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Neuro-endocrine prostate cancer (NEPC) accounts for about 20% of lethal metastatic castration-resistant prostate cancer (CRPC). NEPC has the most aggressive biologic behavior of all prostate cancers and is associated with poor patient outcome. Effective treatment for NEPC is not available because NEPC exhibit distinct cell-surface expression profiles compared to other types of prostate cancer. Recently, the carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5) (known as CEA or CD66e) was suggested to be a specific surface protein marker for NEPC. Therefore, we identified a new, fully-human anti-CEACAM5 monoclonal antibody, 1G9, which bound to the most proximal membrane domains, A3 and B3, of CEACAM5 with high affinity and specificity. It shows no off-target binding to other CEACAM family members, membrane distal domains of CEACAM5, or 5800 human membrane proteins. IgG1 1G9 exhibited CEACAM5-specific ADCC activity toward CEACAM5-positive prostate cancer cells in vitro and in vivo. Chimeric antigen receptor T cells (CAR-T) based on scFv 1G9 induced specific and strong antitumor activity in a mouse model of prostate cancer. Our results suggest that IgG1 and CAR-T cells based on 1G9 are promising candidate therapeutics for CEACAM5-positive NEPC and other cancers.

Rechargeable lithium-ion batteries (LIBs) have a wide range of applications but face challenges in harsh working or operating environments at high temperatures. In this work, a solid polymer electrolyte with MWCNT-COOH as an additive (MWCNT-SPE) was obtained. MWCNT-SPE has a high thermal stability and can be used in high-temperature operating environments. Solid-state lithium batteries based on MWCNT-SPE and LiFePO4 were assembled. The resulting lithium batteries exhibited excellent electrochemical properties at 70 and 120 °C, demonstrating a wide range of operations suitable for solid-state batteries with extreme demands. The symmetrical Li/MWCNT-SPE/Li cell operated for 1800 h with low polarization voltage and no short circuit, and the LiFePO4/MWCNT-SPE/Li cell delivered superior cycling performance under both 0.2 and 0.5 C-rates, indicating that the interface compatibility between the lithium metal and MWCNT-SPE membrane was good and could effectively suppress the formation of lithium dendrites. The superior performance of the resulting MWCNT-SPE was due to the weak interaction between PEO, PVDF-HFP, and MWCNT-COOH, which reduced the tendency of PEO's crystallinity and thereby significantly increased the Li+ migration ability and improved the cycling life of the batteries.

Hypercholesterolemia is the main cause of cardiovascular disease worldwide, and the regulation of cholesterol homeostasis is essential for human health. Lactobacillus is present in large quantities in the human intestine. As the normal flora in the gut, lactobacillus plays an important role in regulating metabolism in the human body. Lactobacillus can regulate the cholesterol content by regulating the expression of genes involved in cholesterol synthesis, metabolism, and absorption. This article reviews the biological effects and mechanisms of lactobacillus that mediate the expression of NPC1L1, CYP7A1, ABCG5, ABCG8, and other genes to inhibit cholesterol absorption, and discusses the mechanism of reducing cholesterol by lactobacillus in cells in vitro, to provide a theoretical basis for the development and utilization of lactobacillus resources.

The ages of Carlin-type gold deposits in the Golden Triangle of South China have long been questioned due to the general lack of minerals unequivocally linked to gold deposition that can be precisely dated using conventional radiogenic isotope techniques. Recent advances in U-Pb methods show that calcite can be used to constrain the ages of hydrothermal processes, but few studies have been applied to ore deposits. Herein, we show that this approach can be used to constrain the timing of hydrothermal activity that generated and overprinted the giant Shuiyindong Carlin-type gold deposit in the Golden Triangle. Three stages of calcite (Cal-1, Cal-2, and Cal-3) have been recognized in this deposit based on crosscutting relationships, cathodoluminescence colors, and chemical (U, Pb, and rare earth element [REE]) and isotope (C, O, Sr) compositions. Cal-1 is texturally associated with ore-stage jasperoid and disseminated Au-bearing arsenian pyrite in hydrothermally altered carbonate rocks, which suggests it is synmineralization. Cal-2 fills open spaces and has a distinct orange cathodoluminescence, suggesting that it precipitated during a second fluid pulse. Cal-1 and Cal-2 have similar carbonate rock-buffered chemical and isotopic compositions. Cal-3 occurs in veins that often contain realgar and/or orpiment and are chemically (low U, Pb, and REE) and isotopically (higher δ13C, lower δ18O and Sri values) distinct from Cal-1 and Cal-2, suggesting that it formed from a third fluid. U-Pb isotope analyses, by laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) for U-rich Cal-1 and Cal-2 and by LA-multicollector (MC)-ICP-MS for U-poor Cal-3, yield well-defined age constraints of 204.3 to 202.6, 191.9, and 139.3 to 137.1 Ma for Cal-1, Cal-2, and Cal-3, respectively. These new ages suggest that the Shuiyindong gold deposit formed in the late Triassic and was overprinted by hydrothermal events in the early Jurassic and early Cretaceous. Given the association of Cal-3 with orpiment and realgar, and previous geochronologic studies of several other major gold deposits in the Golden Triangle, we infer that the latest stage of calcite may be associated with an early Cretaceous regional gold metallogenic event. Combined with existing isotopic ages in the region, these new ages lead us to propose that Carlin-type gold deposits in the Golden Triangle formed during two metallogenic episodes in extensional settings, associated with the late Triassic Indochina orogeny and early Cretaceous paleo-Pacific plate subduction. This study shows that the calcite U-Pb method can be used to constrain the timing of Carlin-type gold deposits and successive hydrothermal events.

Nonalcoholic fatty liver disease (NAFLD) is characterized by high morbidity. Although long noncoding RNAs (lncRNAs) are known to have a role in NAFLD pathogenesis, the identified lncRNA types are limited. In this study, NAFLD models were established in vitro and in vivo using free fatty acid-treated LO2 cells and high-fat diet-fed mice, respectively. Microarray data were downloaded from the Gene Expression Omnibus database, and AC012668 was selected for further analysis. Cell viability and apoptosis were measured using Cell Counting Kit 8 and flow cytometry assays. RNA expression was detected using reverse transcription-quantitative polymerase chain reaction. Triglyceride (TG) content and lipid deposition were detected using enzyme-linked immunosorbent assay and Oil-Red O staining. Western blotting was used to visualize protein expression. Starbase and TargetScan were used to predict the target miRNA and gene, and the predictions were verified through RNA pull-down and luciferase reporter assays. AC012668 expression levels were significantly suppressed in NAFLD models, whereas AC012668 overexpression inhibited lipogenesis-related gene (SCD1, SREBP1, FAS) expression and TG/lipid accumulation in vitro. Subsequently, miR-380-5p was predicted and verified to target AC012668, and its expression was notably increased in the NAFLD cell model. Moreover, transfection of miR-380-5p antagonized the effects of AC012668 on lipid formation and accumulation. LRP2 was confirmed to be the target gene of miR-380-5p and was downregulated in the NAFLD cell model. Silencing LRP2 reversed the effects of the miR-380-5p inhibitor on lipid formation and accumulation. AC012668 inhibited NAFLD progression via the miR-380-5p/LRP2 axis. These findings may provide a novel strategy against NAFLD.

Payenapara lleloneura Kurz. (Kan-zaw), an endemic medicinal plant only found in Tanintharyi Region of Myanmar, is widely used in the treatment of various cancer and different ailments. In the present research, the seeds were phytochemical investigated for their nutritional potential for their use as functional foods or novel diet oil resources. Nutritional evaluation showed that the seeds are rich in fats and carbohydrates (soluble sugars and starch). Fatty acid analyses showed that the seeds accumulate very rich α-eleostearic acid (α-ESA, 18:3Δ9cis,11trans,13trans), an important conjugated fatty acid, up to more than 70 ​% of total fatty acids. The seed oil derived from the Kan-zaw tree contains approximately 3.25 ​% β-eleostearic acid (18:3Δ9trans,11trans,13trans), an unusual conjugated fatty acid that imparts a potent anticancer application and industrially important drying qualities to Kan-zaw oil. Physicochemical properties of the Kan-zaw seeds were examined; petroleum ether (60–90 ​°C) extract of seed oils were also investigated for the saponification value, iodine value and estimation of acid value. Further, the present study investigated cytotoxic potential of ethanol, methanol, acetone, chloroform Kan-zaw seed extracts and commercial Kan-zaw oil against human cervical cancer cell line (HeLa). The Kan-zaw extracts and oil have shown significant anticancer activity on HeLa cells.

This study investigates the microstructure, tensile properties, and high-cycle fatigue resistance of twin-roll-cast Mg-3Al-1Zn (wt%) alloy strips with thicknesses of 1 mm, 1.5 mm, and 3 mm. The investigation results reveal that the 1-and 1.5-mm-thick strips show a fully dynamically recrystallized (DRXed) microstructure consisting of fine equiaxed DRXed grains, whereas the 3-mm-thick strip shows a partially DRXed microstructure containing very coarse elongated unDRXed grains because of the insufficient strain imposed during twin-roll casting. The inhomogeneous microstructure of the 3-mm-thick strip leads to a large deviation in its tensile elongation. The average grain size of the strips increases with increasing strip thickness, which results in reductions in both their tensile strength and their ductility because of the weakened grain-boundary hardening effect and the promoted formation of undesirable twins, respectively. The high-cycle fatigue resistance in the stress regime with finite fatigue life is similar for all three strips, but the fatigue strength with infinite fatigue life decreases from 175 MPa to 140 MPa as the strip thickness increases from 1 mm to 3 mm. The fatigue strength (FL) increases linearly with increasing yield strength (YS) according to the relationship FL = -199.5 + 2.03·YS.

Rapidly proliferating cells such as vascular smooth muscle cells (VSMCs) require metabolic programs to support increased energy and biomass production. Thus, targeting glutamine metabolism by inhibiting glutamine transport could be a promising strategy for vascular disorders such as atherosclerosis, stenosis, and restenosis. V-9302, a competitive antagonist targeting the glutamine transporter, has been investigated in the context of cancer; however, its role in VSMCs is unclear. Here, we examined the effects of blocking glutamine transport in fetal bovine serum (FBS)- or platelet-derived growth factor (PDGF)-stimulated VSMCs using V-9302. We found that V-9302 inhibited mTORC1 activity and mitochondrial respiration, thereby suppressing FBS- or PDGF-stimulated proliferation and migration of VSMCs. Moreover, V-9302 attenuated carotid artery ligation-induced neointima in mice. Collectively, the data suggest that targeting glutamine transport using V-9302 is a promising therapeutic strategy to ameliorate occlusive vascular disease.

Sickness symptoms exerted via inflammatory responses occur in several infectious and chronic diseases. A growing body of evidence suggests that altered nutrient availability and metabolism are tightly coupled to inflammatory processes. However, the relationship between metabolic shifts and the development of the sickness response has not been explored fully. Therefore, we aimed to evaluate metabolic phenotypes with a mouse model showing sickness symptoms via systemic administration of lipopolysaccharide (LPS) in the present study. LPS injection elevated the lipid utilization and circulating levels of fatty acids. It also increased the levels of β-hydroxybutyric acid, a ketone body produced from fatty acids. We confirmed the functional connectivity between nutrient utilization and inflammatory responses and demonstrated enhanced lipid utilization in the hypothalamus providing insights into hypothalamic control of sickness responses. Collectively, these findings could help develop new therapeutic strategies to treat patients with severe sickness symptoms associated with infectious and chronic human diseases.

Background:. MicroRNAs are closely associated with the progression and outcomes of multiple human diseases, including sepsis. In this study, we examined the role of miR-23a in septic injury. Methods:. Lipopolysaccharide (LPS) was used to induce sepsis in a rat model and H9C2 and HK-2 cells. miR-23a expression was evaluated in rat myocardial and kidney tissues, as well as H9C2 and HK-2 cells. A miR-23a mimic was introduced into cells to identify the role of miR-23a in cell viability, apoptosis, and the secretion of inflammatory cytokines. Furthermore, the effect of Rho-associated kinase 1 (ROCK1), a miR-23a target, on cell damage was evaluated, and molecules involved in the underlying mechanism were identified. Results:. In the rat model, miR-23a was poorly expressed in myocardial (sham vs. sepsis 1.00 ± 0.06 vs. 0.27 ± 0.03, P

Background To analyse structural characteristics and perifoveal/peripapillary vasculature by OCT in children with peripapillary hyperreflective ovoid mass-like structures (PHOMS) and compare the results with those of normal subjects. Methods Forty-five patients (84 eyes) under 18 years old with blurry disc margin were evaluated with spectral domain-OCT and swept course-OCT. Patients were divided into four groups, according to presence of PHOMS and then the size of the existing PHOMS. Eyes with visible optic disc drusen (ODD) were not included. Foveal avascular zone (FAZ) and vessel densities from macula and optic disc area were assessed and potential associations between vessel density and structural parameters, such as peripapillary retinal nerve fibre layer (pRNFL), and macular ganglion cell and inner plexiform layer (mGCIPL) thickness, were analysed. Results Among 45 patients (eighty-four eyes), coexisting buried ODD were found only in eyes with PHOMS. The scleral canal diameter was significantly smaller in PHOMS positive eyes compared to control eyes. Vessel density measurements from the papillary, peripapillary and optic nerve head (ONH) regions in the large PHOMS group were significantly lower compared to the control group (papillary; P = 0.014, peripapillary; P = 0.001, ONH; P = 0.046). FAZ area and macular vessel densities showed no difference compared to normal eyes in all three PHOMS groups. pRNFL and mGCIPL thickness did not differ among four groups and correlations were also not significant. Conclusions Children with PHOMS have smaller scleral canal and can entail buried ODD. Vessel densities of optic disc area in large PHOMS eyes are significantly lower than in normal eyes.

최근 신종 코로나바이러스 감염증의 확산으로 ‘식(食)문화’에 큰 변화로 인해 O2O 외식 배달 어플리케이션의 사용이 크게 증가하고 있으며, 이용자의 지속이용의도에 대한 분석이 필요한 것으로 판단되었다. 이에 본 연구는 O2O 외식 배달 어플리케이션 이용에 대한 혜택과 희생을 토대로 서비스의 지각된 가치를 인지하고 목표 지향적 행동모형을 적용하여 지속이용의도에 유의미한 영향을 미친다는 구조적인 관계를 조사하고자 하였으며 결과는 다음과 같다. 첫째, 혜택 중 유용성과 즐거움이 지각된 가치에 정(+)의 영향을 주는 것으로 나타났다. 둘째, 희생 중 지각된 비용과 보안 위험성은 지각된 가치에 부(-)의 영향을 주는 것으로 나타났다. 셋째, 지각된 가치는 태도, 주관적 규범, 지각된 행동통제에 정(+)의 영향을 미치는 것으로 나타났다. 넷째, 태도와 주관적 규범은 열망에 정(+)의 영향을 미치는 것으로 나타났다. 다섯째, 열망은 지속이용의도에 정(+)의 영향을 미치는 것으로 나타났으며 마지막으로 지각된 행동통제는 지속이용의도에 정(+)의 영향을 미치는 것으로 나타났다. 본 연구는 O2O 외식 배달 어플리케이션 지속이용의도에 관한 구조적 관계를 혜택과 희생을 바탕으로 목표 지향적 행동모형에 적용했다는 점에서 큰 의의가 있으며, 향후 O2O 외식 배달 어플리케이션 업체의 전략 수립 시 많은 도움이 될 것으로 사료된다.

Amylosucrase (ASase, E.C. 2.4.1.4) is a powerful transglycosylation enzyme that can transfer glucose from sucrose to the hydroxyl (-OH) group of various compounds. In this study, recombinant ASases from Deinococcus geothermalis (DgAS) and Bifidobacterium thermophilum (BtAS) were used to synthesize biosurfactants based on the computational analysis of predicted docking simulations. Successful predictions of the binding affinities, conformations, and three-dimensional structures of three surfactants were computed from receptor-ligand binding modes. DgAS and BtAS were effective in the synthesis of biosurfactants from glyceryl caprylate, glyceryl caprate, and polyglyceryl-2 caprate. The results of the transglycosylation reaction were consistent for both ASases, with glyceryl caprylate acceptor showing the highest concentration, as confirmed by thin layer chromatography. Furthermore, the transglycosylation reactions of DgAS were more effective than those of BtAS. Among the three substrates, glyceryl caprylate glycoside and glyceryl caprate glycoside were successfully purified by liquid chromatography–mass spectrometry (LC–MS) with the corresponding molecular weights.

The source and evolution of ore-forming fluids is important to understand the genesis of Carlin-type gold deposit. Constraints on the source and evolution of ore fluid components by the conventional geochemical methods have long been a challenge due to the very fine-grained nature and complex textures of hydrothermal minerals in these deposits. In this study, we present the crush-leach analyzed solute data of fluid inclusion extracts within quartz, calcite, realgar, and fluorite from the Shuiyindong, Nibao, and Yata gold deposits in the Youjiang Basin, providing new insights into the source and evolution of ore-forming fluids. The results show that the high molar Cl/Br ratios up to 2 508 in fluid inclusion extracts are indicative of a contribution of magmatic hydrothermal fluids. Fluids mixing between basinal and magmatic-hydrothermal fluids are evident on the plots of Cl/Br versus Na/K ratios, showing that ore-stage milky quartz near the magmatic-hydrothermal fluids reflects magma origin of the ore-forming fluids, whereas late ore-stage drusy quartz and realgar near the defined basinal fluids suggest the later input of basinal fluids in late-ore stage. Although the predominately host rocks in Shuiyindong, Nibao and Yata gold deposit are bioclastic limestone, sedimentary tuff, and calcareous siltstone, respectively, the solute data of fluid inclusion extracts records they underwent the similar fluid-rocks reactions between the Na-rich magmatic hydrothermal fluids and the Ca- and Mg-rich host rocks. This study highlights the solute data of fluid inclusion extracts obtained by crush-leach analyses have the potential to fingerprint the source and evolution of ore-forming fluids of the Carlin-type gold deposit.

The development of ceramic-polymer electrolyte with high mechanical and electrochemical properties is the key to address the safety issue of high-performance lithium batteries due to the use of flammable organic liquid electrolytes. In this work, we designed and synthesized a flexible low liquid leakage and high wettability ceramic-polymer electrolyte (CPE) based on poly(vinylidene fluoride) (PVDF)-poly(ethylene oxide) (PEO)-γ-Al2O3 membrane with excellent mechanical and thermal stability. After being wetted by a very small amount of LiPF6-based liquid electrolyte, symmetrical Li, Li/LiFePO4 (LFP), and LFP/Li4Ti5O12 (LTO) cells with this PVDF/PEO-γ-Al2O3-LiPF6 CPE displayed much better cycling stabilities than that of commercial Celgard 2320 separator with a sufficient amount of liquid electrolyte. The CPE requires only a small amount of liquid electrolyte to achieve the desired electrochemical performance of lithium batteries. Its low liquid leakage characteristic also enhances the safety of lithium batteries, with the potential to replace the commercial separators.

The design and synthesis of novel energetic materials with high energy and acceptable sensitivity is significant for the military and civil fields. In this study, a methylene-bridged asymmetric heterocyclic explosive based on strong intermolecular interactions, namely N-(1-((5-(nitroimine)-1,2,4-oxadiazol-3-yl) methyl)-1H-tetrazol-5-yl)nitroimine (4), was designed and synthesized. Its ammonium salt (5) and potassium coordination polymer (6) were also synthesized. The structures of compounds 4, 5 and 6 were determined by single crystal X-ray diffraction analyses. Independent gradient model (IGM) analyses and Hirshfeld surfaces (HS) analyses combined with single crystal analyses revealed the existence of intermolecular interactions in the three compounds. The strong intermolecular hydrogen-bonding, π⋯π, and p⋯π interactions enhance the molecular packing efficiency of 4, leading to high density and acceptable sensitivity. Detonation properties calculated with the professional software package EXPLO5 showed that the new metal-free explosive 4 exhibits superior detonation performance (D = 9296.8 m s−1; P = 38.3 GPa) which is comparable to that of HMX.

To investigate the effects of the activator of AMPK and high glucose on the differentiation of mouse SMSCs, primary SMSCs were isolated from mouse extensor digitorum longus muscle and grown to near confluence (80%). Postconfluent cells were cultured in a growth medium with different inductors: AICAR, glucose, and AICAR mixed with glucose. The specific protein expressions of SMSCs, myoblasts, adipocytes, and brown adipocytes were analyzed on days 0, 3, 5, 7, and 10. The results showed treatment with AICAR in SMSCs markedly activated AMPK phosphorylation (p, To investigate the effects of the activator of AMPK and high glucose on the differentiation of mouse SMSCs, primary SMSCs were isolated from mouse extensor digitorum longus muscle and grown to near confluence (80%). Postconfluent cells were cultured in a growth medium with different inductors: AICAR, glucose, and AICAR mixed with glucose. The present study indicated that the certain energy supplements influence the direction of SMSC differentiation which may contribution on the structure of muscle and meat quality, sequentially.

Muscle fiber is the basic unit of skeletal muscle. Muscle fiber types play a crucial role in improving meat quality. In the current study, Wistar rats were chronic injected with 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR) for four weeks in order to define the 5′-AMP-activated protein kinase (AMPK) protein activity on muscle fiber composition, enzyme activity and the mRNA expression of muscle fiber type-related genes in gastrocnemius (GS) muscle. Food intake and body weight did not show a significant difference after AICAR treatment (P > .05). The AICAR treatment caused a significantly increased number and area proportion of type I muscle fiber and expression level MyHC I mRNA (P

Calcium is the most abundant stored mineral in the human body and is especially vital for bone health; thus, calcium deficiency can cause bone-related diseases, such as osteopenia and osteoporosis. However, a high concentration of serum calcium, which is commonly known as hypercalcemia, can also lead to weakened bones and, in severe cases, osteosarcoma. Therefore, it is necessary to maintain the concentration of calcium that is appropriate for bone biology. In the present study, we aimed to elucidate the effects of high concentration of calcium, approximately 2 folds the normal calcium level, on osteoblast differentiation. The CaCl2 treatment showed dose-dependent suppression of the alkaline phosphatase activity and mineralized nodule formation. Calcium showed cytotoxicity at an extremely high concentration, but a moderately high concentration of calcium that results in inhibitory effects to osteoblast differentiation showed no signs of cytotoxicity. We also confirmed that the CaCl2 treatment repressed the mRNA expression and protein abundance of various osteogenic genes and transcriptional factors. Considered together, these results indicate that a high concentration of calcium negatively regulates the osteoblast differentiation of C2C12 cells.

A novel lipase gene, Lip-1420, was isolated from a metagenomic library constructed from reed marsh from Mt. Jumbong in Korea, comprising 112,500 members of recombinant plasmids. The DNA sequence of Lip- 1420-subclone (5,513 bp) was found to contain at least 11 ORFs according to the GenBank database. The ORF-3 gene was inserted into the pET21a plasmid containing the C-terminal 6-His tag and transformed into E. coli BL21(DE3) to express the recombinant lipase protein. Lip-1420 was purified using a fast protein liquid chromatography system. The gene was registered in GenBank (MH628529). The values of Km and Vmax were determined as 0.268 mM and 1.821 units, respectively, at 40℃ and pH 8.0, using p-nitrophenyl palmitate as the substrate. This lipase belongs to family IV taxonomically because it has conserved HGGG and GDSAG motifs in the constitutive amino acid sequence. According to the predicted structural model, the binding sites are represented by residues H78, G81, D150, S151, A152, V181, and D236. Finally, Lip-1420 showed triolein selectivity for methanolysis between triolein (18:1) and tristearin (18:0) substrates. Further study of the selective mechanism and structure-function relationship of this new lipase could be useful for more practical applications.

Hypoxia, a condition of oxygen deficiency in tissues, features various diseases including solid tumor. Under hypoxia, several reductases such as nitroreductases are elevated. Based on this fact, we have investigated an indirect way to assess the hypoxia susceptibility of different organ tissues (mouse lung, heart, spleen, kidney, and liver) by detecting nitroreductase present within. Among the organs, the kidney showed a notable susceptibility to hypoxia, which was due to the renal medulla, not due to the renal cortex, as observed by ratiometric fluorescence imaging with a probe. The probe features ratiometric signaling, NIR-emitting, two-photon absorbing, and pH-insensitive emission properties, offering a practical tool for studying the nitroreductase activity and, furthermore, hypoxia-associated biological processes.

Resistant starch (RS) is a complex prebiotic carbohydrate beneficial to the human gut. In the present study, four genes encoding for putative amylolytic enzymes, likely to be responsible for RS-degradation, were identified in the genome of Bifidobacterium adolescentis P2P3 by comparative genomic analysis. Our results showed that only three enzymes (RSD1, RSD2, and RSD3) exhibited non-gelatinized high amylose corn starch (HACS)-degrading activity in addition to typical α-amylase activity. These three RS-degrading enzymes (RSD) were composed of multiple domains, including signal peptide, catalytic domain, carbohydrate binding domains, and putative cell wall-anchoring domains. Typical catalytic domains were conserved by exhibiting seven typical conserved regions (I-VII) found mostly in α-amylases. Analysis of enzymatic activity revealed that RSD2 displayed stronger activity toward HACS-granules than RSD1 and RSD3. Comparative genomics in combination with enzymatic experiments confirmed that RSDs might be the key enzymes used by RS-degrading bifidobacteria to degrade RS in a particular ecological niche, such as the human gut.

The cell membrane regulates the flux of materials in and out of cell, cell adhesion, and signaling. Fluorophores that selectively localize on it are in demand for investigations of the molecular events occurring on the outer cell membrane. Commercial membrane trackers based on phospholipids are structurally complex and difficult to modify further. We disclose the zwitterionic (naphthylvinyl)pyridinium dyes that selectively localize on the outer cell membrane and emit blue, green, and red fluorescence, respectively. Notably, they are structurally compact and provide bright fluorescence images of the cell membrane. By comparing with control compounds, we identified minimal structural elements for the "robust" localization of dye on the outer cell membrane. Further, the dyes are two-photon active, enabling high-resolution, deep-tissue imaging. One of the dyes was used to image a spleen tissue, which provided high-resolution fluorescent images with a distinct morphology. In addition, the materials and results disclosed are valuable for the development of membrane-targeting probes and structurally compact fluorophores.

Background Rumen bacteria play a critical role in feed degradation and productivity. This study evaluated the impact of feeding regimen on the rumen microbial populations and fatty acid composition of the meat of sheep. Twenty-four Sunit sheep were raised on a grass pasture from birth to 9 months of age, at which time they were randomly divided into two feeding groups: pasture feeding (PF) and barn feeding (BF). Sheep in the PF group were allowed to graze freely on wild grassland for 3 months. Sheep in the BF group were confined for 3 months to a dry barn, in which they roamed freely with corn straw and corn. Results Sheep in the PF group had greater rumen bacteria diversity. The relative abundances of the genera Butyrivibrio_2, Saccharofermentans and Succiniclasticum were increased, and that of the genus RC9_gut_group was decreased, in the PF compared to the BF sheep. The n-3 polyunsaturated fatty acid contents were greater in meat from PF sheep than from BF sheep. In addition, the α-linolenic acid (C18:3 n-3, ALA) and conjugated linoleic acid (CLA) contents were positively correlated with the abundance of Butyrivibrio_2. Conclusion Grazing may improve the diversity of rumen bacteria and increase the proportion of ALA and CLA in sheep meat. © 2020 Society of Chemical Industry.