Synthesis of novel fullereneα-amino acid conjugates

Aspartic acid and glutamic acid with protected α-amino and α-carboxyl groups had been used to react with the activated hydroxyl group of N-substituted 3,4-fullero pyrrolidine. The products were deprotected, affording two monofuUerene α-amino acids, monofullerene aspartic acid （mFas） and monofullerene glutamic acid （mFgu）. Then a bifullerene glutamic acid conjugate （bFguC） was synthesized by reaction of mFgu containing protected amino group with N-substituted 3,4-fullero pyrrolidine.

Combretastatin A4/poly(L-glutamic acid)-graft-PEG conjugates self-assembled to nanoparticles

Combretastatin A4(CA4) possesses varying ability to cause vascular disruption in tumors,while the short half-life, low water solubility and deactivation of many CA4 analogs during storage limited its antitumor efficacy and drug stability. A novel macromolecular conjugate of CA4(CA4-PL) was synthesized by covalent bonding of CA4 onto poly(L-glutamic acid)-graft-polyethylene glycol(PLG-g-PEG) via Yamaguchi reaction. The obtained CA4-PL was characterized by ~1H NMR, GPC, and UV methods, and the properties of the nanoparticles composed of CA4-PL, including critical aggregation concentration, size and size distribution, and morphology, were investigated. CA4-PL can self-assemble to form micelle-like nanoparticles of 80～120 nm in diameter, which may have potential to improve the blood circulation period as well as the targetability of CA4, and find applications to treat various tumors when combined with traditional chemotherapy or radio therapy.

Preparation of Conjugated Linoleic Acid of High Purity and Its Apparent Kinetic Characteristics During Formation

Conjugated linoleic acid (CLA) is a fatty acid with physiological activities and potential application prospect. This paper focuses on the method of synthesis of conjugated linoleic acid of high purity and the process line and conditions for its purification that can be used in large scale production. CLA of more than 95% purity was prepared by means of urea adduct purification and conjugation using safflower oil as material. The total recovery of the product adds up to more than 48%. The reactive kinetics about linoleic acid from sunflower oil converted into CLA was investigated, and its apparent kinetic model was also established, which can be used as a base for industrial designs.

Preparation of Conjugated Linoleic Acid and Identification of Its Isomers

Conjugated linoleic acid (CLA) is a kind of fatty acid with physiological activities and potential application prospect. A synthesis method of conjugated linoleic acid and a purification technology were studied. CLA was prepared and purified by urea-complexation and conjugation using safflower oil as raw material. The purity of CLA and total recovery of the product was more than 95% and 48%, respectively. The main isomers produced in alkali-catalyzed conjugation were identified by gas chromatography (GC) linked to mass spectrometry (MS) and Fourier transform infrared spectroscopy (FTIR). The total amount of the two main isomers (9cis, 11trons-and 10trans, 12cis-CLA) determined by GC was more than 90% of the product.

Effect of cis-9,trans-11-conjugated linoleic acid on cell cycle of gastric adenocarcinoma cell line(SGC-7901)

AIM: To determine the effect of cis -9, trans -11-conjugated linoleic acid (c9, t11-CLA) on the cell cycle of gastric cancer cells (SGC-7901) and its possible mechanism in inhibition cancer growth. METHODS: Using cell culture and immunocytochemical techniques, we examined the cell growth, DNA synthesis, expression of PCNA, cyclin A, B(1), D(1), p16(ink4a) and p21(cip/waf1) of SGC-7901 cells which were treated with various c9, t11-CLA concentrations (25, 50, 100 and 200 micromol.L(-1))of c 9, t 11-CLA for 24 and 48h, with a negative control (0.1% ethane). RESULTS: The cell growth and DNA synthesis of SGC-7901 cells were inhibited by c9, t11-CLA.SGC-7901 cells. Eight day after treatment with various concentrations of c9, t11-CLA mentioned above, the inhibition rates were 5.92%, 20.15%, 75.61% and 82.44%, respectively and inhibitory effect of c9, t11-CLA on DNA synthesis (except for 25 micromol.L, 24h) showed significantly less (3)H-TdR incorporation than that in the negative controls (P【0.05 and P【0.01). Immunocytochemical staining demonstrated that SGC-7901 cells preincubated in media supplemented with different c9, t11-CLA concentrations at various times significantly decreased the expressions of PCNA (the expression rates were 7.2-3.0%, 24h and 9.1-0.9% at 48h, respectively), Cyclin A (11.0-2.3%, 24h and 8.5-0.5%,48h), B(1) (4.8-1.8% at 24h and 5.5-0.6% at 48h)and D(1) (3.6-1.4% at 24h and 3.7%-0 at 48h) as compared with those in the negative controls(the expressions of PCNA, Cyclin A, B(1) and D(1) were 6.5% at 24h and 9.0% at 48h, 4.2% at 24h and 5.1% at 48h, 9.5% at 24h and 6.0% at 48h,respectively)(P【0.01), whereas the expressions of P16(ink4a) and P21(cip/waf1), cyclin-dependent kinases inhibitors(CDKI), were increased. CONCLUSION: The cell growth and proliferation of SGC-7901 cell is inhibited by c9, t11-CLA via blocking the cell cycle, with reduced expressions of cyclin A,B(1) and D(1) and enhanced expressions of CDKI(P16(ink4a) and p21(cip/waf1)).

Chiral Bronsted acid-catalyzed conjugate addition of indoles to azadienes:Enantioselective synthesis of hetero-triarylmethanes

An efficient chiral Br?nsted acid-catalyzed conjugate addition of indoles to azadienes has been successfully developed,which enables a facile approach to optically active hetero-triarylmethanes with excellent enantioselectivities and broad substrate scope.This chiral Br?nsted acid catalytic system provides a new opportunity for the development of asymmetric reactions of azadienes.

Influence of dietary conjugated linoleic acid on growth, fatty acid composition and hepatic lipogenesis in large yellow croaker (Pseudosciaena crocea R.)

We examined the effects of conjugated linoleic acid (CLA) on growth, fatty acid composition and enzyme activity of fatty acid oxidation in the liver of large yellow croaker. We divided 1600 fish (average initial weight 150 g) into 4 groups and reared them in 8 cages. Four dietary treatments were formulated to contain 0%, 1%, 2% and 4% (w/w) CLA, respectively. The fish were fed for 10 weeks ad libitum twice daily. We found that the dietary CLA had no effect on growth, biometric parameters and whole body proximate (P>0.05), but showed some significant effects on the fatty acid composition in both muscle and the liver. The activities of lipogenic enzymes were slightly depressed in fish fed with increasing levels of CLA when compared with control (P>0.05). Dietary CLA supplementation had no effects on liver lipid content, but significantly increased the contents of saturated fatty acids (SFA) and polyunsaturated fatty acids (PUFA) (P<0.05) and decreased monounsaturated fatty acid (MUFA) content in both muscle and the liver. Dietary CLA inclusion resulted in significant increases of the biologically active cis-9, trans-11 and trans-10, cis-12 isomers in both tissues (P<0.05). The total accumulation of CLA was higher in the liver (3.83%, w/w) than in muscle (3.77%, w/w) when fed with 4% (w/w) CLA. This study demonstrates that large yellow croakers are capable of absorbing and depositing CLA and long-chain n-3 PUFA in the liver and muscle, showing that this species fed with CLA could be an important human food source for these healthful fatty acids.

The Effect of Parity on Conjugated Linoleic Acid(CLA) Content in Milk Fat and CLA-Desaturase Index from Holstein Dairy Cows

The performance of lactating dairy cows and composition of the fatty acids in milk fat were measured to investigate the effect of parity on conjugated linoleic acid(CLA)in milk fat and△~9-desaturase indices from Holstein dairy cows.Milk samples of two hundred and thirty-five lactating Holstein dairy cows in north China were tested.In order to avoid possible confounding effects of diet and season,Holstein cows were fed a single diet and milk was sampled on the same day.The average content of cis-9,trans-11 CLA was 5.14 mg/g fatty acids in milk fat,varying between 1.5 and 10.5 mg/g fatty acids.The average content of cis-9,trans-11 CLA of primiparous cattle was 5.19 mg/g fatty acids in milk fat,while it was 5.11 mg/g fatty acids in multiparous animals,with no significant difference between them(P】0.05).There was a significant(P【0.05)difference between the△~9-desaturase index of cis-9 C14:l and cis-9 C16:l between primiparous and multiparous cattle but not(P】0.05)in the△~9-desaturase index of cis-9 C18:l and cis-9,trans-11 CLA.

Conjugated Linoleic Acid and Dietary Fats Differentially Affect Hepatic ACAT Activity and LDL-Cholesterol in Postweanling Pigs Fed Low-Fat Diets

ABSTRACT：Two separate studies tested the hypoth- esis that plasma low-density lipoprotein cholesterol LDL-C ） can be decreased by conjugated linoleic acid （CLA） by depressing hepatic acyl-coenzyme A： cholesterol acyltransferase （ACAT） activity. In the first experiment, 3 groups of 6 early-weaned piglets were fed low-fat diets containing either 1.5% CLA, 1.5% corn oil or 1.5% beef tallow;fat provided 8% of the energy intake. In the second experiment, 4 groups of 6 early-weaned piglets were fed high-fat di- ets containing either 15% beef tallow, 12% beef tal- low plus 3% CLA, 15% corn oil, or 12% corn oil plus 3% CLA; fat provided 29% of energy intake. Cholesterol was balanced across diets in both experi-ments. In pigs fed the low-fat diets, all dietary fats in- creased LDL-C and triacylglycerols and decreased high-density lipoprotein cholesterol （ HDL-C ） and very low-density lipoprotein cholesterol （VLDL-C）. LDL-C was the same in pigs fed low-fat tallow or low-fat CLA diets. However, ACAT activity was near- ly 80% higher in pigs fed the low-fat tallow diet than in pigs fed the low-fat CLA diets. All high-fat diets increased LDL-C, HDL-C and triacylglycerols equally with no effect on VLDL-C. There were no unique fat- ty acid effects of the high-fat diets on ACAT activity. We conclude that supplemental fats had differential effects on hepatic ACAT activity and LDL-C, but on- ly in pigs fed low-fat diets.

A Novel Thin Film Resistive Humidity Sensor Based on SolubleConjugated Polymer:(propionic acid)-co-(propargyl alcohol)

A novel soluble conjugated copolymer (propionic acid)-co-(propargyl alcohol) (PA-co-OHP) has been synthesized for the first time using a new palladium acetylide catalyst Pd(PPh3)(2)(C=CC(CH3)(2)OH)(2)(PPB). Thin film resistive humidity sensor based oil the copolymer doped with HClO4 was prepared. The impedance of the sensor changed from 10(3)similar to 10(7) Omega in 95%similar to 30%RH, and the response of that is very quick (<6 sec.). Preliminary results show the copolymer is a promising humidity sensitive material.

Effects of Conjugated Linoleic Acids on Proliferation and Differentiation of Rat Preadipocytes

Conjugated linoleic acids （CLAs） are a generic term for linoleic acid isomers and have a variety of biological functions. After rat preadi- pocytes were incubated in CLAs-supplemented media, their proliferation and differentiation were observed by the cell count and the oil red O stai- ning. The results showed that the CLAs at different concentrations inhibited proliferation of the rat preadipocytes in a time- and dose-dependent manner. And the CLAs greatly decreased intracellular lipid content in mature adipocytes. Moreover, lipogenesis was inhibited by the CLAs in a dose-dependent manner. Therefore, the CLAs inhibit the lipogenesis by reducing the number of preadipocytes and decreasing the intracellular lipid

Optimization of conjugated linoleic acid triglycerides via enzymatic esterification in no-solvent system

We compared four esterifiable enzymes. The lipase Novozym 435 possessed the highest activity for the conjugated linoleic acid esterification during the synthesis of triglycerides. The triglycerides were synthesized by esterification of glycerol and conjugated linoleic acid （CLA） in a no-solvent system using lipase catalysis. We investigated the effects of temperature, enzyme concentration, water content, and time on esterification. Enzyme and water concentrations of up to 1% of the total reaction volume and a system temperature of 60℃ proved optimal for esterification. Similarly, when the esterification was carried out for 24 h, the reaction ratio improved to 94.11%. The esterification rate of the rotating screen basket remained high （87.28%） when the enzyme was re-used for the 5th time. We evaluated the substrate selectivity of lipase （NOVO 435） and determined that this lipase prefers the 10,12-octadacadienoic acid to the 9,11-octadecadienoic acid.

Hydrolysis of soy isoflavone conjugates using enzyme may underestimate isoflavone concentrations in tissue

Objective:To investigate the differences of using enzymatic hydrolysis and acid hydrolysis for identification and quantification of isoflavone aglycones from biomatrices.Methods:β-glucuronidase/sulfatase isolated from Helix pomatia for routine enzymatic hydrolysis or 6N HCl was used to release glucuronide and sulfate conjugates in the serum,urine and tissue samples.Profiles of soy isoflavones after enzymatic hydrolysis or acid hydrolysis in several tissues of rat fed with diets containing soy protein isolate were also compared using LC/MS and HPLC-ECD.Results:Acid hydrolysis released more aglycone than enzymatic digestion(P<0.05)in liver tissue.The total genistein,daidzein and other metabolites were 20%to 60%lower in samples from enzymatic hydrolysis than in acid hydrolysis.Conclusion:These results indicated that unknown factors in tissues reduced the enzymatic hydrolytic efficiency for releasing isoflavone aglycones even in optimized condition.This would underestimate isoflavone tissue concentrations up to 60%.

Increasing Conjugated Linoleic Acid Content in Milk and Cheese after Supplementing a Blend of Crude Soybean Oil Sediment Combined with Fish Oil to Grazing Dairy Cows

The aim of the work was to improve the healthy value of milk and cheese fatty acids (FA) by feeding a mix of crude soybean oil sediment (CSOS) combined with fish oil (FO) to grazing dairy cows. The CSOS is a by-product commonly discarded after oil extraction containing 3.3% moisture, 6% total ash and 70.7% oil, locally available, comparatively economic and easy to mix with other feed ingredients. The experiment lasted 55 days from September 30th to November 23th 2018 and was carried out at the dairy farm “Gacef” provider of milk to the dairy industrial plant “Capilla Del Señor” (CDS) located at the Villa María City, Córdoba Province, Argentine. A herd of 80 multiparous Holstein cows producing 24 kg-1 milk·cow-1·day-1 was used. The cows grazed an alfalfa and an oat pasture that represented about 47% of total dry matter (DM) intake supplemented at 8.5 kg DM·cow-1·day-1 with a total mixed ration (TMR) composed (DM basis) by cracked corn grain (35.18%), whole plant corn silage (31.98%), pelletized soyben meal (17.99%), the CSOS supplement (13.85%) and FO (0.99%). The TMR was supplied by halves after each milking time in groupal feeders yielding 1.4 kg·cow-1·day-1 of the CSOS and 0.1 kg·cow-1·day-1 of FO. Before the start of lipid supplementation, milk samples (5) were obtained from the farm-tank representing the standar or reference milk (Ref-Milk). After 21 days of supplementary lipid supply, additional milk samples (5) were obtained representing the modified milk (Mod-Milk). Milk samples were analyzed for chemical composition and milk FA profile. At each time, sufficient quantities of both (Ref- and Mod-Milk) were collected for manufacturing six types of cheeses. The results were analyzed through the Student-T test for independent observations. Oil supplementation did not modify (P > 0.05) the chemical composition of milk. Concentration of butyric acid (C4:0) in milk was not affected (P < 0.858). Concentration of total saturated FA (SFAs) in Ref-Milk averaged 58.83 g 100 g-1 FA and was decreased to 49.67 g 100 g-1 FA in Mod-Milk (P < 0.0001). Monounsaturated FA (MUFAs) increased (P < 0.001) from 32.03 g 100 g-1 FA in Ref-Milk to 38.13 g 100 g-1 FA in Mod-Milk (+19.07%) whereas polyunsaturated FA (PUFAs) increased (+36.1%) from 4.71 to 6.41 (P < 0.004). The Mod-Milk showed a significant (P < 0.002) reduction (-15.3% or 5.9 g 100 g-1 FA) for the total concentration of the potentially atherogenic fraction of milk FA (C12:0 to C16:0). The atherogenic index (AI) also decreased (P < 0.012) from 1.98 in Ref-Milk to 1.42 in Mod-Milk (-28.4%). Concentration of vaccenic acid (VA, trans-11 C18:1) in Mod-Milk averaged 7.77 g 100 g-1 FA which represented a 162 % increase (P < 0.0001) over that observed in Ref-Milk (2.95 g 100 g-1). Concentration of conjugated linoleic acid (CLA, cis-9, trans-11 C18:2) in Ref-Milk averaged 1.47 g 100 g-1 FA and showed an important increase (P < 0.002) in the Mod-Milk (3.86 g 100 g-1 FA, +163%). The omega 6/3 ratio resulted lower (P < 0.012) in the Ref-Milk (2.28) compared to the Mod-Milk (2.83). Milk and cheese FA composition were highly correlated (R2 = 0.99, P < 0.0001). The Mod-Cheeses showed similar results in AI, total concentration of SFAs, MUFAs and PUFAs compared to the milk of origin. Differences in FA composition between the cheeses made with the Ref- and Mod-Milk were equivalent to those described for milks. It is concluded that supplementation with a blend of CSOS supplement and FO was an effective way to improve the healthy value of dairy products by reducing contents of SFAs, atherogenic FAs and the atherogenicity index with a concomitant increase in VA and CLA. Modifications induced in the Mod-Milk were recovered in the Mod-Cheeses. The results obtained may help to reduce saturated fat intake and fight or prevent incidence of non-communicable, cardiovascular and chronic diseases.

Tauroursodeoxycholic acid:more than just a neuroprotective bile conjugate

Neurological diseases and the neuroinflammatory response：Neurological diseases are usually accompanied by dramatic changes in the tissue homeostasis,inducing a neuroinflammatory environment that leads to the progressive activation of central nervous system（CNS）resident cells.

Effect of Supplementation of Conjugated Linoleic Acid in Diets on Growth Performance and Total Lactic Bacteria iin Small Intestine of Broiler

The effect of supplementation of conjugated linoleic acid （CLA） in diets on growth performance and total lactic bacteria （LAB） in small intestine of broiler. One hundred sixty eight broilers （Arbor Acres; 1 wk old） were randomly assigned to 8 groups. Each group （male 21 and female 21 broiler chickens） was further randomly divided into 3 replicates of 7 chickens were assigned to 4 dietary treatments （0%, 0.5%, 1.0%, and 1.5% CLA）. Feed consumption and BW were recorded at every 7-day period. On day 56, and each week were collected at ileal content in small intestine for the total count of lactic acid bacteria （LAB）. There were highly significant differences in body weight gain, average dairy feed intake, average dairy gain and feed conversion ratio among the treatments （P 〈 0.01）. Moreover, our results revealed that the highest detection trend from the ileal occurred during the period in the supplementation of CLA.

EFFECT OF CIS-9, TRANS-11-CONJUGATED LINOLEIC ACID ON CELL CYCLE OF MAMMARY ADENOCARCINOMA CELLS(MCF-7)

Objective: To determine the effect of cis-9, trans-1 1-conjugated linoleic acid on the cell cycle of mammary cancer cells (MCF-7) and the possible mechanism of the inhibitory effect of c9,t11-CLA. Methods: Using cell culture and immunocytochemical techniques, we examined the cell growth, DNA synthesis, expression of PCNA, cyclin A, B1, D1, p16ink4a and p21cip/waf1 of MCF-7 cells at various c9,t11-CLA concentrations (25μM, 50μM, 100μM and 200μM), at 24h and 48h. 96% ethand was used as negative control. Results: The cell growth and DNA synthesis of MCF-7 cells were inhibited by c9,t11-CLA. After treatment with various doses of c9,t11-CLA mentioned above for 8 days, the inhibition frequency was 27.18%, 35.43%, 91.05%, and 92.86%, respectively. Inhibitory effect of c9,t11-CLA on DNA synthesis (except for 25μM, 24h) was demonstrated by significantly less incorporation of 3H-TdR than the negative control (P<0.05 and P<0.01). To further investigate the influence of the cell cycle progression, we found that c9,t11-CLA may arrest the cell cycle of MCF-7 cells. Immunocytochemical staining demonstrated that incubation with different concentration of c9,t11-CLA at various times significantly decreased the expression of PCNA, Cyclin A, B1, D1 in MCF-7 cells compared to the negative control (P<0.01), whereas the expression of p16ink4a and p21cip/waf1, cyclin-dependent kinases inhibitors (CDKI), were increased. Conclusions: The cell growth and proliferation of MCF-7 cells is inhibited by c9,t11-CLA via blocking cell cycle, accompanying reduced expression of cyclin A, B1, D1 and enhanced expression of CDKI (p16ink4a and p21cip/waf1).

Study on Microwave Preparation of Conjugated Linoleic Acid from Corn Germ Oil

[Objective] The optimal craft of preparing conjugated linoleic acid from corn germ oil by dint of microwave method was studied. [Meth- od] Microwave power, microwave mode of action, the amount of catalyst and solvent were selected for single-factor-experiment. Then, based on the single-factor-experiment, the catalyst and solvent amount, and microwave function time were chosen for orthogonal experiment so as to deter- mine the optimal craft of preparing CLA from corn germ oil. UV absorption spectra and infrared absorption spectrum were used to analyse the struc- tural features of natural corn germ oil and CLA. [Resalt] The optimal preparation condition was as follow： the catalyst was 0.6 times heavier than oil, solvent was 3 times heavier than oil weight, microwave worked for ten minutes. Under the optimal condition, CLA was 56.3% and conversion rate was 99.47 %. The results indicated that non-conjugated double bonds of linoleic acid can be isomerized and changed into conjugated double bonds under the conditions of microwave irradiation,[ Conduslon] The study provided a new way for the preparation of CLA.

Folate-conjugated Fe_3O_4 nanoparticles for in vivo tumor labeling

Highly biocompatible superparamagnetic Fe3O4 nanoparticles were synthesized by amide of folic acid （FA） ligands and the NH2-group onto the surface of Fe3O4 nanoparticles. The as-synthesized folate-conjugated Fe3O4 nanoparticles were characterized by X-ray diffraction diffractometer, transmission electron microscope, FT-IR spectrometer, vibrating sample magnetometer, and dynamic light scattering instrument. The in vivo labeling effect of folate-conjugated Fe3O4 nanoparticles on the hepatoma cells was investigated in tumor-bearing rat. The results demonstrate that the as-prepared nanoparticles have cubic structure of Fe3O4 with a particle size of about 8 nm and hydrated diameter of 25.7 nm at a saturation magnetization of 51 A·m2/kg. These nanoparticles possess good physiological stability, low cytotoxicity on human skin fibroblasts and negligible effect on Wistar rats at the concentration as high as 3 mg/kg body mass. The folate-conjugated Fe3O4 nanoparticles could be effectively mediated into the human hepatoma Bel 7402 cells through the binding of folate and folic acid receptor, enhancing the signal contrast of tumor tissue and surrounding normal tissue in MRI imaging. It is in favor of the tumor cells labeling, tracing, magnetic resonance imaging （MRI） target detection and magnetic hyperthermia.