Effects of ozone nano-bubble water on periodontopathic bacteria and oral cells - in vitro studies.

The aims of the present study were to evaluate the bactericidal activity of a new antiseptic agent, ozone nano-bubble water (NBW3), against periodontopathogenic bacteria and to assess the cytotoxicity of NBW3 against human oral cells. The bactericidal activities of NBW3 against representative periodontopathogenic bacteria, Porphyromonas gingivalis (P. gingivalis) and Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) were evaluated using in vitro time-kill assays. The cytotoxicity of NBW3 was evaluated using three-dimensional human buccal and gingival tissue models. The numbers of colony forming units (CFUs)/mL of P. gingivalis and A. actinomycetemcomitans exposed to NBW3 dropped to below the lower limit of detection (<10 CFUs mL-1) after only 0.5 min of exposure. There were only minor decreases in the viability of oral tissue cells after 24 h of exposure to NBW3. These results suggest that NBW3 possesses potent bactericidal activity against representative periodontopathogenic bacteria and is not cytotoxic to cells of human oral tissues. The use of NBW3 as an adjunct to periodontal therapy would be promising.

A case-control study evaluating relative risk factors for decompression sickness: a research report.

BACKGROUND: Factors contributing to the pathogenesis of decompression sickness (DCS) in divers have been described in many studies. However, relative importance of these factors has not been reported. METHODS: In this case-control study, we compared the diving profiles of divers experiencing DCS with those of a control group. The DCS group comprised 35 recreational scuba divers who were diagnosed by physicians as having DCS. The control group consisted of 324 apparently healthy recreational divers. All divers conducted their dives from 2009 to 2011. The questionnaire consisted of 33 items about an individual's diving profile, physical condition and activities before, during and just after the dive. To simplify dive parameters, the dive site was limited to Izu Osezaki. Odds ratios and multiple logistic regression were used for the analysis. RESULTS: Odds ratios revealed several items as dive and health factors associated with DCS. The major items were as follows: shortness of breath after heavy exercise during the dive (OR = 12.12), dehydration (OR = 10.63), and maximum dive depth > 30 msw (OR = 7.18). Results of logistic regression were similar to those by odds ratio analysis. CONCLUSION: We assessed the relative weights of the surveyed dive and health factors associated with DCS. Because results of several factors conflict with previous studies, future studies are needed.

Clinical and microbiological effects of ozone nano-bubble water irrigation as an adjunct to mechanical subgingival debridement in periodontitis patients in a randomized controlled trial.

AIM: Ozone nano-bubble water (NBW3) seems to be suitable as an adjunct to periodontal treatment owing to its potent antimicrobial effects, high level of safety, and long storage stability. The aim of the present study was to evaluate the clinical and microbiological effects of NBW3 irrigation as an adjunct to subgingival debridement for periodontal treatment. METHODS: Twenty-two subjects were randomly assigned to one of the two treatment groups: full-mouth mechanical debridement with tap water (WATER) or full-mouth mechanical debridement with NBW3 (NBW3). Clinical examination was performed at baseline and 4 and 8 weeks after treatment. Microbiological examination was carried out just before and after treatment and at 1 and 8 weeks posttreatment. RESULTS: There were significant improvements in all clinical parameters after 4 weeks in both groups. The reduction in the probing pocket depth and the clinical attachment gain after 4 and 8 weeks in the NBW3 group were significantly greater than those in the WATER group. Moreover, only the NBW3 group showed statistically significant reductions in the mean total number of bacteria in subgingival plaque over the study period. CONCLUSIONS: The present study suggests that subgingival irrigation with NBW3 may be a valuable adjunct to periodontal treatment. CLINICAL RELEVANCE: This study verified the potential of new antimicrobial agent, MNW3, as an adjunct to periodontal treatment.

The pathway of auxin biosynthesis in plants.

The plant hormone auxin, which is predominantly represented by indole-3-acetic acid (IAA), is involved in the regulation of plant growth and development. Although IAA was the first plant hormone identified, the biosynthetic pathway at the genetic level has remained unclear. Two major pathways for IAA biosynthesis have been proposed: the tryptophan (Trp)-independent and Trp-dependent pathways. In Trp-dependent IAA biosynthesis, four pathways have been postulated in plants: (i) the indole-3-acetamide (IAM) pathway; (ii) the indole-3-pyruvic acid (IPA) pathway; (iii) the tryptamine (TAM) pathway; and (iv) the indole-3-acetaldoxime (IAOX) pathway. Although different plant species may have unique strategies and modifications to optimize their metabolic pathways, plants would be expected to share evolutionarily conserved core mechanisms for auxin biosynthesis because IAA is a fundamental substance in the plant life cycle. In this review, the genes now known to be involved in auxin biosynthesis are summarized and the major IAA biosynthetic pathway distributed widely in the plant kingdom is discussed on the basis of biochemical and molecular biological findings and bioinformatics studies. Based on evolutionarily conserved core mechanisms, it is thought that the pathway via IAM or IPA is the major route(s) to IAA in plants.

The AMI1 gene family: indole-3-acetamide hydrolase functions in auxin biosynthesis in plants.

Novel genes that function in the conversion of indole-3-acetamide (IAM) into indole-3-acetic acid (IAA), which were previously thought to exist only in the bacterial genome, have been isolated from plants. The finding of the AtAMI1 gene in Arabidopsis thaliana and the NtAMI1 gene in Nicotiana tabacum, which encode indole-3-acetamide hydrolase, indicates the existence of a new pathway for auxin biosynthesis in plants. This review summarizes the characteristics of these genes involved in auxin biosynthesis and discusses the possibility of the AMI1 gene family being widely distributed in the plant kingdom. Its evolutionary relationship to bacterial indole-3-acetamide hydrolase, based on phylogenetic analyses, is also discussed.

The effects of hyperbaric oxygen on tooth movement into the regenerated area after distraction osteogenesis.

OBJECTIVE: To analyze the effect of hyperbaric oxygen on newly formed bone in distracted areas surrounding the root of a moving tooth by histological and radiological analysis. It was hypothesized that the application of hyperbaric oxygen to a tooth moving into the distracted area would accelerate ossification and vascularization of newly formed bone in the distracted space. DESIGN: Ten dogs were used. After creating a 10-mm-long bone defect, a bony segment was prepared and translocated into the defect area at a rate of 1 mm/d for 10 days. Following the distraction period, tooth movement was started and the dogs were divided into two groups. The HBO group received hyperbaric oxygen; whereas, the control group did not. At 150 days after tooth movement, the distracted area around the moving tooth was evaluated radiologically and histologically. Differences between groups were confirmed by a Mann-Whitney U test. RESULTS: Trabecular bone density and cortical and subcortical bone areas measured by peripheral quantitative computed tomography in the HBO group were significantly higher than those in the control group. Histological observations revealed regenerated bone and blood vessels formation in the tension site of the moving tooth in the HBO group. The regenerated bone structure measured by bone histomorphometry was larger and more active in bone formation in the HBO group than in the control group. CONCLUSIONS: Applying hyperbaric oxygen to tooth movement into a distracted area appears to accelerate ossification and vascularization of regenerated bone in the that area.

Enhancement of glucose toxicity by hyperbaric oxygen exposure in diabetic rats.

The side effects of hyperbaric oxygen (HBO) treatment, such as oxidative stress and oxygen toxicity, have long been of interest. However, there are no comprehensive studies evaluating such toxic effects in diabetes mellitus (DM). The purpose of this study was to determine the effects of HBO on glucose homeostasis and histological changes in pancreatic beta-cells of experimentally induced diabetic rats. A total of 24 male Wistar rats were randomly divided into 4 groups: 1) Control group, no diabetic induction without HBO treatment; 2) HBO group, exposed to 100% oxygen at 2.8 ATA (atmosphere absolute) for 2 h once daily, for 7 days; 3) DM group, diabetes induced by streptozotocin (STZ) injection; and 4) DM + HBO group, received both STZ injection and HBO exposure. HBO treatment, with clinically recommended pressures and duration of therapy, was started on day 5 after STZ injection, when the blood glucose levels were significantly increased. After the last HBO treatment, the pancreatic tissues were immunostained to measure the areas of insulin immunoreactive beta-cells in the islets of Langerhans. The blood glucose increased significantly following exposure to HBO, with the highest levels achieved in rats, which had been treated with both HBO and diabetic induction. The area populated with insulin immunoreactive beta-cells decreased significantly following diabetic induction and/or HBO exposure, with the smallest area in DM + HBO group. Thus, HBO exposure enhanced the cytotoxic effect of STZ in the beta-cells of the pancreas. HBO should be cautiously employed in diabetic patients.

[Acute decompression illness following hyperbaric exposure: clinical features of central nervous system involvement].

Decompression illness (DCI) is a general term encompassing all pathological changes secondary to reduction of environmental pressure. This condition has two forms: decompression sickness (DCS) and arterial gas embolism (AGE) secondary to pulmonary barotrauma. Moreover, DCS is categorized as minor, such as limb and/or joint pains or skin rash (Type I), and serious, as in cardiopulmonary and/or central nervous system involvements (Type II). Cerebral and spinal injuries have been symptomatically classified into AGE and DCS. Brain scans of patients with AGE or DCS showed multiple cerebral infarctions in the terminal and/or border zones of the cerebral arteries. Brain involvements of patients in both AGE and DCS show no differences in neurological or neuro-radiological findings. From the neurological and radiological standpoint, it is therefore impossible to distinguish these two conditions. Despite established treatments for neurological DCI (both AGE and DCS), it is unclear whether US Navy treatment Table 6 is preferable to standard hyperbaric oxygen therapy such as 2.4 atmospheres pressure for 90 minutes. Japanese laws and regulations have peculiarities that permit air diving to 90 meters depth, but with explicit prohibition of the use of oxygen for decompression, albeit a limited use of mixed gas is permissible. Moreover, currently the health screening for hyperbaric workers does not include detailed examination of the cardiopulmonary or the central nervous system.

[Preservation of quality of life and its relation to lifestyle in centenarians in Japan: a visitation interview survey].

PURPOSE: To investigate quality of life (QOL) and associated factors in centenarians, we conducted a visitation interview survey for all male and one half of the female centenarians living in Japan, and studied QOL, lifestyle and health status. METHODS: All the male centenarians and a randomly-sampled half of the female centenarians living in Japan were included in the study based on the National Census. Finally, 556 male and 1341 female centenarians participated in the present visitation interview survey and were studied using a structured questionnaire. QOL was assessed from 3 aspects; activities of daily life (ADL), cognitive function (CF) and mental health (MH). RESULTS: Individuals having superior ADL accounted for 30.9% of the male and 13.6% of the female centenarians, those with superior CF were 58.1% and 35.6%, respectively, and those with superior MH were 39.2% and 26.8%, providing evidence that superior QOL was more dominant in males than in females. There were no effects of residential areas (north, east, middle, west, southwest and south of Japan) on the prevalence of ADL, CF and MH. Multiple logistic regression analyses conducted independently, for ADL, CF and MH as dependent valuables revealed that habitual exercise, less decline in visual ability and preservation of masticating ability were associated with better results for these 3 aspects of male centenarians' QOL, and that habitual exercise, less decline in visual ability, spontaneous awakening in the morning, good appetite and having cohabitating family members were associated with superior QOL in the females, suggesting that more factors are operating in the latter. CONCLUSIONS: Several lifestyle factors were found to be associated with preservation of QOL in centenarians in Japan. These results should facilitate establishing a public health and welfare policy for the most senior citizens, a group that has been increasing in Japan.

The NtAMI1 gene functions in cell division of tobacco BY-2 cells in the presence of indole-3-acetamide.

Tobacco (Nicotiana tabacum) Bright Yellow-2 (BY-2) cells can be grown in medium containing indole-3-acetamide (IAM). Based on this finding, the NtAMI1 gene, whose product is functionally equivalent to the AtAMI1 gene of Arabidopsis thaliana and the aux2 gene of Agrobacterium rhizogenes, was isolated from BY-2 cells. Overexpression of the NtAMI1 gene allowed BY-2 cells to proliferate at lower concentrations of IAM, whereas suppression of the NtAMI1 gene by RNA interference (RNAi) caused severe growth inhibition in the medium containing IAM. These results suggest that IAM is incorporated into plant cells and converted to the auxin, indole-3-acetic acid, by NtAMI1.

Function of the aux and rol genes of the Ri plasmid in plant cell division in vitro.

Auxin-autonomous growth in vitro may be related to the integration and expression of the aux and rol genes from the root-inducing (Ri) plasmid in plant cells infected by agropine-type Agrobacterium rhizogenes. To elucidate the functions of the aux and rol genes in plant cell division, plant cell lines transformed with the aux1 and aux2 genes or with the rolABCD genes were established using tobacco (Nicotiana tabacum) Bright Yellow-2 (BY-2) cells. The introduction of the aux1 and aux2 genes enabled the auxin-autonomous growth of BY-2 cells, but the introduction of the rolABCD genes did not affect the auxin requirement of the BY-2 cells. The results clearly show that the aux genes are necessary for auxinautotrophic cell division, and that the rolABCD genes are irrelevant in auxin autotrophy.

The aux1 gene of the Ri plasmid is sufficient to confer auxin autotrophy in tobacco BY-2 cells.

Tobacco (Nicotiana tabacum) Bright Yellow-2 (BY-2) cells are rapidly proliferating meristematic cells that require auxin for culture in vitro. We have established several transgenic BY-2 cell lines that carry the T-DNA of Agrobacterium rhizogenes 15834, which harbors an agropine-type root-inducing (Ri) plasmid. Two of these lines, BYHR-3 and BYHR-7, were used to test the role of auxin in the proliferation of plant cells. The lines grew rapidly in Linsmaier-Skoog (LS) medium lacking auxin and other phytohormones. The TR-DNA, containing the aux1 (tryptophan monooxygenase) and aux2 (indoleacetamide hydrolase) genes, was present in the genomes of both transgenic lines, whereas the TL-DNA, containing the rolA, B, C and D genes, was present in the genome of BYHR-7 but not BYHR-3. Since the introduction of the rolABCD genes alone did not affect the auxin requirement of BY-2 cells, the aux1 and aux2 genes, but not the rolABCD genes, appear to be relevant to the auxin autotrophy of these transgenic lines. Furthermore, the overexpression of aux1 allowed BY-2 cells to grow rapidly in the absence of auxin, suggesting the existence in plant cells of an unidentified gene whose product is functionally equivalent or similar to that of aux2 of the Ri plasmid.

Hyperbaric oxygen induces basic fibroblast growth factor and hepatocyte growth factor expression, and enhances blood perfusion and muscle regeneration in mouse ischemic hind limbs.

BACKGROUND: It is not clear how hyperbaric oxygen therapy (HBO) affects ischemia-induced pathophysiological responses such as angiogenesis and skeletal muscle regeneration. In the present study the effects of HBO on the functional and morphological recovery of ischemic hind limbs, blood perfusion and the local production of angiogenic growth factors were studied in a mouse model. METHODS AND RESULTS: Mice were placed in pure oxygen under 3 atm for 1 h/day for 14 days after the removal of a segment of the left femoral artery. HBO-treated mice showed better functional recovery and greater blood flow in the ischemic hind limb than untreated mice. Histological examination revealed unatrophied muscle fibers with islands of small regenerating muscle cells only in HBO-treated mice. Regeneration of muscle was confirmed by the increase in myf5 mRNA. The amount of mRNA for vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF) and basic fibroblast growth factor (bFGF) was slightly increased in the ischemic hind limbs. HBO eliminated the increase in VEGF mRNA. In contrast, the amount of mRNA for bFGF and HGF was further increased by HBO treatment. HBO transiently increased early growth response protein 1 (Egr-1) in the ischemic hind limbs. CONCLUSIONS: HBO accelerates the recovery of ischemic hind limbs by increasing the production of bFGF and HGF and by promoting muscle regeneration in mice.

Early progression from dimethyl sulfoxide-induced G(0)/G(1) arrest in L(1210) cells.

Recently, dimethyl sulfoxide (DMSO) has been used as a convenient cryoprotectant for stem cells in stem cell transplantation using allogenic peripheral blood or umbilical cord blood. As the stem cells have a multipotency, clarification of the extent of cell proliferation after transplantation is difficult. In the present study, DMSO gradually induced G(0)/G(1) arrest in mouse leukemia L(1210) cells with good cell viability. After removal of DMSO, the cells proliferated appropriately, resulting in expression of the DNA-synthesizing enzymes thymidylate synthase and thymidine kinase within 6h, and the cells entering into S phase within 12h. The sequence was followed by the marked activation of both enzymes within 24h and the increase of bromodeoxyuridine (BrdU) immunoreactive (S phase) cells with rapid cell proliferation within 36 h. In conclusion, mouse leukemia L(1210) cells, which were treated with 1.5% DMSO for 96 h, tolerated the treatment and reversed the cell cycle arrest within 36 h.

Growth-promoting effect of carbon material upon bacterial cells propagating through a distance.

Carbon material such as graphite and activated charcoal, but not diamond, causes the promotion of growth of certain bacteria under ordinarily non-permissive stress conditions over a distance of several centimeters. Bacillus carboniphilus under the stress of a high KCl concentration and high temperature responded to this remote effect of carbon material with enhanced growth, and thermophile bacterium Bacillus stearothermophilus responded similarly yet moderately under the stress of low temperature. The remote effect of carbon was caused by its activation with external energy, probably of electromagnetic nature, as this effect was markedly decreased by sheltering the experimental system with an iron or aluminum barrier. Carbon material probably transforms the external oscillatory pulses or radiation into a signal exerting, far-reaching, growth-promoting effect upon cells. The most plausible candidate of signals emitted from carbon was considered to be (ultra)sonic.

Production of sound waves by bacterial cells and the response of bacterial cells to sound.

Bacterial cells enhance the proliferation of neighboring cells under stress conditions by emitting a physical signal. Continuous single sine sound waves produced by a speaker at frequencies of 6-10, 18-22, and 28-38 kHz promoted colony formation by Bacillus carboniphilus under non-permissive stress conditions of high KCl concentration and high temperature. Furthermore, sound waves emitted from cells of Bacillus subtilis at frequencies between 8 and 43 kHz with broad peaks at approximately 8.5, 19, 29, and 37 kHz were detected using a sensitive microphone system. The similarity between the frequency of the sound produced by B. subtilis and the frequencies that induced a response in B. carboniphilus and the previously observed growth-promoting effect of B. subtilis cells upon B. carboniphilus through iron barriers, suggest that the detected sound waves function as a growth-regulatory signal between cells.