Dry eye disease is an ocular disease in which the stability of tear fluid decreases, causing ocular discomfort and abnormal visual function as well as damage to the ocular surface. It has been reported that specific types of food ingredients can promote lachrymal secretion, which is expected to prevent and improve dry eye. Here, we evaluated the effects of a heat-killed form of Fructobacillus fructosus FMO-85, a species of fructophilic lactic acid bacteria (FLAB) that is derived from the digestive tract of honeybees, on lacrimal fluid secretion using a stress-induced dry eye mouse model. Male C57BL/6J mice were fed a 3% FLAB-mixed diet 3 weeks before stress loading. We observed that the tear fluid volume was decreased after stress loading, which was significantly improved by FLAB treatment after 7 and 11 days. Mechanistically, the mRNA levels of brain-derived neurotrophic factor (Bdnf), one of the important growth factors involved in lacrimal fluid secretion, isoform-2 and -6 were increased in the hippocampus of FLAB-treated mice. Furthermore, the plasma levels of an anti-inflammatory cytokine, interleukin-10 (IL-10), increased in the FLAB-treated mice. These results suggested that the ingredients contained in dried FLAB increase tear fluid volume by affecting the lacrimal secretion mechanism and the production of inhibitory cytokines. In conclusion, the decrease in tear fluid volume after stress loading was suppressed by FLAB intake. These results indicated that FLAB supplementation may be a useful strategy for the prevention and treatment of dry eye.

The Committee on Sick House Syndrome: Indoor Air Pollution, established by the Ministry of Health, Labour and Welfare of Japan, is reviewing indoor air quality guidelines. A comprehensive exposure assessment is essential for pollutants with revised guideline values or newly developed candidate pollutants, necessitating the development of standardized test methods for an accurate evaluation. However, the available test methods that have been provided as a standard test method (measurement manual) were introduced over 20 years ago. Its applicability to pollutants for which guideline values have been established since then had not been examined. Therefore, we established a test method for six compounds based on the current guideline values and three candidate compounds that underwent initial risk assessment. This method considered the new guideline values established after 2001 using solid-phase adsorption-thermal desorption-gas chromatography/mass spectrometry, as indicated in the measurement manual for volatile organic compounds. This method was validated at four institutions using samples at approximately 1/10^th the concentration of the current, revised, and newly proposed guideline values, as of 2017. Results revealed that the average recovery of the four laboratories ranged from 84.2 to 95.6%, the repeatability ranged from 0.43 to 16%, which was <20%, thereby effectively achieving the target evaluation criteria. Therefore, this method could be presented as a standard test method for nine volatile organic compounds.

To avoid harm caused by falsified medicines, we aimed to devise a method to identify falsified medicines in Japan using visual observation among medicines obtained from personal import agency websites, which are the main conduits through which falsified medicines are obtained. We recorded details regarding the information provided on personal import agency websites used to purchase medicines, the outer package received, the customs declaration description, and the product appearance for 212 samples of medicines obtained through personal import via the Internet. We investigated the relationship between each observed item and the rate of falsified medicines. We developed a classification and prediction model to identify falsified medicines using items that could be visually observed. The results showed that the rate of falsified medicines was significantly higher for websites that did not contain identifying information such as the name and address of the contact or import agency, as well as for products that did not contain the name and address of the manufacturer, indicating that these items may be useful in the identification of falsified medicines. In the prediction model constructed, we extracted features such as the country of dispatch and address of the import agency, and a prediction model was created to identify falsified medicines and websites selling these medicines. Careful observation of the identified features and use of our prediction model will help to prevent harm owing to the use of falsified medicines.

Bisulfite modification of cytosine residues is a widely used method for analyzing genomic DNA methylation. Despite its robustness, degradation of DNA during modification has hampered the application to the analysis of small amounts of DNA. We show that the addition of large volume bisulfite-treated heterogeneous DNA promotes the amplification of the targeted region in bisulfite-treated DNA by PCR. The addition of untreated DNA did not promote the amplification. The addition of large volume bisulfite-treated heterogeneous DNA neither promoted the amplification when untreated DNA was used as template. PCR products were detected when a ten thousandth of aliquot of bisulfite-treated 1 μg of human genomic DNA (0.1 ng) was used as template. This figure is equivalent to that of human genomic DNA derived from as little as 15 cells. This procedure permits the analysis of genomic DNA methylation when only limited numbers of cells are available.

Semaphorin family belongs to secreted or membrane anchored proteins. Recent studies have demonstrated the involvement of semaphorins and their receptors in cancer biology, but it remains unclear how each semaphorin molecule regulates tumorigenesis. Previously we reported that Semaphorin 3A (Sema3A) and its receptor Plexin A1 (PlxnA1) regulate the malignant phenotypes of mouse-derived lewis lung cancer (LLC) cells constitutively expressing GFP (LLC-GFP). Here we show that Semaphorin 6B (Sema6b) serves as one of the oncogenic semaphorin molecules in lung cancer using LLC-GFP. Sema3a or Plxna1 knockdown downregulated Sema6b, and their suppressive effect on proliferation was significantly recovered by recombinant SEMA6B (rSEMA6B) treatment. Sema6b knockdown suppressed the proliferation and tumorigenicity of LLC-GFP. Interestingly, the self-renewal capacity of LLC-GFP derived cancer stem-like cells (LLC-GFPstem) was completely lost by Sema6b knockdown. These results demonstrate that Sema6B would be the novel therapeutic target of lung cancer.

Iron is an essential nutrient for bacterial survival. Vibrio alginolyticus is a pathogenic Vibrio species that produces vibrioferrin, a cognate siderophore for efficient iron acquisition in iron-limited environments. Many bacteria have developed mechanisms to utilize xenosiderophores produced by other microorganisms, in addition to using self-produced siderophores for iron acquisition. In this study, we found through a homology search using the whole genome sequence of V. alginolyticus NBRC 15630 that this bacterium has genes similar to those involved in the utilization of hydroxamate-based xenosiderophores, desferri-ferrichrome (DFC), desferrioxamine B (DFOB), and aerobactin (AERO), possessed by V. parahaemolyticus, V. cholerae, and V. vulnificus. In growth assays using an iron-limiting medium supplemented with each xenosiderophore, it was found that the N646_3157 (fhuA1), N646_0489 (fhuA2), N646_0777 (desA), and N646_4356 (iutA) are outer membrane receptor genes involved in the utilization of DFC, DFOB, and AERO and N646_3158-3160 (fhuC1D1B1), and N646_0486-0488 (fhuC2D2B2) are ATP-binding cassette transporter genes for both DFC and DFOB. Additionally, we demonstrated by reverse transcriptase-quantitative PCR and electrophoretic mobility shift assay that the fhuC2D2B2A2 genes, which were newly identified in pathogenic Vibrio species, are an operon whose expression is probably regulated by Fur in response to iron availability.

OSW-1, a promising compound that is toxic to diverse tumor cell lines, is a saponin from Ornithogalum saundersia. In this study, we analyzed the stress responses induced by OSW-1 using the human colon cancer cell line HT-29 cells and compared it with the commonly used ER and Golgi stress inducers brefeldin A (BFA), thapsigargin (Tg), and tunicamycin (Tm). OSW-1 induced few ER stress-related factors, but there was an increase in expression of TFE3 protein, one of the Golgi stress response factors. A shift in the molecular weight of TFE3 was also found, likely attributable to dephosphorylation. Conversely, the impact of OSW-1 on the expression of the TFEB protein, another member of the MiTF/TFE family, was minimal. Cleavage of CREB3, another Golgi stress sensor, was apparently induced only by BFA. LC3-II and p62, autophagy-related factors, were increased in all drug treatments. Unexpectedly, OSBP protein levels, one of the targets of OSW-1, were increased by not only three reagents but also OSW-1. Taken together, OSW-1 treatment of HT-29 cells induced atypical Golgi stress that strongly activated the TFE3 pathway and did not involve the CREB3 pathway or the ER stress response. Although OSW-1 was also found to affect the autophagy system, it was suggested that the effects of OSW-1 may not be mediated by OSBP depletion. These findings will contribute to the development of OSW-1-based cancer therapies and to our understanding of Golgi stress responses.