Effects of hydrogen-rich water and ascorbic acid treatment on spontaneously hypertensive rats.

Hydrogen-rich water (HW) has been suggested to possess antioxidant properties of value in treatments of lifestyle diseases and for prevention of latent pathologies. To date, the potential benefits of HW against the deleterious effects of excessive salt intake and hypertension have not been investigated. Here, we first examined the effects of HW or HW supplemented with 0.1% ascorbic acid (HWA) on spontaneously hypertensive rats (SHR) that had been fed a normal diet. In comparison to control rats given distilled water (DW), we found that HW did not significantly influence systolic blood pressure (SBP) or diastolic blood pressure (DBP) in SHR; however, the increase in SBP and DBP were inhibited in the HWA group. Next, four groups of SHR were given DW, 0.1% ascorbic acid-added DW (DWA), HW, or HWA in combination with a 4% NaCl-added diet. SHR fed the 4% NaCl-added diet showed increased hypertension; HWA treatment resulted in a significant reduction in blood pressure. The HWA group tended to have lower plasma angiotensin II levels than the DW group. In addition, urinary volumes and urinary sodium levels were significantly lower in the HWA group than the DW group. Urinary isoprostane, an oxidative stress marker, was also significantly lower in the HWA group, suggesting that the inhibitory effect of HWA on blood pressure elevation was caused by a reduction in oxidative stress. These findings suggest a synergistic interaction between HW and ascorbic acid, and also suggest that HWA ingestion has potential for prevention of hypertension.

Effects of cyclic tensile strain and microgravity on the distribution of actin fiber and Fat1 cadherin in murine articular chondrocytes.

Chondrocytes as mechano-sensitive cells can sense and respond to mechanical stress throughout life. In chondrocytes, changes of structure and morphology in the cytoskeleton have been potentially involved in various mechano-transductions such as stretch-activated ion channels, integrins, and intracellular organelles. However, the mechanism of cytoskeleton rearrangement in response to mechanical loading and unloading remains unclear. In this study, we exposed chondrocytes to a physiological range of cyclic tensile strain as mechanical loading or to simulated microgravity by 3D-clinostat that produces an unloading environment. Based on microarray profiling, we focused on Fat1 that implicated in the formation and rearrangement of actin fibers. Next, we examined the relationship between the distribution of Fat1 proteins and actin fibers after cyclic tensile strain and microgravity. As a result, Fat1 proteins did not colocalize with actin stress fibers after cyclic tensile strain, but accumulated near the cell membrane and colocalized with cortical actin fibers after microgravity. Our findings indicate that Fat1 may mediate the rearrangement of cortical actin fibers induced by mechanical unloading.

Fatty Acid Composition and Applications of Eriobotrya japonica Seed Oil.

The loquat (Eriobotrya japonica) is commonly found in Japan. Its fruits are consumed raw or used in processed foods, and its leaves are used as a traditional medicine and in the manufacturing of cosmetics. Additionally, its seeds have several industrial applications. Therefore, this study aimed to estimate the fatty acid composition of loquat seed oil, and to evaluate its potential application as a deodorant. Palmitic acid, linoleic acid, behenic acid, and lignoceric acid were found to be the primary fatty acids present in the seeds, among which linoleic acid was involved in the deodorization of allyl methyl sulfide. Based on these results, loquat seed oil has potential for use in deodorant production.

Transcutaneous Carbon Dioxide Improves Contractures After Spinal Cord Injury in Rats.

BACKGROUND: Joint contractures are a major complication in patients with spinal cord injuries. Positioning, stretching, and physical therapy are advocated to prevent and treat contractures; however, many patients still develop them. Joint motion (exercise) is crucial to correct contractures. Transcutaneous carbon dioxide (CO2) therapy was developed recently, and its effect is similar to that of exercise. This therapy may be an alternative or complementary approach to exercise. QUESTION/PURPOSES: Using an established model of spinal cord injury in rats with knee flexion contractures, we sought to clarify whether transcutaneous CO2 altered (1) contracture, as measured by ROM; (2) muscular and articular factors contributing to the loss of ROM; (3) fibrosis and fibrosis-related gene expression in muscle; and (4) the morphology of and fibrosis-related protein expression in the joint capsule. METHODS: Thirty-six Wistar rats were divided into three equal groups: caged control, those untreated after spinal cord injury, and those treated with CO2 after spinal cord injury. The rats were treated with CO2 from either the first day (prevention) or 15th day (treatment) after spinal cord injury for 2 or 4 weeks. The hindlimbs of rats in the treated group were exposed to CO2 gas for 20 minutes once daily. Knee extension ROM was measured with a goniometer and was measured again after myotomy. We calculated the muscular and articular factors responsible for contractures by subtracting the post-myotomy ROM from that before myotomy. We also quantified histologic muscle fibrosis and evaluated fibrosis-related genes (collagen Type 1, α1 and transforming growth factor beta) in the biceps femoris muscle with real-time polymerase chain reaction. The synovial intima's length was measured, and the distribution of fibrosis-related proteins (Type I collagen and transforming growth factor beta) in the joint capsule was observed with immunohistochemistry. Knee flexion contractures developed in rats after spinal cord injuries at all timepoints. RESULTS: CO2 therapy improved limited-extension ROM in the prevention group at 2 weeks (22° ± 2°) and 4 weeks (29° ± 1°) and in the treatment group at 2 weeks (31° ± 1°) compared with untreated rats after spinal cord injuries (35° ± 2°, mean difference, 13°; 39° ± 1°, mean difference, 9°; and 38° ± 1°, mean difference, 7°, respectively) (95% CI, 10.50-14.86, 8.10-10.19, and 4.73-9.01, respectively; all p < 0.001). Muscular factors decreased in treated rats in the prevention group at 2 weeks (8° ± 2°) and 4 weeks (14°± 1°) and in the treatment group at 2 weeks (14 ± 1°) compared with untreated rats (15° ± 1°, 4.85-9.42; 16° ± 1°, 1.24-3.86; and 17° ± 2°, 1.16-5.34, respectively; all p < 0.05). The therapy improved articular factors in the prevention group at 2 weeks (4° ± 1°) and 4 weeks (6° ± 1°) and in the treatment group at 2 weeks (8° ± 1°) compared with untreated rats (10° ± 1°, 4.05-7.05; 12° ± 1°, 5.18-8.02; and 11° ± 2°, 1.73-5.50, respectively; all p < 0.05). CO2 therapy decreased muscle fibrosis in the prevention group at 2 weeks (p < 0.001). The expression of collagen Type 1, α1 mRNA in the biceps femoris decreased in treated rats in the prevention group at 2 and 4 weeks compared with untreated rat (p = 0.002 and p = 0.008, respectively), although there was little difference in the expression of transforming growth factor beta (p > 0.05). CO2 therapy did not improve shortening of the synovial intima at all timepoints (all p > 0.05). CO2 therapy decreased transforming growth factor beta immunolabeling in joint capsules in the rats in the prevention group at 2 weeks. The staining intensity and Type I collagen pattern showed no differences among all groups at all timepoints. CONCLUSION: CO2 therapy may be useful for preventing and treating contractures after spinal cord injuries. CO2 therapy particularly appears to be more effective as a prevention and treatment strategy in early-stage contractures before irreversible degeneration occurs, as shown in a rat model. CLINICAL RELEVANCE: Our findings support the idea that CO2 therapy may be able to improve the loss of ROM after spinal cord injury.

Antihypertensive Effect of γ-Aminobutyric Acid-Enriched Brown Rice on Spontaneously Hypertensive Rats.

'Haiibuki' is a giant embryo rice cultivar that contains abundant γ-aminobutyric acid (GABA) compared with conventional rice cultivars. Here, we performed a functional evaluation of 'GABA-enriched brown rice' (GEBR) prepared by modifying the 'Haiibuki' cultivar to contain more GABA. Study 1: Spontaneously hypertensive rats were divided into three groups [control (cornstarch), normal brown rice, and GEBR] and fed an orally administered diet for 4 wk. A significant blood pressure elevation-inhibitory effect was observed in the GEBR group as compared with the other groups. Study 2: Rats were divided into two groups and fed ad libitum for 12 wk. The two groups were control (commercial feed with 5% cornstarch) and GEBR (commercial feed with 5% GEBR). Body weight, blood pressure, food consumption, and water intake were measured during the study period, and blood chemistry was analyzed after the study. Plasma 8-hydroxy-2'-deoxyguanosine (8-OHdG) and urinary isoprostane were measured 12 and 10 wk after the start of the study, respectively. A significant blood pressure elevation-inhibitory effect was observed in the GEBR group. The 8-OHdG and isoprostane levels were significantly lower in the GEBR group than in the control group, demonstrating an oxidative stress-reducing effect. Therefore, GEBR exhibited a blood pressure elevation-inhibitory effect under the conditions of this study. The antioxidative action may occur secondarily to the antihypertensive action of GABA, suggesting that the long-term ad libitum ingestion of GEBR prevents hypertension. A reduction in oxidative stress could reduce the chances of complications in cardiovascular diseases.

Mechanical Stress by Spasticity Accelerates Fracture Healing After Spinal Cord Injury.

Accelerated fracture healing in patients with spinal cord injuries (SCI) is often encountered in clinical practice. However, there is no distinct evidence in the accelerated fracture healing, and the mechanisms of accelerated fracture healing in SCI are poorly understood. We aimed to determine whether SCI accelerated fracture healing in morphology and strength, to characterize the healing process with SCI, and to clarify the factors responsible for accelerated fracture healing. In total, 39 male Wistar rats were randomly divided into healthy control without intervention, SCI only, fracture with SCI, botulinum toxin (BTX) A-treated fracture with SCI, and propranolol-treated fracture with SCI groups. These rats were assessed with computed microtomography, histological, histomorphological, immunohistological, and biomechanical analyses. Both computed microtomography and histological analyses revealed the acceleration of a bony union in animals with SCI. The strength of the healed fractures after SCI recovered to the same level as that of intact bones after SCI, while the healed bones were weaker than the intact bones. Immunohistology revealed that SCI fracture healing was characterized by formation of callus with predominant intramembranous ossification and promoting endochondral ossification. The accelerated fracture healing after SCI was attenuated by BTX injection, but did not change by propranolol. We demonstrated that SCI accelerate fracture healing in both morphology and strength. The accelerated fracture healing with SCI may be due to predominant intramembranous ossification and promoting endochondral ossification. In addition, our results also suggest that muscle contraction by spasticity accelerates fracture healing after SCI.

Stretching After Heat But Not After Cold Decreases Contractures After Spinal Cord Injury in Rats.

BACKGROUND: Contractures are a prevalent and potentially severe complication in patients with neurologic disorders. Although heat, cold, and stretching are commonly used for treatment of contractures and/or spasticity (the cause of many contractures), the sequential effects of these modalities remain unclear. QUESTIONS/PURPOSES: Using an established rat model with spinal cord injury with knee flexion contracture, we sought to determine what combination of heat or cold before stretching is the most effective for treatment of contractures derived from spastic paralyses and investigated which treatment leads to the best (1) improvement in the loss of ROM; (2) restoration of deterioration in the muscular and articular factors responsible for contractures; and (3) amelioration of histopathologic features such as muscular fibrosis in biceps femoris and shortening of the joint capsule. METHODS: Forty-two adolescent male Wistar rats were used. After spasticity developed at 2 weeks postinjury, each animal with spinal cord injury underwent the treatment protocol daily for 1 week. Knee extension ROM was measured with a goniometer by two examiners blinded to each other's scores. The muscular and articular factors contributing to contractures were calculated by measuring ROM before and after the myotomies. We quantitatively measured the muscular fibrosis and the synovial intima length, and observed the distribution of collagen of skeletal muscle. The results were confirmed by a blinded observer. RESULTS: The ROM of heat alone (34° ± 1°) and cold alone (34° ± 2°) rats were not different with the numbers available from that of rats with spinal cord injury (35° ± 2°) (p = 0.92 and 0.89, respectively). Stretching after heat (24° ± 1°) was more effective than stretching alone (27° ± 3°) at increasing ROM (p < 0.001). Contrastingly, there was no difference between stretching after cold (25° ± 1°) and stretching alone (p = 0.352). Stretching after heat was the most effective for percentage improvement of muscular (29%) and articular (50%) factors of contractures. Although quantification of muscular fibrosis in the rats with spinal cord injury (11% ± 1%) was higher than that of controls (9% ± 0.4%) (p = 0.01), no difference was found between spinal cord injury and each treatment protocol. The total synovial intima length of rats with spinal cord injury (5.9 ± 0.2 mm) became shorter than those of the controls (7.6 ± 0.2 mm) (p < 0.001), and those of stretching alone (6.9 ± 0.4 mm), stretching after heat (7.1 ± 0.3 mm), and stretching after cold (6.7 ± 0.4 mm) increased compared with rats with spinal cord injury (p = 0.01, p = 0.001, and p = 0.04, respectively). The staining intensity and pattern of collagen showed no difference among the treatment protocols. CONCLUSIONS: This animal study implies that heat or cold alone is ineffective, and that stretching is helpful for the correction of contractures after spinal cord injury. In addition, we provide evidence that heat is more beneficial than cold to increase the effectiveness of stretching. CLINICAL RELEVANCE: Our findings tend to support the idea that stretching after heat can improve the loss of ROM and histopathologic features of joint tissues. However, further studies are warranted to determine if our findings are clinically applicable.

Evaluation of the fatty acid composition of the seeds of Mangifera indica L. and their application.

Mango (Mangifera indica L.), an edible fruit, is one of the main agricultural products in many tropical regions. Mango varieties differ in not only fruit shape but also aroma, which is an important characteristic. Although the fruit has many uses, the seeds are discarded as waste. Therefore, this study aimed to estimate the fatty acid content of seed oil of mangoes from different cultivation areas (Miyazaki, Japan, and Taiwan), and to evaluate their application in cosmetics. Five fatty acids were identified in the mango seed oil. Oleic acid and stearic acid were the principal components of mango seed oil obtained from Miyazaki (46.1% and 39.8%, respectively) and Taiwan (43.7% and 40.1%, respectively). As a cosmetic ingredient, mango seed oil showed good deodorizing effect on both 2-nonenal and isovaleric acid. The results indicated the potential applications of mango seed oil in the cosmetic industry.

Isolation and identification of flavonoids from Coreopsis lanceolata L. petals.

The methanol extract of Coreopsis lanceolata L. petals was acid-hydrolysed, and 7,3',4'-trihydroxy-8-methoxyflavanone (1) and 6,3',4'-trihydroxy-7-methoxyaurone (leptosidin) (2) were successfully isolated. The structure of compound 1 is designated to flavanone based on X-ray crystallographic analysis and NMR spectroscopic analysis. Compound 1 showed high-antioxidant effects based on diphenylpicrylhydrazyl radical scavenging assay (94.3% scavenging rate) and superoxide dismutase-like activity assay (23.9% inhibition rate).

Color preferences of laboratory mice for bedding materials: evaluation using radiotelemetry.

Preferences for different housing conditions in mice were evaluated by radiotelemetry. Male C57BL/6J and ICR mice were used. Preference for bedding materials in mice was compared among three materials, wood shavings (WS), paper (CF) and cloth (AG), using the length of stay in cages as a parameter. The results indicated that mice stayed longer in a cage with AG than in cages with other bedding materials. The present study confirmed our previous results and thereby indicated that radiotelemetry is a useful method to evaluate impacts of housing conditions on animal welfare. In the second part of this study, we used radiotelemetry to evaluate color preference of the mice for cloth bedding material. In C57BL/6J mice, staying time in black cloth was significantly longer than that in white cloth. In ICR mice, staying time in white cloth was significantly longer than that in black cloth. The mice preferred the environment with the same color as their fur, which may be important for animal welfare.

A study on fatty acids in seeds of Euterpe oleracea Mart seeds.

Fatty acids of Euterpe oleracea Mart seeds were analyzed in hexane and diethyl ether extracts. The hexane extract contained dodecanoic acid (3), tetradecanoic acid (4), hexadecanoic acid (7), and 9-octadecenoic acid (10). The diethyl ether extract contained (10). The oil of Mart seeds was obtained firstly by methanol extract and further extracted with diethyl ether. The hexane and diethyl ether extracts were then analyzed for antioxidant effects. Both extracts demonstrated a 1,1-diphenyl-2-picrylhydrazyl radical elimination effect similar to that of α-tocopherol and an active oxygen inhibition effect. Significant quantities (0.6212 mg/mL) of polyphenol, in comparison to the standard rejected gallic acid, were found in the extract oil of Mart seeds by the methanol extraction.

Antioxidant activity of colored rice bran obtained at different milling yields.

In this study, we investigated the antioxidant components of three types of colored rice bran--forbidden rice, red rice and green rice--obtained from rice in which the pigment layer had been removed at milling yields of 90%-100% and 80%-90%. An evaluation of the effects of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity revealed that rice bran obtained from forbidden rice at milling yields of 90%-100% and 80%-90% and rice bran obtained from red rice at milling yields of 90%-100% showed favorable antioxidant activity. The antioxidant components were confirmed to be 3,4-dihydroxy methyl benzoate and p-methoxyphenol and they influence the antioxidant activity of the three types of colored rice bran.

Suppressive ability of defatted rice bran against lipid oxidation in cookies containing iron.

Cookies containing iron, defatted rice bran, and several oils were prepared, and their oxidative stability evaluated. Oxidation was suppressed by the defatted rice bran, but a limit to the suppressive effect was observed. The maximum peroxide values obtained with defatted rice bran were low, and similar, regardless of the degree of unsaturation of the oils. A chemical analysis suggested that proteins and polyphenols in the defatted rice bran contributed to the suppressive effect. Cookies without the defatted rice bran showed decreasing maximum peroxide value as the relative humidity increased. No such dependence was observed for the cookies containing defatted rice bran. The water sorption isotherm of defatted rice bran indicated that the weak dependence was due to low water sorption.

Synthesis of acyl arbutin by an immobilized lipase and its suppressive ability against lipid oxidation in a bulk system and O/W emulsion.

Acyl arbutin was synthesized through the condensation of arbutin with a saturated fatty acid (C6-18) by the immobilized lipase in a batch reaction. The conversion at 10 and 20 g/l-solvent of immobilized lipase reached 45% over 2 d, but the initial reaction rate per amount of immobilized lipase decreased at 20 g/l-solvent. The radical scavenging activity of acyl arbutin in an ethanol solution was independent of the acyl chain length, although the rate constant, k, estimated for the oxidation of methyl linoleate in a bulk system with acyl arbutin by using the Weibull equation, decreased as the acyl chain length increased. This indicates the antioxidative ability of acyl arbutin with a long acyl chain to be due to its lipophilicity. Furthermore, it is suggested that dodecanoyl arbutin mainly acted on the interface between the oil and water phases in an O/W emulsion, and effectively suppressed the oxidation induced at the interface.

Pharmacological activity of compounds extracted from persimmon peel (Diospyros kaki THUNB.).

Persimmon peels (Diospyros kaki THUNB.) are discarded during the production of dried fruit. The 2-methoxy-4-vinylphenol (8) which is component of persimmon peel had high antioxidant activity on the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging assay and SOD (superoxide dismutase) assay. And (8) had higher tyrosinase inhibiting activity than that of arbutin using both L-tyrosine and L-DOPA as substrates. In addition, tyrosinase inhibiting activity of synthesized 2-methoxy-4-vinylphenol glycoside (8a) was studied. (8a) had tyrosinase inhibiting activity, suggesting that (8a) has possibilities for ingredient of cosmetics that are possessed of whitening effect.

Chemical constituents of Coreopsis lanceolata L. and their physiological activities.

Petals from Coreopsis lanceolata L. were extracted in methanol and subjected to solvent-solvent partitioning to yield four fractions: hexane, diethyl ether, ethyl acetate, and aqueous. These fractions were screened using a 1,1-diphenyl-2-picrylhydrazyl radical scavenging assay and an active oxygen inhibition assay. The ethyl acetate extract contained the highest level of active components. Four compounds isolated from this fraction by HPLC were screened for antioxidant activity and anti-allergic activity. Anti-allergic activity was assessed by measuring release of histamine from rat peritoneal mast cells. One compound, termed laceolin, was characterized by spectroscopic analysis, including extensive NMR and mass spectra. This compound had significant radical scavenging activity, with an SC50 (scavenging concentration of 50%) value of 2.6 microg/mL and superoxide dismutase activity of 46.2%.

Suppression of mutagens-induced SOS response by phytoncide solution using Salmonella typhimurium TA1535/pSK1002 umu test.

Four types of phytoncide solution (A-Type, AB-Type, D-Type and G-Type) were evaluated as antimutagenic agents with suppressive effects on the SOS-inducing activity of the mutagen 2-(2-furyl)-3-(5-nitro-2-furyl) acrylamide (furylfuramide) using Salmonella typhimurium TA1535/pSK1002 umu test. The A-Type, AB-Type, D-Type and G-Type of phytoncide solution suppressed the SOS-inducing activity on furylfuramide at a concentration of 100 microg/mL by 86.1%, 74.7%, 69.5% and 55.4%, respectively, and the ID(50) (50% inhibitory dose) values were 9.0 microg/mL, 22.5 microg/mL, 36.0 microg/mL and 72.8 microg/mL. They also showed the suppression of SOS-inducing activity against other chemical mutagens, such as 4-nitroquinolin 1-oxide (4NQO) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), which do not require liver metabolizing enzymes, and against 2-aminoanthracene (2AA) and 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), which require these enzymes, and against UV irradiation, which is a well known physical mutagen. In the search for the component-activity relationship, the A-Type of phutoncide solution suppressed the SOS-inducing activity greater than the other types of phutoncide solution for furylfuramide, 4NQO and MNNG. However, in case of 2AA and Trp-P-1, the D-Type of phytoncide solution was most effective in suppressing the SOS-inducing activity in the umu test. From these results, the four types of phytoncide solutions showed the suppressive effect of SOS-inducing activity against chemical and physical mutagens.

Predictive factors for anastomotic leakage in the neck after retrosternal reconstruction for esophageal cancer.

BACKGROUND/AIMS: Anastomotic leakage after transthoracic esophagectomy for esophageal cancer can induce life-threatening morbidity. This study investigated the predictive factors for anastomotic leakage in the neck after retrosternal reconstruction. METHODOLOGY: A total of 129 esophageal carcinoma patients undergoing transthoracic esophagectomy and esophagogastric anastomosis in the neck via a retrosternal approach were enrolled between April 1985 and March 2002. Predictive factors for anastomotic leakage were statistically evaluated. In a preliminary study using 18 cases, thoracic inlet space (TIS) was recommended to be extended more than 700 mm2. RESULTS: Partial resection of the bony structures was performed in 32 patients. The method of anastomosis and partial resection of bony structures according to the TIS independently influenced the likelihood of anastomotic leakage, with hand-sewn anastomosis and an absence of partial resection increasing its occurrence. CONCLUSIONS: Stapled anastomosis following the partial resection of the sternum and the left clavicle is recommended to avoid anastomotic leakage. These findings should be clarified by a randomized controlled study in a high-volume hospital.

Antioxidant effects and antimicrobial activites of phytoncide.

Physiological activities of four types of phytoncide solutions (A, AB, CY and D-types), prepared from various plants widely distributed in nature, were examined. We assayed these phytoncide solutions, testing for active oxygen inhibition, DPPH radical scavenging effects, nitrogen monoxide inhibition, and lipid peroxide inhibition. The AB- and D-types of phytoncide solutions especially showed comparably potent antioxidant effects. Antibacterial and antifungal assays were conducted using the AB- and D-types, with results showing significant inhibitory activities with these solutions.

Synthesis and physiological activity of thiophenes and furans with 3- and 4-methoxyacetophenone derivatives.

The synthesis and physiological activity of thiophenes and furans with methoxyacetophenone derivatives were examined. 3-Methoxyacetophenone (1) and 4-methoxyacetophenone (2) were converted, respectively, to the oximes (3) and (4) by oximation with hydroxylamine hydrochloride, and to the primary amines (5) and (6) by reduction with LiAlH(4). The primary amine derivatives were further converted into the thiophene and furan compounds (5a) approximately (5h) and (6a) approximately (6h), respectively. Bioassay of these compounds (5a) approximately (5h) and (6a) approximately (6h) on the germination of lettuce(Lactuca satiba) seeds showed that compound (5b) exhibits growthpromoting activity.