HPLC method with fluorescence detection for the quantitative determination of flaxseed lignans

We report a rapid and simple HPLC method with fluorescence detection for the quantification of the major flaxseed lignan, secoisolarisiresinol diglucoside (SDG) and its major metabolites. The method is specific for SDG, secoisolarisiresinol (SECO), enterodiol (ED) and entrolactone (EL) in rat serum. The assay procedure involves chromatographic separation using a Waters Symmetry C₁₈ reversed-phase column (4.6 mm × 150 mm, 5 μm) and mobile phase gradient conditions consisting of acetonitrile (0.1% formic acid) and water (0.1% formic acid). SDG extraction from serum requires the use of Centrifuge filters while SECO, ED and EL are extracted with diethyl ether. The organic layer is evaporated and reconstituted in 100 μL of mobile phase and 50 μL of reconstituted sample or filtrate is injected onto the column. Total run time is 25 min. Calibration curves are linear (r² ≥ 0.997) from 0.05 to 10 μg/mL for SDG and EL and 0.01–10 μg/mL for SECO and ED. Precision and accuracy are within USFDA specified limits. The stability of all lignans is established in auto-injector, bench-top, freeze–thaw and long-term stability at −80 °C for 30 days. The method's reasonable sensitivity and reliance on more widely available HPLC technology should allow for its straightforward application to pharmacokinetic evaluations of lignans in animal model systems such as the rat.     

Production and partial purification of invertase using <Emphasis Type="Italic">Cympopogan caecius</Emphasis> leaf powder as substrate

The present study investigates the efficiency of Aspergillus niger to produce invertase, an industrially important enzyme by using powdered stem of Cympopogan caecius (Lemon grass) as sole substrate and sole carbon source for the microorganism. The molecular weight of invertase was estimated to be 66–70 kDa by sodium do decyl sulphate poly acrylamide gel electrophoresis (SDS PAGE). The production of the enzyme was studied at different pH scales ranging from pH 4.0 to 7.0 at a constant temperature of 30°C and 2% substrate concentration. The maximum production of invertase (specific activity −0.0516 μk/mg protein) was obtained at pH 5.5 at 30°C temperature, and incubation for 48 h. The activity was found to be stable at pH 5.5 for 30 min. The enzyme was found to be stable in the temperature range of 20–55°C. The effect of divalent metal ions Cu²⁺, Fe²⁺, Co²⁺ on the activity of the enzyme invertase showed that these ions affected the activity by a certain factor. The study can be further industrially exploited in a country-like India where lemon grass is found in plenty and can be used as substrate for enzyme production. Moreover, the preparation of the substrate is also a simple process.     

Enzymatic bleaching of kraft pulp by xylanase from Aspergillus sydowii SBS 45

Crude xylanase from Aspergillus sydowii SBS 45 was tested for enzymatic bleaching of kraft (Decker) pulp. After optimization of three parameters, consistency of pulp, retention time and enzyme dose, considerable increase in the release of UV and visible absorbance spectra of materials and reducing sugars was observed, which clearly indicated the action of xylanase on pulp. Final brightness of pulp was increased from 29.42 to 70.42% and kappa number was reduced from 15.93 to 1.61, when 25 U of xylanase was given with a retention time of 5 h and at a consistency of 10%. When 10 U g⁻¹ xylanase was given, 14.3% elemental chlorine and 14.3% H₂O₂ could be reduced and when 25 U g⁻¹ xylanase was given 14.3% elemental chlorine and 28.6% H ₂O₂ could be reduced thereby retaining the brightness at control level.     

Ethanol fermentation of mahula (<Emphasis Type="Italic">Madhuca latifolia</Emphasis>) flowers using free and immobilized bacteria <Emphasis Type="Italic">Zymomonas mobilis</Emphasis> MTCC 92

Mahula (Madhuca latifolia L.) is a deciduous tree commonly found in the tropical rain forests of Asian and Australian continent. Corolla, the edible part of its flowers, is rich in fermentable sugar (37 ± 0.23%; on dry weight basis). Batch fermentation of mahula flowers was carried out using Zymomonas mobilis MTCC 92 free cells and cells immobilized in calcium alginate matrix. The ethanol productions were 122.9 ± 0.972 and 134.6 ± 0.104 g/kg flowers on dry weight basis using free and immobilized cells, respectively, after 96 h of fermentation, which showed that cells entrapped in calcium alginate matrix yielded 8.7% more ethanol than free cells. Further, the immobilized cells were physiologically active up to three more cycles of fermentation producing 132.7 ± 0.095, 130.5 ± 0.09 and 128.7 ± 0.056 g ethanol per kg flower in first, second and third cycle, respectively.     

Inhibition of angiogenesis- and inflammation-inducing factors in human colon cancer cells <Emphasis Type="Italic">in vitro</Emphasis> and <Emphasis Type="Italic">in ovo</Emphasis> by free and nanoparticle-encapsulated redox dye,DCPIP

Background  

The redox dye, DCPIP, has recently shown to exhibit anti-melanoma activity in vitro and in vivo. On the other hand, there is increasing evidence that synthetic nanoparticles can serve as highly efficient carriers of drugs and vaccines for treatment of various diseases. These nanoparticles have shown to serve as potent tools that can increase the bioavailability of the drug/vaccine by facilitating absorption or conferring sustained and improved release. Here, we describe results on the effects of free- and nanoparticle-enclosed DCPIP as anti-angiogenesis and anti-inflammation agents in a human colon cancer HCT116 cell line in vitro, and in induced angiogenesis in ovo.     

Adenovirus stimulates choline efflux by increasing expression of organic cation transporter-2

We examined the effect of wild-type human adenovirus (Ad5) on choline transport in murine lung epithelia (MLE) and in rodent primary alveolar type II cells. Cells were active in pH-sensitive, reversible transport of choline, a process blocked pharmacologically with phenoxybenzamine, an inhibitor of organic cation transporters (OCT). PCR products for the choline transporters, OCT-1 and OCT-2, were detected, but only OCT-2 protein was robustly expressed within MLE and primary alveolar epithelial cells. Ad5 produced a two- to threefold increase in choline efflux from cells, resulting in a significant reduction in intracellular choline content and its major product, phosphatidylcholine. Effects of Ad5 on choline efflux were inhibited with phenoxybenzamine, and choline efflux was attenuated by OCT-2 small interfering RNA. Adenovirus also produced a dose-dependent increase in immunoreactive OCT-2 levels concomitant with increased cellular OCT-2 steady-state mRNA. These results indicate that adenoviruses can significantly disrupt choline trafficking in lung epithelia by upregulating expression of an alveolar protein involved in organic cation transport.     

Impact of Juvenile Stocking Size on the Major Carp Production in a Minor Reservoir,Bibinagar, India

The present study deals with the impact of juvenile stocking sizes of the major carp on production in a minor reservoir, Bibinagar, Nalgonda, India. Reservoir surface area equaled over 23.8 ha and water from the reservoir was used for irrigation and fisheries. Four experiments were conducted for 4 years from 2000–2001 to 2003–2004. The experiments were planned in such a way that every year juvenile stocking size was held constant and subsequent fish production analyzed. During the first year’s experiment (2000–2001) a stocking size of 25–30 mm (fry) was maintained. Similarly 50–55 (advanced fry), 75–80 mm (fingerling) and 100–105 mm (advanced fingerling) were stocked during 2001–2002, 2002–2003 and 2003–2004 respectively. Uniform yearly stocking densities (2000/ha) were established in each experiment in the month of July and fish were harvested in June of the subsequent year. Major carp production was enhanced with larger stocking size. Yearly productions equaled 144.00, 231.48, 632.91 and 1005.03 kg/ha/year with the above stocking sizes. Variation in stocking sizes had significant (P<0.05) effects on fish production. The number of fish per kg production decreased gradually with increases in stocking size with an average value of 3.97. Reservoir production of catla was the most abundant species, followed by rohu, common carp, mrigal and grass carp. These results show that stocking size has a great impact on fish production and the stocking of advanced fingerlings will provide maximum carp production in minor reservoirs in India.     

The sodium current underlying action potentials in guinea pig hippocampal CA1 neurons

Neurons were acutely dissociated from the CA1 region of hippocampal slices from guinea pigs. Whole-cell recording techniques were used to record and control membrane potential. When the electrode contained KF, the average resting potential was about -40 mV and action potentials in cells at -80 mV (current-clamped) had an amplitude greater than 100 mV. Cells were voltage-clamped at 22-24 degrees C with electrodes containing CsF. Inward currents generated with depolarizing voltage pulses reversed close to the sodium equilibrium potential and could be completely blocked with tetrodotoxin (1 microM). The amplitude of these sodium currents was maximal at about -20 mV and the amplitude of the tail currents was linear with potential, which indicates that the channels were ohmic. The sodium conductance increased with depolarization in a range from -60 to 0 mV with an average half-maximum at about -40 mV. The decay of the currents was not exponential at potentials more positive than -20 mV. The time to peak and half-decay time of the currents varied with potential and temperature. Half of the channels were inactivated at a potential of -75 mV and inactivation was essentially complete at -40 to -30 mV. Recovery from inactivation was not exponential and the rate varied with potential. At lower temperatures, the amplitude of sodium currents decreased, their time course became longer, and half-maximal inactivation shifted to more negative potentials. In a small fraction of cells studied, sodium currents were much more rapid but the voltage dependence of activation and inactivation was very similar.