The A2'N mutation of the RDL gamma-aminobutyric acid receptor conferring fipronil resistance in Laodelphax striatellus (Hemiptera: Delphacidae).

The planthopper Laodelphax striatellus (Fallén) (Hemiptera: Delphacidae) is a serious insect pest of rice, Oryza sativa L., and has developed resistance to fipronil in Japan. Sequence analysis of L. striatellus RDL gamma-aminobutyric acid (GABA) receptor subunit (LS-RDL) genes from a fipronil-resistant population and a fipronil-susceptible strain identified the A2'N mutation (index number for M2 membrane-spanning region), that was previously implicated in fipronil resistance in the planthopper Sogatella furcifera (Horváth) (Hemiptera: Delphacidae). Nineteen of 21 fipronil-resistant L. striatellus individuals were genotyped as heterozygous for the A2'N mutation, suggesting that this mutation is associated with fipronil resistance and that most fipronil-resistant L. striatellus express wild-type and A2'N mutant LS-RDL simultaneously. To confirm the role of the A2'N mutation of LS-RDL, Drosophila Mel-2 cells were transfected with wild-type and A2'N mutant LS-RDL genes, either individually or together. A membrane potential assay showed that fipronil had no inhibitory effect at 10 microM on cells transfected with the A2'N mutant LS-RDL gene with or without the wild-type LS-RDL gene. By contrast, the IC50 value of fipronil for wild-type LS-RDL homomers was 14 nM. These results suggest that the A2'N mutation of the RDL GABA receptor subunit confers fipronil resistance in L. striatellus as well as S. furcifera.

Evaluation of antimicrobial use in dairy cattle, beef cattle and broilers in Japan using dosage-based indicators.

The sales amount of antimicrobials intended for use in dairy cattle, beef cattle and broilers from 2008 to 2019 was evaluated for each antimicrobial class and administration route using dosage-based indicators. Our results revealed that the antimicrobial sales amount sold for use in dairy cattle in 2019 in terms of total weight of active ingredient, the number of defined daily doses (DDDs) (theoretical amount of biomass subjected to antimicrobial treatment in a year) and the number of treatment days (TDs) (theoretical number of days of treatment that an animal is subjected to in a year) calculated using Japanese DDD values (DDDjp values) was 36,751 kg, 8,261,848,000 kg·days and 15.5 days, respectively. Likewise, the antimicrobial sales amount sold for use in beef cattle and broilers in 2019 in terms of these metrics was 33,403 kg, 3,928,248,000 kg·days and 3.61 days, and 69,773 kg, 2,947,848,000 kg·days and 10.66 days, respectively. There was a considerable difference between the number of DDDs calculated using DDDjp values and that calculated using European DDD values (DDDvet values) for antimicrobial products sold for use in dairy and beef cattle. Our results also revealed that the sales amount of some antimicrobials, such as cephalosporins and quinolones represented larger proportions when calculated using dosage-based indicators than when calculated using the weight of active ingredient. Considering that Japanese veterinarians and farmers are more likely to conform to the Japanese dosage recommendations rather than the European ones, these results indicate the need for using dosage-based metrics, in particular metrics based on Japanese dosages rather than European dosages.

Establishing defined daily doses (DDDs) for antimicrobial agents used in pigs, cattle and poultry in Japan and comparing them with European DDD values.

Monitoring of antimicrobial use is essential in the management of the development and selection of antimicrobial resistance. A variety of indicators has become available to monitor antimicrobial use in human and animal medicine. One of them is an indicator based on defined daily dose (DDD). By using the number of DDDs administered and normalising it by the population at risk of being treated over a defined period, one can estimate the number of treatment days with antimicrobial agents in a population. For veterinary medicine, the European Medicines Agency (EMA) has published the European values of DDD (DDDvet) for food-producing animals. In this study, we defined Japanese defined daily doses for antimicrobial agents (DDDjp) using DDD values that we previously assigned for antimicrobial products approved for use in pigs, cattle and poultry in Japan and compared them with DDDvet values. For the comparison, the quotient of Japanese and European values (QDDD) was calculated and the effect of the administration route and the number of active substances contained in the preparation was investigated. A total of 59 DDDjp values were defined for 43 antimicrobial agents using the data of 276 products approved for use in pigs. Likewise, a total of 55 DDDjp values were defined for 32 antimicrobial agents using the data of 196 products for use in cattle, and a total of 27 DDDjps values were defined for 25 antimicrobial agents using the data of 131 products approved for use in poultry. A comparison was made for 42, 28 and 17 pairs of DDDjp and DDDvet values for antimicrobial agents used for pigs, cattle and poultry respectively. The comparison showed median QDDD value of 0.61 and 0.66 for antimicrobial agents used for pigs and cattle respectively (p<0.01), indicating that the Japanese daily doses are significantly lower than the corresponding EMA values in these species. For the antimicrobial agents used for poultry, no significant difference was observed between DDDjp and DDDvet values with a median QDDD value of 1.15. The difference between DDDvet and DDDjp values and absence of DDDvet values for some antimicrobial agents marketed in Japan indicate that DDDjp rather than DDDvet should be used as the basis for the calculation of antimicrobial use monitoring in farm animals in Japan.

Evaluation of the antimicrobial use in pigs in Japan using dosage-based indicators.

The use of antimicrobial agents in food-producing animals may lead to the emergence and spread of antimicrobial resistance in bacteria of animal origin. The use of antimicrobial agents in pigs in 2018 in Japan was evaluated in terms of the weight of active ingredient and number of defined daily doses (DDD), using annual sales data of veterinary antimicrobials sold for use in pigs. In addition, the use of antimicrobial agents in the Japanese pig sector in 2008 to 2017 was evaluated to determine whether or not there were any differences in temporal change pattern by use of different metrics. In 2018, 447 metric tons of active ingredient, corresponding to 77,379 × 106 kg-days (Japanese DDD) and 34,903 × 106 kg-days (European DDD) were sold. The proportion of the sales amount of sulfonamides, trimethoprim and lincosamides to the total sales amount was significantly different depending on the metric used. For most antimicrobial classes, the number of Japanese DDDs was greater than the number of European DDDs. These results indicate that the DDD-based metric, which is more reflective of the selective pressure of antimicrobials, is recommended for use in monitoring the antimicrobial use in pigs in Japan. The differences in the number of Japanese DDDs and European DDDs appear to confirm the need for Japanese DDDs.

Monitoring biodistribution of glycoproteins with modified sugar chains.

Natural human interferon (hIFN)-gamma has mainly biantennary complex-type sugar chains. Previously, we successfully remodeled its sugar chain structure into: (a) highly branched types; or (b) highly sialylated types, by overexpression of: (a) N-acetylglucosaminyltransferase (GnT)-IV and/or GnT-V; or (b) sialyltransferases, in Chinese hamster ovary (CHO) cells. In addition, we prepared asialo hIFN-gammas by treatment with sialidase in vitro. In the present study, we assessed the bioactivity of remodeled hIFN-gamma in terms of antiviral activity, anticellular activity, and biodistribution. Structural changes to the sugar chains did not have a significant influence on the antiviral and anticellular activities of hIFN-gamma, although the attachment of the sugar chain itself affected both activities. However, the biodistribution differed significantly; the number of exposed galactose residues was the major determinant of the specific distribution to the liver and blood clearance rate of hIFN-gamma. This phenomenon was considered to be mediated by the hepatic asialoglycoprotein receptor (ASGP-R), and we showed a linear, not exponential, enhancement of the distribution to the liver with an increase in the number of exposed galactose residues. We also confirmed this tendency using fibroblast growth factor (FGF). Our observation is not the same as the "glycoside cluster effect." We thus provide important information on the character of modified recombinant glycoproteins.

HPLC profile of captopril disulfide that undergoes reversible cis-trans conversion among three isomers.

Peculiarly shaped chromatograms of some compounds that consist of two reversible isomers have been reported. Those of a compound that consists of three reversible isomers are described here. Because disulfide of captopril has two cis-trans convertible bonds, it exists in three forms of cis-cis, cis-trans and trans-trans isomers. The disulfide has produced various HPLC-UV profiles under various conditions, such as three split peaks, three peaks with bridging plateaus and one broad peak. The effect of the column temperature and the eluent flow rate on the profile was investigated and explained by interconversion among three isomers. A profile analysis could provide some kinetic constants of this reaction.

Parietal and frontal object areas underlie perception of object orientation in depth.

Recent studies have shown that the human parietal and frontal cortices are involved in object image perception. We hypothesized that the parietal/frontal object areas play a role in differentiating the orientations (i.e., views) of an object. By using functional magnetic resonance imaging, we compared brain activations while human observers differentiated between two object images in depth-orientation (orientation task) and activations while they differentiated the images in object identity (identity task). The left intraparietal area, right angular gyrus, and right inferior frontal areas were activated more for the orientation task than for the identity task. The occipitotemporal object areas, however, were activated equally for the two tasks. No region showed greater activation for the identity task. These results suggested that the parietal/frontal object areas encode view-dependent visual features and underlie object orientation perception.

Association of metabolic syndrome with urinary albumin excretion, low-grade inflammation, and low glomerular filtration rate among non-diabetic Japanese subjects.

OBJECTIVE: Several studies have reported a significant association of metabolic syndrome with urinary albumin excretion, high-sensitivity C-reactive protein, or chronic kidney disease; however, no study has investigated the association of metabolic syndrome with these 3 factors together in the same individual. Therefore, we conducted the present study to obtain more information on this association. METHODS: We enrolled 712 Japanese subjects without diabetes, macroalbuminuria, or medications, who entered our hospitalized health check-up program (180 women and 532 men; mean age, 53.2 years; mean body mass index, 24.1 kg/m(2)). Metabolic syndrome was diagnosed by 4 major definitions. Low glomerular filtration rate was defined as an estimated glomerular filtration rate of <60 mL/min/1.73 m(2). RESULTS: Both urinary albumin excretion and high-sensitivity C-reactive protein were significantly higher in those with metabolic syndrome than without, and metabolic syndrome was an independent determinant of both. In contrast, estimated glomerular filtration rate and the prevalence of low glomerular filtration rate did not differ significantly between those with and without metabolic syndrome. Among the 5 components of metabolic syndrome and other clinical variables, systolic blood pressure was an independent determinant of urinary albumin excretion; the 5 components and low-density lipoprotein cholesterol were all independent determinants of high-sensitivity C-reactive protein; systolic blood pressure was an independent determinant of low glomerular filtration rate. CONCLUSION: Metabolic syndrome is associated with vascular dysfunction and low-grade inflammation and the latter association is strong, whereas the association of metabolic syndrome with low glomerular filtration rate may be less apparent among those without diabetes, macroalbuminuria, and medications.