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Quantitative analysis of phenolic compounds in Aloe arborescens fermented extract revealed that barbaloin concentration was found to be 14 μg/mL (14 ppm) by HPLC analysis. Seven phenolic compounds could be identified: aloesin, 8-C-glycosyl-7-O-methyl-S-aloesol, aloenin, isobarbaloin, barbaloin, 2 -O-feruloylaloesin, and aloe-emodin based on previous HPLC analysis. Short chain fatty acids such as acetic, propionic, butyric, lactic acid, were identified in the fermented extract of A. arborescens by GC/MS analysis. Reduction of barbaloin concentration and production of butyric acid clearly indicate a potential efficacious role of endophytic bacteria in A. arborescens leaves. Dietary intake of the extract fermented by endophytic bacteria in A. arborescens highlights immune-stimulating activity to human health and strongly influences an importance of host-microbial crosstalk to ensure maintenance of homeostasis.

Aim: The present study was conducted to compare effects of acemannan and formocresol as pulp-dressing agents clinically and radiographically in primary teeth. Materials and Methods: Aloe vera gel high molecular weight fractions (AHM) containing native ingredients of polysaccharide (acemannan) and glycoprotein (verectin) were obtained by using the patented hyper dry system after washing out coloured materials with running water from Aloe vera gel slurry. Thirty healthy children aged from four to eight years, they were selected from patients attending outpatient clinic of Pediatric Dentistry department. Each child had at least bilateral deep carious lower primary molar indicated for pulpotomy. Pulpotomy was done in both groups; where group I treated by AHM, acemannan, as a dressing agent while formocresol used as a dressing agent in group II. The study cases were recalled after three, six, nine and twelve months for clinical and radiographic evaluation. Results: The overall clinical success rate of acemannan group was 96.5%, while formocresol group was 89.6%. The two groups were clinically successful with no statistically significant difference between them (p = 0.148). The radiographic success rate of acemannan group was 93.1%, while for formocresol group was 86.2%. There was no statistically significant difference between the two groups (p = 0.385). Conclusion: The acemannan showed higher clinical and radiographic success rate compared to formocresol as a pulpulpotomy agent in molars. It can be considered as an acceptable biomaterial for vital pulp therapy of deep caries in primary teeth.

Propolis, a resinous substance produced by bees, is used as a folk medicine for treatment of periodontal diseases. However, its mode of the action and the compounds responsible for its activities remain obscure. In the present study, we comprehensively investigated the antibacterial activities of ethanol-extracted propolis (EEP) and EEP-derived compounds toward Porphyromonas gingivalis, a keystone pathogen for periodontal diseases. Broth microdilution and agar dilution assays were used to determine the minimum inhibitory concentrations of EEP against a range of oral bacterial species, of which P. gingivalis showed a higher level of sensitivity than oral commensals such as streptococci. Its antibacterial activity toward P. gingivalis was maintained even after extensive heat treatment, demonstrating a high level of thermostability. EEP also induced death of P. gingivalis cells by increasing membrane permeability within 30 min. Spatiotemporal analysis based on high-speed atomic force microscopy revealed that EEP immediately triggered development of aberrant membrane blebs, followed by bleb fusion events on the bacterial surface. Furthermore, we isolated artepillin C, baccharin, and ursolic acid from EEP as antibacterial compounds against P. gingivalis. Of those, artepillin C and baccharin showed bacteriostatic activities with membrane blebbing, while ursolic acid showed bactericidal activity with membrane rupture. In particular, ursolic acid demonstrated a greater ability to affect bacterial membrane potential with increased membrane permeability, probably because of its highly lipophilic nature as compared with other compounds. Taken together, these findings provide mechanistic insight into the antibacterial activities of EEP and its exquisite membrane-targeting antibacterial compounds and imply the applicability of narrow-spectrum therapeutics with EEP for treatment of periodontitis. In addition, the advanced technology utilized in the present study to visualize the nanometer-scale dynamics of microorganisms will contribute to expanding our understanding of the activities of antimicrobials and the mechanism of drug resistance in bacteria.

The role of gut microbiota and its related activities, like fermentation in immune modulation, are well documented. It was found that acemannan (AM), the extract of aloe leaf gel can exerts significant immune modulating activity. AM has been shown to activate macrophages and stimulate T cells that are involved in the defense against harmful microorganisms by likely acting as an adjuvant for health and disease. However, it should be noted that commercially available AM preparation used for these studies often contains a complex mixture because partially purified carbohydrate preparation containing about 60% acetylated mannan together with other carbohydrates, especially pectin, hemicellulose, and lectins. The beneficial effects of endogenous butyrate productions, delivery, and absorption by colonocytes have been well documented. Mechanistically, butyrate exerts its functions by acting as a histone deacetylase inhibitor or signaling through several G-protein-coupled receptors. Recently, butyrate has received particular attentions for its beneficial effects on intestinal homeostasis and energy metabolism.

Nutraceutical Aloe vera gel scaffolds, such as acemannan, promote soft tissue organization in biomedical applications and polymeric matrix. Aloe vera hydrogels can induce a much-controlled release of active ingredients and be used as bio-functional scaffolds in a biodegradable form. The before or after questionnaire response for locomotive syndrome was administered: Nutraceutical supplement composed of enzyme-hydrolyzed natural eggshell membrane and curcumin, with Aloe vera juice (AVJ) scaffolds, showed the possible improvement of joint comfort and flexibility. The efficacy for pain and stiffness associated with arthralgia of the knee and hip was evaluated based on locomotive syndrome risk test. Furthermore, w-3polyunsaturated fatty acids (EPA and DHA) and L-Arginine with AVJ revealed a possible dietary efficacy for Parkinson’s disease (PD) patients at stage 3. Prophylactic possible efficacies of AVJ as bio-functional scaffolds are fully expected for increasing collagen synthesis in knee and/or hip joint healing and for mitigation of risk factors of PD patients with diabetes.

The involvement of aloe plants in the remedy of chronic diseases has been well publicized in the past. In this review paper, we attempted to focus on the major aloe ingredients on their known efficacy related to the immune modulating activities. Several bioactive phytochemicals present in Aloe barbadensis ( vera) are known to relieve rheumatoid arthritis (RA)-related pains by suppressing chronically over-activated inflammatory process. Aloe-emodin has been shown to promote natural killer cell and macrophage phagocytic activities in the tumor, and emodin, an anthraquinone found also in other plants, appeared to have some protective anti-inflammatory effect by showing its inhibition of IL-1β and LPS-stimulated proliferation of RA synoviocytes in a dose-dependent manner under hypoxic condition. Another putative immune modulator we reviewed in this paper is aloe’s butyrate fermented by the dietary aloe in the gut. Its beneficial effects are based on the host immune metabolism via the activation of intestinal mucosal functions on the suppression of obesity and inflammation. It is hoped that elaboration on butyrate may provide fresh insights as to aloe’s immune modulation and chronic inflammation. Potential efficacious role of barbaloin, aloe-emodin, emodin and fermented butyrate of Aloe vera needs further scrutiny and evidence-based documentation through carefully designed research.

AIM: Long-term dietary intake of non-caloric fiber, such as Aloe vera gel influences the structural and activity of micro-organisms in the human gut. The present investigation was executed with an innovative concept: symbiotic effect of probiotics that are used commercially as lactic acid bacteria i.e. Lactobacillus fermentum and Aloe vera juice as prebiotics for a prospective prophylaxis. MATERIALS AND METHODS: Fermentation of L. fermentum with Aloe vera juice certified by International Aloe Science Council was carried out and the quantification of short chain fatty acids (SCFAs) from the fermentation broth in large scale was determined by gas-chromatography-mass-spectrometry selective detection in the selective ion monitoring mode. RESULTS: The growth of L. fermentum and L. helveticus with Aloe vera juice (AVJ) individually on MRS broth was continued to keep at pH 3.5 and 3.6, while pH of the negative controls changed to 4.3 and 4.0, respectively, during 24 hr incubation. The growth rate and the viability of L. helveticus incubated with different concentrations (5-25%) of Aloe vera juice were strongly reduced. However, the growth rate of L. fermentum was enhanced in a concentration dependent manner with emphasis on the use of 15% AVJ that resulted in two times more growth than that of the negative control. Continuation of L. fermentum growth at pH 3.6 in combination with AVJ during incubation for 24 hr suggests the durability of prebiotic potential by AVJ in in vitro fermentation. Acetic, propionic and lactic acid as SCFAs in the ether extract were identified from the fermentation culture medium. CONCLUSION: The prebiotic activity of AVJ may be assessed by the participation of SCFAs during 24 hrs-incubation with L. fermentum. An innovative concept of symbiotics: a combination of AVJ and L. fermentum, is a perspective prophylaxis on future intestinal health claims. Due to tolerance to acid, L. fermentum may pass through the gastric barrier and colonize the intestine after oral administration.

The health-promoting potential of a balanced gut microbiota status modulated by calorie restriction (CR) posits a possible close link between gut microbiota (GM) and healthy aging. Short chain fatty acids (SCFAs) generated from intestinal bacterial fermentation may act as mediators between the microbiota and the immune system. In this review, a putative prophylactic role of butyric acid from endophytic bacteria fermentation of Aloe vera gel was described to suggest that the consumption of fermented extract of Aloe vera gel may be beneficial for health and QOL as an immune modulator.

AIM: The present study was conducted to compare the histopathological effects of acemannan to that of formocresol as pulp-dressing agents in primary teeth underwent pulpotomy. MATERIALS AND METHODS: Aloe vera gel high molecular weight fractions (AHM) containing native ingredients of polysaccharide (acemannan) and glycoprotein (verectin) were obtained by using the patented hyper dry system after washing out coloured materials with running water from Aloe vera gel slurry. Forty-six primary first teeth from children between 6-9 years, which formerly planned for serial extraction were selected. The children were distributed into two equal groups; twelve teeth for each. Pulpotomy was done in both groups; where group I treated by AHM, acemannan, as a dressing agent while formocresol used as a dressing agent in group II. The teeth from each group were extracted at two, four, and twelve week intervals for histopathological evaluation. RESULTS: In group I (Acemannan), after twelve weeks significantly better results in pulp inflammation, incomplete dentine bridge formation, and soft tissue organization were demonstrated (p

AIM: Diet is a natural source of butyrate through the fermentation of non-digestive fiber, such as acemannan in Aloe vera gel, is a strong appearing target for health and quality of life as an immune modulation and colorectal cancer prevention in aged people. In our earlier research on fermentation by endophytic bacteria in Aloe vera gel, butyric acid was identified by GC/MSD analysis. Present investigation aims the identification of the microbiota. MATERIALS AND METHODS: The endophytic microbiota of Aloe vera gel in the fermented media were examined by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry. RESULTS: The following microbiota were identified: Bacillus cereus, B. licheniformis, Lactobacillus paralimentarium, Yeast: Clavispora lusitaniae. The safety pattern of the prepared Aloe vera gel was tested on normal non-cancerous cells and indicated the absence of any significant possible toxicity on the cells. Also, the extracted gels showed abilities to regulate the inflammatory responses in the inflammation cell models via the reduction in the amount of induced reactive oxygen species and both COX 1 and 2 enzymes. DISCUSSIOIN: Identification of butyrate-producing endophytic microbiota in Aloe vera gel fermentation and finding of inflamatory as well as antioxidant activities of butyrate in the fermented gel may help explain the known beneficial effects of butyrate in intestinal colon and on colitis. An innovative concept of symbiotics: a combination of Aloe vera gel juice and microbiota: Bacillus cereus, B.licheniformis. Lactobacillus paralimentarium and Clavispora lusitaniae, is a perspective on alleviation of cancer disease and improvement of gastrointestinal health by butyrate fermentation.

There is a growing interest in butyrate because its impact on epigenetic mechanisms will lead to more specific and efficacious therapeutic strategies for the presentation and treatment of different disease ranging from genetic/metabolic conditions to neurological degeneration disorders. The dietary natural source of butyrate through a high fiber diet or butyrate produced by fermentation of non-digestive fiber, such as acemannan in Aloe Vera leaf gel, is a highly appealing approach to present a simple and relatively low risk method to potentially improve outcomes in aged people with brain troubles. In this review, we will discuss the pharmacological effects of butyrate as a histone deacetylase inhibitor to an insulin resistance and energy expenditure, and as pro-drugs to ulcerative colitis and cancer, and the gut-liver axis in pre-clinical treatment.

Aloe vera extracts have antimicrobial and anti-fungal activities, which may be able to treat minor skin infections. In spite of these antimicrobial activities, the inner leaf gel containing acemannan: un-digestible polysaccharide, leads to fermentation with endophytic bacteria and results in bacterial growth promotion, while no short chain fatty acids were detected in the ether extract of the fermentation broth with the rind. Butyric acid was identified by GC/MSD analysis from ether extract of the gel fermentation broth, expecting multiple beneficial effects of butyric acid at intestinal and extra-intestinal level. Present investigation exhibits that daily intake of the butyric acid fermentation extract from Aloe vera inner gel with endophytic bacteria, may provide the possible potential preventive and therapeutic roles in human health.

The use of cryosectioning facilitates the morphological analysis and immunocytochemistry of cells in tissues in atomic force microscopy (AFM). The cantilever can access all parts of a tissue sample in cryosections after the embedding medium (sucrose) has been replaced with phosphate-buffered saline (PBS), and this approach has enabled the production of a type of high-resolution image. The images resembled those obtained from freeze-etching replica electron microscopy (EM) rather than from thin-section EM. The AFM images showed disks stacked and enveloped by the cell membrane in rod photoreceptor outer segments (ROS) at EM resolution. In addition, ciliary necklaces on the surface of connecting cilium, three-dimensional architecture of synaptic ribbons, and the surface of the post-synaptic membrane facing the active site were revealed, which were not apparent using thin-section EM. AFM could depict the molecular binding of anti-opsin antibodies conjugated to a secondary fluorescent antibody bound to the disk membrane. The specific localization of the anti-opsin binding sites was verified through correlation with immunofluorescence signals in AFM combined with confocal fluorescence microscope. To prove reproducibility in other tissues besides retina, cryosectioning-AFM was also applied to elucidate molecular organization of sarcomere in a rabbit psoas muscle.

Photochromic diarylethene derivatives bearing hydrophilic N,N-bis(2-(2-(2-methoxyethoxy)ethoxy)ethyl)sulfonamide (SO2NTEG2) (1–3) or myo-inositol (4–6) substituents were synthesized. Although the solubility in aqueous solution of the derivatives containing the four tri(ethylene glycol) chains was relatively poor, the solubility was improved by introducing two myo-inositol residues. The derivatives could be dissolved even in pure water and underwent photochromism upon alternate irradiation with ultraviolet (UV) and visible light, in both organic and aqueous solutions. The absorption maxima of the open- and closed-ring isomers of these derivatives and the photostationary conversion ratios from the open- to the closed-ring isomers upon UV irradiation did not change much, even when the solvent polarity was changed. These results indicate that the diarylethene derivatives are mono-molecularly dissolved in aqueous solution and that intermolecular interactions between the derivatives are negligible when the concentration is less than 1×10−4 mol⋅L−1.

Arundic acid, (R)-(-)-2-propyloctanonic acid, is a novel neurological agent for intractable neurodegenerative diseases. However, arundic acid, an oily drug, has low aqueous solubility and severe bitter/irritating tastes. Consequently, these physicochemical properties of arundic acid need to be improved to develop its pharmaceutical preparations. In the present study, we evaluated whether parent cyclodextrins (CyDs) and 2-hydroxypropylated CyDs (HP-CyDs) can interact with arundic acid, and have powderization, solubilization and taste-masking properties. Of various CyDs, HP-β-CyD had the most potent solubilizing effect for arundic acid. UV and (1)H NMR spectroscopic studies demonstrated that arundic acid formed inclusion complexes with CyDs at a molar ratio of 1:1 in solution. The complexation with CyDs changed an oily form of arundic acid to a solid form. The gustatory sensation studies indicate that of various CyDs, HP-β-CyD and γ-CyD showed the most significant taste-masking effects in solution and powders, respectively. HP-β-CyD significantly reduced the response of the electric potential caused by the adsorption of arundic acid to the taste sensor. These results suggest that hydrophilic CyDs have potential as multifunctional excipients for preparing solutions and powders containing arundic acid.

Aloe vera L. high molecular weight fractions (AHM) containing less than 10ppm of barbaloin and polysaccharide (MW: 1000kDa) with glycoprotein, verectin (MW: 29kDa), were prepared by patented hyper-dry system in combination of freeze–dry technique with microwave and far infrared radiation. AHM produced significant decrease in blood glucose level sustained for 6 weeks of the start of the study. Significant decrease in triglycerides was only observed 4 weeks after treatment and continued thereafter. No deterious effects on kidney and liver functions were apparent. Treatment of diabetic patients with AHM may relief vascular complications probably via activation of immunosystem.

Effects of tanshinone VI, a diterpene from Tan-Shen, on humoral factor-induced phosphorylation of ERK and Akt during hypertrophy of cardiomyocytes and fibrosis of cardiac fibroblasts isolated from neonatal rats were examined. Treatment of cultured cardiomyocytes with 10 nM insulin-like growth factor-1 (IGF-1) or 10 nM endothelin-1 resulted in an increase in leucine incorporation into acid-insoluble fraction. Treatment of cultured cardiac fibroblasts with 10 nM IGF-1 or 10 nM angiotensin II increased incorporation of proline. IGF-1 increased phosphorylated extracellular signal-regulated kinase (pERK) and protein kinase B (pAkt) of cardiomyocytes, whereas endothelin-1 increased pERK, but not pAkt. Treatment of cardiac fibroblasts with 10 nM IGF-1 or 10 nM angiotensin II also increased pERK, whereas pAkt was increased by treatment with IGF-1 alone. When the cardiomyocytes were incubated in the presence of 10 microM tanshinone VI, IGF-1- and endothelin-1-induced increases in pERK, but not pAkt, were partially attenuated. Treatment of cardiac fibroblasts with 10 microM tanshinone VI also attenuated IGF-1-induced increases in pERK and pAkt. Tanshinone VI also partially attenuated angiotensin II-induced increase in proline incorporation into cardiac fibroblasts. PD98059, an inhibitor for phosphorylation of extracellular signal-regulated kinase (ERK), but not wortmannin, that for protein kinase B phosphorylation, attenuated an increase in leucine incorporation into cardiomyocytes in the presence of either IGF-1 or endothelin-1. These results suggest that tanshinone VI is a possible agent that can attenuate the humoral factor-induced hypertrophy of cardiomyocytes and fibrosis of cardiac fibroblasts via an attenuation of ERK phosphorylation in these cells.

Bisphenol A [BPA, 2,2-bis(4-hydoxyphenyl)propane], an industrial chemical used in the production of polycarbonate, epoxide resin, and polyarylate, is considered to be an endocrine-disrupting chemical. BPA may be present in some hollow-fiber dialyzers used in hemodialysis. In this study, we tested the amounts of BPA eluted from various hollow fibers. Furthermore, we measured the BPA concentration in the sera of 22 renal disease predialysis patients, as well as 15 patients who were receiving hemodialysis, to see if there is BPA accumulation in these patients. The elution test of BPA showed that a much larger amount of BPA was eluted from polysulfone (PS), and polyester-polymeralloy hollow fibers. Among renal disease patients who had not undergone hemodialysis, the serum BPA concentration increased as the renal function deteriorated, showing a significant negative association. In a crossover test between PS and cellulose (Ce) dialyzers, the predialysis serum BPA concentration of PS dialyzer users decreased after changing to a Ce dialyzer, and the serum BPA increased again after switching back to PS dialyzers. In patients who were using PS dialyzers, the BPA level significantly increased after a dialysis session. However, in the Ce dialyzer users, the BPA level decreased. Since accumulation of BPA could affect the endocrine or metabolic system of the human body, it is important to perform further investigations on dialysis patients.

In order to fold non-native proteins, chaperonin GroEL undergoes numerous conformational changes and GroES binding in the ATP-dependent reaction cycle. We constructed the real-time three-dimensional-observation system at high resolution using a newly developed fast-scanning atomic force microscope. Using this system, we visualized the GroES binding to and dissociation from individual GroEL with a lifetime of 6 s (k=0.17 s(-1)). We also caught ATP/ADP-induced open-closed conformational changes of individual GroEL in the absence of qGroES and substrate proteins. Namely, the ATP/ADP-bound GroEL can change its conformation 'from closed to open' without additional ATP hydrolysis. Furthermore, the lifetime of open conformation in the presence of ADP ( approximately 1.0 s) was apparently lower than those of ATP and ATP-analogs (2-3 s), meaning that ADP-bound open-form is structurally less stable than ATP-bound open-form. These results indicate that GroEL has at least two distinct open-conformations in the presence of nucleotide; ATP-bound prehydrolysis open-form and ADP-bound open-form, and the ATP hydrolysis in open-form destabilizes its open-conformation and induces the 'from open to closed' conformational change of GroEL.

The degradation of benthiavalicarb-isopropyl (BVI) in soils was studied in the laboratory using two 14C-compounds labeled at the benzene ring and the valine-moiety. The half-life of BVI in two kinds of soils (Ushiku soil and Kakegawa soil) was less than 11 days, under both upland and flooded conditions. Five major degradation products, in addition to 14CO2, were identified. It is presumed that the degradation begins initially with hydrolytic cleavage of the amide bond, followed by several processes resulting in the final production of 14CO2. © Pesticide Science Society of Japan

Three monoamine oxidase (MAO) inhibitors were isolated from Gentiana lutea . Their structures were elucidated to be 3-3″ linked-(2′-hydroxy-4- O -isoprenylchalcone)-(2‴-hydroxy-4″- O -isoprenyldihydrochalcone) ( 1 ), 2-methoxy-3-(1,1′-dimethylallyl)-6a,10a-dihydrobenzo(1,2- c )chroman-6-one and 5-hydroxyflavanone. These compounds, and the hydrolysis product of 1 , displayed competitive inhibitory properties against MAO-B which was more effective than MAO-A.

A method for residue analysis of benthiavalicarb-isopropyl, its diastereomer, and its four degradation products in soil by HPLC was established. The method of extraction under reflux with a mixture of acetone and ammonium chloride solution was optimal for most of the compounds. The limit of quantification in two soils was 0.01 to 0.02 mg/kg for the compounds, and the recovery rates of the compounds from the soils were 74 to 113%. But for the degradation product having an amino group, the rate of recovery from one soil only was good. The level of benthiavalicarb-isopropyl in two upland field soils decreased rapidly.

A method for semi-micro high-performance liquid chromatography (HPLC) has been established for the simultaneous determination of 3alpha-hydroxyglycyrrhetic acid and 3-dehydroglycyrrhetic acid together with glycyrrhizin, glycyrrhetic acid and glycyrrhetic acid mono-glucuronide formed by incubation of glycyrrhizin with rat feces. The analysis was accomplished within 25 min with a TSKgel ODS-80TsQA (150 x 2.0 mm i.d.) column by linear gradient elution using a mobile phase containing aqueous phosphoric acid and acetonitrile at a flow rate of 0.2 ml.min(-1), a thermostatic oven at 25 degrees C, and detection at 254 nm. The detection limits of these compounds were 0.2 pmol per injection (5 microl). The metabolites of glycyrrhizin, by anaerobic or aerobic incubation with rat fecal suspension over 48 h, were determined. Glycyrrhizin was almost completely converted to metabolite glycyrrhetic acid, and metabolites 3alpha-hydroxyglycyrrhetic acid and 3-dehydroglycyrrhetic acid in negligible amounts in anaerobic conditions. However, the metabolic time courses of 3-dehydroglycyrrhetic acid when incubated in aerobic conditions revealed that it apparently continued increasing during the whole incubation period.

Purpose. Parent cyclodextrins are known to accelerate the degradations such as dehydration and isomerization of E-type prostaglandins in neutral and alkaline solutions. The objective of this study was to attempt the stabilization and solubilization of E1-type prostaglandin analogue in aqueous solution by biocompatible cyclodextrin derivatives.

A method for semi-micro high-performance liquid chromatography (HPLC) has been established for the simultaneous determination of glycyrrhizin (GL), glycyrrhetic acid (GA) and glycyrrhetic acid mono-glucuronide (GAMG) in incubation mixtures of rat feces with Shakuyaku-kanzo-to decoction (combination of licorice root and peony root). The analysis could be accomplished within 20 min with a TSKgel ODS-80TsQA (150 x 2.0 mm i.d.) column by linear gradient elution using a mobile phase containing aqueous phosphoric acid and acetonitrile at a flow rate of 0.2 ml x min(-1), a thermostatic oven at 25 degrees C, and detection at 254 nm. The detection limits of these compounds were 0.1-0.85 pmol per injection (5 microl). The concentrations of GL and its metabolites in the incubation mixture after continuous consumption of Shakuyaku-kanzo-to were significantly different compared with those of untreated control. GL-hydrolysis of rat feces was enhanced by pre-consumption of Shakuyaku-kanzo-to.

An ion-pair high-performance liquid chromatographic method for the simultaneous determination of four flavonoids, namely baicalin, wogonoside, baicalein and wogonin, and four berberine-type alkaloids, namely berberine, coptisine, palmatine and jateorrhizine, and glycyrrhizin in Kampo medicines is described. The analysis can be accomplished within 30 min with a Wakosil-II 5C18 HG column by linear gradient elution using a mobile phase containing aqueous phosphoric acid, sodium dodecyl sulfate and acetonitrile at a flow-rate of 1.0 ml x min(-1), a thermostatic oven at 45 degrees C, and detection at 265 nm. The method was applied to quantifying these components in three Kampo decoctions: Oren-gedoku-to, San'o-shashin-to and Hange-shashin-to. The decoctions were diluted with 65% methanol at the final stage because a large quantity of precipitate, mainly from baicalin and berberine, was formed. The within-day relative standard deviations were less than 2.02% (n=10). The recoveries of these compounds were 90.3-102%. The detection limits of these compounds were 0.02-1.96 microM per injection (5 microl).

Five new chromone components were isolated from the gel of Aloe vera leaves. Their structures have been established as 8- C -glucosyl-noreugenin, 4′- O -glucosyl-isoaloeresin DI, 4′– O -glucosyl-isoaloeresin DII, 8- C -glucosyl-(2′- O -cinnamoyl)-7- O -methylaloediol A and 8- C -glucosyl-(2′- O -cinnamoyl)-7- O -methylaloediol B, from spectroscopic studies.

Callus tissue of Aloe barbadensis grown in the dark produced two new tetrahydroanthracene glucosides, 3,4-dihydro-2,4,8,9-tetrahydroxy-6-methyl-1(2H)-anthracenone-4- O - β -d-glucopyranoside and 3,4-dihydro-2-methoxy-4,8,9-trihydroxy-6-methyl-1(2H)-anthracenone-4- O - β -d-glucopyranoside. The effect of light on the formation of tetrahydroanthracene glucosides in the calli is discussed.

The leaves of Aloe vera afforded, together with 10-hydroxyaloins A and B, a new pair of diastereoisomeric C -glucosylanthrones whose structures were established as 8- O -methyl-7-hydroxyaloin A and 8- O -methyl-7-hydroxyaloin B from spectroscopic studies. © 1997 Elsevier Science Ltd. All rights reserved

Altersolanol A, a phytotoxic tetrahydroanthraquinone isolated from the culture broth of Alternaria solani , inhibited the growth of cultured cells of Nicotiana rustica . It acted as a potent stimulator of NADH oxidation in mitochondria isolated from N. rustica cells. The stimulation was also observed in mitochondria in which the respiratory chain was blocked by electron transport inhibitors. Altersolanols acted as electron acceptors in an enzyme preparation of diaphorase. The capacity of altersolanols to act as electron acceptors was A>E>C>B>F>D.

A procedure has been developed for determination of aloesin, 2′- O -feruloylaloesin, aloeresin A, barbaloin, isobarbaloin, aloenin, aloe-emodin, 8- C -glucosyl-7- O -methyl-( S )-aloesol, isoaloeresin D and aloeresin E which are phenolic constituents of aloe. Aloe or commercial aloin was extracted with methanol multiple times, centrifuged and then filtered. Filtrates were analyzed by a reversed-phase high-performance liquid chromatography employing UV-Vis detection (290 nm). The samples were separated with a Wakosil-II 5C18 HG column by linear gradient elution using water-acetonitrile (88:12 to 54:46) as the mobile phase at a flow-rate of 1.0 ml/min. The detection limits of these compounds were 0.04–0.35 ng per injection (5 μ1) and linearity of response existed. Very satisfactory and reproducible results were obtained within 38 min for simultaneous determination of these compounds. This method was applied to determine these compounds in Aloe barbadensis Miller, A. arborescens Miller var. natalensis Berger, A. vera var. chinensis Berger, A. marlothii Berger and A. striata Haw. Two commercial aloins were also analyzed.

Three new chromone components, 8-C-glucosyl-7-O-methyl-(S)-aloesol, isoaloeresin D and aloeresin E were isolated from the leaves of Aloe vera. Their structures have been established from spectroscopic studies; the structures of 8-C-glucosyl-7-O-methyl-(S)-aloesol, isoaloeresin D and aloeresin E were shown to be 8-C-β- d - glucopyranosyl -2-[(S)-2- hydroxy]propyl-7-methoxy-5-methylchromone , 8-C-β- d -[2′-O-(E)-p- coumaroyl]glucopyranosyl -2-[(S)-2- hydroxy]propyl-7-methoxy-5-methylchromone and 8-C-β- d -[2′-O-(E)- cinnamoyl]glucopyranosyl -2-[(S)-2- hydroxy]propyl-7-methoxy-5-methylchromone , respectively. The inhibitory action of these compounds against tyrosine oxidation by mushroom tyrosinase was examined.

An antimicrobial diterpene was isolated from Alpinia galanga in the screening for potentiators of phytochemical antibiotic action. The structure was elucidated by spectral data and identified as (E)-8 beta, 17-epoxylabd-12-ene-15, 16-dial (1). Diterpene 1 synergistically enhanced the antifungal activity of quercetin and chalcone against Candida albicans. Antifungal activity of 1 was reversed by unsaturated fatty acids. Protoplasts of C. albicans were lysed by 1. These results suggest that antifungal activity of 1 is due to a change of membrane permeability arising from membrane lipid alternation.

Isoaffinetin (5,7,3',4',5'-pentahydroxyflavone-6-C-glucoside) was isolated from Manilkara indica as a potent inhibitor of lens aldose reductase by bioassay-directed fractionation. This C-glucosyl flavone showed specific inhibition against aldose reductases (rat lens, porcine lens and recombinant human) with no inhibition against aldehyde reductase and NADH oxidase. Kinetic analysis showed that isoaffinetin exhibited uncompetitive inhibition against both dl-glyceraldehyde and NADPH. A structure-activity relationship study revealed that the increasing number of hydroxy groups in the B-ring contributes to the increase in aldose reductase inhibition by C-glucosyl flavones.

An active glycoprotein fraction containing 58 % protein was isolated from Aloe vera gel by precipitation with 55 % ammonium sulfate followed by gel permeation using DEAE-Sephacel A-25, Sepharose 6B and Sephadex G-50 columns in a yield of 3 x 10 -3 %. The glycoprotein fraction showed a single band corresponding to a subunit of verectin at the same position when stained with both Coomassie brilliant blue and periodic acid-Schiff reagents on 18 % SDS-PAGE. The molecular weight (14 kDa) was confirmed by Sephadex G-50 column chromatography. The glycoprotein fraction showed a radical scavenging activity against superoxide anion generated by the xanthine-xanthine oxidase system as well as inhibition of cyclooxygenase-2 and reduction of thromboxane A 2 synthase level in vitro.

AIM: To investigate the anti-oxidative and anti-fibrotic effects of aloe vera in patients with liver fibrosis. METHODS: Aloe vera high molecular weight fractions (AHM) were processed by patented hyper-dry system in combination of freeze-dry technique with microwave and far infrared-ray radiation. Fifteen healthy volunteers as the control group and 40 patients were included. The patients were randomly subdivided into two equal groups: the conventional group was treated with placebo (starch), and AHM group was treated with 0.15 gm/d AHM, both for 12 consecutive weeks. The patients were investigated before and after treatment. Serum activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), hyaluronic acid (HA), transforming growth factor-β (TGF-β) and matrixmetalloproteinase-2 (MMP-2) were determined. The reduced glutathione (GSH) and malondialdehyde (MDA) levels in liver were assayed and the expression of hepatic α-smooth muscle actin (α-SMA) was identified by immunohistochemistry. RESULTS: At the start of the study, the hematoxylin and eosin staining revealed fibro-proliferated bile ductules, thick fibrous septa and dense inflammatory cellular infiltration in the patients before treatment. The use of AHM for 12 wk significantly ameliorated the fibrosis, inhibited the inflammation, and resulted in minimal infiltration and minimal fibrosis compared to the conventional group. The enzyme activities of the liver (ALT, AST and ALP) were attenuated after treatment in both groups, and the decrease in the AHM group was more significant as compared with the conventional group. Similar to the AST, the MDA levels were significantly higher before treatment, and were attenuated after treatment in both groups. In contrast, the hepatic glutathione content in the patients were decreased significantly in the AHM group compared to the controls. The serum levels of the fibrosis markers (HA, TGF-β and MMP-2) were also reduced significantly after treatment. The expression of α-SMA was modified in patients before and after treatment as compared with the normal controls. In the conventional group, there was only thin and incomplete parenchymal α-SMA positive septum joining the thickened centrilobular veins, while in the AHM group, few α-SMA positive cells were present in sinusoid and lobule after treatment. CONCLUSION: Oral supplementation with AHM could be helpful in alleviating the fibrosis and inflammation of hepatic fibrosis patients.

The present study was undertaken to elucidate the effects of sodium tanshinone VI 1-phenolate (1), 1'-O-hydrogen succinyltanshinone VI 1-O-hydrogen succinate (2), and disodium 1'-O-succinyltanshinone VI 1-O-succinate (3), water-soluble derivatives of tanshinone VI, on post-hypoxic contractile recovery of isolated perfused rat hearts. The effects were compared with those of tanshinone VI as tested previously. The hearts were perfused for 20 min under hypoxic conditions, followed by 45 min reoxygenation, and their cardiac performance was determined. Changes in tissue sodium, potassium, calcium, and magnesium contents after reoxygenation, and release of creatine kinase and purines and bases (ATP metabolites) during hypoxia/reoxygenation were also examined. The derivatives were dissolved in a Krebs-Henseleit buffer and administered at concentrations of 42 nM into the buffer. Hypoxia/reoxygenation resulted in slight recovery of cardiac contractile force, significant alterations in tissue ion concentrations, and pronounced release of creatine kinase and ATP metabolites, suggesting hypoxia/reoxygenation-induced functional and morphological damage. The tanshinone VI derivatives improved post-hypoxic contractile recovery, which was associated with restoration of tissue ionic concentrations, and diminishment of the release of creatine kinase and ATP metabolites from the hypoxic/reoxygenated hearts. The efficacy of these compounds was similar to that of tanshinone VI. The results suggest that water-soluble tanshinone VI derivatives, like tanshinone VI itself, are beneficial for hypoxia/reoxygenation injury.

A reversed-phase high-performance liquid chromatographic method for the determination of carnosic acid and carnosol, phenolic diterpenes, which showed high antioxidative effect, is established. The analysis can be accomplished within 15 min under isocratic conditions with 0.1% phosphoric acid-60% acetonitrile as the mobile phase at a flow-rate of 1.0 ml/min, with detection at 230 nm. The detection limits of carnosic acid and carnosol are 0.104 and 0.521 ng per injection, respectively. This experimental system permits a good separation and quantification of these phenolic diterpenes in the leaves of Rosmarinus officinalis L. and Salvia officinalis L.

The in vitro and in vivo skin permeability of 16 drugs with a wide span of lipophilicity (log P ranging from -0.95 to 4.40) was evaluated with an ethanol/panasate 800 (tricaprylin) (40/60) system as a lipophilic vehicle. The ethanol/panasate 800 (40/60) binary vehicle remarkably improved the in vitro skin permeability of the drugs across excised hairless mouse skin compared with ethanol or panasate 800 as single vehicles. The in vivo skin permeability of the large majority of the drugs across abdominal rat skin also showed high permeation rates and short lag times. The relationship between lipophilicity and skin permeability of the drugs from the ethanol/panasate 800 (40/60) binary vehicle indicated parabolic shapes with their peaks at much greater hydrophilic range (log P: -0.88 for in vitro, -0.83 for in vivo) compared with other past references (log P: 2-3). The results suggest that the lipophilicity of a drug is a main factor for prediction of the skin permeability of the drug and that the ethanol/panasate 800 (40/60) lipophilic binary vehicle would be a good candidate as a vehicle for future clinical application of hydrophilic drugs.

This study was performed to determine the active constituents of the root of Panax ginseng C. A. Mayer in the amelioration of ethanol-induced impediment of brain growth in the neonatal stage. To establish an animal model of the brain growth impediment caused by ethanol, ethanol (6 g/kg s.c.) was administered to rat pups on postnatal day 6, which corresponded to the third trimester of pregnancy for humans. Brain weight, especially cerebellar weight, was significantly reduced in the ethanol-exposed pups. In contrast, neither separation from dams nor pentobarbital treatment affected brain weight. A saponin fraction of ginseng extract prevented this ethanol-induced reduction of brain weight. Some ginseng saponins including ginsenosides Rg1, Rb2, Rd, Rf and Re effected stimulated a potent recovery of cerebellum growth in this animal model.

The in vitro skin permeability of 16 drugs with a wide range of lipophilicity (log P ; -0.95-4.40) was evaluated by the use of an ethanol/water (60/40) binary vehicle with of without lauric acid as a permeation enhancer. The enhancing effect by the addition of lauric acid to the ethanol/water (60/40) binary vehicle could be observed from the aspect of both permeation rate and lag time. The permeation rate increased with an increase in the hydrophilicity of the drugs. It was considered that lauric acid exerts an effect on both the polar and nonpolar regions of lipids of the stratum corneum, and the cooperative interaction of ethanol and lauric acid increases the participation of the polar pathway of drugs. The relationship between lipophilicity and skin permeability of the drugs from the ethanol/water (60/40) binary vehicle with lauric acid showed a parabolic shape, with its peak at a more hydrophilic range (log P ; 0.19) compared with other past references (log P ; 2-3). The ethanol/water (60/40) binary vehicle with lauric acid appears to be a good candidate as a vehicle for transdermal therapeutic systems for hydrophilic drugs.

A high-performance liquid chromatographic model for the simultaneous determination of altersolanol A–F by using a reversed-phase column with an acetonitrile-water gradient elution system is described. The analysis can be completed within 13 min, the detection limits are 0.2–0.5 pmol per injection (5 μl) and the relative standard deviations are 0.90–1.34%. The method was applied satisfactorily to the determination of altersolanols in culture media of a strain of Alternaria solani without any prepurification.

The ginseng tetrapeptide H-L-Val-γ-D-Gly-D-Arg-Gly-OH (1) was synthesized using a solution-phase methodology, and its solution conformation in dimethyl sulfoxide (DMSO)-d 6 was determined by a combination of NMR and computational techniques with the DADAS90 program. This compound has a rigid backbone based on three intramolecular hydrogen bonds between D-Arg NH and L-Val CO, between Gly NH and D-Glu CO, and between Gly NH and Gly CO.

The antimicrobial mechanism of altersolanol A was studied by using Pseudomonas aeruginosa IFO 3080. This reduced anthraquinone antibiotic inhibited the incorporation of radioactive precursors into macromolecules (DNA, RNA, and Protein) in whole cells nonspecifically. It increased respiration of P. aeruginosa cells, though it had no effect on proton condution. Oxidation of NADH in the membrane fraction isolated from P. aeruginosa was enhanced by altersolanol A. Furthermore, altersolanol A stimulated the oxidation of NADH by the enzyme preparation of cytochrome c reductase in the absence of cytochrome c. Altersolanol A seemed to interfere with the respiratory chain in the bacterial membrane as an electron acceptor.

A sensitive, quantitative procedure was developed for the simultaneous determination of tanshinone IIA and I, cryptotanshinone and dihydrotanshinone I by high-performance liquid chromatography (HPLC) employing a normal-phase column. In addition, a method for the determination of the polar tanshinones tanshinone V and VI by reversed-phase HPLC, an HPLC method for ferruginol and a qualitative procedure for other tanshinones were also developed. These methods were applied to identify abietane-type compounds in various tissues of Salvia miltiorrhiza Bung, including cultured cells, fresh roots and regenerated plant roots.

The complex dielectric constant of the high-frequency dielectric relaxation of sodium polystyrene sulfonate in aqueous solutions was measured over wide ranges on degree of polymerization, N, and polymer concentration, C p , from the dilute to the semidilute region. The crossover behavior of the high-frequency relaxation is ascribable to that of the correlation length, ξ, which shows different C p dependences in the dilute and semidilute regions, respectively

Newly developed fast-scanning atomic force microscopy (AFM) allows the dissection of molecular events such as DNA–enzyme reactions at the single-molecule level. With this novel technology, a model is proposed of the DNA cleavage reaction by a type IIP restriction endonuclease ApaI. Detailed analyses revealed that ApaI bound to DNA as a dimer and slid along DNA in a one-dimensional diffusion manner. When it encountered a specific DNA sequence, the enzyme halted for a moment to digest the DNA. Immediately after digestion, the ApaI dimer separated into two monomers, each of which remained on the DNA end and then dissociated from the DNA end. Thus, fast-scanning AFM is a powerful tool to aid the understanding of protein structures and dynamics in biological reactions at the single-molecule level in sub-seconds.