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Background: Single-blind, nine case comparative studies were conducted to evaluate salivary fluoride concentrations following toothbrushing using experimental toothpastes containing Surface Pre-Reacted Glass-ionomer (S-PRG) fillers. Preliminary tests were conducted in order to determine the volume of usage as well as the concentrations (wt%) of S-PRG filler. Based on the results given these experiments, we compared the salivary fluoride concentrations following toothbrushing with 0.5 g of 4 different types of toothpastes: 5 wt % S-PRG filler, 1400 ppm F AmF (amine fluoride), 1500 ppm F NaF (sodium fluoride), and MFP (monofluorophosphate) containing toothpaste. Methods: Of the 12 participants, 7 participated in the preliminary study and 8 participated in the main study. All participants brushed their teeth using the scrubbing method for 2 minutes. At first 1.0 and 0.5 g of 20 wt % S-PRG filler toothpastes were used to compare, then followed by 0.5 g of 0 (control), 1, and 5 wt % S-PRG toothpastes, respectively. ThepParticipants spat out once and rinsed with 15 mL of distilled water for 5 seconds. Saliva was collected for 3 minutes each at different time intervals of 0 (baseline), 5, 10, 15, 30, 60, 120, and 180 minutes (min) after the rinsing. Fluoride concentrations were determined using a fluoride-electrode, and the area under the curve (AUC: ppm‧min) of each toothpaste was calculated as the salivary fluoride retention. The main study was then conducted to evaluate the salivary fluoride concentrations as well as the AUC value using 0.5 g of 5 wt % S-PRG filler toothpaste, followed by NaF, MFP, and AmF toothpaste. Results: Since there were no statistical differences between using 1.0 g and 0.5 g of 20 wt % S-PRG toothpastes in salivary fluoride concentrations as well as the AUC value throughout the 180 min measurement, the volume was set as 0.5 g for the following studies. Concentrations of 5 and 20 wt % S-PRG toothpastes retained 0.09 ppm F or more in saliva even after 180 min. No statistical differences were seen in the salivary fluoride concentrations at any time intervals as well as the AUC value between 5 and 20 wt % S-PRG toothpastes. Based on these results, the concentration of 5 wt % S-PRG toothpaste was used for the main comparative study. MFP toothpaste resulted in by far the lowest salivary fluoride concentrations (0.06 ppm F at 180 min) and the AUC value (24.6 ppm‧min), whereas 5 wt % S-PRG toothpaste (0.15 ppm F at 180 min, 92.3 ppm‧min) displayed retention on par with AmF toothpaste which appeared to result in higher values (0.17 ppm F at 180 min, 103 ppm‧min), compared to NaF toothpaste (0.12 ppm F at 180 min, 49.3 ppm‧min). Conclusions: The salivary fluoride concentrations following toothbrushing with 0.5 g of 5 wt % S-PRG filler containing toothpaste showed retention similar to the best performing 1400 ppm F AmF toothpaste even 180 min after toothbrushing.

The spread of methicillin resistance and virulence among staphylococci in the community poses a public health concern. In this study, we investigated the prevalence of

Understanding of the long-term soil fertility trends is the basis for rational and sustainable management of the soil resources in agricultural fields. In tropical Asia, the Green Revolution initia...

The acquisition of drug resistance and virulence by staphylococcal species colonizing humans is a growing public health concern. The present study was conducted to investigate the prevalence, antimicrobial resistance and genetic characteristics of Staphylococcus isolates from the oral cavity and skin (hand) of systemically healthy subjects with dental disease and dental staff in northern Japan. Among a total of 133 subjects (91 patients and 42 staff), 87 coagulase-positive Staphylococcus (83 S. aureus/4 S. argenteus) and 162 coagulase-negative Staphylococcus (CoNS) isolates were recovered from 59 (44.4%) and 95 (71.4%) subjects, respectively. Three oral isolates were methicillin-resistant S. aureus (MRSA) (3.6%, 3/83) that were genotyped as ST8-SCCmec-IVl, ST4775(CC1)-SCCmec-IVa and ST6562(CC8)-SCCmec-IVa. Remarkably, the ST6562 isolate harbored PVL genes on ΦSa2usa and type I ACME (arginine catabolic mobile element). Four methicillin-susceptible isolates were identified as S. argenteus belonging to ST1223 and ST2250, which harbored enterotoxin genes egc-2 and sey, respectively. Among the fourteen CoNS species identified, methicillin-resistant (MR) isolates were detected in five species (11 isolates, 13.3% of CoNS), with S. saprophyticus and S. haemolyticus being the most common. ACME was prevalent in only S. epidermidis and S. capitis. These findings indicated the potential distribution of USA300 clone-like MRSA, toxigenic S. argenteus and MR-CoNS in the oral cavity of dental patients.

Staphylococcus argenteus, a novel staphylococcal species independent of S. aureus, causes a wide spectrum of infectious diseases. As detection of this species from humans and animals has been increasingly reported worldwide, its growing virulence and drug resistance via external genetic determinants has become concerning. In this study, the prevalence and genetic characteristics of virulence factors and drug resistance determinants were investigated for 82 S. argenteus clinical isolates in Hokkaido, Japan, for a one-year period starting in August 2019. These S. argenteus isolates corresponded to 0.66% of the total number of S. aureus isolates collected in the same period. The most prevalent genotype was sequence type (ST) 2250 and staphylocoagulase (coa) genotype XId (45.1%, n = 37), followed by ST1223-coa XV (30.5%, n = 25) and ST2198-coa XIV (24.4%, n = 20). Panton-Valentine leukocidin genes (lukS-PV-lukF-PV) were identified in a single ST2250 isolate. Only ST1223 isolates had the enterotoxin gene cluster (egc-2), seb, and selw (detection rate, 100%, 60%, and 84%, respectively), while sec, sey, sel26-sel27, tst-1 were only detected in ST2250 isolates (detection rate, 10.8%, 100%, 67.6%, and 10.8%, respectively). ST2198 isolates harbored selx at a significantly higher rate (60%) than isolates of other STs. Although most of S. argenteus isolates were susceptible to antimicrobials examined, ST2198 showed higher resistance rates to penicillin, macrolides, and aminoglycosides than other STs, and it harbored various resistance genes such as blaZ, erm(C), msr(A), lnuA, and aac(6′)-Ie-aph(2″)-Ia. Only one ST2250 isolate possessed SCCmec-IVc, showing resistance to oxacillin. blaZ was the most prevalent determinant of resistance in the three STs and belonged to two plasmid groups and a chromosomal group, suggesting its diverse origin. lnu(A) in ST2198 isolates was assigned to a major cluster with various staphylococcal species. The present study indicates that the prevalence of virulence factors and drug resistance profile/determinants differ depending on the lineage (ST) of S. argenteus.

Staphylococcus argenteus, a novel staphylococcal species independent of S. aureus, causes a wide spectrum of infectious diseases. As detection of this species from humans and animals has been increasingly reported worldwide, its growing virulence and drug resistance via external genetic determinants has become concerning. In this study, the prevalence and genetic characteristics of virulence factors and drug resistance determinants were investigated for 82 S. argenteus clinical isolates in Hokkaido, Japan, for a one-year period starting in August 2019. These S. argenteus isolates corresponded to 0.66% of the total number of S. aureus isolates collected in the same period. The most prevalent genotype was sequence type (ST) 2250 and staphylocoagulase (coa) genotype XId (45.1%, n = 37), followed by ST1223-coa XV (30.5%, n = 25) and ST2198-coa XIV (24.4%, n = 20). Panton-Valentine leukocidin genes (lukS-PV-lukF-PV) were identified in a single ST2250 isolate. Only ST1223 isolates had the enterotoxin gene cluster (egc-2), seb, and selw (detection rate; 100%, 60%, and 84%, respectively), while sec, sey, sel26-sel27, tst-1 were only detected in ST2250 isolates (detection rate; 10.8%, 100%, 67.6%, and 10.8%, respectively). ST2198 isolates harbored selx at a significantly higher rate (60%) than isolates of other STs. Although most of S. argenteus isolates were susceptible to antimicrobials examined, ST2198 showed higher resistance rates to penicillin, macrolides, and aminoglycosides than other STs, and it harbored various resistance genes such as blaZ, erm(C), msr(A), lnuA, and aac(6′)-Ie-aph(2″)-Ia. Only one ST2250 isolate possessed SCCmec-IVc, showing resistance to oxacillin. blaZ was the most prevalent determinant of resistance in the three STs and belonged to two plasmid groups and a chromosomal group, suggesting its diverse origin. lnu(A) in ST2198 isolates was assigned to a major cluster with various staphylococcal species. The present study indicates that the prevalence of virulence factors and drug resistance profile/determinants differ depending on the lineage (ST) of S. argenteus.

Objectives Methicillin-resistant Staphylococcus aureus (MRSA) is one of the leading causes of bloodstream infections (BSIs). We aimed to study molecular epidemiological characteristics of MRSA isolates from BSIs in northern Japan to elucidate the recent trend of their clonal diversity. Methods MRSA isolates (n = 277) were collected from blood samples of patients who attended healthcare facilities in Hokkaido, the northern main island of Japan, for a two-year period from August 2017. Genotypes, virulence factors/drug-resistance determinants, and structure of SCCmec complex were analysed by PCR and sequencing analysis. Results SCCmec-IIa (n = 171, 61.7%) with coagulase genotype (coa-) II, ST5/ST764/ST2389 was the most common genetic trait, followed by SCCmec-IVa (n = 78, 28.2%), and IVl (n = 10, 3.6%). Among the MRSA-IVa, 14 isolates (5.1% of all the isolates) had genetic features identical to USA300 clone (ST8/coa-IIIa/spa-t008 having ΦSa2USA and ACME-I), while PVL/ACME-negative MRSA-IVa isolates (n = 64) were classified into coa-IIa/IIIa/VIIa/VIIb, with coa-VIIa/spa-t1784/ST1 being dominant. Other minor clones included ST8-SCCmec-I, and ST30/ST45/ST81/ST121/ST1232-SCCmec-V, among which the ST1232 isolate harboured PVL genes. Spermidine N-acetyltransferase gene (speG), which is typically present in ACME-I of USA300 clone, was also identified in two isolates, ACME-II’-positive ST764-MRSA-IIa and ACME-negative ST1-MRSA-IVa, showing resistance to spermine. speG of these isolates was located in additional SCCs adjacent to SCCmec. Conclusions Our present study revealed clonal diversity of MRSA from BSIs in Japan, with increased prevalence of ST8-USA300. Distinct types of speG-carrying SCCs associated with SCCmec-II or IV were identified.

Objectives Clostridium perfringens is a common anaerobic pathogen causing enteritis/enterocolitis and wound infections in humans. We analyzed clonal diversity and toxin gene prevalence in C. perfringens clinical isolates from humans in northern Japan. Methods Prevalence of nine toxin genes was analyzed for 585 C. perfringens isolates from patients collected for 20-month period between May 2019 and December 2020 by molecular methods. Sequence type (ST) based on multilocus sequence typing (Xiao's scheme) and alpha-toxin (PLC) sequence type were determined for a total of 124 isolates selected in the present study along with those in our previous study (2017–2018). Results Toxinotypes A (68.2%) was the most frequent, followed by F (31.6%), and G (0.2%), while additional toxin genes encoding binary enterotoxin (BEC/CPILE) and beta2 toxin were identified in one and six isolates, respectively. Among the 124 isolates with various toxin gene profiles, 62 STs including 53 novel types were identified, revealing the presence of six clonal complexes (CCs) consisting of 27 STs. Most of enterotoxin gene (cpe)-positive isolates belonged to CC36, CC41, and CC117. Based on 22 key amino acids in alpha toxin sequence, four PLC types (I-IV) including 21 subtypes were classified, and their relation to individual STs/CCs was clarified. Two isolates harboring bec/cpile belonged to different STs (ST95, ST131) and PLC types (If, IVb), indicating distribution of this toxin gene to distinct lineages. Conclusions The present study revealed the diversity in C. perfringens clones of human origin with various toxin gene profiles represented by ST/CC and PLC type.

Although bio-oils produced by pyrolysis and hydro thermal synthesis demonstrate potential toward building a sustainable society, large amounts of char generated as a byproduct and their thermal instability owing to high oxygen content hinder their applications. Hence, a novel approach for the production of high-grade bio-oil was proposed herein. In this approach, zerovalent Fe was used as an agent for generating hydrogen in situ in the hydrothermal liquefaction of oil palm empty fruit bunch (EFB), a lignocellulosic biomass source, affording bio-oil containing water-soluble (WS) and water-insoluble (WI) fractions in high yields. Hydrogen generated by the reaction between Fe and H2O efficiently converted unstable intermediates obtained from the degradation of EFB into stable compounds, resulting in reduced char formation. Hydroxyketones were detected as components characteristic of the WS fraction in the H2O/EFB/Fe system, which were stable under hydrothermal condition. WS fractions were treated with the HZSM-5 zeolite, affording light olefins (C-2-C-4), as well as benzene, toluene, and xylene. This conversion was more efficient with the WS fraction obtained in the presence of Fe. The liquefaction of EFB and the conversion of WS fractions into olefins via catalytic cracking were also achieved using recycled Fe.

Prevalence and genetic characteristics of methicillin-resistant Staphylococcus aureus (MRSA) and coagulase-negative staphylococci in oral cavity of healthy children were studied in Hokkaido, northern main island of Japan. From saliva of 526 children, a total of 248 staphylococcal isolates comprising S. aureus (n = 143), S. epidermidis (n = 84), S. warneri (n = 13), S. haemolyticus (n = 5), S. hominis (n = 2), and S. intermedius (n = 1) were recovered. Presence of mecA was confirmed in 6.3% of S. aureus, 50% of S. epidermidis, and 7.7% in S. warneri. SCCmec was mostly classified into type IV, and ACME (arginine catabolic mobile element)-arcA was detected in S. epidermidis (23.8%) and S. intermedius. Nine MRSA isolates belonged to staphylocoagulase gene (coa) type Ia, IIa, IIIa, VIIb/sequence type 1 (ST1), ST5, ST8, ST89, ST120, and were negative for PVL (Panton-Valentine leukocidin) genes. These isolates included two clones of emerging community-acquired MRSA (CA-MRSA) that had been described recently in Japan: ST5/SCCmec IVc, which resembles the "Pediatric clone," and ST8/SCCmec IVl belonging to coa-IIIa/agr-I with sasL gene, designated "CA-MRSA/J" clone. Various enterotoxin genes were found in all the MRSA and some methicillin-susceptible S. aureus (MSSA) isolates examined, while tst-1 was detected in four MRSA isolates. Notably, a variant of elastin-binding protein gene (ebpS-v) was identified in ST120 MRSA and ST45 MSSA isolates, and exfoliative toxin D gene (etd) was detected in an MSSA isolate. The present study revealed the presence of MRSA, including the novel CA-MRSA clones, and high prevalence of methicillin-resistant S. epidermidis in oral cavity of healthy children in Japan.

Hydrogenation and hydrogenolysis reactions, which are used in the chemical industry for the synthesis of organic compounds, are very expensive operations because of the need for facilities that can liquefy, transport, and store the hydrogen produced through steam reforming of natural gas. We have therefore developed a novel approach for hydrogenation that does not require the use of high-cost facilities. Using this, zerovalent iron (Fe) and water (H 2 O) are introduced as an in situ hydrogen donor system for the transformation of methyl laurate into lauryl alcohol over a Ru-based catalyst. This combination of a Ru–Sn–Mo/C catalyst with a Fe/H 2 O system showed significantly higher transformation rates for the conversion of methyl laurate into lauryl alcohol than a conventional reaction system that uses pressurized hydrogen. The reason for this is that the new system produces lauric acid as an intermediate during the reaction, which is more efficiently hydrogenized into lauryl alcohol over the Ru–Sn–Mo/C catalyst. The Fe/H 2 O system played two important roles: a hydrogen source for the hydrogenation reaction and a catalyst for the generation of lauric acid by methyl laurate hydrolysis.

Purpose This crossover study was carried out to evaluate the influence of two different types of fluoridated dentifrices containing sodium fluoride (NaF) and sodium monofluorophosphate (MFP) on salivary fluoride levels after toothbrushing. Methods Eight consenting individuals (average age 36.4 years) brushed their teeth in the same manner using a paste type of dentifrices (0.5 g) containing 1500 ppm F. Following toothbrushing for 2 min, they spat out once and rinsed their mouths with 15 mL of distilled water for 5 s. Resting saliva samples were collected for 3 min each at different time intervals; 5 min, 10 min, 15 min, 30 min, 60 min, 120 min, and 180 min following the mouth rinsing. Fluoride concentrations in saliva samples were determined using a fluoride ion specific electrode. Then we compared those of dentifrices, in regard to salivary fluoride concentrations in each individual. The area under the curve (AUC) of fluoride was calculated using a graph on which the horizontal base line represents sampling time and the vertical scale represents fluoride concentration. Results Statistical differences ( p p Conclusion The results indicated that the NaF type of dentifrice retains more fluoride in saliva than that of MFP.

ツバキ葉抽出物500および2000mg/kgをラットに単回または28日間反復経口投与し，安全性を検討した．単回投与試験において，いずれの群にも死亡例はなく，一般状態，体重および解剖検査で異常がないことが確認された．また，28日間反復経口投与試験においても，ツバキ葉抽出物投与群に死亡例はなく，一般状態，体重，摂餌量，尿検査，血液学的検査，血液生化学的検査，器官重量，解剖検査および病理組織学的検査で異常は認められなかった．以上のことから，本実験条件ではツバキ葉抽出物500mg/kgと2000mg/kgのいずれにおいても経口摂取による毒性は検出されなかった．さらに，ツバキ葉抽出物の遺伝子突然変異誘発性の有無を，Salmonella typhimurium TA100，TA98，TA1535およびTA1537ならびにEscherichia coli WP2uvrAを用いたプレインキュベーション法による変異原性試験により検討した．ツバキ葉抽出物処理群における復帰変異コロニー数は，いずれの菌株ともS9mix添加の有無にかかわらず，陰性対照の2倍未満であり，本実験条件ではツバキ葉抽出物から突然変異原性は検出されなかった．

Twenty male volunteers, average age 24 years, participated in this study. Specimens were obtained by enamel biopsy using 5 microliters of 0.5 M HClO4 for 30 s. Using a regression curve, comparisons of fluoride concentrations were made at different depths. The fluoride concentrations (mean +/- SE) at a depth of 5 microns were highest in the distobuccal (1698 +/- 136), high in the mesiobuccal (1343 +/- 122), low in the distolingual (1119 +/- 107), and lowest in the mesiolingual sites (819 +/- 78). Of the interior enamels (or = 10 microns in depth), the distobuccal site (1330 +/- 88 parts/10(6) F at 10 microns) had a higher-concentration than all other sites. The fluoride profiles were steepest to shallowest in the order: distobuccal, mesiobuccal, distolingual and mesiolingual. There were no correlations between the enamel fluoride concentrations and the fluoride concentration in parotid saliva. It was concluded that in vivo fluoride profiles of maxillary first molars reflect the wear of the tooth surface with age and the condition of dental plaque deposition, and, to some extent, the site-specific distribution of saliva between buccal and lingual surfaces.

Previous studies showed that strontium (Sr) as well as fluoride (F) can enhance enamel remineralisation. The aim of this study was to evaluate the effects of Sr in combination with F on enamel remineralisation in vitro. Sixty enamel specimens obtained from caries free human premolars were demineralised to produce caries-like lesions. Half of each lesion was covered with nail varnish as an untreated control. The specimens were then randomly divided into F and Sr + F treatment groups. The F group was exposed to remineralising solutions (1.5 mM CaCl2, 0.9 mM KH2PO4) containing 1 ppm, 0.1 ppm or 0.05 ppm F. The Sr + F treatment group was exposed to the same solutions including 10 ppm Sr. After 2 weeks, lesion depth, mineral loss and percentage enamel remineralisation were determined using transversal microradiography. There was a significant decrease in mineral loss in all groups (p < 0.001). Lesion depth was significantly reduced for all groups (p < 0.05) with the exception of group F. Remineralisation was significantly affected by F concentration (p = 0.000). The participation of Sr resulted in a significant enhancement of remineralisation (p < 0.001) with a synergistic effect of the Sr + F combination (p < 0.01). It was concluded that while the remineralising process was affected by the concentration of F, there was also an interaction between F and Sr when they were used in conjunction.