Recovery of squalene from wine lees using ultrasound assisted extraction-a feasibility study

The present work is a systematic approach for valorization of wine lees regarding the recovery of squalene, a bioactive lipid. Such a study is presented for the first time in literature. Separate examination of squalene content in "light" and "heavy" lees from different vinification processes by RP-HPLC demonstrated that these waste streams can be used as a source for this lipid, despite variations due to technological or genetic effects. Next, ultrasound assisted extraction of squalene from the "industrial waste" (the mixture of wine lees generated from different wines) using n-hexane was optimized with the aid of response surface methodology (independent variables: sonication duration and duty cycles). Autolysis was monitored through optical microscopy. Squalene yield (0.6 ± 0.08 g SQ/kg dry lees) was comparable to that of recently examined potential sources (0.2-0.35 g SQ/kg dry olive pomace and 0.06 g SQ/kg olive leaves).     

Caffeic acid derivatives production by hairy root cultures of Echinacea purpurea

Inoculation of leaf explants of Echinacea purpurea (Moench) with Agrobacterium rhizogenes induced hairy roots with the capacity to produce biologically active caffeic acid derivatives (CADs), especially cichoric acid. The kinetics of growth, the uptake of macronutrients, and the accumulation of CADs were investigated in heterotrophically cultured hairy roots for a 50 day period. A maximum of 12.2 g L(-1) dry biomass was achieved in MS nutrients supplemented with 30 g L(-1) sucrose on day 40. The mathematical relationship between hairy root growth and conductivity was established during the exponential phase in Erlenmeyer flasks. HPLC analyses of methanolic (0.1% phosphoric acid; 70:30, v/v) extracts from hairy roots revealed the presence of important CADs: cichoric acid (19.21 mg g(-1) dry biomass), caftaric acid (3.56 mg g(-1) dry biomass), and chlorogenic acid (0.93 mg g(-1) dry biomass). These results demonstrate that biotechnological production of CADs in hairy roots of E. purpurea is possible. Furthermore, these hairy root cultures offer, for the very first time, an excellent biological model to study the biosynthetic pathway of medicinally important CADs.     

Industrial glycerol as a supplementary carbon source in the production of beta-carotene by Blakeslea trispora

The dynamics of industrial types of glycerol as a supplementary carbon source to glucose for beta-carotene production by Blakeslea trispora was investigated in batch cultures. The growth kinetics, cellular lipid accumulation-degradation, substrate assimilation, and beta-carotene production were clearly dependent on the level of addition of pure glycerol. The highest beta-carotene production (15.0 mg/g of dry biomass) was obtained at an initial glycerol concentration of 60.0 g/L. Substitution of pure glycerol by the nonpurified soap byproduct did not inhibit cell growth. Conversely, partial purification of the biodiesel byproduct by removing methanol and fatty acids was unavoidable for cell growth. Both types of industrial glycerol stimulated beta-carotene synthesis more than 10 (soap byproduct) and 8 times (biodiesel byproduct) compared to control medium. The maximum beta-carotene contents were 10 and 8 mg/g of dry biomass, respectively, and its relative content in the carotenoid fraction was 86-88%.     

Effect of Foliar Salicylic Acid Applications on Growth,Chlorophyll, and Mineral Content of Cucumber Grown Under Salt Stress

The objective of this study was to determine the effect of foliar salicylic acid (SA) applications on growth, chlorophyll, and mineral content of cucumber grown under salt stress. The study was conducted in pot experiments under greenhouse conditions. Cucumber seedlings were treated with foliar SA applications at different concentrations (0.0, 0.25, 0.50, and 1.00 mM). Salinity treatments were established by adding 0, 60, and 120 mM of sodium chloride (NaCl) to a base complete nutrient solution. The SA was applied with spraying two times as before and after transplanting. Salt stress negatively affected the growth, chlorophyll content and mineral uptake of cucumber plants. However, foliar applications of SA resulted in greater shoot fresh weight, shoot dry weight, root fresh weight, and root dry weight as well as higher plants under salt stress. Shoot diameter and leaf number per plant increased with SA treatments under salt stress. The greatest chlorophyll content was obtained with 1.00 mM SA treatment in both saline and non-saline conditions. Leaf water relative content (LWRC) reduced in response to salt stress while SA raised LWRC of salt stressed cucumber plants. Salinity treatments induced significant increases in electrolyte leakage. Plants treated with foliar SA had lower values of electrolyte leakage than non-treated ones. In regard to nutrient content, it can be interfered that foliar SA applications increased almost all nutrient content in leaves and roots of cucumber plants under salt stress. Generally, the greatest values were obtained from 1.00 mM SA application. Based on these findings, the SA treatments may help alleviate the negative effect of salinity on the growth of cucumber.     

GENOTYPIC VARIATIONS IN POTASSIUM UPTAKE AND UTILIZATION IN COTTON

Genotypic differences in potassium (K) uptake and utilization were compared for eight cotton cultivars in growth chamber and field experiments. Four of the cultivars (‘SGK3’, ‘SCRC18’, ‘SCRC21’ and ‘SCRC22’) typically produce lower dry mass and the other four (‘Nannong8’, ‘Xiangza2’, ‘Xinluzao12’ and ‘Xiangza3’) produce greater dry mass in K-deficient solution (0.02 mM). The mean dry weight of seedlings (five-leaf stage) of cultivars with greater biomass was 155% higher than that of cultivars with lower biomass yield under K deficiency. However, all the genotypes had similar dry matter yields in K-sufficient solution (2.5 mM). Thus, the four cultivars with superior biomass yield under low K medium may be described as K efficient cultivars while the inferior cultivars may be described as K inefficient. Although seeds of the studied cultivars originated from different research institutes or seed companies, there were little differences in seed K content among them, irrespective of their K efficiency. Consequently, there were no significant differences in K accumulation in seedlings (4 d after germination in a K-free sand medium) just before transferring to nutrient solutions. However, the K efficient genotypes, on average, accumulated twice as much K at 21 d after transferring to K-deficient solution (0.02 mM). A much larger root system as well as a slightly higher uptake rate (K uptake per unit of root dry weight) may have contributed to the higher net K uptake by the K efficient cultivars. In addition, the K efficiency ratio (dry mass produced per unit of K accumulated) and K utilization efficiency (dry mass produced per unit of K concentration) of the K efficient cultivars exceeded those of the K inefficient genotypes by 29% and 234%, respectively, under K deficiency. On average, the K efficient cultivars produced 59% more potential economic yield (dry weight of all reproductive organs) under field conditions even with available soil K at obviously deficient level (60 mg kg^?1). We noted especially that the four K inefficient cultivars studied were all transgenic insect-resistant cotton, suggesting that the introduction of foreign genes (Bt and CpTI) may affect the K use efficiency of cotton.     

Effect of exogenous application of salicylic acid on salt-stressed sorghum growth and nutrient contents

This study was conducted to investigate the effect of salinity and foliar application of salicylic acid (SA) on sorghum biomass and nutrient contents. Treatments were comprised of salinity levels (0 and 100?mM NaCl) and SA concentrations (0.3, 0.7, 1.1 and 1.5?mM). Salinity increased sodium (Na), chlorine (Cl) and copper (Cu) but decreased nitrogen (N), phosphorus (P), potassium (K), magnesium (Mg), sulfur (S), iron (Fe), zinc (Zn) and manganese (Mn) contents and the root and shoot dry matter. Fe and Zn were the most affected nutrients by salinity. However, SA reduced Na and Cl but increased plant dry matter and nutrient content. SA had greater positive effects on root than on shoot dry matter. Maximum increases through SA were achieved in N, K, Fe, Mn, Cu, and shoot weight under salt stress but in Zn and root weight under non-saline condition. In most cases 1.1?mM was the most effective SA concentration in reducing the negative effects of salinity.     

Protein utilization during soybean tempe fermentation.

The aim was to identify to what extent proteins were utilized during the fermentation of bacteria-free tempe prepared with acidified soybean cotyledons and Rhizopus oligosporus NRRL 2710 at 30 degrees C. Dry matter declined continuously during the fermentation to 980 g/(kg of initial dry cotyledons) at 28 h, 910 g at 46 h (when the tempe was judged mature), and 835 g at 72 h. The decrease in dry matter was due mainly to reduction in crude lipid, amounting to 65 g/(kg of initial dry cotyledons) at 46 h and 135 g/(kg of initial dry cotyledons) at 72 h and representing approximately 70% and 80%, respectively, of the total dry matter loss. Protein oxidation (estimated from ammonia production) was 5 g at 28 h,10 g at 46 h, and 20 g/(kg of initial dry cotyledons) at 72 h. The total amount of soy protein hydrolyzed, including that incorporated into mold biomass, was estimated to be 80 g/(kg of initial dry cotyledons) at 28 h incubation, 95 g at 46 h, and 100 g at 72 h. The hydrolyzed protein at 46 h represented 25% of the initial protein. Of this hydrolyzed protein, it is suggested that approximately 65% remained in the tempe as amino acids and peptides, 25% was assimilated into mold biomass, and 10% was oxidized.     

Production of organically bound selenium yeast by continuous fermentation

This paper describes a protocol for incorporation of sodium selenite or sodium selenate into Saccharomyces cerevisiae biomass by continuous fermentation in a medium with minimal sulfur and methionine concentrations. Selenium incorporation was followed by atomic absorption analysis and methylene blue reduction time (MBRT). Continuous fermentation at 0.2 h(-1) dilution rate and sodium selenite addition gradient up to 0.69 g/L of Na(2)SeO(3) yielded 1. 89 g/L of biomass with 1904 microg of selenium/g of dry biomass. However, MBRT was 0.1 min, which indicated that the majority of selenium was in the inorganic form. On the other hand, continuous fermentation at 0.2 h(-1) dilution rate and sodium selenate gradient up to 0.28 g/L of Na(2)SeO(4) yielded 0.76 g/L of dry biomass with 687 microg of selenium/g of dry biomass, and MBRT was 26 min, which indicated a high concentration of organically bound selenium. Overall, the results indicate a Se/S ratio of 3.9:1 and a dry biomass/Se ratio of 5.5:1 as optimal for continuous production of organically bound selenium.     

A simulation model of the water uptake of a beech forest: testing variations in root biomass and distribution

A validated mathematical model was used to simulate the effects of roots biomass and distribution on the water uptake of a 120-year old beech (Fagus silvatica L.) stand located at Solling (West Germany). The modell was based on the soil-water movement equation including an extraction term to account for the water uptake of the root system. This extraction term varies with the meteorological conditions, root density and soil water potential. Root biomass values considered were: 500 kg. ha^?-1, 1000 kg. ha^?-1, 2000 kg. ha^?-1, 4000kg. ha^?-1 and 6000 kg. ha^?-1. For each of these values, five different distributions were tested. These ranged between a soil profile 0.40m deep with high root concentration to a 1.00m soil profile with idealized uniform root distribution. The model was run under two different rainfall conditions: i) an uncommon dry year (1971), (ii) a hypothetical normal year without a dry period. The model showed the fundamental importance of the root distribution under the given meteorological conditions. This was more evident when the root biomass was low and medium. A good root distribution throughout the profile allowed the forest to overcome long dry periods. With the maximum root biomass value the distribution had no effect on the forest water uptake.     

Production and carrying capacity for the earthworm Lumbricus terrestris in culture

Approximately 8 h were required at 25°C for food to pass from mouth to anus in the earthworm Lumbricus terrestris. Gut load per unit transit appeared inversely related to nitrogen content; values of about 2 and 44 mg dry castings per 100mg dry worm were obtained with activated sludge and mineral soil, respectively. Production of biomass was greater in a substrate of activated sludge and loam relative to activated sludge and cellulose, despite higher concentrations of nitrogen in the latter. Optimum population density was about 8 earthworms (31 g live wt) in 1000cm³ 2:1 sludge:soil. Growth occurred at a maximum rate between 15 and 25°C. A yield of approximately 4% biomass (dry wt) was obtained on a mixture of activated sludge and loam, based on the content of organic matter present.     

Effect of phosphorus supply on plant productivity,photosynthetic efficiency and bioactive-component production in Prunella vulgaris L. under hydroponic condition

We have examined the influence of four phosphorus (P) levels on growth performance, photosynthetic efficiency and bioactive phytochemical production of Prunella vulgaris L. in hydroponic culture. Results that 0.20 mM P was enhanced the dry weight, shoot height, spica and root weights, spica length and number, total chlorophyll and carotenoid contents, net photosynthetic rate, transpiration rate, and stomatal conductance were determined after three months' treatment. A supply of surplus P resulted in a higher concentration of foliar P and was negatively correlated with biomass. Both P-deficient (0 mM) and high P (10.00 mM) resulted in increased concentrations of ursolic acid and oleanolic acid, with the exception of flavonoids. An increase in water extract from P. vulgaris spicas was noted with the application of increasing P concentrations. Our results indicated that the application of 0.20 mM P improves the biomass production and the yield of bioactive constituents of P. vulgaris.     

Synthesis and antifungal activity of a series of N-substituted [2-(2,4-dichlorophenyl)-3-(1,2,4-triazol-1-yl)]propylamines

A series of N-mono- or N, N-disubstituted [2-(2,4-dichlorophenyl-3-(1,2,4-triazol-1-yl)]propylamines and N-[2-(2,4-dichlorophenyl-3-(1,2,4-triazol-1-yl)propyl]amides were synthesized and tested for their fungicidal activity in vitro and in vivo against a group of plant pathogenic fungi. Some compounds exhibited a fairly good in vitro activity. The replacement of the ether group of tetraconazole with a secondary or tertiary amino group leads to compounds that maintain the antifungal activity on several phytopathogenic fungi, provided that the substituents are not too bulky or lipophilic. The allyl, propargyl, and cyclopropyl groups appear particularly suitable. Although these compounds have some structural similarities with terbinafine and naftifine, which act as squalene epoxidase inhibitors, they maintain the usual mechanism of action of the other triazoles.     

Methyl jasmonate reduces chilling injury and maintains postharvest quality of mango fruit

Exposure of mango (Mangifera indica cv. Tommy Atkins) fruit to methyl jasmonate (MJ) vapors (10(-)(4) M) for 24 h at 25 degrees C reduced chilling injury during subsequent storage for 21 days at 7 degrees C and after 5 days of shelf life at 20 degrees C. The chilling tolerance induced by MJ was positively correlated with the reduction in the percent ion leakage of mango tissue. The overall quality of MJ-treated fruit was also better than that of control fruit. MJ treatment increased the total soluble solids but did not affect titratable acidity or pH. MJ also did not change the normal climacteric rise in respiration, water loss, and softening rates. The efficacy of MJ to reduce chilling injury and decay of mango could be related to the tolerance induced at low temperature. It was concluded that MJ treatment may prevent chilling injury symptoms of mango without altering the ripening process.     

Plasma Membrane Lipids Are the Powerful Components for Early Stage Aluminum Tolerance in Triticale

The order of aluminum (Al) tolerance in triticale lines (ST2>ST22) after re-elongation in an Al-free 0.2 mM calcium (Ca) solution for 9 h (Ca period) following 1 h pretreatment with 20 μM Al (Al period) agreed with that after 24 h of Al treatment. Permeability of the plasma membrane (PM) of root-tip cells after the Ca period was significantly increased in Al-sensitive ST22. Al was accumulated more heavily in the root-tip portion of ST22 than in that of ST2, although similar amounts of malic and citric acid anions were released from both triticale lines. We established a new system examining lipid permeability using synthesized nylon-2,8 ultrathin and porous capsules trapped previously with 0.1% (w/v) methylene blue solution and coated thereafter with PM lipid isolated from root tips by a newly developed technique. Permeability of the PM lipid measured with time in 0.2 mM Ca with or without 50 μM Al photometrically ( A ₆₈₀) was significantly greater in Al-sensitive ST22 after 5 min of Al treatment. This is the first report to directly show the primary and early role of PM lipid in Al tolerance in triticale.     

Zinc nutrition affects alfalfa responses to water stress and excessive moisture

The interactive effect of applied zinc (Zn) and soil moisture on early vegetative growth of three alfalfa (lucerne) (Medicago sativa L.) varieties was investigated in a sand‐culture pot experiment to test whether there is link between Zn nutrition and soil moisture stress or excessive moisture tolerance in alfalfa plants. Three varieties (Sceptre, Pioneer L 69, and Hunterfield) with differential Zn efficiency (ability of a variety to grow and yield well in a Zn deficient soil is called a Zn‐efficient variety) were grown at two Zn levels (low Zn supply: 0.05 mg Zn kg^‐1 of soil, adequate Zn supply: 2.0 mg Zn kg^‐1 of soil) and three levels of soil moisture (soil moisture stress: 3% soil moisture on soil dry weight basis; adequate soil moisture: 12% soil moisture on soil dry weight basis; excessive soil moisture: 18% soil moisture on soil dry weight basis) in a Zn deficient (DTPA Zn: 0.06 mg kg^‐1 soil) siliceous sand. Zinc treatments were applied at planting, while soil moisture treatments were applied three weeks after planting and continued for two weeks. Plants were grown in pots under controlled temperature conditions (20°C, 12 h day length; 15°C, 12 h night cycle) in a glasshouse. Plants grown at low Zn supply developed Zn deficiency symptoms, and there was a severe solute leakage from the leaves of Zn‐deficient plants. Adequate Zn supply significantly enhanced the leaf area, leaf to stem ratio, biomass production of shoots, and roots, succulence of plants and Zn concentration in leaves. At low Zn supply, soil moisture stress and excessive moisture treatments significantly depressed the shoot dry matter, leaf area and leaf to stem ratio of alfalfa plants, while there was little impact of soil moisture treatments when supplied Zn concentration was high. The detrimental effects of soil moisture stress and excessive soil moisture under low Zn supply were less pronounced in Sceptre, a Zn‐efficient alfalfa variety compared with Hunterfield, a Zn‐inefficient variety. Results suggest that the ability of alfalfa plants to cope with water stress and excessive soil moisture during early vegetative stage was enhanced with adequate Zn nutrition.     

Impact of cycloheximide addition on adenylates in soil

Cycloheximide inhibits specifically the ribosomal protein synthesis of eukaryotic cells, i.e. the metabolism of soil fungi. We measured cycloheximide effects on adenylates in 20 different soils (0-10 cm depth) from arable, grass and forest land with a large variety of soil properties. The aims were (1) to assess the interactions between cycloheximide effects and soil properties and (2) to prove the relationship between cycloheximide effects on ATP and the ergosterol-to-microbial biomass C ratio, which is an indicator for the fungal proportion of the total microbial biomass. The adenylates ATP, ADP and AMP were measured 6 h after adding either 10 mg cycloheximide per gram soil in combination with 24 mg talcum per gram soil or 24 mg talcum per gram soil solely. The medians of the relative increases in AMP and ADP were 45 and 25% and the medians of the relative decreases in ATP and adenylates were −36 and −12%. These changes in adenylate composition lead to a cycloheximide-induced relative decrease in the adenylate energy charge level of 15%. The relative decrease in ATP content after cycloheximide addition was significantly correlated with the ATP-to-microbial biomass C ratio, but not with the ergosterol-to-microbial biomass C ratio. The absolute increase in ADP and the absolute decrease in ATP were affected by the clay content according to principal component analysis. The reduction of the ATP-to-microbial biomass C ratio indicates that this ratio had the potential of being an important ecotoxicological indicator of direct toxic effects of organic pollutants on soil microorganisms.     

Hydrolysis of Chicoric and Caftaric Acids with Esterases and Lactobacillus johnsonii in Vitro and in a Gastrointestinal Model

Chicoric acid (ChA) and caftaric acid (CafA) were identified as bioactive components of chicory and have been ascribed a number of health benefits. This study investigated the hydrolysis of ChA and CafA with enzymes and a probiotic bacterium Lactobacillus johnsonii (La1). Esterase from Aspergillus japonicus (24 U/mg) hydrolyzed 100% of ChA (5 mM) and CafA (5 mM) after 3 h, at pH 7.0 and 37 °C. Under the same reaction conditions, 100% hydrolysis of ChA and CafA was achieved with a spray-dried preparation of La1. The addition of La1 (100 mg/mL, 3.3 E9 cfu/g) to CafA solution in a gastrointestinal model (GI model) resulted in 65% hydrolysis of CafA. This model simulates the physicochemical conditions of the human gastrointestinal tract. No hydrolysis of CafA was observed after passage through the GI model in the absence of La1. The results of this study support the hypothesis that ChA and CafA are degraded by gut microflora before absorption and metabolization.     

Effect of sample preparation,length of time,and sample size on quantification of total lipids from bovine liver

The objective was to evaluate the effect of sample preparation (pulverization under liquid nitrogen, homogenization, or sonication), time length of sonication (0-60 s), shaking in chloroform/methanol solvent (0, 2, 4, or 12 h), incubation in chloroform (0 or 12 h), and drying of extracted lipids at 50 degrees C (2, 4, 6, or 24 h), and sample size (50-250 mg) on quantification of total lipids from bovine liver. Pulverization under liquid nitrogen yielded the lowest recovery. Sonication was least time-consuming for sample preparation. Precise estimates and the greatest recovery were obtained with 30 s of sonication, at least 2 h of shaking in chloroform/methanol solvent, 12 h of incubation in chloroform, and at least 6 h of drying. Sample sizes of at least 150 mg gave precise estimates. The results demonstrate that sample preparation, time length of different steps of the extraction procedure, and sample size affect quantification of total lipid from bovine liver.     

Laboratory Assessment of Nostoc 9v (Cyanobacteria) Effects on N2 Fixation and Chemical Fertility of Degraded African Soils

The potential of Nostoc 9v for improving the nitrogen (N)₂–fixing capacity and nutrient status of semi‐arid soils from Tanzania, Zimbabwe, and South Africa was studied in a laboratory experiment. Nostoc 9v was inoculated on nonsterilized and sterilized soils. Inoculum rates were 2.5 mg dry biomass g^?1 soil and 5 mg dry biomass g^?1 soil. The soils were incubated for 3 months at 27 °C under 22 W m² illumination with a photoperiod of 16 h light and 8 h dark. The moisture was maintained at 60% of field capacity. In all soils, Nostoc 9v proliferated and colonized the soil surfaces very quickly and was tolerant to acidity and low nutrient availability. Cyanobacteria promoted soil N₂ fixation and had a pronounced effect on total soil organic carbon (SOC), which increased by 30–100%. Total N also increased, but the enrichment was, in most soils, comparatively lower than for carbon (C). Nitrate and ammonium concentrations, in contrast, decreased in all the soils studied. Increases in the concentration of available macronutrients were produced in most soils and treatments, ranging from 3 to 20 mg phosphorus (P) kg^?1 soil, from 5 to 58 mg potassium (K) kg^?1 soil, from 4 to 285 mg calcium (Ca) kg^?1, and from 12 to 90 mg magnesium (Mg) kg^?1 soil. Positive effects on the levels of available manganese (Mn) and zinc (Zn) were also observed.     

Pharmacokinetic profile of total quercetin after single oral dose of tartary buckwheat extracts in rats

As an important edible and medicinal material, tartary buckwheat ( Fagopyrum tataricum Gaertn.) is commonly used as a kind of food or drug in eastern Asian countries. To investigate the pharmacokinetic profile of total quercetin after a single oral dose of tartary buckwheat extract in rats, a sensitive, simple, and accurate HPLC-UV method was developed to determine total quercertin following plasma enzyme hydrolysis with glucuronidase/sulfatase. Eighteen male Wistar rats were randomly assigned into three groups, which were given three different doses of tartary buckwheat extract. The pharmacokinetic results showed that the area under the plasma concentration-time curve (AUC) of total quercetin after single oral doses of tartary buckwheat extract presented a linear relationship. The peak plasma concentrations (C(max)) of total quercetin afte plasma enzyme hydrolysis with glucuronidase/sulfatase were 0.55 ± 0.26, 1.10 ± 0.53, and 2.05 ± 0.26 μg/mL; the peak times (T(max)) were 2.33 ± 1.94, 2.75 ± 3.67, and 2.50 ± 1.82 h; the areas under the curves (AUC(0-36h)) were 5.29 ± 1.35, 10.02 ± 4.43, and 22.51 ± 3.05 μg·mL(-1)·h(-1) for three doses of tartary buckwheat extract (60, 120, and 240 mg/kg). The present study has provided a basic pharmacokinetic profile of total quercetin after a single oral dose of tartary buckwheat extract in rats.