this disease. We evaluated SIRT1 expression by western blot in 51 cases and SIRT1, DBC1, acetylated H4K16 (H4K16Ac), acetylated H3K9 (H3K9Ac), and p53 by immunohistochemistry (IHC) in 557 cases of gastric cancer. Western blotting showed that SIRT1 high expression related with statistics to advanced tumor progression, positive lymphatic invasion, positive venous invasion, and advanced stage but not to poor prognosis. IHC revealed that SIRT1 high expression correlated with worse clinico-pathological prognostic factors as same as in western blotting and related poor prognosis both by univariate and multivariate analyses. By the contrast, DBC1 and H4K16Ac were related to favorable prognostic factors and linked to favorable prognosis by univariate analysis but not by multivariate analysis. H3K16Ac correlated only favorable prognostic factors. Results of p53 were very similar to those of SIRT1. We found that SIRT1 high expression closely correlates with progression and prognosis in gastric cancer patients. And it was also indicated that SIRT1 acts as an oncogene by the results of DBC1, H4K16Ac, and H3K9Ac and might be a target molecule of HDAC inhibitor treatment for gastric cancer patients.

Many attempts to demonstrate the oncogenic role of the JC virus (JCV) have been partially successful in producing brain tumors, either by direct inoculation of JCV into the brain or in transgenic models in rodents. We previously reported the presence of JCV DNA with a relatively high incidence in pulmonary and digestive organs. However, we could not prove the oncogenic role of JCV. We prepared a transgene composed of the K19 promoter, specific to bronchial epithelium with the JCV T-antigen and established transgenic (TG) mice. Pulmonary tumors were detected without any metastasis in 2 out of 15 (13.3%) 16-month-old K19/JCV T-antigen TG mice. Using immunohistochemistry (IHC), these tumors showed JCV T-antigen, p53 and CK 19 expression, but not expression of nuclear and cytoplasmic β-catenin and insulin receptor substrate 1 (IRS1). IHC revealed the same expression pattern as in the bronchial epithelium of the TG mice. One tumor, which was examined with laser capture microdissection and molecular biological tools, demonstrated an EGFR mutation but not a K-ras mutation. We propose that the pulmonary tumors were derived from the JCV T-antigen in a TG mouse model. These findings shed light on pulmonary carcinogenesis.

Aluminum (Al) accumulation and long-distance transport in oil tea (Camellia oleifera Abel.), known to be an Al accumulator, was investigated. The average Al concentration in the embryo of oil tea seeds was 389 mg Al kg(-1) dry weight, which was higher than seeds of other Al accumulators. By partially suppressing leaf transpiration in the field, Al accumulation in leaves was depressed, which clarified the importance of xylem transport to Al accumulation in leaves. However, the effects of xylem transport alone could not sufficiently explain the high Al accumulation in the seasons when the leaf transpiration is weak, which hints the necessity of phloem transport working. Aluminum content in phloem exudates of barks provides another evidence of phloem transport. Images from scanning electron microscopy and energy-dispersive analysis also showed that Al was present in the phloem of oil tea petioles. Aluminum in oil tea could also be redistributed: higher concentrations of Al were found in leaves when Al was supplied to a different leaf of the same plant. In addition, Al was present in newly emerging roots of oil tea seedlings in which all original roots were excised prior to treatment, and a positive correlation existed between Al content in the newly formed roots and that in the leaves. The results using the empty seed coat technique showed that Al unloading via the phloem occurred during seed development. In conclusion, the results demonstrated that Al could be redistributed between leaves, from seeds to leaves, leaves to roots and leaves to seeds, which indicates that Al can be transported via the phloem in oil tea.

† Background and Aims Acidic soils are dominated chemically by more ammonium and more available, so more potentially toxic, aluminium compared with neutral to calcareous soils, which are characterized by more nitrate and less available, so less toxic, aluminium. However, it is not known whether aluminium tolerance and nitrogen source preference are linked in plants. † Methods This question was investigated by comparing the responses of 30 rice (Oryza sativa) varieties (15 subsp. japonica cultivars and 15 subsp. indica cultivars) to aluminium, various ammonium/nitrate ratios and their combinations under acidic solution conditions. † Key Results indica rice plants were generally found to be aluminium-sensitive and nitrate-preferring, while japonica cultivars were aluminium-tolerant and relatively ammonium-preferring. Aluminium tolerance of different rice varieties was significantly negatively correlated with their nitrate preference. Furthermore, aluminium enhanced ammonium-fed rice growth but inhibited nitrate-fed rice growth. † Conclusions The results suggest that aluminium tolerance in rice is antagonistic with nitrate preference and synergistic with ammonium preference under acidic solution conditions. A schematic diagram summarizing the interactions of aluminium and nitrogen in soil‐plant ecosystems is presented and provides a new basis for the integrated management of acidic soils.

A novel antigen-capture sandwich ELISA system targeting the glycoproteins of the henipaviruses Nipah virus (NiV) and Hendra virus (HeV) was developed. Utilizing purified polyclonal antibodies derived from NiV glycoprotein-encoding DNA-immunized rabbits, we established a system that can detect the native antigenic structures of the henipavirus surface glycoproteins using simplified and inexpensive methods. The lowest detection limit against live viruses was achieved for NiV Bangladesh strain, 2.5 × 10(4) TCID(50). Considering the recent emergence of genetic variants of henipaviruses and the resultant problems that arise for PCR-based detection, this system could serve as an alternative rapid diagnostic and detection assay.

Nipah virus (NiV), Paramyxoviridae, Henipavirus, is classified as a biosafety level (BSL) 4 pathogen, along with the closely related Hendra virus (HeV). A novel serum neutralization test was developed for measuring NiV neutralizing antibodies under BSL2 conditions using a recombinant vesicular stomatitis virus (VSV) expressing secreted alkaline phosphatase (SEAP) and pseudotyped with NiV F/G proteins (VSV-NiV-SEAP). A unique characteristic of this novel assay is the ability to obtain neutralization titers by measuring SEAP activity in supernatant using a common ELISA plate reader. This confers a remarkable advantage over the first generation of NiV-pseudotypes expressing green fluorescent protein or luciferase, which require expensive and specific measuring equipment. Using panels of NiV- and HeV-specific sera from various species, the VSV-NiV-SEAP assay demonstrated neutralizing antibody status (positive/negative) consistent with that obtained by conventional live NiV test, and gave higher antibody titers than the latter. Additionally, when screening sixty-six fruit bat sera at one dilution, the VSV-NiV-SEAP assay produced identical results to the live NiV test and only required a very small amount (2μl) of sera. The results suggest that this novel VSV-NiV-SEAP assay is safe, useful for high-throughput screening of sera using an ELISA plate reader, and has high sensitivity and specificity.

The central nervous system (CNS) tissue of mice infected with the CVS-11 strain of rabies virus (RABV) was subjected to gene expression analysis using microarray and canonical pathway analyses. Genes associated with innate immunity as well as inflammatory responses were significantly up-regulated, corroborating with the previous findings obtained using attenuated viruses that did not induce a fatal outcome in infected mice. Histopathological examination showed that neurons in the cerebellum had undergone apoptosis. Although the extent of Fas ligand up-regulation was not so prominent, perforin and granzyme genes were highly expressed in the CNS of mice infected with CVS-11. The presence of perforin and granzymes both in the Purkinje cells and CD3 T lymphocytes strongly suggested that apoptosis of the former cells was induced by the latter cells.

Rabies virus (RABV) is highly neurotropic and causes acute infection of the central nervous system. Death can be averted by prompt post-exposure prophylaxis; however, after clinical symptoms appear, the mortality rate is almost 100% and no reliable treatment is available. In this study, we investigated whether intracellular immunization using single-chain variable fragments (scFvs) against RABV phosphoprotein (RABV-P) could inhibit RABV propagation in neuronal cells. Of four scFv clones derived from an scFv phage-displayed library, scFv-P19 showed extremely high transfection efficiency and stable expression in mouse neuroblastoma (MNA) cells. The intracellular affinity and inhibition of RABV propagation were investigated using RABV-infected MNA cells pretransfected with the scFv-P19 gene. The specific interaction between scFv and RABV-P was confirmed by an immunoprecipitation assay and an indirect immunofluorescence assay showing that these molecules colocalized in the cytoplasm. Measurements of the spread of RABV in a culture well and the virus titer in the supernatant showed that RABV inhibition peaked 3 days after infection, at 98.6% and 99.9% inhibition, respectively. Although the mechanism of RABV inhibition by scFv-P19 is not clear, this scFv-based intracellular immunization could be a candidate for future RABV therapeutic studies if combined with appropriate delivery and application systems.

Xenografts derived from engrafting fresh surgical specimens directly onto immunodeficient mice have enabled the development of more relevant in vivo models for human cancers. Such patient-derived xenograft (PDX) models retain similar morphology, heterogeneities, and molecular signatures with the original cancers. We reported the rapid and efficient establishment of PDXs using super-immunodeficient NOG mice (PDX/NOG models). During the establishment process, a transplantable lymphoproliferative lesion (LPL) that replaces the original tumor cells sometimes occurs. The occurrence of LPL is the most problematic aspect of the establishment of PDX. This phenomenon may arise because of the Epstein-Barr virus due to the severely immunodeficient nature of the animal model; however, the mechanisms underlying LPL have not yet been elucidated in detail. In the present study, we histologically analyzed and tried to prevent the occurrence of LPL in PDX/NOG. Over 100 lines of cancer xenografts were established in our previous studies. Throughout these processes, we also established 10 LPL-PDX lines. Some LPL-PDX lines consist of B cells, T cells, and differentiated plasma cells. These LPL lymphocytes exhibited negligible proliferative potency in vitro. The invasions of LPL in the liver, lung, and pancreas were observed immunohistochemically. NOG mice bearing LPL-PDX (LPL-PDX/NOG) were treated with the anti-graft-versus-host-disease (anti-GVHD) drugs FK506 (1.0 mg/kg/day) and mycophenolate mofetil (MMF, 120 mg/kg/day) for 4 weeks using intraperitoneal injections. The LPL proliferations were significantly suppressed with FK506 (p=0.011) and MMF (p=0.014) compared with the control. The occurrence of LPL during establishment of PDX may be based on the GVHD-like mechanism. This phenomenon can be prevented by using anti-GVHD agents in the severely immunodeficient mice. The LPL-PDX/NOG models are also proposed for the utilization of GVHD mouse models with organic damage, particularly chronic GVHD mouse models. Citation Format: Tsuyoshi Chijiwa, Akira Noguchi, Daisuke Komura, Makoto Katayama, Mizuha Haraguchi, Haruo Hashimoto, Hiroshi Suemizu, Yoshiyasu Nakamura, Daisuke Furukawa, Takayuki Isagawa, Hiroto Katoh, Takashi Moriya, Shumpei Ishikawa, Masato Nakamura, Yohei Miyagi. The prevention of lymphoproliferative lesions arising in patient-derived cancer xenografts by anti-graft-versus-host-disease agents [abstract]. In: Proceedings of the AACR Special Conference: Advances in Modeling Cancer in Mice: Technology, Biology, and Beyond; 2017 Sep 24-27; Orlando, Florida. Philadelphia (PA): AACR; Cancer Res 2018;78(10 Suppl):Abstract nr B13.

Graph theoretical approach is emerging as a powerful tool for analyzing brain functional network. To compare functional electroencephalography (EEG) network between inphase and anti-phase of bimanual hand movement, 10 healthy subjects were carried out experiments. In the clustering coefficient (CC), the alpha band of dominant motor area (C3) and the beta band of non-dominant motor area (C4) during anti-phase movement were higher than those of in-phase movement. In the betweenness centrality (BC), the alpha band of C3 during anti-phase movement was lower than those of in-phase movement. On the other hand, the gamma band of C4 during anti-phase movement was higher than those of in-phase movement. Additional analysis revealed that patient with stroke in the left middle cerebral artery infraction tended to low CC both movement and low BC during anti-phase movement. Our findings suggest that task-specific functional network alterations reflect on these graph theoretical parameters. In conclusion, the graph theoretical analysis applied to EEG network during bimanual hand movement has a potential to evaluate the brain network function.

Recently, development of high technology has been required for the formation of uniform thin film in manufacturing processes of semiconductor as the precision instruments become more sophisticated. A method called spin coating is often used for spreading photoresist on a wafer surface and drying photoresist film. In spin coating process, photoresist is uniformly spread on the wafer surface by centrifugal force caused by rotating wafer. However, it is a serious concern that streaky lines, which are caused by spiral vortices, appear on the wafer surface and prevent the formation of uniform film in the case of high rotating speed. On the other hand, in the case of low rotating speed, a small hump of the film is formed near the wafer edge. The main purpose of this study is to make clear the drying characteristics of the flowing liquid film on the rotating wafer. Temperature distribution of the flowing liquid film is captured by an infrared thermal video camera and radial gradient of the film temperature is introduced in order to evaluate the drying characteristic of the flowing film under steady state condition. Effects of the flow rate of the liquid film on the film temperature are investigated. The film temperature gradually decreases in the radial direction in all cases. At low rotating speed, the radial gradient of the film temperature is almost constant widely. On the other hand, at high rotating speed, the radial gradient of the film temperature takes a certain maximum value. It is found that the location of the gradient peak corresponds with the transition region of the air boundary layer, in which spiral vortices swirl, and shifts to the inner side of the disk with an increase of the liquid flow rate.

We report the case of a 68-year-old male non-smoker suffering from shortness of breath for 1 year. He was referred to our institution for a definitive diagnosis. Both chest X-ray and computed tomography scans showed bilateral hydrothorax with no pleural thickening. We performed flexi-rigid thoracoscopic pleural biopsy on the right side with a single port under local anaesthesia. Multiple white nodules were seen in parietal and visceral pleura, and these nodules were small and flat under white light. Narrow-band imaging demonstrated pathognostic findings on parietal pleura. Irregular dilative vessels were seen around these nodules. Subsequently, we performed parietal pleural biopsy for these nodules. Pathological examination with haematoxylin and eosin staining revealed eosinophilic matrix material depositions present in the pleural parenchyma and the vessel wall. These depositions were positive for Congo red stain and showed apple-green birefringence under polarized light. These findings were compatible with pleural amyloidosis.

Nipah virus (NiV) is a new zoonotic paramyxovirus that emerged in 1998 and is now classified in the genus Henipavirus along with the closely related Hendra virus (HeV). NiV is highly pathogenic in several vertebrate species including humans, and the lack of available vaccines or specific treatment restricts it to biosafety level 4 (BSL4) containment. A serum neutralization test was developed for measuring NiV neutralizing antibodies under BSL2 conditions using a recombinant vesicular stomatitis virus (VSV) expressing green fluorescent protein (GFP) and bearing the F and G proteins of NiV (VSV–NiV–GFP). The neutralization titers were obtained by counting GFP-expressing cells or by measuring fluorescence. The performance of this new assay was compared against the conventional test using live NiV with panels of sera from several mammalian species, including sera from NiV outbreaks, experimental infections, as well as HeV-specific sera. The results obtained with the VSV–NiV–GFP based test correlated with those obtained using live NiV. Using a 50% reduction in VSV–NiV–GFP infected cells as the cut-off for neutralization, this new assay demonstrated its potential as an effective tool for detecting NiV neutralizing antibodies under BSL2 containment with greater speed, sensitivity and safety as compared to the conventional NiV serum neutralization test.

Fixed rabies viruses (CVS-11 strain) were inoculated intramuscularly to C57BL/6J mice, and the pathomorphological changes of the spinal cord including dorsal root spinal ganglion cells were investigated. At 4 days postinoculation (PI), viral antigens were first detected in the spinal neurons and dorsal root spinal ganglion cells without producing morphological changes. At 5 days PI, mild infiltration of lymphocytes was observed around the central canal, small blood vessels and leptomeninges. Cells positive to anti-Iba1 and anti-GFAP antibodies increased significantly from 3 to 5 days PI, respectively. Microglia changed their morphological forms to be ramified or amoeboid, and astroglia extended their cytoplasm from the leptomeninges to the parenchyma. At 7 days PI, apoptotic cells were found in the spinal cord and dorsal root spinal ganglion using TUNEL. We confirmed that most of T lymphocytes and a minority of microglial cells underwent apoptosis, using a combination of TUNEL and immunostaining with antibodies to viral phosphoprotein, CD3, Iba1 and GFAP. On the other hand, astroglial cells and virus-infected nerve cells were negative against TUNEL and cleaved caspase-3 antibody. These findings indicate that T lymphocytes and microglial cells died by apoptosis, whereas virus-infected nerve cells died by necrosis. This was accompanied by increased numbers and morphological changes of glial cells associated with the pathogenesis of CVS-11 in the C57BL/6J mouse.

Although the neural cell adhesion molecule (NCAM) -140 and -180 have been shown to serve as a receptor for rabies virus (RV), it was not known whether the other major isoform of NCAM, GPI-anchored NCAM-120 functions as RV receptor. In this study, we have established HEp-2 cells stably expressing NCAM-120 or NCAM-140, and their susceptibilities to RV infection were compared. The results demonstrated that NCAM-120 served as virus attachment protein; however, the cells expressing NCAM-120 did not support efficient RV replication. Furthermore, the level of IFN-ss mRNA was apparently elevated in NCAM-120 expressing cells but not in NCAM-140 expressing cells, suggesting that GPI-anchored NCAM-120 suppressed RV replication via induction of IFN-ss even though NCAM-120 was able to promote virus penetration into the cells.

With the increase in network traffic due to the increasing use of the Internet, the number of network appliances such as routers and links, as well as power consumption for those appliances, is increasing. Meanwhile, considering the demand for reduction in power consumption for networks, it is desired to halt network appliances that have little traffic. However, when a network appliance is halted, there is a possibility that traffic will concentrate at a specific network appliance that is operating, which may result in congestion. Furthermore, since there are variations in the amount of traffic on real networks, it is necessary to perform switching for booting and halting routers, links, etc. Therefore, control is needed such that appliances will be halted as much as possible while avoiding congestion in view of variations in the amount of traffic. Accordingly, in the research reported in this paper, we designed a method of controlling halting and booting of links and nodes in a situation where the amount of traffic varied, and we evaluated the effectiveness of the proposed control method through numerical experiments.

In this study, G proteins of the rabies virus (RABV) Kyoto strain were detected in the cytoplasm but not distributed at the cell membrane of mouse neuroblastoma (MNA) cells. G proteins of CVS-26 were detected in both the cell membrane and perinuclear space of MNA cells. We found that N-glycosylation of street RABV G protein by the insertion of the sequon Asn(204) induced the transfer of RABV G proteins to the cell surface membrane. Fixed RABV budding from the plasma membrane has been found to depend not only on G protein but also on other structural proteins such as M protein. However, the differing N-glycosylation of G protein could be associated with the distinct budding and antigenic features of RABV in street and fixed viruses. Our study of the association of N-glycan of G protein at Asn(204) with the transport of RABV G protein to the cell surface membrane contributes to the understanding of the evolution of fixed virus from street virus, which in turn would help for determine the mechanism underlying RABV budding and enhanced host immune responses.

The "extraction method" that is commonly used to evaluate the availability of nutrients in soil often involves complex, problematic and simultaneous assays of multiple components and comparisons of analytical values. In a previous paper using pot experiments of plants, we confirmed the possibility of using nutrient concentrations in the xylem sap of Lwu cylindricu Roem. seedlings as an indicator of nutrient availability in soil and suggested that the "xylem sap method," might represent a solution of the above-mentioned problems and inaccuracies characteristic of chemical extraction. To determine whether a method enables to evaluate available nutrient contents in soil, the criterion should probably be that the relationship between the values obtained by that method and the amounts of nutrients absorbed by plants does not vary depending on the types of soil. In this regard, we compared the conventional extraction method with our proposed xylem sap method. In the present paper, we attempted to change the cation exchange capacity (CEC) in soil by the addition of zeolite and/or humic acid to soil to change the nutrient availability without changing the total quantity of nutrients in soil in the pots. Our results showed that the amounts of nutrients absorbed by plants changed under these treatments. However, in some cases, changes in the amounts of phosphorus, calcium, magnesium, and zinc obtained via different extractants did not correspond to the changes in the amounts of nutrients absorbed by the plants. Thus, the amounts of nutrients absorbed by the plants were compared with nutrient concentrations in the xylem sap and the amounts of nutrients inferred by each extractant to examine their relationship. A highly positive correlation was consistently found between the amounts of nutrients absorbed by the plants and their concentrations in the xylem sap for every nutrient. By contrast, there was a lower correlation for the extraction method with some extractants. For phosphorus in particular, no close correlation was obtained between the amounts of nutrients absorbed by the plants and the amounts extracted, regardless of the extractants. In the present study, because we attempted to change the nutrient availability by artificially controlling the soil CEC value, without changing the total amount of nutrients present in the culture medium, the changes in nutrient availability were not conspicuous. Thus, it is possible that the extraction method itself generates large errors for such small changes. Therefore, we concluded that the xylem sap method is superior to the extraction method, as an indicator of nutrient availability in soil, at least under the conditions of the present experiment.

To examine the availability of nutrients in soil, chemical extraction methods are commonly used. However, such methods require different extractant solutions depending on the types of soil and nutrients being assayed, making it difficult to assay multiple components simultaneously and compare analytical values. We proposed an alternative method, consisting of the collection of xylem sap exuded from plant stems by employing plant roots as a tool for evaluating the levels of available nutrients in soil. We designated this method as the "xylem sap method." We tested the effectiveness of this method by growing LWu cylindrica Roem. seedlings for pre-determined periods of time in pots containing culture medium, removing the aerial parts, and collecting the xylem sap from the cut ends. In a sand culture medium, in which all the nutrients provided were considered to be present in an instantly available form, the concentrations of the nutrients in the xylem sap were found to be proportional to those of the nutrients in the culture solution supplied to the medium, confirming that this technique gives a good index of nutrient availability in the rhizosphere. Furthermore, the contents of the nutrients in the xylem sap were proportional to those of the nutrients accumulated in shoots during the growth period. This shows that, at the seedling stage, nutrient uptake by the aerial parts of L. cylindrica during the growth period exactly mirrors the contents in the xylem sap. It was further conhed that the nutrient contents in the xylem sap were similarly representative of the nutritional status of the plant. However, in a soilbased culture medium, the increase in the total amount of phosphorus and micronutrients in the medium was not reflected in the xylem sap. This was in agreement with existing findings that concentrations of certain specific nutrients are maintained at a fairly steady level in the plant, irrespective of their availability in soluble form in soil. Thus, we concluded that changes in most nutrient concentrations in the xylem sap corresponded to changes in the behavior and availability of nutrients in soil and that the xylem sap method could be used as an accurate indicator of the availability of most nutrients in soil.