Water-soluble Fraction Research Articles

Stable Isotope Dilution LC/ESI-SRM/MS Analysis for Highly Polar Bioactive Dipeptides Found in Fermented Brown Rice Product using a Porous Graphitic Carbon Column

In this study, a validated analytical method based on liquid chromatography/electrospray ionization-selected reaction monitoring/mass spectrometry (LC/ESI-SRM/MS) was developed for dipeptides in a functional food, fermented brown rice and rice bran with Aspergillus oryzae (FBRA). The aim of this study was to screen and quantify dipeptides in the water-soluble fraction of FBRA. There are few studies focusing on dipeptides in FBRA, even though FBRA is expected to contain bioactive peptides: FBRA or its aqueous extract has shown variety of biological activities. An anti-hypertensive dipeptide was found in similar rice bran products digested by thermolysin. Dipeptides are the smallest class of peptides, and many show a variety of biological activities. Some bioactive peptides are widely found in fermented foods. However, because dipeptides are generally too polar to be retained on versatile LC columns, such as octadecylsilyl columns, dipeptides have been often overlooked and not quantified because of the analytical difficulties. The LC was performed using a porous graphitic carbon column with the mobile phases of 0.1% formic acid/0.1% trifluoroacetic acid/1% tetrahydrofuran in water and acetonitrile. The stable isotope-labeled internal standards and SRM for dipeptides made it possible to develop a validated method. Five dipeptides were identified and quantified in FBRA in much higher concentrations than related materials (malted brown rice, rice bran, brown rice, and white rice) as follows: Ile-Arg, 82.1 µg/g; Ala-Phe, 27.8 µg/g; Ala-Tyr, 31.6 µg/g; Val-Phe, 46.3 µg/g; and Ile-Tyr, 49.9 µg/g. This method was simple and robust and would be applicable for other functional foods.Graphical

On the Influence of Wavelengths of Different Ranges of Photosynthetically Active Radiation on the Carbon Isotope Composition of Plant Biomass and Its Fractions (Using as the Example Lettuce (<i>Lactuca sativa</i> L.) of the Aficion Variety)

The influence of incident wavelengths of four ranges: short-wave red (623–641 nm), long-wave red (646–667 nm), far red (727–751 nm) and blue (452–477 nm) on the carbon isotopic composition of lettuce biomass (Lactica sativa L.) of the Aficion variety was studied. Lettuce was grown in climatic chambers, lighting was provided by irradiators based on narrow-band LEDs. Monochromatic blue and red light have the strongest multidirectional influence. Radiation from the blue range shifts the carbon isotopic composition of lettuce biomass towards enrichment in 12C. Radiation from the red range shifts the carbon isotopic composition of lettuce biomass towards enrichment in 13C. Based on the previously developed model of carbon isotope fractionation in a photosynthesizing cell, carbon isotopic shifts of plant leaf biomass were analyzed. It is shown that in the dark period the biomass is enriched in 12C. This is a consequence of dark respiration, during which the plant loses CO2 enriched in 13C. It is shown that the reason for the observed isotopic differences between the water-soluble and water-insoluble fractions of the leaf biomass is a result of the different participation of the assimilatory carbon flux, enriched in 12C, and the photorespiratory carbon flux, enriched in 13C, in the formation of these fractions during photosynthesis.

Effect of high-pressure homogenization treatment and Persian gum on rheological, functional properties and release kinetic of crocin from double emulsions

In the present study, the effect of water-soluble fraction of Persian gum (PG) and high-pressure homogenization (HPH) on some characteristics of double emulsions of (W/O/W) containing crocin was investigated. The results show the addition of PG increases the zeta potential and alters the charge on the droplet surface, thus reducing coalescence and improving emulsion stability. Also, the PG-coated emulsions have larger droplet sizes, but improved physical stability due to electrostatic and steric repulsions as well as high viscosity. HPH treatment leads to a decrease in droplet size and an increase in zeta potential, further improving emulsion stability. The findings suggest that the combination of PG and HPH treatment could be implemented to produce stable double emulsions for various applications like emulsions beverages containing crocin. Moreover, double emulsions stabilized with PG can positively affect the protection of crocin under gastrointestinal conditions, and more crocin can be absorbed by the body in the intestine. The dynamic behavior exhibited that the treated DEs using HPH behaved more like viscous fluids (G′< G″) when the frequency increased.

Maternal Obesity and Kawasaki Disease-like Vasculitis: A New Perspective on Cardiovascular Injury and Inflammatory Response in Offspring Male Mice.

Maternal obesity affects the risk of cardiovascular disease and inflammatory response in offspring. However, the impact of maternal obesity on offspring with Kawasaki disease (KD), the leading cause of childhood acquired heart disease, is still an understudied area. This study aimed to elucidate the impact of maternal obesity on offspring in KD-like vasculitis and the underlying mechanisms. Offspring of obese female mice and normal diet dams were randomly divided into two subgroups. The pups were injected intraperitoneally with either Candida albicans water-soluble fraction (CAWS) or phosphate buffered saline (PBS) to establish the obesity (OB)-CAWS group, OB group, wild type (WT)-CAWS group, and WT group. Their weight was monitored during the study. After four weeks, echocardiography was applied to obtain the alternation of cardiac structures. Mouse cytokine panel, Hematoxylin-Eosin (HE) staining, western blot, and real-time qPCR were used to study the pathological changes and protein and RNA expression alternations. Based on the study of pathology, serology and molecular biology, maternal obesity lead to more severe vasculitis and induced altered cardiac structure in the offspring mice and promoted the expression of pro-inflammatory cytokines through activating the NF-κB signaling pathway. Maternal obesity aggravated the inflammatory response of offspring mice in KD-like vasculitis.

Optimization of Enzyme-Assisted Extraction of Bioactive Compounds from Sea Buckthorn (Hippophae rhamnoides L.) Leaves: Evaluation of Mixed-Culture Fermentation.

Hippophae rhamnoides L. leaves possess a remarkable amount of polyphenols that could serve as a natural remedy in various applications. In comparison, numerous techniques, such as conventional and high-pressure techniques, are available for extracting the bioactive fractions from sea buckthorn leaves (SBL). However, enzyme-assisted extraction (EAE) of SBL has not been comprehensively studied. The aim of this study was to optimize critical EAE parameters of SBL using the cellulolytic enzyme complex, Viscozyme L, to obtain a high-yield extract with a high concentration of bioactive compounds. In order to determine the optimal conditions for EAE, the study employed a central composite design and response surface methodology to analyze the effects of four independent factors (pH, temperature, extraction time, and enzyme concentration) on two different responses. Our findings indicated that under optimal conditions (3:15 h extraction, temperature 45 °C, pH 4.9, and 1% Viscozyme L v/w of leaves DW), EAE yielded 28.90 g/100 g DW of the water-soluble fraction. Furthermore, the EAE-optimized liquid extract was continuously fermented using an ancient fermentation starter, Tibetan kefir grains, which possess lactic acid bacteria (LAB) and have significant potential for use in biopreservation. Interestingly, the results indicated various potential prebiotic characteristics of LAB. Additionally, alterations in the cell wall morphology of the SBL residue after EAE were examined using scanning electron microscopy (SEM). This study significantly optimized EAE parameters for sea buckthorn leaves, providing a promising natural source of bioactive compounds for various applications, such as nutraceuticals, functional foods, and high-value products.

Effect of Probiotic Microorganisms on Catalase Activity, Fractional Composition of Soluble Proteins, and Intestinal Microbiota of Honey Bee

Recently, there has been a trend toward the use of new effective natural preparations to fight diseases and improve the health of honey bees. It is also known that a well-balanced structure of the intestinal microbiota of honey bees is the basis for their growth, development, strengthening of the immune response, and resistance to infections. It has been established that some strains of lactic acid bacteria that have antibacterial, anti-inflammatory, and immunomodulatory properties, are promising for the development of broad-spectrum probiotic preparations based on them. Therefore, the aim of the work was to determine the effect of probiotic strains Lactobacillus сasei IMV B-7280 and L. plantarum IMV B-7679 on catalase activity, protein content and protein profile of hemolymph, as well as microbiota spectrum of different parts of the intestines of Apis mellifera honey bees. Methods. To conduct the research, a control and two experimental groups of 60-90 bees each were formed. The bees of the control group were fed 60% sugar syrup + 1 mL of distilled H2O for 28 days. The experimental group of bees D1 received 1 mL of 60% sugar syrup + 1 mL of aqueous suspension containing cells of the L. casei IMV B-7280 strain at a concentration of 1 ∙ 106 CFU/mL every day; experimental group of bees D2, in addition to 1 mL of 60% sugar syrup, received 1 mL of aqueous suspension containing cells of L. plantarum IMV B-7976 strain at a concentration of 1 ∙ 104 CFU/mL. Catalase activity of the whole organism tissues was determined using the ability of hydrogen peroxide to form a stable colored complex with molybdenum salts on a spectrophotometer at a wavelength of 410 nm against water. The amount of protein in the whole organism tissues was determined by the Lowry method. The content of total protein in the body of bees was carried out according to the Kjeldahl method. Determination of the content of individual fractions of soluble proteins of the hemolymph was carried out by the method of vertical electrophoresis in a 7.5% polyacrylamide gel. The relative content of protein fractions was determined using the TotalLab TL120 program and expressed as a percentage of the total pool. To determine the qualitative and quantitative spectrum of the gut microbiota of bees, the hindgut and midgut were sampled (separately) from bees of control and experimental groups. The obtained samples were plated on eight selective solid media for cultivation of different groups of microorganisms. Results. A tendency to increase the catalase activity of bee tissues after 28 days of L. casei IMV B-7280 strain use and a consistently higher activity of this enzyme throughout the experimental period under the action of L. plantarum IMV B-7679 strain was established. In the control group of 28th days, the content of bees and catalase activity remained at a constant level. It was shown that on the 14th day and total protein in the body of bees that received L. casei IMV B-7280 strain increased signifi cantly. Water-soluble fractions of hemolymph proteins were found in bees of both groups: γ-globulins, β-globulins, α2-globulins, and α1-globulins. It should be noted that the albumin fraction was not detected. It has been shown that the hindgut contains a much larger number of microorganisms than the midgut. The use of L. casei IMV B-7280 strain led to an increase in the number of lactic acid bacteria and bifidobacteria in both parts of the gut, as well as to a decrease in the number of staphylococci, streptococci, and microscopic fungi. The use of L. plantarum IMV B-7679 strain had a similar effect, but the changes in the composition of gut microbiome were less pronounced. Conclusions. The use of probiotic strains L. casei IMV B-7280 and L. plantarum IMV B-7679 for feeding bees under the conditions of a laboratory thermostat led to quantitative changes in the composition of the intestinal microbiota of bees, namely an increase in the number of lactic acid bacteria and bifidobacteria, as well as a decrease in the number of some other groups of microorganisms in the gut. Probiotic strains stimulated catalase activity of bee’s body tissues, increased the level of total protein, and did not significantly affect the ratio of hemolymph protein fractions.

Preparation and Characterization of Novel Green Seaweed Films from Ulva rigida.

Ulva rigida green seaweed is an abundant biomass consisting of polysaccharides and protein mixtures and a potential bioresource for bioplastic food packaging. This research prepared and characterized novel biodegradable films from Ulva rigida extracts. The water-soluble fraction of Ulva rigida was extracted and prepared into bioplastic films. 1H nuclear magnetic resonance indicated the presence of rhamnose, glucuronic and sulfate polysaccharides, while major amino acid components determined via high-performance liquid chromatography (HPLC) were aspartic acid, glutamic acid, alanine and glycine. Seaweed extracts were formulated with glycerol and triethyl citrate (20% and 30%) and prepared into films. Ulva rigida films showed non-homogeneous microstructures, as determined via scanning electron microscopy, due to immiscible crystalline component mixtures. X-ray diffraction also indicated modified crystalline morphology due to different plasticizers, while infrared spectra suggested interaction between plasticizers and Ulva rigida polymers via hydrogen bonding. The addition of glycerol decreased the glass transition temperature of the films from -36 °C for control films to -62 °C for films with 30% glycerol, indicating better plasticization. Water vapor and oxygen permeability were retained at up to 20% plasticizer content, and further addition of plasticizers increased the water permeability up to 6.5 g·mm/m2·day·KPa, while oxygen permeability decreased below 20 mL·mm/m2·day·atm when blending plasticizers at 30%. Adding glycerol efficiently improved tensile stress and strain by up to 4- and 3-fold, respectively. Glycerol-plasticized Ulva rigida extract films were produced as novel bio-based materials that supported sustainable food packaging.

Effect of preserved eggs on the health of SD rats, and anti-tumor action of HT-29 cells.

Preserved eggs are traditional alkali-pickled food in China and have been enjoyed by consumers and extensively studied by researchers for their nutritional tastes and their anti-tumor, anti-inflammatory, antioxidant, lipid-lowering, and blood pressure-lowering properties. To study the anti-tumor effects of preserved eggs, this project observed the health on rats, and anti-tumor effects and separated anti-tumor active components on HT-29 cells. SD rats fed for 80 days showed that preserved eggs had no significant effect on weight, food intake, blood pH, liver tissues, or organ indices. Preserved eggs significantly increased blood levels of oxidative stress markers SOD and CAT, decreased MDA levels by 0.46, 0.23, and 0.25 times. Moreover, they also increased the level of IL-2 from 1233 to 1340 pg/mL. Two water-soluble bioactive peptide fractions, B1 and B2, with molecular weights ≥10 kDa were further obtained from preserved eggs by ultrafiltration and Superdex Peptide 10/300 GL. The potential mechanism of B1 and B2 is to activate the internal mitochondrial apoptotic pathway and induce apoptosis by up-regulating the expression of the pro-apoptotic factors cytochrome C, caspase-3, and caspase-9 mRNA in HT-29 cells.

Exploring the antioxidant and antimicrobial properties of the water-soluble fraction derived from pyrolytic lignin separation in fast-pyrolysis bio-oil

Exploring the antioxidant and antimicrobial properties of the water-soluble fraction derived from pyrolytic lignin separation in fast-pyrolysis bio-oil

GROWTH PERFORMANCE AND SELECTED BIOCHEMICAL INDICES ASSESSMENT IN THE SERUM AND LIVER OF CLARIAS GARIEPINUS JUVENILES EXPOSED TO VARYING CONCENTRATIONS OF WATER SOLUBLE FRACTION OF CRUDE OIL

The effect of varying concentrations (0, 0.25, 0.05 and 1.0% v/v) of water soluble fraction (WSF) of crude oil on the growth performance and biochemical changes in the serum and liver of 144 Clarias gariepinus Juveniles stocked into 12 glass aquaria at the rate of 12 fish per tank in triplicate for 90 days was assessed. The growth performance indices of the fish juvenile over the 90 days of toxic exposure revealed that the mean weight gained, specific growth rate, and total feed intake of the fish specimen decreases with increase concentration of the WSF of crude oil. However, 10% and 20% mortality were recorded in the fish exposed to 0.5% and 1.0% WSF of crude oil respectively. The activity of ALT and AST in the serum of the fish were significantly higher (p&lt;0.05) with the increased concentration of WSF of crude oil while the ALT and AST activity of the liver of the fish juveniles irrespective of the added concentration of WSF of crude oil showed no significant differences (p&gt;0.05). The study concluded that WSF of crude oil had adverse effects on the growth performance, blood serum and liver activities in C. gariepinus juveniles.

Apoptotic volume decrease (AVD) in A549 cells exposed to water-soluble fraction of particulate matter (PM10).

Exposure to atmospheric particulate matter (PM) is recognized as a human health risk factor of great concern. The present work aimed to study the cellular mechanisms underlying cytotoxic effects of airborne particulate matter <10µm in size (PM10), sampled in an urban background site from January to May 2020, on A549 cells. In particular, the study addressed if PM10 exposure can be a main factor in the induction of the Apoptotic Volume Decrease (AVD), which is one of the first events of apoptosis, and if the generation of intracellular oxidative stress can be involved in the PM10 induction of apoptosis in A549 cells. The cytotoxicity of PM10 samples was measured by MTT test on cells exposed for 24h to the PM10 aqueous extracts, cell volume changes were monitored by morphometric analysis of the cells, apoptosis appearance was detected by annexin V and the induction of intracellular oxidative stress was evaluated by the ROS sensitive CM-H2DCFDA fluorescent probe. The results showed cytotoxic effects ascribable to apoptotic death in A549 cells exposed for 24h to aqueous extracts of airborne winter PM10 samples characterized by high PM10 value and organic carbon content. The detected reduced cell viability in winter samples ranged from 55% to 100%. Normotonic cell volume reduction (ranging from about 60% to 30% cell volume decrease) after PM10 exposure was already detectable after the first 30min clearly indicating the ability of PM10, mainly arising from biomass burning, to induce Apoptotic Volume Decrease (AVD) in A549 cells. AVD was prevented by the pre-treatment with 0.5mM SITS indicating the activation of Cl- efflux presumably through the activation of VRAC channels. The exposure of A549 cells to PM10 aqueous extracts was able to induce intracellular oxidative stress detected by using the ROS-sensitive probe CM-H2DCFDA. The PM10-induced oxidative stress was statistically significantly correlated with cell viability inhibition and with apoptotic cell shrinkage. It was already evident after 15min exposure representing one of the first cellular effects caused by PM exposure. This result suggests the role of oxidative stress in the PM10 induction of AVD as one of the first steps in cytotoxicity.

Characteristics of Organic Matter Composition and Oxidation Potential in Road Dust in Winter in Xi'an

Roads are the main places where urban people are exposed to atmospheric particulate matter from outdoor activities, and certain oxidatively active substances contained in road particulate matter are important components that induce the generation of reactive oxygen species (ROS), which in turn endanger human health. Here, we explored the characteristics of organic matter composition in water-soluble (WSM) and methanol-soluble fractions (MSM) of road dust in Xi'an and its oxidation potential (OP). Additionally, we investigated the organic fractions and their distribution based on parallel factor analysis (PARAFAC) and analyzed the correlation between organic matter types and OP. The results showed that the water-insoluble fraction of road dust in Xi'an contained more chromophoric organic matter with an average total concentration of (4.71±1.27)×104 R.U., which was 12 times higher than that of WSM[(3.96±1.10)×103 R.U.], of which low-oxidizing humic-like substances (HULIS) were the main organic matter (34.8%-43.7% of the total organic matter). The results of cluster analysis showed that the important sources of organic matter in road dust in Xi'an were fuel combustion and industrial production. The mean value of dust oxidative toxicity was (0.34±0.08) pmol·(min·μg)-1, with the water-insoluble fraction providing 70% of the total oxidative toxicity of dust particles, which was 2.4 times higher than the water-soluble fraction. The main precursors of oxidative toxicity of dust particles were metal elements, and special types of organic substances were also one of the important oxidative toxicity precursors, among which chromophore organic matter was the main cause of OP production in the WSM fraction (r=0.35, P<0.01), and protein-like organic matter and highly oxidized HULIS in WSM may have been the main two types of organic substances for OP production. However, there was no significant correlation between organic matter concentration in MSM and water-insoluble OP (OPTotal-OPWSM) (r=-0.04, P>0.1), so the oxidative toxicity of the water-insoluble particulate matter fraction was mainly generated from non-organic matter.

Spawning time in adult polar cod (Boreogadus saida) altered by crude oil exposure, independent of food availability

ABSTRACT Fish early life stages are well known for their sensitivity to crude oil exposure. However, the effect of crude oil exposure on adults and their gametes during their spawning period is not well studied. Polar cod, a key arctic fish, may be at risk for crude oil exposure during this potentially sensitive life stage. Additionally, this species experiences lower food availability during their spawning season, with unknown combined consequences. In the present study, wild-caught polar cod were exposed to decreasing levels of a water-soluble fraction (WSF) of crude oil or control conditions and fed either at a low or high feed ration to assess the combined effect of both stressors. Samples were taken during late gonadal development, during active spawning (spawning window), and in the post-spawning period. Histology analysis of gonads from fish sampled during the spawning window showed that oil-exposed polar cod were more likely to have spawned compared to controls. Oil-exposed females had 947 differentially regulated hepatic genes, and their eggs had a higher polycyclic aromatic hydrocarbon body burden compared to controls. Feed ration did not consistently affect polar cod’s response to oil exposure for the endpoints measured, however, did alone result in decreases in some sperm motility parameters. These results suggest that polar cod’s spawning period is a sensitive life event to crude oil exposure, while feed limitation may play a minor role for this supposedly capital breeder. The effects of adult exposure to crude oil on gamete quality and the next generation warrant further investigation.

Novel Insight into the Photophysical Properties and 2D Supramolecular Organization of Poly(3,4-ethylenedioxythiophene)/Permodified Cyclodextrins Polyrotaxanes at the Air-Water Interface.

Two poly(3,4-ethylenedioxythiophene) polyrotaxanes (PEDOT∙TMe-βCD and PEDOT∙TMe-γCD) end-capped by pyrene (Py) were synthesized by oxidative polymerization of EDOT encapsulated into TMe-βCD or TMe-γCD cavities with iron (III) chloride (FeCl3) in water and chemically characterized. The effect of TMe-βCD or TMe-γCD encapsulation of PEDOT backbones on the molecular weight, thermal stability, and solubility were investigated in depth. UV-vis absorption, fluorescence (FL), phosphorescence (PH), quantum efficiencies, and lifetimes in water and acetonitrile were also explored, together with their surface morphology and electrical properties. Furthermore, dynamic light scattering was used to study the hydrodynamic diameter (DH) and z-potential (ZP-ζ) of the water soluble fractions of PEDOT∙TMe-βCD and PEDOT∙TMe-γCD. PEDOT∙TMe-βCD and PEDOT∙TMe-γCD exhibited a sharp monodisperse peak with a DH of 55 ± 15 nm and 122 ± 32 nm, respectively. The ZP-ζ value decreased from -31.23 mV for PEDOT∙TMe-βCD to -20.38 mV for PEDOT∙TMe-γCD, indicating that a negatively charged layer covers their surfaces. Surface pressure-area isotherms and Brewster angle microscopy (BAM) studies revealed the capability of the investigated compounds to organize into sizeable and homogeneous 2D supramolecular assemblies at the air-water interface. The control of the 2D monolayer organization through the thermodynamic parameters of PEDOT∙TMe-βCD and PEDOT∙TMe-γCD suggests potential for a wide range of optoelectronic applications.

Anthrone method combined with adsorption of interferents as a new approach towards reliable quantification of total carbohydrate content in pyrolysis bio-oils

Carbohydrates are a group of compounds abundantly present in pyrolysis bio-oils. Their quantitative analysis however presents many challenges such as possible co-elution with other oxygenates when liquid chromatographic methods are used. For a cheaper and faster analysis, spectrophotometric methods are a convenient alternative. Most of the absorption spectrometry-based methods stem from the reaction scheme, where the carbohydrates are converted into furfural or 5-hydroxymethylfurfural (HMF) derivatives and simultaneously react with colorizing agents such as phenol or anthrone. Alternatively, the produced compounds can be determined in the UV region without the addition of colorizing agent. An obvious problem arises when these furanic compounds are already present in the mixture, which is the case with most biomass-based pyrolysis bio-oils. In this study, we quantified the responses of 26 potential interferents that are expected to be present in the water-soluble fraction of bio-oils as well as 13 different carbohydrates. To mitigate the interferences, we used styrene-divinylbenzene polymeric resin adsorbents to separate the less polar interferents from the aqueous bio-oil extract and naturally formed bio-oil aqueous phases. A derivatization-GC-MS method was used as a reference for comparison of the results. We found that the adsorption step is crucial for acquiring more precise and accurate results. The anthrone method was the least susceptible to interferences and simultaneously produced results that were the closest to the reference GC-MS method. Statistical analysis showed no significant differences between the optimized anthrone method and the reference GC-MS method.

Identification of Anti-Inflammatory Compounds from Peucedanum praeruptorum Roots by Using Nitric Oxide-Producing Rat Hepatocytes Stimulated by Interleukin 1β.

The roots of Peucedanum praeruptorum Dunn and Angelica decursiva Franchet et Savatier are designated Zenko, which is a crude drug defined by the Japanese Pharmacopoeia. This crude drug is used as an antitussive and an expectorant and is included in the Kampo formula Jinsoin, which improves cough, fever, and headache. Although the anti-inflammatory effects of this crude drug have been determined, the constituents responsible for this effect remain unknown. To investigate biologically active compounds, rat hepatocytes were used, which produce proinflammatory mediator nitric oxide (NO) in response to proinflammatory cytokine interleukin 1β (IL-1β). A methanol extract of P. praeruptorum roots, which suppressed IL-1β-induced NO production, was fractionated into three crude fractions (ethyl acetate (EtOAc)-soluble, n-butanol-soluble, and water-soluble fractions) based on hydrophobicity. The EtOAc-soluble fraction markedly inhibited NO production. After this fraction was purified, three biologically active compounds were identified as praeruptorins A, B, and E, the contents of which were high. A comparison of their activities indicated that praeruptorin B exhibited the highest potency to inhibit NO production by decreasing inducible NO synthase expression and suppressed the expression of mRNAs encoding proinflammatory cytokines. Collectively, the three praeruptorins may primarily contribute to the anti-inflammatory effects of P. praeruptorum roots.

Effects of dissolved organic matter on mercury speciation in rice rhizosphere amended with sulfur-rich biochar

Natural sulfur (S)-rich biochar (NRB) can be employed as an alternative for traditional S-modified biochar. However, the effect of dissolved organic matter (DOM) on mercury (Hg) speciation in rice rhizosphere soils under natural S-rich biochar application remains unclear. We conducted a pot experiment to study the effects of NRB application on the chemical composition and structure of DOM and the related speciation and availability of Hg in rice rhizosphere. Applying NRB significantly increased the concentration of methylmercury (MeHg) in the rhizosphere soils, which was enhanced with application frequency. This observation can be explained by MeHg immobilization in response to increasing S content in rice rhizosphere soils. We also observed increased molecular weight and functional group complexity of DOM, likely contributing to the decrease in MeHg mobility. Furthermore, the increase in pH and humification of DOM caused by S-rich biochar application generally reduced the concentrations of water-soluble and mercuric-sulfide fraction (easily-available Hg species) and organo-chelated fraction (potentially-available Hg species). Our findings highlight that the application of NRB can reduce the availability of MeHg in rice rhizosphere, thus providing a practical basis for reducing the potential risk of MeHg toxicity.

Screening for In Vitro Antibacterial Activity of Leaves Extracts of Certain Selected Plants against Rhodococcus equi

In the present study, the research work was carried out during June 2018 to March 2019 at ICAR-NRCE-EPC (Indian Council of Agricultural Research, National Research Centre on Equines, Equine Production Campus), Jorbeer, Bikaner (Rajasthan), India. The study was conducted to investigate in vitro antibacterial activity of ethanolic, chloroformic and Sequentially Extracted Water Extract (SEWE) leaves extracts of Aegle marmelos, Morus alba, Nerium indicum, Cascabela thevetia (L.) Lippold, Plumeria alba L. and Azadirachta indica against Vap A and Vap C positive Rhodococcus equi. Fresh leaves of these plants were collected manually from campus of ICAR-NRCE-EPC, Jorbeer, Bikaner (Rajasthan). In initial screening ethanolic leaves extract of these plants except Azadirachta indica, were found non-active against Rhodococcus equi. Chloroformic leaves extracts of Azadirachta indica did not showed in vitro antibacterial activity against Rhodococcus equi. While ethanolic and Sequentially Extracted Water Extract (SEWE) leaves extracts of Azadirachta indica showed good in vitro antibacterial activity against Rhodococcus equi. Further, solvent based fractionation, Ethanol Soluble Fraction (ESF), Methanol Soluble Fraction (MSF) and Water Soluble Fraction (WSF) of polar compounds of SEWE did not showed in vitro antibacterial activity against Rhodococcus equi. On comparison with currently used antibiotics (azithromycin and rifampicin), required concentration of the leaves extract of Azadirachta indica was too high for their possibilities of in vivo use, so abundant availability of Azadirachta indica leaves and their activity against Rhodococcus equi suggests their potential for use as disinfectant against Rhodococcus equi.

Analysis and subcellular distribution of organophosphate esters (OPEs) in rice tissues.

Recent studies have identified the ability of plants to uptake and translocate organophosphate esters (OPEs) within cells. In response to the increasing interest in OPEs and their occurrence in paddy fields and rice, the current study aimed to present an effective and sensitive GC-MS based methodology for quantitative determination of 11 OPEs with octanol-water coefficients ranging from 1.6 to 10. Rice was sonicated with hexane and dichloromethane, and fractionated on two columns: one consisting of neutral alumina, andneutral silica, and the other consisting of graphitized carbon black. Method precision was validated using spiked rice (n = 30) and procedural blanks (n = 9). The mean recovery of matrix spikes for all target OPEs were within 78-110% with relative standard deviation lower than 25%, with a few exceptions. This method was applied to process the wild rice (O. sativa) in which tri-n-propyl phosphate was the dominant targeted OPE. The recoveries of surrogate standards were 81 ± 17% for d12- tris(2-chloroethyl) phosphate and 95 ± 8.8% for 13C12- triphenyl phosphate. The developed method was further used to examine the recoveries of target OPEs in the subcellular structure of rice tissues, including cell wall, cell organelles, cell water-soluble fractions, and cell residue. Recoveries of most target OPEs were in the range of 50-150%; however, ion enhancement was observed for four OPEs in root and shoot tissues. Hydrophobic OPEs accumulated in the cell wall, cell residue, and cell organelles while chlorinated OPEs mainly distributed in the cell water-soluble fraction. These results provide new insight for the ecological risk assessment of OPEs in an important food staple.

ZALEZNO ˙ S´ C MI ˛EDZY WYBRANYMI CECHAMI JAKO ´ SCIOWYMI WIEPRZOWEGO MI ˛E ´ SNIA ´ LONGISSIMUS A ZAWARTOSCI ˛A W NIM BIAŁEK ROZPUSZCZALNYCH W WODZIE

This study comprised 136 meat samples (m. longissimus lumborum) from 136 pork carcasses of six-month-old crossbred porkers derived from ♀ (Polish Large White×Polish Landrace)×♂ (Duroc×Pietrain), with an average carcass weight of 85.8 ± 8.5 kg, slaughtered on an industrial processing line. The pH1, pH24, pH48, WHC, colour parameters (L*, a*, b*), moisture content, crude protein, intramuscular fat, and water-soluble protein content were determined. Significantly high correlation coefficients between the proportion of water-soluble proteins in meat and crude protein and pH48, WHC, and lightness (L*) were recorded, with pH48 (measured in water extracts of meat) having the greatest effect on the proportion of water-soluble protein fractions in the meat and crude protein. In contrast, the basic chemical composition of the meat, i.e. moisture, crude protein, and intramuscular fat content, had no significant effect on the levels of water-soluble proteins. The results indicate that water-soluble proteins are a useful indicator of the quality of pork longissimus muscle.