Antitumor activity and underlying mechamisms of ganopoly, the refined polysaccharides extracted from ganodrema lucidum, in mice

Ganopoly is an aqueous polysaccharide fraction extracted from G. lucidum by patented biochemical technique and has been marketed as an over-the-counter product for chronic diseases including cancer and hepatopathy in many Asian countries. This study was undertaken to explore the anti-tumour effect and the underlying mechanisms of Ganopoly in mice and human tumor cell lines. The maximum tolerateddose (MTD) of Ganopoly in mice was estimated to be 100 mg/kg from a pilot study. Treatment of mice with oral Ganopoly for 10 days significantly reduced the tumour weight of sarcoma-180 in a dose-dependent manner, with inhibition rates of 32.3, 48.2 and 84.9% and growth delays of 1.5, 3.5, and 13.1 days at 20, 50, and 100 mg/kg, respectively. Incubation of Ganopoly at 0.05-1.0 mg/ml for 48 hours showed little or negligible cytotoxicity against human tumor CaSki, SiHa, Hep3B, HepG2, HCT116, HT29, and MCF7 cells in vitro. In contrast, 10 mg/ml of Ganopoly caused significant cytotoxicity in all tumour cells tested except MCF7, with marked apoptotic effects observed in CaSki, HepG2, and HCT116 cells, as indicated by nuclear staining and DNA fragmentation. In addition, Ganopoly enhanced concanavalin A-stimulated proliferation of murine splenocytes by 35.3% at 10 mg/ml, and stimulated the production of nitric oxide in thioglycollate-primed murine peritoneal macrophages in a concentration-dependent manner over 0.05-10 mg/ml. Addition of Ganopoly at 1 mg/ml to murine peritoneal macrophages also potentiated lipopolysaccharide-induced nitric oxide production by 64.2%. Treatment of healthy mice or mice bearing sarsoma-180 with oral Ganopoly over 20-100 mg/kg for 7 day significantly increased the expression of both TNF-α and IFN-γ (at both mRNA and protein levels) in splenocytes in a dose-dependent manner. Moreover, treatment of Ganopoly over 20-100 mg/kg significantly increased cytotoxic T lymphocyte cytotoxicity and NK activity in mice. The overall findings indicated that Ganopoly had antitumor activity with a broad spectrum of immuno-modulating activities and may represent a novel promising immunotherapeutic agent in cancer treatment.

Immunostimulatory polysaccharides from Chlorella pyrenoidosa. A new galactofuranan. Measurement of molecular weight and molecular weight dispersion by DOSY NMR.

Fractionation of the hot water extract of Chlorella pyrenoidosa was performed using a combination of ethanol precipitation, size exclusion chromatography, and anion exchange chromatography. One fraction contained a new polysaccharide, and this compound was shown to be a 1→2-linked β-d-galactofuranan from its 1D and 2D 1H and 13C NMR spectra, with a molecular weight of 15 kDa from DOSY NMR measurements. A number of other fractions were shown to have the same repeating unit as the previously identified arabinogalactan. However, arabinogalactans from different fractions were shown by DOSY NMR to have different molecular weights, which ranged from 27 to 1020 kDa. Agreement with molecular weights measured for some of these fractions by SEC-MALS was very good, further confirming the relationship established by Viel et al. between molecular weights of neutral polysaccharides and self-diffusion coefficients. The smaller molecular weight polysaccharides, the galactofuranan and the 27 and 50 kDa arabinogalactans, were shown to be close to monodisperse by analysis of the distributions of the self-diffusion coefficients for the polymers. The larger arabinogalactans had considerable variation in their molecular weights (188 ± 109 kDa and 1020 ± 370 kDa). Only the two larger arabinogalactans showed immunostimulatory activity.

Aspects of the production of viscous capsular material by the yoghurt starter, streptococcus thermophilus

The technology of modern fermented milk production is not complicated and relies largely on the characteristics of the microorganisms used in its manufacture. Biochemical substances excreted by the starter cultures contribute to the chemical, physical and organoleptic properties of cultured milks. Chemical and organoleptic properties of yoghurt starter cultures have been widely studied over several decades. Conversely the biosynthetic processes and genetic control of the production of viscous extracellular material (slime) by selected thermophillic streptococci is still insufficiently understood. This study attempted to elucidate physiological aspects and the genetic control of slime production. An attempt to chemically induce ropiness was also preformed. Twenty strains of Gram positive, thermo-tolerant, milk dotting, catalase negative cocci were collected from a variety of sources. All strains were identified as Streptococcus thermophilus. Four of the isolates were identified as capable of producing an extracellular, ‘ropy’ capsular material. A negative staining method for highlighting capsular material under light microscopy was described. Ropy isolates displayed thick capsular zones of between 6-8 μm. The isolates graded as non-ropy produced only small capsular zones (less than 2 μm); two variants displayed no capsular material. Instability of the ropy phenotype during subculture and prolonged storage was described for all four ropy isolates at varied temperatures. Instability during transfer was reported as moderate with a loss of no more than 45% of ropy colonies after 15 subcultures at 48°C A significant increase in instability, during transfer, associated with an increase in incubation temperature (37-48°C) was also reported. Prolonged storage of ropy variants over ten days resulted in a drop in the number of ropy colonies. The loss was minimal when cultures were stored at 8°C, but excessive (approaching 100%) at 37°C This suggested the presence of capsular degradative substances. Analysis of the plasmid profiles of 20 strains identified only two strains harboured plasmid DNA. All plasmids were small, less than 23kilobases, and each strain possessed a single plasmid species. Only one ropy strain contained plasmid DNA that was shown, with the aid of curing experiments, not to be linked to production of the ropy phenotype. The amino acid analogue p-fluoro-DL-phenylalanine was unsuccessful in generating ropy colonies from non-ropy variants of Streptococcus thermophilus at low concentrations. Some technological considerations for the use of ropy variants of Streptococcus thermophilus in yoghurt starter cultures were made.

A self-supported, flexible, binder-free pseudo-supercapacitor electrode material with high capacitance and cycling stability from hollow, capsular polypyrrole fibers

Flexible energy devices with high performance and long-term stability are highly promising for applications in portable electronics, but remain challenging to develop. As an electrode material for pseudo-supercapacitors, conducting polymers typically show higher energy storage ability over carbon materials and larger conductivity than transition-metal oxides. However, conducting polymer-based supercapacitors often have poor cycling stability, attributable to the structural rupture caused by the large volume contrast between doping and de-doping states, which has been the main obstacle to their practical applications. Herein, we report a simple method to prepare a flexible, binder-free, self-supported polypyrrole (PPy) supercapacitor electrode with high cycling stability through using novel, hollow PPy nanofibers with porous capsular walls as a film-forming material. The unique fiber structure and capsular walls provide the PPy film with enough free-space to adapt to volume variation during doping/de-doping, leading to super-high cycling stability (capacitance retention > 90% after 11000 charge-discharge cycles at a high current density of 10 A g^-1) and high rate capability (capacitance retention ∼ 82.1% at a current density in the range of 0.25-10 A g^-1).

Capsular polypyrrole hollow nanofibers: an efficient recyclable adsorbent for hexavalent chromium removal

Capsular polypyrrole hollow nanofibers (PPy-HNFs) were fabricated via in situ polymerization of pyrrole on an organic-inorganic template, followed by acid etching. Their application in removing hexavalent chromium (Cr(vi)) from aqueous solution was then investigated. The morphologies of the capsular PPy-HNFs were studied by both scanning electron microscopy (SEM) and transmission electron microscopy (TEM), which showed that the PPy-HNFs had a capsular structure in the walls of hollow nanofibers. Fourier transform infrared (FTIR) spectroscopy and X-ray photoelectron spectroscopy (XPS) data confirmed the adsorption of Cr on capsular PPy-HNFs. The adsorption capacity increased with reduced pH of the initial solution and the adsorption process can be described using the pseudo-second-order model. These capsular PPy-HNFs showed a high Cr(vi) adsorption capacity up to 839.3 mg g-1. This adsorption capacity was largely retained even after five adsorption/desorption cycles. Electrostatic attraction between Cr and PPy-HNFs was studied using a proposed adsorption mechanism. The capsular PPy-HNFs formed a flexible membrane, which allowed easy handling during application. This study has demonstrated the possibilities of using this capsular PPy-HNF membrane for heavy metal removal from aqueous solution.

Isolating conifer DNA: a superior polysaccharide elimination method

Genomic DNA was isolated from frozen needles of maturePinus radiata clones using a modified extraction technique incorporating cetyltrimethylammonium bromide (CTAB) for cell lysis. A high sodium chloride concentration (2 M) was used at 2 stages of the extraction procedure to eradicate polysaccharides, yielding pure genomic DNA suitable for restriction enzyme digestion and PCR amplification. Extractions were scaled down to suit 1.5-mL Eppendorf tubes, allowing easier handling and enhanced sterility.

Diet, nutrition and the prevention of excess weight gain and obesity

Objective: To review the evidence on the diet and nutrition causes of obesity and to recommend strategies to reduce obesity prevalence.
Design: The evidence for potential aetiological factors and strategies to reduce obesity prevalence was reviewed, and recommendations for public health action, population nutrition goals and further research were made.
Results: Protective factors against obesity were considered to be: regular physical activity (convincing); a high intake of dietary non-starch polysaccharides (NSP)/fibre (convincing); supportive home and school environments for children (probable); and breastfeeding (probable). Risk factors for obesity were considered to be sedentary lifestyles (convincing); a high intake of energy-dense, micronutrient-poor foods (convincing); heavy marketing of energy-dense foods and fast food outlets (probable); sugar-sweetened soft drinks and fruit juices (probable); adverse social and economic conditions—developed countries, especially in women (probable).
A broad range of strategies were recommended to reduce obesity prevalence including: influencing the food supply to make healthy choices easier; reducing the marketing of energy dense foods and beverages to children; influencing urban environments and transport systems to promote physical activity; developing community-wide programmes in multiple settings; increased communications about healthy eating and physical activity; and improved health services to promote breastfeeding and manage currently overweight or obese people.
Conclusions: The increasing prevalence of obesity is a major health threat in both low- and high income countries. Comprehensive programmes will be needed to turn the epidemic around.

Dietary assimilation and the digestive strategy of the omnivorous anomuran land crab Birgus latro (Coenobitidae).

On Christmas Island, Indian Ocean, the diet of robber crabs, Birgus latro (Linnaeus) was generally high in fat, storage polysaccharides or protein and largely comprised fruits, seeds, nuts and animal material. The plant items also contained significant amounts of hemicellulose and cellulose. In laboratory feeding trials, crabs had similar intakes of dry matter when fed artificial diets high in either fat or storage polysaccharide, but intake was lower on a high protein diet. Assimilation coefficients of dry matter (69–74%), carbon (72–81%), nitrogen (76–100%), lipid (71–96%) and storage polysaccharide (89–99%) were high on all three diets. B. latro also assimilated significant amounts of the chitin ingested in the high protein diet ( 93%) and hemicellulose (49.6–65%) and cellulose (16–53%) from the high carbohydrate and high fat diets. This is consistent with the presence of chitinase, hemicellulase and cellulase enzymes in the digestive tract of B. latro. The mean retention time (27.2 h) for a dietary particle marker (⁵⁷Co-labelled microspheres) was longer than measured in leaf-eating land crabs. The feeding strategy of B. latro involves the selection of highly digestible and nutrient-rich plant and animal material and retention of the digesta for a period long enough to allow extensive exploitation of storage carbohydrates, lipids, protein and significant amounts of structural carbohydrates (hemicellulose, cellulose and chitin).

Monitoring of immune responses to a herbal immuno-modulator in patients with advanced colorectal cancer.

Many herbal medicines are widely used as immuno-modulators in Asian countries. Ganoderma lucidum (Lingzhi) is one of the most commonly used herbs in Asia and preclinical studies have established that the polysaccharide fractions of G. lucidum have potent immuno-modulating effects. However, clinical evidence for this is scanty. The present open-labeled study aimed to evaluate the effects of G. lucidum polysaccharides on selected immune functions in patients with advanced colorectal cancer. Forty-seven patients were enrolled and treated with oral G. lucidum at 5.4 g/day for 12 weeks. Selected immune parameters were monitored using various immunological methods throughout the study. In 41 assessable cancer patients, treatment with G. lucidum tended to increase mitogenic reactivity to phytohemagglutinin, counts of CD3, CD4, CD8 and CD56 lymphocytes, plasma concentrations of interleukin (IL)-2, IL-6 and interferon (IFN)-γ, and NK activity, whereas plasma concentrations of IL-1 and tumor necrosis factor (TNF)-α were decreased. For all of these parameters, no statistical significance was observed when a comparison was conducted between baseline and those values after a 12-week treatment with G. lucidum. The changes of IL-1 were correlated with those for IL-6, IFN-γ, CD3, CD4, CD8 and NK activity (p < 0.05) and IL-2 changes were correlated with those for IL-6, CD8 and NK activity. The results indicate that G. lucidum may have potential immuno-modulating effect in patients with advanced colorectal cancer. Further studies are needed to explore the benefits and safety of G. lucidum in cancer patients.

Amylase and glucosidase enzyme inhibitory activity of ginger (Zingiber officinale Roscoe) an in vitro study

The prevalence of type 2 diabetes has reached to an epidemic proportion in Sri Lanka. The need for achieving better control of blood glucose level has been evident in diabetes management. However it is not easy to achieve this goal in a large proportion of patients. This is partly due to limitations of currently available pharmacological agents which stimulate research on novel anti-diabetic agents with different mechanisms. Digestive enzymes have been targeted as potential avenues for modulation of blood glucose concentration through inhibition of the enzymatic breakdown of complex carbohydrates to meal derived glucose absorption. Acarbose is a widely used oral anti-diabetic drug which inhibits the α-glucosidase, enzyme responsible for breaking down of disaccharides and polysaccharides into glucose. Many herbal extracts have been found to posses similar inhibitory effects. Ginger (Zingiber officinale Roscoe) has developed a reputation in treatment of several diseases. In vitro enzymic inhibitory effect of ginger was investigated in this study. Enzymes α -amylase and α -glucosidase treated with either Acarbose or ginger extract were allowed to react with cooked rice and percentages of glucose content were measured. The glucosidase and amylase activities on the rice were inhibited by addition of ginger cause significant reduction in glucose percentages (36.86± 1.05 to 26.87± 2.17, P<0.05 and 49.04±0.65 to 35.35±2.22, P<0.05) which showed comparable results with Acarbose on glucosidase activity (36.86± 1.05 to, 27.8±1.32 P<0.05). Results of the study indicates ginger as a potential plant based amylase and glucosidase inhibitor in carbohydrate digestion but usage in glycaemic control in human has to be investigated further.

Oat beta-glucan lowers total and LDL-cholesterol

Several soluble polysaccharides have been shown to have cholesterol-lowering properties and to have a role in prevention of heart disease. Major sources of one such polysaccharide (beta-glucan) are oats and barley. The aim of this study was to examine the effects on plasma lipid concentrations when beta-glucan derived from a fractionated oat preparation was consumed by people with elevated plasma lipids. A single-blind, crossover design compared plasma cholesterol, triglycerides, high density lipoproteins and low density lipoproteins (LDLs) in 14 people; in the order of low, high and low beta-glucan supplemented diets, each of three weeks duration. For the high beta-glucan diet, an average intake of 7 g per day was consumed from cereal, muffins and bread. The background diet remained relatively constant over the three test periods. Differences during the interventions were calculated by one-way repeated measures analysis of variance. Where treatments were found to be significantly different, pairwise multiple comparison procedures (Tukey Test) were carried out between the high beta-glucan and each of the low beta-glucan phases and there was a highly significant difference between treatments for plasma cholesterol (P = 0.009) and for LDL-cholesterol concentrations (P < 0.001). The differences in plasma cholesterol (6.42 +/- 0.7, 6.14 +/- 0.53, 6.44 +/- 0.67 mmol/L) and LDL-cholesterol (4.59 +/- 0.59, 4.17 +/0.58, 4.52 +/- 0.65 mmol/L) between high beta-glucan and each of the low beta-glucan treatments were significant (P < 0.05). The effect on LDLs (9% lower) is among the highest reported. The results of this study confirm that beneficial reductions in plasma cholesterol and LDL-cholesterol concentrations can be obtained with beta-glucan incorporated into a variety of foods.

Tailoring enzymes for biofuel production with reference to Australian biomass

Manufacture of biofuels from existing biomass may provide a sustainable alternative to the extensive utilization of fossil fuels. Biomass offers environmental advantage over fossil fuels as it is a renewable energy source with low sulphur and nitrogen content and is carbon neutral over its production and utilization. Ranges of biomass are reported worldwide to be suitable raw material for bioethanol production. These can be generally classified into three groups; sucrose based (sugar cane), starch based (corn, wheat and barley) and lignocellulosic (which is mostly comprised of lignin, cellulose and hemicelluloses in grasses, wood and straw) materials. However, the limited supply of two biomass groups (sucrose and starch) will not satisfy society’s growing energy demands; thus biofuel technology based on lignocelluloses is under intense investigation. The main bottleneck in lignocellulosic biomass conversion for biofuel production is the enzymatic depolymerisation of cell wall polysaccharides into fermentable sugars. Protein engineering has recently been used to improve the performance of lignocelluloses degrading enzymes, as well as proteins involved in biofuel synthesis pathways. We have produced a recombinant enzyme that has the ability to produce monomeric sugars from a complex substrate. This presentation will summarize current efforts to develop an enzymatic treatment which would facilitate the economical processing of biomass available in Australia for bioenergy generation.

Fermentative production of rhamnosidase from Staphylococcus xylosus MAK 2 in a bioreactor for bio-energy generation

Increasing concern about the environment, food and feed shortages and hike in the price of petroleum have stimulated interest in new ways of producing biofuels. The interest is rapidly increasing towards converting agricultural wastes to commercially valuable products. Biofuels made from waste biomass can offer immediate and sustained greenhouse gas advantages. In this direction, we are focusing on Citrus processing waste, a byproduct of juice manufacture, which contains high amount of flavonoids and polysaccharides. There is a considerable industrial interest in the enzymatic transformation of flavonoids to hydrolysis products; that offers a pathway to bio-energy generation. Rhamnosidase of bacterial origin are very few and thus are potentially subject for research.

Staphylococcus xylosus, Gram positive cocci, a nonpathogenic member of CNS family, isolated from soil was used to produce α-L-rhamnosidase. This new strain, so far unknown for the production of α-L-Rhamnosidase, was identified and characterized as Staphyloccocus sp. through biochemical tests and 16S DNA sequence analysis. Effect of various medium and process parameters like pH, temperature, aeration and agitation rates and inducer concentration were studied. Further, the enzyme activity was enhanced by adding the inducer and divalent metal ion to the optimised fermentation medium. We have recovered important sugars “rhamnose” and “galacturonic acid” from the processed waste which would be utilized for ethanol production. This presentation will summarize current efforts to develop an enzymatic treatment which would facilitate the economical processing of citrus waste for bioenergy generation.

Updates on naringinase : structural and biotechnological aspects

Naringinases has attracted a great deal of attention in recent years due to its hydrolytic activities which include the production of rhamnose, and prunin and debittering of citrus fruit juices. While this enzyme is widely distributed in fungi, its production from bacterial sources is less commonly known. Fungal naringinase are very important as they are used industrially in large amounts and have been extensively studied during the past decade. In this article, production of bacterial naringinase and potential biotechnological applications are discussed. Bacterial rhamnosidases are exotype enzymes that hydrolyse terminal non-reducing α-l-rhamnosyl groups from α-l-rhamnose containing polysaccharides and glycosides. Structurally, they are classified into family 78 of glycoside hydrolases and characterized by the presence of Asp567 and Glu841 in their active site. Optimization of fermentation conditions and enzyme engineering will allow the development of improved rhamnosidases for advancing suggested industrial applications.

Arthroscopic repair of humeral avulsion of glenohumeral ligament lesion : anterior and posterior techniques

Humeral avulsion of the inferior glenohumeral ligament complex is an unusual pathology, often implicated in traumatic shoulder instability. Traditional open techniques involve at least partial detachment of the subscapularis, and arthroscopic techniques are limited by neurovascular boundaries. The technique described here presents an anterior and posterior arthroscopic approach that can be used individually or in combination to treat different types of humeral avulsion of the inferior glenohumeral ligament lesions. The anterior approach is based on anatomic guidelines described in the literature. The posterior approach is based on the arthroscopic and cadaveric anatomic studies described by one of the authors (D.N.B.); use of the recently described axillary pouch portal (Bhatia portal) permits safe arthroscopic instrumentation access in the entire inferior glenohumeral recess and provides adequate access to the posteroinferior aspect of the humeral head. The differential mattress stitch technique ensures secure fixation of the avulsed ligaments and eliminates excessive capsular redundancy. Technical tips to avoid complications are discussed, and a detailed rehabilitation protocol is presented.