Chloroform-methanol Mixture Research Articles

A Preliminary Antidepressant Assessment of a Novel Piperoyl Amide (Y3) Isolated and Elucidated from Piper nigrum Drupes Extract, Justifying Interaction Over MAO-A enzyme: An in vivo, in vitro and Molecular Docking Approach

This research aims to confirm the preliminary antidepressant activity of a newly discovered piperoyl amide (Y3) obtained from the extract of Piper nigrum (P. nigrum) drupes, through the inhibition of MAO-A enzyme. Column chromatography was employed for isolation and purification of Novel piperoyl-alkyl amide (Y3) which was eluted in a solvent mixture of Chloroform-Methanol (97:3). Structure elucidation was performed by implying spectroscopic techniques. Invitro in-comparison MAO activity assessment was performed between Y3 and piperine further confirmed by Molecular docking via Auto Dock Tools 1.5.6, Vina, and PyMOL. Invivo antidepressant activity was evaluated by the Despair Test in albino mice. Novel piperoyl-alkyl amide (Y3) was isolated and purified by column chromatography and elucidated spectroscopically. The antidepressant activity of Y3 at 10 and 20 mg/kg doses shown a significant decrease in immobility-time (IT) in contrast to stressed-control group at 107.0±7.6 sec (P<0.05) and 96.0±7.6 sec (P<0.01), respectively. Y3 shown a decent MAO-A inhibitory activity but lower than piperine showing IC50=20.24 µM, Vmax=1.375±0.3 nM/min/mg, Km=1.379×10-16 µM with a stronger interaction and better affinity (-12.6 kcal/mol) towards the active-site of MAO-A. Y3 was isolated and elucidated as (12E,14E)-15-(benzo[d] [1,3] dioxol-5-yl)-N-ethylpentadeca-12,14-dienamide. It proved a good antidepressant at 10 and 20 mg/kg doses. Molecular docking over MAO-A and QSAR studies depicted a better affinity score of Y3 than Piperine. Keywords: Piperoyl amide, Structure elucidation, antidepressant activity, molecular modeling, Piperine

Effect of the extraction method on the lipid composition of purslane (Portulaca oleracea L.) seed oil

Purslane (Portulaca oleracea L.) is a widespread weed plant used since ancient times as remedy and as food. Its seed oil possesses significant bioactive potential due to the high content of important phytonutrients, mainly essential fatty acids and phytosterols. Although the total fatty acid composition of oil is well documented, till now there is no data published about individual lipid classes. The information on sterols is scarce as well. Therefore, the aim of this work was to characterize in details for the first time the individual lipid classes and their fatty acid composition, sterols, as well as the oxidative stability of the oil, depending on the extraction method by either hexane, chloroform–methanol mixtures or super-critical CO2. The results revealed no significant effect of extraction method on the individual lipid classes (monoacylglycerols, diacylglycerols, free fatty acids, free sterols, triacylglycerols, sterol esters, wax esters and hydrocarbons), including the fatty acid composition of saponifiables, and quite weak effect on the oxidative stability of the oil regarding super-critical CO2 vs. organic solvents. Detailed analyses of lipid composition confirmed the potential of purslane seed oil as a cheap and highly valuable source of phytonutrients as essential fatty acids and phytosterols, for application in food, pharmaceutical and cosmetic industries.

CuI-catalyzed ligand-free synthesis of 2-aryloxazolo[5,4-b]-5,10,15,20-tetraarylporphyrins

A convenient ligand-free synthetic procedure has been developed to synthesize novel nickel(II) and copper(II) 2-aryloxazolo[5,4-b]-5,10,15,20-tetraarylporphyrins in 60–72% yields by a CuI-catalyzed intramolecular debrominative cyclization of 2-aroylamino-3-bromo-5,10,15,20-tetraarylporphyrins in dimethylsulfoxide containing potassium carbonate as a base. Subsequently, the copper(II) 2-(4-iodophenyl)oxazolo[5,4-b]-5,10,15,20-tetraphenylporphyrin underwent demetallation in the presence of concentrated sulfuric acid and afforded 2-(4-iodophenyl)oxazolo[5,4-b]-5,10,15,20-tetraphenylporphyrin in 75% yield which on zinc insertion using zinc(II) acetate in chloroform-methanol mixture provided zinc(II) 2-(4-iodophenyl)oxazolo[5,4-b]-5,10,15,20-tetraphenylporphyrin in 78% yield. The structures of newly constructed 2-aryloxazolo[5,4-b]-5,10,15,20-tetraarylporphyrins were assigned on the basis of spectral data and single crystal X-ray analysis. On photophysical investigation, these porphyrin analogues exhibit similar electronic properties as reported in the case of β-imidazole/thiazole-fused meso-tetraarylporphyrins.

Antioxidant potentials of five flavonoids compounds isolated from Varthemia iphionoids

Purpose: To evaluate the in vitro antioxidant activity of some flavonoids isolated from Varthemia iphionoids aqueous methanol extract.Methods: A portion (330 g) of the aqueous methanol extract of Varthemia iphionoids was adsorbed onto 200 g of S silica gel (70 - 230 mesh) and then loaded onto a column of S silica gel (70 - 230 mesh, 100 g). The column was packed with chloroform and eluted with chloroform-methanol mixtures of increasing polarity. Column chromatography purification (silica gel, 400 mesh) and thin-layer chromatography (TLC) or recrystallization yielded 5 compounds subsequently identified by nuclear magnetic resonance (NMR) and mass spectroscopy (MS).Results: The antioxidant activities of five identified flavonoids: Kumatakillin (1), Penduletin (2), Jeceidine (3), 6-Methoxy isokaemfride (4), and 3,3’-Di-O-methyl quercetin (5) were determined. The results indicated that compound 3 exhibited the best scavenging activity against DPPH radical and H2O2 comparable to that of quercetin as standard. All compounds had the ability to prevent the oxidation of β- carotene. In anti-hemolytic assay, the five compounds gave HT50 higher than vitamin C. The best protection activity of erythrocytes membrane was recorded with compound 2 (HT50 = 60.14 ± 0.72 min). Moreover, all compounds had significantly increased blood clotting time, and among them, compounds 1, 2, and 3 exhibited significantly increased thrombolytic activity compared to the control (***p < 0.001).Conclusion: These results indicate that Varthemia iphionoids has good antioxidant activities but further studies on the plant extract are required.

Conservation of freshwater eels in food‐web studies: Non‐lethal stable isotope analyses substitute fin for muscle tissue with lipid correction

AbstractAquatic food‐web studies involving fish and using stable isotope analysis typically sample fish white muscle, which generally requires the fish to be euthanised. Alternatively, fin tissue can be sampled non‐lethally, avoiding the need to euthanise an animal to obtain what is usually a relatively small tissue sample. We set out to develop equations for converting δ13C and δ15N values from derived non‐lethally sampled fin tissue to equivalent white muscle tissue, for use in food‐web studies. We explored δ13C and δ15N values of paired fin‐clips and white muscle from two species of anguillid fishes, the endemic longfin eel (Anguilla dieffenbachii), and the Australasian shortfin eel (A. australis), sourced from multiple studies spread throughout New Zealand. We also examined the effect of lipid extraction on δ13C and δ15N, which reduced variability of δ13C when compared to untreated samples. We were also able to estimate δ13C values of equivalent lipid‐extracted muscle samples directly from untreated fin. For δ15N, paired untreated fin and muscle values were not statistically different so fin δ15N can be used as a direct estimate of muscle δ15N. Solvent lipid extraction with a methanol–chloroform mixture, however, altered δ15N values and increased variability, so should be avoided. We present equations to estimate lipid‐corrected δ13C values of muscle tissue for use in stable isotope studies of freshwater anguillids, avoiding euthanasia. Our results demonstrate that δ13C and δ15N values can be accurately derived from non‐lethally obtained fin clips, reducing unnecessary mortality in fish species of conservation concern.

Influence of progressive halogenation of Zn(II)-tetraarylporphyrins and their free bases on the structure and spectral-fluorescence properties of tetrapyrrolic macrocycle

Zn(II)-2,3,7,8,12,13,17,18-octabromo-5,10,15,20-tetra-(2,6-difluorophenyl)porphyrin was synthesized by the interaction of Zn(II)-5,10,15,20-tetra-(2,6-difluorophenyl)porphyrin with N-bromosuccinimide in a mixture of chloroform-methanol, chloroform-dimethylformamide and in dimethylformamide. Zn(II)-2,3,7,8,12,13,17,18-octachloro-5,10,15,20-tetra-(2,6-difluorophenyl)porphyrin was formed by the reaction of 2,6-difluoro-substituted Zn(II)-porphyrin with N-chlorosuccinimide in the chloroform-methanol mixture and dimethylformamide. Zn(II)-2,3,7,8,12,13,17,18-octachloro-5,10,15,20-tetra-(2,3,4,5,6-pentafluorophenyl) porphyrin was synthesized from Zn(II)-5,10,15,20-tetra-(2,3,4,5,6-pentafluorophenyl)porphyrin with an excess of N-chlorosuccinimide via chlorination in dimethylformamide. The corresponding porphyrin-ligands were obtained by treating halogen-substituted zinc porphyrins with trifluoroacetic acid. The synthesized compounds were characterized by UV–Vis, fluorescence, 1H NMR spectroscopy and mass spectrometry methods. The structures of halogen-substituted porphyrins and their Zn(II) complexes were calculated by the DFT method. The effect of the β-pyrrole halogenation on fluorescence quantum yields of the synthesized compounds was estimated. These halogenated products can be employed for covalent functionalization of the macrocycle pyrrole fragments with the aim of imparting practically important optical, electrochemical and red-ox properties to them.

P-037. Revisiting ‘3 delays model[StQuote] for critical analysis of hypertensive maternal mortality in a developing nation

Zn(II)-2,3,7,8,12,13,17,18-octabromo-5,10,15,20-tetra-(2,6-difluorophenyl)porphyrin was synthesized by the interaction of Zn(II)-5,10,15,20-tetra-(2,6-difluorophenyl)porphyrin with N-bromosuccinimide in a mixture of chloroform-methanol, chloroform-dimethylformamide and in dimethylformamide. Zn(II)-2,3,7,8,12,13,17,18-octachloro-5,10,15,20-tetra-(2,6-difluorophenyl)porphyrin was formed by the reaction of 2,6-difluoro-substituted Zn(II)-porphyrin with N-chlorosuccinimide in the chloroform-methanol mixture and dimethylformamide. Zn(II)-2,3,7,8,12,13,17,18-octachloro-5,10,15,20-tetra-(2,3,4,5,6-pentafluorophenyl) porphyrin was synthesized from Zn(II)-5,10,15,20-tetra-(2,3,4,5,6-pentafluorophenyl)porphyrin with an excess of N-chlorosuccinimide via chlorination in dimethylformamide. The corresponding porphyrin-ligands were obtained by treating halogen-substituted zinc porphyrins with trifluoroacetic acid. The synthesized compounds were characterized by UV–Vis, fluorescence, 1H NMR spectroscopy and mass spectrometry methods. The structures of halogen-substituted porphyrins and their Zn(II) complexes were calculated by the DFT method. The effect of the β-pyrrole halogenation on fluorescence quantum yields of the synthesized compounds was estimated. These halogenated products can be employed for covalent functionalization of the macrocycle pyrrole fragments with the aim of imparting practically important optical, electrochemical and red-ox properties to them.

P-005. Involvement of RAGE pathway in visceral adipose tissue in preeclampsia

Zn(II)-2,3,7,8,12,13,17,18-octabromo-5,10,15,20-tetra-(2,6-difluorophenyl)porphyrin was synthesized by the interaction of Zn(II)-5,10,15,20-tetra-(2,6-difluorophenyl)porphyrin with N-bromosuccinimide in a mixture of chloroform-methanol, chloroform-dimethylformamide and in dimethylformamide. Zn(II)-2,3,7,8,12,13,17,18-octachloro-5,10,15,20-tetra-(2,6-difluorophenyl)porphyrin was formed by the reaction of 2,6-difluoro-substituted Zn(II)-porphyrin with N-chlorosuccinimide in the chloroform-methanol mixture and dimethylformamide. Zn(II)-2,3,7,8,12,13,17,18-octachloro-5,10,15,20-tetra-(2,3,4,5,6-pentafluorophenyl) porphyrin was synthesized from Zn(II)-5,10,15,20-tetra-(2,3,4,5,6-pentafluorophenyl)porphyrin with an excess of N-chlorosuccinimide via chlorination in dimethylformamide. The corresponding porphyrin-ligands were obtained by treating halogen-substituted zinc porphyrins with trifluoroacetic acid. The synthesized compounds were characterized by UV–Vis, fluorescence, 1H NMR spectroscopy and mass spectrometry methods. The structures of halogen-substituted porphyrins and their Zn(II) complexes were calculated by the DFT method. The effect of the β-pyrrole halogenation on fluorescence quantum yields of the synthesized compounds was estimated. These halogenated products can be employed for covalent functionalization of the macrocycle pyrrole fragments with the aim of imparting practically important optical, electrochemical and red-ox properties to them.

Allomones confer resistance to Musa cultivar Pisanglilin against infestation by Odoiporus longicollis [Oliver] and characterization of Allomones

In bioassay guided extraction of pseudostem powder of Pisanglilin by organic solvents we found the larvicidal activity in acetone extract, whose column chromatography by methanol-chloroform mixture separated the extract into 9-fractions, of which the 8th fraction showed larvicidal activity. Subfractionation of the active fraction by column chromatography resulted in the isolation of two larvicidal molecules [Stigmasterol-3-O-glucoside (SOG) and Sulfoquinovosyl diacylglycerol (SQDG)]. Yield of SOG was 0.002 % and SQDG was 0.005 % and both were highly toxic to O. longicollis larvae with LD50 of 0.40 and 0.378 ppm, respectively. Larvae fed these compounds stopped feeding on third day and died within one week. SOG inhibited the amylase and protease activity of gut and induced histolysis in the mid gut. While SQDG inhibited the leucine amino peptidase and trypsin like serine protease activities, which decreased the content of total free amino acids. Imbalance in the activities of aspartate amino transferase and alanine amino transferase disrupted the aminoacid metabolism and the compound inhibited the activity of tyrosinase (an enzyme involved in cuticle development). SQDG toxicity caused accumulation of 20-hydroxyecdysone, the active moulting hormone in the hemolymph. Simultaneous action of two allomones present in Pisanglilin effectively resisted the attack of endophytic larvae in the pseudostem and thereby conferred resistance against infestation by O.longicollis. Preliminary study by intrapseudostem injection of Pisanglilin extract in susceptible M.paradisiaca cultivar Kappa, gave complete protection to it from attack by this pest, under field condition.

Phospholipid profiles and annexin V content in villous chorion from women with spontaneous miscarriage associated with cytomegalovirus infection

Aim. To carry out a comparative analysis of the phospholipid profiles and annexin V content in the villous chorion obtained from women with spontaneous abortion associated with exacerbation of cytomegalovirus infection (CMVI) and the physiological course of pregnancy.Materials and methods. 66 patients were randomly selected at a gestational age of 6-8 weeks. A control group was identified in the amount of 32 healthy pregnant women with medical abortion. The main study group consisted of 34 women with spontaneous abortion associated with exacerbation of cytomegalovirus CMVI. The material for the study was the villous chorion, peripheral blood and urine. Type-specific antibodies to CMV immunoglobulins M and G class and the avidity index were determined in blood serum by the method of enzyme-linked immunosorbent assay. Lipids in the villous chorion homogenate were extracted with a chloroform-methanol mixture, phospholipids were fractionated by thin layer chromatography. The quantitative determination of annexin V was carried out by the method of enzyme immunoassay.Results. The phospholipid profile of the villous chorion in the main group was characterized by a decrease in the fraction of phosphatidylcholine to 22,39±0,04% (control group – 28,50±0,052%; p<0,001), phosphatidylethanolamine to 27,63±0,011% (control group – 30,11±0,073%; p<0,001), phosphatidylinositol up to 14,24±0,021% (control group – 16,17±0,018%; p<0,001), an increase in lysophosphatidylcholine up to 2,25±0,032% (control group – 1,07±0,022%; p<0,001), phosphatidylserine up to 14,57±0,075% (control group – 10,02±0,012%; p<0,001), sphingomyelin up to 18,92±0,012% (control group – 14,13±0,012%; p<0,001). At the same time, an increase in the content of annexin V was revealed up to 43,12±0,50 U/mL (control group – 20,21±0,50 U/mL; p<0,001).Conclusion. The results of the study showed that in the villous chorion from women with spontaneous abortion associated with exacerbation of CMVI, there was a change in the profile and ratio of phospholipids, as well as in the concentration of annexin V. The established increase in the content of phosphatidylserine and the associated annexin V in the villous chorion indicates a violation of the hemostatic system and microcirculation due to increased thrombus formation, which supports local inflammation and apoptosis of the trophoblast, which leads to embryonic demise and spontaneous abortion.

Biological Activities of Different Strains of the Genus Ganoderma spp. (Agaricomycetes) from Mexico.

Antiproliferative, antioxidant, and antibacterial activities were determined for 14 extracts obtained with a mixture of chloroform-methanol (1:1) from the mycelial cultures of 14 wild strains of the genus Ganoderma collected in the central-south part of Veracruz Province, Mexico. Identification of the strains collected was confirmed based on rDNA internal transcribed spacer phylogenetic analysis. The strains G. tuberculosum (GVL-04 and GVL-21), G. tornatum (GVL-05), and G. weberianum (GVL-17 and GVL-26) manifested activity in at least one of the six cancer cell lines tested (HBL-100 and T-47D [breast], HeLa [cervix], A-549 and SW1573 [lung], and WiDr [colon]), with a minimum concentration necessary to cause 50% growth inhibition of cancer cells (GI50) < 50 μg/mL-1. The strains G. tuberculosum (GVL-21) and G. martinicense (GVL-35) had the best antioxidant activity, with values of 62.5 ± 3.9 and 40 ± 2.0 μM Trolox equivalents/mg according to the 1,1-diphenyl-2-picrihydrazyl assay. In addition, nine extracts demonstrated antibacterial activity against Clavibacter michiganensis in a concentration range of 31.5 to 1000 μg/mL. Although these results were expected due to the bioactive potential of Ganoderma species, the antibacterial activity against C. michiganensis causing tomato canker is highlighted.

Rapid determination of trace ciprofloxacin residue in milk samples using molecularly imprinted membrane extraction-high performance liquid chromatography-tandem mass spectrometry

An enrofloxacin (ENR) molecularly imprinted membrane (MIM) was prepared with a polyvinylidenedifluoride (PVDF) membrane as the carrier, ENR as the dummy template molecule, α-methacrylic acid (MAA) as the functional monomer, ethylene glycol dimethacrylate (EGDMA) as the cross-linker, and a chloroform-methanol mixture solvent as the porogen. The MIM showed excellent selectivity, high adsorption capacity, and high adsorption rate for ciprofloxacin. Additionally, a method combining molecularly imprinted membrane extraction (MIME) and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed and validated for the selective analysis of trace ciprofloxacin residue in milk samples. The sample pretreatment involved only a single step of protein precipitation. Ciprofloxacin showed good linearity in mass concentration range of 0.1-200 μg/L with a high correlation coefficient (r2>0.9996). The limit of detection (LOD, S/N=3) and limit of quantification (LOQ, S/N=10) were 0.02 μg/L and 0.1 μg/L, respectively. The relative standard deviations (RSDs) of interday and intraday precisions ranged from 3.3% to 7.9%. The ciprofloxacin recovery was in the range of 92.6%-119.1%. The results showed that the proposed method is simple and fast, with high accuracy and sensitivity, thus being suitable for the rapid detection of trace ciprofloxacin residue in milk samples.

Comprehensive Characterization and Relative Quantification of α-Amylase/Trypsin Inhibitors from Wheat Cultivars by Targeted HPLC-MS/MS.

The α-amylase/trypsin inhibitors (ATIs) are discussed as being responsible for non-celiac wheat sensitivity (NCWS), besides being known as allergenic components for baker’s asthma. Different approaches for characterization and quantification including proteomics-based methods for wheat ATIs have been documented. In these studies generally the major ATIs have been addressed. The challenge of current study was then to develop a more comprehensive workflow encompassing all reviewed wheat-ATI entries in UniProt database. To substantially test proof of concept, 46 German and Turkish wheat samples were used. Two extractions systems based on chloroform/methanol mixture (CM) and under buffered denaturing conditions were evaluated. Three aspects were optimized, tryptic digestion, chromatographic separation, and targeted tandem mass spectrometric analysis (HPLC-MS/MS). Preliminary characterization with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) documented the purity of the extracted ATIs with CM mixture and the amylase (60–80%)/trypsin (10–20%) inhibition demonstrated the bifunctional activity of ATIs. Thirteen (individual/common) biomarkers were established. Major ATIs (7–34%) were differently represented in samples. Finally, to our knowledge, the proposed HPLC-MS/MS method allowed for the first time so far the analysis of all 14 reviewed wheat ATI entries reported.

Synthesis and Spectral Properties of meso-Nitro-Substituted Octaethylporphyrins and Their Co(II) Complexes

The reaction of 2,3,7,8,12,13,17,18-octaethylporphyrin with sodium nitrite in trifluoroacetic acid produced 5-nitro-2,3,7,8,12,13,17,18-octaethylporphyrin, 5,15-dinitro-2,3,7,8,12,13,17,18-octaethylporphyrin, and 5,10,15-trinitro-2,3,7,8,12,13,17,18-octaethylporphyrin. Reactions of octaethylporphyrin, mono-, di-, tri-, and tetranitro-substituted porphyrins coordination to cobalt(II) acetate in a chloroform–methanol mixture were studied. The corresponding Co(II) porphyrinates were obtained and identified. It was shown that, upon dissolution of Co(II) octaethylporphyrinates in dimethylformamide in the presence of NaOH, the Co(II)→Co(III) oxidation occurs in the coordination site of the macrocycle.

Optimization of the manufacturing process of a complex amphotericin B liposomal formulation using quality by design approach.

In this work, the manufacturing process of a complex liposomal amphotericin B (AmB) product was optimized using quality by design (QbD) approach. A comprehensive QbD-based process understanding and design space (DS) to the critical process parameters (CPPs) is essential to the drug development and consistent quality control. The process was based on the acid-aided formation of drug-lipid complexes in a methanol-chloroform mixture (step I) followed by spray drying (step II), hydration and liposome formation by microfluidization (step III), and lyophilization (step IV). Firstly, the risk assessment was conducted to identify the critical process parameters among the four key steps. Nine CPPs and five CQAs (API Monomer identity (absorbance main peak at 321nm), API Aggregation identity (absorbance peak ratio, OD 415nm/321nm), particle size, in-vitro toxicity, and the cake quality) were determined based on their severity and occurrences with their contribution to the quality target product profile (QTPP). Based on the risk assessment results, the final screening design of experiments (DoE) was developed using fractional factorial design. Secondly, the empirical equation was developed for each CQA based on experimental data. The impact of CPPs on the CQAs was analyzed using the coefficient plot and contour plot. In addition to the effect of individual formulation parameters and process parameters, the effects of the four key separate steps were also evaluated and compared. In general, the curing temperature during microfluidization has been identified as the most significant CPP. Finally, design space exploration was carried out to demonstrate how the critical process parameters can be varied to consistently produce a drug product with desired characteristics. The design space size increased at the higher value of the curing temperature, the API to phospholipid ratio (API:PL), and the lower value of the DSPG to phospholipid ratio (PG:PL) and aspirator rate.

Separation of methanol-chloroform mixture using pressure-swing distillation: Modeling and optimization

The separation of methanol-chloroform mixture, a minimum-boiling azeotrope, is performed using pressure- swing distillation process via process simulation. In this study, the steady-state optimization was carried out using PRO/II with PROVISION v.10. The two different column configurations (low-to-high pressure and high-to-low pressure) were compared wherein the positions of the low-pressure column and high-pressure column were operated interchangeably to attain an optimized design. Additionally, different heat-integration configurations (partial heat- and full heat-integration) were applied to lessen the overall utility consumption. It was determined that the low-to-high pressure column configuration provided a more optimized result for all heat-integrated systems as compared to high-tolow pressure column configuration. Application of heat-integration further decreases the cooling water and steam consumption by 38.86% and 35.74%, respectively, for partial heat-integrated system, and by 44.58% and 41.01%, respectively, for full heat-integrated system.

Characterization of two Moroccan watermelon seeds oil varieties by three different extraction methods

The purpose of this study is to assess and evaluate the physicochemical properties of the seed oils of two Moroccan varieties of watermelon “Citrullus lanatus” extracted by three different techniques, a mechanical process using cold press, and two chemical processes using a Soxhlet apparatus and a sonotrode ultrasound assisted extraction (UAE) using n-hexane. The total phenolic compounds (TPC) and antioxidant properties against the DPPH radicals (2,2-diphenyl-1-picrylhydrazyl) were also studied. The seed oils of both varieties of watermelon exhibited high concentrations of unsaturated fatty acids with the predominance of linoleic and oleic fatty acids. The primary sterol was β-sitosterol, and high levels of total tocopherols were observed. Ɣ-tocopherol was the predominant tocopherol in all tested oils. TheCitrullus lanatus var. lanatusvariety seeds oil exhibited the highest TPC value (89.5 ± 0.06 mg EGA/100 g; EGA: gallic acid equivalent), with 82.4 ± 0.03% DPPH free radical inhibition efficiency. Nevertheless, all tested seeds oils showed a significant amount of total phenolic compounds and a good inhibition against DPPH radicals ranging from 51.1 ± 0.1% to 84.8 ± 0.04%. In addition, the influence of the ultrasonic extraction parameters was studied using two different solvents (n-hexane and the methanol-chloroform mixture), with different particle sizes (500–300 µm), duration (10 and 20 minutes), cycle (1–0.5), amplitude (80–100%) and solvent/seed ratios (1:5 and 1:10), and the seed roasting parameter was also studied. The oil yield was mainly affected by the extraction solvent, then the solvent/seed ratio and the duration, respectively.

Enzymatic Quantification of Liver Lipids After Folch Extraction.

The determination of the lipid content in liver offers the potential to investigate metabolic alterations in different research contexts. Here, we describe a method to determine cholesterol, triglycerides, and phospholipids in liver samples based on total lipid isolation by a 2:1 chloroform-methanol mixture (Folch extraction) and specific enzymatic colorimetric microassays in plate.

Adipose tissue-derived stem cells upon decellularized ovine small intestine submucosa for tissue regeneration: An optimization and comparison method.

The extracellular matrix of different mammalian tissues is commonly used as scaffolds in the field of tissue engineering. One of these tissues, which has frequently been studied due to its structural and biological features, is the small intestine submucosal membrane. These research are mainly done on the porcine small intestine. However, a report has recently been published about a scaffold produced from the submucosal layer of the ovine small intestine. In the present study, ovine small intestine submucosal (OSIS) was decellularized in a modified manner and its histological, morphological, and biomechanical properties were studied. Decellularization was performed in two phases: physical and chemical. In this method, a chloroform-methanol mixture, enzymatic digestion, and a constant dose of sodium dodecyl sulfate (SDS) was used in the least agitation time and its histological property and biocompatibility were evaluated in the presence of adipose tissue-derived stem cells (ADSCs); furthermore, ADSCs were isolated with a simple method (modified physical washing non-enzymatic isolation). The results were showed that the use of OSIS could be effective and operative. Mechanical properties, histological structure and shape, and glycosaminoglycan content were preserved. In the SDS-treated group, more than 90% of the native cells of tissue were deleted, and also in this group, no toxicity was observed and cell proliferation was supported, compared to the untreated group. Therefore, our results indicate that ADSCs seeded on OSIS scaffold could be used as a new approach in regenerative medicine as hybrid or hydrogel application.

Жирнокислотний склад тканин, відтворна здатність і медова продуктивність бджіл за згодовування соняшникової олії

The purpose. To position the link between fat-acid content of tissues and productive attributes of honeybees at different amount of sunflower seed oil in feed additive. To determine replicated ability of queen bees and honey productivity of workers. Methods. Probe of breeding capacity of queens was spent according to F.A. Lavrekhin and S.V. Pankova procedure. Amount of the gained commodity honey they determined by method of weighting honey taken from honeycomb jacks before and after pump down. In feed additive and tissues of honeybees they determined presence and content of fatty acid of common lipids using method of gas-liquid chromatography (I.F. Rivis with co-authors). The content of fatty acids of common lipids in feed additive and probed tissues of bees they determined by method of extraction of lipids by chloroform-methanol mixture. Results. Feeding of feed additive, enriched with sunflower seed oil in amount of 10 and 20 g essentially increased concentration of saturated, monononsaturated and polynonsaturated fatty acids of common lipids in tissues of abdomen, chest and head of honeybees of the Ist and especially the IInd probed groups. It testified to substantial growth of security of tissues of honeybees with power and structural materials. Thus in probed tissues of bees the ratio of content of polynonsaturated fatty acids of family u-3 (linolenic row) to polynonsaturated fatty acids of family --6 (linolic row) was diminished. Conclusions. Changes in fat-acid content of tissues of abdomen, chest and head of honeybees of the Ist and especially the IInd probed groups were accompanied by changes of breeding capacity of queens and honey productivity of workers. In particular at queens of these groups grew ovi-production, and at workers — honey productivity.