Effects of Sulfonylureas on Periodontopathic Bacteria-Induced Inflammation.

Interleukin-1ß (IL-1ß) is an inflammatory cytokine produced by monocytes/macrophages and is closely associated with periodontal diseases. The NLRP3 inflammasome is involved in IL-1ß activation through pro-IL-1ß processing and pyroptotic cell death in bacterial infection. Recently, glyburide, a hypoglycemic sulfonylurea, has been reported to reduce IL-1ß activation by suppressing activation of the NLRP3 inflammasome. Therefore, we evaluated the possibility of targeting the NLRP3 inflammasome pathway by glyburide to suppress periodontal pathogen-induced inflammation. THP-1 cells (a human monocyte cell line) were differentiated to macrophage-like cells by treatment with phorbol 12-myristate 13-acetate and stimulated by periodontopathic bacteria, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, or Fusobacterium nucleatum, in the presence of glyburide. IL-1ß and caspase-1 expression in the cells and culture supernatants were analyzed by Western blotting and enzyme-linked immunosorbent assay, and cell death was analyzed by lactate dehydrogenase assay. Stimulation of THP-1 macrophage-like cells with every periodontopathic bacteria induced IL-1ß secretion without cell death, which was suppressed by the NLRP3 inhibitor, MCC950, and caspase-1 inhibitor, z-YVAD-FMK. Glyburide treatment suppressed IL-1ß expression in culture supernatants and enhanced intracellular IL-1ß expression, suggesting that glyburide may have inhibited IL-1ß secretion. Subsequently, a periodontitis rat model was generated by injecting periodontal bacteria into the gingiva, which was analyzed histologically. Oral administration of glyburide significantly suppressed the infiltration of inflammatory cells and the number of osteoclasts in the alveolar bone compared with the control. In addition to glyburide, glimepiride was shown to suppress the release of IL-1ß from THP-1 macrophage-like cells, whereas other sulfonylureas (tolbutamide and gliclazide) or other hypoglycemic drugs belonging to the biguanide family, such as metformin, failed to suppress IL-1ß release. Our results suggest that pharmacological targeting of the NLRP3 pathway may be a strategy for suppressing periodontal diseases.

Whole-Genome Sequencing of the NARO World Rice Core Collection (WRC) as the Basis for Diversity and Association Studies.

Genebanks provide access to diverse materials for crop improvement. To utilize and evaluate them effectively, core collections, such as the World Rice Core Collection (WRC) in the Genebank at the National Agriculture and Food Research Organization, have been developed. Because the WRC consists of 69 accessions with a high degree of genetic diversity, it has been used for >300 projects. To allow deeper investigation of existing WRC data and to further promote research using Genebank rice accessions, we performed whole-genome resequencing of these 69 accessions, examining their sequence variation by mapping against the Oryza sativa ssp. japonica Nipponbare genome. We obtained a total of 2,805,329 single nucleotide polymorphisms (SNPs) and 357,639 insertion-deletions. Based on the principal component analysis and population structure analysis of these data, the WRC can be classified into three major groups. We applied TASUKE, a multiple genome browser to visualize the different WRC genome sequences, and classified haplotype groups of genes affecting seed characteristics and heading date. TASUKE thus provides access to WRC genotypes as a tool for reverse genetics. We examined the suitability of the compact WRC population for genome-wide association studies (GWASs). Heading date, affected by a large number of quantitative trait loci (QTLs), was not associated with known genes, but several seed-related phenotypes were associated with known genes. Thus, for QTLs of strong effect, the compact WRC performed well in GWAS. This information enables us to understand genetic diversity in 37,000 rice accessions maintained in the Genebank and to find genes associated with different phenotypes. The sequence data have been deposited in DNA Data Bank of Japan Sequence Read Archive (DRA) (Supplementary Table S1).

p11 in Cholinergic Interneurons of the Nucleus Accumbens Is Essential for Dopamine Responses to Rewarding Stimuli.

A recent study showed that p11 expressed in cholinergic interneurons (CINs) of the nucleus accumbens (NAc) is a key regulator of depression-like behaviors. Dopaminergic neurons projecting to the NAc are responsible for reward-related behaviors, and their function is impaired in depression. The present study investigated the role of p11 in NAc CINs in dopamine responses to rewarding stimuli. The extracellular dopamine and acetylcholine (ACh) levels in the NAc were determined in freely moving male mice using in vivo microdialysis. Rewarding stimuli (cocaine, palatable food, and female mouse encounter) induced an increase in dopamine efflux in the NAc of wild-type (WT) mice. The dopamine responses were attenuated (cocaine) or abolished (food and female mouse encounter) in constitutive p11 knock-out (KO) mice. The dopamine response to cocaine was accompanied by an increase in ACh NAc efflux, whereas the attenuated dopamine response to cocaine in p11 KO mice was restored by activation of nicotinic or muscarinic ACh receptors in the NAc. Dopamine responses to rewarding stimuli and ACh release in the NAc were attenuated in mice with deletion of p11 from cholinergic neurons (ChAT-p11 cKO mice), whereas gene delivery of p11 to CINs restored the dopamine responses. Furthermore, chemogenetic studies revealed that p11 is required for activation of CINs in response to rewarding stimuli. Thus, p11 in NAc CINs plays a critical role in activating these neurons to mediate dopamine responses to rewarding stimuli. The dysregulation of mesolimbic dopamine system by dysfunction of p11 in NAc CINs may be involved in pathogenesis of depressive states.

Translocator protein 18kDa antagonist ameliorates stress-induced stool abnormality and abdominal pain in rodent stress models.

BACKGROUND: Irritable bowel syndrome (IBS) is a functional gastrointestinal (GI) disorder characterized by abdominal pain and abnormal bowel habits, both of which are exacerbated by psychological stress. The translocator protein 18kDa (TSPO) is a marker of reactive gliosis in a number of central nervous system (CNS) diseases and responsible for many cellular functions, including neurosteroidogenesis. Although it has been reported that psychological stress disturbs neurosteroids levels, the pathophysiological relevance of TSPO in IBS is poorly understood. METHODS: We examined the effects of a TSPO antagonist, ONO-2952, on stress-induced stool abnormality and abdominal pain in rats, and on anxiety-related behavior induced by cholecystokinin. KEY RESULTS: Oral administration of ONO-2952 attenuated stress-induced defecation and rectal hyperalgesia in rats with an efficacy equivalent to that of a 5-HT3 receptor antagonist. In addition, ONO-2952 suppressed cholecystokinin-induced anxiety-like behavior with an efficacy equivalent to that of psychotropic drugs. On the other hand, ONO-2952 did not affect spontaneous defecation, gastrointestinal transit, visceral nociceptive threshold, and neurosteroid production in non-stressed rats even at a dose 10 times higher than its effective dose in the stress models. CONCLUSIONS AND INFERENCES: These results suggest that TSPO antagonism results in antistress action, and that ONO-2952 is a promising candidate for IBS without side effects associated with current treatment.

Proton pump inhibitor after endoscopic resection for esophageal squamous cell cancer: multicenter prospective randomized controlled trial.

BACKGROUND: Whether proton pump inhibitors (PPIs) relieve heartburn or precordial pain after endoscopic resection (ER) for esophageal squamous cell carcinoma (ESCC) remains unclear. The aim of this study was to investigate the efficacy of PPI therapy for these symptoms after ER for ESCC. METHODS: We conducted a multicenter prospective randomized controlled trial among 15 hospitals in Japan. In total, 229 patients with cT1a ESCC were randomly assigned to receive PPI therapy for 5 weeks after ER (the PPI group, n = 115) or follow-up without PPI therapy (the non-PPI group, n = 114). The primary end point was the incidence of gastroesophageal reflux disease (GERD)-like symptoms after ER from a self-reported questionnaire (Frequency Scale for Symptoms of GERD). Secondary end points were ulcer healing rate at 5 weeks, incidence of pain, improvement rate of symptoms in those who started PPI therapy because of GERD-like symptoms in the non-PPI group, and adverse events. RESULTS: No significant difference was observed in the incidence of GERD-like symptoms after ER between the non-PPI and PPI groups (30 % vs 34 %, respectively). No significant differences were observed in the ulcer healing rate at 5 weeks (84 % vs 85 %) and incidence of pain within 1 week (36 % vs 45 %). In nine of ten patients (90 %) who started PPI therapy because of GERD-like symptoms in the non-PPI group, PPI administration relieved GERD-like symptoms. No adverse events related to PPI administration were observed. CONCLUSION: PPI therapy is not efficacious in reducing symptoms and did not promote healing of ulcers in patients undergoing ER for ESCC.

Role of macrophage colony-stimulating factor (M-CSF)-dependent macrophages in gastric ulcer healing in mice.

We examined the role of macrophage colony-stimulating factor (M-CSF)-dependent macrophages in the healing of gastric ulcers in mice. Male M-CSF-deficient (op/op) and M-CSF-expressing heterozygote (+/?) mice were used. Gastric ulcers were induced by thermal cauterization under ether anesthesia, and healing was observed for 14 days after ulceration. The numbers of macrophages and microvessels in the gastric mucosa were determined immunohistochemically with anti-CD68 and anti-CD31 antibodies, respectively. Expression of tumor necrosis factor (TNF)-α, cyclooxygenase (COX)-2, and vascular endothelial growth factor (VEGF) mRNA was determined via real-time reverse transcription-polymerase chain reaction (RT-PCR), and the mucosal content of prostaglandin (PG) E(2) was determined via enzyme immunoassay on day 10 after ulceration. The healing of gastric ulcers was significantly delayed in op/op mice compared with +/? mice. Further, significantly fewer macrophages were observed in the normal gastric mucosa of op/op mice than in +/? mice. Ulcer induction caused a marked accumulation of macrophages around the ulcer base in +/? mice, but this response was attenuated in op/op mice. The mucosal PGE(2) content as well as the expression of COX-2, VEGF, and TNF-α mRNA were all upregulated in the ulcerated area of +/? mice but significantly suppressed in op/op mice. The degree of vascularization in the ulcerated area was significantly lower in op/op mice than in +/? mice. Taken together, these results suggest that M-CSF-dependent macrophages play an important role in the healing of gastric ulcers, and that this action may be associated with angiogenesis promoted by upregulation of COX-2/PGE(2) production.

[Surgical treatment of postinfarction ventricular septal rupture].

BACKGROUND: Postinfarction ventricular septal rupture (VSR) is a lethal complication with high mortality. The aim of this study was to evaluate our surgical strategy and results of VSR. PATIENTS AND METHODS: Between 1996 and 2008, 13 consecutive patients underwent operation for VSR at our hospital. All patients required emergent operation because of severe cardiogenic shock. Surgical procedure consisted of endocardial patch repair with infarct exclusion, so called "Komeda-David operation". In patients with multiple coronary artery disease, myocardial revascularization was done simultaneously. RESULTS: These patients were divided into 2 groups according to the location of VSR. There were 9 patients of anterior VSR. Two of them could not be weaned from cardiopulmonary bypass and died of severe low output syndrome (LOS) at early postoperative period. The site of infarction in both patients was broad anteroseptal region including right ventricle. On the other hand, there were 4 patients of inferior VSP. Two of these patients were lost due to LOS. One patient was complicated with left ventricular free wall rupture. In another patient, infarction was extended proximally toward the mitral annulus and papillary muscles. Both cardiopulmonary bypass time and aortic crossclamp time were significantly longer in inferior VSR than in anterior region. There was no late death in 2 groups. CONCLUSIONS: Despite improvements of surgical procedures, such as infarct exclusion technique, the operative mortality remains high in cases with broad infarction and/or right ventricular infarction. In these particular circumstances, in should be mandatory to consider the optimal timing of operation and the modification of surgical technique itself.

Prospective randomized controlled study on the effects of perioperative administration of a neutrophil elastase inhibitor to patients undergoing video-assisted thoracoscopic surgery for thoracic esophageal cancer.

Sivelestat sodium hydrate (Ono Pharmaceutical Co., Osaka, Japan) is a selective inhibitor of neutrophil elastase (NE) and is effective in reducing acute lung injury associated with systemic inflammatory response syndrome (SIRS). We conducted a prospective randomized controlled study to investigate the efficacy of perioperative administration of sivelestat sodium hydrate to prevent postoperative acute lung injury in patients undergoing thoracoscopic esophagectomy and radical lymphadenectomy. Twenty-two patients with thoracic esophageal cancer underwent video-assisted thoracoscopic esophagectomy with extended lymph node dissection in our institution between April 2007 and November 2008. Using a double-blinded method, these patients were randomly assigned to one of two groups preoperatively. The active treatment group received sivelestat sodium hydrate intravenously for 72 hours starting at the beginning of surgery (sivelestat-treated group; n= 11), while the other group received saline (control group; n= 11). All patients were given methylprednisolone immediately before surgery. Postoperative clinical course was compared between the two groups. Two patients (one in each group) were discontinued from the study during the postoperative period because of surgery-related complications. Of the remaining 20 patients, 2 patients who developed pneumonia within a week after surgery were excluded from some laboratory analyses, so data from 18 patients (9 patients in each group) were analyzed based on the arterial oxygen pressure/fraction of inspired oxygen ratio, white blood cell count, serum C-reactive protein level, plasma cytokine levels, plasma NE level, and markers of alveolar type II epithelial cells. In the current study, the incidence of postoperative morbidity did not differ between the two groups. The median duration of SIRS in the sivelestat-treated group was significantly shorter than that in the control group: 17 (range 9-36) hours versus 49 (15-60) hours, respectively (P= 0.009). Concerning the parameters used for the diagnosis of SIRS, the median heart rates on postoperative day (POD) 2 were significantly lower in the sivelestat-treated group than in the control group (P= 0.007). The median arterial oxygen pressure/fraction of inspired oxygen ratio of the sivelestat-treated group were significantly higher than those of the control group on POD 1 and POD 7 (POD 1: 372.0 [range 284.0-475.0] vs 322.5 [243.5-380.0], respectively, P= 0.040; POD 7: 377.2 [339.5-430.0] vs 357.6 [240.0-392.8], P= 0.031). Postoperative white blood cell counts, serum C-reactive protein levels, plasma interleukin-1beta, tumor necrosis factor-alpha levels, and plasma NE levels did not differ significantly between the two groups at any point during the postoperative course, nor did serum Krebs von den Lungen 6, surfactant protein-A, or surfactant protein-D levels, which were used as markers of alveolar type II epithelial cells to evaluate the severity of lung injury. Plasma interleukin-8 levels were significantly lower in the sivelestat-treated group than in the control group on POD 3 (P= 0.040). In conclusion, perioperative administration of sivelestat sodium hydrate (starting at the beginning of surgery) mitigated postoperative hypoxia, partially suppressed postoperative hypercytokinemia, shortened the duration of SIRS, and stabilized postoperative circulatory status after thoracoscopic esophagectomy.

Spectral broadening of mid-infrared femtosecond pulses in GaAs.

We achieved efficient spectral broadening for mid-IR pulses of few-microjoule energy. The spectral bandwidth of the femtosecond pulses at the center wavelength of 5000 nm increased from 540 nm to 2060 nm (from 220 to 910 cm(-1) in frequency) by nonlinear propagation in a gallium arsenide single crystal. The spectral broadening was accompanied by nonlinear absorption loss of 25%. The demonstrated scheme should be available at any operation wavelength within the material transparency range and provides a useful tool in nonlinear vibrational spectroscopy.

[Surgical strategy for aortic root reconstruction in patients with Stanford type A acute aortic dissection].

BACKGROUND: Although the aortic root dissection is a common finding in patients with type A acute aortic dissection, it is potentially fatal. Several procedures are available for reconstruction of the aortic root, and the choice depends on the mode of dissection; rupture, coronary ostial disruption or pre-existing cardiovascular diseases (annulo-aortic ectasia, aortic regurgitation: AR). The purpose of this study was to evaluate our surgical strategy of type A acute aortic dissection with proximal involvement. METHODS: Between 1996 and 2008, 100 patients with type A acute aortic dissection underwent emergency operation at our hospital. Fifteen of them received aortic root intervention simultaneously. RESULTS: In the initial 3 patients, the dissected aortic root was reinforced by fibrin glue and concomitant coronary bypass grafting was performed. During the follow-up term, residual aortic insufficiency was noted in 2 patients. Aortic root reconstruction was performed in 12 patients. Of these, the most recent 2 young patients underwent a new type of aortic valve sparing procedure, so called "partial remodeling". The postoperative echocardiography demonstrated no AR. There was no operative death in this series, and the long-term results were good. CONCLUSION: The excellent outcome was demonstrated with the change of surgical strategy. Special care should be taken for precise recognition of the proximal extension of the aortic dissection at operation. Although we recommend consideration of the partial remodeling procedure as an alternative, close observation is mandatory in this particular circumstance.

[Konno procedure for congenital aortic stenosis associated with complex left ventricular outflow tract obstruction].

Two successful cases of Konno procedure for congenital aortic stenosis and left ventricular outflow tract obstruction (LVOTO) were reported herein. A 3-year-old child previously underwent definitive repair of complete atrioventricular septal defect. Follow-up echocardiography revealed progression of valvular aortic stenosis and subaortic tunnel stenosis. Second patient was a 30-year-old male with congenital aortic stenosis, severe LVOTO and funnel chest. Both patients underwent Konno procedure, and their postoperative courses were uneventful. The Konno procedure is effective and stenotic lesion could be enlarged sufficiently even in complex LVOTO. Especially in the patient of advanced age, care should be taken to fragility of the left ventricular muscle and coronary malperfusion caused by the procedure itself.

Peripherally administered ghrelin induces bimodal effects on the mesolimbic dopamine system depending on food-consumptive states.

Ghrelin induces orexigenic behavior by activation of growth hormone secretagogue 1 receptors (GHSRs) in the ventral tegmental area (VTA) as well as hypothalamus, suggesting the involvement of mesolimbic dopamine system in the action of ghrelin. The present study aimed to identify neuronal mechanisms by which peripherally administered ghrelin regulates the mesolimbic dopamine system under different food-consumptive states. Ghrelin was administered to rats peripherally (3 nmol, i.v.) as well as locally into the VTA (0.3 nmol). Dopamine in the nucleus accumbens shell (NAc) was measured by microdialysis. Peripheral administration of ghrelin decreased dopamine levels in the NAc when food was removed following ghrelin administration. This inhibitory effect was mediated through GABA(A) and N-methyl-D-aspartate (NMDA) receptors in the VTA. In contrast, when animals consumed food following ghrelin administration, dopamine levels increased robustly. This stimulatory effect was mediated through NMDA receptors, but not through GABA(A) receptors, in the VTA. Importantly, both the inhibitory and stimulatory effects of ghrelin primarily required activation of GHSRs in the VTA. Furthermore, local injection of ghrelin into the VTA induced dopamine release in the NAc and food consumption, supporting the local action of ghrelin in the VTA. In conclusion, peripherally administered ghrelin activates GHSRs in the VTA, and induces bimodal effects on mesolimbic dopamine neurotransmission depending on food-consumptive states.

Influence of Helicobacter pylori eradication on reflux esophagitis in Japanese patients.

The relationship between Helicobacter pylori eradication and reflux esophagitis is controversial. We analyzed the development of reflux esophagitis and the change in the grade of pre-existing reflux esophagitis after eradication. Enrolled were 559 Japanese patients who received eradication therapy for H. pylori. The grade of reflux esophagitis by endoscopy before and after therapy was evaluated retrospectively. No esophagitis was present before eradication in 526 patients. H. pylori was and was not eradicated in 429 and 97, respectively. Reflux esophagitis developed in 40 of the eradication group and in three of the treatment failure group, with prevalence higher with successful eradication (P = 0.04). Successful eradication and hiatus hernia were significant risk factors for reflux esophagitis development. Twenty-seven of 33 patients with pre-existing reflux esophagitis had successful eradication and six treatment failure. The reflux esophagitis grade worsened in two (Los Angeles classification from A to B) and improved in 14 patients after eradication. With treatment failure, reflux esophagitis worsened in none and improved in three patients. There showed no significant change in the grade of pre-existing reflux esophagitis after H. pylori eradication but the sample size was too small to evaluate the difference. In conclusion, the eradication of H. pylori increases the prevalence of reflux esophagitis, and hiatus hernia was a significant risk factor for the development of reflux esophagitis.

[Postoperative management using ultra-short-acting beta-blocker (landiolol) in patients with aortic stenosis after aortic valve replacement].

Paroxysmal atrial fibrillation and other types of tachyarrhythmia are common, but might be potentially fatal complications in patients undergoing aortic valve replacement due to aortic stenosis. We report the effects of ultra-short-acting beta-blocker (landiolol) in treating 2 patients with postoperative tachycardia. The heart rate was controlled adequately without subsequent hypotension and postoperative course was uneventful. This strategy seems to be safe and beneficial in this particular circumstance.

Satisfaction with bispectral index monitoring of propofol-mediated sedation during endoscopic submucosal dissection: a prospective, randomized study.

BACKGROUND AND STUDY AIMS: Endoscopic submucosal dissection (ESD) is one of the most complex and lengthy endoscopic procedures, so deep sedation during ESD is indispensable. Our study aims were to determine whether bispectral index (BIS) monitoring is useful in titrating and reducing the dose of the sedative propofol during ESD, and to measure the satisfaction of patients and endoscopists involved in this complex and lengthy endoscopic therapy. PATIENTS AND METHODS: We performed a prospective, randomized clinical trial from July 2006 to February 2008. A total of 156 patients, with gastric neoplasm to be treated using ESD, were randomized to two groups. The BIS group (n = 78) was monitored for propofol sedation using BIS, and the no-BIS group (n = 78) was monitored by standard methods only. The two groups were compared by evaluating the doses of propofol administered to patients and the satisfaction scores (scale of 0 - 10) of patients and endoscopists. RESULTS: Although there were no significant differences between the two groups in the mean dose of propofol used (BIS group vs. no-BIS group, 5.32 mg/kg/hour vs. 4.85 mg/kg/hour; P = 0.10), the satisfaction scores of the patients (9.15 vs. 7.94; P < 0.01) and endoscopists (8.53 vs. 6.42; P < 0.001) were significantly higher with BIS monitoring. CONCLUSIONS: Monitoring with BIS during the ESD procedure did not lead to a reduction in the dose of propofol required, but did lead to higher satisfaction scores from the patients and endoscopists. A complicated and prolonged endoscopic treatment such as ESD can be carried out with optimal safety, control, and comfort by using BIS to monitor propofol sedation.

[Does transradial cardiac catheterization affect coronary artery bypass grafting?].

Recent technological advances have enabled the miniaturization of catheters for coronary angiography and intervention. As a result of this advancement, the transradial approach is becoming more popular. The advantages of this approach include a lower incidence of access site complications, earlier patient ambulation, improved patient satisfaction, and lower cost. The cardiologists of our institute have introduced this technique without delay and have taken the initiative in Japan. However, there are concerns regarding the effect of transradial cardiac catheterization on the condition of radial artery grafts for coronary artery bypass grafting (CABG). In this study, we evaluated the influence of transradial catheterization on CABG. We retrospectively evaluated 157 patients who had undergone CABG using the radial artery. The condition of the grafts was assessed intraoperatively. Postoperative coronary angiography was carried out 3 months after the surgical intervention. The patency of the grafts was assessed by 2 cardiologists. One-quarter of the radial artery grafts were affected by transradial catheterization. Since most of them were located only at the puncture site, the graft itself was capable of being used for grafting after the resection of its affected distal end. The patency rate was not affected by transradial catheterization.

Relationship between muscle oxygenation kinetics and the rate of decline in peak torque during isokinetic knee extension in acute hypoxia and normoxia.

To investigate whether low FiO2 affects muscle oxygenation and the rate of decline in peak torque (DR) during isokinetic knee extension, subjects performed 50 isokinetic knee extensions at 180 degrees /s and at 0.5 Hz while inhaling low O2 gas (12 %O2; H) or air (N). Muscle oxygenation kinetics was assessed by near-infrared spectroscopy, and whole-body V.O2 and HR were measured. We calculated total-, oxy- and deoxy-hemoglobin/myoglobin concentrations (TotalHb/Mb, OxyHb/Mb, DeoxyHb/Mb), and the slopes of the change in OxyHb/Mb during exercise. SpO2 decreased in H while DR and V.O2 did not differ between the conditions. During exercise, OxyHb/Mb was lower in H than in N, and DeoxyHb/Mb was higher in H than in N. TotalHb/Mb began to increase from the resting level earlier in H. HR was higher during the latter half of the exercise in H. The slopes of the change in OxyHb/Mb were the same in the two conditions. Our results show that low FiO2 decreases SpO2 and muscle oxygenation during maximal isokinetic knee extension. However, low SpO2 and muscle oxygenation did not affect the rates of decline of peak torque. These results suggest that the decline in peak torque occurs for reasons other than O2 availability.