Silk Fibroin Promotes the Regeneration of Pancreatic ß-Cells in the C57BL/KsJ-Leprdb/db Mouse.

Diabetes mellitus is a chronic metabolic disease, and its progression leads to serious complications. Although various novel therapeutic approaches for diabetes mellitus have developed in the last three decades, its prevalence has been rising more rapidly worldwide. Silk-related materials have been used as anti-diabetic remedies in Oriental medicine and many studies have shown the effects of silk fibroin (SF) in both in vitro and in vivo models. In our previous works, we reported that hydrolyzed SF improved the survival of HIT-T15 cells under high glucose conditions and ameliorated diabetic dyslipidemia in a mouse model. However, we could not provide a precise molecular mechanism. To further evaluate the functions of hydrolyzed SF on the pancreatic ß-cell, we investigated the effects of hydrolyzed SF on the pancreatic ß-cell proliferation and regeneration in the mouse model. Hydrolyzed SF induced the expression of the proliferating cell nuclear antigen (PCNA) and reduced the apoptotic cell population in the pancreatic islets. Hydrolyzed SF treatment not only induced the expression of transcription factors involved in the pancreatic ß-cell regeneration in RT-PCR results but also increased neurogenin3 and Neuro D protein levels in the pancreas of those in the group treated with hydrolyzed SF. In line with this, hydrolyzed SF treatment generated insulin mRNA expressing small cell colonies in the pancreas. Therefore, our results suggest that the administration of hydrolyzed SF increases the pancreatic ß-cell proliferation and regeneration in C57BL/KsJ-Leprdb/db mice.

Rheological properties of a neutral polysaccharide extracted from maca (Lepidium meyenii Walp.) roots with prebiotic and anti-inflammatory activities.

The rheological, prebiotic, and anti-inflammatory properties of neutral polysaccharide extracted from maca roots (MP) were investigated. MP was composed of arabinose, galactose, rhamnose, and glucose. In steady shear rheological properties, an increasing concentration of MP solution exhibited higher apparent viscosity (ηa,100) and consistency index (K) under acidic condition (pH 4). In dynamic rheological properties, the dynamic moduli (G' and G″) in the frequency sweep test from the MP solution were increased with increasing concentration and decreasing pH. The changes in dynamic moduli of the MP solution with various concentrations and pH values were stable during 1 h storage at 4 °C due to the enhancement of hydrogen bonding. According to the results of the temperature sweep test, an increasing concentration of MP solution increased dynamic moduli under acidic conditions. The prebiotic properties of MP induced a higher growth of Bifidobacterium longum ATCC15707 and Lactobacillus rhamnosus ATCC 7469 than inulin and increased acetic acid, propionic acid, and butyric acid more than inulin in vitro. Furthermore, MP inhibited lipopolysaccharide-induced NO production in RAW 264.7 cells, which indicated that an increasing concentration of MP enhanced anti-inflammatory activities. Therefore, MP is a potential functional material for the food and pharmaceutical industries.

Microencapsulation of a maca leaf polyphenol extract in mixture of maltodextrin and neutral polysaccharides extracted from maca roots.

The aim of the present study was to use microencapsulation technologies to create microcapsules containing polyphenol extracted from maca leaves, which were coated with maltodextrin (MD) and neutral polysaccharides extracted from maca roots (NPMR). NPMR was composed of arabinose, galactose, rhamnose, and glucose, with the main linkage types of →5)-α-L-Araf-(1→, →2,5)-α-L-Araf-(1→, α-L-Araf-(1→, →3)-ß-D-Galp-(1→, and ß-D-Rhap-(1→. The microencapsulation efficiency of the powdered microcapsules increased with an increasing MP concentration in the coating materials. SEM images showed that, according to the increase in the MP concentration, the powdered microcapsules were more spherical and smoother, with a smaller particle size. The polyphenols extracted from the maca leaves were successfully microencapsulated in an MD-NPMR coat, as confirmed by FT-IR spectra analysis. The storage stability was greater at several temperatures for the powdered microcapsules coated with MD-MP than for those of the microcapsules coated with only MD. In an in vitro study, the powdered microcapsules coated with MD-NPMR released a smaller amount of polyphenol than the microcapsules coated with only MD in simulated gastric and intestinal fluids. Furthermore, the powdered microcapsules coated with MD-NPMR produced greater Bifidobacterium longum probiotic growth than did the microcapsules coated only with MD.

Physicochemical and sensory properties of milk supplemented with lactase microcapsules coated with enteric coating materials.

In this paper, we report the physicochemical and sensory properties of milk supplemented with a powder of microencapsulated lactase. The core material was lactase (ß-galactosidase), the primary coating material was medium-chain triglyceride (MCT), and the secondary (enteric) coating material was either hydroxypropyl methylcellulose phthalate (HPMCP) or shellac, comparing both against market milk as a control. The physicochemical properties of both types of microcapsules were analyzed, including the particle size, zeta potential, and in vitro release behavior. To survey the stability of the microcapsules in milk during storage, we studied the residual lactose content and pH. Furthermore, to determine the properties of milk supplemented with the microcapsules, changes in color and sensory properties were evaluated during storage. The particle sizes (volume-weighted mean; D[4,3]) of the microcapsules coated with HPMCP or shellac were 2,836 and 7,834 nm, respectively, and the zeta potential of the capsules coated with shellac was higher than the zeta potential of those coated with HPMCP. The pH levels of milk supplemented with the lactase microcapsules were similar to those of the control (unsupplemented market milk); however, for milk supplemented with HPMCP-coated microcapsules, the pH was slightly lower. The core material, lactase, was released from the microcapsules during 12-d storage, and 18.82 and 35.09% of lactose was hydrolyzed in the samples for HPMCP- and shellac-coated microcapsules, respectively. The sensory characteristics of milk containing microcapsules coated with HPMCP did not show significant differences from the control, in terms of sweetness or off-taste, until 8 d of storage. However, shellac-coated microcapsules showed significant difference in sweetness and off-taste at d 8 and 6 of storage, respectively. The color of milk containing HPMCP-coated microcapsules did not show a significant difference during storage. However, that containing shellac-coated microcapsules was somewhat higher in color values than others. In particular, it showed significance from 0 to 4 d storage in L* and C* values. In conclusion, a powder of lactase microcapsules coated with HPMCP can be suitable as a supplement for milk.

Butyrate attenuated fat gain through gut microbiota modulation in db/db mice following dapagliflozin treatment.

We investigated the effect of a combination treatment with dapagliflozin (Dapa), a sodium-glucose cotransporter-2 inhibitor and butyrate on weight change in db/db mice. Six-week-old male db/db mice were assigned to four groups: vehicle with normal chow diet (NCD), Dapa with NCD, vehicle with 5% sodium butyrate-supplemented NCD (NaB), or Dapa with 5% NaB. After six weeks of treatment, faecal microbiota composition was analysed by sequencing 16S ribosomal RNA genes. In the vehicle with NaB and Dapa + NaB groups, body weight increase was attenuated, and amount of food intake decreased compared with the vehicle with the NCD group. The Dapa + NaB group gained the least total and abdominal fat from baseline. Intestinal microbiota of this group was characterized by a decrease of the Firmicutes to Bacteroidetes ratio, a decrease of Adlercreutzia and Alistipes, as well as an increase of Streptococcus. In addition, the proportion of Adlercreutzia and Alistipes showed a positive correlation with total fat gain, whereas Streptococcus showed a negative correlation. Inferred metagenome function revealed that tryptophan metabolism was upregulated by NaB treatment. We demonstrated a synergistic effect of Dapa and NaB treatment on adiposity reduction, and this phenomenon might be related to intestinal microbiota alteration.

Structural and rheological properties of pectic polysaccharide extracted from Ulmus davidiana esterified by succinic acid.

The present study was carried out to investigate the physicochemical and structural properties of pectic polysaccharide extracted from Ulmus davidiana (UDP) and to determine the physicochemical, structural, and rheological properties of esterified UDP with succinic acid (ES-UDP). The results indicated that UDP had high amounts of galacturonic acids and various neutral sugars, such as galactose, rhamnose, and glucose. UDP was identified as a low methoxyl pectin, consisting of 1,4-linked α-d-GalpA (the main backbone chain), supported by the results of Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction, and 1D Nuclear magnetic resonance (NMR) spectroscopy. In the FT-IR and XRD, no difference was detected between UPD and ES-UDPs. However, 1H and 13C NMR spectra revealed that the new ester bonds were formed between a hydroxyl group of UDP and a carboxyl group of succinic acid during esterification. In the steady shear rheological analysis, the consistency index (K) of ES-UDP was significantly higher than that of UDP and increased significantly with increasing concentration of succinic acid. In the dynamic rheological analysis, the tan δ values of all ES-UDP solutions were significantly lower than those of the UDP solution.

Physicochemical, molecular, emulsifying and rheological characterizations of sage (Salvia splendens) seed gum.

The objective was to investigate the physicochemical, molecular, rheological, and emulsifying properties of water soluble-sage seed gum (WSG). WSG mainly comprised galacturonic acid and xylose. FTIR and NMR analyses confirmed the presence of pectic polysaccharides in WSG. Additionally, the molecular weight of WSG was higher than that of pectin standard. Compared to pectin standard solutions, WSG solutions exhibited higher shear thinning behavior and higher values of apparent viscosity (ηa,100) and consistency index (K) in steady shear measurements. According to the results of frequency sweep test, the dynamic moduli (G' and G″) for WSG solutions were increased with increasing frequency and concentration. The changes in dynamic moduli of WSG solutions as a function of aging time at 4 °C indicated that WSG could form a more rigid network than pectin standard. According to the results of temperature sweep test, the dynamic moduli of WSG solutions were higher than those of pectin standard solutions. Emulsion capacity and stability analyses indicated that WSG (58 and 56%, respectively) had better emulsifying properties than pectin standard (46 and 37%). In conclusion, compared to pectin standard, superior rheological and emulsifying properties observed in WSG might be related to higher molecular weight and protein content, respectively.

The characterization, selenylation and anti-inflammatory activity of pectic polysaccharides extracted from Ulmus pumila L.

The specific objective of this study was to investigate characterization, selenylation, and anti-inflammatory activities of pectic polysaccharides extracted from Ulmus pumila L. (PPU). Four different monosaccharides were found in PPU, including galacturonic acid, galactose, rhamnose, and glucose. FT-IR spectra indicated that pectic polysaccharides were successfully extracted from Ulmus pumila L., and were probably low methoxyl pectin. GC-MS and NMR analysis of PPU suggested the major monosaccharide of PPU was α-1,4-linked galacturonic acid with α-1,2-linked rhamnose as the backbone and glucose or galactose residues as branches at C-3 and C-4 positions of rhamnose. Selenylation of PPU was synthesized by 0.2 and 0.4% of sodium selenites. Selenized-PPU (Se-PPU) inhibited LPS-induced nitric oxide production in RAW 264.7 cells, and increasing selenium content enhanced anti-inflammatory properties of PPU. Therefore, Se-PPU can be used as a potential source of bioactive compounds for nutraceuticals and pharmaceutical applications.

The preventive effects of nanopowdered red ginseng on collagen-induced arthritic mice.

This study was carried out to investigate the efficiency of red ginseng nanopowder in preventing collagen-induced arthritis (CIA) in mice. The mice were divided into five groups: normal group (no immunisation), control (CIA), powdered red ginseng (PRG), nanopowdered red ginseng (NRG) and methotrexate (MTX). Administering MTX, PRG and NRG to arthritic mice significantly decreased spleen indexes, clinical and histological scores compared to control group. Serum analysis of NRG and MTX groups showed a reduction in the cytokines such as the levels of tumour necrosis factor alpha (TNF-α), interleukin 6 (IL-6) and interleukin 1ß (IL-1ß) in comparison to PRG group. The levels of immunoglobulin M (IgM) and immunoglobulin G1 (IgG1) in the NRG group were significantly lower than those of the PRG group. In summary, the present study indicated that NRG can be effective in preventing type II collagen-induced rheumatoid arthritis in mice.

Effects of selenylation modification on structural and antioxidant properties of pectic polysaccharides extracted from Ulmus pumila L.

The present study was carried out to determine the effects of selenylation on structural and antioxidant activities of pectic polysaccharides extracted from Ulmus pumila L. (PPU). Selenylation of PPU (Se-PPU) was synthesized by different concentrations of sodium selenites (0.2, 0.4, 0.6, 0.8, and 1.0g). Structural properties of Se-PPUs were characterized using ICP-OES, HPGPC, FE-SEM, XRD, FT-IR and NMR. After selenylation, the selenium contents and the molecular weights of PPU increased. In FT-IR analysis, a new intensive absorption band at around 667cm-1 was observed in Se-PPU. From 13C NMR spectra, the new peak at 62.41ppm was found in Se-PPU probably due to the substitution of selenyl groups. In vitro antioxidant activities of all samples were evaluated by reducing power, nitrite scavenging, superoxide dismutase like scavenging activities, hydroxyl radical scavenging activities, and DPPH radical scavenging activities. The antioxidant activities of Se-PPUs were significantly higher than those of PPU. In conclusion, Se-PPU exhibited the potential possibility of the use as an antioxidant in foods and pharmaceuticals. Furthermore, the expanded use of Se-PPU produced from Ulmus sources may be taken in account for better health improvement.

Abietic acid isolated from pine resin (Resina Pini) enhances angiogenesis in HUVECs and accelerates cutaneous wound healing in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Resin known as Resina Pini is listed in the Korean and Japanese pharmacopoeias and has been used for treating skin wounds and inflammation. Resin is composed of more than 50% abietic acid and 10% neutral substances. OBJECTIVE: In the present study, the wound-healing effects of abietic acid and the possible underlying mechanism of action were investigated in various in vitro and in vivo models. MATERIALS AND METHODS: The effects of abietic acid on tube formation and migration were measured in human umbilical vein vascular endothelial cells (HUVECs). Protein expression of mitogen-activated protein kinase (MAPK) activation was evaluated via Western blotting analysis. The wound-healing effects of abietic acid were assessed using a mouse model of cutaneous wounds. RESULTS: The results showed that abietic acid enhanced cell migration and tube formation in HUVECs. Abietic acid induced significant angiogenic potential, which is associated with upregulation of extracellular signal-regulated kinase (ERK) and p38 expression. Additionally, 0.8µM abietic acid-treated groups showed accelerated wound closure compared to the controls in a mouse model of cutaneous wounds. CONCLUSION: The current data indicate that abietic acid treatment elevated cell migration and tube formation in HUVECs by the activation of ERK and p38 MAPKs. We suggest that abietic acid can be developed as a wound-healing agent.

Momordica charantia (bitter melon) reduces plasma apolipoprotein B-100 and increases hepatic insulin receptor substrate and phosphoinositide-3 kinase interactions.

Aqueous extracts or juice from unripened fruit of Momordica charantia (bitter melon) has traditionally been used in the treatment of diabetes and its complications. Insulin resistance is characterized by significant down-regulation of hepatic insulin signalling as documented by attenuated phosphorylation of insulin receptor (IR), IR substrates 1 and 2, phosphoinositide-3 kinase, protein kinase B, and over-expression of phosphotyrosine phosphatase 1B. We recently demonstrated that bitter melon juice (BMJ) is a potent inhibitor of apoB secretion and TAG synthesis and secretion in human hepatoma cells, HepG2, that may be involved in plasma lipid- and VLDL-lowering effects observed in animal studies. The aim of this study was to evaluate the effects of BMJ on plasma apoB levels and hepatic insulin signalling cascade in mice fed high-fat diet (HFD). Female C57BL/6 mice (4-6 weeks old) were randomized into three groups receiving regular rodent chow, HFD and HFD+BMJ. The data indicate that BMJ not only improves glucose and insulin tolerance but also lowers plasma apoB-100 and apoB-48 in HFD-fed mice as well as modulates the phosphorylation status of IR and its downstream signalling molecules. Investigating the biochemical and molecular mechanisms involved in amelioration of diabetic dyslipidaemia by BMJ may lead to identification of new molecular targets for dietary/alternative therapies.

Possible link between NO concentrations and COX-2 expression in systems treated with soy-isoflavones.

The production of nitric oxide (NO) emerges as an essential determinant in auto- and paracrine signaling. NO is known to be generated under inflammatory conditions, carcinogenesis, and circulatory shock. The large amount of NO produced in response to cytokines plays an important role in inflammatory conditions. Cyclooxygenase (COX), the central enzyme in prostanoid biosynthesis, is involved in the first step of prostanoid synthesis from arachidonic acid. The reported studies to evaluate the relationship between NO and COX-2 have revealed both inhibitory and stimulatory effects of NO on COX-2 expression. Genistein, one of soy-isoflavones, is a polyphenolic flavonoid and a potent antioxidant and anti-inflammatory agent. In the present article, the effect of soy-isoflavones on NO production and COX-2 gene expression was examined. NO production by soy-isoflavones was greatly increased even though eNOS and iNOS expression were not different from nontreated control. The increment of NO was accompanied with the elevated expression of COX-2 and the concentrations of PGE2. The COX-2 stimulatory effect of soy-isoflavones appeared to be modulated by ERK-1 and -2 and p38. In mammalian cancer system, incubation with the NO donor sodium nitroprusside (SNP) resulted in a slight upregulation of COX-2, and cotreatment with genistein decreased COX-2 expression possibly by the activation of AMP-activated protein kinase (AMPK).