Effects of chitosan nanoparticles on seed germination and seedling growth of wheat (Triticum aestivum L.).

To investigate the effect and mechanism of chitosan nanoparticles (CSNPs) on the germination and seedling growth of wheat (Triticum aestivum L.), we conducted systematic research on the impact of different concentrations (1-100 µg/mL) of CSNPs and chitosan (CS). The result of energy-dispersive spectroscopy (EDS) and confocal laser scanning microscopy (CLSM) showed that adsorption of CSNPs on the surface of wheat seeds was higher than that of CS. CSNPs had growth promoting effect at a lower concentration (5 µg/mL) compared with CS (50 µg/mL). In addition, the application of 5 µg/mL CSNPs induced the auxin-related gene expression, accelerated indole-3-acetic acid (IAA) biosynthesis and transport, and reduced IAA oxidase activity resulting in the increase of IAA concentration in wheat shoots and roots. The results suggest that CSNPs have positive effect on seed germination and seedling growth of wheat at a lower concentration than CS due to higher adsorption on the surface of wheat seeds.

Activation of Macrophages in vitro by Phospholipids from Brain of Katsuwonus pelamis (Skipjack Tuna).

The biological activities of phospholipids (PLs) have attracted people's attention, especially marine phospholipids with omega-3 polyunsaturated fatty acids DHA and EPA. In this study, we investigated the immunity activation of macrophages in vitro by phospholipids from skipjack brain. The phospholipids were extracted with hexane and ethanol ultrasonication instead of the traditional method of methanol and chloroform. The content of phospholipids from Skipjack brain was 19.59 g/kg by the method (the ratio of hexane and ethanol 2:1, 40 min, 35°C, 1:9 of the ratio of material to solvent, ultrasonic power 300W, ultrasonic extraction 2 times). The RAW264.7 macrophages were stimulated by the phospholipids from the Skipjack, by which the volume, viability and phagocytosis of macrophages were increased. The concentration of NO and the activity of SOD of the cells were also enhanced. The gene expressions of IL-1ß, IL-6, iNOS and TNF-α mRNA assayed by RT-PCR were up-regulated. Phospholipids from brain of Skipjack Tuna could activate macrophages immunity which displayed to induce pro-inflammatroy cytokines mRNA expression.

Effects of Calcium Alginate Submicroparticles on Seed Germination and Seedling Growth of Wheat (Triticum aestivum L.).

Calcium alginate (CaAlg) submicroparticles have a potential application in agricultural delivery systems. This study investigated the effects of CaAlg submicroparticles on seed germination and seedling growth of wheat. CaAlg submicroparticles with a Z-average diameter of around 250.4 nm and a measured zeta potential value of about -25.4 mV were prepared and characterized by dynamic light scattering (DLS), scanning electron microscopy (SEM) and energy dispersive X-ray spectrometer (EDS). After this, the effects of the concentration of CaAlg submicroparticles (10⁻500 µg/mL) on germination percentage, seedling length, the number of adventitious roots, chlorophyll content and soluble protein content were evaluated. The results demonstrated a significant increase in the level of germination percentage (9.0%), seedling index (50.3%), adventitious roots (27.5%), seedling length (17.0%), chlorophyll (8.7%) and soluble protein contents (4.5%) at a concentration of 100 µg/mL. However, an inhibitory effect was observed at a concentration of 500 µg/mL. The SEM examination showed that CaAlg submicroparticles could be successfully adsorbed onto the surface of the wheat seed. Further studies proved that CaAlg submicroparticles at a concentration of 100 µg/mL promoted the expression of indole-3-acetic acid (IAA)-related genes (YUCCA9, AUX1, ARF and UGT) in wheat, which resulted in an increase of 69% and 21% in IAA concentration in wheat roots and shoots, respectively.

[Synthesis and biological activities of beta-chain fragments of hemoglobin].

To investigate the angiotensin I-converting enzyme (ACE) inhibitory activity of beta-chain hemoglobin fragments, 17 fragments were synthesized by microwave-assisted solid-phase synthesis method. Wang resin or Trt(2-Cl) resin, Fmoc and HBTU-HOBt were used as solid carrier, N-terminal amino acid protecting groups and coupling reagents, respectively. The ACE inhibitory, alpha-glucosidase inhibitory, antibacterial and antitumor activities of the synthesized fragments were assayed. In vitro, Val-Val-Tyr-Pro-Trp-Thr showed high ACE inhibitory activity (IC50 = 7.42 micromol x L(-1)). The results indicate that there are two active sites in Val-Val-Tyr-Pro-Trp-Thr-Gln-Arg-Phe, one consists of Val-Val-, and the other -Gln-Arg-Phe. Peptides showed high ACE inhibitory activity when the N-terminal was hydrophobic amino acid such as Val and C-terminal tripeptide contained Phe, Trp or Arg. Some of the fragments showed low a-glucosidase inhibitory activity. No antibacterial activity or antitumor activity was detected in vitro. The results indicate that these peptides have a potential antihypertensive effect and possible application in the treatment of hypertension.

Purification and identification of a ACE inhibitory peptide from oyster proteins hydrolysate and the antihypertensive effect of hydrolysate in spontaneously hypertensive rats.

Oyster (Crassostrea talienwhanensis Crosse) proteins were produced from fresh oyster and subsequently digested with pepsin. The separations were performed with a Sephadex LH-20 gel filtration chromatography and a RP-HPLC. A purified peptide with sequence Val-Val-Tyr-Pro-Trp-Thr-Gln-Arg-Phe (VVYPWTQRF) was firstly isolated and characterized from oyster protein hydrolysate and its ACE inhibitory activity was determined with IC50 value of 66µmol/L in vitro. Stability study for ACE inhibitory activity showed that the isolated nonapeptide had the good heat and pH stability and strong enzyme-resistant properties against gastrointestinal proteases. Kinetic experiments demonstrated that inhibitory kinetic mechanism of this peptide was non-competitive and its Km and Ki values were calculated. The yield of this peptide from oyster proteins was 8.5%. Furthermore, the oyster protein hydrolysate (fraction II), prepared by pepsin treatment firstly exhibited antihypertensive activity when it was orally administered to spontaneously hypertensive rat (SHR) at a dose of 20mg/kg. These results demonstrated that the hydrolysate from oyster proteins prepared by pepsin treatment could serve as a source of peptides with antihypertensive activity.

Isolation and characterization of angiotensin I-converting enzyme inhibitory peptides derived from porcine hemoglobin.

Animal blood is potentially an untapped source of drugs and value-added food production. More than 400 million pigs are slaughtered each year but porcine blood is usually discarded in China. This study describes the isolation and characterization of angiotensin I-converting enzyme (ACE) inhibitory peptides derived from porcine hemoglobin. The most active hydrolysate was obtained from the peptic digestion of porcine hemoglobin. After the purification of ACE-inhibitory peptides with Sephadex LH-20 gel chromatography and reversed-phase high-performance liquid chromatography (RP-HPLC) on C(18) column, two active fractions were obtained. They were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS). They were LGFPTTKTYFPHF and VVYPWT, corresponding to the 34-46 fragment of the alpha chain and the 34-39 fragment of the beta chain of porcine hemoglobin, with IC(50) values of 4.92 and 6.02 microM, respectively. They were the first found from porcine hemoglobin; in particular, LGFPTTKTYFPHF was a novel ACE-inhibitory peptide. In addition, the purified ACE inhibitors both competitively inhibited ACE, and maintained inhibitory activity even after incubation with gastrointestinal proteases. This suggests that these peptides might have a potential antihypertensive effect.

A synthetic polymer, poly(2-methacryloyloxyethyl phosphorylcholine-co-n-stearyl methacrylate), stimulates insulin release from RINm5F insulinoma cells.

A water-soluble phospholipid-like polymer, poly(2-methacryloyloxyethyl phosphorylcholine-co-n-stearyl methacrylate) (PMC(18), average molecular weight = 4.3 x 10(4)), at a concentration (0.5-5 mg/ml) showing no inhibition of cell proliferation, stimulated insulin release from RINm5F rat insulinoma cells in a concentration- and time-related manner. But poly(2-methacryloyloxyethyl phosphorylcholine) and other synthetic phospholipid-like polymers failed to stimulate insulin release.

Angiotensin I-converting enzyme inhibitory activity and insulin secretion stimulative activity of fermented fish sauce.

Angiotensin I-converting enzyme (ACE) inhibitory peptides, Ala-Pro, Lys-Pro, and Arg-Pro, were isolated from fermented fish sauce. Five other proline-containing dipeptides having weak ACE inhibitory activity were also isolated from the fermented fish sauce. Orally administered Lys-Pro showed a tendency to lower the blood pressure of spontaneously hypertensive rats. As fermented anchovy sauce also stimulated insulin secretion by cultured RINm5F insulinoma cells, the sauce may be useful as a source of biologically active substances.