HP Content Research Articles

Quality assessment method for Chinpi, dried Citrus spp. peel and its derived Kampo medicines using specific monoclonal antibody against hesperidin.

Hesperidin (hesperetin 7-rutinoside, HP), a flavonoid glycoside found in Citrus unshiu Marcowicz or Citrus reticulata Blanco (Rutaceae), has been reported to exert a variety of pharmacological effects. As the efficacies and qualities of their dried peel, Chinpi and its derived Kampo medicines can be evaluated by their HP contents, a method for HP detection must be developed. To produce a specific monoclonal antibody against HP (mAb 5D12) to detect the HP contents in Japanese traditional medicines via indirect competitive enzyme-linked immunosorbent assay (icELISA). BALB/c mice were immunised with many haptens of HP-bovine serum albumin (BSA) conjugates that were prepared using sodium periodate (NaIO4 ) to cause an immune response. In addition, conventional hybridoma techniques were utilised to generate mAb 5D12. The detection range of HP by the mAb 5D12-based icELISA was 1.56-25.0 ng/mL, with a detection limit of 1.12 ng/mL. The maximum coefficient of variation, as evaluated from the intra- and inter-assays, was <10.0%, and the percentages of recovery, as determined by the spike-recovery tests, were 105%-115%. Moreover, the HP content, which was obtained from the developed icELISA, correlated well with that obtained via high-performance liquid chromatography-ultraviolet (HPLC-UV). These validation analyses revealed that the established icELISA technique exhibited high precision and accuracy. Notably, this is the first report on the development of icELISA for the HP content-based quality control of Chinpi and its derived Kampo medicines.

A novel emulsion gel solely stabilized by the hot water extracted polysaccharide from psyllium husk: Self-healing plays a key role

The hot water extracted polysaccharide (HP) from psyllium husk has been proved to have unique gelling properties. The present work was first to focus on the self-healing ability of the HP hydrogel. Rheological and macroscopic tests profiled the fast recoverable property of the HP gel network. Then, for the first time, we explored the role of self-healing ability in the fabrication of emulsion gels. The effect of HP concentration on the stability and physical properties of the HP emulsion gel were investigated from the aspects of particle size, macroscopic state, microscopic structure, and rheological properties. The results showed that the emulsion could be well stabilized only by HP. And when HP content reached the gel concentration, the stability remarkably increased. Further, the underlying mechanism was illustrated. It seemed that the interfacial HP interacted with adjacent polysaccharide molecules located on the interface or in the continuous phase to entangle a whole network structure. Consequently, those oil droplets were well immobilized in the network structure. In summary, a stable emulsion gel was simply and feasibly fabricated based on the self-healing property of HP, which is of great potential in food and biomedical applications. • Hot water extracted polysaccharides(HP) from psyllium husk had a good self-healing ability. • The self-healing ability of HP gel played a key role in the fabrication of emulsion gels. • The influence of HP concentration on the stability of emulsion gel was investigated. • The stabilization mechanism of HP emulsion gels was proposed.

Oral Administration of Acrylamide Worsens the Inflammatory Responses in the Airways of Asthmatic Mice Through Agitation of Oxidative Stress in the Lungs.

Acrylamide is a toxic chemical substance produced when starch-rich foods are fried at high temperatures. Asthma is a chronic and complicated respiratory disease, of which genetic and environmental factors are the main triggers. Orally-received components may have an effect on asthma pathophysiology. The aim of this study was to investigate the role of AA as a stimulus in asthma. BALB/c mice were allocated into four groups as follows: two OVA-sensitized asthmatic groups, including one treated with AA by gavage feeding and one non-treated (asthma group), and two healthy (non-asthmatic) groups, one treated with AA by gavage feeding and one non-treated (negative control group). Airway hyperresponsiveness, cell count, cytokine levels in BAL fluid, lung histopathology, IgE levels, and oxidative stress indices including plasma level of MDA, pulmonary antioxidant enzymes (SOD and CAT) levels, HP content, and collagen fiber accumulation in lung tissue were measured. We found that the group of mice treated with both OVA and AA (asthmatic and AA-treated mice) experienced higher levels of asthma-associated biomarkers, including higher enhanced pause (Penh value), eosinophilic inflammation, mucus hyper secretion, goblet cell hyperplasia, total and OVA-specific IgE levels, IL-4, IL-5, and IL-13 levels than the group sensitized only with OVA (asthmatic mice). The OVA-AA-treated mice also experienced worsened levels of oxidative stress indicators. Healthy (non-asthmatic) mice that only received AA were in similar conditions to healthy untreated mice (negative control group). The OVA-AA-treated group showed more severe allergic asthma symptoms in comparison to the group only sensitized with OVA. Therefore, food/water contaminated with AA can act as a stimulant of allergic asthma and exacerbate the bronchial inflammatory responses.

Evaluation of the Hydroxyproline Content in the Frozen Allograft Tendon Impregnated with the Mesenchymal Cells & PRP in Lamb

Background: Tendon grafting is mostly required to repair an injury to flexor tendon and due to the importance of this tendon; it needs methods to speed up the re-vitality of the tendon allograft. OBJECTIVES: The aim was to investigate the efficacy of the mesenchymal cells and platelet rich plasma on the hydroxyproline content of frozen allograft tendons after grafting in lamb. METHODS: In this experimental study, mid portion (five cm) of SDFT of fifteen lambs, from both forelimbs were removed and replaced with frozen allografts tendon. Animals were divided into 3 subgroups of 5 lambs each: control, PRP and MSCs. The hydroxyproline concentration was measured by after 60 days. Data was analyzed using student t test at P<0.05%. RESULTS: The normal mean values of hydroxyproline content was 137.171±5.291 mg/g dry matter which 87.694±6.502 in control group but 99.694±1.839 in PRP group and in group treated with MSC was recorded 134.322±2.123 mg/g dry matter (P<0.05%) it was quite significantly different between control tendons with that of normal one of the same animal. There was marked increase in hydroxyproline content of MSC group when compare with that of control and PRP groups. CONCLUSIONS: The results of this study showed that MSCs could enhance HP content in the frozen grafted tendon in lamb.

Influence of Alkali and Alkaline Earth Metallic Species on the Phenolic Species of Pyrolysis Oil

Bio-oil as an important renewable energy product has been successfully made from corn stalks and lignin via a fast pyrolysis process. This study investigated the effects of alkali and alkaline earth metallic species (AAEMs) on phenolic products in corn stalk and lignin pyrolysis oil. Corn stalks were demineralized with 0.5 M HCl, and lignin was doped with 0.2wt%, 2wt%, and 20wt% KCl and CaCl2, respectively. The pyrolysis experiments were conducted in a fixed bed tubular furnace ranging from 450 °C to 600 °C. It was found that AAEMs exert positive effects on the formation of char and gas and inhibit the production of bio-oil. The effect of KCl on the product distribution from lignin is somewhat stronger than CaCl2. Moreover, the content of P, HP, GP, and SP decreases after removal of AAEMs. KCl can promote the polymerization of light molecular aromatic hydrocarbons to increase the SP yield at 600 °C, whereas the effect on increase in P and HP content is relatively weaker than CaCl2. KCl and CaCl2 play notable roles in demethxylation and demethoxylation in increasing phenol content and removing the R group from the ring.

Amino-terminated hyperbranched polymer for toughness improvement of epoxy/clay nanocomposites

Amino-terminated hyperbranched poly(ester-amine) (HP) was prepared and used as a toughening agent for epoxy resin in presence of Montmorillonite organoclay (OMMT) as a reinforcing filler. Several parameters which affect the physico-mechanical properties of epoxy/clay nanocomposites were studied including HP content, clay concentration, and intercalation/exfoliation and degree of dispersion of OMMT clay within the epoxy matrix. The results showed a significant improvement in toughness, physico-mechanical properties and enhancement in thermal stability of epoxy/clay nanocomposites upon using HP (20 wt%) as a toughening agent at low OMMT clay loading (1 wt%). These promising results could open the way for epoxy resins to be used in more high-impact resistance applications.

Reishi Mushroom Attenuates Hepatic Inflammation and Fibrosis Induced by Irradiation Enhanced Carbon Tetrachloride in Rat Model

This work was undertaken to establish a new experimental model of hepatic fibrosis by gamma irradiation and CCl4 and to study the hepatoprotective effect of Reishi Mushroom (RM) against hepatic fibrosis induced in that model. Our results revealed that oral co-administration of 110 mg/kg RM by gavage to fibrotic rats offered an obvious hepatic protection as assured by the significant decrement in ALT and AST, HP content, MDA and NO levels with elevation of the antioxidant enzymes activities. The levels of TGF-β, TNF-α, HO-1 and type-1 collagen and their m-RNA expression were markedly declined as compared with those of fibrotic rats. Microscopical examination revealed that the exposure of rats to radiation aggravated the effect of CCl4 causing extensive collagen deposition and marked pseudolobulation of the hepatic parenchyma indicative of bridging fibrosis. While, oral co-administration of RM obviously improved the state of steatosis and apparently suppressed hepatic fibrogenesis.

Improved Osteochondral Allograft Preservation Using Serum-Free Media at Body Temperature

Background: Osteochondral allografts (OCAs) are currently preserved at 4°C and used within 28 days of donor harvest. The window of opportunity for implantation is limited to 14 days due to a 2-week disease testing protocol. Hypothesis: Osteochondral allograft tissues stored at 37°C will have significantly higher chondrocyte viability, as well as superior biochemical and biomechanical properties, than those stored at 4°C. Study Design: Controlled laboratory study. Methods: Osteochondral allografts from 15 adult canine cadavers were aseptically harvested within 4 hours of death. Medial and lateral femoral condyles were stored in Media 1, similar to the current standard, or Media 2, an anti-inflammatory and chondrogenic media containing dexamethasone and transforming growth factor−β3, at 4°C or 37°C for up to 56 days. Chondrocyte viability, glycosaminoglycan (GAG) and collagen (hydroxyproline [HP]) content, biomechanical properties, and collagen II and aggrecan content were assessed at days 28 and 56. Five femoral condyles were stored overnight and assessed the next day to serve as controls. Results: Storage in Media 1 at 37°C maintained chondrocyte viability at significantly higher levels than in any other media-temperature combination and at levels not significantly different from controls. Osteochondral allografts stored in either media at 4°C showed a significant decrease in chondrocyte viability throughout storage. Glycosaminoglycan and HP content were maintained through 56 days of storage in OCAs in Media 1 at 37°C. There were no significant differences in elastic or dynamic moduli among groups at day 56. Qualitative immunohistochemistry demonstrated the presence of collagen II and aggrecan throughout all layers of cartilage. Conclusion: Osteochondral allograft viability, matrix content and composition, and biomechanical properties were maintained at “fresh” levels through 56 days of storage in Media 1 at 37°C. Osteochondral allografts stored at 4°C were unable to maintain viability or matrix integrity through 28 days of storage. These findings suggest that storage of OCAs in a defined media at 37°C is superior to current protocols (4°C) for tissue preservation prior to transplantation. Clinical Relevance: Storage of OCAs in serum-free chemically defined media at 37°C can increase the “window of opportunity” for implantation of optimal tissue from 14 days to 42 days after disease testing clearance.

The effect of hyperbaric oxygen treatment on aspiration pneumonia

We have studied whether hyperbaric oxygen (HBO) prevents different pulmonary aspiration materials-induced lung injury in rats. The experiments were designed in 60 Sprague-Dawley rats, ranging in weight from 250 to 300g, randomly allotted into one of six groups (n=10): saline control, Biosorb Energy Plus (BIO), hydrochloric acid (HCl), saline+HBO treated, BIO+HBO treated, and HCl+HBO treated. Saline, BIO, HCl were injected into the lungs in a volume of 2ml/kg. A total of seven HBO sessions were performed at 2,4atm 100% oxygen for 90min at 6-h intervals. Seven days later, rats were sacrificed, and both lungs in all groups were examined biochemically and histopathologically. Our findings show that HBO inhibits the inflammatory response reducing significantly (P<0.05) peribronchial inflammatory cell infiltration, alveolar septal infiltration, alveolar edema, alveolar exudate, alveolar histiocytes, interstitial fibrosis, granuloma, and necrosis formation in different pulmonary aspiration models. Pulmonar aspiration significantly increased the tissue HP content, malondialdehyde (MDA) levels and decreased (P<0.05) the antioxidant enzyme (SOD, GSH-Px) activities. HBO treatment significantly (P<0.05) decreased the elevated tissue HP content, and MDA levels and prevented inhibition of SOD, and GSH-Px (P<0.05) enzymes in the tissues. Furthermore, there is a significant reduction in the activity of inducible nitric oxide synthase, TUNEL and arise in the expression of surfactant protein D in lung tissue of different pulmonary aspiration models with HBO therapy. It was concluded that HBO treatment might be beneficial in lung injury, therefore, shows potential for clinical use.

Synthesis and characterization of Poloxamer 188-grafted heparin copolymer

Background: Poloxamer 188 is a safe biocompatible polymer that can be used in protein drug delivery system. Aim: In this study, a new heparin–poloxamer 188 conjugate (HP) was synthesized and its physicochemical properties were investigated. HP structure was confirmed by Fourier transform infrared spectroscopy (FTIR) and Hydrogen-1 nuclear magnetic resonance spectroscopy (1H-NMR). Content of the conjugated heparin was analyzed using Toluidine Blue. The critical micelle concentration (CMC) of the copolymer was determined by a fluorescence probe technique. The effect of HP on the gelation of poloxamer 188 was characterized by the rheological properties of the HP–poloxamer hydrogels. Solubility and viscosity of HP were also evaluated compared with poloxamer 188. Results: From the results, the solubility of the conjugated heparin was increased compared with free heparin. The content of heparin in HP copolymer was 62.9%. The CMC of HP and poloxamer 188 were 0.483 and 0.743 mg/mL, respectively. The gelation temperature of 0.4 g/mL HP was 43.5°C, whereas that of the same concentration of poloxamer 188 was 37.3°C. With HP content in poloxamer 188 solution increasing, a V-shape change of gelation temperature was observed. Conclusion: Considering the importance of poloxamer 188 in functional material, HP may prove to be a facile temperature-sensitive material for protein drug-targeted therapy.

Anastomotic healing in ileum and colon of alloxan-induced diabetic rats

To investigate if diabetes mellitus may alter breaking strength (BS) and collagen content in ileum and colon anastomoses in rats. Three-hundred Wistar rats were randomly assigned to 5 experimental groups, 60 per group: normal controls surgically manipulated (G1); normal controls submitted to ileum (G2) and colon (G3) anastomotic construction; diabetic rats submitted to ileum (G4) and colon (G5) anastomotic construction. Each group was further divided into 6 subgroups with 10 rats each for sacrifice at 0, 4, 7, 14, 21, and 30 days after surgery. All surgical procedures were performed 3 months after alloxan diabetes induction. BS was measured in all intestinal anastomoses. Fragments of ileum and colon anastomoses were taken for hydroxyproline concentration (HP) and total tissue protein (TP) dosages. Anastomotic BS was significantly decreased (P<0.05) in ileum and colon of G4 and G5 diabetic groups up to 7 and 14 days after surgery, respectively, compared with G2 and G3 normal control groups. Anastomotic HP and TP content did not significantly differ between diabetic and normal control operated groups in ileum or colon at all evaluation times. Experimental diabetes leads to impaired intestinal anastomotic strength during early surgical wound repair, but does not appear to be implicated with collagen synthesis capacity.

Self-association properties of HPMA copolymers containing an amphipathic heptapeptide

Receptor-binding peptides are suitable targeting moieties for macromolecular therapeutics. Binding several targeting peptides to one macromolecule may improve biorecognition due to the multivalency effect. On the other hand, the resulting amphipathic structure of such conjugates may result in the association of side-chains with a concomitant decrease in the accessibility of the side-chain-bound ligands. Using the one-bead one-compound combinatorial method, we have recently identified a heptapeptide (YILIHRN; HP) ligand for the CD21 receptor (Biomacromolecules 7, 3037, 2006). Here, we evaluated the relationship between structure and self-association of N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-HP conjugates using fluorescence resonance energy transfer (FRET) to evaluate their conformation in solution. In addition to HP, HPMA copolymers containing side-chains terminating in tryptophan (energy donor) and dansyl (energy acceptor) were synthesized, and solutions were evaluated using an excitation wavelength of 295 nm (ratio of emission intensity 510 nm/370 nm indicated energy transfer efficiency). It was found that higher HP content correlated with higher FRET efficiency, indicating the formation of compact coils. Modification of the HPMA copolymer backbone by the incorporation of acrylic acid (AA) comonomer units resulted in decreased FRET efficiency, presumably due to the expansion of the polymer coils as a result of electrostatic repulsion. The dependence of FRET efficiency on pH was in agreement with the ionization profile of the AA residues.To determine the effect of HP content on enzymatic drug release kinetics, HPMA copolymer-HP conjugates containing GFLG side-chains terminating with doxorubicin (DOX) were incubated with papain and the release of free DOX monitored. When HP content increased above a particular threshold, the rate of DOX release decreased as a result of self-association of HPMA copolymer-GFLG-DOX-HP conjugates. The FRET data correlated well with hydrodynamic volumes determined by size exclusion chromatography (SEC), with molecular weights determined by light scattering, and with the kinetics of drug release.

Influence of hydrophobe content on the solution rheology of hydrophobically modified terpolymer of SO 2, N, N-diallyl- N-carboethoxymethylammonium chloride

Rheological properties of hydrophobically modified copolymer of SO 2, N, N-diallyl- N-carboethoxymethylammonium chloride and the hydrophobic monomer N, N-diallyl- N-octadecylammonium chloride were studied. The influence of hydrophobe content (HP) and polymer concentration was investigated. Polymers with HP content in the range 1.5–5% were examined and the concentration was varied in the range 2–5 wt%. Both dynamic and steady-shear experiments were performed in ARES rheometer. Copolymers were observed to exhibit typical viscoelastic behavior even with low HP content. Both the dynamic viscosity, η′ and storage modulus, G′, increase with the increase of both the polymer concentration and the HP content of the system. The viscosity of the high HP content polymer showed a strong shear dependency, while G′ was a weak function of frequency and gel-like behavior was observed. The zero-shear viscosity, η 0, showed a strong concentration dependency ( η 0 ∼ ϕ α ; 1.1 < α < 5.9). The concentration dependency of η 0 suggests that intermolecular association is dominant in the high HP content polymer. Control of the HP content and polymer concentration of this class of polymers can lead to a wide range of interesting rheological properties.

Synthesis of porphyrin-introduced silica gels by sol–gel process

Using a hydroxyl group appended free base porphyrin derivative (HP), porphyrin-introduced silica gels were synthesized by a sol–gel process. The HP content in the materials linearly increased with increasing the HP concentration. Meanwhile, free base tetraphenylporphyrin (TPP) with no hydroxyl groups were almost not incorporated into the silica gels. These facts suggested that the interaction between hydroxyl groups of the HP molecules and silica network is considerably strong. The UV–vis characters of HP-introduced materials were almost the same as pure HP molecules. The Beer's plot indicated that the HP molecules in the materials are dispersed.

Constituent and sensory changes of different types of carrots after freezing and frozen storage

The constituents of 6 different strains of carrots were conducted and evaluated of different types for suitability in the freezing processing. The Okinawa strain before freezing was the calcium and low pectin. Since the freeze processing result in a reduction in calcium and pectin, this type is not suitable for freezing. The Chantenay strain and Long Orange strain before freezing had pectin contents. There showed little changes in ratio of WP, PP and HP content freezing and frozen storage, making this type suitable for freezing. The Danvers strain had calcium and pectin contents were also high. There were few changes due to freezing and frozen storage, was rated highly in terms of its texture, making this type suitable for freezing. The Imperator strain was little change in quantity of calcium and pectin. Moreover, the amount of WP was low, and was rated highly for texture after freezing and frozen storage. making this strain suitable for freezing. The Sanzun strain had low calcium content and high pectin content. There were negligible changes due to freezing, thus making it suitable for freeze processing.

Estimation of creatine phosphokinase and hydroxyproline in the developing limb: its use in evaluating the effect of teratogens on myogenesis and chondrogenesis.

Forelimbs of mouse fetuses were examined for tissue-specific, drug-induced alterations in their biochemical composition. The activity of the enzyme creatine phospholinase (CPK; to estimate myogenesis) and the content of hydroxyproline (HP; to estimate chondrogenesis) were compared in homogenates of control and treated mouse-fetus forelimbs on day 14 of gestation. In addition, the content of DNA, RNA, and protein was also measured. Injection of 6-aminonicotinamide (6-AN) (15 mg/kg) on day 10 resulted in an overall growth retardation in day 14 fetuses and all biochemical parameters tested were reduced. The ratio of PH:CPK was unaffected by 6-AN treatment. Retinoic acid (vitamin A acid; 100 mg/kg), administered to pregnant female mice on day 10, produced severe forelimb defects and resulted in a signific 10 resulted in an overall growth retardation in day 14 fetuses and all biochemical parameters tested were reduced. The ratio of PH:CPK was unaffected by 6-AN treatment. Retinoic acid (vitamin A acid; 100 mg/kg), administered to pregnant female mice on day 10, produced severe forelimb defects and resulted in a signific 10 resulted in an overall growth retardation in day 14 fetuses and all biochemical parameters tested were reduced. The ratio of PH:CPK was unaffected by 6-AN treatment. Retinoic acid (vitamin A acid; 100 mg/kg), administered to pregnant female mice on day 10, produced severe forelimb defects and resulted in a significant reduction in day 14 forelimb HP and RNA content, without altering CPK, DNA, or protein; thus, the HP:CPK ratio was decreased. These results indicated that 1) 6-AN nonspecifically retards growth and cyto-differentiation in limbs; 2) retinoic acid inhibits synthesis of collagen and RNA; 3) retinoic acid has a differential effect upon chondrogenic and myogenic tissues of the limb, and 4) the comparison of HP content and CPK activity in tissue homogenates is an acceptable method of evaluating teratogenic compounds for selective effects.