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FlhDC is a heterohexameric complex that acts as a master regulator of flagellar biosynthesis genes in numerous bacteria. Previous studies have identified a single flhDC operon encoding this complex. However, we found that two flhDC loci are present throughout Paraburkholderia , and two additional flhC copies are also present in Paraburkholderia unamae. Systematic deletion analysis in P. unamae of the different flhDC copies showed that one of the operons, flhDC1 , plays the predominant role, with deletion of its genes resulting in a severe inhibition of motility and biofilm formation. Expression analysis using promoter- lacZ fusions and real-time quantitative PCR support the primary role of flhDC1 in flagellar gene regulation, with flhDC2 a secondary contributor. Phylogenetic analysis shows the presence of the flhDC1 and flhDC2 operons throughout Paraburkholderia . In contrast, Burkholderia and other bacteria only carry the copy syntenous with flhDC2 . The variations in impact each copy of flhDC has on downstream processes indicate that regulation of FlhDC in P. unamae , and likely other Paraburkholderia species, is regulated at least in part by the presence of multiple copies of these genes. IMPORTANCE Motility is important in the colonization of plant roots by beneficial and pathogenic bacteria, with flagella playing essential roles in host cell adhesion, entrance, and biofilm formation. Flagellar biosynthesis is energetically expensive. Its complex regulation by the FlhDC master regulator is well studied in peritrichous flagella expressing enterics. We report the unique presence throughout Paraburkholderia of multiple copies of flhDC . In P. unamae , the flhDC1 copy showed higher expression and a greater effect on swim motility, flagellar development, and regulation of downstream genes, than the flhDC2 copy that is syntenous to flhDC in Escherichia coli and pathogenic Burkholderia spp. The flhDC genes have evolved differently in these plant-growth-promoting bacteria, giving an additional layer of complexity in gene regulation by FlhDC.

Micro-, small-, and medium-sized enterprises (SMEs) have been considered as key players who can bring innovative medicinal products and/or technologies into the field. However, they may need much regulatory/scientific supports to provide their products, technologies, or services to the market in a timely way. Both the Pharmaceuticals and Medical Devices Agency (PMDA) and the European Medicines Agency (EMA), regulatory authorities for medicinal products in Japan and the EU, respectively, have prepared supportive measures for SMEs from the early phase of product/technology development to the postmarketing phase. With respect to supports for SMEs, both agencies have provided similar SME-specific supportive activities, including routine administrative assistance, consultations about product development strategy from an early phase, as well as specific regulatory/scientific issues and fee incentives. In addition, there is a system to register SME status in the EU, which can be a tool for regulators to know how much potential SME-driven activities have and with whom they should communicate to provide necessary supports. Furthermore, as new technologies and novel products from SMEs are not limited to the region where they are developed, close communication about these topics between the PMDA and the EMA will contribute to advancing patients' access to necessary medicinal products.

Indoor air pollution by volatile organic compounds (VOCs), which may cause a hazardous influence on human being such as sick building (sick house) syndrome, has become a serious problem. In this study, VOCs emitted from nine pieces of home furniture, three sets of dining tables, three sets of chest of drawers and three sofas, were analyzed as potential sources of indoor air pollution by large chamber test method (JIS A 1911). Based on the emission rates of total VOC (TVOC), the impacts on the indoor TVOC was estimated by the sample model with a volume of 20 m3 and ventilation frequency of 0.5 times/h. The estimated TVOC increment values were exceeded the provisional target value for indoor air (400 microg/m3) in three sets of dining tables, one set of chest of drawer and one sofa. The estimated increment of formaldehyde were exceeded the guideline value (100 microg/m3) in one set of dining table, two sets of chest of drawers and one sofa. These results revealed that VOC emissions from furniture may influence significantly indoor air quality. Also, in this study, to establish the alternative method for large chamber test methods, emission rates from representative three parts of furniture unit were evaluated using the small chamber and emission rate from full-sized furniture was predicted. Emission rates of TVOC and formaldehyde predicted by small chamber test were 3-46% and 6-252% of the data obtained using large chamber test, respectively.

The aim of this study was to evaluate aldehydes and other volatile organic compounds (VOCs) emission from furniture, which may cause hazardous influence on human being such as sick building/sick house syndrome. In this study, VOCs emitted from six kinds of wood furniture, including three set of dining tables and three beds, were analyzed by large chamber test method (JIS A 1911). Based on the emission rates of total VOCs (TVOC), the impacts on the indoor TVOC was estimated by the simulation model with volume of 20 m3 and ventilation frequency of 0.5 times/h. The estimated increment of formaldehyde were exceeded the guideline value (100 microg/m3) in one set of dining table and one bed. The estimated TVOC increment values were exceeded the provisional target value for indoor air (400 microg/m3) in two sets of dining tables and two beds. These results revealed that VOC emissions from wood furniture may influence significantly indoor air quality. Also, in this study, to establish the alternative method for large chamber test methods, emission rates from representative five areas of furniture unit were evaluated by passive sampling method using flux sampler and emission rate from full-sized furniture was predicted. Emission rates predicted by flux passive sampler were 10-106% (formaldehyde) and 8-141% (TVOC) of the data measured using large chamber test, respectively.

The Ministry of Health, Labor and Welfare (former MHW) of Japan issued a Directive in 1997 advising restaurants and caterers to freeze portions of both raw food and cooked dishes for at least 2 weeks. This system has been useful for determining vehicle foods at outbreaks. Enumeration of bacteria in samples of stored food provide data about pathogen concentrations in the implicated food. Data on Salmonella concentrations in vehicle foods associated with salmonellosis outbreaks were collected in Japan between 1989 and 1998. The 39 outbreaks that occurred during this period were categorized by the settings where the outbreaks took place, and epidemiological data from each outbreak were summarized. Characteristics of outbreak groups were analyzed and compared. The effect of new food-storage system on determination of bacterial concentration was evaluated. Freezing and nonfreezing conditions prior to microbial examination were compared in the dose-response relationship. Data from outbreaks in which implicated foods had been kept frozen suggested apparent correlation between the Salmonella dose ingested and the disease rate. Combined with results of epidemiological investigation, quantitative data from the ingested pathogen could provide complete dose-response data sets.