Comprehensive microbiological findings in peri-implantitis and periodontitis.

AIM: The microbial differences between peri-implantitis and periodontitis in the same subjects were examined using 16S rRNA gene clone library analysis and real-time polymerase chain reaction. MATERIALS AND METHODS: Subgingival plaque samples were taken from the deepest pockets of peri-implantitis and periodontitis sites in six subjects. The prevalence of bacteria was analysed using a 16S rRNA gene clone library and real-time polymerase chain reaction. RESULTS: A total of 333 different taxa were identified from 799 sequenced clones; 231 (69%) were uncultivated phylotypes, of which 75 were novel. The numbers of bacterial taxa identified at the sites of peri-implantitis and periodontitis were 192 and 148 respectively. The microbial composition of peri-implantitis was more diverse when compared with that of periodontitis. Fusobacterium spp. and Streptococcus spp. were predominant in both peri-implantitis and periodontitis, while bacteria such as Parvimonas micra were only detected in peri-implantitis. The prevalence of periodontopathic bacteria was not high, while quantitative evaluation revealed that, in most cases, prevalence was higher at peri-implantitis sites than at periodontitis sites. CONCLUSIONS: The biofilm in peri-implantitis showed a more complex microbial composition when compared with periodontitis. Common periodontopathic bacteria showed low prevalence, and several bacteria were identified as candidate pathogens in peri-implantitis.

Lysyl oxidase like-2 reinforces unsatisfactory ossification induced by bone morphogenetic protein-2: relating nanomechanical properties and molecular changes.

Bone morphogenetic protein-2 (BMP2) is among the most popular anabolic agents and substantially increase bone volume related to enhanced osteoblast differentiation. Here we demonstrate a remarkable deterioration in the nanomechanical properties of mineralized tissue induced from osteoblasts solely by the function of BMP2. Mineralized tissue of primary osteoblasts cultured with BMP2 shows molecular features of both bone and cartilage, but depletion of lysyl oxidase family members leads to poor nanomechanical properties of the mineralized tissue. Lysyl oxidase like-2 supplementation reinforces the inferior mineralized tissue induced from osteoblasts by BMP2 through intermolecular cross-linking of type II or type X collagen-rich extracellular matrix. This may also mimic a consolidation of bone fracture gaps, despite the fact that the distribution of the bone properties in such microenvironments has been poorly elucidated. These findings confirm the importance of testing newly induced bone down to the microscale and nanoscale in bone tissue engineering. FROM THE CLINICAL EDITOR: Bone morphogenetic protein-2 is known to substantially increase bone volume related to enhanced osteoblast differentiation; however, this team of investigators report a remarkable deterioration in the nanomechanical properties of mineralized tissue induced from osteoblasts solely by the function of BMP2.

Complete Genome Sequence of Tea Plant Necrotic Ring Blotch Virus Detected from a Tea Plant in Japan.

We report the complete genome sequence of a Japanese isolate of Tea plant necrotic ring blotch virus (TPNRBV-J). The predicted TPNRBV-J genes have the same organization as those of a Chinese isolate, and the 5' termini of the segments have conserved nucleotide sequences.

Nationwide survey of salmonella prevalence in environmental dust from layer farms in Japan.

A nationwide survey was conducted to determine Salmonella prevalence in airborne dust from layer farms. Of the 4,090 layer farms in Japan, 203 were surveyed and 48 (23.6%) of these were positive for Salmonella. Salmonella isolation rates were higher in the eastern (24.3%), central (25.6%), western (23.9%), and southern (27.5%) prefectures than they were in the northern (13.3%) prefecture. We recovered 380 Salmonella isolates and identified 34 different Salmonella serovars. Salmonella Infantis was the most prevalent serovar (42 [11.1%] of 380), followed by Salmonella Agona (39 [10.3%] of 380), Salmonella Mbandaka (37 [9.7%] of 380), Salmonella Cerro (32 [8.4%] of 380), Salmonella Thompson (29 [7.6%] of 380), and Salmonella Braenderup (27 [7.1%] of 380). Of the 380 isolates, 273 (71.8%) were resistant to more than one antibiotic. Salmonella Infantis (41 [97.6%] of 42), Salmonella Agona (38 [97.4%] of 39), and Salmonella Mbandaka (34 [91.9%] of 37) showed the highest resistance rates. We found 18 different resistance patterns and the most common (179 [47.1%] of 273) was resistant to dihydrostreptomycin. One of the 13 Salmonella Hadar isolates was resistant to eight antibiotics. To investigate characteristics of Salmonella Agona, Salmonella Infantis, and Salmonella Mbandaka isolates across different prefectures, we performed pulsed-field gel electrophoresis by using XbaI and BlnI. The Salmonella Agona and Salmonella Mbandaka dendrograms were grouped into seven clusters, with 80 and 70% similarity, respectively. Because the Salmonella Infantis dendrogram showed low similarity, there is a possibility of genetic diffusion of this serovar across Japan. This report is the first to describe Salmonella contamination in airborne dust from layer farms in Japan. Our findings should be useful for future Salmonella infection monitoring and control.

Intra-strain biological and epidemiological characterization of plum pox virus.

Plum pox virus (PPV) is one of the most important plant viruses causing serious economic losses. Thus far, strain typing based on the definition of 10 monophyletic strains with partially differentiable biological properties has been the sole approach used for epidemiological characterization of PPV. However, elucidating the genetic determinants underlying intra-strain biological variation among populations or isolates remains a relevant but unexamined aspect of the epidemiology of the virus. In this study, based on complete nucleotide sequence information of 210 Japanese and 47 non-Japanese isolates of the PPV-Dideron (D) strain, we identified five positively selected sites in the PPV-D genome. Among them, molecular studies showed that amino acid substitutions at position 2,635 in viral replicase correlate with viral titre and competitiveness at the systemic level, suggesting that amino acid position 2,635 is involved in aphid transmission efficiency and symptom severity. Estimation of ancestral genome sequences indicated that substitutions at amino acid position 2,635 were reversible and peculiar to one of two genetically distinct PPV-D populations in Japan. The reversible amino acid evolution probably contributes to the dissemination of the virus population. This study provides the first genomic insight into the evolutionary epidemiology of PPV based on intra-strain biological variation ascribed to positive selection.

Complete Genome Sequence of Iris Severe Mosaic Virus Isolated in Japan.

The complete genome sequence of an iris severe mosaic virus isolate (ISMV) from Iris tectorum in Japan was determined for the first time. According to sequence identity analyses, our specimen is closely related to isolates reported from China.

Distinct interacting core taxa in co-occurrence networks enable discrimination of polymicrobial oral diseases with similar symptoms.

Polymicrobial diseases, which can be life threatening, are caused by the presence and interactions of multiple microbes. Peri-implantitis and periodontitis are representative polymicrobial diseases that show similar clinical symptoms. To establish a means of differentiating between them, we compared microbial species and functional genes in situ by performing metatranscriptomic analyses of peri-implantitis and periodontitis samples obtained from the same subjects (n = 12 each). Although the two diseases differed in terms of 16S rRNA-based taxonomic profiles, they showed similarities with respect to functional genes and taxonomic and virulence factor mRNA profiles. The latter-defined as microbial virulence types-differed from those of healthy periodontal sites. We also showed that networks based on co-occurrence relationships of taxonomic mRNA abundance (co-occurrence networks) were dissimilar between the two diseases. Remarkably, these networks consisted mainly of taxa with a high relative mRNA-to-rRNA ratio, with some showing significant co-occurrence defined as interacting core taxa, highlighting differences between the two groups. Thus, peri-implantitis and periodontitis have shared as well as distinct microbiological characteristics. Our findings provide insight into microbial interactions in polymicrobial diseases with unknown etiologies.

Increased visceral fat and serum levels of triglyceride are associated with insulin resistance in Japanese metabolically obese, normal weight subjects with normal glucose tolerance.

OBJECTIVE: The purpose of this study was to investigate the association between visceral adiposity or triglyceride (TG) metabolism and insulin resistance in metabolically obese, normal weight (MONW) Japanese individuals with normal glucose tolerance. RESEARCH DESIGN AND METHODS: We evaluated body fat areas, lipid profiles, and the glucose infusion rate (GIR) during a euglycemic-hyperinsulinemic clamp study in 20 MONW subjects (BMI <25 kg/m(2)and visceral fat areas 100 cm(2)) with normal glucose tolerance. Body fat areas were measured by computed tomography scans. Control data were obtained from 20 normal subjects (BMI <25 kg/m(2) and visceral fat areas <100 cm(2)). RESULTS: MONW subjects showed a significant increase in fasting serum levels of TG (P < 0.01) and a decrease in GIR (P < 0.01) compared with normal subjects. There were significant correlations between visceral fat areas (r = -0.563, P < 0.01) or serum levels of TG (r = -0.474, P < 0.05) and GIR in MONW subjects. Multiple regression analyses showed that visceral fat areas (F = 7.702, P < 0.02) and serum levels of TG (F = 7.114, P < 0.05) were significantly associated with GIR in all (MONW and normal) subjects. CONCLUSIONS: Increased visceral fat and serum levels of TG are associated with insulin resistance in Japanese MONW subjects with normal glucose tolerance. Excess visceral fat and elevated TG levels may play important roles in the development of insulin resistance in Japanese MONW subjects with normal glucose tolerance.

A review of improved fixation methods for dental implants. Part II: biomechanical integrity at bone-implant interface.

PURPOSE: The purpose of this article is to review the mechanical requirements of the tissue-implant interface and analyze related theories. STUDY SELECTION: The osseointegration capacity of titanium implants has been investigated over the past 50 years. We considered the ultimate goal of osseointegration to which form a desirable interfacial layer and a bone matrix with adequate biomechanical properties. RESULTS: Occasionally, the interface comprises porous titanium and bone ingrowth that enables a functionally graded Young's modulus, thereby allowing reduction of stress shielding. However, the optimal biomechanical connection at the interface has not yet been fully clarified. There have been publications supporting several universal mechanical testing technologies in terms of bone-titanium bonding ability, although the separation of newly formed bone quality is unlikely. CONCLUSIONS: The understanding of complex mechanical bone behavior and size-dependent properties ranging from a nano- to a macroscopic level are essential in the biomechanical optimization of implants. The requirements of regenerated tissue at the interface include high strength, fracture toughness related to ductility, and time-dependent energy dissipation and/or elastic-plastic stress distribution. Moreover, a strong relationship between strain signals and peri-implant tissue turnover could be expected, so that ideal implant biomechanics may enable longevity via adaptive bone remodeling.

Bone micro-fragility caused by the mimetic aging processes in α-klotho deficient mice: in situ nanoindentation assessment of dilatational bands.

The nanoscale structure-function relationship is a key determinant of bone toughness or micro-fragility. The loss of bone toughness during the aging process has been accepted based on empirical evidence, but this concept has not yet been fully supported by evidence at the material level. Here, we demonstrate a reduction in bone toughening mechanism in mimetic aged cortical bone obtained from α-klotho deficient (α-klotho(-/-)) mice and assessed by in situ dynamic mechanical analysis. The strain-rate nanoindentation tests showed enhanced stiffening of the wild-type calvarial bone and a large dimensional recovery during rapid loading following the constant displacement test. Such strain-dependent stiffening was likely associated with nanoscale dilatational bands and subsequent strain-energy transfer to the superior wild-type cross-linked collagen matrix network. The absence of dilatational bands formed by hydroxyapatite crystals and non-collagenous proteins in the α-klotho(-/-) bone samples likely diminished the intrinsic bone toughening mechanisms almost independent of viscoelastic behaviors. Such nanoscale structural alternations that occur during aging processes lead to crack propagation and result in overall bone fractures under large external stresses. In addition, dynamic mechanical analysis using instrumented nanoindentation was useful for the evaluation of bone mechanical properties in this pathological model of a genetic knockout mouse.

Oxidative stress is associated with adiposity and insulin resistance in men.

To investigate the direct relationship of oxidative stress with obesity and insulin resistance in men, we measured the plasma levels of 8-epi-prostaglandin F2alpha (PGF2alpha) in 14 obese and 17 nonobese men and evaluated their relationship with body mass index; body fat weight; visceral, sc, and total fat areas, measured by computed tomography; and glucose infusion rate during a euglycemic hyperinsulinemic clamp study. Obese men had significantly higher plasma concentrations of 8-epi-PGF2alpha than nonobese men (P < 0.05). The plasma levels of 8-epi-PGF2alpha were significantly correlated with body mass index (r = 0.408; P < 0.05), body fat weight (r = 0.467; P < 0.05), visceral (r = 0.387; P < 0.05) and total fat area (r = 0.359; P < 0.05) in all (obese and nonobese) men. There was also a significant correlation between the plasma levels of 8-epi-PGF2alpha and glucose infusion rate in obese men (r = -0.552; P < 0.05) and all men (r = -0.668; P < 0.01). In all subjects, the plasma levels of 8-epi-PGF2alpha were significantly correlated with fasting serum levels of insulin (r = 0.487; P < 0.01). In brief, these findings showed that the circulating levels of 8-epi-PGF2alpha are related to adiposity and insulin resistance in men. Although correlation does not prove causation, the results of this study suggest that obesity is an important factor for enhanced oxidative stress and that this oxidative stress triggers the development of insulin resistance in men.

Nitric oxide stimulates glucose transport through insulin-independent GLUT4 translocation in 3T3-L1 adipocytes.

OBJECTIVE: It is well known that nitric oxide synthase (NOS) is expressed and that it modulates glucose transport in skeletal muscles. Recent studies have shown that adipose tIssues also express inducible and endothelial nitric oxide synthase (eNOS). In the present study, we investigated whether nitric oxide (NO) induces glucose uptake in adipocytes, and the signaling pathway involved in the NO-stimulated glucose uptake in 3T3-L1 adipocytes. METHODS: First, we determined the expression of eNOS in 3T3-L1 adipocytes, and then these cells were treated with the NO donor sodium nitroprusside (SNP) and/or insulin, and glucose uptake and phosphorylation of insulin receptor substrate (IRS)-1 and Akt were evaluated. Moreover, we examined the effects of a NO scavenger, a guanylate cyclase inhibitor or dexamethasone on SNP-stimulated glucose uptake and GLUT4 translocation. RESULTS: SNP at a concentration of 50 mmol/l increased 2-deoxyglucose uptake (1.8-fold) without phosphorylation of IRS-1 and Akt. Treatment with the NO scavenger or guanylate cyclase inhibitor decreased SNP-stimulated glucose uptake to the basal level. Dexamethasone reduced both insulin- and SNP-stimulated glucose uptake with impairment of GLUT4 translocation. CONCLUSION: NO is capable of stimulating glucose transport through GLUT4 translocation in 3T3-L1 adipocytes, via a mechanism different from the insulin signaling pathway.

Pineal cystic germinoma with syncytiotrophoblastic giant cells mimicking MR imaging findings of a pineal cyst.

We report a case of precocious puberty in a 4-year-old boy. Contrast-enhanced T1-weighted MR imaging suggested a pineal cyst with enhancement of the slightly thickened wall and focal wall irregularity. Three-dimensional constructive interference in a steady-state imaging revealed a focal lobulation and a nodule-like area in the lesion. The lesion mimicking a pineal cyst proved to be a germinoma with syncytiotrophoblastic giant cells, on the basis of biopsy and tumor marker results.

Intraindividual variation in core microbiota in peri-implantitis and periodontitis.

The oral microbiota change dramatically with each part of the oral cavity, even within the same mouth. Nevertheless, the microbiota associated with peri-implantitis and periodontitis have been considered the same. To improve our knowledge of the different communities of complex oral microbiota, we compared the microbial features between peri-implantitis and periodontitis in 20 patients with both diseases. Although the clinical symptoms of peri-implantitis were similar to those of periodontitis, the core microbiota of the diseases differed. Correlation analysis revealed the specific microbial co-occurrence patterns and found some of the species were associated with the clinical parameters in a disease-specific manner. The proportion of Prevotella nigrescens was significantly higher in peri-implantitis than in periodontitis, while the proportions of Peptostreptococcaceae sp. and Desulfomicrobium orale were significantly higher in periodontitis than in peri-implantitis. The severity of the peri-implantitis was also species-associated, including with an uncultured Treponema sp. that correlated to 4 clinical parameters. These results indicate that peri-implantitis and periodontitis are both polymicrobial infections with different causative pathogens. Our study provides a framework for the ecologically different bacterial communities between peri-implantitis and periodontitis, and it will be useful for further studies to understand the complex microbiota and pathogenic mechanisms of oral polymicrobial diseases.

Strain-rate stiffening of cortical bone: observations and implications from nanoindentation experiments.

While bone mineralization is considered to be responsible for its stiffness, bone durability partially associated with the time-dependent viscoelasticity of matrix proteins is still poorly elucidated. Here we demonstrate a novel mechanism of highly mineralized bone durability almost independent of inherent viscoelastic behaviour along with a protocol for measuring the mechanical properties of mineralized tissues. Strain-rate nanoindentation tests showed substantial stiffening of the highly mineralized calvarial bone, whereas large creep or stress relaxation was observed during constant load or displacement tests, respectively. Based on the lower viscoelasticity of the highly mineralized structure, such large time-dependent response appears to be associated with nanoscale dimensional recovery, rather than viscoelastic behaviour, implying the inverse namely strain-rate dependent dilatant behaviour. This dilatant expansion increased the indenter penetration resistance into the surface, enhancing instantaneous stiffness. The associated stiffening and higher effective elastic modulus were highly strain-rate dependent and more readily observed in more highly mineralized tissues such as the calvarial bone. Such strain-rate stiffening and consequent dimensional recovery may be vital responses of bone tissues against excessive deformation to maintain tissue integrity.

Comparison of the prevalence of shiga toxin-producing Escherichia coli strains O157 and O26 between beef and dairy cattle in Japan.

With the aim of comparing the prevalence of Shiga toxin-producing Escherichia coli (STEC) O157 and O26 between beef and dairy cattle, we collected rectal content samples from 250 beef cattle on 25 beef farms and 250 dairy cows on 25 dairy farms from July through September 2011. STEC O157 was isolated from 16 beef cattle on 7 beef farms, while no STEC O157 was isolated from any dairy farms. This result suggests that the prevalence of STEC O157 is higher in beef cattle than in dairy cattle. STEC O26 was isolated from 1 animal each from beef and dairy cattle herds, and therefore, it was not possible to compare statistically the prevalence of STEC O26 in beef and dairy cattle.

Prevalence and characterization of foodborne pathogens in dairy cattle in the eastern part of Japan.

To investigate the prevalence and characterization of foodborne pathogens [Campylobacter spp., Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes and Salmonella spp.] in dairy cows, rectal content grab samples were collected from 250 dairy cows reared on 25 dairy farms in eastern Japan from December 2010 through February 2011. Campylobacter jejuni was isolated from 106 (42%) cows on 23 (92%) farms, STEC O157 from three cows on one farm, L. monocytogenes from three cows on another three farms and Salmonella enterica subsp. enterica serovar Typhimurium from eight cows on another farm. STEC O26 was not isolated from any of the dairy farms investigated. The results suggest that C. jejuni is widespread in dairy farms in eastern Japan.