New possibilities in treatment of chronic urinary tract infections-vaccines, autovaccines and beta glucans

Background:Repeated infections of urinary tract manifest an increasing trend and became the most common infection in developed world.In addition,currently common increase in resistant microbes including uropathogens underlines the need for new ways of urinary tract infection(UTI)treatment.One of the possibilities is vaccination.However,commercially available bacterial vaccines are not always reliable,leading to the proposition of using autovaccines prepared from uropathogens of individual patients.It is a well-known fact that UTIs are often accompanied by depressed immunity.Methods:To respond to both problems,we tried to use yeast-derived,insoluble beta glucans for restoration of depressed immune system and acceleration of the effects of autovaccines.In all patients we repeatedly tested bacterial load in urine.In addition,from the same samples we measured the levels of inflammatory proteins,albumin,orosomukoid,IgA,C-reactive protein.Complex evaluation of inflammatory response and cellular immunity in blood was performed,too.Results:Immediately after application of the first dose we observed significant improvements of clinical conditions which were persistent throughout the entire study.Conclusions:Beta glucan is optimal addition to autovaccines,as it is natural,active,safe and inexpensive.We propose that the possibility to use beta glucans in application of autovaccines is the first step in preparation of qualitative new type of autovaccines against UTI.

Sensorial Evaluation and Physical Chemical Characterization of a Beverage Based on Whey and β-Glucans as a Potential Prevention of Nonalcoholic Steatohepatitis

Whey and B-glucans are functional food ingredients that contribute to the prevention of non-communicable diseases. The objective of the study was to develop a functional beverage based on whey, B-glucans, and blackberry concentrate for the prevention of nonalcoholic steatohepatitis, determining its sensory, physicochemical, and nutritional qualities. The following phases were developed: 1) formulation with 75%, 85%, and 90% whey and 25%, 15%, and 10% blackberry concentrate, with a fixed amount of 0.243 g β-glucans in 100 ml;2) microbiological analysis to ensure the safety of the beverage;3) sensory analysis with untrained panelists to assess the degree of preference (appearance, color, odor, flavor, viscosity, acidity, sweetness), and general acceptance, with a seven-point hedonic scale to rate the level of acceptability;4) physicochemical and nutritional characterization of the beverage with the highest acceptance. An experimental design using complete random blocks was employed. Data analysis was performed with variance (ANDEVA) and mean DUNCAN separation at a 95% confidence level, using SAS®. A correlation study was conducted on acceptance analysis. A nutritional label was developed for the control beverage and the treatment with the highest acceptance. The formulation with 75% whey, 25% blackberry concentrate, and 0.243 g of β-glucan obtained the highest acceptance, with flavor and sweetness being the attributes that contributed to its acceptance. Nutritionally, the beverage with 80 cal, would be considered fortified with fiber (2 g), free of cholesterol, sodium, and saturated fat, according to the Central American Technical Regulation on Nutritional Labeling (Version 67.01.60:10). It is recommended that the effect of this beverage be measured to determine the benefits.

黑曲霉DL08内切葡聚糖酶基因的克隆表达及酶学性质研究

通过反转录PCR(RT-PCR)从黑曲霉(Aspergillus niger)DL08中提取内切葡聚糖酶基因(GeneBank No.KJ437592),PCR测序表明该基因全长999个核苷酸,编码332个氨基酸,预测相对分子质量为36.75 kDa,等电点(pI)为4.38,命名eg1。结构域分析表明,该蛋白包括18个氨基酸构成的信号肽和C末端1个糖基水解酶家族5的催化结构域。重组内切葡聚糖酶蛋白通过Ni-NTA亲和层析柱纯化,酶学性质研究表明,以羧甲基纤维素钠为底物时重组酶最适作用pH为5.0,最适作用温度为45℃。通过薄层层析法检测重组内切葡聚糖酶酶解1%羧甲基纤维素钠的产物,主要为连续寡糖。这些特性为纤维素酶酶解纤维素生产生物化学品和可再生生物燃料提供技术参考。

Glucan对氰化钾致肝细胞损伤的保护作用

目的观察氰化钾对肝细胞的损伤作用和Glucan的干预效果及作用机制。方法体外培养的SD乳鼠肝细胞 ,用氰化钾诱导损伤 ,用Glucan预先处理损伤组 ,观察正常对照组、损伤组、Glucan保护组肝细胞的活性 ,并检测培养上清液中的乳酸脱氢酶、超氧化物歧化酶活性和一氧化氮含量的变化。结果氰化钾可使肝细胞中的乳酸脱氢酶漏出增多 ,同时培养液中超氧化物歧化酶活性降低 ,一氧化氮含量减少 ,预先应用Glucan后 ,可部分逆转这些改变。

Row Spacing Affects Biomass Yield and Composition of Kenaf (<i>Hibiscus cannabinus</i>L.) as a Lignocellulosic Feedstock for Bioenergy

Kenaf (Hibiscus cannabinus L.) is a warm-season annual. Kenaf fibers are commonly used for paper pulp and cordage, but it is also a promising lignocellulosic feedstock for bioenergy production, although optimum plant density for biomass production has not been determined for the northern region of the USA. The objective of this study was to determine the best plant density and row spacing of kenaf to maximize biomass yield and chemical composition for biofuel conversion. The experiments were conducted at Fargo and Prosper, ND, in 2010 and 2011. The experiment was a randomized complete block design with a split-plot arrangement where the main plot was tworowspacings (30 and60 cm) and the sub-plot fourplant densities (32, 16, 8, and 4 plants·m-2). Row spacing had a significant effect on both biomass and biofuel yield. Narrower rows had higher biomass and biofuel yield. Maximum biomass and estimated biofuel yield was obtained with the two highest plant densities of 16 and 32 plants·m-2 and fluctuated between 9.45 and 10.22 Mg·ha-1 and 1354 and1464 L·ha-1, respectively. Stem diameter increased with a decrease in plant density. Chemical composition varied with plant density;glucan (27%) and xylan (9.8%) content were lower at the lowest plant density. Ash content was not different among plant densities but it is interesting to mention the very low ash content of kenaf (0.15%). According to the results of this study, it is recommended to plant kenaf at 30-cm rows with a plant density of 16 to 32 plants·m-2 to maximize biomass yield. Kenaf has a tremendous potential as a cellulosic feedstock for biofuel and green chemicals in the Northern Great Plains because of high biomass yield and low ash content.

Glucan在Con A所致免疫性肝损伤中的作用

目的观察Glucan对刀豆蛋白A(ConA)所致小鼠免疫性肝损伤的干预效果及其作用机制。方法ConA尾静脉注射制备小鼠免疫性肝损伤模型,在ConA注射前1h腹腔注射Glucan,观察正常对照组、ConA损伤组、Glucan/ConA组肝损伤的情况,检测血清谷丙转氨酶(ALT)、α肿瘤坏死因子(TNF-α)及组织中一氧化氮(NO)含量的变化,并用DNAladder检测肝细胞凋亡的变化。结果ConA尾静脉注射后,出现明显的肝损伤,血清ALT、TNF-α及组织中NO水平明显升高,DNAladder检测可见明显的梯状条带;预先应用Glucan后,可减轻这些改变。结论Glu-can对ConA所致小鼠免疫性肝损伤有保护作用。

脱乙酰几丁质对烟草叶片一些生理生化特性的影响

以温室栽培生长7～8周的烟草（Nicotianatobacum）第4片真叶为材料，测定了脱乙酰几丁质对细胞膜透性的影响。结果发现，脱乙酰几丁质处理的烟草叶片的电解质、可溶性蛋白质的外渗量和胞外可溶性过氧化物酶的活性均比对照组大，并且随所用的脱乙酰几丁质浓度增加而增大。Mg2＋、Ca2＋、Al3＋能抑制脱乙酰几丁质诱导的蛋白质外渗。脱乙酰几丁质（500μgml-1）处理的烟草叶片的几丁醇和β－1，3－葡聚精酶的活力，在处理6h后开始上升，至36h活力达最高峰，分别是对照组的1．88倍和1．51倍。5mmolL-1Ca2＋对这种诱导具有抑制作用。伴随着对几丁酶和β-1，3-葡聚糖酶的诱导，处理组的可溶性蛋白含量也在处理后36h达到最高峰。

漂白针叶木浆酶促打浆效果的研究

研究了纤维素酶和β-葡聚糖酶的预处理对节约打浆能耗的影响,通过测定纸浆的黏度、保水值、还原糖含量以及纸张的强度性能来反映酶处理纸浆的效果。结果表明,酶处理的确可以有助于节约打浆能耗,提高纸浆的滤水性能,增加纤维的保水值。同时,纸张的强度性能也得到改善。

PREPARATION AND STRUCTURE OF FIVE DERIVATIVES OF β-(1→3)-D-GLUCAN ISOLATED FROM PORIA COCOS SCLEROTIUM

A new solvent of cellulose (1.5 mol/L NaOH/0.5 mol/L urea aqueous solution) was used as one of the homogeneous reaction media of polysaccharides for methylation, hydroxyethylation and hydroxypropylation. A water insoluble β -(1—>3)-D-glucan, sample PCS3- isolated from fresh sclerotium of Poria cocos was sulfated in dimethyl sulfoxide (Me 2 SO), carboxymethylated in NaOH, isopropanol solution, as well as methylated, hydroxyethylated and hydroxypropylated in the new solvent system, respectively, to obtain five water-soluble derivatives coded as S-PCS3- C- PCS3- M-PCS3- HE-PCS3- and HP-PCS3- Their chemical structure and distribution of substitution were characterized by infrared spectroscopy (IR), elementary analysis (EA), 1 H-NMR, 13 C-NMR, 2D-COSY, 2D-TOCSY and 2D- 1 H-detected 1H 13C HMQC spectra. The results reveal that the relative reactivity of hydroxyl groups of the β -(1-?3)-D-glucan is in the order C-6 > C-4 > C-2 on the whole. The substitution of the samples S-PCS3- C-PCS3- and M-PCS3- occurred mainly at C-6 position and secondly at C-4 and C-2 positions, and that of HE-PCS3- occurred at C-6 and C-4 positions and of HP-PCS3- almost completely occurred at C-6 position. The degrees of substitution (DS) obtained from 13 C-NMR range from 0.23 to 1.27. The water solubility of the derivatives is in the order S-PCS3- >C-PCS3- >M-PCS3- >HE-PCS3- >HP-PCS3- This work provides a novel and nonpolluting process for the methylation, hydroxyethylation and hydroxypropylation of β -(1—>3)-D-glucan.

Glucan-immunostimulant,adjuvant,potential drug

β-glucans belong to a group of biologically active natural compounds called biological response modifiers.These substances represent highly conserved structural components of cell walls in fungi,yeast,grain and seaweed.Despite almost 160 years of intensive research,the exact mechanisms of their action remain unsolved.The significant role of glucans in cancer treatment,infection immunity,stress reduction and restoration of damaged bone marrow has already been established.The present review focuses on the various less known but potentially significant roles glucans might play in medicine.In summary,glucan might represent the most important natural immunomodulator.

Structural Characterization of Glucan Produced by Lactobacillus sake L-7 from Sausage

A linear glucan was produced by Lactobacillus sake L-7 isolated from homemade sausage. Cultivation of the strain in Man–Rogosa–Sharpe(MRS) medium containing 50 g/L sucrose yielded 5.3 g/L of purified exopolysaccharide(EPS). The EPS was characterized by gas chromatography(GC), Fourier-transform infrared(FT-IR) spectroscopy, high-performance sizeexclusion chromatography(HPSEC), nuclear magnetic resonance(NMR) spectroscopy, and scanning electron microscopy(SEM). The monosaccharide composition of the EPS was glucose, and its molecular weight was 1 × 10~7 Da. The FT-IR and NMR spectra revealed that the L-7 EPS was a linear glucan with α-(1 → 6) glucosidic bonds. SEM images of the dried EPS revealed a hollow tubular structure. The water solubility index and water holding capacity of L-7 EPS were 96 and 272%, respectively. The results of hydrolysis indicated that L-7 EPS was not susceptible to hydrolysis by physiological barriers and can be used as a soluble dietary fiber with health benefits. All these characteristics suggest that L-7 EPS might have potential applications in the food, cosmetic, and pharmaceutical industries.

Characterization and Anti-tumor Activity of Glycopeptides from Ganoderma sinensis

The water-soluble part（GS） of Ganoderma sinense Zhao, Xu et Zhang was divided into high molecular（GS-H） and low molecular（GS-L） parts by Cellulose Super Filtration, and GS was also fractionated into four fractions, GS-1, 2, 3, and 4 by ethanol precipitation according to their molecular weights. Chemical analysis shows that GS and GS-1, 2, 3, 4 were complexes of polysaccharide and peptide. The fractions with molecular weights over 4000, GS-1, 2, 3, and GS-H show anti-tumor activities, however, the fractions with molecular weights lower than 4000, GS-4, and GS-L have no anti-tumor activity, indicating that the anti-tumor activity of Ganoderma Sinensis was caused by glucopeptides with molecular weight ranging from 4000 to 20000. Two purified glucopeptides, GS-6b and GS-7b were obtained from GS-H by ion-exchange and gel-permeation chromatography. Their molecular weights, glycosidic linkages, and configurations were detected by means of IR spectrum, sugar composition analysis, and methylation analysis. The polysaccharide parts of GS-6b and GS-7b had glucan backbone consisting of β-1→3 Glc, and side chain containing glucosyl, mannosyl, fucosyl, xylosyl, galactosyl, and glucuronic acid residues attached on 1-2, 1-4, 1-6 positions of the backbone of GS-6b, or 1-6, 1-4 positions of the backbone of GS-7b. The peptide parts in GS-6b and GS-7b were composed of 10 kinds of amino acids, including Asp, Ser, Arg, Gly, Thr, Pro, Ala, Val, Met, and Lys.

Transient expression of the <i>Arabidopsis thaliana</i>callose synthase PMR4 increases penetration resistance to powdery mildew in barley

Localized cell wall thickenings, so called papillae, are a common plant defense response to fungal attack at sites of penetration of the plant cell. The major constituent of papillae is callose, a (1,3)-β-glucan polymer, which contributes to slowing or blocking the invading fungal hyphae. In the model plant Arabidopsis thaliana, we could recently show that the overexpression of PMR4(POWDERY MILDEW RESITANT 4), which encodes a stress induced callose synthase, results in complete powdery mildew resistance. To evaluate if these findings are also transferable to monocot crops, we transiently expressed PMR4 under control of the 35S promoter in leaves of barley (Hordeum vulgare) seedlings, which were subsequently inoculated with the virulent powdery mildew Blumeria graminis f. sp. hordei. Fusion of the green fluorescent protein (GFP) to PMR4 allowed the identification of successfully transformed barley cells, which showed an increased penetration resistance to B. graminis compared to control cells that express only GFP.PMR4-GFP localized in a similar pattern at the site of attempted fungal penetration as observed inA. thaliana, which suggests that similar transport mechanisms of the callose synthase might exist in dicot and monocot plants.

Administration of Yeast Glucan on Immunity of Offspring in Turbot Scophthalmus maximus: A Trans-Generational Immune-Enhancing Effect

β-glucan has been shown to increase immunity and survival in various juvenile and adult fish both in freshwater and marine aquaculture species as well as in marine fish larvae. However, information about the trans-generational immune-enhancing effects of β-glucan remains rather limited. Here we clearly show that dietary intake of β-glucan enhanced the levels of C3, Bf and lysozyme in the serum of turbot as well as in their eggs released. We also show that yeast glucan induced a significant increase in lysozyme activity in both the serum and eggs. Moreover, the embryos derived from yeast glucan-treated turbot were more resistant to bacterial challenge than control embryos. By contrast, the administration of yeast glucan on female turbots had little influence on the egg development and embryonic development. Collectively, these data indicate that yeast glucan can be safely used to promote the non-specific trans-generational immunity in offspring of turbot.

Analysis of Clinical Characteristics and Diagnostic Value of Fungal Serology in Patients with Invasive Candidiasis

Objective: To evaluate the diagnostic value of (1 - 3)-β-D glucan and mannan assay for invasive candidiasis. Methods: A retrospective study was conducted on 32 cases in the disease group (18 proven invasive candidiasis and 14 probable invasive candidiasis) and 48 cases in the control group. The subjects were recruited from January 2018 to March 2019 in Clinical Laboratory of Hainan General Hospital. All subjects were detected by (1 - 3)-β-D glucan and mannan assay. Results: The mean concentration of (1 - 3)-β-D glucan in the disease group was 97.45 (43.23, 224.35) pg/ml and it was significantly higher than the mean concentration of the control group which was 49.85(41.91, 56.07) pg/ml (P = 0.005). The mean concentration of mannan in the disease group and the control group were 161.36 (34.96, 224.49) pg/ml and 25.80 (25.00, 29.31) pg/ml, respectively, which were significantly different (P < 0.001). The sensitivity, specificity, positive predictive value and negative predictive value of (1 - 3)-β-D glucan assay were 59.38%, 89.58%, 79.17%, 76.79%, respectively. The sensitivity, specificity, positive predictive value and negative predictive value of mannan assay were 65.63%, 95.83%, 91.30%, 80.70%, respectively. The sensitivity, specificity, positive predictive value and negative predictive value of combination of two types of assays were 81.25%, 85.42%, 78.79% and 87.23%, respectively. Conclusions: Combination of (1 - 3)-β-D glucan and mannan assay can improve diagnostic specificity and it has essential clinical diagnostic value for invasive candidiasis.

Identification of genes involved in the formation of soluble dietary fiber in winter rye grain and their expression in cultivars with different viscosities of wholemeal water extract

The grain of rye(Secale cereale L.) used for baking contains a large amount of non-starch polysaccharides,making it an excellent component of functional foods. But rye grain intended for alcohol production and forage use should have a reduced content of these polysaccharides. A comprehensive parameter that can predict the best field of application for winter rye grain is the viscosity of its wholemeal water extract.However, our understanding of the genetic background underlying this key trait and associated features of rye grain is poor. By analyzing six Russian winter rye cultivars, we identified the most contrasting forms and characterized the peculiarities of their water-soluble carbohydrates capable of influencing the viscosity of water extracts. Then, using phylogenetic and transcriptomic analyses, we identified in the rye genome many genes encoding putative glycosyltransferases and glycosylhydrolases responsible for the synthesis and degradation of arabinoxylans, mixed-linkage glucans, cellulose, and some other polysaccharides. We determined the dynamics of m RNA abundance for these genes at three stages of kernel development. Comparisons of gene expression levels in two contrasting cultivars revealed specific members of multigene families that may serve as promising targets for manipulating non-starch polysaccharide content in rye grain. High-viscosity cultivars were characterized by up-regulation of many glycosyltransferases involved in the biosynthesis of arabinoxylans and other cell-wall polysaccharides,whereas low-viscosity cultivars showed up-regulation of several genes encoding polysaccharidedegrading enzymes.

ISOLATION, PROPERTIES AND CHEMICAL STRUCTURE OF A GLUCAN FROM GLYCYRRHIZIA URALENSIS FISCH

A new polysaccharide,GBW was obtained homogeneous as determined by HPLC from the roots of Glycyrrhizia uralensis Fisch. The molecular weight was estimated to be 4000. The component sugar was identified as glucose only. The specific rotation [α]^(28)_D (C=0.1,H_2O) was +1200. The α-configuration was established by IR and ^(13)C NMR. Methylation analysis, periodate oxidation, Smith degradation, partial hydrolysis, KI-I_2 reaction and ^(13)C NMR showed that GBW is a new (1→4)linked linear α-D-glucan. The structure is shown as 1.

Direct comparison of immunological effects of various nutritional supplements

Objective:Immunonutrition represents a diet based on the knowledge of basic principles of the immune system and its functions.Despite numerous claims,the direct comparison of the immunostimulating effects of natural modulators is limited,making any conclusions difficult.Our study focused on most common vitamins and immunonutrients and directly compared their effects on various branches of the immune system.Methods:In this study we used a mouse model to evaluate various aspects of immune reactions,namely phagocytic activity,IL-2 secretion,NK cell activity,antibody formation and growth of breast cancer in order to find out the possible effects of six different immunonutrients and their combinations.Results:We found that glucan was in all tests the most active immunomodulator.Synergistic effects were observed only in glucan-selenium and glucan-vitamin C combinations.The rest of immunonutrients had only small activity or no activity at all.Conclusion:Based on our results,we concluded that most of vitamins and minerals have only limited,if any,effects on immune activities including cancer.The current study managed to confirm synergistic effects of the beta glucan-vitamin C and beta-glucan-selenium combinations.More studies on possible positive or negative effects of such combination and on mechanisms of action are important.

Biofilm Formation by <i>Streptococcus mutans</i>and Related Bacteria

Caries is a disease of human dentition characterized by the loss of mineralized surfaces of the tooth;it is an infectious disease of the oral cavity in which biofilms play a causative role. Control of biofilms has traditionally relied on non-specific removal of plaque by mechanical means such as brushing, although it is difficult to remove biofilms by this method. Caries is also a widespread infection in children. Streptococcus mutans and S. sobrinus are important causative agents of caries. They produce a homologous exocellular polysaccharide called glucan, which strongly adheres to the enamel surface. This is a review of oral microbial biofilm formation by S. mutans and other related bacteria.

Preparation of Water Soluble Yeast Glucan by Four Kinds of Solubilizing Processes

(1→3)-β-D-glucan?from the inner cell wall of Saccharomyces cerevisiae is considered a member of a class of drugs known as biological response modifiers (BRM). However the glucan was an insoluble polysaccharide, which could be the major barrier to the utilization of glucan. In this case, the insoluble glucan was convent into a soluble form by four kind of solubilizing processes. The yield, solubility, chemistry structure and immunoprophylaxis efficacy of the soluble products were compared. Our date suggest that urea has a significant effect on yield, and DMSO has a significant effect on solubility. FT-IR spectra, 13C NMR spectra and helix-coil transition analysis demonstrate that the chemistry structure of native and solubilizing glucans have no significant difference. They still have the triple helical structure. The solubility and immunoprophylaxis efficacy assay indicate that the introduction of phosphate group not only enhanced the solubility of glucan, but also improved the survival rate of mice challenged with E. coli.