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The speedy growth of human population, the issues arising out of pollution, demand of invertase yield and dumping of wastes from agro-industry have been influenced to find new solutions for microbial enzymes. In this study, an attempt was made to produce the invertase by yeast isolated from honey bees. Influence of various process parameters like nutritional sources and environmental conditions was studied using waste material. The enzyme was precipitated by using the acetone to get the product for further experimental analysis. Subsequently, it was carried out for further purification by Sephadex G-50 column chromatography and dialysed. At that moment, optimization of culture conditions for the yield of invertase with agro waste was determined by process parameters. It was followed by the immobilization of microbial cells for increasing the yield of invertase by various matrices. In order to check the bioactive potential of invertase, the bread was prepared with and without the amendment of enzyme. The existing result could be supportive to future researchers for industrial applications of microbial invertase by immobilization approach. [ABSTRACT FROM AUTHOR]

This study investigated the synergistic effect of glass wool and sephadex sperm filtration techniques on improving the low semen quality of buffalo bulls with or without antioxidants. Semen was collected for 14 weeks, evaluated weekly, and divided into 12 groups. Good and low semen have extended using a Tris extender. Good semen quality is divided into three groups [S1: Tris extender; S2: Tris + vitamins E (2 mM) and C (5 mM); S3: Tris + glutamine (20 mM) and arginine (1 mM)]. Low semen quality was divided into three main groups and subdivided into three sub-groups (S4; Tris extender; S5: Tris +vitamins E (2 mM) and C (5 mM); S6: Tris + glutamine (20 mM) and arginine (1 mM)). In the 2nd and 3 rd main groups, glass wool and sephadex techniques are used with or without adding vitamins (E+C) and amino acids (Glutamine and arginine) and subdivided into three sub-groups with each technique, referred to as S7–S9 for glass wool and S10–S12 for sephadex techniques. Improving (P<0.01) motility, normal morphology, acrosome, and plasma membrane integrity and reducing total sperm abnormalities of low semen quality were noticed using sephadex with or without adding vitamins or amino acids post-cryopreservation. In conclusion, the sephadex technique removed immotile sperm, dead, abnormal, and good sperm harvested by this technique, which in turn may reflect positively on improving the pregnancy rate [ABSTRACT FROM AUTHOR]

The study compared efficacy of three sperm selection techniques in improving freezability of low-quality Murrah buffalo bull ejaculates. Sephadex (SEP), Sephadex ion-exchange filtration (SIE), and 40/80% BoviPure™(BP) gradient centrifugation protocols were standardized (ejaculates, n = 24). In Experiment-I, Sephadex G-75, G-100, and combined Sephadex G (75–100) column filtrates were compared. In Experiment-II, BP protocols: 200 g-10 min, 250 g-5, and 10 min, 300 g-10, and 15 min were compared. In fresh semen, Sephadex G (75–100) filtration and 250 g-5 min BP protocol improved sperm functions and were used in Experiment-III, where SEP G (75–100), SIE G (75–100), and 250 g-5 min BP processed ejaculates (n = 48) were cryopreserved and compared at post-thaw stage. The mean recovery rate differed in order: SEP > SIE > BP. SIE filtration significantly improved progressive motility, livability, membrane integrity, bovine cervical mucus penetration and live non-apoptotic sperm. Compared with control, all three techniques equally reduced post-dilution and post-thaw lipid peroxidation (LPO) rate. SEP post-thaw filtrates observed lower cryocapacitation-like changes, LPO (C11‐BODIPY581/591), and higher active mitochondria than other treatments. SIE and SEP equally improved post-thaw acrosome-intact sperm over BP. Filtration techniques, preferably, Sephadex ion-exchange filtration can most efficiently process low-quality buffalo bull ejaculates for cryopreservation and improve freezability. [ABSTRACT FROM AUTHOR]

Water in a crosslinked dextran gel, Sephadex® G25, is known to remain partially unfrozen during cooling and undergoes ice crystallization during rewarming. However, the mechanism of ice crystallization during rewarming is still unclear. To elucidate the frozen state of water in the gel, thermal behavior at low temperatures was investigated by using adiabatic calorimetry. Heat capacities and enthalpy-relaxation rates of the gel-containing water of mass ratio h (=g H2O/g dry G25) = 1.00 were measured between 80 and 300 K during rewarming, where the gel was intermittently heated at the rate of 50–100 mK min−1. Although an exotherm indicating ice crystallization during rewarming was confirmed with the gel precooled rapidly, at 5 K min−1, it disappeared when precooled slowly, at 20 mK min−1. During rewarming after the rapid cooling, two glass transitions were observed at ca. 175 K and 240–242 K. A higher-temperature glass transition due to the water trapped by the polymer network was not so clear, as it was overlapped with an endotherm due to the melting of small ice crystals, which indicates that the ice crystals formed have a broad size-distribution and that water in the gel is vitrified when ice crystals of even the smallest size cannot be formed. [ABSTRACT FROM AUTHOR]

The cow’s milk cheese whey was prepared by rennet coagulation, after obtaining the whey all its fats, proteins were removed then filtered through a 0.22 μm membrane filter. As a preliminary test to confirm the presence of glutathione (GSH), the colorimetric method using Ellman’s reagent (DTNB) was employed to detect GSH in the prepared whey. The yellow color, which was resulted from the reaction between GSH and DTNB can be easily recognized visually, indicated that GSH was present in the prepared whey. To emphasize the presence of glutathione in the prepared whey at a greater extent with a degree of more advanced stage, the HPLC technique was conducted using both C8 and C18 columns. It showed that GSH was eluted at 1.616 1.620 min respectively, corresponding to the standard glutathione sample. The concentration of GSH in the lyophilized prepared whey (without fats and whey portions) was determined according to the standard curve method and found to be equal to 3.1 μg Glutathione/1g of prepared whey powder. The purification of GSH from the prepared whey was done by using a Sephadex G-10 column. The presence and the amount of glutathione in the collected fractions were determined using the specific glutathione detection kit which gives the amount of glutathione in the collected fractions by units of micromoles. The maximum amount of glutathione was collected in fractions number 18, 19, and 20. The amount of glutathione in these fractions was 53.55, 81.77 and 55.23 μM, respectively. These three fractions were combined, the GSH quantity obtained from 1 g of prepared whey powder used in each run of gel filtration collectively produced a total amount of 190.55 μM of GSH (equal to 2.915 mg). In this study, we were able to detect GSH in cheese whey, quantify and purify. [ABSTRACT FROM AUTHOR]

We investigated the properties of the low molecular weight thermo-alkali-stable and mercury ion-tolerant xylanase production from Thermomyces dupontii KKU-CLD-E2-3. The xylanase was purified to homogeneity by ammonium sulfate, Sephadex G–100 and DEAE–cellulose column chromatography which resulted 27.92-fold purification specific activity of 56.19 U/mg protein and a recovery yield of 2.01%. The purified xylanase showed a molecular weight of 25 kDa by SDS–PAGE and the partial peptide sequence showed maximum sequence homology to the endo-1,4-β-xylanase. The optimum temperature and pH for its activity were 80 °C and pH 9.0, respectively. Furthermore, the purified xylanase can maintain more than 75% of the original activity in pH range of 7.0–10.0 after incubation at 4 °C for 24 h, and can still maintain more than 70% of original activity after incubating at 70 °C for 90 min. Our purified xylanase was activated by Cu2+ and Hg2+ up to 277% and 235% of initial activity, respectively but inhibited by Co2+, Ag+ and SDS at a concentration of 5 mM. The Km and Vmax values of beechwood xylan were 3.38 mg/mL and 625 µmol/min/mg, respectively. Furthermore, our xylanase had activity specifically to xylan-containing substrates and hydrolyzed beechwood xylan, and the end products mainly were xylotetraose and xylobiose. The results suggested that our purified xylanase has potential to use for pulp bleaching in the pulp and paper industry. [ABSTRACT FROM AUTHOR]

The electrochemical behavior of Sulfaclozine Sodium (SLC) was studied at a bare and sephadex-modified carbon paste electrodes by cyclic voltammetry and square wave voltammetry. The cyclic voltammetry (CV) showed a well-defined irreversible oxidation peak at 0.94 V in Britton- Robinson buffer pH 7.0. The strong affinity of SLC to sephadex allowed accumulation of SLC at the surface of electrode and thus higher electrochemical sensitivity to SLC. The influence of sephadex loading, the pH of the solution and the scan rate on the peak current was studied. A linear calibration curve covering the concentration range from 0.005 to 1 mM was obtained using SWV. The method was successfully applied for the determination of SLC in the veterinary pharmaceutical formulations with satisfactory accuracy and precision. [ABSTRACT FROM AUTHOR]

A proteolytic strains of Proteus vulgaris were isolated from 18 urinary tract infection (UTI) patients of both sexes, 10 were female (38.5%) and 16 from male (61.5 %), all strains produced IgA protease. The highly protease production was obtained by Proteusvulgaris number (16) of 27 mm in diameter after incubation for 18-24 hrs. at 37oC on skin milk agar medium and 0.16 mg/ml quantitatively by Bradford method. Isolation and purification of IgA protease from isolate number(16) on Sephadex G100 column which showed the presence of two peak of proteins and presence of one peak of enzyme. The molecular weight of the partially purified enzyme by gel filtration method in the 50KDa, there is a correlation between the clinical source strains and the protease molecular weight 50KDa. The purification fold by SDS-PAGE was (219.04) and (6.1) yield. The effect of the enzyme on the immunoglobulineswas studied and the results showed the enzyme have the abilities to digest IgA as detected by SDS-PAGE method and the IgG digestive was not clear. This study determine p.vulgaris infection activity by measurement levels of immunoglobulin A in 18 patients and 10 healthy as a control groups. The results showed significant decrease (P<0.05) in immunoglobulin A of p. vulgaris infection patients in compared to healthy group. [ABSTRACT FROM AUTHOR]

Abstract—The research cycle of thermal properties of water clusters in polysaccharide–water systems (starch and Sephadex G-100) with a low content of freezing water was continued based on the example of another modified polysaccharide, Sephadex G-25. This Sephadex has a more rigid three-dimensional structure compared with Sephadex G-100. The doublet structure of the melting curve of water clusters, which indicated the bimodal nature of their size distribution, was the main peculiarity of the data obtained for Sephadex G-25 by differential scanning calorimetry. The observed decrease in the temperature and heat of melting and crystallization of water clusters with a decrease in the humidity of Sephadex G-25, as in the case of other polysaccharides, was a typical manifestation of the size effect for nanosystems. At the same time, hysteresis occurred between the temperatures , and the heats of these transitions, which is also typical for small systems. It was found that the transformation processes (recrystallization, reorganization, and size change) in the system of clusters of crystallized supercooled water in Sephadex G-25 with low humidity during heating could occur both below the melting interval and inside it, which indicated the nonequilibrium state of the original set of nanoclusters. It was shown that the decrease of Vheat, as well as annealing within the melting interval led to a redistribution of clusters in size and, according to the revealed size effect, to an increase in the melting heat. These facts taken together indicated that the nonequilibrium state of the initially formed clusters during cooling and, as a consequence, their ability to transform with increasing temperature certainly play an important, if not a decisive, role in the manifestation of hysteresis in the thermal properties of water nanoclusters. [ABSTRACT FROM AUTHOR]

Abstract The aim of this study is to investigate the protective effect and underlying molecular mechanisms of isobavachalcone on Sephadex-induced lung injury in rats. The result showed isobavachalcone inhibited massive granulomas, decreased inflammatory cells infiltration and oxidative stress markers level, but it can increase antioxidant enzymes level. The ELISA assay exhibited isobavachalcone decreased TNF-α production in BALF and lung tissue. Western blotting analysis showed isobavachalcone can inhibit NF-κB pathway that may be mediated by upregulation of A20. Furthermore, we also found isobavachalcone can activate NRF2/HO-1 pathway and inhibit adhesion molecule expression. Taken together, the present results suggested that isobavachalcone can attenuate Sephadex-induced lung injury that may be related to inhibition of NF-κB mediated by upregulation of A20 and activation of NRF2/HO-1 signaling pathway. [ABSTRACT FROM AUTHOR]

Background: Phytic acid is the large reservoir of phosphorous of plant origin feed of monogastric animals. These animals are incapable of metabolizing phytic acid due to lack of phytase enzyme production. Phytases are a group of enzymes which are involved in hydrolyzing phytic acid and releasing inorganic phosphate Microorganisms are the main sources of phytases especially purified from filamentous fungi Aspergillus nigerS2. Materials and Methods: In the present study, a three step purification study of phytase from Aspergillus niger S2 was done by using Sephadex G-50. The active fractions pooled after chromatography step was used for molecular mass determination through sodium dodecyl sulphatepoly acrylamide gel electrophoresis (SDSPAGE). Phytase activity was determined at different pH between 3.0 and 8.0. The effect of temperature on enzyme activity was studied of temperature range of 30-60 °C. The action of the enzyme on several phosphorylated substrates was tested at concentrations of 3mM. The phytase enzyme function was tested on digestion of plant material. Results: The molecular mass of A. niger phytase S2was also found to be 50kDa. The enzyme showed optimal pH at 5.0. The enzyme showed maximum activity at 400C. The enzyme showed high substrate affinity towards sodium phytate. The purified enzyme activity on digestion of grass and hay showed positive results with increased protein content, reducing sugar and phosphate. Conclusion: The purified enzyme of the present study with these specifications can able to play a vital role in feed industry as a feed supplement in order to metabolize phytic acid and hence reduce soil pollution and eutrophication. [ABSTRACT FROM AUTHOR]

Abstract This study determined the optimal extraction conditions for ultrasonic-cellulase synergistic extraction of polysaccharides from pineapple pomace (PPP) using Plackett–Burman design and response surface methodology. The optimal hydrolysis temperature, ratio of material to water, pH value, hydrolysis time, ultrasonic power and the additive quantity of cellulase were 50 °C, 1:45 g/mL, 6.0, 100 min, 160 W and 2.0%, respectively, giving a extraction yield of 1.10 ± 0.03%. PPP was further isolated and purified by DEAE-52 cellulose and Sephadex G-100 chromatography columns, revealing four main elution peaks, named PPF0, PPF1, PPP2 and PPF3, were obtained. The molecular weight, monosaccharide compositions, structural features and appearance morphology of polysaccharide fractions (PPFs) were analyzed by high-performance liquid chromatography (HPLC), gel permeation chromatography (GPC), UV spectroscopy, fourier transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM). Furthermore, the hypoglycemic activities of PPFs with different concentrations were also investigated by insulin resistance HepG2 cells model in vitro. Results showed that PPF0, PPF1, PPF2 and PPF3 were composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, xylose, arabinose and fucose with molecular weight of 6.71 × 104, 1.11 × 104, 2.22 × 104 and 5.1 × 103 Da, respectively. All of them could alleviate the development of insulin resistance HepG2 cells with a dose-dependent relationship. The glucose consumption increased 46.4%, 50.5%, 82.1% and 53.6%; 86.8%, 81.6%, 86.8% and 84.2% at the concentration of 20 μg/mL, respectively, without or with insulin. These results suggested that PPFs can be explored as a potential hypoglycemic agent in biomedical and functional food. Highlights • The pineapple pomace as experimental materials, which can reduce environmental pollution and resources wasting. • The extraction conditions for pineapple pomace polysaccharides by ultrasonic-cellulase synergistic extraction were optimized. • Four main polysaccharides were isolated from pineapple pomace. • PPF0, PPF1, PPF2 and PPF3 exhibited hypoglycemic activity. [ABSTRACT FROM AUTHOR]

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Infertility has been defined as the disability to fulfill pregnancy after twelve months of regular sex coition, with no contraceptive taken. Male factor infertility was explained in terms of low spermatozoa concentration (oligozoospermia), or morphology (teratozoospermia), impaired motility (asthenozoospermia), or in some cases, total lack of spermatozoa in the ejaculate (azoospermia). In several cases, a collection of one, or more of these sperm variables defects may be noticed, evaluation these factors by the initial and most important step is seminal analysis. Our study aims to study some sperm characteristics in asthenozoospermia men in comparison with normozoospermia men before and after glass wool and Sephadex activation. This study involved 60 semen samples collected from male patients that came to male infertility clinic at Al-Nahrain University; the recruited semen samples were divided into 2 groups, (40 asthenozoospermic and 20 normozoospermic subjects). After collected semen samples, and assessed analysis of seminal fluid. Each semen samples were divided into 3 aliquots. The first aliquot prepared was In-Vitro to assessed sperm characterization before activation, the second part using glass wool filtration (GWF) technique, while the last part was prepared using Sephadex. Both techniques resulted equally in reducing sperm concentration, were equally effective in upgrading sperm motility and in minimizing round cell count; however, Sephadex was superior to glass wool method in upgrading the percentage of morphologically normal sperm. Both Sephadex and glass wool techniques has been proved effective to improve semen quality. [ABSTRACT FROM AUTHOR]

Abstract The optimal extraction conditions with yield of 11.86 ± 0.21% for extraction of crude polysaccharide from black mulberry fruit were obtained as extraction temperature 87 °C, extraction time 3 h and ratio of water to raw material 39 mL/g. Two purified fractions of BMP-1-1 and BMP-2-1 with average molecular weights of 615 and 405 kDa, respectively, were obtained by chromatography of DEAE-52 cellulose and Sephadex G-100. BMP-1-1 consisted of mannose, rhamnose, glucuronic acid, galactose and arabinose, BMP-2-1 consisted of rhamnose, glucuronic acid, galacturonic acid, galactose and arabinose. Crude BMP exhibited the strongest reducing power and protective effect on H 2 O 2 -induced oxidative injury in PC12 cells and the most effective scavenging activities against DPPH, ABTS and hydroxyl radical. [ABSTRACT FROM AUTHOR]

β-Galactosidase is a metal-activated enzyme, which breaks down the glucosidic bond of lactose and produces glucose and galactose. Among several commercial applications, preparation of lactose-free milk has gained special attention. The present objective is to demonstrate the activity kinetics of β-galactosidase purified from a non-pathogenic bacterium Arthrobacter oxydans SB. The enzyme was purified by DEAE-cellulose and Sephadex G-100 column chromatography. The purity of the protein was checked by high-performance liquid chromatography (HPLC). The purified enzyme of molecular weight ~ 95 kDa exhibited specific activity of 137.7 U mg−1 protein with a purification of 11.22-fold and yield 12.42 %. The exact molecular weight (95.7 kDa) of the purified protein was determined by MALDI-TOF. Previously, most of the studies have used Mg+2 as a cofactor of β- galactosidase. In this present investigation, we have checked the kinetic behavior of the purified β-galactosidase in presence of several bivalent metals. Lowest Km with highest substrate (ortho-nitrophenyl-β-galactoside or ONPG) affinity was measured in presence of Ca2+ (42.45 µM ONPG). However, our results demonstrated that Vmax was maximum in presence of Mn+2 (55.98 µM ONP produced mg−1 protein min−1), followed by Fe+2, Zn+2, Mg+2, Cu+2 and Ca+2. A large number of investigations reported Mg+2 as potential co factor for β-galacosidase. However, β-galactosidase obtained from Arthrobacter oxydans SB has better activity in the presence of Mn+2 or Fe2+. [ABSTRACT FROM AUTHOR]

Abstract In order to assist developing a natural, safe food-preservative, aqueous ethanolic extracts of leaves and berries of eight Finnish berry plants were fractionated with Sephadex LH-20 column chromatography. For each fraction, phenolic compounds were analyzed with NMR, UPLC-DAD-ESI-MS and HPLC-DAD. The antioxidant activities of the fractions were investigated using oxygen radical absorbance capacity (ORAC) assay, and the antibacterial activities were evaluated against foodborne pathogens Staphylococcus aureus and Escherichia coli. Antioxidant activities of the fractions correlated highly with both the total concentration and structural feature of phenolic compounds, including both flavonoids and non-flavonoid phenolics. ORAC value correlated strongly with the concentration of (+)-catechin, (−)-epicatecin, quercetin glycosides, and anthocyanins. Increase in size and number of sugar moieties may reduce the antioxidative activities of quercetin glycosides. Type of sugar moieties may have a significant role in influencing peroxyl-radicals scavenging ability of quercetin glycosides with monosaccharides as a single sugar moieties. Most of the fractions inhibited the target microbes. S. aureus strains expressed a higher sensitivity to phenolic compounds than E. coli strains. Graphical abstract Unlabelled Image Highlights • Food-grade extracts of berries and leaves were fractionated by column chromatography. • Phenolic compounds were identified and quantified thoroughly in each fraction. • Both antioxidative and antibacterial activities of fractions were investigated. • The contribution to bioactivities were compared among various phenolic compounds. • Results of the study will assist developing natural food preservatives. [ABSTRACT FROM AUTHOR]

Response surface methodology was used to optimize the extraction conditions for ultrasonic-assisted extraction of polysaccharides from mango pomace. The Optimum extraction conditions consisted of extraction temperature of 74 °C, ultrasonic power of 170 W, extraction time of 100 min, and raw material-to-water ratio of 1:40 g/mL. Under these conditions, the extraction yield was 3.71 ± 0.07%. Three novel polysaccharide fractions, MG-1, MG-2 and MG-3 were purified from the crude polysaccharides by using DEAE-52 cellulose and Sephadex G-100 column chromatography. The molecular weight and monosaccharide composition of polysaccharide fractions (MPFs) were analyzed by high performance liquid gel permeation chromatography (HPGPC) and HPLC analysis, respectively. The characterizations of MPFs were conducted with FT-IR, 1 H NMR and SEM. Furthermore, the anticancer activities of the polysaccharide fractions were also investigated in vitro . Results showed that MG-1, MG-2 and MG-3 exhibited significant anticancer activities against HepG2, MCF-7, A549, HeLa, A2780, HCT-116 and BGC-823 cells in a dose-dependent manner. MPFs were also showed to promote apoptosis as seen in the nuclear morphological examination study using calcein acetyl methoxy methyl easter (calcein-AM) and propidium iodide (PI) staining. This research could serve as a theoretical reference for the efficient utilization of MPFs in biomedical and functional food. [ABSTRACT FROM AUTHOR]

Nine antialgal active compounds, (i.e. trehalose ( 1 ), twenty-two methyl carbonate ( 2 ), (-)-dihydromenisdaurilide ( 3 ), 3,7,11,15-tetramethyl-2-hexadecen-1-ol ( 4 ), isophytol ( 5 ), 8-hexadecenol ( 6 ), 17-hydroxyheptadecanoic acid ( 7 ), trans-asarone ( 8 ) and 2-amino-3-mercaptopropanoic acid ( 9) ) were isolated from Ulva pertusa for the first time by sephadex LH-20 column chromatography, silica gel column chromatography and repeated preparative TLC. Except for compound 4 , all compounds represented novel isolated molecules from marine macroalgae. Further, antialgal activities of these compounds against Amphidinium carterae , Heterosigma akashiwo , Karenia mikimitoi , Phaeocystis globosa , Prorocentrum donghaiense and Skeletonema costatum were investigated for the first time. Results showed these nine compounds have selectivity antialgal effects on all test red tide microalgae, and antialgal activities against red tide microalgae obviously enhanced with the increase of concentration of antialgal compounds. Based on this, EC 50–96 h values of these nine compounds for six red tide microalgae were obtained for the first time. By analyzing and comparing EC 50–96 h values, it has been determined that seven compounds ( 1 , 3 , 4 , 6 , 7 , 8 and 9 ) showed the superior application potential than potassium dichromate or gossonorol and other six compounds as a characteristic antialgal agent against Heterosigma akashiwo , Karenia mikimitoi and Prorocentrum donghaiense . Overall this study has suggested that green algae Ulva pertusa is a new source of bioactive compounds with antialgal activity. [ABSTRACT FROM AUTHOR]

Contents: Our aim was to evaluate the effect of Sephadex filtration on respiratory activity of porcine spermatozoa and its relation with quality and functional sperm parameters. Samples were evaluated regarding oxygen uptake and sperm parameters: motility, plasma and acrosome membrane integrity, capacitation and acrosome reaction induction in vitro, plasma membrane functionality, determined by the hypo‐osmotic swelling test (HOST), and lipid peroxidation assessed by thiobarbituric acid assay. Sephadex filtration improved all routine quality parameters (motility, plasma and acrosome membrane integrity) and functional parameters (HOST, in vitro capacitation and true acrosome reaction levels) and produced a significant decrease in cryocapacitation and lipid peroxidation. Oxygen uptake increased in Sephadex samples (41 ± 7%) respect to single washing. Oxygen addition of carbonyl‐cyanide‐m‐chlorophenylhydrazone (CCCP) confirmed mitochondrial coupling in washed and Sephadex samples; showing an increase of 2.6 and 4.2 times for oxygen consumption in single washing and Sephadex ones, respectively. The increase in oxygen uptake with succinate addition with respect to basal oxygen uptake was significantly lower in Sephadex samples (63 ± 25%) than in the washed ones (183 ± 35%). Sephadex samples showed higher mitochondrial activity measured by oxygen consumption and improved quality and functional parameters. Our study recommends this protocol due to the fact that this filtration method removes dead or damaged spermatozoa allowing to obtain cryopreserved boar spermatozoa with optimized fertilizing capacity. [ABSTRACT FROM AUTHOR]

Density-gradients centrifugation (DGC) and filtration columns (FC) are used to separate deformed or dead sperm, debris, and other cells that may negatively affect the fertilizing capacity of sperm in fresh, chilled and frozen/thawed semen. The present study was conducted to evaluate the suitability of DGC (BoviPure®, Percoll® and Accudenz®) and FC (Sephadex G-15®) sperm selection procedures for fresh-extended and cold-stored ram semen by assessment of post-treatment sperm quality variables. Twenty normospermic ejaculates from ten adult Merino rams were used. Sperm concentration of recovered cells was greater ( P  < 0.001) after BoviPure treatment than other procedures in both fresh and cold semen. With the Sephadex method, there were more desirable values than with use of DGC procedures in several sperm motility variables measured by using the CASA system. In non-refrigerated semen samples, the percentage of progressive sperm motility (%PSM) after Sephadex filtration was greater ( P  < 0.05) than after BoviPure treatment; the straightline velocity (VSL) value after Sephadex filtration was greater ( P  < 0.01) than after Accudenz treatment; the amplitude of lateral head displacement (ALH) after Sephadex and Accudenz treatment was less than non-filtered semen ( P  < 0.001) and after Percoll ( P  < 0.01) and BoviPure ( P  < 0.05) treatments. In cold-stored semen samples, the %PSM after Sephadex filtration was greater than non-filtered ( P  < 0.05) semen and after BoviPure ( P  < 0.05), Percoll ( P  < 0.05) and Accudenz ( P  < 0.001) treatments. It is concluded that Sephadex column filtration can be used to select ram sperm in non-refrigerated and cooled semen, because percentage progressively motile sperm and some other sperm motility characteristics are greater with use of this techniques as compared with use of DGC methods. [ABSTRACT FROM AUTHOR]

Encapsulation and melanisation are innate immune reactions of insects against foreign intruders such as parasitoids. In an earlier study, we observed that immature life stages of the endoparasitoid Tranosema rostrale (Hymenoptera: Ichneumonidae) parasitizing Choristoneura fumiferana (Lepidoptera: Tortricidae) larvae experienced higher mortality due to encapsulation and melanisation when reared at high (30 °C) than at lower (10 °C, 20 °C) temperatures. Downregulation of T. rostrale polydnavirus genes in parasitized hosts and upregulation of two genes involved in the spruce budworm’s melanisation process were identified as likely contributors to parasitoid mortality at high temperature. However, levels of transcripts of genes involved in the spruce budworm’s cellular encapsulation process were not measured inasmuch as candidate genes, in the spruce budworm, had not yet been identified. In addition, our assessment of temperature-dependent encapsulation and melanisation of foreign objects in spruce budworm larvae was only partial. To fill these knowledge gaps, we injected Sephadex™ beads into unparasitized spruce budworm larvae and assessed their encapsulation/melanisation after the insects had been held at three different temperatures (10, 20, and 30 °C), and we identified spruce budworm genes putatively involved in the encapsulation process and quantified their transcripts at the same three temperatures, using a qPCR approach. As expected, both encapsulation and melanisation of Sephadex™ beads increased as a function of temperature. At the molecular level, three of the five genes examined ( Integrin β1 , Hopscotch , Stat92E ) clearly displayed temperature-dependent upregulation. The results of this study further support the hypothesis that a temperature-dependent increase in the encapsulation response of C. fumiferana against T. rostrale is due to the combined effects of reduced expression of polydnavirus genes and enhanced expression of host immune genes. [ABSTRACT FROM AUTHOR]

Sheep abomasum active proteases (SAPs) were purified by chromatography over a column of Sephadex G-25 and by ultrafiltration. The SAPs were found to contain proteases of molecular mass 14 kDa and active peptides with molecular masses varying in the range 651.39-4248.08 Da. Peptide fractions of SAPs reduced significantly (up to 72.0 ± 2.3% inhibition) accumulation of malondialdehyde (MDA) in brain homogenate of laboratory rats. [ABSTRACT FROM AUTHOR]

Contents: Routinely, swim‐up method is used to separate high‐quality sperm; however, long processing time and close cell‐to‐cell contact during the centrifugation step are inevitable elements of oxidative stress to sperm. The objective was to evaluate Sephadex™ and glass wool filtration to separate motile, intact and viable sperm for in vitro fertilization in buffalo. The cumulus–oocyte complexes (COCs) were collected from ovaries of slaughtered buffaloes by aspiration and matured for 24 hr in CO2 incubator at 38.5°C and 5% CO2. Matured COCs were rinsed twice in fertilization TALP and placed in the pre‐warmed fertilization medium without sperm. Cryopreserved buffalo semen was thawed at 37°C for 30 s and processed through Sephadex™, glass wool filtration and swim‐up (control). Total and motile sperm recovery rates were assessed, resuspended in fertilization TALP and incubated for 15–20 min in CO2 incubator. Samples prepared by each method were divided into two aliquots: one aliquot was studied for sperm quality (progressive motility, membrane integrity, viability, liveability), while the other was subjected to co‐incubation with sets of 10–15 in vitro matured oocytes. Data on sperm quality were analysed by ANOVA, while in vitro fertilizing rates were compared by chi‐squared test using SPSS‐20. Least significant difference (LSD) test was used to compare treatment means. Glass wool filtration yielded higher total and motile sperm recovery rate, while Sephadex™ filtration improved (p < .05) sperm quality (progressive motility, membrane integrity, viability, liveability). Sperm preparation through Sephadex filtration yielded higher in vitro fertilization rate in terms of cleavage rate compared to glass wool filtration and swim‐up (control). In conclusion, cryopreserved Nili‐Ravi buffalo sperm selected through Sephadex filtration showed improved quality and yielded better fertilization rates (cleavage rate) of in vitro matured/fertilized oocytes. Sephadex filtration could be a promising technique for use in in vitro fertilization in buffalo. [ABSTRACT FROM AUTHOR]

A polysaccharide (FVSP) was isolated from the base of Flammuliana Velutipes stipe, and FVSP was further purified by DEAE-cellulose-52 chromatography and Sephadex G-100 size-exclusion chromatography to obtain three fractions named FVSP-1, FVSP-2 and FVSP-3. Then their activation of macrophage cell RAW 264.7 and anti-proliferative effects to the murine melanoma B16F10 and fibroblasts L929 cells were evaluated by using the cell model experiments. The results indicated that the polysaccharide fractions could increase the proliferation and phagocytic activity of macrophage significantly and play an inhibited effect on the cancer cells. Moreover, the anti-proliferative activities of FVSPs increased with the participation of the antitumor factors induced from macrophage by polysaccharides fractions. Taken together, these results indicated that three polysaccharides fractions from the base of F. Velutipes stipe may be useful as potent antitumor agents for the prevention of tumorigenesis. [ABSTRACT FROM AUTHOR]

A new water-soluble polysaccharide named as AAPS-1, was extracted with warm water from the roots of Acanthophyllum acerosum and further purified by DEAE-cellulose A52 anion-exchange and Sephadex G-50 gel filtration chromatography. AAPS-1 was a neutral polysaccharide, with a molecular weight of 23.2 kDa and a specific optical rotation of +181.3° ( c 1.0, H 2 O), and comprised of Glucose, Galactose and Arabinose with a relative molar ratio of 1.6:5.1:1.0. Structure features of AAPS-1 were investigated by a combination of chemical and instrumental analysis, such as partial acid hydrolysis, methylation, periodate oxidation and Smith degradation, GC–MS, FTIR and NMR ( 13 C and 1 H). The data obtained indicate that AAPS-1 possessed a backbone of →6)-α- d -Gal p -(1→ residues, with branches attached to O -2 (∼35%) by α- d -Glc p -(1→ and at O -3 (∼16%) by α- d -Gal p -(1→ and by β- l -Ara p -(1 → 3)-β- l -Ara p -(1 → 3)-β- l -Ara p -(1→. The scavenging activity of AAPS-1 against DPPH radical was less than that of ascorbic acid at the same concentrations and the EC 50 value of AAPS-1 was 1.4 mg/ml. [ABSTRACT FROM AUTHOR]

The conversion of lignin into value-added aromatic monomers has attracted much attention as these monomers can be potentially used to prepare liquid fuel rather than heavy oil by further hydrodeoxygenation. A new process combining Sephadex and silica gel column chromatography was employed to obtain aromatic monomers with higher purity. It was found that pH and polarity of mobile phase were the most important factors affecting the separation effect for Sephadex G-10 and silica gel column chromatography, respectively. When the pH of mobile phase reached 10.5, the ionization of hydroxyl together with carboxyl groups resulted in electrostatic repulsion between the aromatic compounds and Sephadex G-10 gel, which counteracted the adsorption caused by hydrogen bond between aromatics and gel, causing the increase of the separation effect. Under the condition of pH above 10.5, Sephadex gel column chromatography gave 78.1% of purity of aromatic monomers (PAM) and 99.5% of recovery of aromatic monomers (RAM) due to the elimination of non-size exclusion effect. Silica gel column chromatography was further conducted to obtain 99.8% of PAM and 73.9% of RAM. The polar mobile phase was favorable for eluting aromatic monomers, while the non-polar mobile phase was beneficial to remove oligomers and increasing the PAM. [ABSTRACT FROM AUTHOR]

A novel transglutaminase (MsTGase) from Mythimna separata larvae was separated and purified; its biochemical property and enzymatic catalytic activities were investigated. MsTGase was obtained chromatographically by the precipitation of Sephadex G-100 gel and DEAE-Cellulose-52 ion-exchange column with 48-fold purification and a reproducible yield of approximately 12%. Molecular weight of the MsTGase was 63.5 K Da and its N -terminal amino acid sequence was GKIEEG-LVI. Michaelis constant of the MsTGase for the substrate N-CBZ-Gln-Gly was 12.83 m M with a V max of 7.99 U/mL. Optimum conditions for MsTGase activity were at 42 °C and pH7.5. The enzyme didn’t possess metal ion at its catalytic active site; its activity could be significantly inhibited by Mg 2+ , but activated by Ca 2+ . Chlorpyrifos and spinosad showed a strong potential to increase MsTGase activity, supporting the view that MsTGase was a novel target. Moreover, the formation of intermolecular cross-links of casein and bovine serum albumin polymerized by MsTGase in the presence of DTT was observed. These findings pave the way for future studies on the physiological role of MsTGase and the potential impact of its regulation on MsTGase-associated pest management. [ABSTRACT FROM AUTHOR]

A novel isolation strategy, online hyphenation of ultrasonic extraction, Sephadex LH-20 column chromatography combined with high-speed countercurrent chromatography, was developed for pure compounds extraction and purification. Andrographolide from Andrographis paniculata was achieved only in a single step purification protocol via the present strategy. The crude powder was ultrasonic extracted and extraction was pumped into Sephadex LH-20 column directly to cut the nontarget fractions followed by the second-dimensional high-speed countercurrent chromatography, hyphenated by a six-port valve equipped at the post-end of Sephadex LH-20 column, for the final purification. The results yielded andrographolide with the amount of 1.02 mg and a purity of 98.5% in a single step, indicating that the present method is effective to harvest target compound from medicinal plant. [ABSTRACT FROM AUTHOR]

A new water-soluble polysaccharide (ESPS-1), with the average molecular weight of 52.1 kDa and a specific optical rotation of +169.7° ( c 1.0, H 2 O), was successfully isolated by warm-water extraction and then purified using DEAE-Cellulose and Sephadex G-100 columns from the roots of Eremurus stenophyllus . The results from chemical and instrumental tests including FT-IR, methylation and GC–MS analysis, periodate oxidation and Smith degradation, partial acid hydrolysis, and NMR spectroscopy ( 1 H, 13 C, 1 H- 1 H COSY, HMQC and HMBC) demonstrated that ESPS-1 was a 2- O -acetylgalactan, composed of galactose, arabinose and mannose in a molar ratio of about 10:3:1, respectively. It has a backbone of (1 → 6)-α- d -galactopyranosyl residues, with branches at O -3 consisting of α- d -Man p -(1 → 3)-α- l -Ara f -(1 → 3)-α- l -Ara f -(1 → 3)-α- l -Ara f -(1→. The acetyl groups are substituted at O -2 of (1 → 6)-linked Gal p and the degree of acetylation was about 14%. [ABSTRACT FROM AUTHOR]

Streptococcus mutans is recognized as one of the key contributors to the dysbiotic state that results in dental caries. Existing treatment strategies reduce the incidence of tooth decay, but they also eliminate both the cariogenic and beneficial microbes. Here we introduce a novel treatment alternative using Sephadex, cross-linked dextranomer microspheres (DMs), typically used for gel filtration chromatography. In addition DM beads can be used for affinity purification of glucosyltransferases (GTFs) from S. mutans. In this study we take advantage of the native pathogenic mechanisms used by S. mutans to adhere, form a biofilm and induce dental caries through the expression of surface-associated GTFs. We demonstrate that planktonic and biofilm-grown (adhered to hydroxyapatite-coated pegs to mimic the tooth surface) S. mutans, specifically and competitively attach to DMs. Further investigation demonstrated that DMs are a specific affinity resin for S. mutans and other cariogenic/pathogenic oral streptococci, whereas other commensal and probiotic strains failed to readily adhere to DMs. Using antimicrobial cargo loaded into the DM lumen, we demonstrate that when in co-culture with non-binding to even modestly binding commensal species, S. mutans was selectively killed. This proof of concept study introduces a novel means to safely and effectively reduce the pool of S. mutans and other pathogenic streptococci in the oral cavity with limited disturbance of the necessary commensal (healthy) microbiota when compared with current oral healthcare products. [ABSTRACT FROM AUTHOR]

Eugenia dysenterica ex DC Mart. (Myrtaceae) is a Brazilian tree with pharmacological and biological properties. The aqueous leaf extract, rich in polyphenols, was tested in the human neuroblastoma cell line SH-SY5Y to evaluate its effect on cell viability. The extract and two isolated compounds were also assessed for the potential inhibitory activity on acetylcholinesterase, an enzyme related to Alzheimer's disease. A simple chromatographic method using Sephadex LH-20 was developed to separate catechin and quercetin from the aqueous leaf extract of E. dysenterica . Identification was carried out by spectroscopic techniques IR, UV, and 1 H and 13 C NMR. The IC 50 values were obtained by constructing dose-response curves on a graph with percentage inhibition versus log of inhibitor concentration and compared with physostigmine, a well-known AChE inhibitor. The extract was toxic for SH-SY5Y cells at concentrations higher than 7.8 μg/ml given for 24 h. The decline in SH-SY5Y cell viability appears to be related to its antiproliferative activity. The extract also showed relatively moderate acetylcholinesterase inhibitory activity of 66.33% ± 0.52% at 1.0 mg/ml with an IC 50 value of 155.20 ± 2.09 μg/ml. Physostigmine, quercetin, and catechin showed IC 50 values of 18.69 ± 0.07, 46.59 ± 0.49, and 42.39 ± 0.67 μg/ml, respectively. [ABSTRACT FROM AUTHOR]

Proteases from viscera of Silver carp (Hypophthalmichthys molitrix) fish have been extracted and characterized. The alkaline proteases show optimum activity in 0.2M Tris-HCl buffer at pH 8.5 and 45°C using soluble milk casein as substrate. The crude alkaline protease lost about 35% or 51% of its specific activity if it was heated at 50C or kept at pH 9 for 60 min., respectively. Purification of proteases purified by ammonium sulfate precipitation, gel-filtration using Sephadex G 50 column and ultrafiltration via polyamide membrane (30 KDa) led to increase its specific activity up to 20, 24 and 99 fold, respectively. [ABSTRACT FROM AUTHOR]

A novel on-line comprehensive two-dimensional liquid chromatography (2D-LC) method by coupling mixed-mode gel liquid chromatography (MMG-LC) with reversed phase liquid chromatography (RPLC) was developed. A mixture of 17 reference compounds was used to study the separation mechanism. A crude water extract of Flos Carthami was applied to evaluate the performance of the novel 2D-LC system. In the first dimension, the extract was eluted with a gradient of water/methanol over a cross-linked dextran gel Sephadex LH-20 column. Meanwhile, the advantages of size exclusion, reversed phase partition and adsorption separation mechanism were exploited before further on-line reversed phase purification on the second dimension. This novel on-line mixed-mode Sephadex LH-20 × RPLC method provided higher peak resolution, sample processing ability (2.5 mg) and better orthogonality (72.9%) versus RPLC × RPLC and hydrophilic interaction liquid chromatography (HILIC) × RPLC. To the best of our knowledge, this is the first report of a mixed-mode Sephadex LH-20 × RPLC separation method with successful applications in on-line mode, which might be beneficial for harvesting targets from complicated medicinal plants. [ABSTRACT FROM AUTHOR]