Etch characteristics of maskless oxide/nitride/oxide/nitride (ONON) stacked structure using C4H2F6-based gas.

Oxide/Nitride/Oxide/Nitride (ONON; SiO2/SiNx/SiO2/SiNx) stacked structure is widely used in the 3D vertical structure of semiconductor cells. Previously, to form a 3D cells, photoresist (PR) was patterned and repeatedly trimmed on the top of ONON after the etching of one ON layer. Due to the time-consuming process of etching layer-by-layer of ON layer, two-step etch processing using C4F8-based or C4F6-based gases composed of maskless ONON stack feature etching and followed one ON layer-by layer etching by PR trimming in the ONON stack feature are employed these days. However, the two-step etching method resulted in poor etch profiles of maskless ONON stack feature in addition to high global warming potential of C4F8 and C4F6. In this study, we investigated the etching of maskless ONON stack feature using C4H2F6-based gas having a low global warming potential and the effects of C4H2F6-based gas on the etch characteristics of maskless ONON stack feature such as etch rate, etch profile, change in critical dimensional (CD), and etch selectivity between SiO2 and SiNx have been investigated. C4H2F6-based gas showed the highest etch rates compared to C4F6 and C4F8-based gases in addition to the etch selectivity of ~ 1:1 between SiO2 and SiNx due to hydrogen included in the gas structure. In addition, the change in horizontal CD was lower in the order of C4H2F6, C4F6, and C4F8-based gases due to the more effective sidewall passivation in the order of C4F8, C4F6, and C4H2F6-based gases. The thicker carbon-based polymer layer on the sidewall also played an important role in maintaining the shape of the top edge shape of maskless ONON stack feature when etching a line feature in an environment without a mask.

Stability of clockwise rotation of the maxillary occlusal plane in skeletal Class III patients treated with two-jaw surgery.

OBJECTIVE: The objective of the study was to investigate the post-surgical stability of clockwise rotation of the maxillary occlusal plane (MXOP) after Le Fort I osteotomy for posterior impaction and advancement, in skeletal Class III patients who had undergone two-jaw orthognathic surgery (2J-OGS). MATERIALS AND METHODS: The sample set consisted of 46 patients (18 males and 28 females). Using lateral cephalograms taken at the initial evaluation (T0), before 2J-OGS (T1), after 2J-OGS (T2) and at debonding (T3), the amount of MXOP change (criterion: 2° in ΔMXOP [between T2 and T3]) was used to assign patients to Group 1 (G1, high relapse, -3.09°) or Group 2 (G2, low relapse, -0.99°). Findings were statistically compared between the groups. RESULTS: Compared with G2, G1 exhibited more severe skeletal Class III relationships and a flatter MXOP at T0, a greater increase in ΔANB, and more clockwise rotation of ΔMXOP and ΔFMA between T1 and T2. G1 also showed a greater decrease in ΔANB, higher counterclockwise rotation of ΔMXOP and upward movement of U1 between T2 and T3. Regression analysis yielded the following equation: MXOP (ΔT2-T3) = -0.37 X MXOP (ΔT1-T2) -0.43. CONCLUSIONS: The higher the clockwise rotation of the MXOP during 2J-OGS in skeletal Class III patients, the greater the subsequent relapse (counterclockwise rotation) of the MXOP.

Development of Squalene-Based Oil-in-Water Emulsion Adjuvants Using a Self-Emulsifying Drug Delivery System for Enhanced Antigen-Specific Antibody Titers.

Introduction: A recombinant protein cannot induce sufficient immune response by itself. Various substances, including cytokine and mineral, have been used as adjuvants to enhance the immunogenicity and efficacy of vaccines; however, most of them induce excessive immune responses or exhibit cytotoxicity. In this study, a self-emulsifying drug delivery system (SEDDS), an isotropic mixture of oil, surfactant, and solvent, was designed for oil-in-water emulsions as a non-toxic adjuvant to increase immune response to antigens. Methods: Squalene-based oil-in-water emulsions were prepared by SEDDS to assess its value as an adjuvant. Fifteen emulsions (F1-F15) were prepared by stirring two types of surfactants (Span® 85 and Kolliphor® RH40), and squalene and carboxymethyl cellulose (CMC) were added at different ratios. The physical properties and viscosity of the 15 emulsions were evaluated by measuring droplet size, zeta potential, and polydispersity index. The toxic effect of emulsions was assessed by acute toxicity test in mice. Mice were immunized twice with 1:1 mixtures of antigen and adjuvant (15 emulsions, phosphate-buffered saline, and commercial alum-based adjuvant). Antigen-specific antibody titers from immunized mice serum were measured by an indirect enzyme-linked immunosorbent assay. Results: All emulsions exhibited droplet sizes ranging from 322 to 812 nm and maintained zeta potential values between -30 mV to -10 mV for 4 weeks, indicating good physical stability as a vaccine adjuvant. Additionally, all emulsions were non-toxic, and they induced humoral immunity at a similar level compared to commercial alum-based adjuvant in the first immunization. However, 12% squalene-based oil-in-water emulsion containing 0.5% of ultra-high viscosity CMC (F15) showed significantly higher immune response than a commercial adjuvant in the second immunization. Conclusion: Squalene-based oil-in-water emulsions could be conveniently prepared using SEDDS technique and are non-toxic and stable at room temperature storage. Moreover, squalene-based oil-in-water emulsions show enhanced immune induction with antigen; hence, they can possibly be used as effective adjuvants.

Inpatients with shoulder osteoarthritis who received integrative Korean medicine treatment: Long-term follow-up of an observation study.

This study aimed to investigate the long-term clinical efficacy of and satisfaction with integrative Korean medicine (KM) treatment in patients with shoulder osteoarthritis (SOA). We conducted a prospective observational study of patients with SOA. Patients aged 19 years and older who underwent inpatient treatment for more than 1 week were eligible for enrollment in the study. The primary evaluation index was the numeric rating scale for shoulder pain. Sub-evaluation indices included the Shoulder Pain and Disability Index for shoulder function, EuroQol-5-dimension score for overall quality of life, and Patient Global Impression of Change. Outcome measures were assessed at admission, discharge, and follow-up. For the follow-up questionnaire survey, the following information was collected: current status, surgery after discharge, reasons for finding integrative KM treatment satisfactory/unsatisfactory, and quality of life after discharge. In total, 186 patients were enrolled in the primary analysis, and 103 patients completed the follow-up survey. The mean number of days of follow-up was 1019 ±â€…439. Compared with the baseline, the mean differences in the numeric rating scale and Shoulder Pain and Disability Index were 3.05 ±â€…0.34 and 36.06 ±â€…5.53, respectively. Regarding the Patient Global Impression of Change, 89 out of 103 (86.4%) patients chose "minimally improved" or better. Furthermore, the EuroQol-5-dimension score also increased, showing an improvement of health-related quality of life after treatment. Integrative KM treatment is a potential option for reducing pain severity and improving function and health-related quality of life in patients with SOA. Prospective randomized studies would support this finding for the next step.

Swelling-based preparation of polypropylene nanocomposite with non-functionalized cellulose nanofibrils.

Dispersion of nanofillers in a polymer matrix is one of the most important steps in preparing polymer nanocomposites. However, hydrophobic polymers and hydrophilic nanofillers are intrinsically incompatible, making it difficult to mix them homogeneously. Here, we propose the swelling-based particle adsorption method (SPA) providing a simple route to disperse cellulose nanofibrils (CNFs) within incompatible polypropylene (PP) matrix without surface functionalization of CNFs. The SPA enables CNFs to adsorb onto the surface of PP particles using a small amount of solvent. PP/CNFs composite films fabricated from the SPA showed increased Young's modulus by 80%, which agrees well with a theoretical prediction proving nano-dispersed. Furthermore, simply mixing a bit of polypropylene-graft-maleic anhydride can improve the tensile strength by 30% and the elongation at break by 10-fold than that of PP/CNFs composites. The SPA can be universally applied to any incompatible polymer-nanofiller pairs for the fabrication of nanocomposite materials.

A case report of a patient presented with skin ulcer after treatment of lenvatinib.

A 60-year-old man diagnosed with unresectable hepatocellular carcinoma (HCC) presented to the hospital with pain in the perineal region. He had been taking lenvatinib every day for 2 months after he was diagnosed with HCC with metastases to the lymph node, small bowel mesentery, and retroperitoneal space. Enhanced abdominal computed tomography revealed mild elevation in intensity in the perineal subcutaneous tissue with subcutaneous emphysema. The patient was diagnosed with Common Terminology Criteria for Adverse Events grade 3, skin ulceration of stage IV with full-thickness skin loss and tissue necrosis in the muscular layer. The patient was taken off the medication with prescription of antibiotics, and after 3 weeks, the skin has fully recovered. This is the first report of an HCC patient who presented with a skin ulceration of stage IV after lenvatinib treatment. We recommend stopping the medication immediately and changing to alternative treatments with appropriate supportive care.

Dietary mercury intake and colorectal cancer risk: A case-control study.

BACKGROUND & AIMS: The main source of mercury exposure is food such as fish and shellfish. Mercury is a growing concern due to its associations with a number of harmful health effects, including cancer. The objectives of this study were to examine the association between dietary mercury intake and colorectal cancer (CRC) risk and to determine whether this association differs by anatomical site and menopausal status. METHODS: A case-control study was conducted with 2769 participants (923 cases and 1846 controls) in Korea. Dietary mercury intake and fish and shellfish consumption were assessed using a semiquantitative food frequency questionnaire. RESULTS: A high intake of dietary mercury was associated with an increased risk of CRC (in the group with lower fish and shellfish consumption; odds ratio (OR): 3.13; 95% confidence interval (95% CI): 2.33, 4.71, in the group with higher fish and shellfish intake; OR: 3.84, 95% CI: 2.20, 7.30) after adjusting for all potential confounders by anatomic site in men. Among women, the results differed by fish and shellfish consumption and menopausal status. Regarding the amount of fish and shellfish intake, a positive association was found only in the group with lower intake (CRC; OR: 2.24, 95% CI: 1.36, 3.72, colon cancer; OR: 2.25, 95% CI: 1.22, 4.16, rectal cancer; OR: 2.28, 95% CI: 1.13, 4.57). In the stratified analysis by menopausal status, the elevated risk of CRC was still observed among both pre- and postmenopausal women depending on anatomical site, except for the colon cancer patients with premenopausal status. CONCLUSIONS: A high intake of mercury was associated with an elevated risk of overall CRC. Future large-scale prospective cohort studies are recommended to investigate the causal effects of dietary mercury intake by fish and shellfish consumption on CRC risk depending on anatomical site and menopausal status.

Prognostic role of the beta-2 adrenergic receptor in clear cell renal cell carcinoma.

The beta-2 adrenergic receptor (ADRB2) regulates the proliferation, apoptosis, angiogenesis, migration, and metastasis of cancer cells. However, its function in the progression of clear cell renal cell carcinoma (ccRCC) is unknown. Here, we report that ADRB2 can be a novel prognostic factor for patients with ccRCC. The differential expression of ADRB2 in low-stage (stages I and II), high-stage (stages III and IV), low-grade (grades I and II), and high-grade (grades III and IV) ccRCC was identified in cohorts of patients from The Cancer Genome Atlas and the International Cancer Genome Consortium. We evaluated ADRB2 expression as a prognostic factor using the Kaplan-Meier survival curve, multivariate analysis, time-dependent area under the curve (AUC) of Uno's C-index, and AUC of the receiver operating characteristics (ROC) at five years. Kaplan-Meier analysis revealed that reduced ADRB2 expression is associated with poor prognosis in ccRCC patients. Analysis of C-indices and AUC-ROC further confirmed this result. Moreover, multivariate analysis confirmed the prognostic significance of ADRB2 expression. Collectively, these findings suggest that ADRB2 is a potential prognostic factor for ccRCC.

Association between dietary cadmium intake and early gastric cancer risk in a Korean population: a case-control study.

PURPOSE: Foods such as grains and vegetables are the dominant sources of exposure to cadmium, which has been classified as a carcinogen by various public health agencies. Cadmium exposure is a growing concern due to its associations with numerous harmful health effects, including gastric cancer risk. The objective of this study was to investigate the association of dietary cadmium intake and the consumption of cadmium-contributing foods with early gastric cancer risk. METHODS: A case-control study including 1245 subjects (cases, 415; controls, 830) was conducted in Korea. The dietary cadmium intake and the consumption of cadmium-contributing foods were assessed using a semi-quantitative food frequency questionnaire. RESULTS: After adjustment for covariates, the gastric cancer risk was increased for participants in the highest tertile of cadmium intake [odds ratios (ORs) 1.33, 95% confidence intervals (95% CIs) 0.94-1.88], but there was no significance. Both female (ORs 2.71, 95% CIs 1.37-5.36) and male (ORs 1.63, 95% CIs 1.07-2.50) participants in the highest tertile of rice consumption had a higher gastric cancer risk than did those in the lowest tertile. Men in the highest tertile of crab consumption had a gastric cancer risk 2.23 times greater than that of men in the lowest tertile (ORs 2.23, 95% CIs 1.21-4.13), but a difference was not seen in women. CONCLUSIONS: Future studies examining the causal effects of dietary cadmium intake and the consumption of cadmium-contributing foods on early gastric cancer risk in large-scale prospective cohorts are recommended.

Comparison of the effects of desflurane and total intravenous anesthesia on the optic nerve sheath diameter in robot assisted laparoscopic radical prostatectomy: A randomized controlled trial.

BACKGROUND: Optic nerve sheath diameter (ONSD) is a well-known surrogate marker for intracranial pressure during robot-assisted laparoscopic radical prostatectomies (RALP). ONSD during RALP is known to increase due to elevated intracranial pressure as a result of the steep Trendelenburg position and carbon dioxide pneumoperitoneum. We aimed to compare the effects of total intravenous anesthesia (TIVA) and desflurane anesthesia (DES) on ONSD during RALP. METHODS: Patients scheduled for RALP were enrolled and randomly assigned to the TIVA (propofol and remifentanil) or DES (desflurane and remifentanil) group in this randomized trial. Ultrasonographic measurements of ONSD were conducted before administration of anesthesia (T0), 10 minutes after the Trendelenburg position (T1), 1 hour after the Trendelenburg position (T2), 2 hours after the Trendelenburg position (T3), 10 minutes after resuming the supine position (T4), and at the time of arrival in the post-anaesthetic care unit (T5). The primary outcome measure was the mean ONSD at T2 of the TIVA and DES group during RALP. RESULTS: A total of 56 patients were analysed in this study. The mean ONSD at T1, T2, T3, and T4 were significantly lower for patients in the TIVA group compared with those in the DES group (P = .023, .000, .000, and .003, respectively). CONCLUSION: The mean ONSD for patients in the TIVA group was significantly lower than that in the DES group during the RALP procedure. Our findings suggest that TIVA may be a more suitable anesthetic option for patients at risk of cerebral hypoperfusion.

Effect of dexamethasone on the onset time and recovery profiles of cisatracurium.

BACKGROUND: The effect of dexamethasone injection on cisatracurium-induced neuromuscular block was compared according to different injection time points. METHODS: One hundred seventeen patients were randomly assigned to three groups: 8 mg of dexamethasone injected intravenously 2-3 h before anesthesia (group A), just before anesthesia induction (group B), and at the end of surgery (control group). Three minutes after anesthesia induction, intubation was performed without neuromuscular blockers, and acceleromyography was initiated. All patients received 0.05 mg/kg cisatracurium; the onset time and recovery profiles were recorded. RESULTS: Eighty patients were finally enrolled. The onset time (median [interquartile range], seconds) was significantly hastened in group A (520.0 [500.0-560.0], n = 30) compared to that in group B (562.5 [514.0-589.0], n = 22) (P = 0.008) and control group (586.5 [575.0-642.5], n = 28) (P < 0.001). The onset time in group B was faster than the control group (P = 0.015). The recovery time [mean (95% CI) minutes] was significantly hastened in group A [28.5 (27.3-29.6)] compared to that in group B [32.3 (31.0-33.6)] (P < 0.001) and control group [30.9 (29.9-31.8)] (P = 0.015). The total recovery time was significantly hastened more in group A [47.1 (45.5-48.6)] than group B [52.8 (51.6-54.0) minutes] (P < 0.001) and control group [50.5 (48.7-52.3) minutes] (P = 0.008). CONCLUSIONS: A single dose of 8 mg of dexamethasone hastened the onset and total recovery times of cisatracurium-induced block by approximately 15 and 9%, respectively if administered 2-3 h prior to surgery.

Erratum: Correction of Funding Source: The Burden of Cancer in Korea during 2012: Finding from a Prevalence-Based Approach.

This corrects the article on p. S168 in vol. 31, PMID: 27775254.

Effects of Dendropanax morbifera Léveille extracts on cadmium and mercury secretion as well as oxidative capacity: A randomized, double-blind, placebo-controlled trial.

In this randomized, double-blind, placebo controlled clinical trial, the effects of Dendropanax morbifera (D. morbifera) Léveille on heavy metal (cadmium and mercury) excretion as well as on lipid peroxidation and Cu, Zn-superoxide dismutase (SOD1) activity were investigated. For this study, tablets containing placebo or 300 mg of the leaf extract from D. morbifera Léveille were used. A total of 60 eligible healthy subjects were enrolled in this randomized, double-blind, placebo-controlled study. The differences in cadmium, mercury, and malondialdehyde (MDA) levels and SOD1 activity were measured in the serum 60 days after treatment with placebo or D. morbifera Léveille extracts. No significant differences between baseline characteristics and biochemical values were identified in subjects in the placebo and D. morbifera Léveille groups. Serum levels of cadmium, mercury and MDA decreased following consumption of D. morbifera Léveille extracts; however, no significant differences were identified. In addition, female, but not male, subjects who consumed D. morbifera Léveille extracts showed a significant increase in SOD1 activity. This result suggests that chronic consumption of D. morbifera Léveille extract can help to facilitate excretion of cadmium and mercury from serum and increase the antioxidant capacity in humans.

Neuroprotection against 6-OHDA-induced oxidative stress and apoptosis in SH-SY5Y cells by 5,7-Dihydroxychromone: Activation of the Nrf2/ARE pathway.

AIMS: The aim of this study was to prove the neuroprotective effect of 5,7-Dihydroxychromone (DHC) through the Nrf2/ARE signaling pathway. To elucidate the mechanism, we investigated whether 6-hydroxydopamine (6-OHDA)-induced neurotoxicity in SH-SY5Y cells could be attenuated by DHC via activating the Nrf2/ARE signal and whether DHC could down-regulate 6-OHDA-induced excessive ROS generation MAIN METHODS: To evaluate the neuroprotective effect of DHC against 6-OHDA-induced apoptosis, FACS analysis was performed using PI staining. The inhibitory effect of DHC against 6-OHDA-induced ROS generation was evaluated by DCFH-DA staining assay. Additionally, translocation of Nrf2 to the nucleus and increased Nrf2/ARE binding activity, which subsequently resulted in the up-regulation of the Nrf2-dependent antioxidant gene expressions including HO-1, NQO1, and GCLc, were evaluated by Western blotting and EMSA. KEY FINDINGS: Pre-treatment of DHC, one of the constituents of Cudrania tricuspidata, significantly protects 6-OHDA-induced neuronal cell death and ROS generation. Also, DHC inhibited the expression of activated caspase-3 and caspase-9 and cleaved PARP in 6-OHDA-induced SH-SY5Y cells. DHC induced the translocation of Nrf2 to the nucleus and increased Nrf2/ARE binding activity which results in the up-regulation of the expression of Nrf2-dependent antioxidant genes, including HO-1, NQO1, and GCLc. The addition of Nrf2 siRNA abolished the neuroprotective effect of DHC against 6-OHDA-induced neurotoxicity and the expression of Nrf2-mediated antioxidant genes. SIGNIFICANCE: Activation of Nrf2/ARE signal by DHC exerted neuroprotective effects against 6-OHDA-induced oxidative stress and apoptosis. This finding will give an insight that activating Nrf2/ARE signal could be a new potential therapeutic strategy for neurodegenerative disease.

Isoflavones with neuroprotective activities from fruits of Cudrania tricuspidata.

Ten isoflavones, cudraisoflavones B-K (1-10), together with 27 known isoflavones, were isolated from the EtOAc soluble extract of fruits of Cudrania tricuspidata. The structures of compounds 1-10 were elucidated on the basis of MS and NMR spectroscopic data, including 2D NMR experiments. Compounds 7-9 and three known (11-13) compounds showed neuroprotective activity against 6-hydroxydopamine induced cell death in human neuroblastoma SH-SY5Y cells, with EC50 values of 0.5-9.2 µM.

Dendropanax morbifera Léveille extract facilitates cadmium excretion and prevents oxidative damage in the hippocampus by increasing antioxidant levels in cadmium-exposed rats.

BACKGROUND: Dendropanax morbifera Léveille is used in herbal medicine as a cancer treatment. In this study, we investigated the effects of Dendropanax morbifera stem extract (DMS) on cadmium (Cd) excretion from the blood and kidney and brain tissues of rats exposed to cadmium, as well as the effects of DMS on oxidative stress and antioxidant levels in the hippocampus after Cd exposure. METHODS: Seven-week-old Sprague-Dawley rats were exposed to 2 mg/kg of cadmium by intragastric gavage and were orally administered 100 mg/kg of DMS for 4 weeks. Animals were sacrificed and Cd determination was performed using inductively coupled plasma mass spectrometry. In addition, the effects of Cd and/or DMS on oxidative stress were assayed by measuring reactive oxygen species production, protein carbonyl modification, lipid peroxidation levels, and antioxidant levels in hippocampal homogenates. RESULTS: Exposure to Cd significantly increased Cd content in the blood, kidneys, and hippocampi. DMS treatment significantly reduced Cd content in the blood and kidneys, but not in the hippocampi. Exposure to Cd significantly increased reactive oxygen species production, protein carbonyl modification, lipid peroxidation, total sulfhydryl content, reduced glutathione content, and glutathione reductase activity. In contrast, Cu, Zn-superoxide dismutase (SOD1), catalase (CAT), glutathione peroxidase (GPx), and glutathione-S-transferase (GST) activity in the hippocampus were significantly decreased after exposure to Cd, and administration of DMS significantly inhibited these Cd-induced changes. CONCLUSION: These results indicate that DMS facilitates cadmium excretion from the kidneys, reduces cadmium-induced oxidative stress in the hippocampus, and modulates SOD1, CAT, GPx, and glutathione-S-transferase activities.

Neuroprotective Xanthones from the Root Bark of Cudrania tricuspidata.

Seventeen new prenylated xanthones (1-17) were isolated from an ethyl acetate-soluble extract of root bark of Cudrania tricuspidata together with 17 previously identified xanthones. The structures of the new compounds were elucidated by spectroscopic methods. Six new compounds (3, 7, 8, 9, 15, and 16) and six known compounds (18-23) showed neuroprotective effects against 6-hydroxydopamine-induced cell death in human neuroblastoma SH-SY5Y cells, with EC50 values of 0.7-16.6 µM.

Onion peel water extracts enhance immune status in forced swimming rat model.

Onion peel contains a high concentration of quercetin and other flavonoids. In this study, the potential immune-enhancing effects of an onion peel water extract (OPE) supplement were investigated by the rat forced swimming test. OPE was prepared using hot water. Thirty-six male Sprague Dawley rats were fed a pellet diet for 1 week and were then randomly divided into six groups: normal control (NC), forced swimming control (FSC), positive control (quercetin 20 mg/kg), and three groups administered 4, 20, or 100 mg/kg of OPE. Oral drug administration was conducted daily for 4 weeks. All rats, except those of NC group, were forced to swim in water and were considered exhausted when they failed to rise to the water surface to breathe within a 7-s period. Blood lymphocyte counts, immune organ weights, histopathological analysis, and serum interferon (IFN)-γ, tumor necrosis factor (TNF)-α, and interleukin (IL)-12 levels were determined. OPE-treated rats consumed more food and had an increased thymic cortex to medulla ratio than that observed in FSC group rats (P<0.05). The area of the white pulp in the spleens of OPE-treated group rats was increased compared with that in FSC group rats (P<0.05). Furthermore, blood lymphocyte numbers and IFN-γ, TNF-α, and IL-12 concentrations were significantly higher in OPE-fed groups than in FSC group (P<0.05). These results suggest that an OPE supplement can improve the immune status by increasing the number of immune-related cells and specific cytokine levels.

Reynosin protects against neuronal toxicity in dopamine-induced SH-SY5Y cells and 6-hydroxydopamine-lesioned rats as models of Parkinson's disease: Reciprocal up-regulation of E6-AP and down-regulation of α-synuclein.

Aggregation of α-synuclein (ASYN) is considered a major determinant of neuronal loss in Parkinson's disease (PD). E6-associated protein (E6-AP), an E3 ubiquitin protein ligase, has been known to promote the degradation of α-synuclein. The aim of this study was to assess the effects of the sesquiterpene lactone reynosin on dopamine (DA)-induced neuronal toxicity and regulation of E6-associated protein and α-synuclein proteins in both in vitro and in vivo models of Parkinson's disease. Usi"ng flow cytometry and western blot analysis, we determined that reynosin significantly protected both against cell death from dopamine-induced toxicity in human neuroblastoma SH-SY5Y cells and against the loss of tyrosine hydroxylase (TH)-positive cells in 6-hydroxydopamine (6-OHDA)-lesioned rats (a rodent Parkinson's disease model system). In addition, reynosin made up-regulation of E6-associated protein expression and down-regulation of the over-expression of α-synuclein protein in both dopamine-treated SH-SY5Y cells and 6-hydroxydopamine-lesioned rats. These results suggest that the protective effect of reynosin against dopamine-induced neuronal cell death may be due to the reciprocal up-regulation of E6-associated protein and down-regulation of α-synuclein protein expression.

Dual inhibition of γ-oryzanol on cellular melanogenesis: inhibition of tyrosinase activity and reduction of melanogenic gene expression by a protein kinase A-dependent mechanism.

The in vitro effects on melanogenesis of γ-oryzanol (1), a rice bran-derived phytosterol, were investigated. The melanin content in B16F1 cells was significantly and dose-dependently reduced (-13% and -28% at 3 and 30 µM, respectively). Tyrosinase enzyme activity was inhibited by 1 both in a cell-free assay and when analyzed based on the measurement of cellular tyrosinase activity. Transcriptome analysis was performed to investigate the biological pathways altered by 1, and it was found that gene expression involving protein kinase A (PKA) signaling was markedly altered. Subsequent analyses revealed that 1 stimulation in B16 cells reduced cytosolic cAMP concentrations, PKA activity (-13% for cAMP levels and -40% for PKA activity), and phosphorylation of the cAMP-response element binding protein (-57%), which, in turn, downregulated the expression of microphthalmia-associated transcription factor (MITF; -59% for mRNA and -64% for protein), a key melanogenic gene transcription factor. Accordingly, tyrosinase-related protein 1 (TRP-1; -69% for mRNA and -82% for protein) and dopachrome tautomerase (-51% for mRNA and -92% for protein) in 1-stimulated B16F1 cells were also downregulated. These results suggest that 1 has dual inhibitory activities for cellular melanogenesis by inhibiting tyrosinase enzyme activity and reducing MITF and target genes in the PKA-dependent pathway.