Effects of HA and NA glycosylation pattern changes on the transmission of avian influenza A(H7N9) virus in guinea pigs

AbstractAvian influenza H7N9 virus has posed a concern of potential human-to-human transmission by resulting in seasonal virus-like human infection cases. To address the issue of sustained human infection with the H7N9 virus, here we investigated the effects of hemagglutinin (HA) and neuraminidase (NA) N-linked glycosylation (NLG) patterns on influenza virus transmission in a guinea pig model. Based on the NLG signatures identified in the HA and NA genetic sequences of H7N9 viruses, we generated NLG mutant viruses using either HA or NA gene of a H7N9 virus, A/Anhui/01/2013, by reverse genetics on the 2009 pandemic H1N1 virus backbone. For the H7 HA NLG mutant viruses, NLG pattern changes appeared to reduce viral transmissibility in guinea pigs. Intriguingly, however, the NLG changes in the N9 NA protein, such as a removal from residue 42 or 66 or an addition at residue 266, increased transmissibility of the mutant viruses by more than 33%, 50%, and 16%, respectively, compared with a parental N9 virus. Given the effects of HA-NA NLG changes with regard to viral transmission, we then generated the HA-NA NLG mutant viruses harboring the H7 HA of double NLG addition and the N9 NA of various NLG patterns. As seen in the HA NLG mutants above, the double NLG-added H7 HA decreased viral transmissibility. However, when the NA NLG changes occurred by a removal of residue 66 and an addition at 266 were additionally accompanied, the HA-NA NLG mutant virus recovered the transmissibility of its parental virus. These demonstrate the effects of specific HA-NA NLG changes on the H7N9 virus transmission by highlighting the importance of a HA-NA functional balance

Caloric restriction of db/db mice reverts hepatic steatosis and body weight with divergent hepatic metabolism

Non-alcoholic fatty liver disease (NAFLD) is one of the most frequent causes of liver disease and its prevalence is a serious and growing clinical problem. Caloric restriction (CR) is commonly recommended for improvement of obesity-related diseases such as NAFLD. However, the effects of CR on hepatic metabolism remain unknown. We investigated the effects of CR on metabolic dysfunction in the liver of obese diabetic db/db mice. We found that CR of db/db mice reverted insulin resistance, hepatic steatosis, body weight and adiposity to those of db/m mice. H-NMR- and UPLC-QTOF-MS-based metabolite profiling data showed significant metabolic alterations related to lipogenesis, ketogenesis, and inflammation in db/db mice. Moreover, western blot analysis showed that lipogenesis pathway enzymes in the liver of db/db mice were reduced by CR. In addition, CR reversed ketogenesis pathway enzymes and the enhanced autophagy, mitochondrial biogenesis, collagen deposition and endoplasmic reticulum stress in db/db mice. In particular, hepatic inflammation-related proteins including lipocalin-2 in db/db mice were attenuated by CR. Hepatic metabolomic studies yielded multiple pathological mechanisms of NAFLD. Also, these findings showed that CR has a therapeutic effect by attenuating the deleterious effects of obesity and diabetes-induced multiple complications

Immunogenicity of influenza vaccination in patients with cancer receiving immune checkpoint inhibitors

Among prospectively enrolled adult patients with cancer receiving immune checkpoint inhibitors (ICIs; n = 46) or cytotoxic agents (n = 90), seroprotection and seroconversion rates after seasonal quadrivalent influenza vaccinations were higher with ICI than with cytotoxic chemotherapy. These results support annual influenza vaccinations for cancer patients receiving ICIs.

Metabolic Profiles, Bioactive Compounds, and Antioxidant Capacity in Lentinula edodes Cultivated on Log versus Sawdust Substrates

Lentinula edodes (shiitake) is a popular nutritious edible mushroom with a desirable aroma and flavor. Traditional cultivation of L. edodes on beds of logs has been replaced by cultivation on sawdust, but the effects of cultivation changes on L. edodes mushrooms have not been well characterized. We determined the metabolic profile, bioactive compounds, and antioxidant capacity in L. edodes grown on log or sawdust substrates. Metabolic profiles of L. edodes extracts were determined by 1H nuclear magnetic resonance (NMR) and ultra-performance liquid chromatography to quadrupole time-of-flight mass spectrometry. Principal component analysis score plots from 1H NMR analysis showed clear differences between samples. Concentrations of primary metabolites, especially amino acids, generally decreased in L. edodes grown on logs compared to sawdust. Phenolic compounds showed variations in concentration depending on the cultivation method. Bioactive compounds and their antioxidant capacity were analyzed spectrophotometrically. L. edodes cultivated on logs had high concentrations of bioactive compounds with strong antioxidant capacity compared to L. edodes cultivated on sawdust. Thus, the concentration of primary metabolites was high in L. edodes grown on sawdust, which produces a high growth rate. In contrast, log-cultivated L. edodes, which were similar to wild mushrooms, had high levels of bioactive compounds and high antioxidant capacity. This information is useful for determining optimal cultivation conditions for nutritional and medicinal uses of L. edodes mushrooms

Metabolite Profiling of Chestnut (Castanea crenata) According to Origin and Harvest Time Using 1H NMR Spectroscopy

Chestnuts are an important food crop commonly used as a food ingredient due to their nutritional properties and potential health benefits. In Korea, chestnuts have been crossbred to develop cultivars with insect resistance and high productivity, producing multiple chestnut varieties. This study classified 17 Castanea crenata cultivars produced in Korea according to origin and harvest time and determined the metabolites in chestnut kernels using 1H nuclear magnetic resonance spectroscopy. The 17 C. crenata cultivars were divided into four groups based on their geographic origin: Korean native, Korean hybrid, Japanese native, and Japanese hybrid. The cultivars were also divided into three groups depending on their harvest period: early-ripening cultivar, mid-ripening cultivar, and late-ripening cultivar. The partial least squares-discriminant analysis score plot revealed differences among the groups. Identified metabolites, including amino acids, organic acids, and sugars, contributed to discriminating the origin and harvest time of the C. crenata chestnut kernels. Significant differences were observed, mainly in amino acids, which suggests that the composition of amino acids is one factor influenced by both the origin and harvest time of C. crenata. These results are useful to both growers and breeders because they identify the nutritional and metabolic characteristics of each C. crenata cultivar

UPLC-QTOF-MS-Based Metabolomics and Antioxidant Capacity of <i>Codonopsis lanceolata</i> from Different Geographical Origins

Codonopsis lanceolata (C. lanceolata) has been commonly utilized as a therapeutic plant in traditional medicine. In this study, we examined variations in metabolites in C. lanceolata roots grown in different regions using ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). Multivariate analysis showed that the metabolite profiles of plants grown in Hoengseong and Jeongseon were more similar to each other than to that of C. lanceolata grown in Jeju. Most primary metabolites were present at higher levels in C. lanceolata grown in Jeju. In contrast, C. lanceolata grown in Hoengseong and Jeongseon had high levels of secondary metabolites such as phenylpropanoids and triterpenoid saponins, respectively. In addition, the bioactive compound content and antioxidant capacity of in C. lanceolata grown in Hoengseong and Jeongseon were observed to be higher than those of C. lanceolata grown in Jeju. This study suggests that metabolomics is an effective approach to investigate the difference of metabolite profiling in C. lanceolata from different geographical origins, and is useful for evaluating its pharmacological potential

Metabolic Profiling of Chestnut Shell (Castanea crenata) Cultivars Using UPLC-QTOF-MS and Their Antioxidant Capacity

The inner shell of the chestnut (Castanea crenata) has long been used in Asia as a medicinal herb for improving digestion and blood circulation, and treating diarrhea. However, most chestnut shells are now treated as waste materials in industrial peeling processes. In this study, we examined the metabolite variation among major cultivars of C. crenata shells using mass spectrometry. Among five representative cultivars, Okkwang, Porotan, and Ishizuuchi had higher levels of bioactive compounds, such as ellagic acid derivatives, ellagitannins, flavonoids, and gallic acid derivatives. Their antioxidant capacity was positively correlated with their chemical composition. The byproducts (whole shells) from the industrial peeling process were re-evaluated in comparison with the inner shell, a rich source of phenolic compounds. The phenolic acids and flavonoid glucoside derivatives were significantly higher in the whole shells, whereas the levels of flavonoids were higher in the inner shells. In addition, the whole shell extracts significantly reduced cellular reactive oxygen species production compared to the inner shell extracts. This study demonstrated the different biochemical benefits of different C. crenata cultivars through metabolic profiling and suggests that the whole shell could be used as a functional ingredient, as it has the highest levels of bioactive products and antioxidant effects

LC/MS-Based Polar Metabolite Profiling Identified Unique Biomarker Signatures for Cervical Cancer and Cervical Intraepithelial Neoplasia Using Global and Targeted Metabolomics

Cervical cancer remains one of the most prevalent cancers among females worldwide. Therefore, it is important to discover new biomarkers for early diagnosis of cervical intraepithelial neoplasia (CIN) and cervical cancer, preferably non-invasive ones. In the present study, we aimed to identify unique metabolic signatures for CINs and cervical cancers using global and targeted metabolomic profiling. Plasma samples (69 normal, 55 CIN1, 42 CIN2/3, and 60 cervical cancer) were examined by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-QTOF-MS) coupled with multivariate statistical analysis. Metabolic pathways were analyzed using the integrated web-based tool MetaboAnalyst. A multivariate logistic regression analysis was conducted to evaluate the combined association of metabolites and human papillomavirus (HPV) status with the risk of cervical carcinogenesis. A total of 28 metabolites exhibiting discriminating levels among normal, CIN, and cervical cancer patients (Kruskal&#8211;Wallis test p &lt; 0.05) were identified in the global profiling analysis. The pathway analysis showed significantly altered alanine, aspartate, and glutamate metabolic pathways (FDR p-value &lt; 0.05) in both the discovery and validation phases. Seven metabolites (AMP, aspartate, glutamate, hypoxanthine, lactate, proline, and pyroglutamate) were discriminated between CINs and cervical cancer versus normal (area under the curve (AUC) value &gt; 0.8). The levels of these metabolites were significantly high in patients versus normal (p &lt; 0.0001) and were associated with increased risk of developing CIN2/3 and cervical cancer. Additionally, elevated levels of the seven metabolites combined with positive HPV status were correlated with substantial risk of cancer progression. These results demonstrated that metabolomics profiling is capable of distinguishing CINs and cervical cancers from normal and highlighted potential biomarkers for the early detection of cervical carcinogenesis

Enantioselective Cyclopropanation with α‑Alkyl-α-diazoesters Catalyzed by Chiral Oxazaborolidinium Ion: Total Synthesis of (+)-Hamavellone B

Chiral oxazaborolidinium ion-catalyzed asymmetric cyclopropanation of α- or α,β-substituted acroleins with α-alkyl-α-diazoesters has been developed. With this methodology, chiral functionalized cyclopropanes containing a quaternary stereogenic center were obtained with high to excellent enantioselectivities (up to >99% ee). The synthetic utility of optically enriched functionalized cyclopropane was demonstrated in the first total synthesis of (+)-hamavellone B, which establishes the absolute configuration of natural (+)-hamavellone B

Rational Molecular Design for Non-aqueous Atomic Layer Deposition of Zinc Oxide

Zinc oxide (ZnO) is a transparent wide band gap semiconductor material with various possible applications in form of thin films. Most previous studies on atomic layer deposition (ALD) of ZnO thin films utilized a few well-known processes with diethylzinc (DEZ) and counter-reactants such as H2O and O3. However, O3 and H2O reactants have relatively strong reactivity, so that they are not suitable for substrates sensitive to oxidation. Therefore, development of milder non-aqueous alternative ALD process for ZnO is highly desired. In this study, we introduce ALD of ZnO using alcohols with theoretically optimized molecular structure. To discover suitable alcohol reactants for ZnO ALD, reaction pathways and reactivity between various types of alcohol reactants with surface-ethyl groups were evaluated through density functional theory calculations. It was found that unsaturated allylic alcohols would have the lowest activation energy for ZnO ALD via an allylic rearrangement mechanism. Experimental, novel ALD processes for ZnO using DEZ with alcohol as oxygen sources are set up. Ethanol as a typical simple alcohol is compared to 2-methyl-3-buten-2-ol (MBO) as an alcohol with optimal molecular structure setup. ALD ZnO films using MBO showed processes and material properties comparable to those of H2O-ALD ZnO. ZnO thin films as transparent conducting oxide could be obtained, and device performances of thin-film transistors based on alcohol-ALD processes are evaluated