Tetrahydrobiopterin prevents chronic ischemia-related lower urinary tract dysfunction through the maintenance of nitric oxide bioavailability.

This study aimed to investigate the influence of chronic ischemia on nitric oxide biosynthesis in the bladder and the effect of administering tetrahydrobiopterin (BH4), a cofactor for endothelial nitric oxide synthase (eNOS), on chronic ischemia-related lower urinary tract dysfunction (LUTD). This study divided male Sprague-Dawley rats into Control, chronic bladder ischemia (CBI) and CBI with oral BH4 supplementation (CBI/BH4) groups. In the CBI group, bladder capacity and bladder muscle strip contractility were significantly lower, and arterial wall was significantly thicker than in Controls. Significant improvements were seen in bladder capacity, muscle strip contractility and arterial wall thickening in the CBI/BH4 group as compared with the CBI group. Western blot analysis of bladder showed expressions of eNOS (p = 0.043), HIF-1α (p < 0.01) and dihydrofolate reductase (DHFR) (p < 0.01), which could regenerate BH4, were significantly higher in the CBI group than in Controls. In the CBI/BH4 group, HIF-1α (p = 0.012) and DHFR expressions (p = 0.018) were significantly decreased compared with the CBI group. Our results suggest that chronic ischemia increases eNOS and DHFR in the bladder to prevent atherosclerosis progression. However, DHFR could not synthesize sufficient BH4 relative to the increased eNOS, resulting in LUTD. BH4 supplementation protects lower urinary tract function by promoting eNOS activity.

Yokukansan, a Herbal Medicine in Japan, Buffers Social Crowding Stress via Ameliorating Glucocorticoid Secretion Response to Vasopressin.

On the basis of the data that yokukansan (YKS), a herbal medicine, ameliorates aggressive behavior and abnormal glucocorticoid secretion of socially isolated mice under zinc deficiency, we tested whether YKS preventively buffers crowding stress-induced attenuations of glucocorticoid secretion response and long-term potentiation (LTP), an index of cognition. YKS-containing water was administered during the period of exposure to social crowding stress for 3 weeks. Serum corticosterone level was not significantly modified by administration of YKS-containing water and was also not increased after social-crowding stress. When vasopressin was injected into crowding-stressed rats to assess corticosterone secretion via pituitary-adorenocortical axis activation, vasopressin-induced increase in serum corticosterone was significantly attenuated compared to non-stressed control rats, indicating that the pituitary-adrenocortical response to vasopressin is affected after exposure to crowding stress. Interestingly, administration of YKS-containing water rescued attenuation of vasopressin-induced increase in serum corticosterone. LTP at Schaffer collateral-CA1 pyramidal cells synapses was attenuated in the hippocampal slices from crowding-stressed rats, while administration of YKS-containing water rescued the attenuation. The present study demonstrates that intake of YKS rescues crowding stress-induced impairments of glucocorticoid secretion response to vasopressin and hippocampal LTP. The intake of YKS may be benefit to buffering chronic stress.

Oral Administration of Apple Procyanidins Ameliorates Insulin Resistance via Suppression of Pro-Inflammatory Cytokine Expression in Liver of Diabetic ob/ob Mice.

Procyanidins, the main ingredient of apple polyphenols, are known to possess antioxidative and anti-inflammatory effects associated closely with the pathophysiology of insulin resistance and type 2 diabetes. We investigated the effects of orally administered apple procyanidins (APCs) on glucose metabolism using diabetic ob/ob mice. We found no difference in body weight or body composition between mice treated with APCs and untreated mice. A 4 week oral administration of APCs containing water [0.5% (w/v)] ameliorated glucose tolerance, insulin resistance, and hepatic gluconeogenesis in ob/ob mice. APCs also suppressed the increase in the level of the pancreatic ß-cell. Insulin-stimulated Akt phosphorylation was significantly enhanced; pro-inflammatory cytokine expression levels were significantly decreased, and c-Jun N-terminal kinase phosphorylation was downregulated in the liver of those mice treated with APCs. In conclusion, APCs ameliorate insulin resistance by improving hepatic insulin signaling through suppression of hepatic inflammation in ob/ob mice, which may be a mechanism with possible beneficial health effects of APCs in disturbed glucose metabolism.

Novel stress increases hypothalamic-pituitary-adrenal activity in mice with a raised bite.

BACKGROUND AND OBJECTIVE: In humans, occlusal disharmony may cause various physical complaints, including head and neck ache, stiffness in the shoulder and neck, and arthrosis of the temporomandibular joints. Occlusal disharmony induced by raising the bite in rodents, increases plasma corticosterone levels, which leads to morphologic changes in the hippocampus and altered hippocampus-related behavior. The paraventricular nucleus (PVN) of the hypothalamus regulates the hypothalamic-pituitary-adrenal system. Chronically stressed animals exposed to a novel stress exhibit higher adrenocorticotropic hormone levels than naive control animals. We hypothesized that there would be different response of the corticotrophin releasing hormone (CRH) and arginine vasopressin (AVP) to a novel acute stress with occlusal disharmony. DESIGN: In order to investigate how exposure of mice with occlusal disharmony to a novel acute stress (restraint stress) affects the PVN, we induced occlusal disharmony by raising the vertical dimension of the bite (bite-raised condition) and examined the expression of corticotrophin releasing hormone (CRH) mRNA and arginine vasopressin (AVP) mRNA in mouse PVN. RESULTS: CRH mRNA expression was increased in the PVN of the bite-raised group 90min after the bite-raising procedure, but the expression was recovered to the control level at 14days. AVP mRNA expression in the PVN was normal at 90min, and increased significantly 14days after the bite-raising procedure. Exposure to restraint stress in the bite-raised mice induced a significant increase in CRH mRNA expression in the PVN. CONCLUSIONS: The bite-raising procedure induced a rapid CRH mRNA response and a slower AVP mRNA response in the parvocellular PVN of the hypothalamus. Exposure to a novel stress following the bite-raising procedure further reinforced the CRH stress response. Thus, occlusal disharmony, such as that induced by raising the bite, may be a risk factor for hypersensitivity to a novel stress.

Immune Milk Suppresses Herpes Simplex Virus Type 1.

Immune milk has been developed as a substitute for colostrum and contains a high concentration of IgG antibodies specific to the immunized pathogens. Meanwhile, bovine herpesvirus type-1 (BHV-1) naturally infects cattle worldwide, and its antibody is found in milk. Moreover, BHV-1 glycoprotein K, the major antigen, exhibits substantial homology with human herpes virus simplex 1 (HSV-1) glycoprotein K. On the basis of this evidence, we hypothesized BHV-1 antibody exists in immune milk and suppresses HSV-1 activity. This study investigated whether immune milk IgG recognizes HSV-1 and suppresses HSV-1 activity. IgG in immune milk was purified by affinity Protein A columns, and HSV-1-reactive IgG in immune milk IgG was detected and quantified by ELISA. The efficacy of the IgG against HSV-1 was analyzed using a reduction assay based on the cytopathic effect due to HSV-1 in the presence of macrophages. We detected a high concentration of HSV-1-reactive IgG in immune milk. Furthermore, IgG suppressed HSV-1 pathogenicity in the presence of macrophages. These results indicate immune milk has protective activity against HSV-1 by opsonic activity owing to its high concentration of HSV-1-reactive IgG, which is likely the BHV-1 antibody. HSV-1 is currently a refractory infection with a worldwide distribution. Primary infection occurs via the oral cavity, but there is no effective precaution at this time. However, the present results suggest that taking oral immune milk may be an effective measure to prevent primary HSV-1 infection in the oral cavity.

Proteomic study of sera from patients with bladder cancer: usefulness of S100A8 and S100A9 proteins.

To effectively obtain tumour-specific markers, fractionated proteins obtained using reversed-phase high-performance liquid chromatography for patient-matched pre- and postoperative sera from bladder cancer patients were compared by two-dimensional gel electrophoresis. The usefulness of the identified proteins was confirmed immunohistochemically. S100A8 and S100A9 were identified as tumour-associated proteins. The increased immunoreactive expression of S100A8 protein was associated with bladder wall muscle invasion of the tumour and cancer-specific survival (p<0.05), and the increased immunoreactive expression of S100A9 protein was associated with the tumour grade (p<0.05). In addition, increased expressions of both proteins was associated with recurrence-free survival at a median follow-up of 32.9 months (both p<0.05). On multivariate analysis, the expression of S100A8 was a significant predictor of recurrence (p<0.05). These findings may help to identify biologically aggressive tumors and, thus, patients who might benefit from more intensive adjuvant therapy.

Crystal structures of the lumazine protein from Photobacterium kishitanii in complexes with the authentic chromophore, 6,7-dimethyl- 8-(1'-D-ribityl) lumazine, and its analogues, riboflavin and flavin mononucleotide, at high resolution.

Lumazine protein (LumP) is a fluorescent accessory protein having 6,7-dimethyl-8-(1'-d-ribityl) lumazine (DMRL) as its authentic chromophore. It modulates the emission of bacterial luciferase to shorter wavelengths with increasing luminous strength. To obtain structural information on the native structure as well as the interaction with bacterial luciferase, we have determined the crystal structures of LumP from Photobacterium kishitanii in complexes with DMRL and its analogues, riboflavin (RBF) and flavin mononucleotide (FMN), at resolutions of 2.00, 1.42, and 2.00 A. LumP consists of two beta barrels that have nearly identical folds, the N-terminal and C-terminal barrels. The structures of LumP in complex with all of the chromophores studied are all essentially identical, except around the chromophores. In all of the structures, the chromophore is tethered to the narrow cavity via many hydrogen bonds in the N-terminal domain. These are absent in the C-terminal domain. Hydrogen bonding in LumP-FMN is decreased in comparison with that in LumP-RBF because the phosphate moiety of FMN protrudes out of the narrow cavity. In LumP-DMRL, the side chain of Gln65 is close to the ring system, and a new water molecule that stabilizes the ligand is observed near Ser48. Therefore, DMRL packs more tightly in the ligand-binding site than RBF or FMN. A docking simulation of bacterial luciferase and LumP suggests that the chromophore is located close enough for direct energy transfer to occur. Moreover, the surface potentials around the ligand-binding sites of LumP and bacterial luciferase exhibit complementary charge distributions, which would have a significant effect on the interaction between LumP and luciferase.

Clinical and electrophysiological characteristics of binodal disease.

BACKGROUND: Although coexistence of atrioventricular conduction disturbances with sick sinus syndrome (SSS), so-called binodal disease (BND), is a frequently encountered disorder, its clinical significance and electrophysiological characteristics remain unknown. METHODS AND RESULTS: One hundred and seven patients with SSS were divided into BND (n=30) and N-BND groups (n=77). Sinus cycle length, sinus node recovery time (SRT), sino-atrial conduction time (SACT), the number of isolated sinus node electrograms, atrio-His (AH) interval, His-ventricular (HV) interval, intra-atrial conduction time (PA intervals) and QRS width were measured. In addition, the prevalence of bundle-branch block was obtained. The parameters of sino-atrial and intra-atrial conduction were significantly longer in the BND group: SRT (5,070+/-2,628 vs 3,122+/-1,856 ms, p<0.05), SACT (115+/-30 vs 87+/-21 ms, p<0.05), PA intervals (56+/-13 vs 41+/-8 ms, p<0.05). The BND group was more likely to have atrial fibrillation than the N-BND group (83.3% vs 53.2%, p<0.01). HV interval, QRS width and the prevalence of associated bundle-branch block did not differ between the 2 groups. CONCLUSION: BND patients not only had sino-atrial and atrioventricular node dysfunction, but also widespread atrial conduction disturbances. Thus, in the clinical setting BND should be categorized as severe SSS.

[Anti-Candida albicans activity of essential oils including Lemongrass (Cymbopogon citratus) oil and its component, citral].

The effects of 12 essential oils, popularly used as antifungal treatments in aromatherapy, on growth of Candida albicans were investigated. Mycelial growth of C. albicans, which is known to give the fungus the capacity to invade mucosal tissues, was inhibited in the medium containing 100 micro g/ml of the oils: lemongrass (Cymbopogon citratus), thyme (Thymus vulgaris), patchouli (Pogostemon cablin) and cedarwood (Cedrus atlantica). Not only lemongrass oil but also citral, a major component of lemongrass oil (80%), in the range of 25 and 200 micro g/ml inhibited the mycelial growth but allowed yeast-form growth. More than 200 micro g/ml of citral clearly inhibited both mycelial and yeast-form growth of C. albicans. These results provide experimental evidence suggesting the potential value of lemongrass oil for the treatment of oral or vaginal candidiasis.