Nitrogen-15 dynamic nuclear polarization of nicotinamide derivatives in biocompatible solutions.

Dissolution dynamic nuclear polarization (dDNP) increases the sensitivity of magnetic resonance imaging by more than 10,000 times, enabling in vivo metabolic imaging to be performed noninvasively in real time. Here, we are developing a group of dDNP polarized tracers based on nicotinamide (NAM). We synthesized 1-15N-NAM and 1-15N nicotinic acid and hyperpolarized them with dDNP, reaching (13.0 ± 1.9)% 15N polarization. We found that the lifetime of hyperpolarized 1-15N-NAM is strongly field- and pH-dependent, with T1 being as long as 41 s at a pH of 12 and 1 T while as short as a few seconds at neutral pH and fields below 1 T. The remarkably short 1-15N lifetime at low magnetic fields and neutral pH drove us to establish a unique pH neutralization procedure. Using 15N dDNP and an inexpensive rodent imaging probe designed in-house, we acquired a 15N MRI of 1-15N-NAM (previously hyperpolarized for more than an hour) in less than 1 s.

Cambogin suppresses dextran sulphate sodium-induced colitis by enhancing Treg cell stability and function.

BACKGROUND AND PURPOSE: Inflammatory bowel disease (IBD) is a chronic and relapsing inflammatory disorder of the gastrointestinal tract, and an impaired immune response plays a critical role in IBD. The current drugs and therapies for IBD treatment are of limited use, therefore, there is a need to find novel drugs or therapies for this disease. We investigated the effect of cambogin in a mouse model of dextran sulphate sodium (DSS)-induced colitis and whether cambogin attenuates inflammation via a Treg-cell-mediated effect on the immune response. EXPERIMENTAL APPROACH: Chronic colitis was established in mice using 2% DSS, and cambogin (10 mg·kg-1 , p.o.) was administered for 10 days. Body weight, colon length and colon histology were assessed. Cytokine production was measured using elisa and quantitative real-time PCR. To evaluate the mechanism of cambogin, human CD4+ CD25hi CD127lo Treg cells were isolated from peripheral blood mononuclear cells. Major signalling profiles involved in Treg cell stability were measured. KEY RESULTS: Cambogin attenuated diarrhoea, colon shortening and colon histological injury and IL-6, IFN-γ and TNF-α production in DSS-treated mice. Cambogin also up-regulated Treg cell numbers in both the spleen and mesenteric lymph nodes. Furthermore, cambogin (10 µM) prevented Foxp3 loss in human primary Treg cells in vitro, and promoted USP7-mediated Foxp3 deubiquitination and increased Foxp3 protein expression in LPS-treated cells. CONCLUSIONS AND IMPLICATIONS: The effect of cambogin on DSS-induced colitis is expedited by a Treg-cell-mediated modification of the immune response, suggesting that cambogin could be applied as a novel agent for treating colitis and other Treg cell-related diseases.

Torilin Inhibits Inflammation by Limiting TAK1-Mediated MAP Kinase and NF-κB Activation.

Torilin, a sesquiterpene isolated from the fruits of Torilis japonica, has shown antimicrobial, anticancer, and anti-inflammatory properties. However, data on the mechanism of torilin action against inflammation is limited. This study aimed at determining the anti-inflammatory property of torilin in LPS-induced inflammation using in vitro model of inflammation. We examined torilin's effect on expression levels of inflammatory mediators and cytokines in LPS-stimulated RAW 264.7 macrophages. The involvement of NF-kB and AP-1, MAP kinases, and adaptor proteins were assessed. Torilin strongly inhibited LPS-induced NO release, iNOS, PGE2, COX-2, NF-α, IL-1ß, IL-6, and GM-CSF gene and protein expressions. In addition, MAPKs were also suppressed by torilin pretreatment. Involvement of ERK1/2, P38MAPK, and JNK1/2 was further confirmed by PD98059, SB203580, and SP600125 mediated suppression of iNOS and COX-2 proteins. Furthermore, torilin attenuated NF-kB and AP-1 translocation, DNA binding, and reporter gene transcription. Interestingly, torilin inhibited TAK1 kinase activation with the subsequent suppression of MAPK-mediated JNK, p38, ERK1/2, and AP-1 (ATF-2 and c-jun) activation and IKK-mediated I-κBα degradation, p65/p50 activation, and translocation. Together, the results revealed the suppression of NF-κB and AP-1 regulated inflammatory mediator and cytokine expressions, suggesting the test compound's potential as a candidate anti-inflammatory agent.

Effects of fluoroquinolone eye solutions without preservatives on human corneal epithelial cells in vitro.

AIMS: To evaluate the biologic effects of fluoroquinolone eye solutions without preservatives on cultured human corneal epithelial cells in vitro. METHODS: We studied the effect of diverse generations of topical fluoroquinolones such as ofloxacin 0.3%, levofloxacin 0.5%, tosufloxacin 0.3%, moxifloxacin 0.5% and gatifloxacin 0.3% on cultured human corneal epithelial cells. MTT-based calorimetric assay, lactate dehydrogenase leakage (LDH) assay and scratch wound test were performed. Corneal epithelial cell morphologies were examined by performing inverted light microscopy and transmission electron microscopy. RESULTS: In all topical fluoroquinolones, the metabolic activity of the corneal epithelial cells decreased in a time-dependent fashion and the LDH titer increased with longer exposure times. Especially, the LDH titers significantly increased after exposure to moxifloxacin 0.5% and gatifloxacin 0.3% compared with controls. The migration rates of the corneal epithelial cells were faster in ofloxacin 0.3% or levofloxacin 0.5% than other fluoroquinolones. Severe cellular morphological damage was observed after exposure to moxifloxacin 0.5% and gatifloxacin 0.3%. CONCLUSIONS: As moxifloxacin 0.5% and gatifloxacin 0.3% induced the toxic effect to the corneal epithelial cells, compared with other fluoroquinolones, the 4th fluoroquinolone eye solutions should be carefully used in case of the corneal epithelium is damaged by long duration of treatment or overdosage.

A randomised controlled trial comparing a thermal massager with artificial teardrops for the treatment of dry eye.

BACKGROUND: To evaluate the efficacy and safety of a thermal massager for the treatment of dry eye syndrome. METHODS: Ninety-five patients with dry eye syndrome were randomly assigned to receive either the thermal massager or artificial tears treatment. Thermal massage consisted of vibration, massage and thermotherapy and was carried out twice daily. Patients in the artificial tears group received 0.1% sodium hyaluronate solution five times daily. The Ocular Surface Disease Index (OSDI) score, break-up time (BUT), Schirmer test, fluorescein staining of the cornea, tear osmolarity test and adverse events were evaluated after 4 weeks. RESULTS: OSDI showed a significant improvement in both groups and improvement was significantly greater in thermal massager group (p=0.032). BUT and fluorescein staining also indicated significant improvement. No differences were found between the two groups in measures other than the OSDI. Adverse events were mild and transient. CONCLUSIONS: Thermal massage was effective in improving dry eye syndrome both subjectively and objectively. It was safe and seems to be a useful treatment option.

Torilin ameliorates type II collagen-induced arthritis in mouse model of rheumatoid arthritis.

Advancements in rheumatoid-arthritis-(RA) therapies have shown considerable progresses in the comprehension of disease. However, the development of new potential agents with relative safety and efficacy continues and natural compounds have been considered as alternatives to identify new entities. Since previous in-vivo data and our in-vitro findings showed that torilin has a strong anti-inflammatory property, we further investigated its effect against collagen-induced-arthritis-(CIA) in mice. CIA-induced DBA/1J mice were treated with torilin or methotrexate (MTX) for 5-weeks. Arthritis severity was evaluated by arthritic score and joint histopathology. Draining lymph node (dLN), joint and peripheral-blood mononuclear-cell (PBMC) counts, and activation/localization of T-/B-lymphocytes, dendritic cells (DCs) and neutrophils were examined by FACS analysis. Serum anti-type-II-collagen-(CII) antibody levels and cultured-splenocyte and serum cytokines were also evaluated. Torilin markedly reduced CIA-induced arthritic score, histopathology and leukocyte counts. Besides, torilin suppressed CIA-activated T-cells including CD3+, CD3+/CD69+, CD8+, CD4+ and CD4+/CD25+ in dLNs or joints. It also modified CD19+ or CD20+/CD23+ (B-cells), MHCII+/CD11c+ (DCs) and Gr-1+/CD11b+ (neutrophil) subpopulations. It further depressed total anti-CII-IgG, anti-CII-IgG1 and anti-CII-IgG2a antibody productions. Moreover, while IFN-γ and IL-10 were not affected, torilin suppressed CIA-induced serum TNF-α, IL-1ß and IL-6 levels. Interestingly, torilin also blocked IFN-γ, IL-17 and IL-6 cytokines while it did not affect IL-10 but enhanced IL-4 in splenocytes. These results show that torilin attenuated arthritis severity, modified leukocyte activations in dLNs or joints, and restored serum and splenocyte cytokine imbalances. Torilin may have immunomodulatory and anti-inflammatory properties with the capacity to ameliorate the inflammatory response in CIA-mice.

Photoreceptor layer assessed in tissue layer image using spectral-domain optical coherence tomography after surgical closure of macular hole.

PURPOSE: To investigate changes in the photoreceptor layer in tissue layer images using spectral-domain optical coherence tomography after macular hole surgery. METHODS: Visual acuity and spectral-domain optical coherence tomography scans were obtained in 24 eyes with surgically closed macular holes prospectively at 1, 2, 4, and 6 months after surgery. The scanned data were processed to generate tissue layer images of the photoreceptor layer, namely, photoreceptor layer map. Hyporeflective area and reflectivity at the fovea were analyzed to find a correlation with visual acuity. RESULTS: Mean visual acuity improved from 48.1 letters preoperatively to 62.5 letters at 1 month and to 73.8 letters at 6 months. Hyporeflective area in the photoreceptor layer map corresponded to attenuated signals from the junction of the inner and outer segments of the photoreceptors. Hyporeflective area decreased from 1.54 mm at 1 month to 0.60 mm at 6 months. Foveal reflectivity increased from 0.77 to 0.85. Visual acuity correlated with the hyporeflective area (r = 0.43-0.63) and foveal reflectivity (r = 0.35-0.61). CONCLUSION: After macular hole closure, attenuated signal from the junction of the inner and outer segments of the photoreceptors representing disorganization of the photoreceptor layer was shown in the photoreceptor layer map as a hyporeflective area, which correlated with the postoperative visual acuity. Hyporeflectivity decreased gradually with concurrent improvement in visual acuity.

The effect of epiblepharon surgery on visual acuity and with-the-rule astigmatism in children.

PURPOSE: To evaluate the effect of epiblepharon surgery on visual acuity and with-the-rule astigmatism in children compared to patients without surgical treatment. METHODS: We undertook a retrospective case control study and reviewed the charts of 202 eyes treated with epiblepharon surgery and of 142 eyes without surgery. The surgical procedure for epiblepharon correction used rotating suture techniques. Data regarding age, best corrected visual acuity, and degree of astigmatism were recorded. Baseline and 1-, 3-, 6-, and 12-month postoperative data were collected. The chi-square test, Student's t-test and general linear model analysis for repeated measures were applied. RESULTS: The mean astigmatism in the surgical group decreased from 1.10 ± 1.02 diopter (D) preoperatively to 0.84 ± 1.05 D at 3 months after surgery (p < 0.05). However, there was no statistically significant difference compared to the non-surgical group during the first year. The general linear model analysis comparing the mean astigmatism between the two groups over time showed a significant group-time interaction (p < 0.05). Within the surgical group, the higher baseline astigmatic subgroup and the 5- to 8-year-old group demonstrated greater cylinder reduction over time. The change in mean visual acuity was not significant in either group. CONCLUSIONS: Significant astigmatic reduction was found after surgical correction in epiblepharon patients. Patients with higher baseline astigmatism exhibited greater astigmatic reduction after epiblepharon surgery. These results suggest that, in order to reduce astigmatism, an epiblepharon operation should be considered in patients with a high level of astigmatism.

Production of Ginsenoside-Rg3 from Lipomyces starkeyi Grown on Ginseng-Steaming Effluent.

To produce ginsenoside-Rg3 enriched yeast from ginseng-steaming effluent (GSE), Lipomyces starkeyi, which tends to grow well in GSE, was cultured in sterilized GSE and its growth and production of ginsenoside-Rg3 were determined. Growth of L. starkeyi was 86.1 mg per g GSE and its ginsenoside-Rg3 contents was 0.013 mg per g GSE.

Stimulatory effect of dietary red ginseng on epidermal hydration and ceramide levels in ultraviolet-irradiated hairless mice.

Ultraviolet (UV) irradiation induces skin dryness, largely by disruption of the epidermal barrier. In a search for dietary and plant compounds that would protect against skin dryness, we investigated the dietary effect of red ginseng (the steamed root of Panax ginseng C.A. Meyer) on epidermal levels of hydration and ceramides, the most important lipids for maintaining the epidermal barrier, in UV-irradiated mice. Albino hairless mice were fed either control diets (group UV [UV-irradiated control]) or diets with 0.5% (group H0.5) or 1% (group H1.0) red ginseng extract for 5 weeks in parallel with UV irradiation. A normal control group (group C) was fed a control diet without UV irradiation for 5 weeks. Skin dryness in group UV, as assessed by epidermal levels of hydration and ceramides, was significantly lower than those in group C. With no differences in food intake and weight gains among groups, epidermal levels of hydration and ceramides in group H0.5 were similar to those in group C. In addition, protein expression of serine palmitoyltransferase (SPT), a key enzyme involved in de novo ceramide synthesis, was increased in group H0.5. However, epidermal levels of hydration and ceramides in group H1.0 did not differ from those in group UV, in which ceramidase, an enzyme involved in ceramide degradation, was highly expressed. In conclusion, we demonstrate that dietary supplementation of 0.5% red ginseng protects skin from UV-induced dryness with an accumulation of ceramides due to elevated expression of SPT protein.

Red ginseng root extract mixed with Torilus fructus and Corni fructus improves facial wrinkles and increases type I procollagen synthesis in human skin: a randomized, double-blind, placebo-controlled study.

Red ginseng contains many bioactive constituents, including various ginsenosides that are believed to have antioxidant, immunostimulatory, and anti-aging activities. Yet, no controlled human study has explored its effects on photoaged skin. This study determined whether long-term intake of a red ginseng extract-containing Torilus fructus and Corni fructus mixture reduces facial wrinkles and increases collagen synthesis in human skin. Healthy female volunteers over 40 years of age were randomized in a double-blind fashion to receive either red ginseng extract-containing herbal mixture at 3 g/day or placebo for 24 weeks. Facial wrinkles, elasticity, epidermal water content, erythema, and pigmentation were measured objectively. Facial skin samples were taken before and after treatment, and real-time polymerase chain reaction and immunohistochemical analyses were undertaken for expression of type I procollagen, matrix metalloproteinase (MMP)-9, and fibrillin-1, which are wrinkle-related biochemical markers. A total of 82 subjects completed the study. Facial wrinkles were significantly improved, type I procollagen gene and protein expression was increased, MMP-9 gene induction was prevented, and fibrillin-1 fiber length was elongated only in the treatment group. No changes were seen in the facial elasticity, epidermal water content, facial erythema and pigmentation, and epidermal thickness in either group. Thus a red ginseng extract-containing Torilus fructus and Corni fructus mixture improves facial wrinkles, a clinical sign of photoaging, and this improvement is associated with biochemical and histological evidence of increased collagen synthesis in the dermis. These results substantiate the alleged beneficial effects of red ginseng on photoaging and support its use as an effective "beauty food."

Effect of ginseng mixture on osteoporosis in ovariectomized rats.

In this study, the authors have characterized the effect of HER-S (red ginseng, Angelicae gigantis Radix, Phyllostachys folium, and soybean extracts) on osteoporosis-associated phenomena in ovariectomized (OVX) rats by measuring body weights and bone histomorphometries in control, sham, OVX, OVX(beta-estradiol-treated), and OVX(HER-S-treated) rats. Light microscopic analyses showed a porous or eroded appearance on the femoral trabecular bone surface in OVX rats, whereas the femoral trabecular bone surfaces of the other groups (control, sham, OVX(17beta-estradiol-treated), and OVX(HER-S-treated) rats) were composed of fine particles. The femoral trabecular bone area and number were decreased in OVX rats, but these reductions were significantly prevented by the administration of HER-S for 7 weeks, similar to estrogen. In the blood biochemistry results, serum phosphorus, calcium, T(3), and T(4) remained unchanged, but blood estrogen levels were significantly increased in HER-S-treated rats, which suggests that estrogen is related to the mechanism of the HER-S-induced antiosteoporosis function in OVX rats.

Physiological Functionality and Enzyme Activity of Biomass from Pichia anomala Grown on Ginseng-Steaming Effluent.

A novel biomass was prepared from Pichia anomala KCCM 11473, which grew well in ginseng-steaming effluent (GSE), and its physiological functionalities and enzyme activities were determined. When the strain was cultured in the GSE (pH 6.0) at 30℃ for 48 h, 1.6 mg of biomass per ml-cultures was produced. The cell-free extract of the biomass showed high antihypertensive angiotensin I-converting enzyme inhibitory activity of 72.0% and anticholesteromia HMG-CoA reductase inhibitory activity of 46.5%. The cell-free extract also showed 13.0 U per ml and 8.5 U per ml of neutral protease activity and alkaline protease, respectively.

Production of Antihypertensive Angiotensin I-Converting Enzyme Inhibitor from Malassezia pachydermatis G-14.

To produce a novel antihypertensive angiotensin I-converting enzyme (ACE) inhibitor from yeast, a yeast isolate, designated G-14 showing the highest ACE inhibitory activity was obtained and identified as Malassezia pachydermatis based on morphological, biochemical and cultural characteristics. The maximal extracellular ACE inhibitor production was obtained from M. pachydermatis G-14 when the strain was cultured in YEPD medium containing 0.5% yeast extract, 3.0% peptone and 2.0% glucose at 30℃ for 24 h and the final ACE inhibitory activity was 48.9% under the above condition.