Increased ghrelin signaling prolongs survival in mouse models of human aging through activation of sirtuin1.

Caloric restriction (CR) is known to retard aging and delay functional decline as well as the onset of diseases in most organisms. Ghrelin is secreted from the stomach in response to CR and regulates energy metabolism. We hypothesized that in CR ghrelin has a role in protecting aging-related diseases. We examined the physiological mechanisms underlying the ghrelin system during the aging process in three mouse strains with different genetic and biochemical backgrounds as animal models of accelerated or normal human aging. The elevated plasma ghrelin concentration was observed in both klotho-deficient and senescence-accelerated mouse prone/8 (SAMP8) mice. Ghrelin treatment failed to stimulate appetite and prolong survival in klotho-deficient mice, suggesting the existence of ghrelin resistance in the process of aging. However, ghrelin antagonist hastened death and ghrelin signaling potentiators rikkunshito and atractylodin ameliorated several age-related diseases with decreased microglial activation in the brain and prolonged survival in klotho-deficient, SAMP8 and aged ICR mice. In vitro experiments, the elevated sirtuin1 (SIRT1) activity and protein expression through the cAMP-CREB pathway was observed after ghrelin and ghrelin potentiator treatment in ghrelin receptor 1a-expressing cells and human umbilical vein endothelial cells. Furthermore, rikkunshito increased hypothalamic SIRT1 activity and SIRT1 protein expression of the heart in the all three mouse models of aging. Pericarditis, myocardial calcification and atrophy of myocardial and muscle fiber were improved by treatment with rikkunshito. Ghrelin signaling may represent one of the mechanisms activated by CR, and potentiating ghrelin signaling may be useful to extend health and lifespan.

Rikkunshito induces gastric relaxation via the ß-adrenergic pathway in Suncus murinus.

BACKGROUND: Rikkunshito (RKT) is a gastroprotective herbal medicine. In this study, we investigated the role of RKT in the relaxation of the gastric body (fundus and corpus) and antrum. METHODS: We used Suncus murinus, a unique small model animal with similar gastrointestinal motility to humans and dogs. RKT was added at 0.1, 1.0, and 5.0 mg/mL to induce relaxation in vitro; the outcome measure was the intensity of relaxation. The number of spontaneous antral contractions in the absence or the presence of RKT was also counted. KEY RESULTS: Rikkunshito induced the relaxation of the gastric body and antrum and decreased the number of spontaneous antral contractions in a dose-dependent manner. The responses to RKT (1.0 mg/mL) were not affected by pretreatment with atropine, N-nitro-l-arginine methyl ester, ritanserin, or ondansetron. On the other hand, timolol almost completely reversed the relaxation induced by RKT (1.0 mg/mL) on the gastric body and antrum and the occurrence of the spontaneous antral contractions. Both butoxamine, a ß(2) -adrenoreceptor antagonist, and L 748337, a ß(3) -adrenoreceptor antagonist, but not CGP 20712, a ß(1) -adrenoreceptor antagonist, significantly reversed the RKT-induced (1.0 mg/mL) gastric relaxation. CONCLUSIONS & INFERENCES: These results indicate that RKT stimulates and modulates gastric relaxation through ß(2) - and ß(3) -adrenergic, but not ß(1) -adrenergic, pathways in S. murinus.

Anatomy and physiology of hamstring injury.

The hamstring muscles were analyzed anatomically and physiologically to clarify the specific reasons for the incidence of muscle strain of the hamstrings. For the anatomical study, hamstring muscles of 13 embalmed cadavers were dissected. For the physiological study, the knee flexor torque and surface electromyographic (EMG) signals were measured during isometric contraction of hamstring muscles in 10 healthy adults. The biceps femoris muscle long head (BF-L) and semimembranosus muscle (SM) had hemi-pennate architecture and their fiber length per total muscle length (FL/TML) was smaller than that of semtendinosus muscle (ST) and biceps femoris muscle short head (BF-S) with other architecture. The decrease of total muscle length per fiber length (ΔTML/FL) was larger in BF-L and SM than in ST and BF-S. The EMG activities at 0° of knee angle were at maximal compared with other knee angles and were of similar level in BF-L, in SM and in ST, whereas they were considerably smaller in BF-S. The EMG at 0° of knee angle activity per physiological cross-sectional area (PCSA) was about 1.6 times greater in BF-L than in SM. These results indicate the highest risk of muscle strain was in BF-L followed by SM.

Potentiation of ghrelin signaling attenuates cancer anorexia-cachexia and prolongs survival.

Cancer anorexia-cachexia syndrome is characterized by decreased food intake, weight loss, muscle tissue wasting and psychological distress, and this syndrome is a major source of increased morbidity and mortality in cancer patients. This study aimed to clarify the gut-brain peptides involved in the pathogenesis of the syndrome and determine effective treatment for cancer anorexia-cachexia. We show that both ghrelin insufficiency and resistance were observed in tumor-bearing rats. Corticotropin-releasing factor (CRF) decreased the plasma level of acyl ghrelin, and its receptor antagonist, α-helical CRF, increased food intake of these rats. The serotonin 2c receptor (5-HT2cR) antagonist SB242084 decreased hypothalamic CRF level and improved anorexia, gastrointestinal (GI) dysmotility and body weight loss. The ghrelin receptor antagonist (D-Lys3)-GHRP-6 worsened anorexia and hastened death in tumor-bearing rats. Ghrelin attenuated anorexia-cachexia in the short term, but failed to prolong survival, as did SB242084 administration. In addition, the herbal medicine rikkunshito improved anorexia, GI dysmotility, muscle wasting, and anxiety-related behavior and prolonged survival in animals and patients with cancer. The appetite-stimulating effect of rikkunshito was blocked by (D-Lys3)-GHRP-6. Active components of rikkunshito, hesperidin and atractylodin, potentiated ghrelin secretion and receptor signaling, respectively, and atractylodin prolonged survival in tumor-bearing rats. Our study demonstrates that the integrated mechanism underlying cancer anorexia-cachexia involves lowered ghrelin signaling due to excessive hypothalamic interactions of 5-HT with CRF through the 5-HT2cR. Potentiation of ghrelin receptor signaling may be an attractive treatment for anorexia, muscle wasting and prolong survival in patients with cancer anorexia-cachexia.

I.V. infusion of a drag-reducing polymer extracted from aloe vera prolonged survival time in a rat model of acute myocardial ischaemia.

BACKGROUND: I.V. infusion of drag-reducing polymers (DRPs) has been shown to improve survival time in animals subjected to haemorrhagic shock. We hypothesized that DRPs might prolong survival time in rats following acute myocardial ischaemia (AMI). METHODS: Sixteen adult male rats were anaesthetized and mechanically ventilated. An i.v. infusion of either Dextran-40 2.5% (Control, n=8) or Dextran-40 2.5% containing 50 microg ml(-1) of an aloe vera-based DRP (DRP, n=8) was initiated at 3.5 ml h(-1). The left anterior descending coronary artery was ligated. Blood pressure, skin-tissue perfusion, and heart rate were monitored and arterial blood samples were analysed. RESULTS: The mortality at 60 min following coronary ligation was 0% in the DRP group vs 50% in the control group (P=0.025). DRP-treated animals maintained higher mean arterial pressure [60.9 (5.1) vs 47.5 (5.1) mm Hg, P=0.004] and tissue perfusion [4.2 (3.4) vs 1.2 (0.5) TPU, P=0.029]. The DRP group trended towards better acid-base status with base excess [-5.0 (1.7) vs -8.1 (5.1) mmol litre(-1), P=0.083] and pH [7.42 (0.07) vs 7.35 (0.02), P=0.03]. CONCLUSIONS: Administration of nanomolar concentrations of aloe vera-based DRP prolonged survival time in animals with AMI. DRPs may offer a novel method to treat organ/tissue hypoperfusion.

A low-molecular-weight fraction of bovine colostrum and milk enhances the oxidative burst activity of polymorphonuclear leukocytes.

Bovine colostrum and milk contain many immunomodulatory components. The low-molecular-weight fraction (< 10 kDa) was separated from colostrum and milk by gel filtration chromatography, and its effect on the oxidative burst of bovine polymorphonuclear leukocytes (PMNL) was investigated in vitro. The oxidative burst activity induced by Staphylococcus aureus was considerably enhanced when PMNLs were incubated with this low-molecular-weight fraction. However, phorbol 12-myristate 13-acetate did not trigger a burst after priming with this fraction. The oxidative burst activity enhanced by this fraction was reduced after heating. These results confirmed that a low-molecular-weight substance(s) of less than 10 kDa, present in bovine milk and colostrum, enhances the oxidative burst activity of PMNL.

Immunohistochemical analyses of thyroid-specific enhancer-binding protein in the fetal and adult rat hypothalami and pituitary glands.

Thyroid-specific enhancer-binding protein (T/EBP), also known as NKX2.1 or TTF-1, regulates the expression of thyroid- and lung-specific genes. The t/ebp/Nkx2.1-null mutant mouse was stillborn but lacked the thyroid gland, pituitary gland, ventral region of the forebrain and normal lungs. These data demonstrated that T/EBP/NKX2.1 plays an important role not only in tissue-specific gene expressions in adults but also in genesis of these organs during development. Although the expression of t/ebp/Nkx2.1 in the brain has been reported, its function in the brain remains unknown. The present study was designed to determine the localization of T/EBP/NKX2.1 in the hypothalamus and pituitary gland of fetal and adult rats by immunohistochemistry as the first step toward understanding the function of T/EBP/NKX2.1 in the rat brain. In the fetal rat hypothalamus, T/EBP/NKX2.1 was localized widely in the ventral hypothalamic areas. In the adult rat brain, T/EBP/NKX2.1 was localized in the ventromedial hypothalamic nucleus, medial tuberal nucleus, arcuate nucleus and mammillary body. No T/EBP/NKX2.1 immunoreactivity was observed in the anterior or intermediate lobe of the pituitary gland in either fetal or adult rats. On the other hand, immunoreactive T/EBP/NKX2.1 was found in the posterior lobe of the pituitary gland. This paper presents results of detailed analyses of the distributions of T/EBP/NKX2.1 protein in the fetal and adult rat hypothalami and pituitary glands, and these results should provide important information for understanding the function of T/EBP/NKX2.1 in the brain.

Analysis of survivorship after total hip arthroplasty using a ceramic head.

To determine the clinical efficacy of an alumina ceramic head, 119 cemented total hip arthroplasties in 97 patients using an alumina head coupled with ethylene oxide gas sterilized polyethylene were reviewed. Eighty-two patients (101 hips) with greater than 10 years followup were evaluated clinically and radiographically (range, 10-17.6 years), and 97 patients (119 hips) were evaluated for survivorship analysis (range, 0.6-17.6 years). The average functional hip scores according to Merle d'Aubigné and Postel improved from 8.6 preoperatively to 15.0 at the final followup, and 57 patients (64 hips) had no pain. The average polyethylene wear rate was 0.15 mm/year (range, 0.04-0.34 mm/year). Patients with polyethylene wear greater than 3 mm showed significantly higher rates of acetabular loosening. Fifteen-year survival rates (with 95% confidence intervals) with radiographic evidence of aseptic loosening as the end point were 46.8% +/- 13.4% in acetabular components and 91.9% +/- 6.6% in femoral components. Fifteen-year survival rates of hip arthroplasties with revision because of aseptic loosening as the end point were 75.3% +/- 10.2% and 97.9% +/- 3.0%, respectively. Results of the current study suggest that using an alumina head instead of a metal head may not be beneficial when coupled with ethylene oxide gas sterilized polyethylene.

Lack of noninvasive markers of ventricular repolarization inhomogeneity in patients with idiopathic ventricular tachyarrhythmia.

Noninvasive markers reflecting repolarization inhomogeneity have been proposed to be useful indices for identifying patients at risk of ventricular arrhythmias based on organic heart disease. In this study, we clarify whether or not repolarization inhomogeneity markers are useful in patients with idiopathic ventricular tachycardia (VT). We investigated T-wave alternans (TWA) and corrected QT-interval dispersion (QTD) in 84 consecutive patients with idiopathic VT, 90 patients with VT associated with organic heart disease (organic VT), and 87 normal individuals. VT was defined as tachycardia lasting > or =5 consecutive ventricular ectopic beats at a rate of > or =120 beats/min. TWA was positive in 20 of 84 patients (24%) with idiopathic VT, 59 of 90 patients (66%) with organic VT, and 16 of 87 normal individuals (18%). The alternans voltage was 2.6 +/- 3.1 micro V in idiopathic VT patients, 5.6 +/- 6.4 micro V in organic VT patients, and 2.9 +/- 5.7 micro V in normal individuals. QTD were 53 +/- 20 ms in idiopathic VT patients, 92 +/- 20 ms in organic VT patients, 46 +/- 18 ms in normal individuals, respectively. A positive TWA test result was seen more (P <.01) frequently, and QTD was longer (P <.01) in organic VT patients compared to normal individuals, whereas there was no difference between idiopathic VT patients and normal individuals. In addition, in patients with idiopathic VT, neither did any of these measurements differ between patients with sustained VT (lasting for > or =30 s) and those with nonsustained VT. Noninvasive markers of repolarization inhomogeneity, such as TWA and QTD, are not useful for identifying patients with idiopathic VT. Repolarization inhomogeneity may not affect to the pathogenesis of idiopathic VT.

[Peripheral vascular pain].

Peripheral vascular pain consists of complex factors, and may be divided into three types, i.e., arterial pain, microvessel pain and venous pain. Among these, arterial pain requires intensive pain control because of severe ischemic pain due to arterial obstruction. Under ischemic condition, adenosine is generated, and activates unmyelinated afferents to produce pain. In addition to adenosine, acidic pH itself produces pain and sensitization to mechanical stimuli. Moreover bradykinin generated by kallikurein in acidic pH can produce pain. Nerve block is indicated to improve tissue circulation and to relieve pain. Endoscopic thoracic sympathectomy is indicated for upper extremities, and high frequency thermocoagulation is applied for lumbar sympathectomy. Spinal cord stimulation and the gene therapy with vascular endothelial growth factor have also been reported effective.

Influence of polyethylene and femoral head surface quality on wear: a retrieval study.

Thirty-two polyethylene sockets and 22 femoral heads were retrieved because of aseptic loosening more than 9 years after total hip arthroplasty. The volumetric wear rates of the retrieved polyethylene sockets were significantly greater in those coupled with an alumina head (P < 0.05). The retrieved alumina heads showed significantly better surface roughness and roundness than heads of Co-Cr and of stainless steel (P < 0.05). However, no significant difference was found in polyethylene quality demonstrated as fusion defects among the three different groups. The present study suggests that maintaining better surface roughness and roundness of the femoral heads does not always result in an in vivo reduction of polyethylene wear.

Promoting effect of colostrum on the phagocytic activity of bovine polymorphonuclear leukocytes in vitro.

Bovine colostrum contains a variety of essential nutrients, antibodies, cytokines, hormones, and growth factors that are important for nutrient supply, host defense, growth and for general neonatal adaptation. We have investigated the effect of bovine colostrum on the phagocytic activity for latex particles by normal peripheral blood polymorphonuclear leukocytes using flow cytometric analysis. The phagocytosis promoting effect was observed in colostrum. The promoting activity showed dose-dependent increase up to 25% at a concentration of colostrum. This activity was maximum in the colostrum obtained at parturition and gradually decreased with the time course of lactation as well as IgG level. Colostrum possessed the higher promoting activity than milk and normal serum. These results indicate that bovine colostrum strongly activates polymorphonuclear leukocyte phagocytosis, suggesting the concernment with development of nonspecific immune system in newborns.

Effect of segmental transcatheter arterial chemoembolization on branched chain amino acids and tyrosine ratio in patients with hepatocellular carcinoma.

The effect of segmental transcatheter arterial chemoembolization (TAE) on serum amino acid levels and liver function were studied in 23 patients with HCC associated with hepatitis virus C (22 patients) or alcoholism (1 patient), with compensated liver cirrhosis (Child A 18 patients, Child B 5 patients). Serum levels of branched-chain amino acids (BCAA), tyrosine, branched-chain amino acids to tyrosine ratio (BTR), ammonia, total bilirubin and albumin, and prothrombin times were measured before and after TAE (24 h, 7 and 14 days). The BTR was increased significantly 24 h after TAE (p<0.001) and gradually decreased to pre-TAE levels. Serum tyrosine levels decreased at 24 h after TAE (p<0.005) and later increased. Serum BCAA levels increased slightly at 7days after TAE and were decreased at 14 days after TAE. This results indicated that the increased BTR was due primarily to the decreased tyrosine level at 24 h after TAE. Serum ammonia levels gradually decreased after TAE and the prothrombin time and serum levels of total bilirubin and albumin were not significantly changed. In this study, segmental TAE had little influence on liver function, and the BTR unexpectedly increased at 24 h after TAE. These results suggest that segmental TAE has minimal side effects and may have a beneficial effect on amino acid metabolism.

Atherosclerosis and omega-3 fatty acids in the populations of a fishing village and a farming village in Japan.

The effect of different dietary habits on atherosclerosis was investigated by examining the content of ordinary diets and relevant risk factors through a mass health survey on two village populations in Japan. In total, 261 inhabitants in the fishing village and 209 in the farming village were examined for body build, blood pressure, and blood chemistry. Information on smoking habits and food consumption was obtained using a semi-quantitative item-frequency questionnaire. Pulse wave velocity of the aorta, intima-media thickness of the carotid artery, and atherosclerotic plaques as obtained by ultrasonography were used as measures of atherosclerosis. All measures of atherosclerosis are lower in the fishing village than in the farming village in both men and women. There is a striking 5-8-fold difference in the number of atherosclerotic plaques (P < 0.0001) between the populations. The observed differences in atherosclerosis parallels differences in dietary habits and differences in the serum essential fatty acids. Evaluation of the omega-3 fatty acids over the combined populations reveals a negative association with the number of plaques in the common carotid while the omega-6 fatty acids shows a weak positive association with plaques.

Effects of tranilast on cultured rabbit corneal keratocytes and corneal haze after photorefractive keratectomy.

PURPOSE: In vitro and in vivo studies were performed to elucidate the effects of tranilast on cellular proliferation and collagen synthesis. METHODS: Subculturing was carried out using keratocytes from rabbits that underwent photorefractive keratectomy (PRK) and developed corneal haze, and keratocytes from normal rabbit cornea. RESULTS: Tranilast suppressed proliferation in cultured keratocytes from the corneal haze region at doses of 30 and 300 micromol/L and collagen synthesis at doses of 3, 30, and 300 micromol/L. Normal corneal cultures showed suppression of keratocyte proliferation and collagen synthesis only at a high dose of tranilast (300 micromol/L). Betamethasone suppressed proliferation of keratocytes in both haze and normal cornea at a dose of 10 micromol/L, as well as collagen synthesis at respective doses of 1 and 10 micromol/L. Diclofenac sodium suppressed collagen synthesis of keratocytes in haze cornea at a high dose of 100 micromol/L, and in keratocytes in normal cornea, at doses of 10 and 100 micromol/L. In an in vivo study, either 0.5% tranilast, 0.1% betamethasone phosphate eye drops, or a tranilast base solution (control) was instilled four times daily to rabbits that had undergone PRK. Weekly evaluation of the inhibitory effect of these drugs on the development of haze was performed 2 weeks after surgery. Tranilast suppressed haze 6-13 weeks after PRK, but betamethasone phosphate showed no effect. CONCLUSION: These results indicate that tranilast is potentially effective for inhibiting the corneal haze that occurs after PRK.

The effect of ketamine on clinical endpoints of hypnosis and EEG variables during propofol infusion.

BACKGROUND: We studied the effect of variable doses of ketamine on the endpoints of hypnosis, e.g., unresponsiveness to verbal commands (UVC), loss of eyelash reflex (LER), and inhibition of body movement response with or without sneezing to nasal membrane stimulation (INBMR), and processed EEG variables, e.g., bispectral index (BIS), 95% spectral edge frequency (SEF) and median frequency (MF) during propofol infusion. METHODS: Forty-eight patients received either propofol infusion, 30 mg.kg-1.h-1 (Group P; n = 12) or ketamine bolus, 0.25, 0.5 or 0.75 mg i.v., followed by propofol infusion, 30 mg.kg-1.h-1 + variable dose ketamine infusion, 0.25, 0.5 or 0.75 mg.kg-1.h-1 (Groups PK0.25, PK0.5 and PK0.75; n = 12 each) until UVC, LER and INBMR. BIS, 95% SEF and MF values were monitored and recorded at the endpoints of hypnosis. Propofol and ketamine concentrations were measured at INBMR. RESULTS: Propofol infusion, 30 mg.kg-1.h-1, induced UVC, LER and INBMR at BIS: 65 +/- 2, 63 +/- 9 and 33 +/- 7; 95% SEF: 17 +/- 3, 17 +/- 4 and 14 +/- 3; and MF values of 5 +/- 2, 5 +/- 3 and 3 +/- 2, respectively. With adjunctive ketamine (Groups PK0.5 and PK0.75), the hypnotic endpoints were achieved at higher BIS and 95% SEF values and lower propofol doses and concentrations as compared to Groups P and PK0.25 (9.9 +/- 5.8 and 9.4 +/- 3.4 vs. 13.4 +/- 4.5 and 14 +/- 5.8 micrograms.ml-1). CONCLUSIONS: Our results suggest additive interaction between propofol and ketamine (Groups PK0.5 and PK0.75) for achieving the hypnotic endpoints; however, ketamine did not depress the EEG variables in proportion to its hypnotic effect. The paradoxically higher BIS and 95% SEF values at the hypnotic endpoints may be due to lower propofol concentrations and/or no effect of ketamine on the EEG variables.

Antithrombotic activity of avian crown dermatan sulfate.

The antithrombotic activity of dermatan sulfate from avian crown with the mean molecular weight of 38000 was compared to those from bovine intestine with the mean molecular weight of 16000 in vivo and in vitro. In an in vitro test, bovine intestine dermatan sulfate exhibited stronger effects on stimulation of heparin cofactor II and activation of Glu-plasminogen by tissue plasminogen activator. In vivo, avian crown dermatan sulfate more effectively prevented the development of thrombus in a rat deep vein thrombosis model. The measurement of plasma levels of these two kinds of dermatan sulfate revealed that avian crown dermatan sulfate circulated in higher concentration and longer duration than bovine intestine dermatan sulfate after intravenous administration to rats.

Relationships between 4-hydroxy-2-nonenal, 2-thiobarbituric acid reactive substances and n-6 polyunsaturated fatty acids in refrigerated and frozen pork.

The relationships between 4-hydroxy-2-nonenal (HNE), 2-thiobarbituric acid reactive substances (TBARS) and n-6 polyunsaturated fatty acids (n-6 PUFAs) were investigated for pork stored at 0, -20 and -80 degrees C. A significant linear correlation was apparent between HNE and TBARS for the pork stored at each temperature. However, the n-6 PUFA content remained unchanged during storage.

[Meige's syndrome associated with basal ganglia and thalamic functional disorders].

Magnetic resonance imaging (MRI) or single positron emission computed tomography (SPECT) or both were performed and the responses of surface electromyography (EMG) were examined in seven cases of Meige's syndrome. MRI or SPECT or both demonstrated lesions of the basal ganglia, the thalamus, or both in five of the cases. Surface EMG revealed abnormal burst discharges in the orbicularis oculi and a failure of reciprocal muscular activity between the frontalis and orbicularis oculi in all the cases. These findings suggest that voluntary motor control and reciprocal activity in the basal ganglia-thalamocortical circuits are impaired in Meige's syndrome. In addition, good responses were seen to clonazepam, tiapride and trihexyphenidyl in these cases. Therefore, we conclude that dopaminergic, cholinergic, and gamma-aminobutyric acid (GABA) ergic imbalances in the disorders of the basal ganglia and thalamus in Meige's syndrome cause control in the excitatory and inhibitory pathways to be lost, resulting in the failure of integration in reciprocal muscular activity and voluntary motor control. This failure subsequently causes the symptoms of Meige's syndrome.