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Purpose Phosphatidylinositol Glycan Anchor Biosynthesis, class G (PIGG) is an ethanolamine phosphate transferase catalyzing the modification of glycosylphosphatidylinositol (GPI). GPI serves as an anchor on the cell membrane for surface proteins called GPI-anchored proteins (GPI-APs). Pathogenic variants in genes involved in the biosynthesis of GPI cause inherited GPI deficiency (IGD), which still needs to be further characterized. Methods We describe 22 individuals from 19 unrelated families with biallelic variants in PIGG. We analyzed GPI-AP surface levels on granulocytes and fibroblasts for three and two individuals, respectively. We demonstrated enzymatic activity defects for PIGG variants in vitro in a PIGG/PIGO double knockout system. Results Phenotypic analysis of reported individuals reveals shared PIGG deficiency–associated features. All tested GPI-APs were unchanged on granulocytes whereas CD73 level in fibroblasts was decreased. In addition to classic IGD symptoms such as hypotonia, intellectual disability/developmental delay (ID/DD), and seizures, individuals with PIGG variants of null or severely decreased activity showed cerebellar atrophy, various neurological manifestations, and mitochondrial dysfunction, a feature increasingly recognized in IGDs. Individuals with mildly decreased activity showed autism spectrum disorder. Conclusion This in vitro system is a useful method to validate the pathogenicity of variants in PIGG and to study PIGG physiological functions. Unlabelled Image

In situ g-C3N4@ZnO nanocomposites (with 0, 1, 3, 5, and 7 wt.% of g-C3N4 in nanocomposite) were synthesized via a one-pot hydrothermal method using precursors of urea, zinc nitrate hexahydrate, and hexamethylenetetramine. The g-C3N4@ZnO nanocomposites were characterized by X-ray diffraction, scanning electron microscope, diffuse reflectance spectroscopy, and photoluminescence spectroscopy. The photocatalyst activity of g-C3N4@ZnO nanocomposites was evaluated via methylene blue degradation experiment under visible light irradiation. The g-C3N4@ZnO nanocomposites showed an enhancement in photocatalytic activity in comparison to pure ZnO which increased with the g-C3N4 content (1, 3, 5, and 7 wt.%) in nanocomposites. The photocatalytic activity reached the highest efficiency of 96.8% when the content of g-C3N4 was 7.0 wt.%. Nanocomposite having 7.0 wt.% of g-C3N4 also showed good recyclability with degradation efficiency higher than 90% even in the 4th use. The improvement of photocatalytic activity could be attributed to the adsorption ability and effective separation of electron-hole pairs between g-C3N4 and ZnO. This work implies a simple method to in situ prepare the nanocomposite material of g-C3N4 and semiconductors oxide for photocatalyst applications with high efficiency and good recyclability.

In this report, by a high performance liquid chromatography (HPLC) - high resolution mass spectrometry (HRMS) method, 7 molecular species of phospholipid in the polar lipid class were identified including phosphatidylinositol (PI) and phosphatidylglycerol (PG). The PI 32:0 (16:0/16:0) and PG 34:3 (16:1/18:2) molecular species have the highest content. PI molecular species are mainly formed by saturated fatty acids (16:0, 18:0) and only one polyunsaturated fatty acids C20:4, while PG species are formed by unsaturated fatty acids (16:1, 18:2 and 18:3) and only one saturated fatty acid C16:0.

Hyperphosphatemia is the primary risk factor for vascular calcification, which is closely associated with cardiovascular morbidity and mortality. Recent evidence showed that oxidative stress by high inorganic phosphate (Pi) mediates calcific changes in vascular smooth muscle cells (VSMCs). However, intracellular signaling responsible for Pi-induced oxidative stress remains unclear. Here, we investigated molecular mechanisms of Pi-induced oxidative stress related with intracellular Ca2+ ([Ca2+]i) disturbance, which is critical for calcification of VSMCs. VSMCs isolated from rat thoracic aorta or A7r5 cells were incubated with high Pi-containing medium. Extracellular signal-regulated kinase (ERK) and mammalian target of rapamycin were activated by high Pi that was required for vascular calcification. High Pi upregulated expressions of type III sodium-phosphate cotransporters PiT-1 and -2 and stimulated their trafficking to the plasma membrane. Interestingly, high Pi increased [Ca2+]i exclusively dependent on extracellular Na+ and Ca2+ as well as PiT-1/2 abundance. Furthermore, high-Pi induced plasma membrane depolarization mediated by PiT-1/2. Pretreatment with verapamil, as a voltage-gated Ca2+ channel (VGCC) blocker, inhibited Pi-induced [Ca2+]i elevation, oxidative stress, ERK activation, and osteogenic differentiation. These protective effects were reiterated by extracellular Ca2+-free condition, intracellular Ca2+ chelation, or suppression of oxidative stress. Mitochondrial superoxide scavenger also effectively abrogated ERK activation and osteogenic differentiation of VSMCs by high Pi. Taking all these together, we suggest that high Pi activates depolarization-triggered Ca2+ influx via VGCC, and subsequent [Ca2+]i increase elicits oxidative stress and osteogenic differentiation. PiT-1/2 mediates Pi-induced [Ca2+]i overload and oxidative stress but in turn, PiT-1/2 is upregulated by consequences of these alterations.NEW & NOTEWORTHY The novel findings of this study are type III sodium-phosphate cotransporters PiT-1 and -2-dependent depolarization by high Pi, leading to Ca2+ entry via voltage-gated Ca2+ channels in vascular smooth muscle cells. Cytosolic Ca2+ increase and subsequent oxidative stress are indispensable for osteogenic differentiation and calcification. In addition, plasmalemmal abundance of PiT-1/2 relies on Ca2+ overload and oxidative stress, establishing a positive feedback loop. Identification of mechanistic components of a vicious cycle could provide novel therapeutic strategies against vascular calcification in hyperphosphatemic patients.

Soft tissue augmentation with injectable fillers is increasingly being performed, and providers must be prepared to understand and treat complications. Periorbital skin presents unique challenges due to its thin nature, easily evident pigmentary and textural irregularities, and anatomical considerations including vasculature and subcutaneous layers. The most common complications of infraorbital filler include ecchymosis, malar edema, blue-gray hue, and contour irregularities. Uncommon complications include infection and biofilm formation, skin necrosis, and blindness. This purpose of this review is to highlight the infraorbital anatomy, complications of filler injection in this region, and techniques to avoid these complications.

The aim of the present study is to determine four anionic alkyl sulfate (AS) surfactants with different alkyl chains, namely, C8, C10, C12, and C14, in wastewater by CE with capacitively coupled contactless conductivity detection (CE-C4 D). The conditions effective for the separation of the four AS surfactants were systematically optimized and found to be in a Tris-His (50 mM/20 mM) BGE solution at a pH of 8.95, using a separation voltage of +15 kV, hydrodynamic injection by siphoning using a 20 cm injection height and an injection time of 20 s. The LODs for C8, C10, C12, and C14 were 2.58, 2.30, 2.08, and 3.16 mg/L, respectively. The conditions used to achieve the simultaneous adsorption and preconcentration of the AS surfactants using Al2 O3 beads were pH of 3 and 0.1 mM NaCl. The adsorption efficiencies were found to be 45.6, 50.8, 81.7, and 99.9%, while the desorption efficiencies reached 66.1, 70.4, 83.9, and 100.0% for C8, C10, C12, and C14, respectively. The concentrations of the AS surfactants in wastewater samples were quantified by CE-C4 D after preconcentration by simultaneous adsorption using Al2 O3 beads. The results obtained from the proposed method were consistent with those obtained by HPLC-MS/MS, with a deviation of less than 15%. Our results indicate that the CE-C4 D performed after preconcentration by an adsorption technique using Al2 O3 beads is a new, inexpensive, and suitable method for quantifying AS surfactants in wastewater samples.

Aspergillus fumigatus is a ubiquitous mold that can cause invasive pulmonary infections in immunocompromised patients. Within the lung, A. fumigatus forms biofilms that can enhance resistance to antifungals and immune defenses. Aspergillus biofilm formation requires the production of a cationic matrix exopolysaccharide, galactosaminogalactan (GAG). In this study, recombinant glycoside hydrolases (GH)s that degrade GAG were evaluated as antifungal agents in a mouse model of invasive aspergillosis. Intratracheal GH administration was well tolerated by mice. Pharmacokinetic analysis revealed that although GHs have short half-lives, GH prophylaxis resulted in reduced fungal burden in leukopenic mice and improved survival in neutropenic mice, possibly through augmenting pulmonary neutrophil recruitment. Combining GH prophylaxis with posaconazole treatment resulted in a greater reduction in fungal burden than either agent alone. This study lays the foundation for further exploration of GH therapy in invasive fungal infections. IMPORTANCE The biofilm-forming mold Aspergillus fumigatus is a common causative agent of invasive fungal airway disease in patients with a compromised immune system or chronic airway disease. Treatment of A. fumigatus infection is limited by the few available antifungals to which fungal resistance is becoming increasingly common. The high mortality rate of A. fumigatus-related infection reflects a need for the development of novel therapeutic strategies. The fungal biofilm matrix is in part composed of the adhesive exopolysaccharide galactosaminogalactan, against which antifungals are less effective. Previously, we demonstrated antibiofilm activity with recombinant forms of the glycoside hydrolase enzymes that are involved in galactosaminogalactan biosynthesis. In this study, prophylaxis with glycoside hydrolases alone or in combination with the antifungal posaconazole in a mouse model of experimental aspergillosis improved outcomes. This study offers insight into the therapeutic potential of combining biofilm disruptive agents to leverage the activity of currently available antifungals.

Background and ObjectivesTo expand the clinical knowledge of GPAA1-related glycosylphosphatidylinositol (GPI) deficiency.MethodsAn international case series of 7 patients with biallelic GPAA1 variants were identified. Clinical, biochemical, and neuroimaging data were collected for comparison. Where possible, GPI-anchored proteins were assessed using flow cytometry.ResultsTen novel variants were identified in 7 patients. Flow cytometry samples of 3 available patients confirmed deficiency of several GPI-anchored proteins on leukocytes. Extensive phenotypic information was available for each patient. The majority experienced developmental delay, seizures, and hypotonia. Neuroimaging revealed cerebellar anomalies in the majority of the patients. Alkaline phosphatase was within the normal range in 5 individuals and low in 1 individual, as has been noted in other transamidase defects. We notably describe individuals either less affected or older than the ones published previously.DiscussionClinical features of the cases reported broaden the spectrum of the known phenotype of GPAA1-related GPI deficiency, while outlining the importance of using functional studies such as flow cytometry to aid in variant classification.

Millettia pulchra is traditionally used for treating diseases, including joint pain, fever, anemia, and allergies. It is also a potential resource of natural flavonoid derivatives, which represents major constituents of this plant. This study aimed to isolate the major compounds from M. pulchra radix, develop and validate the HPLC-PDA method to determine their contents, and optimize its extraction. Four major flavonoid derivatives (karanjin, lanceolatin B, 2”,2”-dimethylpyrano-[5″,6″:7,8]-flavone, and pongamol) were isolated using silica gel column chromatography, crystallization techniques in large amounts with high purities (&gt, 95%). A simple, accurate high-performance liquid chromatography–photodiode array (HPLC–PDA) detection method has been developed and validated with significantly statistical impacts according to International Conference on Harmonization (ICH) guidelines. The Response Surface Methodology (RSM), Artificial Neural Network (ANN) models were employed to predictive performance and optimization of the extraction process. The optimized conditions for the extraction of major flavonoids were: extraction time (twice), solvent/material ratio (9.5), and ethanol concentration (72.5%). Our research suggests an effective method, which will be helpful for quality control in the pharmaceutical development of this species.

Vascular calcification (VC) is highly prevalent in patients with atherosclerosis, chronic kidney disease, diabetes mellitus, and hypertension. In blood vessels, VC is associated with major adverse cardiovascular events. Xanthohumol (XN), a main prenylated chalcone found in hops, has antioxidant effects to inhibit VC. This study aimed to investigate whether XN attenuates VC through in vivo study. A rat VC model was established by four weeks oral administration of vitamin D3 plus nicotine in Sprague Dawley (SD) rats. In brief, 30 male SD rats were randomly divided into three groups: control, 25 mg/kg nicotine in 5 mL corn oil and 3 &times, 105 IU/kg vitamin D3 administration (VDN), and combination of VDN with 20 mg/L in 0.1% ethanol of XN (treatment group). Physiological variables such as body and heart weight and drinking consumption were weekly observed, and treatment with XN caused no differences among the groups. In comparison with the control group, calcium content and alkaline phosphatase (ALP) activity were increased in calcified arteries, and XN treatment reduced these levels. Dihydroethidium (DHE) and 2&prime, 7&prime, dichloroflurescin diacetate (DCFH-DA) staining to identify Superoxide and reactive oxygen species generation from aorta tissue showed increased production in VDN group compared with the control and treatment groups. Hematoxylin eosin (HE) and Alizarin Red S staining were determined to show medial vascular thickness and calcification of vessel wall. Administration of VDN resulted in VC, and XN treatment showed improvement in vascular structure. Moreover, overexpression of osteogenic transcription factors bone morphogenetic protein 2 (BMP-2) and runt-related transcription factor 2 (Runx2) were significantly suppressed by XN treatment in VC. Moreover, downregulation of vascular phenotypic markers alpha-smooth muscle actin (&alpha, SMA) and smooth muscle 22 alpha (SM22&alpha, ) were increased by XN treatment in VC. Furthermore, XN treatment in VC upregulated nuclear translocation of nuclear factor-E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) expressions. Otherwise, Kelch-like ECH-associated protein 1 (Keap1) was alleviated by XN treatment in VC. In conclusion, our findings suggested that XN enhances antioxidant capacity to improve VC by regulating the Nrf2/Keap1/HO-1 pathway. Therefore, XN may have potential effects to decrease cardiovascular risk by reducing VC.

Gel polymer electrolyte (GPE) films using polyvinylidene fluoride – hexafluoropropylene (PVdF – HFP) matrix were prepared for aim to achieve good sodium-ion conductivity at room temperature. GPE of PVF-HFP were formed by microporous PVDF-HFP membranes filled and swollen in different liquid electrolytes: GPEs were formed by microporous PVDF-HFP membranes filled and swollen in different liquid electrolytes: NaClO4, NaPF6 and NaTFSI salts dissolved in mixtures of carbonate solvents (propylene carbonate (PC), fluoroethylenecarbonate (FEC)) or ionic liquids (1-ethyl-3-methylimidazolium bis(trifluoromethane-sulfonyl)imide and 1-butyl-3-methyl bis(trifluoromethanesulfonyl)imide). The effect of different electrolytes on the structure and morphology, electrolyte absorption and sodium ionic conductivity of activated membrane were investigated. GPEs showed “sponge-like” structure with high porosity of 80–85% and large pore size. The ionic conductivity of GPEs reached 1–2 mS.cm−1 at room temperature. Sodium intercalation into Na0.44MnO2 electrode was performed in coin-cell type by using GPEs as conduction media. Cycling data of sodium cell using PVdF-HFP/NaClO4 1 M-PC:2%FEC exhibited an excellent stable specific capacity of 100 mAh.g−1 at room temperature.

Desert-like areas located in the eastern portion of the state of Utah (USA) have geographic features that can resemble the surface of the planet Mars, characterized by red-colored hills, soils and sandstones. We examined the bacterial biodiversity of surface soil samples from several sites from the Colorado Plateau Desert in eastern Utah using pyrosequencing of PCR amplified bacterial 16S rRNA genes from total extracted soil DNA. The sample sites cover the Great Basin, Goblin Valley State Park and nearby regions on the Colorado Plateau. We also examined several physicochemical parameters of the soil samples to investigate any possible correlations between bacterial community structure and environmental drivers. The predominant bacterial phyla present in the samples were found to belong to members of the Proteobacteria, Actinobacteria, Bacteroidetes, and Gemmatimonadetes. The most abundant genera in our samples were found to belong to the Cesiribacter, Lysobacter, Adhaeribacter, Microvirga and Pontibacter genera. We found that the relative abundance of Proteobacteria and Gemmatimonadetes were significantly correlated with soil pH and a low concentration of organic matter, suggesting that, in these relatively high-altitude desert soils, these two parameters may be of primary importance to influence bacterial community composition.

Tea and tea products have been known and used for longand became a tradional culture in many countries in theworld, including Vietnam. The quality of tea depends on theconcentrations of typical active substances such as polyphenol groups and their active substances (EGCG, ECG, catechin,…). Therefore, study of analytical methods to determinethese active substances is necessary to evaluate the qualityof tea and tea products. In this study, UV-Vis method was selected to determine total polyphenol and HPLC method wasselected to determine the main active ingredient of polyphenolgroup EGCG. The limit of detections are 0.12 mg/g and 0.064mg/g for total polyphenol and EGCG, respectively. Thesemethods were applied to determine the concentrations of totalphenol and EGCG in 10 samples of tea and tea products. Results show that the concentrations of total phenol and EGCGvary significantly among samples, but there is a certain relationship between the concentration of total phenol and EGCGin each sample.

The oxidative dehydrogenation of allyl alcohol to acrolein over bulk alloyed and supported silver catalysts has been studied, with the aim of designing optimal Ag-based catalysts for this specific reaction. All of the tested catalysts were highly active and selective to acrolein at temperatures ranging between 340 and 380 °C. The results show that the alloying of Ag with Au or Cu leads to alloys with upper surface compositions comprising approximately between 35 and 65% silver. Although these elements improve the activity of Ag catalysts, they decrease their selectivity, leading to lower acrolein yields. Various different Ag catalyst supports were prepared (LaPO4, Carbon nanotubes CNT, TiO2, Al2O3 and SiO2) using the impregnation method, and subsequently tested. This study reveals that the catalyst with the best performance is Ag/SiO2 with a yield in acrolein of 87.2% at 360 °C. The results obtained with the tested catalysts are discussed, and the reasons of performance improvement or degradation caused by the presence of Au or Cu on the active silver sites, or by the presence of the supports, are raised.

© The Author(s). 2017. Background: Antimicrobial resistance has emerged as a major concern in developing countries. The present study sought to define the pattern of antimicrobial resistance in ICU patients with ventilator-associated pneumonia. Methods: Between November 2014 and September 2015, we enrolled 220 patients (average age ~ 71 yr) who were admitted to ICU in a major tertiary hospital in Ho Chi Minh City, Vietnam. Data concerning demographic characteristics and clinical history were collected from each patient. The Bauer-Kirby disk diffusion method was used to detect the antimicrobial susceptibility. Results: Antimicrobial resistance was commonly found in ceftriaxone (88%), ceftazidime (80%), ciprofloxacin (77%), cefepime (75%), levofloxacin (72%). Overall, the rate of antimicrobial resistance to any drug was 93% (n = 153/164), with the majority (87%) being resistant to at least 2 drugs. The three commonly isolated microorganisms were Acinetobacter (n = 75), Klebsiella (n = 39), and Pseudomonas aeruginosa (n = 29). Acinetobacter baumannii were virtually resistant to ceftazidime, ceftriaxone, piperacilin, imipenem, meropenem, ertapenem, ciprofloxacin and levofloxacin. High rates ( > 70%) of ceftriaxone and ceftazidime-resistant Klebsiella were also observed. Conclusion: These data indicated that critically ill patients on ventilator in Vietnam were at disturbingly high risk of antimicrobial resistance. The data also imply that these Acinetobacter, Klebsiella, and Pseudomonas aeruginosa and multidrug resistance pose serious therapeutic problems in ICU patients. A concerted and systematic effort is required to rapidly identify high risk patients and to reduce the burden of antimicrobial resistance in developing countries.

There are over 150 known human proteins which are tethered to the cell surface via glycosylphosphatidylinositol (GPI) anchors. These proteins play a variety of important roles in development, and particularly in neurogenesis. Not surprisingly, mutations in the GPI anchor biosynthesis and remodeling pathway cause a number of developmental disorders. This group of conditions has been termed inherited GPI deficiencies (IGDs), a subgroup of congenital disorders of glycosylation; they present with variable phenotypes, often including seizures, hypotonia and intellectual disability. Here, we report two siblings with compound heterozygous variants in the gene phosphatidylinositol glycan anchor biosynthesis, class P (PIGP) (NM_153681.2: c.74T > C;p.Met25Thr and c.456delA;p.Glu153AsnFs*34). PIGP encodes a subunit of the enzyme that catalyzes the first step of GPI anchor biosynthesis. Both children presented with early-onset refractory seizures, hypotonia, and profound global developmental delay, reminiscent of other IGD phenotypes. Functional studies with patient cells showed reduced PIGP mRNA levels, and an associated reduction of GPI-anchored cell surface proteins, which was rescued by exogenous expression of wild-type PIGP. This work associates mutations in the PIGP gene with a novel autosomal recessive IGD, and expands our knowledge of the role of PIG genes in human development.

NdPO4 has been studied as a model rare earth phosphate catalyst for the dehydration of 2,3-butanediol (2,3-BDO) to butadiene (BD), in order to understand the influence of structure and morphology o...

It is estimated that 0.5% of all mammalian proteins have a glycosylphosphatidylinositol (GPI)-anchor. GPI-anchored proteins (GPI-APs) play key roles, particularly in embryogenesis, neurogenesis, immune response and signal transduction. Due to their involvement in many pathways and developmental events, defects in the genes involved in their synthesis and processing can result in a variety of genetic disorders for which affected individuals display a wide spectrum of features. We compiled the clinical characteristics of 202 individuals with mutations in the GPI biosynthesis and processing pathway through a review of the literature. This review has allowed us to compare the characteristics and the severity of the phenotypes associated with different genes as well as highlight features that are prominent for each. Certain combinations, such as seizures with aplastic/hypoplastic nails or abnormal alkaline phosphatase levels suggest an inherited GPI deficiency, and our review of all clinical findings may orient the management of inherited GPI deficiencies.

La, Nd, Sm and Gd rare earth orthophosphates with a Rhabdophane-type structure have been synthesized and tested as catalysts for the dehydration of ethanol and 1-butanol. These solids appear to be very active and selective. Contrary to a previously studied LaPO4 series of catalysts, whose properties were shown to depend on their P/La surface ratio, this series of rare earth phosphates exhibits catalytic properties depending mainly on the nature of the rare earth elements themselves. The most efficient phosphates presented moderately strong Bronsted and Lewis acid sites and only small quantities of weak basic sites. These properties lead to an exceptionally high selectivity to alkenes for both studied dehydration reactions. Although a good correlation was observed between the Lewis acidity and the rate of ethanol conversion, a possible involvement of Bronsted acid sites, the strength of which increases with that of the Lewis sites, cannot be totally excluded. The results of the catalysts characterization are interpreted in terms of the possible nature of the different sites.

Methane hazard has always been considered for underground coal mining as it can lead to methane explosion. In Quang Ninh province, several coal mines such as Mạo Khe coal mine, Khe Cham coal mine, especially Duong Huy mine that have high methane content. Experimental data to examine contents of methane bearing coal seams at different depths are not similar in Duong coal mine. In order to ensure safety, this report has been undertaken to determine a pattern of changing methane contents of coal seams at different exploitation depths in Duong Huy underground coal mine.

Four new lignans, a furofuran lignan medioresinol B (10) and three tetrahydrofuran lignans kobusinol C (16), 7'-methoxy magnostellin A (21), and mangnostellin D (23), along with 19 known lignans, were isolated from the flower buds of Magnolia biondii PAMP. The structures of the isolates were elucidated using spectroscopic analysis, mainly one- and two-dimensional NMR, high resolution-MS, and circular dichroism techniques as well as Mosher's esterification method. The anti-allergic effects of the isolated compounds were evaluated by analyzing the inhibition of interleukin-2 (IL-2) expression in Jurkat T-cells. Compounds 11-14 reduced IL-2 expression in a dose-dependent manner.

Many studies have comprehensively examined the venom ofOphiophagus hannahsnake. Its venom comprises different compounds exhibiting a wide range of pharmacological activities. In this investigation, four peptide fractions (PFs), ranging from 3 kDa to 10 kDa, isolated from the Vietnamese snake venom ofO. hannahwere separated by HPLC and investigated for their inhibitory activity on adipogenesis in 3T3-L1 adipocytes. The most effective PF was then further purified, generating two peptides, pOh1 and pOh2. Upon investigation of these two peptides on 3T3-L1 adipocytes, it was revealed that, at 10 μg/mL, pOh2 was able to inhibit the lipid accumulation in 3T3-L1 adipocytes by up to 56%, without affecting cell viability. Furthermore, the pOh2 downregulated the gene expression of important transcription factors C/EBP-αand PPAR-γ. In addition, aP2 and GPDH adipocyte-specific markers were also significantly reduced compared to untreated differentiated cells. Taken together, pOh2 inhibited the expression of key transcription factors C/EBP-αand PPAR-γand their target genes, aP2 and GPDH, thereby blocking the adipocyte differentiation. In conclusion, this novel class of peptide might have potential for in vivo antiobesity effects.

International audience; Insulin secretion by pancreatic β-cells in response to glucose or other secretagogues is tightly coupled to membrane potential. Various studies have highlighted the prospect of enhancing insulin secretion in a glucose-dependent manner by blocking voltage-gated potassium channels (K v) and calcium-activated potassium channels (K Ca). Such strategy is expected to present a lower risk for hypoglycemic events compared to K ATP channel blockers. Our group recently reported the discovery of a new insulinotropic agent, cardiotoxin-I (CTX-I), from the Naja kaouthia snake venom. In the present study, we report the design and synthesis of [Lys 52 ]CTX-I 41−60 via structure-guided modification, a truncated, equipotent analogue of CTX-I, and demonstrate, using various pharmacological inhibitors, that this derivative probably exerts its action through K v channels. This new analogue could represent a useful pharmacological tool to study β-cell physiology or even open a new therapeutic avenue for the treatment of type 2 diabetes.

One diazonium (perfluoroalkyl) benzenesulfonimide monomer, perfluoro-3, 6-dioxa-4-methyl-7-octene benzenesulfonyl imide, has been synthesized from Nafion monomer for the first time. With trifluorovinyl ether and diazonium precursors, the partially-fluorinated diazonium PFSI monomer can be polymerized and will provide chemically bonding with carbon electrode in proton exchange membrane fuel cells. A systematic study of the synthesis and characterization of this diazonium PFSI monomer has been conducted by varying reaction conditions. The optimized synthesis method has been established in the lab.

Two new tetraoxygenated xanthones, 6- O -demethyloliverixanthone and schomburgxanthone, together with cowanin, cowanol, fuscaxanthones A and B, 3-isomangostin hydrate, and 1,7-dihydroxyxanthone, were isolated from the bark of Garcinia schomburgkiana . Their structures were elucidated using 1-D and 2-D NMR techniques as well as chemical methods. Five of the isolated compounds were tested regarding their cytotoxicity against HeLa cells and the results showed that fuscaxanthone B and cowanin possessed remarkable activity with IC 50 values of 2.4 ± 0.2 and 2.7 ± 0.3 μg/ml, respectively, using an MTT assay.

Four complete beef-production systems consisting each of two stages were compared. The systems were formed by combining two diets for the cow-calf herd with finishing heifers stage – St (Standard) and O3 (maximising omega-3 fatty acids (FAs) using wrapped grass silage) – with four diets for the bull-fattening herd stage — SM (silage maize starch), SML (silage maize starch plus linseed, rich in omega-3 FAs), FC (fibre-based concentrate), and SCL (starch-based concentrate plus linseed): St-SM, O3-SML, St-FC and O3-SCL. Life Cycle Assessments applied to these systems (from cradle to farm gate for a one-year period) estimated that their environmental impacts, per kg of carcass mass, ranged from 27.0 to 27.9 kg CO 2 equivalents (eq), 64.8–73.4 MJ, 94–98 g PO 4 3 − eq, 168–173 g SO 2 eq, 47–48 m 2 year for climate change (CC, not including effect of land use and land-use change, LULUC), cumulative energy demand (CED), eutrophication potential, acidification potential and land occupation, respectively. Consideration of LULUC decreased CC from 8 to 10%. Minor impact differences between these systems were observed, except for CED of St-FC, mainly because more energy was needed to dehydrate beet pulp and lucerne. CC of O3-SCL was 3% lower than CC of St-SM. Most of the environmental impacts of beef-production systems originated from the cow-calf herd with finishing heifers (73–97%), which indicates that research on the reduction of environmental impacts of this type of beef-production system should focus on this herd. For the cow-calf herd with finishing heifers, comparison of several allocation methods revealed that allocation method strongly affected the impacts per kg of carcass mass of the breeding bull and finished cull cows and, to a much lesser extent, those of fattened bulls and finished heifers. Consideration of both products (several animal types) and the ecosystem services supplied by these systems seems a promising perspective. This concept needs to be discussed and developed as an approach to consider the multi-functionality of farming systems.

XPS:RMN+ECI2D+LMA:CLO:PAF:JMI; International audience; Four preparation methods have been developed for the synthesis of lanthanum orthophosphate catalysts, which are shown to be active and selective catalysts for the dehydration of light alcohols (C-2 to C-4). Their crystallinity, surface area, surface composition and acid-base properties appear to differ according to the preparation method used. The most important parameter influencing their catalytic properties appears to be their surface composition since this has a direct influence on their surface acidity. All of the solids presented weak and moderately strong Bronsted and Lewis acid sites, and only weak basic sites in low quantities. These acid-base properties resulted in the prevalence of an E-1-type mechanism for 1-butanol dehydration. Bronsted acid sites, which appear as the most efficient sites, are associated with the presence of an excess of phosphorus at their surface and from spectroscopic data this is attributed to (H2PO4)(-3+n) (n=1 or 2) species from spectroscopic data. The prevalence of these species and their optimum surface distribution make them extraordinarily efficient and ultra-selective for the dehydration of several alcohols. Published by Elsevier B.V.

Inorganic phosphate (Pi) plays an important role in cell signaling and energy metabolism. In insulin-releasing cells, Pitransport into mitochondria is essential for the generation of ATP, a signaling factor in metabolism-secretion coupling. Elevated Piconcentrations, however, can have toxic effects in various cell types. The underlying molecular mechanisms are poorly understood. Here, we have investigated the effect of Pion secretory function and apoptosis in INS-1E clonal β-cells and rat pancreatic islets. Elevated extracellular Pi(1∼5 mM) increased the mitochondrial membrane potential (ΔΨm), superoxide generation, caspase activation, and cell death. Depolarization of the ΔΨmabolished Pi-induced superoxide generation. Butylmalonate, a nonselective blocker of mitochondrial phosphate transporters, prevented ΔΨmhyperpolarization, superoxide generation, and cytotoxicity caused by Pi. High Pialso promoted the opening of the mitochondrial permeability transition (PT) pore, leading to apoptosis, which was also prevented by butylmalonate. The mitochondrial antioxidants mitoTEMPO or MnTBAP prevented Pi-triggered PT pore opening and cytotoxicity. Elevated extracellular Pidiminished ATP synthesis, cytosolic Ca2+oscillations, and insulin content and secretion in INS-1E cells as well as in dispersed islet cells. These parameters were restored following preincubation with mitochondrial antioxidants. This treatment also prevented high-Pi-induced phosphorylation of ER stress proteins. We propose that elevated extracellular Picauses mitochondrial oxidative stress linked to mitochondrial hyperpolarization. Such stress results in reduced insulin content and defective insulin secretion and cytotoxicity. Our data explain the decreased insulin content and secretion observed under hyperphosphatemic states.

Ten sesquiterpenoids (1-10) and six phenylpropanoid derivatives (11-16) were isolated from the flower buds of Magnolia biondii Pamp. Their structures were determined by 1D- and 2D-NMR spectroscopy, mass spectrometry, and optical rotation. To evaluate their anti-allergic properties, the inhibitory effect of each isolate (1-16) on interleukin-2 (IL-2) gene expression was examined in Jurkat T cells. Among the isolated compounds, three sesquiterpenoids (2, 5, and 7) and two monoterpenoids (12 and 13) strongly inhibited IL-2 production in Jurkat T cells. Four compounds, (-)-parthenolide (2), eudesm-4(15)-eno-1β,6α-diol (5), biondinin D (12), and tiliroside (16), showed cytotoxicity against Jurkat T cells. In addition, (-)-parthenolide (2) exhibited cytotoxicity against the human cervical cancer HeLa cell line, the human breast cancer MCF-7, and human promyelocytic leukemia HL-60 cell lines.

The C-terminal fragment of the c-Met receptor tyrosine kinase is present in the nuclei of some cells irrespective of ligand stimulation, but the responsible nuclear localization signal (NLS) has not been previously reported. Here, we report that two histidine residues separated by a 10-amino-acid spacer (H1068-H1079) located in the juxtamembrane region of c-Met function as a putative novel NLS. Deletion of these sequences significantly abolished the nuclear translocation of c-Met, as did substitution of the histidines with alanines. This substitution also decreased the association of c-Met fragment with importin β. The putative NLS of c-Met is unique in that it relies on histidines, whose positive charge changes depending on pH, rather than the lysines or arginines, commonly found in classical bipartite NLSs, suggesting the possible 'pH-dependency' of this NLS. Indeed, decreasing the cytosolic pH either with nigericin, an Na(+)/H(+) exchanger or pH 6.5 KRB buffer significantly increased the level of nuclear c-Met and the interaction of the c-Met fragment with importin β, indicating that low pH itself enhanced nuclear translocation. Consistent with this, nigericin treatment also increased the nuclear level of endogenous c-Met in HeLa cells. The putative aberrant bipartite NLS of c-Met seems to be the first example of what we call a 'pH-dependent' NLS.

Three trypsin inhibitors (TIs), from the seeds of the squash Momordica cochinchinensis (MCo), have been isolated and purified using gel filtration, ion exchange chromatography, and reverse-phase HPLC. Their sequences could be determined only after proteolytic cleavages. In the case of MCoTI-I and -II, it was shown that their polypeptide backbones are cyclic, a structure that has never been described in squash TIs. They contain 34 amino acid residues with 3 disulfide bridges and measured molecular masses of 3453.0 and 3480.7, respectively. They are the largest known macrocyclic peptides containing disulfide bridges. Their sequences show strong homology to other squash TIs, suggesting a similar three-dimensional structure and an analogous mechanism of action. A model of MCoTI-II was constructed by analogy to the crystal structure of the complex between bovine trypsin and CMTI-I, indicating that the linker connecting the two termini is flexible and does not impose significant geometrical constraints. This flexibility allows an Asp-Gly peptide bond rearrangement to occur in this region, giving rise to two isoforms of MCoTI-II. Although the importance of cyclization is not clear, it might confer increased stability and resistance to proteolysis. A minor species, MCoTI-III, was also characterized as containing 30 amino acid residues with a molecular mass of 3379.6. This component possesses a linear backbone with a blocked N-terminus. MCoTIs represent interesting candidates for drug design, either by changing their specificity of inhibition or by using their structure as natural scaffolds bearing new binding activities.

International audience; The urotensinergic system plays central roles in the physiological regulation of major mammalian organ systems, including the cardiovascular system. As a matter of fact, this system has been linked to numerous pathophysiological states including atherosclerosis, heart failure, hypertension, diabetes as well as psychological, and neurological disorders. The delineation of the (patho)physiological roles of the urotensinergic system has been hampered by the absence of potent and selective antagonists for the urotensin II-receptor (UT). Thus, a more precise definition of the molecular functioning of the urotensinergic system, in normal conditions as well as in a pathological state is still critically needed. The recent discovery of nuclear UT within cardiomyocytes has highlighted the cellular complexity of this system and suggested that UT-associated biological responses are not only initiated at the cell surface but may result from the integration of extracellular and intracellular signaling pathways. Thus, such nuclear-localized receptors, regulating distinct signaling pathways, may represent new therapeutic targets. With the recent observation that urotensin II (UII) and urotensin II-related peptide (URP) exert different biological effects and the postulate that they could also have distinct pathophysiological roles in hypertension, it appears crucial to reassess the recognition process involving UII and URP with UT, and to push forward the development of new analogs of the UT system aimed at discriminating UII- and URP-mediated biological activities. The recent development of such compounds, i.e. urocontrin A and rUII(1-7), is certainly useful to decipher the specific roles of UII and URP in vitro and in vivo. Altogether, these studies, which provide important information regarding the pharmacology of the urotensinergic system and the conformational requirements for binding and activation, will ultimately lead to the development of potent and selective drugs.

The effect of Taylor vortices flow of Couette-Taylor (CT) crystallizer on the nucleation and reconstruction of L-glutamic acid was firstly investigated in cooling crystallization. Generally, L-glutamic acid has two kinds of polymorphic crystal including metastable α-form and stable β-form, where the α-form is initially crystallized and then slowly transformed into the β-form, which is called the phase transformation. The present study explored that there was a significant difference between CT and the conventional ST crystallizer as regards the nucleation and reconstruction of L-glutamic acid. Here, the phase transformation determined by the nucleation and reconstruction was 40h in ST crystallizer, yet it was only 20h as using the CT crystallizer, implying that the nucleation and reconstruction of L-glutamic acid was facilitated 2.0 times as using the Taylor vortices flow. The advantage of Taylor vortices flow in CT crystallizer over the random fluid motion in ST crystallizer with regard to the nucleation and reconstruction was explained in terms of the high energy dissipation of Taylor vortices flow.

The transcription factor LEAFY COTYLEDON1 (LEC1) controls aspects of early embryogenesis and seed maturation in Arabidopsis thaliana. To identify components of the LEC1 regulon, transgenic plants were derived in which LEC1 expression was inducible by dexamethasone treatment. The cotyledon-like leaves and swollen root tips developed by these plants contained seed-storage compounds and resemble the phenotypes produced by increased auxin levels. In agreement with this, LEC1 was found to mediate up-regulation of the auxin synthesis gene YUCCA10. Auxin accumulated primarily in the elongation zone at the root-hypocotyl junction (collet). This accumulation correlates with hypocotyl growth, which is either inhibited in LEC1-induced embryonic seedlings or stimulated in the LEC1-induced long-hypocotyl phenotype, therefore resembling etiolated seedlings. Chromatin immunoprecipitation analysis revealed a number of phytohormone- and elongation-related genes among the putative LEC1 target genes. LEC1 appears to be an integrator of various regulatory events, involving the transcription factor itself as well as light and hormone signalling, especially during somatic and early zygotic embryogenesis. Furthermore, the data suggest non-embryonic functions for LEC1 during post-germinative etiolation.

There has been controversy over whether a short period of cardiopulmonary resuscitation (CPR) prior to defibrillation improves survival in patients who experienced a sudden cardiac arrest. However, there have been no reports about whether CPR restores the myocardial energy source during prolonged ventricular fibrillation (VF). The aim of this study is to investigate the effect of CPR in restoring myocardial high energy phosphates during prolonged VF.Seventy-two adult male Sprague-Dawley rats were used in this study. Baseline adenosine triphosphate (ATP) and adenosine diphosphate (ADP) prior to induction of VF were measured in nine rats, the No-VF group. Sixty-three rats were subjected to 4 min of untreated VF. Animals were then randomized into two groups: No-CPR (n=37) and CPR (n=26). In the No-CPR group, ATPs and ADPs were measured at 4 min (No-CPR4), 6 min (No-CPR6), 8 min (No-CPR8) or 10 min (No-CPR10) after the induction of VF. The CPR group received 2 min (CPR2), 4 min (CPR4) or 6 min (CPR6) of mechanical chest compressions before ATP was measured. Myocardial ATP (nmol/mg protein) was decreased as VF duration was prolonged (No-VF: 5.49±1.71, No-CPR4: 4.27±1.58, No-CPR6: 4.13±1.31, No-CPR8: 3.77±1.42, No-CPR10: 3.52±0.90, p0.05 between each of No-CPRs vs. No-VF). Two minutes of CPR restored myocardial ATP to the level of No-VF group (5.27±1.67 nmol/mg protein in CPR2, p0.05 vs. No-VF group). However, myocardial ATP (nmol/mg protein) decreased if the duration of CPR was longer than 2 min (CPR4: 3.77±1.05, CPR6: 3.49±1.08, p0.05 between CPR4 and CPR6 vs. No-VF).CPR for 2 min helps to maintain myocardial ATP after prolonged VF.

International audience; Urotensin II (UII) and its receptor UT, are widely expressed in the cardiovascular and central nervous system, where they exert regulatory actions under both physiological and pathological conditions. Our study, aimed at investigating the presence of functional nuclear UT in various rat and monkey tissues as well as in human cell lines, demonstrated for the first time by Western blot analysis and confocal immunofluorescence a tissue-specific nuclear expression of this receptor (heart and central nervous system). This nuclear UT was further characterized pharmacologically through radioligand binding studies using specific ligands of the urotensinergic system, as well as somatostatin. In 2D-gel experiments, we observed the presence of different post-translational modifications between membrane and nuclear UT receptors in brain extracts. Transcription initiation assays showed de novo RNA synthesis caused by UII and Urotensin-related peptide (URP) which were inhibited by an UT antagonist urantide. In hypoxic/ischemic conditions, UT receptors were differentially modulated in regard to subcellular localization. Thus, the unique regiospecificity of the nuclear UT receptor along with its particular modulation under hypoxic conditions could indicate a specific and complementary physiological role that could be correlated with pro-angiogenic and/or neuromodulatory actions of UII, both in the cardiovascular and central nervous system.

Publisher Summary Cocoa butter (CB) is an essential ingredient in chocolate as it forms the continuous phase of chocolate. It is responsible for the gloss, texture, and typical melting behavior of chocolate. Although cocoa butter is the ideal ingredient, the varying supply and increasing price depending on fluctuating cocoa bean prices forced manufacturers to seek alternatives. Cocoa butter can be replaced by other vegetable fats, collected under the name Cocoa Butter Alternatives (CBAs). The CBAs are divided into three main categories according to their functionality and similarity to cocoa butter: the Cocoa Butter Equivalents (CBEs), the Cocoa Butter Replacers (CBRs), and the Cocoa Butter Substitutes (CBSs). A CBE should allow processing of chocolate products in an identical manner to that of cocoa butter-based products. The specific fatty acid profile and their distribution on the glycerol backbone in natural oils and fats affect their functional properties like crystallization and melting behavior. Therefore, the use of only one technique is not sufficient and several techniques need to be combined together to produce a cocoa butter alternative fat. Interesterification is an important modification technique resulting in the redistribution of the fatty acids along the glycerol backbone, which results in a change of the physicochemical properties of the fat.

+MAO; MoVTe(Sb)NbO catalysts have been prepared using the slurry method and tested as catalysts for the oxidative dehydrogenation of ethane. The influence of both the addition of silica to the starting slurry during the preparation and of a final heat treatment under nitrogen at 873 K after the dissolution of the M2 phase formed concomitantly to the active M1 active phase has been studied. While the parameters change the texture of the catalysts without modification of the catalytic sites, they have opposite effects on catalytic properties. The addition of silica increases the conversion without modifying the selectivity to ethylene and the final heat treatment decreases the conversion but increases the selectivity. These effects have been explained respectively by a decrease of the agglomeration and sintering of the M1 phase crystallites and a decrease of the number of small pores, where total oxidation takes place. These effects could be leveraged to optimize the preparation protocol and obtain more efficient catalysts for the oxidative dehydrogenation of ethane. (C) 2012 Elsevier B.V. All rights reserved.

A normal-phase high-performance liquid chromatography-evaporative light-scattering detector method employing dichloromethane, methanol, and acetic acid/triethylamine buffer as the mobile phase was developed for analysis of polar lipids (PLs). This method was applicable for analysis of PLs from both dairy materials and soy lecithin. All of the PLs of interest such as glycolipids, phospholipids, and sphingomyelin were well separated with a total run time of 22.5 min and without necessitating the removal of neutral lipids beforehand. Peak retention times were stable, and the method was reproducible. In this study, a modified method of using solvents for extraction of PLs from dairy matrices was also investigated. The modified method offered higher extraction efficiency, consumed less time, and in some cases saved solvent use.

We investigated to establish CD40-activated B cells (CD40-B cells) as alternative antigen-presenting cells (APCs) for the induction of myeloma-specific cytotoxic T lymphocytes (CTLs). To generate CD40-B cells, peripheral blood mononuclear cells were co-cultured with CD40L-transfected J558 cells in the presence of IL-4, insulin, transferrin, and cyclosporine for 14 days, and pulsed with myeloma lysates. The CD40-B cells consistently expressed high levels of CD80, CD86, CD54, CCR7, and HLA-DR. The CD40-B cells produced IL-12, IFN-gamma, and IL-6 during the culture period, but not IL-10. In addition, the CD40-B cells showed potent allogeneic T-cell stimulatory capacities that depended on the dose ratio and had the potential to polarize naïve T cells into Th1 subsets. The CD40-B cells loaded with tumor lysates induced strong target-specific CTLs, based on large numbers of IFN-gamma secreting cells and higher cytotoxic activity against target cells compared to the CD40-B cells without the tumor lysates. These results suggest that CD40-B cells loaded with myeloma lysates might provide alternative APCs for cellular immunotherapy in patients with myeloma.

Squash trypsin inhibitors are small disulfide-rich proteins (28–30 residues, 6 cysteines). Their 3D structures revealed that one disulfide crosses the macrocycle formed by the other two disulfide bridges and the interconnecting backbone, hence the term “knottins”. This particular topology has been found in a number of cystine-rich peptides with diverse biological activities isolated from plants or animals. Several of these peptides, such as kalata Bl, circulin A and B and Cycloviolacin 01, display macrocyclic structures. These cyclic knottins were grouped in two subfamilies following sequence comparison [1]. To date, all the reported squash trypsin inhibitors have a linear structure. We recently isolated several cyclic squash trypsin inhibitors from Momordica cochinchinensis seeds [2]. MCoTI-II is the most abundant and its primary structure has been identified as cyclo-(S1 G S D G G V C P K10 I L K K C R R D S D20 C P G A C I C R G N30 G Y C G). Here we report the NMR and molecular modeling studies of this peptide.

The solvothermal reactions of 1,4-bezenedicarboxylic acid (H 2 BDC) or azobenzene-4,4′-dicarboxylic acid (H 2 ABD) with zinc ions/clusters lead to the formation of four crystalline materials. All of these compounds were characterized by x-ray diffraction, optical microscopy, thermo-gravimetric analysis and nitrogen adsorption. Block-shaped crystals (BZ1) with various shapes and sizes were obtained at H 2 BDC:Zn mole ratio of 1:1 and H 2 BDC concentration of 0.1 M. At more dilute H 2 BDC concentration of 0.01 M and H 2 BDC:Zn mole ratio of 1 : 4, the reaction product was cubic crystals (BZ2) with a size of 250 μm. In the H 2 ABD system, flat-plate-like crystals (AZ1) were obtained at H 2 ABD:Zn mole ratio of 1 : 1 and H 2 ABD concentration of 0.01 M. Meanwhile, thick-block-like crystals (AZ2) were formed at the same H 2 ABD:Zn mole ratio but at 0.004 M H 2 ABD. The Langmuir surface area (S Lang ) of the materials was remarkable, enhanced by diluting the reaction solution. For the compounds synthesized in N , N′-dimethylformamide (DMF), S Lang increased from 304.6 m 2 g −1 for BZ1 to 2631 m 2 g −1 for BZ2 and from 475.8 m 2 g −1 for AZ1 to 3428 m 2 g −1 for AZ2. Meanwhile, BZ2 synthesized in N , N′-diethylformamide (BZ2/DEF) got the highest S Lang of 4330 m 2 g −1. Both AZ2 and BZ2 materials were stable up to 400 °C.