Combination Of Fructose Research Articles

Interaction of Culture Medium and Artificial Light Type on Pigmentation of Micro-Propagated Opuntia Plants

Opuntia is recognized economically as a significant crop for its nutritional, functional, and therapeutic properties and its potential in the pharmaceutical, cosmetic, and bioenergy industries. Opuntia is conventionally propagated by seed or vegetative propagation via rooted scions or grafting. However, multiplication procedures are insufficient for extensive spreading. One of the alternative techniques is in vitro. For this reason, the objective was to evaluate the growth of different components combined with light conditions to induce callus, embryogenesis, and the possible synthesis of a component in the genus Opuntia. The existence of genes involved in pigment synthesis in genotypes of different fruit colors was complementarily analyzed. In this study, we used different combinations of fructose (2 g/L) with prickly pear juice (2 mL/L), mannitol (4 g/L), silver nitrate (AgNO3: 1 mg/L), 2,4-dichlorophenoxyacetic acid (2,4-D: 2 mg/L), and indole-3-acetic acid (IAA: 1 g/L), with white, blue, and red LEDs (light-emitting diodes) and laser beams. These explants yielded sufficient contents of simple phenols, gallic acid equivalents (GAE: 2283.30 ± 6.29 mg/100 g), and coumaric (2155.0 ± 35.0 mg/100 g) and ferulic (2176 ± 27.9 mg/100 g) acids for the genotype Tapón aguanoso, and chlorogenic acid (CGA: 380.22 ± 22.05 mg/100 g) for the Copena V1 genotype. Upon analyzing the genotypes of different fruit pigmentations, we also observed the following: enzyme 4,5 estradiol dioxygenases can be present in all genotypes (regardless of pigmentation); enzyme 5,3 glucosyltransferases (GTs) could be induced in pigment synthesis in the genotypes of orange to purple fruits. Sequencing primer-amplified fragments for GT showed high similarity to uridine diphospho (UDP)-glucose from other species. This allows us to infer that it is possible to obtain products of high therapeutic value in the near future under controlled conditions.

GLUT5, GLUT7, and GLUT11 expression and Bcl-2/Bax ratio on Breast Cancer Cell Line MCF-7 Treated with Fructose and Glucose.

Fructose and glucose are types of sugars commonly found in the diet that have been linked to cancer development. Glucose transporters (GLUTs) are facilitating the uptake of these hexoses. Expression of GLUT5 is higher in cancer cells than in healthy tissue. GLUT7 and GLUT11 facilitate the transport of glucose and fructose; however, their expression in breast cancer has not been extensively studied. The Bcl-2 family has been known as a regulator of the cell's survival and death. Here, we investigated the effect of the fructose-glucose combination in MCF-7 breast cancer cells on the viability, migration, and expression of GLUT5, GLUT7, GLUT11, and Bcl-2/Bax ratio. Breast cancer cells MCF-7 were treated with fructose, glucose, and combinations of fructose:glucose (75%:25%, 50%:50%, 25%:75%). Cell viability was assessed using an MTT test. Cell migration was examined with a wound-healing assay. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to evaluate the mRNA expression of GLUT5, GLUT7, GLUT11, and Bcl-2/Bax. The viability and migration of MCF-7 breast cancer cells elevated when treated with a combination of fructose and glucose, and glucose alone, compared to fructose alone. The expression levels of GLUT5 and GLUT7 were highest in combination of fructose:glucose (75%:25%). Conversely, the expression of GLUT11 was consistently low across all treated media. The highest Bcl-2/Bax ratio was shown in fructose:glucose combination (25%:75%). The viability, migration, and Bcl-2/Bax ratio are enhanced in the combination media with higher glucose. In contrast, when the fructose composition was higher in the media, expression of GLUT5 and GLUT7 increased.

Trehalose as a Stabilizer of the Lipid Composition of Membranes and the Composition of the Cytosol of Frozen/Thawed Rooster Spermatozoa

Low-temperature semen storage technologies are already being used in poultry conservation programs, but the quality of reproductive material stored in cryobanks varies greatly and cannot always be successfully used for practical purposes. Therefore, it is necessary to improve the compositions of cryoprotective media to improve their quality. This study aimed to investigate the composition of membrane lipids and carbohydrates in the cytosol of rooster spermatozoa, to explain the dose-dependent effect of a combination of trehalose and fructose in cryoprotective media on the preservation of their morphological and kinetic parameters during freezing/thawing, and to determine the most effective diluent composition. Ejaculates were collected from Rhode Island Red roosters (n = 10). The effectiveness of three diluents containing trehalose was evaluated: LCM-control (0 mM), Treh20 (9.5 mM), and Treh30 (13.4 mM). Chromatographic analysis of membrane lipids, carbohydrates, and polyols of the spermatozoa cytosol was performed. A decrease in the content of glycolipids in the plasma membranes of spermatozoa from 2.0% in native spermatozoa to 1.1–1.4% (frozen/thawed) and phospholipids from 71.2% (native) to 70.5% (frozen/thawed) reduced the progressive sperm motility from 65.7% in native spermatozoa to 12.6–27.6% (frozen/thawed). The same dynamics were observed for the viability parameter of 90.4% (native) and 27.0–41.2% (frozen/thawed). The Treh20 diluent, using a combination of fructose (36 mM) and trehalose (9.5 mM) saccharides, maximally preserved the lipid profile of plasma membranes and the composition of the cytosol of frozen/thawed rooster spermatozoa, which positively affected the indicators of general and progressive mobility and viability.

Propagation of plum (Prunus salicina L.) cultivar Frontier in vitro through control of shoot tip necrosis (STN) and validation of genetic integrity using ISSR markers

The present investigation was carried out to for developing a competent protocol for micropropagation of commercial plum cultivar Frontier, through control of shoot tip necrosis (STN) observed during in vitro shoot multiplication. Apical and axillary buds were collected from elite high yielding tree of Frontier and inoculated on Murashige and Skoog (MS) medium fortified with phytohormones at different concentrations. MS medium containing 0.5 µM BA, 0.5 µM Kin and 0.2 µM IBA resulted in highest shoot bud establishment (58.95%) after 4 weeks interval. However, all the shoots showed necrosis and died after 1st subculture. Necrosis appeared with yellowing of shoot tips within 7 days of first subculture and gradually increased downwards resulting in yellowing of leaves, followed by death of the shoots. Among various reported methods tested to control STN, combination of Fructose (0.50 mM) and Calcium chloride (1.0 mM) proved 100% effective. Highest in vitro shoot multiplication (fivefold) was recorded on medium enriched with 2.0 µM BA, 0.2 µM GA3, 0.2 µM IBA, 0.50 mM Fructose and 1.0 mM Calcium chloride. For rooting, two step rooting procedure was followed which resulted in 85.00% rooting when microshoots were given a dark treatment by dipping in ½ MS broth supplemented with 2.5 µM IBA for 48 h before transferring to semisolid ½ MS basal medium. Rooted plantlets when transferred in vivo for acclimatization, showed 70–80% survival after 10 weeks and no mortality on field transfer. Molecular analysis of the plants using ISSR primers proved 100% similarity between them and mother tree.

Fructose Increases the Expression of Uric Acid-Induced Oxidative Stress Genes, NOX4 and FOXO3, in Cultured HepG2 Cells

Abstract Objectives Uric acid is the final product of purine metabolism. The role of uric acid in oxidative stress is not clear. Studies have shown uric acid as antioxidant as well as pro-oxidant. High fructose sugar consumption increases uric acid levels by ATP depletion and the subsequent formation AMP which is a uric acid precursor. In our study, we investigated the effect of dose dependent treatments of fructose, uric acid, or a combination of both uric acid and fructose on expression of oxidative stress-related genes, mainly NADPH oxidase 4 (NOX4), superoxide dismutase 3 (SOD3), Forkhead Box O3 (FOXO3) and xanthine dehydrogenase (XDH) in cultured Hep G2 cells. Methods Human hepatocellular carcinoma [HEPG2] (ATCC HB8065) cells were treated with serum-free medium containing either 10 mM fructose, soluble uric acid (0.25 mM, 0.5 mM, or 0.75 mM), or a combination of fructose and uric acid. The cells were collected at the end of each incubation period (30 min, 2 and 24 hours) to extract total RNA. cDNA was synthesized from the extracted RNA. TaqMan assays were designed for use on real-rime PCR platform (QuantStudio 12 K Flex). TaqMan open array primers were custom-made by Thermo Fisher Scientific. Target quantification values were normalized against GAPDH levels and a combination of students’ t-test and ANOVA were applied. Results Soluble uric acid, either by itself or in conjunction with fructose, did not change the expression of the tested genes at 30 minutes or 2 hours. However, after 24 hours of incubation, uric acid increased the expression of NOX4 by 2 and FOXO3 by 1.5-fold (p < 0.05) whereas uric acid plus fructose-containing media increased the expression of NOX4 by 3.5 and FOXO3 by 2-fold (p < 0.05) after 24 hours of incubation as compared to control. No treatment differences were observed in the expression of SOD3. Conclusions These findings demonstrate that fructose increases the expression of uric acid-induced oxidative stress related genes. Funding Sources None.

Fructose levels are elevated in women with polycystic ovary syndrome with obesity and hyperinsulinemia.

Are fructose levels altered in women with polycystic ovary syndrome (PCOS)? Elevated serum fructose levels are associated with PCOS in Chinese Han women with overweight/obesity and hyperinsulinemia, and fructose levels are higher in follicular fluids from PCOS patients than from control subjects. Both fructose levels and PCOS are closely linked to obesity and insulin resistance. However, the relationship between fructose and PCOS remains largely unknown. A total of 157 Chinese Han women (67 controls and 90 PCOS patients) were recruited at Shengjing Hospital of China Medical University. To systematically study the relationship between serum fructose levels and PCOS, the study population of control subjects and PCOS patients was divided into overweight/obese and lean subgroups, and hyper-fasting serum insulin (FSI) and normal-FSI subgroups, respectively. Fructose levels were measured in serum samples collected from 80 patients with PCOS (32 lean, 48 overweight/obese) and 59 control subjects (27 lean, 32 overweight/obese) and in follicular fluid samples collected from mature follicles (17-22mm) and matched immature follicles (8-13mm) from 10 patients with PCOS and 8 control subjects. Serum fructose levels were increased in overweight/obese and hyper-FSI PCOS patients compared with the control subjects. Fructose had an area under the curve (AUC) of 79.7% at a cutoff value of 10.13pmol/μl, with a sensitivity of 91.7% and a specificity of 59.3% for the prediction of PCOS in overweight/obese patients. In the hyper-FSI group, fructose had an AUC of 72% at a cutoff value of 10.49pmol/μl, with a sensitivity of 71.1% and a specificity of 64.4% for the prediction of PCOS. There were no differences between fructose, total testosterone, free testosterone or dehydroepiandrosterone sulfate levels with respect to the reliability of predicting PCOS in the overweight/obese or hyper-FSI groups using the method outlined by Hanley and McNeil. Notably, the combination of fructose and total testosterone levels resulted in the highest AUC of 86.0% and high sensitivity (85.4%) and specificity (83.1%) for the prediction of PCOS in overweight/obese patients. The positive predictive value (PPV) and negative predictive value (NPV) were 80.4 and 87.5%, respectively. Similarly, the combination of fructose and total testosterone levels also resulted in a high AUC of 80.2% and moderate sensitivity (73.3%) and high specificity (84.7%) for the prediction of PCOS in hyper-FSI patients. The PPV and NPV were 78.6 and 80.6%, respectively. Furthermore, fructose levels were significantly higher in follicular fluids from PCOS patients than from control subjects, regardless of whether the follicles were mature or immature. It remains unclear whether fructose levels contribute directly to follicular development and the pathogenesis of PCOS or are merely a biomarker of these processes. The results of the present study, together with our previous study, show that monosaccharide status may be a novel marker for PCOS, highlighting the importance of further investigation into the role of monosaccharides, especially fructose, in the pathogenesis of PCOS. This work was supported by the National Natural Science Foundation of China (No. 81671423 and No. 81402130), the National Key Research and Development Program of China (No. 2018YFC1003100), Liaoning Provincial Key Research and Development Program (No. 2018225090), the Fok Ying Tung Education Foundation (No. 151039), Distinguished Talent Program of Shengjing Hospital (No. ME76) and Distinguished Teacher Program of China Medical University (No. QGZ2018079). No competing interests were declared.

SAT-149 The Metabolic Impairments Caused by Fructose and Prednisolone Intake Are Reversible After the Interruption of Exposure

Regular ingestion of fructose and continuous glucocorticoid (GC) treatment, separately, may result in metabolic adverse effects (1,2), but their combined impact is not yet understood. Thus, we aimed to evaluate the impact of a combination of fructose and prednisolone ingestion on glucose and lipid metabolism as well as the effect of the interruption of this combined treatment. Male Wistar rats were divided into five groups as follows: control (C) group, fructose (F) group (20% fructose in the drinking water), prednisolone (P) group (12.5 μg/mL prednisolone in the drinking water), fructose plus prednisolone (FP) group (a combination of fructose and prednisolone in the drinking water as for F and P groups) for 90 consecutive days. Half of the rats (group R) from the FP group were maintained for 90 more days with a discontinuation of fructose and prednisolone to monitor the parameters. The combination of fructose and GC intake led to an increase in the circulating triacylglycerol from the fourth week of treatment until the end of treatment (P<0.05, n=10). The FP group also exhibited higher abdominal adiposity and plasma insulin values, reduced insulin sensitivity, increased uricemia, and impaired hepatic redox balance that paralleled with augmented fat content in the liver (p<0.05, n=10). No major impact on blood glucose or glucose tolerance was observed in the FP group. These results observed in the FP group were associated with increased beta-cell mass (p<0.05, n=8). Moreover, the combination of sugar and GC treatment led to a reduction in the total hepatic protein kinase B (PKB) content and to a reduction in the phosphorylated 5’ AMP-activated protein kinase (AMPK) content in the epididymal adipose tissue (p<0.05, n=6). Combination of fructose and GC intake did not impact on proinflammatory protein contents in the adipose tissue. Complete cessation of treatments resulted in the normalization of plasma triacylglycerol and uric acid levels in the R group (p<0.05, n=10). Interruption of sugar and GC treatments also led to normalization of the peripheral insulin sensitivity, hepatic fat content and the hepatic redox state (p<0.05, n-10). We conclude that the combination of fructose and prednisolone intake induces a metabolic syndrome-like phenotype and that interruption of fructose and GC intake recover practically all metabolic impairments. These findings point to the risk of associating GC-based therapies with a regular intake of sweetened beverages and draws attention to the plasticity of the organism and the benefits of removing the causal factors, whenever possible. Reference: (1) Zhang et al., Nutrients. 2017 Mar 29;9(4). pii: E335. (2) Pasieka et al., Metabolites. 2016 Aug 3;6(3). pii: E24. Sources of Research Support: CNPq and CAPES.

Carbohydrates as binders in biomass densification for biochemical and thermochemical processes

This study was conducted to verify the potential application of carbohydrates with low molecular weight (MW) as ash-free as well as nitrogen- and sulfur-free binders in biomass densification for solid biofuels. In the first step, different kinds of carbohydrates including molasses (industrial byproduct), fructose, maltodextrin, sucrose, and glucose alongside lignosulfonate (phenolic byproduct from pulp and paper industry) were employed as binders in two sources of cellulosic biomass, namely spruce wood shavings and wheat straw. The results indicated that irrespective of the interaction of the factors, fructose and molasses worked as the best binders for spruce wood shavings and wheat straw biomass, respectively. In the second stage where interaction of the two selected binders with lubricants were studied, results showed that canola oil worked as the best lubricant for spruce wood shavings with the least negative effect on the impact resistance (durability) of the pelletized biomass. However, no lubricant resulted in a superior result with the least negative effect for wheat straw. Also, results in this stage indicated that pure glycerol, in contrast to crude glycerol, works as a binder rather than as a lubricant. In the third stage, the densification process at pilot-scale indicated that combination of molasses with crude glycerol worked as the best binder and lubricant for wheat straw biomass, while, a combination of fructose and canola oil worked as the best binder and lubricant for spruce wood shavings. In contrast to crude glycerol, pure glycerol proved to increase the friction in the pelletization unit and would not be an effective lubricant.

Biosynthesis of poly(3-hydroxybutyrate) and its copolymers by Yangia sp. ND199 from different carbon sources

A halophilic bacterium isolated from mangrove soil sample in Northern Vietnam, Yangia sp. ND199 was found capable of producing homopolymer poly(3-hydroxybutyrate) [P(3HB)], copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)], and copolymer poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] from different carbon sources. The presence of 3HB, 3HV, and 4HB monomers were confirmed by gas chromatography and nuclear magnetic resonance analysis. Only P(3HB) was produced using carbon sources such as fructose or by a combination of fructose with 1,5-pentanediol, 1,6-hexanediol, sodium hexanoate, or sodium octanoate. The biosynthesis of P(3HB-co-3HV) was achieved by adding cosubstrates such as sodium valerate and sodium heptanoate. When 1,4-butanediol, γ-butyrolactone or sodium 4-hydroxybutyrate was added to the culture medium, P(3HB-co-4HB) containing 4.0–7.1mol% 4HB fraction was accumulated. The molecular weights and thermal properties of polyesters were determined by gel permeation chromatography and differential scanning calorimeter, respectively. The results showed that Yangia sp. ND199 is able to produce polyester with high weight average molecular weight ranging from 1.3×106 to 2.2×106 Dalton and number average molecular weight ranging from 4.2×105 to 6.9×105 Dalton. The molecular weights, glass transition temperature as well as melting temperature of homopolymer P(3HB) are higher than those of copolymer P(3HB-co-3HV) or P(3HB-co-4HB).

Physical, bioactive and sensory quality parameters of reduced sugar chocolates formulated with natural sweeteners as sucrose alternatives

In this study, sugar alcohols, dietary fibers, syrups and natural sweeteners were used as sucrose alternatives in the production of reduced sugar chocolates (50% of cocoa parts) with enhanced bioactive profile. Formulated chocolates were evaluated for their physical (particle size distribution, texture) and sensory properties, sugar composition, polyphenolic compounds content and antioxidant capacity.All produced reduced sugar chocolates ensured >20% lower calorific value than conventional chocolate (prepared with sucrose). Formulated chocolates containing stevia leaves and peppermint exhibited the best sensory properties (especially with regard to mouthfeel, sweetness and herbal aroma), as well as the highest polyphenolic content and antioxidant capacity. Particle size and hardness of chocolates increased in comparison to conventional chocolate, in particular when the combination of fructose and isomalt or lactitol was used. The bioactive profile of produced chocolates was enriched with phenolic acids, flavone (luteolin and apigenin) and flavonol (quercetin) derivatives, which were not identified in control chocolate.

Melatonin and diet-induced metabolic syndrome in rats: impact on the hypophysial-testicular axis.

Abstract Combinations of fructose- and fat-rich diets in experimental animals can model the human metabolic syndrome (MS). In rats, the increase in blood pressure (BP) after diet manipulation is sex related and highly dependent on testosterone secretion. However, the extent of the impact of diet on rodent hypophysial-testicular axis remains undefined. In the present study, rats drinking a 10% fructose solution or fed a high-fat (35%) diet for 10 weeks had higher plasma levels of luteinizing hormone (LH) and lower plasma levels of testosterone, without significant changes in circulating follicle-stimulating hormone or the weight of most reproductive organs. Diet manipulation brought about a significant increase in body weight, systolic BP, area under the curve (AUC) of glycemia after an intraperitoneal glucose tolerance test (IPGTT), and plasma low-density lipoprotein cholesterol, cholesterol, triglycerides, and uric acid levels. The concomitant administration of melatonin (25 μg/mL of drinking water) normalized the abnormally high LH levels but did not affect the inhibited testosterone secretion found in fructose- or high-fat-fed rats. Rather, melatonin per se inhibited testosterone secretion. Melatonin significantly blunted the body weight and systolic BP increase, the increase in the AUC of glycemia after an IPGTT, and the changes in circulating lipid profile and uric acid found in both MS models. The results are compatible with a primary inhibition of testicular function in diet-induced MS in rats and with the partial effectiveness of melatonin to counteract the metabolic but not the testicular sequelae of rodent MS.

Production of Bacterial Cellulose from Gluconacetobacter persimmonis GH-2 using Dual and Cheaper Carbon Sources

Bacterial cellulose is an exopolysaccharide produced by various species of bacteria such as in the genera Gluconacetobacter, Agrobacterium, Achromobacter, Azotobacter, Rhizobium, Sarcina, Salmonella, Enterobacter, etc. In recent years, bacterial cellulose has been focused for the development of acoustic diaphragms, specialty membranes, biomedical wound care products, scaffold for tissue engineering, etc. In this study, effective culture method to produce bacterial cellulose from cheaper and dual carbon sources by Gluconacetobacter persimmonis was examined. Various fruit juices including pineapple, pomegranate, muskmelon, water melon, tomato, orange, and also molasses, starch hydrolyzate, sugarcane juice, coconut water, coconut milk were used as alternative carbon sources for bacterial cellulose production. Out of which muskmelon gave a highest cellulose yield of 8.08 g/L. Also glucose, fructose, sucrose, maltose, lactose, mannitol, inositol and glycerol were used in combination of two (1:1) as dual carbon sources. Out of these dual carbon sources, the combination of fructose and sucrose (1:1) gave the highest cellulose yield of 8.79 g/L. In this study, an attempt was made to reduce the cost of production medium for cellulose by using natural cheaper carbon sources.

Unhealthy diet and ultrafine carbon black particles induce senescence and disease associated phenotypic changes

Diet and pollution are environmental factors known to compromise “healthy aging” of the cardiovascular and respiratory systems. The molecular consequences of this permanent burden in these cells are still unknown. Therefore, this study investigates the impact of unhealthy diet on aging-related signaling pathways of human, primary cardiovascular cells and of airborne particles on lung epithelial and human endothelial cells. Nutrition health reports have shown that the diet in industrialized countries contains more than 100mg/dl low density lipoprotein (LDL) and a high fraction of added sugars, especially fructose. Several studies demonstrated that ultrafine particles can enter the circulation and thus may interact with endothelial cells directly. Both, dietary compounds and pollution derived particles, have been shown to increase the risk for cardiovascular diseases. To simulate an unhealthy diet, we supplemented cell culture media of human primary endothelial cells, smooth muscle cells and cardiomyocytes with LDL and replaced 1/3 of glucose with fructose. We observed hypertrophy in cardiomyocytes, enhanced proliferation in smooth muscle cells and increased senescence, loss of endothelial nitric oxide synthase and increased nuclear FoxO3A in endothelial cells. With respect to pollution we have used ultrafine carbon black particles (ufCB), one of the major constituents of industrial and exhaust emissions, in concentrations our lungs and vessels are constantly exposed to. These concentrations of ufCB increased reactive oxygen species in lung epithelial and vascular endothelial cells and reduced the S–NO content, a marker for NO-bioavailability, in endothelial cells. NO increases activation of Telomerase Reverse Transcriptase (TERT), an enzyme essential for telomere maintenance. TERT is required for proper endothelial cell function and is inactivated by Src kinase under conditions of oxidative stress. ufCB significantly increased Src kinase activation and reduced Telomerase activity in endothelial and lung epithelial cells. As a consequence, ufCB increased senescence of endothelial cells. To investigate whether ufCB show also effects in vivo, we instilled ufCB in concentrations not inducing inflammation into mice. Indeed, eNOS expression was reduced in the abdominal aorta of animals treated with ufCB.Thus, a combination of fructose and LDL in the diet and ufCB, as a major constituent of air pollution, seem to accelerate respiratory and cardiovascular cellular changes, which may compromise “healthy aging” and can lead to cardiovascular and pulmonary diseases.

Antioxidant Effects of Codonopsis pilosula Reverse Glucose Tolerance Impairment-Induced Testicular Dysfunction.

Glucose tolerance decline and insulin resistance often accompanied functional decline in multiple organs. The present study was to examine whether administration of Danseng (Codonopsis pilosula) root extract to adult male rats will reverse glucose tolerance impairment induced testicular dysfunction. The adult Sprague Dawley male rats were divided into three groups (n=6): control group, fructose alone group, fructose plus danseng group. In the fructose group, the tap drinking water was replaced by a 10% fructose solution for 8 weeks. All the rats administered with fructose got glucose tolerance impairment. In the fructose plus danseng group, after the adult male rats got glucose tolerance impairment, Danseng were added by 1 mg/kg body weight in their drinking water for another 4 weeks. Control rats were having standard chow and tap drinking water ad libitum. The rats were anaesthetized and blood was removed from the inferior vena cava and collected into heparinized tubes. The testis was excised and weighed, and used for biochemical analysis. Samples were kept frozen (-20°C) until assay. Activities of antioxidant enzymes in testis were measured by assay kit. The protein expression of steroidogenic acute regulatory protein (StAR), P450 side chain cleavage (P450scc), antioxidant enzymes, and protein kinase A (PKA) were detected by Western blot. Rats that received Danseng experienced a significant increase in blood testosterone level compared to the fructose group (p < 0.01). There were found no significant changes in weight of testis and the relative organ weight. No significant changes in testicular lipid peroxidation were observed between the three groups. The administration of fructose resulted in increased activity of superoxide dismutase (SOD) compared to control group (p<0.05). Combination of fructose and Danseng significantly enhanced the activity of SOD, catalase and glutathione peroxidase (GPx) compared to control group (p<0.05). A correspondent higher increase of testicular SOD expression was also observed in the fructose, and fructose plus Danseng group. The expression of StAR was also increased significantly in the fructose group compared with the control group (p<0.05). Combination of Danseng resulted in StAR expression recovered near the expression of control group. In conclusion, the obtained results strongly suggest that the consumption of fructose decreased the plasma testosterone level due to the declined testicular function, including imbalance activities of antioxidant enzymes, and resulted the compensatory increased StAR expression. The combination of Danseng has protective effects on testicular tissues and can reverse the effect of fructose on antioxidant enzymes. (poster)

Preservation of aerial conidia and biomasses from entomopathogenic fungi Beauveria brongniartii and Metarhizium anisopliae during lyophilization

In this study, we assessed the stability provided by different formulations to aerial conidia or biomasses (conidia, blastospores, and mycelia) of Beauveria brongniartii and Metarhizium anisopliae subjected to lyophilization. First, the impact of the freezing and drying processes on spore survival was evaluated. Whereas unprotected B. brongniartii spores showed high cryosensitivity, those of M. anisopliae were markedly harmed by the drying process. Then, the protective efficiency of 14 excipients was systematically evaluated and optimized regarding required concentrations. Fructose, glucose, and saccharose significantly enhanced viabilities for B. brongniartii and M. anisopliae spores following lyophilization, especially as a result of their cryoprotective effects. In addition, the effect of various bulking agents on spore survival was studied and dextran 4 was selected to enhance the physical properties of the lyophilized products. The combination of fructose and dextran 4 was further applied to prepare lyophilized biomasses of both fungi. In comparison to freshly harvested biomasses, the lyophilized products showed similar growth rates and a comparable production of virulent secondary metabolites such as destruxin A, destruxin B, or oosporein, suggesting their applicability as biological control agents.

Role of Transglycosylation Products in the Expression of Multiple Xylanases in Myceliophthora sp. IMI 387099

This study reports the regulation of multiple xylanases produced by Myceliophthora sp. IMI 387099. Fructose was found to positively regulate the expression of multiple xylanase when used as sole carbon source. The xylanases (EX(1 )and EX(2)) of acidic pI were expressed in the presence of simple sugars (glucose, arabinose, and xylose), whereas xylanase of both acidic as well as basic pI (EX(1,) EX(2,) EX(3), and EX(5)) were expressed in the presence of fructose, xylan, and combination of xylan and alcohol. The combination of fructose and xylan also led to expression of an additional xylanase (EX(4)). The positional isomer (iso-X4) was found to be the key transglycosylation product when cultures were grown in the presence of fructose and xylan. In the presence of alcohols, the higher expression of xylanase was ascribed to the synergistic effect of alkyl glycoside and other transglycosylation products present in the culture extracts.

Effect of fructose on motility, acrosome reaction and in vitro fertilization capability of boar spermatozoa

Aim: The present study was carried out to investigate the effects of fructose supplementation in glucose containing mTALP medium on motility, acrosome reaction and invitro fertilization capability of boar spermatozoa. Methods: Boar spermatozoa were preincubated, swum-up, resuspended and then incubated for 6h in mTALP medium supplemented with 0, 0.5, or 1.0mmol fructose in the presence of 5.0mmol glucose. After completion of the specified incubation period, motility was determined subjectively on the basis of speed of progression and on the type of forward movement of spermatozoa; acrosome status was evaluated by applying a triple staining technique; and invitro fertilization capability was assessed by acetic-orcein staining. Results: The combination of fructose and glucose (0.5+5.0mmol) supplements in mTALP medium improved sperm motility significantly (P<0.05), more than glucose alone (5.0mmol) at 2-6h of incubation. Acrosome reaction (live spermatozoa) and the sperm penetration rate was increased significantly (P<0.05) when the spermatozoa were treated with the combination of fructose and glucose compared with glucose alone, but the incidence of polyspermy was not significantly different between the treatments. Conclusion: These results suggest that the combination of glucose and fructose as supplements in mTALP medium improve the progressive motility, acrosome reaction and fertilization capability of boar spermatozoa. (Reprod Med Biol 2006; 5: 255-261).

Influence of various carbohydrates in shoot development in nodal culture of Guinean Anacardium occidentale genotypes

The influence of various carbohydrates in shoot development was studied in single nodal culture of Guinean cashew genotypes. Both apical and axillary nodal sections from one-and-half-year-old stock plants of elite selected cashews from Guinea-Bissau were used as explants. Six types of carbohydrates (sucrose, maltose, glucose, fructose, galactose and sorbitol) were tested separately at concentration of 83 mM. Sucrose, maltose and fructose showed the best performance and were additionally tested at concentrations of 0, 83 and 167 mM in order to evaluate the optimal concentration that promotes growth. These results indicated that the inclusion of a carbohydrate source is essential for shoot development. In the tested conditions the concentration of 83 mM was sufficient for shoot development since no significant differences were found when explants were cultured on a higher concentration. Moreover, the results showed that in spite of the concentration it is the type of carbohydrate that influences the shoot response. Maltose increased the number of developed shoots whereas fructose enhanced shoot length. These results suggested that a combination of these two carbohydrates could increase the yield of well-developed shoots in a single step. The effect of maltose and fructose was analysed separately and combined. The combination of fructose with maltose (each at 83 mM) promoted both the highest percentage of developed shoots and the highest shoot length when compared with the results obtained using these two sugars separately. With this study, we found a one-step carbohydrate combination that allowed overcoming the low yield of well-developed shoots observed in the current propagation system in Guinean cashew genotypes.

Transcriptional Regulation and Characterization of a Novel β-Fructofuranosidase-Encoding Gene from Bifidobacterium breve UCC2003

An operon involved in fructooligosaccharide breakdown was identified in the genome of Bifidobacterium breve UCC2003. This 2.6-kb transcriptional unit was comprised of three genes that encoded a putative permease, a conserved hypothetical protein, and a beta-fructofuranosidase. Active transcription of the operon was observed when B. breve UCC2003 was grown on sucrose or Actilight, while transcription appeared to be repressed when the organism was grown on glucose, fructose, a combination of glucose and sucrose, or a combination of fructose and sucrose. The beta-fructofuranosidase encoded by this operon was purified and biochemically characterized. The optimum pH and temperature for catalytic activity were determined to be pH 6.0 and 37 degrees C, respectively, and there was a dependence on bivalent cations, particularly manganese. The Km and Vmax values for sucrose hydrolysis were determined to be 25 +/- 2 mM and 24 +/- 3 micromol min(-1) mg(-1), respectively. Interestingly, the enzyme was shown to specifically catalyze cleavage of the beta(2-1) glycosidic bond between glucose and its neighboring fructose moiety in sucrose and other fructooligosaccharides with a relatively low degree of polymerization, and there was no detectable activity towards the beta(2-1) glycosidic bond between two fructose moieties within the same substrate. To our knowledge, such an enzymatic activity has not previously been described in bifidobacteria or other gram-positive bacteria.

Substituting Honey for Refined Carbohydrates Protects Rats from Hypertriglyceridemic and Prooxidative Effects of Fructose

Recent findings indicate that a high fructose diet has a prooxidant effect in rats compared with a starch diet. Because honey is rich in fructose, the aim of this study was to assess the effect of substituting honey for refined carbohydrates on lipid metabolism and oxidative stress. Rats were fed for 2 wk purified diets containing 65 g/100 g carbohydrates as wheat starch or a combination of fructose and glucose or a honey-based diet prepared by substituting honey for refined carbohydrates (n = 9/group). The same amount of fructose was provided by the honey and fructose diets. The hypertriglyceridemic effect of fructose was not observed when fructose was provided by honey. Compared with those fed starch, fructose-fed rats had a lower plasma alpha-tocopherol level, higher plasma nitrite and nitrate (NOx) levels and were less protected from lipid peroxidation as indicated by heart homogenate TBARS concentration. Compared with those fed fructose, honey-fed rats had a higher plasma alpha-tocopherol level, a higher alpha-tocopherol/triacylglycerol ratio, lower plasma NOx concentrations and a lower susceptibility of heart to lipid peroxidation. Further studies are required to identify the mechanism underlying the antioxidant effect of honey but the data suggest a potential nutritional benefit of substituting honey for fructose in the diet.