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Multiple myeloma (MM) belongs to hematological malignancy that derives pro-survival/proliferative signals from the bone marrow niche, and its incidence is increasing worldwide. Butyrate, a well-known epigenetic histone deacetylase inhibitor, reduces the viability of MM cells, induces MM cell apoptosis via transcriptional activation of p21. Inpresent review we discuss butyrate as a potential therapeutic drug for MM and case reports of MM, acute leukemia, malignant lymphoma and glioma patients.

Our health span is to a great extent epigenetically determined by diets, gut environment, lifestyle, and various other environmental factors. Health benefits of long-term ingestion of aloe vera gel with immune modulation of the intestinal microbiota were discussed on congenital heart disease and heart failure. In this review, we demonstrated prophylaxes roles of successive ingestion of aloe vera juice on heart failure. Furthermore we presented three case reports on heart failure patients with aloe vera juice adjuvant.

The mechanism of how gut microbiota is involved in the host inflammatory processes that influence human immunodeficiency virus (HIV) replication and disease progression in HIV is elucidated in present review.

We discussed evident effects of aloe-emodin and aloe polysaccharide on the therapy of viral infections and epigenetic factor butyrate that is involved in latency and reactivation in viral infection. Furthermore we presented eight case reports of Kampo medicine with Aloe vera juice (AVJ) to virus infections. Kampo medicine with AVJ-successive ingestion demonstrates to be suitably personalized medicine for virus infections and improves gut-microbial ecosystem in host. The potential Kampo medicine with AVJ may provide fresh insights as an anti-virus medicine for homeostatic regulation in host.

We present two case reports: beneficial roles of successive ingestion of aloe vera juice for medical treatment of steroid-sensitive nephrotic kidney syndrome in child.

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.

Our healthspan is to a great extent epigenetically determined by diets, lifestyle, and various other environmental factors. Gut microbiota has been proven to be the major player in maintaining human health. Our previous report described health benefits of long-term ingestion of aloe vera gel through the modification of the intestinal microbiota. In the present review, we broadly cover the topics of microbiota and aloe vera gel ingestion related to attenuation of reactive oxygen species, prevention of cardiovascular disorders, effects of life-prolonging calorie restriction. Additional data are summarized on prophylactic actions of fermented butyrate, and aloe-emodin related components against obesity. Prophylactic actions of aloe vera gel on healthy aging, skin photo-aging, and the viability and lifespan in Drosophila melanogaster are discussed in relation to the properties of butyrate and its potential effects involved in health and disease.

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.

Based on the standpoint of natural treasure: Aloe vera and bee-products, we presented the following information: (i) cardiovascular activity, (ii) usefulness to benign prostatic hyperplasia, chronic prostatitis, and menopausal syndromes, (iii) enhancement of muscle performance in athletes, (iv) others in vivo therapeutic effects, (v) side effects, and (vi) honey, (vii) response of chronic fatigue subjects to the ingestion of Aloe vera juice (AVJ) with or without bee-products (propolis and pollen), and case reports, (viii) future prospective, and (ix) summary. The efficacies of AVJ ingestion with bee-products supplement were demonstrated in questionnaire assessments: response of AVJ with bee products ingestion to chronic fatigue subjects. The results provided new possibilities for incorporation of bee-products with AVJ as quality of life-food stuffs. Futhermore, case reports of AVJ with bee-products to patients in metabolic syndrome and inflammation around the under-rectum in ulcerative colitis were demonstrated. Ingestion of Aloe vera juice with bee-products supplement could provide the noteworthy improvement.

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.

The efficacy of Aloe vera has been known for various health and disease-related conditions. Butyrate, a well-known epigenetic histone deacetylase inhibitor, is causally implicated in tumorigenesis and tumor angiogenesis. Butyrate was shown to induce cell cycle arrest, differentiation, and apoptosis in a variety of cancer cells. Dietary factors, microbiota composition, and microbiota metabolism are intimately intertwined in a complex network, highlighting the importance of intestinal functions for health maintenance. In the present review, we discuss aloe vera fermentation as a tumor-suppressive process generating microbial-derived butyrate.

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.

Symbiotic effect of butyrate-producing endophytic microbiota and Aloe vera gel containing non-digestible carbohydrates was discussed on slowing ageing design: butyrate efficacy for insulin sensitivity, sirtuin activation through histone deacetylase inhibitors in vitro study. Possible putative efficacy of butyrate fermented by endophytic microbiota for insulin sensitivity on glucose homeostasis is discussed.

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.

L-Arginine is one of the most metabolically versatile amino acids. Several in vitro/in vivo experiments have indicated that exogenous L-arginine intake has multiple beneficial pharmacological and pharmaco-kinetic effects. Such effects include reduction in the risk of vascular and heart diseases and chronic fatigue syndrome. The effects of a dietary supplementation of aloe vera juice with L-arginine to chronic fatigue syndrome were demonstrated and a questionnaire was given to adult subjects. The questionnaire included 10 points regarding their mental, circulatory and muscle functions. Table 1 and 2 summarize the most noteworthy information on supplementation of aloe vera juice with L-arginine. The demonstrated benefits of aloe vera juice with L-arginine show promises to chronic fatigue syndrome in adult subjects.

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.

Coenzyme Q (CoQ10) is a major antioxidant principle found in human body which plays a vital role in maintaining several biochemical pathways of body. Deficiency of CoQ10 in body leads to several potential disorders in oxido-reductive reactions of electron transport chain. Present review offers preliminary insight into using dietary supplements, aloe vera juice with CoQ10, to support and optimize quality of life in the people suffering from muscle performance. A questionnaire on aloe vera juice with CoQ10 supplement was given to adult subjects. The questionnaire included 10 points regarding their mental, circulatory, and muscular functions, and the scoring was recorded from top six conditions in Table 1 and 2. Table 2 summarizes the noteworthy information of aloe vera juice with CoQ10 supplement. The high molecular weight and lipophilicity of CoQ10, making its poor water solubility and leading to low systemic availability, may be reduced by supplementation of aloe vera juice with CoQ10. Possible prophylaxes of aloe vera juice with CoQ10 supplement to enhance muscle performance are fully expected.

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.

An improved unroofing method enabled the cantilever of an atomic force microscope (AFM) to reach directly into a cell to visualize the intracellular cytoskeletal actin filaments, microtubules, clathrin coats and caveolae in phosphate-buffered saline (PBS) at a higher resolution than conventional electron microscopy. All of the actin filaments clearly exhibited a short periodicity of approximately 5–6 nm, which was derived from globular actins linked to each other to form filaments, as well as a long helical periodicity. The polarity of the actin filaments appeared to be determined by the shape of the periodic striations. Microtubules were identified based on their thickness. Clathrin coats and caveolae were observed on the cytoplasmic surface of cell membranes. The area containing clathrin molecules and their terminal domains was directly visualized. Characteristic ridge structures located at the surface of the caveolae were observed at high resolution, similar to those observed with electron microscopy (EM). Overall, unroofing allowed intracellular AFM imaging in a liquid environment with a level of quality equivalent or superior to that of EM. Thus, AFMs are anticipated to provide cutting-edge findings in cell biology and histology.

Homophilic binding of E-cadherins through their ectodomains is fundamental to epithelial cell-cell adhesion. Despite this, E-cadherin ectodomains have evolved differently in the vertebrate and insect lineages. Of the five rod-like, tandemly aligned extracellular cadherin domains of vertebrate E-cadherin, the tip extracellular cadherin domain plays a pivotal role in binding interactions. Comparatively, the six consecutive N-terminal extracellular cadherin domains of Drosophila E-cadherin, DE-cadherin (also known as Shotgun), can mediate adhesion; however, the underlying mechanism is unknown. Here, we report atomic force microscopy imaging of DE-cadherin extracellular cadherin domains. We identified a tightly folded globular structure formed by the four N-terminal-most extracellular cadherin domains stabilized by the subsequent two extracellular cadherin domains. Analysis of hybrid cadherins from different insects indicated that the E-cadherin globular portion is associated with determining homophilic binding specificity. The second to fourth extracellular cadherin domains were identified as the minimal portion capable of mediating exclusive homophilic binding specificity. Our findings suggest that the N-terminal-most four extracellular cadherin domains of insect E-cadherin are functionally comparable with the N-terminal-most single extracellular cadherin domain of vertebrate E-cadherin, but that their mechanisms might significantly differ. This work illuminates the divergence of structural strategies for E-cadherin homophilic binding among bilaterians.

Homophilic binding of E-cadherins through their ectodomains is fundamental to epithelial cell-cell adhesion. Despite this, E-cadherin ectodomains have evolved differently in the vertebrate and insect lineages. Of the five rod-like, tandemly aligned extracellular cadherin domains of vertebrate E-cadherin, the tip extracellular cadherin domain plays a pivotal role in binding interactions. Comparatively, the six consecutive N-terminal extracellular cadherin domains of Drosophila E-cadherin, DE-cadherin (also known as Shotgun), can mediate adhesion; however, the underlying mechanism is unknown. Here, we report atomic force microscopy imaging of DE-cadherin extracellular cadherin domains. We identified a tightly folded globular structure formed by the four N-terminal-most extracellular cadherin domains stabilized by the subsequent two extracellular cadherin domains. Analysis of hybrid cadherins from different insects indicated that the E-cadherin globular portion is associated with determining homophilic binding specificity. The second to fourth extracellular cadherin domains were identified as the minimal portion capable of mediating exclusive homophilic binding specificity. Our findings suggest that the N-terminal-most four extracellular cadherin domains of insect E-cadherin are functionally comparable with the N-terminal-most single extracellular cadherin domain of vertebrate E-cadherin, but that their mechanisms might significantly differ. This work illuminates the divergence of structural strategies for E-cadherin homophilic binding among bilaterians.

Actin filaments, the actin-myosin complex and the actin-tropomyosin complex were observed by a tip-scan atomic force microscope (AFM), which was recently developed by Olympus as the AFM part of a correlative microscope. This newly developed AFM uses cantilevers of similar size as stage-scan AFMs to improve substantially the spatial and temporal resolution. Such an approach has previously never been possible by a tip-scan system, in which a cantilever moves in the x, y and z directions. We evaluated the performance of this developed tip-scan AFM by observing the molecular structure of actin filaments and the actin-tropomyosin complex. In the image of the actin filament, the molecular interval of the actin subunits (∼5.5 nm) was clearly observed as stripes. From the shape of the stripes, the polarity of the actin filament was directly determined and the results were consistent with the polarity determined by myosin binding. In the image of the actin-tropomyosin complex, each tropomyosin molecule (∼2 nm in diameter) on the actin filament was directly observed without averaging images of different molecules. Each tropomyosin molecule on the actin filament has never been directly observed by AFM or electron microscopy. Thus, our developed tip-scan AFM offers significant potential in observing purified proteins and cellular structures at nanometer resolution. Current results represent an important step in the development of a new correlative microscope to observe nm-order structures at an acceptable frame rate (∼10 s/frame) by AFM at the position indicated by the fluorescent dye observed under a light microscope.

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.