In order to explore the influence of reaction temperature on the product composition, the effect of continuous temperature change (22 degrees C-60 degrees C, +/-0.1 degree C) on hydrolysis of yeast beta-glucan by endo-beta-1,3-glucanase was determined by using self-developed Biochem-temperature Characteristic Apparatus. The activation energy of enzymatic hydrolysis of yeast beta-glucan was 84.17 kJ/mol. The optimum temperature represented by accumulation of products decreased exponentially within a certain period of time. The components of the products were changed with reaction temperature. The length of oligosaccharides decreased with the increase of temperature. The main products were laminaribiose and laminaritriose at the temperature higher than 46 degrees C, while the main products were laminaripentaose and larger molecular weight components at the temperature lower than 30 degrees C. The results can provide precise parameters to control the reaction temperature of the production of 1,3-beta-D-glucooligosaccharides.
Enzyme Activation ; Glucan Endo-1,3-beta-D-Glucosidase ; chemistry ; metabolism ; Hydrolysis ; Oligosaccharides ; chemistry ; metabolism ; Temperature ; Yeasts ; metabolism ; beta-Glucans ; metabolism

OBJECTIVETo develop a spectral assay for determination of pachyman sulfate (PS) in rat plasma and to study the pharmacokinetics after intraperitoneal and intravenous administrations of PS.

METHODThe spectral probe azur A (AA) was used to measure the concentration of PS in rat plasma, since AA could combine the sulfate groups in PS molecules and consequently induced the color change in solution. The optimal wavelengths, concentrations of plasma and AA in reaction system were determined by spectral scanning and serial tests. The plasma PS concentrations were measured at different time after intraperitoneal and intravenous administrations at the dosage of 60 and 20 mg x kg(-1), respectively.

RESULTThe optimal detecting wavelength was 620 nm. The maximum concentration of plasma and the optimal concentration of AA were 1.25% and 8.24 x 10(-5) mol x L(-1) in reaction system, respectively. The calibration curve was linear over the range of 0-10 mg x L(-1) with a correlation coefficiency of 0.995 9. The mean recovery was 100. 55%. The relative standard deviation (RSD) of intra-group and inter-group were all less than 5%. After intraperitoneal and intravenous administrations, the corresponding elimination half-lives were 319.09 min and 204.85 min, respectively. The elimination of PS in blood matched the open model of one compartment and first-order elimination. The bioavailability of PS via intraperitoneal injection was 69.12%.

CONCLUSIONThe spectral probe AA was convenience, sensitive, accurate and steady to use for measuring the concentration of PS in the blood of rats; this made the research work of PS-pharmacokinetics easy and concise.

Animals ; Glucans ; administration & dosage ; blood ; pharmacokinetics ; Infusions, Intravenous ; Injections, Intraperitoneal ; Male ; Poria ; chemistry ; Rats ; Rats, Sprague-Dawley ; Spectrophotometry ; methods

SHG was sulfated by chlorosulfonic acid-pyridine method, and six samples which we got were prepared in different reaction conditions. There is a characteristic absorption peak near 260 nm in UV spectra and there are two characteristic absorption peaks near 1240 cm(-1) and 810 cm(-1) in the FT-IR. Degree of sulfation (DS) was calculated by elemental analysis and turbidimetry. Under the same conditions the absorption peaks become strong with the DS increase. The anticoagulant activity of SHG and sulfated modification samples was evaluated by the classic coagulant assays of prothrombin time (PT), activated partial thrombin time (APTT) live enzymes, and plasma thrombin time (TT). Results show that sulfated SHG has a good anticoagulant activity in vitro, and DS increased activity within a certain range.
Animals ; Anticoagulants ; chemistry ; isolation & purification ; pharmacology ; Blood Coagulation Tests ; Fabaceae ; chemistry ; Glucans ; chemistry ; isolation & purification ; pharmacology ; Partial Thromboplastin Time ; Plants, Medicinal ; chemistry ; Prothrombin Time ; Rabbits ; Spectrophotometry, Ultraviolet ; Spectroscopy, Fourier Transform Infrared ; Sulfonic Acids ; chemistry ; Thrombin Time

The invasive fungal infections (IFI) in immunocompromised patients are associated with a high mortality rate and diagnostic difficulty. Serological methods such as aspergillus galactomannan assay (GM test) and (1, 3)-beta-D glucan (BG) assay (G test) can be used as an adjunctive method for IFI diagnosis based on their characteristics of easy-operating, rapidness and high sensitivity. Compared with GM test, G test can be more widely used except for the diagnosis of aspergillosis. The purpose of this study was to investigate the value of G test in the diagnosis of IFI in patients with hematological disorders. The plasma was collected from 162 suspected IFI patients with hematological disorders in Beijing Daopei Hospital, including 85 patients after chemotherapy and 77 patients after stem cell transplantation from May 2007 to May 2008, BG level was measured with MB-80 Microbiology Kinetic Rapid Reader and the measured results together with the clinical characteristics were retrospectively analyzed. According to the European Organization for Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) criteria, there were 2 patients diagnosed as proven IFI, 18 as probable IFI, 75 as possible IFI and 67 as no IFI. The results showed that at a cutoff of 20 pg/ml, the sensitivity and specificity of G test were 75% and 91% respectively, with a positive predictive value (PPV) of 71.4% and a negative predictive value (NPV) of 92.4%. 51 out of the 75 possible IFI patients with elevated BG level were responsive to antifungal treatment but non responsive to broad-spectrum antibiotics, retrospectively were diagnosed as IFI, suggesting that G test improved the IFI diagnostic rate by 31.4%. In conclusion, G test is a rapid and simple method for early diagnosis of IFI in patients with hematological disorders.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Hematologic Diseases ; blood ; diagnosis ; microbiology ; Humans ; Male ; Middle Aged ; Mycoses ; blood ; diagnosis ; Plasma ; chemistry ; Young Adult ; beta-Glucans ; blood

OBJECTIVETo study the effect of laminarin polysaccharide (LP) on the activity of matrix metalloproteinase of photoaging skins.

METHODKunming SPF mice were prepared with back hair shaved, and randomly divided into the control group, the model group, the LP low does group (LP-L, 1 mg x kg(-1)), the LP high dose group (LP-H, 5 mg x kg(-1)) and the Vit E (100 mg x kg(-1)) group. They were abdominally injected with drugs twice on a daily basis. Except for the control group, all groups were exposed to ultraviolet rays for 1 hour every day, five times on a weekly basis, with accumulated exposure dose of UVB being 21.60 J x cm(-2) and accumulated exposure dose of UVA being 84.02 J x cm(-2). Eight weeks later, exposed back skins were collected to detect thickness of dermis by HE stain, content of hydroxyproline (Hyp) by chemical colorimetry, and serum MMP-1 and TIMP-1 content by ELISA. In addition, matrix metalloproteinase-1 (MMP-1) mRNA and relative content of tissue inhibitor of metalloproteinase-1 (TIMP1) mRNA was analyzed with Real-time PCR.

RESULTCompared with the model group, the LP-H group could significantly increase the thickness of dermis, skin Hyp content and serum TIMP-1 level, and decrease relative content of MMP-1 mRNA in skin and MMP-1 content in serum.

CONCLUSIONLP can regulate the metabolism of collagen photoaging skins by adjusting the activity of matrix metalloproteinase.

Animals ; Female ; Glucans ; Matrix Metalloproteinase 13 ; biosynthesis ; genetics ; metabolism ; Mice ; Plant Extracts ; chemistry ; pharmacology ; Plants, Medicinal ; chemistry ; Polysaccharides ; chemistry ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Skin Aging ; drug effects ; physiology ; radiation effects ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; metabolism ; Ultraviolet Rays

OBJECTIVETo study anti-HIV activity and mechanism of Cynanchum otophyllum glucan sulfate in vitro.

METHODAnti-HIV-1 activity was detected with syncytial formation assay and quantitative P24 enzyme-linked immunosorbent assay (ELISA); cytotoxicity was tested with MTT colorimetric assay. Antiviral mechanism was investigated by fusion inhibition, time of addition and pre-treatment experiments.

RESULTThe 50% inhibition concentrations (IC50) of PS20 for HIV-1(IIIB), HIV-1(Ada-M), and HIV-1(Bal), were 0.26, 0.46, 0.90 micromol x L(-1), respectively. Studies on antiviral mechanism of PS20 showed that target molecule may be viral envelope protein.

CONCLUSIONThe results suggested that PS20 had high anti-HIV activity and was worth to be studied further.

Anti-HIV Agents ; pharmacology ; Cell Fusion ; Cell Line ; Cell Proliferation ; drug effects ; Cynanchum ; chemistry ; Glucans ; isolation & purification ; pharmacology ; HIV-1 ; drug effects ; Humans ; Inhibitory Concentration 50 ; Plant Extracts ; pharmacology ; Plant Roots ; chemistry ; metabolism ; Viral Envelope Proteins ; drug effects

An extracellular lipase from Aureobasidium pullulans was obtained and purified with a specific activity of 17.7 U/mg of protein using ultrafiltration and a DEAE-Sepharose Fast Flow column. Characterization of the lipase indicated that it is a novel finding from the species A. pullulans. The molecular weight of the lipase was 39.5 kDa, determined by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme exhibited its optimum activity at 40 °C and pH of 7. It also showed a remarkable stability in some organic solutions (30%, v/v) including n-propanol, isopropanol, dimethyl sulfoxide (DMSO), and hexane. The catalytic activity of the lipase was enhanced by Ca²⁺ and was slightly inhibited by Mn²⁺ and Zn²⁺ at a concentration of 10 mmol/L. The lipase was activated by the anionic surfactant SDS and the non-ionic surfactants Tween 20, Tween 80, and Triton X-100, but it was drastically inhibited by the cationic surfactant cetyl trimethyl ammonium bromide (CTAB). Furthermore, the lipase was able to hydrolyze a wide variety of edible oils, such as peanut oil, corn oil, sunflower seed oil, sesame oil, and olive oil. Our study indicated that the lipase we obtained is a potential biocatalyst for industrial use.
Ascomycota/enzymology* ; Calcium ; Catalysis ; Corn Oil/metabolism* ; Detergents/chemistry* ; Enzyme Stability ; Fungal Proteins/chemistry* ; Glucans/chemistry* ; Hexanes/chemistry* ; Hydrogen-Ion Concentration ; Hydrolysis ; Industrial Microbiology ; Lipase/chemistry* ; Manganese/chemistry* ; Olive Oil/metabolism* ; Peanut Oil/metabolism* ; Sesame Oil/metabolism* ; Substrate Specificity ; Sunflower Oil/metabolism* ; Surface-Active Agents ; Temperature ; Zinc/chemistry*

OBJECTIVETo investigate regulation function of anodonta glucan HBP-A on chondrocytes through Wnt pathway in vitro.

METHODSRat chondrocytes were cultured and differentiated induced with IL-1beta (10 ng/ml) in vitro. Chondrocytes were divided into five groups:IL-13 group,IL-1beta + IWP-2 (5 microM,Wnt pathway inhibitor) group, IL-1beta + HBP-A (0.3 mg/ml) group and IL-1beta + IWP-2 + HBP-A group. Wnt-3a, beta-catenin (24 h,48 h,72 h) and MMP-13(72 h) genes expression were detected by Rt-PCR, while beta-catenin, MMP-13, Sox-9 and coll-II (48 h) protein expression were measured by Western-blot.

RESULTSAfter induction of IL-1beta, gene expression of Wnt-3a, beta-catenin and MMP-13 were increased,so were the protein expression of beta-catenin and MMP-13. In contrast,protein expression of Sox-9 and Coll-II were declined. Following addition of HBP-A, Wnt-3a, beta-catenin and MMP-13 were shown as induction of IL-1beta, but protein expression of Sox-9 and Coll-II were upgraded. Combining HBP-A with IWP-2 led to the lowest level in Wnt-3a, beta-catenin gene and beta-catenin protein expression and highest expression of Sox-9 protein.

CONCLUSIONHBP-A could not only delay the differentiation of chondrocytes through downgrading the signal expression of Wnt/beta-catenin,but also adjust the expression of Wnt-3a, beta-catenin and Sox-9 when combinated with the Wnt inhibitor.

Animals ; Anodonta ; chemistry ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chondrocytes ; cytology ; drug effects ; metabolism ; Glucans ; pharmacology ; Interleukin-1beta ; metabolism ; Rats ; Wnt Signaling Pathway ; drug effects ; Wnt3A Protein ; genetics ; metabolism ; beta Catenin ; metabolism

The beta-glucans derived from yeast cell walls have been reported for having many immunomodulatory activities in vivo and in vitro. In this study, Aureobasidium-derived soluble branched (1,3-1,6) beta-glucan (Sophy beta-glucan) was checked for natural killer (NK) activity and for the production of IFN-gamma and IL-4 in Leishmania amazonensis infection. The main experiment was performed with a group of female C57BL/6 and BALB/c mice, orally supplemented with 5% of Sophy beta-glucan and infected with promastogotes of L. amazonensis (1 x 10(7)) into the footpad. Increase in the footpad thickness with time was observed in BALB/c mice in spite of the oral Sophy beta-glucan supplement, but it was less in C57BL/6 mice. The difference in overall mean footpad thickness between 'infection only' versus 'infection + glucan' groups was statistically significant (P < 0.001). High NK activity in C57BL/6 than BALB/c mice was observed in 'glucan only' group compared to the control group and also in 'infection + glucan' group compared to 'infection only' group. The difference in the NK activity among these groups was significant (P < 0.05). The IFN-gamma level increased at weeks 7 and 8 post-infection in C57BL/6 mice and was significantly high in 'infection + glucan' group compared to the 'infection only' group (P < 0.05). IL-4 levels did not increase up to detectable levels throughout the study. The results led a conclusion that Sophy beta-glucan enhances NK activity and cellular immunity in L. amazonensis-infected mice.
Administration, Oral ; Animals ; Ascomycota/*chemistry ; Cytotoxicity Tests, Immunologic ; Female ; Foot/pathology ; Glucans/administration & dosage/*isolation & purification/pharmacology/*therapeutic use ; Immunologic Factors/administration & dosage/*isolation & purification/pharmacology/*therapeutic use ; Interferon-gamma/biosynthesis ; Interleukin-4/biosynthesis ; Killer Cells, Natural/drug effects/*immunology ; Leishmania mexicana/*immunology ; Leishmaniasis, Cutaneous/*drug therapy/immunology/pathology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Severity of Illness Index ; Time Factors

We investigated the immunostimulatory effects of a novel beta-glucan purified from Paenibacillus (P.) polymyxa JB115 on bone marrow-derived dendritic cells (DCs), a type of potent antigen-presenting cells. beta-glucan isolated from P. polymyxa JB115 enhanced the viability and induced the maturation of DCs. beta-glucan markedly increased the cytokine production of DCs and surface expression of DC markers. In addition, DCs treated with beta-glucan showed a higher capacity to stimulate allogeneic spleen cell proliferation compared to those treated with medium alone. These results demonstrate the effect of beta-glucan on DC maturation and may increase the use of beta-glucan.
Animals ; Bone Marrow Cells/cytology/*drug effects/*immunology ; Cell Survival/drug effects/*immunology ; Dendritic Cells/cytology/*drug effects/*immunology ; Flow Cytometry ; Immunophenotyping/methods ; Interleukin-12/analysis/immunology ; Mice ; Mice, Inbred BALB C ; Nitric Oxide/analysis/immunology ; Paenibacillus/*chemistry ; Tumor Necrosis Factor-alpha/analysis/immunology ; beta-Glucans/isolation & purification/*pharmacology