Evolutionary Dynamics of Host Organs for Microbial Symbiosis in Tortoise Leaf Beetles (Coleoptera: Chrysomelidae: Cassidinae).

Diverse insects host specific microbial symbionts that play important roles for their growth, survival, and reproduction. They often develop specialized symbiotic organs for harboring the microbial partners. While such intimate associations tend to be stably maintained over evolutionary time, the microbial symbionts may have been lost or replaced occasionally. How symbiont acquisitions, replacements, and losses are linked to the development of the host's symbiotic organs is an important but poorly understood aspect of microbial symbioses. Cassidine leaf beetles are associated with a specific gammaproteobacterial lineage, Stammera, whose reduced genome is streamlined for producing pectin-degrading enzymes to assist the host's digestion of food plants. We investigated the symbiotic system of 24 Japanese cassidine species and found that (i) most species harbored Stammera within paired symbiotic organs located at the foregut-midgut junction, (ii) the host phylogeny was largely congruent with the symbiont phylogeny, indicating stable host-symbiont association over evolutionary time, (iii) meanwhile, the symbiont was not detected in three distinct host lineages, uncovering recurrent losses of the ancient microbial mutualist, (iv) the symbiotic organs were vestigial but present in the symbiont-free lineages, indicating evolutionary persistence of the symbiotic organs even in the absence of the symbiont, and (v) the number of the symbiotic organs was polymorphic among the cassidine species, either two or four, unveiling a dynamic evolution of the host organs for symbiosis. These findings are discussed as to what molecular mechanisms and evolutionary trajectories underpin the recurrent symbiont losses and the morphogenesis of the symbiotic organs in the herbivorous insect group. IMPORTANCE Insects represent the biodiversity of the terrestrial ecosystem, and their prosperity is attributable to their association with symbiotic microorganisms. By sequestering microbial functionality into their bodies, organs, tissues, or cells, diverse insects have successfully exploited otherwise inaccessible ecological niches and resources, including herbivory enabled by utilization of indigestible plant cell wall components. In leaf beetles of the subfamily Cassininae, an ancient symbiont lineage, Stammera, whose genome is extremely reduced and specialized for encoding pectin-degrading enzymes, is hosted in gut-associated symbiotic organs and contributes to the host's food plant digestion. Here, we demonstrate that multiple symbiont losses and recurrent structural switching of the symbiotic organs have occurred in the evolutionary course of cassidine leaf beetles, which sheds light on the evolutionary and developmental dynamics of the insect's symbiotic organs and provides a model system to investigate how microbial symbionts affect the host's development and morphogenesis and vice versa.

Bond durability and surface states of titanium, Ti-6Al-4V alloy, and zirconia for implant materials.

PURPOSE: Screw-retained implant crowns used as dental implants comprise a zirconia coping and titanium base bonded using resin cement. These devices are prone to debonding failures. This study investigated the bond characteristics of implant materials based on shear bond strength (SBS) and surface characteristics. METHODS: Chemically pure (CP) titanium grade-4 (Ti), Ti-6Al-4V alloy (Ti-6Al-4V), and tetragonal polycrystalline zirconia (zirconia) were evaluated as adherent materials. Plates of each material were polished, primed for the respective resin cements, and cemented using either methyl methacrylate-based resin cement (Super-Bond) or composite-based resin cement (Panavia). The cemented samples were subjected to 10,000 thermocycles alternating between 5 and 55 °C, and the SBS were obtained before and after thermocycling. The sample surfaces were characterized based on surface observations, roughness, and free energy (SFE). RESULTS: The SBSs of all materials bonded using Panavia were significantly compromised during thermocycling and reached zero. Although the SBSs of Ti and Ti-6Al-4V bonded using Super-Bond were not significantly affected by thermocycling, those of zirconia decreased significantly. The bond durability between zirconia and Super-Bond was improved via alumina air-abrasion, which caused no significant loss of SBS after thermocycling. Surface analyses of the air-abraded zirconia validated these results and confirmed that its surface roughness and SFE were significantly increased. CONCLUSION: The bond durability between resin cement and zirconia was lower than that between Ti and Ti-6Al-4V. The alumina air-abrasion pretreatment of zirconia improved the SFE and surface roughness, thereby enhancing bond durability.

The immature platelet fraction affects the efficacy of platelet rich plasma therapy for knee osteoarthritis.

INTRODUCTION: Platelet-rich plasma (PRP) therapy is used to treat pathological conditions such as degenerative inflammatory diseases including osteoarthritis (OA) by enhancing tissue repair and promoting anti-inflammatory effects. Although PRP therapy for patients with knee OA improved pain and functional scores, the association of clinical outcomes and quality of PRP including cell composition and concentration is unclear. METHODS: Therefore, this study analyzed blood cell counts, including the immature platelet fraction (IPF), in peripheral blood and PRP of 144 patients with knee OA who underwent PRP therapy. The mean leukocyte and platelet concentrations in whole blood and PRP were analyzed using an XN-1000 automated hematology analyzer. Visual analogue scale (VAS) scores and knee injury and osteoarthritis outcome scores (KOOS) before and 1 month after a single PRP injection were also determined. RESULTS: Higher platelet and lower leukocyte concentration rates were observed in PRP compared with whole blood. The platelet concentration in whole blood was negatively correlated with VAS improvement. The percentage of IPF (IPF%) in whole blood was positively correlated with VAS improvement and KOOS (pain) improvement, whereas the IPF% in PRP tended to correlate with VAS improvement. Furthermore, multivariate logistic regression demonstrated the high IPF% in whole blood was significantly associated with VAS improvement. The low percentage of neutrophil (neutrophil%) in PRP was significantly associated with the VAS improvement and KOOS (ADL) improvement. CONCLUSIONS: Therefore, PRP efficacy for OA might depend on the patient's biological status.

Artificial intelligence supported anemia control system (AISACS) to prevent anemia in maintenance hemodialysis patients.

Anemia, for which erythropoiesis-stimulating agents (ESAs) and iron supplements (ISs) are used as preventive measures, presents important difficulties for hemodialysis patients. Nevertheless, the number of physicians able to manage such medications appropriately is not keeping pace with the rapid increase of hemodialysis patients. Moreover, the high cost of ESAs imposes heavy burdens on medical insurance systems. An artificial-intelligence-supported anemia control system (AISACS) trained using administration direction data from experienced physicians has been developed by the authors. For the system, appropriate data selection and rectification techniques play important roles. Decision making related to ESAs poses a multi-class classification problem for which a two-step classification technique is introduced. Several validations have demonstrated that AISACS exhibits high performance with correct classification rates of 72%-87% and clinically appropriate classification rates of 92%-98%.

Effects of alumina airborne-particle abrasion on the surface properties of CAD/CAM composites and bond strength to resin cement.

The purpose of this study is to clarify physical and chemical changes in surfaces of CAD/CAM composites caused by alumina airborne-particle abrasion and its effect on adhesive bonding. Our study involved three dispersed filler (DF)-based and a polymer-infiltrated ceramic network (PICN)-based CAD/CAM composites. Changes in the surface morphologies of the composites were examined, and surface free energy (SFE) analysis was performed based on Owens-Wendt theory. The influence of the abrasion on the bond strengths of CAD/CAM composites to the resin cement was characterized by shear bond strength (SBS) test. The abrasion increased the roughness of the composites. The SFE analysis showed that the abrasion significantly increased the dispersive component but decreased the polar component of the SFE associated with the DF-based composites, while no change occurred for those of the PICN-based composite. The abrasion slightly improved the SBSs for the DF-based composites but not that of the PICN-based composite.

Adhesive bonding of alumina air-abraded Ag-Pd-Cu-Au alloy with 10-methacryloyloxydecyl dihydrogen phosphate.

The aim of this paper is to study changes in the Ag-Pd-Cu-Au alloy surfaces by alumina air-abrasion process and effect of those changes on the adhesive bonding characteristic. Surface roughness, surface composition and chemical state of the alumina air-abraded alloys were analyzed by a confocal laser scanning microscope, an energy dispersive X-ray spectroscopy and an X-ray photoelectron spectroscopy. The results showed that the alumina air-abrasion changed the alloy surface by mechanical roughening, alumina remain and copper oxidation. Effect of the changes in the alloy surface on the adhesive bonding characteristic was examined by using a methyl methacrylate/tri-n-butylborane derivative (MMA/TBB) resin cement with the 10-methacryloyloxydecyl dihydrogen phosphate (MDP) contained primer. The shear bond strength test results indicated that the surface oxidation by the abrasion is the main contributor that improved the adhesive bonding rather than other effects such as mechanical roughening or alumina remain.

Chemical alteration of Ag-Pd-Cu-Au alloy surface by alumina air-abrasion and its effect on bonding to resin cement.

The goal of this study was to investigate the chemical alteration of a dental alloy surface by alumina air-abrasion and its effect on bonding to resin cement. Alumina air-abrasion was carried out on an Ag-Pd-Cu-Au alloy. The surface morphology and chemical state of the abraded alloy were characterized. The effect of the air-abrasion on the shear bond strength between the alloy and a methyl methacrylate/tri-n-butyl borane (MMA/TBB) resin cement with some primers was evaluated. The surface characterization revealed that the alumina air-abrasion mechanically roughened and chemically altered the surface. The chemical alterations had two effects: (1) abraded alumina particles remained on the alloy surface and (2) copper ions were oxidized in the alloy surface. As the result, the shear bond strength test indicated that 10-methacryloyloxydecyl dihydrogen phosphate (MDP) contained primer worked with the abraded alloy surface, whereas it did not work with the non-abraded alloy surface.

Phytochemical characterization of Rosa multiflora Thunb. (Rosaceae) in Japan and South Korea, with a focus on the bioactive flavonol glycoside 'multiflorin A'.

Dried achene or anthocarpous accessory fruits of Rosa multiflora Thunb., Rosae fructus ("Eijitsu" in Japanese), have been used in clinical practice to improve constipation within traditional Japanese medicine. Recently, it has been claimed that the efficacy of this crude drug is decreasing, and multiflorin A, the purgative component, was not detected within the tested samples. In order to clarify the causes of this issue, we investigated Rosa section Synstylae (Rosaceae), including R. multiflora, growing in Japan and South Korea with a focus on the secondary metabolite, multiflorin A. We recognize that there are two chemotypes based on the presence (Type I) or absence (Type II) of multiflorin A. Type I contains quercitrin, multinoside A, multiflorin B, and multinoside A acetate as major index compounds. Type II contains hyperin, isoquercitrin, quercetin 3-O-glucuronide, and 3'-methoxy-isoquercitrin as the major index compounds. The chemotype of Rosa section Synstylae (Rosaceae) plants collected in Japan (excluding Tsushima Island) were all classified as Type I with exception of two species, R. luciae and R. sambucina. On the other hand, both Type I and Type II were detected within Rosae fructus obtained from R. multiflora collected in South Korea and Tsushima Island, Japan. The results indicate that Rosae fructus from R. multiflora (Type I) from Japan, excluding Tsushima Island, should be employed clinically, which we describe as purgative.

Determining auditory-evoked activities from multiple cells in layer 1 of the dorsal cortex of the inferior colliculus of mice by in vivo calcium imaging.

Layer 1 of the dorsal cortex of the inferior colliculus (DCIC) is distinguished from other layers by its cytoarchitecture and fiber connections. However, the information of the sound types represented in layer 1 of the DCIC remains unclear because placing electrodes on such thin structures is challenging. In this study, we utilized in vivo calcium imaging to assess auditory-evoked activities in multiple cells in layer 1 of DCIC and to characterize sound stimuli producing strong activity. Most cells examined showed strong responses to broad-band noise and low-frequency tone bursts of high sound intensity. In some cases, we successfully obtained frequency response areas, which are receptive fields to tone frequencies and intensities, and ~30% of these showed V-shape tunings. This is the first systematic study to record auditory responses of cells in layer 1 of DCIC. These results indicate that cells in this area are selective to tones with low frequency, implying the importance of such auditory information in the neural circuitry of layer 1 of DCIC.

Effectiveness of thermotherapy using a heat and steam generating sheet for cartilage in knee osteoarthritis.

[Purpose] Superficial heat treatment is one of the most widely used physical therapies for osteoarthritis (OA). We clinically evaluated the effects of local heat treatment and exercise therapy for knee OA, and evaluated the articular cartilage using magnetic resonance imaging (MRI) T2 mapping. [Subjects and Methods] Eighteen females aged 50-69 (59.5 ± 8.5 years, mean ± SD) years diagnosed with early-stage knee OA were randomly assigned using computer-generated random numbers to either a local heat treatment group (LH group, 9 subjects) or an exercise therapy group (EX group, 9 subjects). These groups were subjected to a 12-week intervention experiment. MRI T2 mapping was performed for cartilage imaging and quantitative evaluation. For clinical evaluation, the Japanese Knee Osteoarthritis Measure (JKOM) and the Timed Up and Go (TUG) test were performed. Both clinical and MRI evaluations were performed at the beginning and end of the intervention (0 week (Time 0) and 12 weeks). [Results] The total JKOM score had a significantly decreased in the LH group at 12 weeks. However, in the EX group the total JKOM scores at Time 0 and 12 weeks were not significantly different. The TUG time in the EX group was significant shorter at 12 weeks, whereas it showed no significant change in the LH group at 12 weeks, though the TUG times of 7 of the 9 patients decreased, exhibiting some improvement. The T2 value of the LH group was significantly shorter at 12 weeks. However, the T2 value in the EX group showed no significant change at 12 weeks. [Conclusion] After local heat treatment using heat- and steam moisture-generating sheets for 12 weeks, we observed improvements in clinical symptoms and walking abilities. Moreover, positive effects on cartilage metabolism were suggested.

Contribution of anterior cingulate cortex and descending pain inhibitory system to analgesic effect of lemon odor in mice.

BACKGROUND: Affections are thought to regulate pain perception through the descending pain inhibitory system in the central nervous system. In this study, we examined in mice the affective change by inhalation of the lemon oil, which is well used for aromatherapy, and the effect of lemon odor on pain sensation. We also examined the anterior cingulate cortex (ACC) and descending pain inhibitory system to such regulation of pain. RESULTS: In the elevated plus maze, the time spent in the open arms was increased by inhalation of lemon oil. The pain behavior induced by injection of formalin into the hind paw was decreased. By inhalation of lemon oil, the number of c-Fos expression by formalin injection was significantly increased in the ACC, periaqueductal grey (PAG), nucleu raphe magnus (NRM) and locus ceruleus, and decreased in the spinal dorsal horn (SDH). The destruction of the ACC with ibotenic acid led to prevent the decrease of formalin-evoked nocifensive behavior in mice exposed to lemon oil. In these mice, the change of formalin-induced c-Fos expression in the ACC, lateral PAG, NRM and SDH by lemon odor was also prevented. Antagonize of dopamine D1 receptor in the ACC prevented to the analgesic effect of lemon oil. CONCLUSIONS: These results suggest that the analgesic effect of lemon oil is induced by dopamine-related activation of ACC and the descending pain inhibitory system.

'Goose bumps'-associated anaemia.

A 36-year-old woman presented to our hospital for further evaluation of anaemia screened by regular check-up. She was diagnosed with iron-deficiency anaemia (IDA) but refractory to iron supplementation. She had negative results of occult blood in the stool and no gynaecologic disease potentially causing hyper menorrhea. Upper endoscopy revealed a nodular gastritis, which has been called 'Torihada-ien', a Japanese word meaning 'gastritis with goose bumps-like appearance', associated with Helicobacter pylori (Hp) infection. After Hp eradication therapy, her anaemia resolved with an improved response to iron supplementation. In an unexplained IDA patient, Hp-associated gastritis should be considered in the possible background.

High-dose zoledronic acid impacts bone remodeling with effects on osteoblastic lineage and bone mechanical properties.

PURPOSE: The increasing incidence of osteonecrosis of the jaw and its possible association with high cumulative doses of bisphosphonate led us to study the effects of high doses of zoledronic acid (ZA) on bone remodeling. EXPERIMENTAL DESIGN: Five-week-old C57BL6 mice were treated with saline or ZA weekly for 3 weeks at increasing doses (0.05-1 mg/Kg). Effects of ZA on bone remodeling were studied using standard assays. RESULTS: We observed an increase in bone mineral density and content in treated animals at doses of 0.05 mg/Kg, which was not further enhanced at higher doses of ZA. Trabecular bone volume at the proximal tibia and the distal femur assessed by histomorphometry and microCT, respectively, increased significantly in ZA-treated groups. There was however no difference between 0.5 and 1 mg/kg, suggesting a ceiling effect for ZA. ZA led to decreased numbers of osteoclasts and osteoblasts per bone perimeter that paralleled a significant reduction of serum levels of TRAC5b and osteocalcin in vivo. Effects on osteoblasts were confirmed in in vitro assays. Mechanical testing of the femur showed increased brittleness in ZA-treated mice. CONCLUSIONS: High doses of ZA inhibit both osteoclast and osteoblasts function and bone remodeling in vivo interfering with bone mechanical properties. No dose response was noted beyond 0.5 mg/kg suggesting that lower doses of ZA may be adequate in inhibiting bone resorption. Our data may help inform future studies of ZA use with respect to alternate and lower doses in the treatment of patients with cancer bone disease.

Effect of heat- and steam-generating sheet on daily activities of living in patients with osteoarthritis of the knee: randomized prospective study.

BACKGROUND: Thermotherapy is widely known to be effective for osteoarthritis of the knee (knee OA), but most treatment methods make use of dry heat. We developed a sheet that generates heat and steam simultaneously. In this prospective randomized study, we evaluated the effectiveness of this sheet. METHODS: Of 41 female patients with knee OA randomized to use the heat/steam-generating sheet or the dry heat-generating sheet, 37 patients (20 using the heat/steam-generating sheet and 17 using the dry heat-generating sheet) who used the sheets continuously for 4 weeks were studied. Outcome measures included the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and Japan Orthopaedic Association (JOA) scores, which were applied at baseline and after 2 and 4 weeks of use. RESULTS: Significant improvement of the total WOMAC score was observed at 2 and 4 weeks (compared to baseline) in the heat/steam-generating sheet group, but no significant change was observed in the dry heat-generating sheet group. Among the JOA scores, the gait ability score was also improved significantly only in the heat/steam-generating sheet group. The effects were still seen 6 weeks after completion of treatment. CONCLUSIONS: The present study provided evidence that the heat/steam-generating sheet that we developed is effective for alleviating pain and is especially superior in regard to improving stiffness and gait impairment in patients with knee OA. Furthermore, the effect persists for at least 6 weeks after application.

MLN3897, a novel CCR1 inhibitor, impairs osteoclastogenesis and inhibits the interaction of multiple myeloma cells and osteoclasts.

The interaction between osteoclasts (OCs) and multiple myeloma (MM) cells plays a key role in the pathogenesis of MM-related osteolytic bone disease (OBD). MM cells promote OC formation and, in turn, OCs enhance MM cell proliferation. Chemokines are mediators of MM effects on bone and vice versa; in particular, CCL3 enhances OC formation and promotes MM cell migration and survival. Here, we characterize the effects of MLN3897, a novel specific antagonist of the chemokine receptor CCR1, on both OC formation and OC-MM cell interactions. MLN3897 demonstrates significant impairment of OC formation (by 40%) and function (by 70%), associated with decreased precursor cell multinucleation and down-regulation of c-fos signaling. OCs secrete high levels of CCL3, which triggers MM cell migration; conversely, MLN3897 abrogates its effects by inhibiting Akt signaling. Moreover, MM cell-to-OC adhesion was abrogated by MLN3897, thereby inhibiting MM cell survival and proliferation. Our results therefore show novel biologic sequelae of CCL3 and its inhibition in both osteoclastogenesis and MM cell growth, providing the preclinical rationale for clinical trials of MLN3897 to treat OBD in MM.

Evaluation of morphological and molecular variation in Plantago asiatica var. densiuscula, with special reference to the systematic treatment of Plantago asiatica var. yakusimensis.

Morphological and molecular variations in Plantago asiatica L. var. densiuscula Pilg. were analyzed to evaluate the genetic basis for recognizing the dwarf variety P. asiatica var. yakusimensis (Masam.) Ohwi. Considerable variation in the leaf size of P. asiatica var. densiuscula was observed, and no morphological discontinuities were found between the dwarf types of P. asiatica var. densiuscula and P. asiatica var. yakusimensis. Morphological analysis of plants grown under standardized conditions revealed that both environmental plasticity and genetic differentiation contributed to the dwarfisms. Molecular phylogenetic analysis of rDNA internal transcribed spacer (ITS) regions and the SUC1 locus encoding a sucrose transporter revealed that P. asiatica var. yakusimensis was genetically unique although the differentiation level was low. From the above results, we concluded that P. asiatica var. yakusimensis should be reduced to a form of P. asiatica var. densiuscula. Furthermore, the geographic distribution of the SUC1 genotype suggested multiple origins of dwarves, and possible hypotheses for the origins of dwarves are discussed.

Small-field radiotherapy in combination with concomitant chemotherapy for locally advanced pancreatic carcinoma.

PURPOSE: To evaluate the effectiveness of small-field radiotherapy in combination with concomitant 5-fluorouracil (5FU) or cisplatin for locally advanced pancreatic carcinoma. MATERIALS AND METHODS: From November 1993 to January 1999, 53 patients underwent continuous 5FU infusion at 200mg/m2 (27 patients) or a 30-min cisplatin infusion at 5mg/m2/day (26 patients) just prior to each irradiation. The radiation field was limited to cover the primary and the paraaortic regions at celiac and supramesenteric axis levels. A total dose of 50.4Gy in 28 sessions was given in 5.6 weeks. RESULTS: Median and 1-year survival rates were 10.2 months and 35.2%, respectively. Local failure occurred in 19 patients (36%) and liver metastases in 16 patients (30%). All local recurrences occurred only within the radiation field. CONCLUSIONS: Median survival rates were comparable to other studies. Because local failure occurred only within the radiation field, the use of relatively small-field radiotherapy may be justified in the treatment of locally advanced pancreatic carcinoma in addition to concurrent administration of either 5FU or cisplatin.