Complete genome sequence of Opitutales bacterium strain ASA1, isolated from soil.

We present the complete genome of Opitutales bacterium ASA1, isolated from soil. The genome is 5,821,695 bp with 4,638 protein-coding sequences. The genome data suggest that this strain belongs to the class Opitutae of the phylum Verrucomicrobiota, and its genome has six unique biosynthetic gene clusters associated with secondary metabolites.

Transcriptional Regulation of Methanol Dehydrogenases in the Methanotrophic Bacterium Methylococcus capsulatus Bath by Soluble and Insoluble Lanthanides.

The effects of soluble and insoluble lanthanides on gene expression in Methylococcus capsulatus Bath were investigated. Genes for lanthanide-containing methanol dehydrogenases (XoxF-MDHs) and their calcium-containing counterparts (MxaFI-MDHs) were up- and down-regulated, respectively, by supplementation with soluble lanthanide chlorides, indicating that M. capsulatus has the "lanthanide switch" observed in other methanotrophs. Insoluble lanthanide oxides also induced the lanthanide switch and were dissolved by the spent medium of M. capsulatus, suggesting the presence of lanthanide-chelating compounds. A transcriptome ana-lysis indicated that a gene cluster for the synthesis of an enterobactin-like metal chelator contributed to the dissolution of insoluble lanthanides.

Construction of an autocatalytic reaction cycle in neutral medium for synthesis of life-sustaining sugars.

Autocatalytic mechanisms in carbon metabolism, such as the Calvin cycle, are responsible for the biological assimilation of CO2 to form organic compounds with complex structures, including sugars. Compounds that form C-C bonds with CO2 are regenerated in these autocatalytic reaction cycles, and the products are concurrently released. The formose reaction in basic aqueous solution has attracted attention as a nonbiological reaction involving an autocatalytic reaction cycle that non-enzymatically synthesizes sugars from the C1 compound formaldehyde. However, formaldehyde and sugars, which are the substrate and products of the formose reaction, respectively, are consumed in Cannizzaro reactions, particularly under basic aqueous conditions, which makes the formose reaction a fragile sugar-production system. Here, we constructed an autocatalytic reaction cycle for sugar synthesis under neutral conditions. We focused on the weak Brønsted basicity of oxometalate anions such as tungstates and molybdates as catalysts, thereby enabling the aldol reaction, retro-aldol reaction, and aldose-ketose transformation, which collectively constitute the autocatalytic reaction cycle. These bases acted on sugar molecules of substrates together with sodium ions of a Lewis acid to promote deprotonation under neutral conditions, which is the initiation step of the reactions forming an autocatalytic cycle, whereas the Cannizzaro reaction was inhibited. The autocatalytic reaction cycle established using this abiotic approach is a robust sugar production system. Furthermore, we found that the synthesized sugars work as energy storage substances that sustain microbial growth despite their absence in nature.

A critical role of the periplasm in copper homeostasis in Gram-negative bacteria.

Copper is essential for life, but is toxic in excess. Copper homeostasis is achieved in the cytoplasm and the periplasm as a unique feature of Gram-negative bacteria. Especially, it has become clear the role of the periplasm and periplasmic proteins regarding whole-cell copper homeostasis. Here, we addressed the role of the periplasm and periplasmic proteins in copper homeostasis using a Systems Biology approach integrating experiments with models. Our analysis shows that most of the copper-bound molecules localize in the periplasm but not cytoplasm, suggesting that Escherichia coli utilizes the periplasm to sense the copper concentration in the medium and sequester copper ions. In particular, a periplasmic multi-copper oxidase CueO and copper-responsive transcriptional factor CusS contribute both to protection against Cu(I) toxicity and to incorporating copper into the periplasmic components/proteins. We propose that Gram-negative bacteria have evolved mechanisms to sense and store copper in the periplasm to expand their living niches.

Self-cloning of the Catalase Gene in Environmental Isolates Improves Their Colony-forming Abilities on Agar Media.

Hydrogen peroxide (H2O2) inhibits microbial growth at a specific concentration. However, we previously isolated two environmental bacterial strains that exhibited sensitivity to a lower H2O2 concentration in agar plates. Putative catalase genes, which degrade H2O2, were detected in their genomes. We herein elucidated the characteristics of these putative genes and their products using a self-cloning technique. The products of the cloned genes were identified as functional catalases. The up-regulation of their expression increased the colony-forming ability of host cells under H2O2 pressure. The present results demonstrated high sensitivity to H2O2 even in microbes possessing functional catalase genes.

Genome Sequences of Comamonadaceae Bacteria OS-1 and OS-4, Two Highly H2O2-Sensitive Strains Isolated from Pond Water.

The Comamonadaceae bacterial strains OS-1 and OS-4 were isolated from pond water and were found to be highly sensitive to hydrogen peroxide in the agar plates. Here, we report the nearly complete and complete genome sequences, respectively, of these two strains.

Implant Placement Using the Bone Ring Technique With and Without Membrane Placement: A Preclinical 12-Month Study in Dogs.

The bone ring technique is applied for vertical augmentation, in which the dental implant and autogenous block bone graft are simultaneously inserted. We investigated the healing of bone around implants placed simultaneously using the bone ring technique with and without membrane placement after a 12-month healing period. Vertical bone defects were created on both sides of the mandible of Beagle dogs. Implants were inserted into the defects through bone rings and fixed with membrane screws as healing caps. The augmented sites on one side of the mandible were covered with a collagen membrane. Samples were harvested 12 months after implantation and assessed histologically and by microcomputed tomography analysis. All implants remained throughout the healing period; however, except for 1 implant, they showed lost caps and/or exposure to the oral cavity. The implants had contact with newly formed bone despite frequent bone resorption. The surrounding bone appeared mature. The medians of bone volume and percentages of total bone area and bone-to-implant contact within the bone ring were slightly higher in the group with membrane placement than in that without membrane placement. Nevertheless, none of the evaluated parameters were significantly affected by the membrane placement. In the present model, soft tissue complication was frequent, and the membrane application did not reveal the effect at 12 months after implant placement using the bone ring technique. Sustained osseointegration and maturation of surrounding bone were observed in both groups after a 12-month healing period.

Double-Crown Prosthesis Retention Using Polyetherketoneketone (PEKK): An In Vitro Study.

PURPOSE: To assess the retentive force of telescopic crowns using polyetherketoneketone (PEKK) high-performance polymer in relation to conventional materials over a long period of time in an in vitro setting. MATERIALS AND METHODS: Thirty-six sets of primary and secondary crowns were fabricated as per the double crown-retained prostheses approach. Six samples were included in each of the five test groups (1: zirconia/PEKK [Zr/PEKK]; 2: titanium/PEKK [Ti/PEKK]; 3: cobalt-chrome/PEKK [CoCr/PEKK]; 4: PEKK/PEKK; and 5: gold/PEKK [Au/PEKK]) and the single control group (gold/galvano-gold [Au/GA]). The insertion-removal test was performed for 20,000 cycles, and the surface condition was observed. Retentive forces were analyzed using two-way ANOVA (α<0.05). RESULTS: The retention forces in groups Zr/PEKK and Ti/PEKK significantly decreased over time (group 1: p = 0.035 and group 2: p = 0.001), whereas retentive force increased significantly in groups PEKK/PEKK, Au/PEKK, and control (group 4: p = 0.001, group 5: p = 0.008, and control: p = 0.042). Similar wear was observed on the primary crown in groups PEKK/PEKK, gold/PEKK, and control. CONCLUSIONS: Groups PEKK/PEKK and Au/PEKK showed a transition of retentive force similar to the control group. Groups PEKK/PEKK and Au/PEKK had similar wear on the surface compared to control. Therefore, PEKK has a promising clinical potential.

Critical Effect of H2O2 in the Agar Plate on the Growth of Laboratory and Environmental Strains.

We previously showed that autoclaving in preparing agar media is one of the sources of hydrogen peroxide (H2O2) in the medium. This medium-embedded H2O2 was shown to lower the total colony count of environmental microorganisms. However, the critical concentrations of H2O2 detrimental to colony formation on the agar plate remain largely undetermined. Herein, we elucidated the specific effect of H2O2 on microbial colony formation on solid agar medium by external supplementation of varying amounts of H2O2. While common laboratory strains (often called domesticated microbes) formed colonies in the presence of high H2O2 concentrations (48.8 µM or higher), microbes from a freshwater sample demonstrated greatly decreased colony counts in the presence of 8.3 µM H2O2. This implies that environmental microbes are susceptible to much lower concentrations of H2O2 than laboratory strains. Among the emergent colonies on agar plates supplemented with different H2O2 concentrations, the relative abundance of betaproteobacterial colonies was found to be lower on plates containing higher amounts of H2O2. Further, the growth of the representative betaproteobacterial isolates was completely inhibited in the presence of 7.2 µM H2O2. Therefore, our study clearly demonstrates that low micromolar levels of H2O2 in agar plates critically affect growth of environmental microbes, and large portions of those are far more susceptible to the same than laboratory strains. IMPORTANCE It is well-known that most of environmental microorganisms do not form colonies on agar medium despite that agar medium is the commonly used solidified medium. We previously demonstrated the negative effects of H2O2 generation during agar medium preparation on colony formation. In the present study, we investigated the independent effect of H2O2 on microbial growth by adding different concentrations of H2O2 to agar medium. Our results demonstrate for the first time that even low micromolar levels of H2O2 in agar plates, that are far lower than previously recognized as significant, adversely affect colony number obtained from freshwater inoculum.

Draft Genome Sequence of the Alkaliphilic, Lithoautotrophic Homoacetogen Fuchsiella alkaliacetigena Z-7100T.

Fuchsiella alkaliacetigena is a spore-forming, alkaliphilic hydrogentrophic homoacetogen that was isolated from the soda lake Lake Tanatar III in Russia. The genome of the type strain Z-7100 (= DSM 24880) is 2.9 Mb, with a G+C content of 36.2%.

Draft Genome Sequence of the Extremely Haloalkaliphilic and Homoacetic Bacterium Natroniella acetigena Z-7937T.

Natroniella acetigena Z-7937T (= DSM 9952T) is a heterotrophic homoacetogenic natronophile. The draft genome sequence is 2.6 Mb in 116 contigs, with a G+C content of 34.1%.

Peri-implant tissue augmentation by volume-stable collagen matrix transplantation: a study of dog mandibles.

The aim of this study was to investigate histologically the amount of peri-implant tissue augmentation after volume-stable porcine collagen matrix transplantation. Six male beagle dogs were used in the experiment. P2, P4, and M1 distal roots were extracted under general anesthesia. After 6 months, implants were placed in the same sites, and volume-stable porcine collagen matrix transplantation was performed. Impressions were taken at 1 and 2 weeks and at 1, 2, and 3 months after transplantation. The dogs were euthanized at 3 months, and their mandibles were removed and scanned using micro-computed tomography. Standard Triangulated Language data were also obtained. Using preoperative models as a reference, the data for all time points were compared, and changes in the thickness of the cross-section of the implant sites were measured. The model created at 3 months was then compared with the mandible data, and the thickness of collected peri-implant soft tissue was measured under optical microscopy. Increased thickness was found at some of the sites on the buccal side. Regarding the peri-implant soft tissue, the thickness of the measured sites on the buccal side was significantly increased at 3 months in the experimental group. Histological observations of the internal structures of the tissue in the experimental group revealed irregular collagen fibers and a remnant collagen matrix. Endogenous tissue was observed within the collagen matrix, indicating good fusion with the surrounding autologous tissue. These results suggest that volume-stable porcine collagen matrix transplantation promotes peri-implant tissue augmentation on the buccal side.

Vertical eruption of anterior maxillary teeth adjacent to single-implant-supported crowns: An assessment after a 3-year follow-up period.

STATEMENT OF PROBLEM: Vertical eruption of teeth adjacent to an implant has been reported clinically and might affect the esthetic outcome over time. The prevalence of the problem is unclear. PURPOSE: The purpose of this prospective clinical trial was to evaluate the vertical eruption of anterior maxillary teeth adjacent to single-implant crowns after a 3-year follow-up period. MATERIAL AND METHODS: Thirty single dental implants were inserted in maxillary anterior sites including the first premolar. The mean age of the participants at implant-supported crown insertion was 48.4 years (range 23 to 79 years). Each implant was restored with a 1-piece screw-retained ceramic single crown. The vertical changes of 60 adjacent anterior maxillary teeth were evaluated from periapical radiographs and casts at baseline, 6, 12, and 36 months. RESULTS: A global ANOVA test showed statistically significant differences for the outcomes of the distance-implant platform and cement-enamel junction (DPC) (P<.001), crown length distal (P=.021) and mesial (P=.035), implant crown length (P=.022), and incisal edge to edge (P<.001). CONCLUSIONS: Continuous vertical tooth eruption next to a single dental implant was observed in adult participants.

Complete Genome Sequence of Alphaproteobacteria Bacterium Strain SO-S41, Isolated from Forest Soil.

The complete genome of hydrogen peroxide-sensitive alphaproteobacterial strain SO-S41 was sequenced. The complete genome contains a single chromosome, is 4,443,179 bp in length, contains a total of 4,632 genes, and has a G+C content of 66.2%.

Retention forces between primary and secondary CAD/CAM manufactured telescopic crowns: an in vitro comparison of common material combinations.

OBJECTIVES: To analyze the retention forces between primary and secondary telescopic crowns milled from various materials and to compare them with the retention forces between cast telescopic crowns made of precious metal alloys. MATERIALS AND METHODS: Primary and secondary crowns (N = 60; n = 10 per group) were fabricated using various material combinations (1: zirconia [ZIR]/polyether ether ketone [PEEK]; 2: titanium grade IV [TI]/PEEK; 3: PEEK/PEEK; 4: non-precious alloy [NPA]/PEEK; 5:NPA/NPA), while precious alloy (PA) was used for the control group (6: PA/PA). The retention forces at 10, 1000, 5000, and 10,000 connection and disconnection cycles and the relative weights were analyzed, applying nonparametric repeated measures ANOVA and post hoc Mann-Whitney and Wilcoxon signed-rank tests (α < 0.05). RESULTS: Globally, significant differences in the retention forces among the materials (p < 0.0001), time points (p < 0.0001), and wear resistance for the various materials (p < 0.0001) were observed. No significant changes in retention forces compared to baseline were observed in groups 2, 4, 5, and 6. A significantly higher weight loss for both primary and secondary crowns was observed in groups 4 and 6. CONCLUSIONS: The material combination in telescopic attachments influences retention forces and wear. Interactions between materials and time were evident, indicating that the change in retention forces differs among the materials. The combinations of milled TI/PEEK and NPA/NPA qualify for further preclinical testing in a more clinically realistic setup, determining a material-specific double-crown design. CLINICAL RELEVANCE: The design of precious alloy telescopic crowns cannot be directly transferred to other milled material combinations due to different retention behaviors.

CBCT imaging and histopathological characteristics of osteoradionecrosis and medication-related osteonecrosis of the jaw.

PURPOSE: The purpose of this study was to evaluate the cone-beam computed tomographic (CBCT) imaging and histopathological characteristics of osteoradionecrosis (ORN) and medication-related osteonecrosis of the jaw (MRONJ). MATERIALS AND METHODS: Ten surgical specimens from segmental mandibulectomy (3 ORN and 7 MRONJ) were analyzed using CBCT. The CBCT parameters were as follows: high-resolution mode (tube voltage, 90.0 kV; tube current, 4.00 mA; rotation time, 16.8 s; field of view, 56 mm×56 mm; thickness, 0.099 mm). Histopathological characteristics were evaluated using histological slides of the surgical specimens. The Pearson chi-square test was used to compare ORN and MRONJ in terms of CBCT findings (internal texture, sequestrum, periosteal reaction and cortical perforation) and histopathological characteristics (necrotic bone, inflammatory cells, reactive bone formation, bacteria, Actinomyces, and osteoclasts). A P value less than 0.05 was considered to indicate statistical significance. RESULTS: MRONJ showed periosteal reaction on CBCT more frequently than ORN (7 of 7 [100%] vs. 0 of 3 [0%], P<0.05). Regarding histopathological characteristics, MRONJ showed osteoclasts more frequently than ORN (6 of 7 [85.7%] vs. 0 of 3 [0%], P<0.05). CONCLUSION: This study evaluated the CBCT imaging and histopathological characteristics of ORN and MRONJ, and the findings suggest that CBCT could be useful for the evaluation of ORN and MRONJ.

Reductive Transformation of Fe(III) (oxyhydr)Oxides by Mesophilic Homoacetogens in the Genus Sporomusa.

Microbial reduction of iron contributes to the dissolution and transformation of iron-containing minerals in nature. Diverse groups of homoacetogenic bacteria (homoacetogens) have been reported to reduce insoluble Fe(III) oxides, such as hydrous ferric oxide (HFO), an Fe(III) mineral commonly found in soils and sediments. Several members of genus Sporomusa reportedly oxidize Fe(0), indicating the presence of an extracellular electron-uptake mechanism. However, the ability of the genus to reduce insoluble Fe(III) oxides is limited, and the underlying reduction mechanism remains to be elucidated. In this study, the HFO reduction ability of three Sporomusa spp. (Sporomusa sp. strain GT1, Sporomusa sphaeroides, and Sporomusa ovata) and a homoacetogen of a different genus (Acetobacterium woodii) were assayed under organotrophic (ethanol) and lithotrophic (H2 + CO2) conditions without a chelator or reducing reagent. All tested homoacetogens showed acetogenic growth and concomitant reduction of HFO under both organotrophic and lithotrophic conditions. Analysis of the growth stoichiometry showed that Fe(III) reduction does not support direct energy conservation, thereby indicating that Fe(III) reduction is a side reaction of acetogenesis to dissipate the excess reducing power. HFO was reduced to a soluble Fe(II) form by microbial activity. In addition, we observed that strain GT1, S. sphaeroides, and S. ovata reduced crystalline Fe(III) oxides, and HFO was reductively transformed into magnetite (Fe3O4) under phosphate-limiting conditions. Separation of HFO by a dialysis membrane still permitted Fe(II) production, although the reduction rate was decreased, suggesting that Fe(III) reduction is at least partially mediated by soluble redox compound(s) secreted from the cells. Finally, culture experiments and comparative genomic analysis suggested that electron transfer by flavins and multiheme c-type cytochrome were not directly correlated with Fe(III) reduction activity. This study reveals the capability of Sporomusa spp. in the reductive transformation of iron mineral and indicates the potential involvement of these organisms in iron and other mineral cycles in nature.

Single versus splinted short implants at sinus augmented sites: A systematic review and meta-analysis.

OBJECTIVE: This review primarily evaluated the success, survival and failure rates of implants shorter than 10 mm restored with single-unit or splinted fixed dental prostheses in maxillary sinus augmented sites. MATERIAL AND METHODS: Two reviewers independently performed the systematic search of electronic databases, including MEDLINE, EMBASE and CENTRAL, up to September 2019 with no language restriction. A supplemental hand search consisted of screening 13 journals. The inclusion criteria were: primary studies reporting implant, prosthetic and patient-reported outcome measures (PROMs) of extra-short and short implants placed in conjunction with sinus floor elevation in partially dentate patients, restored with single- and splinted-crowns for direct comparison, with a minimal 1-year follow-up. Weighted arithmetic mean (WAM) of the implant survival was performed according to the type of prosthesis. This was confirmed by using Review Manager software to perform meta-analysis. RESULTS: Two observational studies reporting on 106 tapered, press-fit, sintered porous-surfaced implants with a length ranging from 5 mm to 9 mm were included in this systematic review. Of these, 20 and 86 implants were restored with single and splinted prostheses, respectively. The risk ratio (RR) was 1.16 (95% CI: .31-4.30, p = .58, I² = 0%) for individually restored implants failure when compared to splinted implants, indicating that short dental implants restored with single crowns could have a 16% higher possibility of failure if compared to implants with splinted crowns. The heterogeneity value was not statistically significative (p = .58). No statistical difference in the implant survival rate of the two types of analysed prostheses was observed after WAM (p= .923). The level of evidence for the included studies ranged from low (4) to fair (2B). CONCLUSION: Similar clinical outcomes up to a 9-year follow-up were observed in single and splinted porous-surfaced implants shorter than 10 mm located in sites with sinus lift. However, the conclusion shall be interpreted with caution due to the level of evidence and limited number of included studies included in this systematic review.

Variability in tongue pressure among elderly and young healthy cohorts: A systematic review and meta-analysis.

OBJECTIVES: Tongue pressure (TP) is used for the diagnosis of oral hypofunction; however, the impact of several variables on TP is unclear. Therefore, the current systematic review and meta-analysis aimed to analyse the variability in tongue pressure among healthy individuals aged ≥60 years vs <60 years. Secondary outcomes were the influence of gender and the type of measuring device (Iowa Oral Performance Instrument (IOPI) vs JMS tongue pressure measurement device (JMS)). METHODS: PubMed and the Japanese database Ichushi-Web were searched systematically by two independent reviewers for studies reporting TP values in healthy populations. Clinical studies published between 1959 and June 2020 with more than 10 participants, written in English, German or Japanese, were included. A random-effects meta-regression for aggregate-level data was applied (α < 0.05). RESULTS: Sixty-eight studies reported TP for a total of 13 773 subjects aged <60 years (n = 3265) and ≥60 years (n = 10 508). TP was significantly higher in subjects <60 years (estimated weighted mean (EWM) ± standard error = 51.9 ± 1.28 kPa; 95% CI = 49.4-54.4) relative to those ≥60 (EWM = 34.7 ± 0.94 kPa; 95% CI = 32.8-36.5) (P < .001), men (EWM = 45.9 ± 2.09 kPa; 95% CI = 41.8-50.0) relative to women (EWM = 39.3 ± 1.68 kPa; 95% CI = 36.0-42.6) (P = .015) and when assessed with the IOPI (EWM = 51.9 ± 1.32 kPa; 95% CI = 49.3-54.5) compared to the JMS (EWM = 33.5 ± 0.63 kPa; 95% CI = 32.2-34.7) (P < .001). In terms of gender, there was no significant difference in TP among subjects ≥60 years (P = .282). However, in subjects younger than 60, a significant difference was observed (P = .004). CONCLUSIONS: Healthy populations aged <60 years showed significantly higher TP than those aged ≥60 years. TP values ascertained by the IOPI are significantly higher than those obtained with the JMS.

An iron corrosion-assisted H2-supplying system: a culture method for methanogens and acetogens under low H2 pressures.

H2 is an important fermentation intermediate in anaerobic environments. Although H2 occurs at very low partial pressures in the environments, the culture and isolation of H2-utilizing microorganisms is usually carried out under very high H2 pressures, which might have hampered the discovery and understanding of microorganisms adapting to low H2 environments. Here we constructed a culture system designated the "iron corrosion-assisted H2-supplying (iCH) system" by connecting the gas phases of two vials (one for the iron corrosion reaction and the other for culturing microorganisms) to achieve cultures of microorganisms under low H2 pressures. We conducted enrichment cultures for methanogens and acetogens using rice paddy field soil as the microbial source. In the enrichment culture of methanogens under canonical high H2 pressures, only Methanobacterium spp. were enriched. By contrast, Methanocella spp. and Methanoculleus spp., methanogens adapting to low H2 pressures, were specifically enriched in the iCH cultures. We also observed selective enrichment of acetogen species by the iCH system (Acetobacterium spp. and Sporomusa spp.), whereas Clostridium spp. predominated in the high H2 cultures. These results demonstrate that the iCH system facilitates culture of anaerobic microorganisms under low H2 pressures, which will enable the selective culture of microorganisms adapting to low H2 environments.