Poly-dipeptides produced from C9orf72 hexanucleotide repeats cause selective motor neuron hyperexcitability in ALS.

SignificanceThe GGGGCC hexanucleotide repeat expansion in the chromosome 9 open reading frame 72 (C9orf72) gene is the most common genetic cause of amyotrophic lateral sclerosis (ALS). Despite myriad studies on the toxic effects of poly-dipeptides produced from the C9orf72 repeats, the mechanisms underlying the selective hyperexcitability of motor cortex that characterizes the early stages of C9orf72 ALS patients remain elusive. Here, we show that the proline-arginine poly-dipeptides cause hyperexcitability in cortical motor neurons by increasing persistent sodium currents conducted by the Nav1.2/ß4 sodium channel complex, which is highly expressed in the motor cortex. These findings provide the basis for understanding how the C9orf72 mutation causes motor neuron hyperactivation that can lead to the motor neuron death in C9orf72 ALS.

Microbial assessment of medicinal herbs (Cnidii Rhizoma and Alismatis Rhizoma), effects of electron beam irradiation and detection characteristics.

Medicinal herbs comprise of heavy microbial contaminations. This study aimed to assess microbial hazards including foodborne pathogens in 20 commercial medicinal herbs, Cnidii Rhizoma (C1-C10) and Alismatis Rhizoma (T1-T10) as well as to evaluate irradiation effects of E-beam on microbial load and detection chracteristics. Four samples (C5, C10, T1, T8) from both herbs with higher microbial load were selected for evaluating the irradiation effect of E-beam (up to 10 kGy) on microbial load and radiation-induced changes in detection markers by standard methods (Codex, Korean Food Code), such as direct epifluorescent filter technique/aerobic plate count (DEFT/APC), photostimulated luminescence (PSL), thermoluminescence (TL), and electron spin resonance (ESR). DEFT/APC revealed non-evidence of pre-sterilization of all samples. PSL differentiated irradiated samples (1, 5, and 10 kGy) of both herbs from non-irradiated (control: 0 kGy). Both TL and ESR methods validated PSL screening results by detecting radiation-induced markers from E-beam irradiated medicinal herbs.

Influence of E-beam irradiation on microbiological and physicochemical properties and fatty acid profile of frozen duck meat.

This study investigated the effect of different doses (0, 3, and 7 kGy) of e-beam on the microbiological and physicochemical qualities and the profile of fatty acids of the frozen duck meat (FDM). Electron beam at the dose of 3 kGy showed more than 2 log and 1 log cycles of reduction in the total bacterial (TAB) and coliform counts (TCC), respectively. The results indicated an increase in the TBARS values (1.50 ± 0.02 mg MDA/kg), peroxide value (0.83 ± 0.04 meq peroxide/kg), and total volatile base nitrogen (1.31 ± 0.16 mg/100 ml), but no effect on the sensory parameters. Irradiation lowered the lightness (L*) (31.87 ± 0.98) and redness (a*) (11.04 ± 0.20) values but elevated the metmyoglobin content in FDM. In addition, irradiation had no effect on the benzopyrene content; however, a reduction was observed in the vitamin A (0.239 ± 0.015 µg/g) and vitamin E (1.847 ± 0.075 µg/g) contents of the FDM samples. There were no trans-fatty acids present in the treated (irradiated) as well as the untreated (nonirradiated) meat samples (FDM), whereas the fatty acid content decreased in irradiated samples, in contrast with the nonirradiated control. Electronic nose clearly discriminated between the nonirradiated and irradiated FDM based on principal component analysis. It is concluded that the e-beam successfully improved the microbial quality of FDM with slight changes in physicochemical properties, but without altering its sensory properties.

E-sensing, calibrated PSL, and improved ESR techniques discriminate irradiated fresh grapefruits and lemons.

Grapefruit and lemon were exposed to phytosanitary irradiation doses of 0, 0.4, and 1 kGy, and then electronic-sensing screening of irradiation status and identification of radiation-induced ESR markers were conducted during 20 days storage. Codex standard photostimulated luminescence measurement (PSL1) was not a reliable indicator of irradiation status. Electronic tongue and electronic nose, however, showed potential as screening tools for discriminating irradiated fruits from non-irradiated counterparts based on principal component analysis of taste attributes and flavor patterns. Calibrated PSL approach clearly distinguished irradiated from non-irradiated samples based on the PSL ratio (PSL2/PSL1). Verification of irradiation status by electron spin resonance spectroscopy revealed clear paramagnetic centers from both irradiated fruits samples and ethanol-vacuum drying pretreatment improved radiation-induced ESR signal detection; not prominent enough in 0.4 kGy-irradiated commodities during 20 days of storage. Peel parts of both fruits showed high limonene which proportionally increased with irradiation doses (p < 0.05 kGy).

Effect of E-Beam Irradiation on Microbial Load, Stability of Active Components, and Anti-Inflammatory Activity of Cnidii Rhizoma and Alismatis Rhizoma.

To reduce microbial loads in medicinal herbs, Cnidii Rhizoma and Alismatis Rhizoma were subjected to electron-beam (e-beam) irradiation at doses (≤10 kGy) as permitted by the Korean Food Code. The effects of e-beam irradiation on the microbial load, stability of the active components, and anti-inflammatory activity of medicinal herbs were determined. We observed that the total aerobic bacteria (TAB; 4.0-7.0 log CFU/g), yeasts and molds (Y&M; 3.3-6.8 log CFU/g), and coliform counts (CC; 3.2-3.8 log CFU/g) in both herb samples were effectively reduced in a dose-dependent manner, resulting in acceptable levels of <3.0 log CFU/g in TAB and Y&M and negative in CC at 10 kGy irradiation. The concentration of the active components (0.87-4.22 mg/g) of Cnidii Rhizoma, including z-ligustilide, chlorogenic acid, senkyunolide A, and ferulic acid, in order of prevalence and those (0.86-2.76 mg/g) of Alismatis Rhizoma, including Alisol B acetate and Alisol B, were not changed at irradiation doses of ≤10 kGy. The extracts of e-beam irradiated Cnidii Rhizoma and Alismatis Rhizoma showed a reduced production of inflammation-related factors, such as nitric oxide, prostaglandin E2, interleukin (IL)-1ß, and IL-6, in a concentration-dependent manner, which was induced by lipopolysaccharide in RAW 264.7 cell. However, there was no significant difference observed at e-beam irradiation doses of 0, 1, 5, and 10 kGy. Thus, we confirm that e-beam irradiation up to 10 kGy was effective for the control of microbial load in Cnidii Rhizoma and Alismatis Rhizoma without causing considerable changes in their major active components and anti-inflammatory activity. The results show the potential of e-beam application for sanitization of medicinal herbs.

Comparison of electronic sensing techniques for screening dried shrimps irradiated using three types of approved radiation with standard analytical methods.

Rapid analytical methods for screening irradiated foods are required to comply with the approved standards for international trade. Dried shrimps irradiated at 1-7 kGy with gamma rays, electron beam (E-beam), and X-rays were screened with an electronic nose (E-nose) and electronic tongue (E-tongue). The data were compared with those from European standard methods (photostimulated luminescence, PSL) and direct epifluorescent filter technique/aerobic plate count, DEFT/APC). All irradiated shrimp samples were clearly discriminated from the non-irradiated control based on PSL photon count measurements and DEFT/APC microbial enumeration. The volatile patterns and taste attributes of the irradiated (>1 kGy from three sources) and control samples could be distinguished by the E-nose and E-tongue analyses through principal component analysis. Verification through electron spin resonance and thermoluminescence analyses validated screening results. The results indicate that E-sensing techniques showed potential for the rapid screening of irradiated foods like dried shrimps.

High glucose alters fetal rat islet transcriptome and induces progeny islet dysfunction.

Offspring of diabetic mothers are susceptible to developing type 2 diabetes due to pancreatic islet dysfunction. However, the initiating molecular pathways leading to offspring pancreatic islet dysfunction are unknown. We hypothesized that maternal hyperglycemia alters offspring pancreatic islet transcriptome and negatively impacts offspring islet function. We employed an infusion model capable of inducing localized hyperglycemia in fetal rats residing in the left uterine horn, thus avoiding other factors involved in programming offspring pancreatic islet health. While maintaining euglycemia in maternal dams and right uterine horn control fetuses, hyperglycemic fetuses in the left uterine horn had higher serum insulin and pancreatic beta cell area. Upon completing infusion from GD20 to 22, RNA sequencing was performed on GD22 islets to identify the hyperglycemia-induced altered gene expression. Ingenuity pathway analysis of the altered transcriptome found that diabetes mellitus and inflammation/cell death pathways were enriched. Interestingly, the downregulated genes modulate more diverse biological processes, which includes responses to stimuli and developmental processes. Next, we performed ex and in vivo studies to evaluate islet cell viability and insulin secretory function in weanling and adult offspring. Pancreatic islets of weanlings exposed to late gestation hyperglycemia had decreased cell viability in basal state and glucose-induced insulin secretion. Lastly, adult offspring exposed to in utero hyperglycemia also exhibited glucose intolerance and insulin secretory dysfunction. Together, our results demonstrate that late gestational hyperglycemia alters the fetal pancreatic islet transcriptome and increases offspring susceptibility to developing pancreatic islet dysfunction.

Optimization of green extraction methods for cinnamic acid and cinnamaldehyde from Cinnamon (Cinnamomum cassia) by response surface methodology.

The major compounds of cinnamon are cinnamic acid and cinnamaldehyde, for which the conditions of microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE), and reflux extraction (RE) were optimized using response surface methodology for comparing their efficiencies in terms of extraction yield, consumption of time and energy, and CO2 emission. The results indicated MAE superiority to UAE and RE owing to the highest yield of target compounds (total yield: 0.89%, cinnamic acid: 6.48 mg/100 mL, and cinnamaldehyde: 244.45 mg/100 mL) at optimum MAE conditions: 59% ethanol, 147.5 W microwave power and 3.4 min of extraction time. RE resulted in comparable yields with the highest consumption of time, energy, and solvent, and least CO2 emission. Therefore, it is concluded that MAE is the most efficient method for green extraction of cinnamic acid and cinnamaldehyde from cinnamon powder compared to UAE and RE.

Assessment of microbial contaminations in commercial frozen duck meats and the application of electron beam irradiation to improve their hygienic quality.

BACKGROUND: High microbial load is a serious concern in terms of the health-related safety of products of animal origin. In this study, the microbial loads of commercial frozen duck-meat products, including bone-in whole raw, boneless sliced raw, and boneless whole smoked, were investigated for pathogenic contamination. The application of electron beam irradiation was also investigated. RESULTS: The samples revealed a serious microbial threat (102 -105 CFU g-1 for total aerobic bacteria and positive for foodborne pathogens), which required effective decontamination technology. Electron-beam irradiation (0, 1, 3, and 7 kGy) could potentially improve the hygienic quality of duck-meat samples. The D10 values for Listeria monocytogenes and Salmonella Typhi were 0.47 and 0.51 kGy, respectively. A direct epifluorescent filter technique and aerobic plate count (DEFT/APC) method was used for screening, while electron-spin resonance (ESR) spectroscopy and gas chromatography with mass spectrometry were effective as confirmatory techniques to identify radiation-induced markers in frozen duck meat. CONCLUSION: Electron-beam irradiation has the potential to ensure the microbial safety and hygienic quality of commercial duck meats. Identification of the samples for their irradiation history was also possible using radiation-induced detection markers, including the DEFT/APC, hydroxyapatite ESR radicals, and hydrocarbons. © 2018 Society of Chemical Industry.

Assessment of Microbial and Radioactive Contaminations in Korean Cold Duck Meats and Electron-Beam Application for Quality Improvement.

Animal-origin food products pose serious threat to public food safety due to high microbial loads. The microbial and radioactive contaminations in commercial cold duck meat products were evaluated. Ten different lots of commercial samples (C1-C10) were classified based on type and smoking process. All samples were highly contaminated (< 4-7 Log CFU/g) with total aerobic bacteria (TAB), yeasts and molds (Y&M), and 7 samples (C1-C7) were positive for coliforms. Furthermore, three samples were contaminated with Listeria monocytogenes (C4-C6) and one with Salmonella typhimurium (C6). No radionuclides (131I, 137Cs, and 134Cs) were detected in any sample. The results of DEFT (direct epifluorescent filter technique)/APC (aerobic plate count), employed to screen pre-pasteurization treatments of products, indicated that smoked samples were positive showing DEFT/APC ratios higher than 4. Notably, the samples showed a serious threat to microbial safety, thus were irradiated with electron-beam (e-beam). The D10 values for S. typhimurium and L. monocytogenes were 0.65 and 0.42 kGy, respectively. E-beam application at 3 and 7 kGy resulted in reduction of initial TAB, Y&M, and coliform populations by 3 and 6 log cycles, respectively. Thus, e-beam was proven to be a good decontamination approach to improve the hygiene of cold duck meat.

Physicochemical Qualities and Flavor Patterns of Traditional Chinese Vinegars Manufactured by Different Fermentation Methods and Aging Periods.

Physicochemical properties of Fujian Yongchun aged vinegar (FYAV) and Shanxi mature vinegar (SMV) were compared in terms of the fermentation methods applied and aging periods (3, 5, 8, and 10 years), and combined E-nose/E-tongue analyses were performed to assess their flavors. Compared with submerged fermentation-derived FYAV, solid-state fermentation-derived SMV showed higher values of pH, brix, soluble solids, total phenolic content, and antioxidant activity, but not total acidity or total organic acids. Aging period resulted in an increase in pH, total phenolic content, and antioxidant activity. Principal component analysis based on E-tongue/E-nose analyses was performed to distinguish between the vinegars produced by different fermentation methods and under aging periods. Solid-state fermentation and an aging process were considered good techniques for vinegar brewing, considering the various organic acids and high levels of total phenolics and antioxidant activity.

Optimization of microwave-assisted extraction of total extract, stevioside and rebaudioside-A from Stevia rebaudiana (Bertoni) leaves, using response surface methodology (RSM) and artificial neural network (ANN) modelling.

Stevia rebaudiana (Bertoni) consists of stevioside and rebaudioside-A (Reb-A). We compared response surface methodology (RSM) and artificial neural network (ANN) modelling for their estimation and predictive capabilities in building effective models with maximum responses. A 5-level 3-factor central composite design was used to optimize microwave-assisted extraction (MAE) to obtain maximum yield of target responses as a function of extraction time (X1: 1-5min), ethanol concentration, (X2: 0-100%) and microwave power (X3: 40-200W). Maximum values of the three output parameters: 7.67% total extract yield, 19.58mg/g stevioside yield, and 15.3mg/g Reb-A yield, were obtained under optimum extraction conditions of 4min X1, 75% X2, and 160W X3. The ANN model demonstrated higher efficiency than did the RSM model. Hence, RSM can demonstrate interaction effects of inherent MAE parameters on target responses, whereas ANN can reliably model the MAE process with better predictive and estimation capabilities.

E-Beam Irradiation for Improving the Microbiological Quality of Smoked Duck Meat with Minimum Effects on Physicochemical Properties During Storage.

This study was performed to evaluate the effect of different doses (0, 1.5, 3, and 4.5 kGy) of e-beam irradiation on the quality parameters (pH, Hunter's parameter, and heme pigment) and stability qualifiers (peroxide value [POV], thiobarbituric acid reactive substances [TBARSs], and total volatile basic nitrogen [TVBN]) of smoked duck meat during 40 d of storage under vacuum packaging at 4 °C. The initial populations of total bacteria (7.81 log CFU/g) and coliforms (5.68 log CFU/g) were reduced by approximately 2 to 5 log cycles with respect to irradiation doses. The results showed that pH, myoglobin, met-myoglobin, L* , a* , and b* showed significant differences with respect to different doses and storage intervals; a* and b* did not vary significantly because of storage. Higher pH was found in samples treated with 4.5 kGy at 40 d, while the minimum was observed in nonirradiated samples at day 0 of storage. Higher POV (2.31 ± 0.03 meq peroxide/kg) and TBARS (5.24 ± 0.03 mg MDA/kg) values were found in 4.5 kGy-treated smoked meat at 40 d and the lowest was reported in 0 kGy-treated meat at initiation of storage (0 d). However, irradiation suppressed TVBN during storage and higher TVBN (7.09 ± 0.32 mg/100 mL) was found in duck meat treated with 0 kGy at 40 d. The electronic nose (e-nose) effectively distinguished flavor profiles during the different storage intervals. The results showed that different sensory attributes did not vary significantly with respect to the dose of irradiation. We conclude that low dose of e-beam irradiation and vacuum packaging is beneficial for safety and shelf life extension without affecting the sensory characteristics of smoked duck meat.

Accelerated hydration reaction of an unsymmetrical tolan evidenced by a Hg(ii)-trapped macrocycle and its application as a Hg(ii)-selective indicator.

Hg(ii)-mediated hydration reactions of unsymmetrical quinoline type tolans were studied. The observed accelerated reactions of the tolans rely on the additional binding motifs of the tolan, as supported by the X-ray structure of the macrocycle (2b). The analyte-specific reaction allows us to detect Hg(ii) in buffered media.

Application of E-tongue, E-nose, and MS-E-nose for discriminating aged vinegars based on taste and aroma profiles.

The combined approach of E-senses was undertaken to discriminate the taste and aroma profiles of aged vinegars, including Chinese Shanxi extra aged vinegar (EX, 5-8 years), Japanese black vinegar (BL, 1-5 years), and Italian balsamic vinegar (BA, 8-25 years). The principal component analysis plot for E-tongue revealed that EX, BL, and BA formed individual groups with clear distinctions. The volatile compounds detected in E-nose based on Kovats indices represented that a major component in all vinegars was matched to acetic acid, which showed a strong relationship with the aging period (R=0.8186-0.9998). The discriminant function analysis plot accompanied by the MS-E-nose indicated that more volatiles were produced during prolonged aging in cereal vinegar, whereas the aging process degraded volatile constituents in fruit vinegar. Categorization with respect to aging years was possible using both E-noses. The approach using E-senses indicated a better discrimination for distinguishing the complex sensory information in aged vinegars.

Hypoxia induces cyclophilin B through the activation of transcription factor 6 in gastric adenocarcinoma cells.

Hypoxia is an important form of physiological stress that induces cell death, due to the resulting endoplasmic reticulum (ER) stress, particularly in solid tumors. Although previous studies have indicated that cyclophilin B (CypB) plays a role in ER stress, there is currently no direct information supporting the mechanism of CypB involvement under hypoxic conditions. However, it has previously been demonstrated that ER stress positively regulates the expression of CypB. In the present study, it was demonstrated that CypB is transcriptionally regulated by hypoxia-mediated activation of transcription factor 6 (ATF6), an ER stress transcription factor. Subsequently, the effects of ATF6 on CypB promoter activity were investigated and an ATF6-responsive region in the promoter was identified. Hypoxia and ATF6 expression each increased CypB promoter activity. Collectively, these results demonstrate that ATF6 positively regulates the expression of CypB by binding to an ATF6-responsive region in the promoter, which may play an important role in the attenuation of apoptosis in the adaption to hypoxia. These results suggest that CypB may be a key molecule in the adaptation of cells to hypoxic conditions.

Bis-ureidoquinoline as a selective fluoride anion sensor through hydrogen-bond interactions.

Bis-ureidoquinoline shows a characteristic UV-vis absorbance and turn-on fluorescence changes in the presence of the fluoride anion. Such selective changes probably originate from the hydrogen-bond interactions, as shown by the (1)H NMR titration and DFT calculations. Bis-ureidoquinoline can be used as a fluoride-selective sensor for the detection of fluoride anions under illumination from a laboratory hand-held UV lamp.