Prunin Laurate Derived from Natural Substances Shows Antibacterial Activity against the Periodontal Pathogen Porphyromonas gingivalis.

Periodontal disease is an inflammatory disease caused by infection with periodontopathogenic bacteria. Oral care is essential to prevent and control periodontal disease, which affects oral and systemic health. However, many oral hygiene products currently on the market were developed as disinfectants, and their intense irritation makes their use difficult for young children and older people. This study investigated the antibacterial effects of prunin laurate (Pru-C12) and its analogs on periodontopathogenic bacteria, Porphyromonas gingivalis (P. gingivalis). Pru-C12 and its analogs inhibited in vitro bacterial growth at more than 10 µM and biofilm formation at 50 µM. Among its analogs, only Pru-C12 showed no cytotoxicity at 100 µM. Three of the most potent inhibitors also inhibited the formation of biofilms. Furthermore, Pru-C12 inhibited alveolar bone resorption in a mouse experimental periodontitis model by P. gingivalis infection. These findings may be helpful in the development of oral hygiene products for the prevention and control of periodontal disease and related disorders.

Homogalacturonan and xylogalacturonan region specificity of self-cloning vector-expressed pectin methylesterases (AoPME1-3) in Aspergillus oryzae.

Aspergillus oryzae is a safe microorganism that is commonly used in food production. We constructed a self-cloning vector capable of high expression in A. oryzae. Using the vector, three putative pectin methylesterase (PME) genes belonging to Carbohydrate Esterase family 8 derived from A. oryzae were expressed, and several characteristics of the gene products were examined. The effects of temperature and pH on the three enzymes (AoPME1, 2, and 3) were similar, with optimal reaction temperatures of 50 - 60 °C and optimal reaction pH range of 5 - 6. The specific activities of AoPME1, 2, and 3 for apple pectin were significantly different (34, 7,601, and 2 U/mg, respectively). When the substrate specificity was examined, AoPME1 showed high activity towards pectin derived from soybean and pea. Although AoPME2 showed little activity towards these pectins, it showed very high activity towards apple- and citrus-derived pectins. AoPME3 showed low specific activity towards all substrates tested. Sugar composition analysis revealed that apple- and citrus-derived pectins were rich in homogalacturonan, while soybean- and pea-derived pectins were rich in xylogalacturonan. When pea pectin was treated with endo-polygalacturonase or endo-xylogalacturonase in the presence of each PME, specific synergistic actions were observed (endo-polygalacturonase with AoPME1 or AoPME2 and endo-xylogalacturonase with AoPME1 or AoPME3). Thus, AoPME1 and AoPME3 hydrolyzed the methoxy group in xylogalacturonan. This is the first report of this activity in microbial enzymes. Our findings on the substrate specificity of PMEs should lead to the determination of the distribution of methoxy groups in pectin and the development of new applications in the field of food manufacturing.

Exopolysaccharides from a Scandinavian fermented milk viili increase butyric acid and Muribaculum members in the mouse gut.

Starter culture of viili contains lactic acid bacteria belonging to Lactococcus lactis. These bacteria secrete large polysaccharides (EPSs) into milk, resulting in a ropy texture of viili. In mouse experiments, a large dose of EPS (5-140 mg/day) has been shown to alleviate severity of artificially induced illness through modulation of the gut microbiota. The present study investigated whether supplementary amounts of EPS affects the gut microbiota of normal mouse. EPS with high glucosamine content (VEPS) was isolated from home-made viili. C57BL/6J male mice fed ordinary diet took 49 ± 1 µg VEPS/day for 28 days by drinking ad libitum tap water containing 8 µg/mL VEPS. The relative abundance of Muribaculum increased significantly by VEPS supplementation. The relative abundance of fecal butyric acid decreased in control mice, and VEPS prevented this decrease. These findings indicated that the gut microbiota can be modulated by a small dose of VEPS.

Inter-agency collaboration factors affecting multidisciplinary workers' ability to identify child maltreatment.

OBJECTIVES: This study aimed to clarify the factors of successful inter-agency collaboration that affect multidisciplinary workers' abilities to identify child maltreatment. A questionnaire-based survey was conducted; the contents of the questionnaire included the Collaboration Evaluation Scale we developed and the workers' abilities to identify child maltreatment. In total, 277 individuals from various agencies in Japan participated in this study. To examine the factors of successful inter-agency collaboration affecting workers' awareness of child maltreatment, we used hierarchical multiple regression analysis. RESULTS: The analysis showed the positive effect of "commitment with loyalty" on the workers' awareness of child maltreatment-related information in all fields (ß = .18-.31, p < .05), the effect of "strong leadership" on information about maltreated children and the home environment (ß = .18, p < .05; ß = .16, p < .05, respectively), and the effect of "resources" on the information about mothers' information during pregnancy and of fathers' feelings towards their children during the perinatal period (ß = .17, p < .05; ß = .22, p < .01, respectively). In conclusion, commitment with loyalty, strong leadership, and resources are factors of successful inter-agency collaboration that affects the ability of multidisciplinary workers to recognize signs of child maltreatment.

Identification and characterization of ferulic acid esterase from Penicillium chrysogenum 31B: de-esterification of ferulic acid decorated with l-arabinofuranoses and d-galactopyranoses in sugar beet pectin.

We previously described the fungus Penicillium chrysogenum 31B, which has high performance to produce the ferulic acid esterase (FAE) for de-esterifying ferulic acids (FAs) from sugar beet pulp. However, the characteristics of this fungus have not yet been determined. Therefore, in this study, we evaluated the molecular characteristics and natural substrate specificity of the Pcfae1 gene from Penicillium chrysogenum and examined its synergistic effects on sugar beet pectin. The Pcfae1 gene was cloned and overexpressed in Pichia pastoris KM71H, and the recombinant enzyme, named PcFAE1, was characterized. The 505 amino acids of PcFAE1 possessed a GCSTG motif (Gly164 to Gly168), characteristic of the serine esterase family. By comparing the amino acid sequence of PcFAE1 with that of the FAE (AoFaeB) of Aspergillus oryzae, Ser166, Asp379, and His419 were identified as the catalytic triad. PcFAE1 was purified through two steps using anion-exchange column chromatography. Its molecular mass without the signal peptide was 75 kDa. Maximum PcFAE1 activity was achieved at pH 6.0-7.0 and 50 °C. The enzyme was stable up to 37 °C and at a pH range of 3-8. PcFAE1 activity was only inhibited by Hg2+, and the enzyme had activity toward methyl FA, methyl caffeic acid, and methyl p-coumaric acid, with specific activities of 6.97, 4.65, and 9.32 U/mg, respectively, but not on methyl sinapinic acid. These results indicated that PcFAE1 acted similar to FaeB type according the Crepin classification. PcFAE1 de-esterified O-[6-O-feruloyl-ß-d-galactopyranosyl-(1→4)]-d-galactopyranose, O-[2-O-feruloyl-α-l-arabinofuranosyl-(1→5)]-l-arabinofuranose, and O-[5-O-feruloyl-α-l-arabinofuranosyl-(1→3)]-O-ß-d-xylopyranosyl-(1→4)-d-xylopyranose, indicating that the enzyme could de-esterify FAs decorated with both ß-d-galactopyranosidic and α-l-arabinofuranosidic residues in pectin and xylan. PcFAE1 acted in synergy with endo-α-1,5-arabinanase and α-l-arabinofuranosidase, which releases FA linked to arabinan, to digest the sugar beet pectin. Moreover, when PcFAE1 was allowed to act on sugar beet pectin together with Driselase, approximately 90% of total FA in the substrate was released. Therefore, PcFAE1 may be an interesting candidate for hydrolysis of lignocellulosic materials and could have applications as a tool for production of FA from natural substrates.

Maintenance electroconvulsive therapy (ECT) for treatment-resistant disorganized schizophrenia.

Patients who have not responded to recommended antipsychotic medications should be considered for electroconvulsive therapy (ECT). However, there has been controversy about the standardized methods of continuation and maintenance ECT in the management of treatment-resistant schizophrenia. We describe a patient with a serious case of disorganized schizophrenia who had not responded well with any typical and atypical antipsychotic drug for seven years, but responded remarkably to acute ECT. Continuation ECT was necessary to sustain a positive therapeutic response. The patient showed dramatic improvement from 70 to 20 in the 18-item Brief Psychiatric Rating Scale (BPRS) score (71% reduction) after acute ECT and continuation ECT. Using maintenance ECT, she was able to live in the custody of her parents after 7-years hospitalization. This case report suggests that continuation and maintenance ECT benefits patients with serious cases of refractory schizophrenia.