Changes in composition and functional properties of proteins and their contributions to Nham characteristics

Changes in composition and functional properties of proteins during fermentation of Nham, a Thai-fermented sausage, were studied. An alkaline-soluble fraction constituted a major protein component of Nham. The amount of each protein fraction in Nham varied, depending on the fermentation time. As fermentation proceeded, the progressive decrease in sarcoplasmic and myofibrillar protein fractions was accompanied by an increase in the alkaline-soluble fraction and non-protein constituents (P<0.05). Slow pH lowering to pH 4.6 during fermentation as a result of bacterial growth and accumulation of lactic acid affected the molecular conformation of the muscle proteins and resulted in changes in protein functional properties. The acid produced resulted in changes in solubility, water-binding capacity, textural properties, and color characteristics. Proteolysis of Nham proteins occurred during fermentation, resulting in increases in TCA-soluble peptides and free α-amino acids, which may contribute to the taste and aroma of Nham.     

Skin improvement and stability of Echinacea purpurea dermatological formulations

Echinacea purpurea contains many beneficial constituents for protection of skin from oxidative stress and for improving hydration of skin. This study aimed to investigate the stability and dermatological efficacy of E. purpurea cream and gel. Echinacea purpurea extract was incorporated into suitable cream and gel bases. Stability of the extract in the formulations was investigated by determining its residual total phenolic content and antioxidant activity after storage at 4°C, 30°C and 40°C for 6 months. The effect of those formulations on skin irritation, hydration level and wrinkle reduction was evaluated in 10 healthy volunteers, aged 25–40 years. The shelf lives of E. purpurea cream and gel in terms of total phenolic content and antioxidant activity were only 2 and 4 months respectively at 4°C and could be extended up to 7 months by incorporation of α‐tocopherol or disodium editate. The corneometer hydration indices increased up to 10.6 AU and 11.4 AU, and the wrinkles decreased 9.47% and 14.92% because of the application of E. purpurea cream and gel for 1 month. Both formulations showed no irritation to skin. Echinacea purpurea cream and gel developed in this study were effective in improving skin hydration and reducing wrinkle, but showed low storage stability.     

Poly(styrene)‐ and Poly(styrene‐co‐methyl methacrylate)‐graft‐hydrogenated natural rubber latex: Aspect on synthesis,properties, and compatibility

The undesirable properties of natural rubber (NR) can be improved via hydrogenation and graft copolymerization. Hydrogenated NR (HNR) latex was prepared via diimide reduction and then grafted with styrene (ST) or ST/methyl methacrylate (MMA) to form poly(ST)‐graft‐HNR (poly(ST)‐g‐HNR, GHNRS) or poly(ST‐co‐MMA)‐g‐HNR (GHNRSM), respectively. For the grafting of ST monomer onto HNR particles, the %monomer conversion and %grafting efficiency (%GE) were monitored as functions of %hydrogenation, monomer and initiator concentrations, temperature, and time. Under the optimum condition (HNR with 54.3% hydrogenation; 100 phr of ST, 1 phr of initiator at 50°C for 8 h), maximum %conversion and %GE of 44.6% and 36.9%, respectively, were achieved. Thermogravimetric analysis revealed that the HNR grafted with ST or ST/MMA had higher decomposition temperature than an ungrafted one. When these graft products were blended at 10% (w/w) with acrylonitrile‐butadiene‐styrene (ABS) resin, the GHNRS/ABS and GHNRSM/ABS composites exhibited the higher flexural strength and heat aging tolerance compared to the ungrafted HNR/ABS composite. Scanning electron microscopy (SEM) also showed the higher degree of homogeneity at the fractural surface, supporting the higher compatibility between the ABS and the GHNRS or GHNRSM phases in the blends. J. VINYL ADDIT. TECHNOL., 22:100–109, 2016. © 2014 Society of Plastics Engineers     

Structural analysis of lead fullerene-based inhibitor bound to human immunodeficiency virus type 1 protease in solution from molecular dynamics simulations

Molecular dynamics (MD) simulations of the HIV-1 protease (HIVP) complexed with lead fullerene-based inhibitor (diphenyl C60 alcohol) in the three protonated states, unprotonated (Un-), monoprotonated (Mono-), and diprotonated (Di-) states at Asp25 and Asp25' were performed. As the X-ray structure of the investigated complex is not available, it was built up starting with the X-ray crystallographic structure of the HIVP complexed with non-peptide inhibitor (PDB code: 1AID) and that of the diphenyl C60 alcohol optimized using the integrated ONIOM molecular orbital calculations. The inhibitor was, then, introduced into the enzyme pocket using a molecular docking method. Change of the HIVP binding cavity for all three states were evaluated in terms of distance between the two catalytic residues, Asp25 and Asp25' as well as those between the catalytic residues and the flap regions. The torsional angles formed by the O-C-C-O of the two carboxyl groups of the catalytic dyad show the non-planar configuration with the most frequency at about -45 degrees for the Un-, 35 degrees and -95 degrees for the Mono- and 60 degrees for the Di-systems. At equilibrium, different orientations of the fullerene-based inhibitor in the three protonation states were observed. For the Di-state, the OH group of the inhibitor stably forms hydrogen bonds with the two aspartic residues. It turns to the flap region to form hydrogen bonding to the backbone N of Ile50' for the Un-state. In contrast, the OH group turns to locate between the catalytic and the flap region for the Mono-states. Beside the molecular orientation, the rotation of the OH group of the inhibitor in the Un-state was also detected. In terms of solvation, the carboxylate oxygens of the aspartic residues in the Un- and Mono-states were solvated by one to three water molecules while the OH group in these two states was coordinated by one water molecule. This is in contrast to the Di-state in which no water molecule is available in the radius of 5-6A around the oxygen atoms of the carboxylate groups of enzyme and of the OH group of the inhibitor. The simulated results lead to the conclusion that the active site of the HIVP complexed with the diphenyl C60 alcohol is the diprotonation states on Asp25 and Asp25'.     

Heat-induced formulation inhomogeneity of a three-component suspension

A suspension formulation containing sarafloxacin HCl, triamcinolone acetonide, and clotrimazole was developed for the treatment of otitis externa in dogs. The potency for the three active ingredients in this suspension was monitored at 25°C and 40°C for up to 3 months. The potencies of triamcinolone and clotrimazole were found unchanged, but the potency of sarafloxacin HCl in the samples stored at 40°C for 1 month varied significantly between samples. However, assay inconsistency for sarafloxacin HCl was not seen in samples stored at 25°C. Under an optical microscope, large crystals were found in the 40°C stability samples but not in the 25°C samples. The large crystals in 40°C samples were identified as sarafloxacin by high-performance liquid chromatography (HPLC). This finding suggests that crystal growth of sarafloxacin took place at 40°C during storage, leading to the formation of larger crystals and the consequent sampling nonuniformity and assay inconsistency. The solid-state properties of these crystals were further evaluated using hot-stage microscopy and Fourier-transform infrared (FTIR) analysis. The results indicate that the crystal growth of sarafloxacin was most likely attributed to a change in the hydration form of sarafloxacin.     

Thermal Behavior of Ursodeoxycholic Acid in Urea: Identification of Anomalous Peak in the Thermal Analysis

The objective of this study was to clarify the thermal behavior of ursodeoxycholic acid (UDCA) in mixtures with urea. Physical mixtures of UDCA and urea in various ratios were prepared, and the thermal analysis of these sample mixtures was investigated using conventional differential scanning calorimetry (DSC) and variable-temperature powder X-ray diffractometry (VTXRD). The hot-stage microscopy (HSM) and powder X-ray diffractometry (PXRD) were used as complementary techniques. From the DSC results of all sample mixtures, it was found that there was no endothermic peak at the melting temperature of intact UDCA crystals. The DSC thermograms of each ratio showed only the endothermic peak at about 136°C due to the melt of urea and the anomalous endothermic peak at about 155°C-157°C. The VTXRD study revealed that the crystals of urea completely disappeared at a temperature of 140°C. At this temperature, it was identified that the VTXRD pattern obtained was of UDCA crystals. The crystalline peaks gradually decreased in intensity at a temperature of 150°C. When the temperature was up to 160°C, the identical crystalline peaks of UDCA crystals completely disappeared. It was concluded that the anomalous endothermic peak at 155°C-157°C was the peak due to the dissolution of UDCA crystals in the surrounding melted urea.     

Efficacy of chitin-PAA-GTMAC gel in promoting wound healing: animal study

Acrylic grafted chitin (chitin-PAA) was modified with glycidyltrimethylammonium chloride (GTMAC) with the aim of promoting wound healing. The chitin-PAA-GTMAC gels with different GTMAC contents were compared with the original chitin-PAA gel and Intrasite gel for their efficacy in deep wound healing of Wistar rats. Four full-thickness wounds were made on the dorsal skin of rats and then each was treated with 4 materials; chitin-PAA, chitin-PAA-GTMAC(1:4), chitin-PAA-GTMAC(1:10) and Intrasite gel. During 18 days of treatment, the wounds were visually observed and calculated for wound size using image analysis program. Skin wound tissues of sacrificed rats were processed for routine histological observation and immunohistochemistry of proliferating cell nuclear antigen (PCNA). The wounds covered with the chitin derivatives either with or without GTMAC showed a significant reduction in wound size in day 9 in comparison with day 12 for those covered with Intrasite gel. The faster rate and the better pattern of epidermal development observed in histological study as well as the higher dermal cell proliferation (PCNA expression) also demonstrated the better efficiency in wound healing of the chitin derivatives than Intrasite. The earliest epidermal development of the wounds treated with chitin-PAA-GTMAC (1:4) among the tested materials suggested the most promising of this material for the treatment of full-thickness open wound.     

Changes in lipid composition and fatty acid profile of Nham,a Thai fermented pork sausage,during fermentation

Changes in lipid composition and fatty acid profile of Nham during fermentation were investigated. Total lipids of Nham were in the range 2–3%. The extracted lipid of initial Nham mix consisted mainly of triglycerides (TG), accounting for more than 75% of the total lipid, followed by phospholipids (PL) and a trace amount of diglycerides (DG) and free fatty acid (FFA). During fermentation, TG, DG and PL decreased with a concomitant increase in FFA, indicating lipolysis of Nham lipids during fermentation. Changes in fatty acids of the total lipids, non-polar and polar lipid fractions were observed during fermentation. In both total and non-polar lipid fractions, the major fatty acids found in a descending order were oleic (C18:1), linoleic (C18:2) and palmitic (C16:0) acids, which together accounted for 90% of the total fatty acids. Increases in fatty acid contents in both total and non-polar lipid fractions, were observed with a corresponding decrease in the quantity of fatty acids of phospholipids. As the fermentation proceeded, peroxide value generally increased while TBARS values decreased. Overall, lipid oxidation in Nham occurred during fermentation but did not cause the objectionable odour and taste in any Nham tested.     

FLT3-specific curcumin micelles enhance activity of curcumin on FLT3-ITD overexpressing MV4-11 leukemic cells

Curcumin, a major active compound in the turmeric rhizome, has many biological properties, especially anti-leukemia activity. The overexpression of FMS-like tyrosine kinase 3 protein with internal tandem duplication (FLT3-ITD) mutation protein was related to the poor prognosis and disease progression of leukemia. In this study, the cytotoxicity and inhibitory effect of curcumin on cell cycle of FLT3-ITD overexpressing MV4-11 leukemic cells were evaluated. Moreover, curcumin polymeric micelles conjugated with FLT3-specific peptide (FLT3-Cur-micelles) were prepared using a film hydration method to increase curcumin solubility and the inhibitory effect on MV4-11 cells was evaluated. Cytotoxicity and cell cycle analysis were performed using an MTT assay and flow cytometry, respectively. Physical properties of FLT3-Cur-micelles, including particle size, size distribution, morphology, and entrapment efficiency (EE), were evaluated. Cellular uptake of the micelles on MV4-11 cells was determined by flow cytometry and fluorescence microscopy. FLT3-Cur-micelles were observed with size less than 50?nm and high EE of >75%. In addition, FLT3-Cur-micelles demonstrated excellent internalization and increased curcumin accumulation in leukemic cells when compared to free curcumin. Furthermore, FLT3-Cur-micelles exhibited a strong cytotoxic effect on MV4-11 cells with IC₅₀ value of 1.1?µM, whereas the blank micelles showed no effect. Furthermore, FLT3-Cur-micelles showed no significant effect on normal human PBMCs with IC₅₀ value >25?µM. In summary, FLT3-Cur-micelles are a promising nanocarrier system for enhancing anti-leukemic activity of curcumin and suitable for further preclinical studies.