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1.
Biochem Biophys Res Commun ; 548: 47-52, 2021 04 09.
Artigo em Inglês | MEDLINE | ID: mdl-33631673

RESUMO

The mechanism for protein stabilization or destabilization has long been an open quest. In the present study, we have studied the interactions between amino acids and guanidinium (Gdm+)/ammonium (NH4+) ions by using low field nuclear magnetic resonance (LF-NMR), where Gdm+ and NH4+ are denaturant and stabilizer for proteins, respectively. It shows that Gdm+ favors to bind to the thiol group or the hydroxyl group on the side chain but weakly interacts with the α-carboxyl group. In contrast, NH4+ prefers to bind to the α-carboxyl group but slightly interacts with the thiol group or the hydroxyl group on the side chain of amino acids. 1HNMR reveals the hydrogen bonding between NH4+ and the α-carboxyl group, which is not involved in the interactions between Gdm+ and cysteine. Our study demonstrates that the strong interactions between the denaturant and the sulfur atom or the disulfide bond promote the direct binding of the denaturant toward proteins, leading to the destabilization.


Assuntos
Aminoácidos/química , Cloreto de Amônio/química , Cátions , Guanidina/química , Hidrogênio , Estabilidade Proteica , Espectroscopia de Prótons por Ressonância Magnética , Soluções
2.
Chem Biodivers ; 17(9): e2000258, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32638471

RESUMO

In the present study, coumarin-bearing three pyridinium and three tetra-alkyl ammonium salts were synthesized. The compounds were fully characterized by 1 H- and 13 C-NMR, LC/MS and IR spectroscopic methods and elemental analyses. The cytotoxic properties of all compounds were tested against human liver cancer (HepG2), human colorectal cancer (Caco-2) and non-cancer mouse fibroblast (L-929) cell lines. Some compounds performed comparable cytotoxicity with standard drug cisplatin. Antibacterial properties of the compounds were tested against Gram-negative Escherichia coli and Gram-positive Bacillus subtilis bacteria, but the compounds did not have any antibacterial effect against both bacteria. Enzyme inhibitory properties of all compounds were tested on the activities of human carbonic anhydrase I and II, and xanthine oxidase. All compounds inhibited both enzymes more effectively than standard drugs, acetazolamide and allopurinol, respectively. The biological evaluation results showed that ionic and water soluble coumarin derivatives are promising structures for further investigations especially on enzyme inhibition field.


Assuntos
Cloreto de Amônio/farmacologia , Antibacterianos/farmacologia , Antineoplásicos/farmacologia , Cumarínicos/farmacologia , Inibidores Enzimáticos/farmacologia , Cloreto de Amônio/síntese química , Cloreto de Amônio/química , Antibacterianos/síntese química , Antibacterianos/química , Antineoplásicos/síntese química , Antineoplásicos/química , Bacillus subtilis/efeitos dos fármacos , Anidrases Carbônicas/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cumarínicos/síntese química , Cumarínicos/química , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Escherichia coli/efeitos dos fármacos , Humanos , Testes de Sensibilidade Microbiana , Estrutura Molecular , Solubilidade , Relação Estrutura-Atividade , Água/química , Xantina Oxidase/antagonistas & inibidores , Xantina Oxidase/metabolismo
3.
Chemosphere ; 238: 124700, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31524602

RESUMO

An eight-year field trial was conducted to investigate the effects of four different N fertilization treatments of urea (CO(NH2)2, the control), ammonium sulfate ((NH4)2SO4), ammonium chloride (NH4Cl), and ammonium hydrogen phosphate [(NH4)2HPO4]) on cadmium (Cd) phytotoxicity in rice and soil microbial communities in a Cd-contaminated paddy of southern China. The results demonstrate that the different N treatments exerted different effects: the application of (NH4)2HPO4 and (NH4)2SO4 significantly increased rice grain yield and decreased soil-extractable Cd content when compared with those of the control, while NH4Cl had a converse effect. Expression of genes related to Cd uptake (IRT and NRAPM genes) and transport (HMA genes) by roots may be responsible for Cd phytotoxicity in rice grown in the different N fertilization treatments. Our results further demonstrate that N fertilization had stronger effects on soil bacterial communities than fungal communities. The bacterial and fungal keystone species were identified by phylogenetic molecular ecological network (pMEN) analysis and mainly fell into the categories of Gammaproteobacteria, Acidobacteria and Actinobacteria for the bacterial species and Ascomycota for the fungal species; all of these keystone species were highly enriched in the (NH4)2HPO4 treatment. Soil pH and soil available-Cd content emerged as the major determinants of microbial network connectors. These results could provide effective fertilizing strategies for alleviating Cd phytotoxicity in rice and enhance the understanding of its underlying microbial mechanisms.


Assuntos
Ascomicetos/isolamento & purificação , Bactérias/isolamento & purificação , Cádmio/análise , Cádmio/toxicidade , Oryza/química , Poluentes do Solo/toxicidade , Acidobacteria/isolamento & purificação , Actinobacteria/isolamento & purificação , Cloreto de Amônio/química , Sulfato de Amônio/química , Ascomicetos/metabolismo , Bactérias/metabolismo , China , Grão Comestível/química , Poluição Ambiental/análise , Fertilizantes/análise , Gammaproteobacteria/isolamento & purificação , Microbiota , Oryza/microbiologia , Fosfatos/química , Filogenia , Raízes de Plantas/metabolismo , Raízes de Plantas/microbiologia , Solo/química , Microbiologia do Solo , Poluentes do Solo/análise , Ureia/química
4.
J Pharm Biomed Anal ; 180: 113034, 2020 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-31838281

RESUMO

Official method in Ph. Eur. for evaluation of timolol enantiomeric purity is normal-phase high performance liquid chromatography (NP-HPLC) method. Compared to other HPLC modes, NP is depicted as quite expensive with high consumption of organic solvents which leads to chronic exposure of analysts to toxic and carcinogenic effects. In order to overcome above-mentioned drawbacks, the aim of this study was to develop new method with better eco-friendly features. This was enabled by using protein type Chiral Stationary Phase (CSP) in reversed-phase mode that required up to 10 % (v/v) of organic solvent. Therefore, an enantioselective HPLC method was developed and validated for quantification of (S)-timolol and its chiral impurity, (R)-isomer. Optimized separation conditions on ovomucoid column were set using Analytical Quality by Design (AQbD) approach in method development. Optimization step was performed following the Box-Behnken experimental plan and the influence of three critical method parameters (CMPs) towards enantioseparation of the above-mentioned peak pair was examined. CMPs included variation of acetonitrile content in the mobile phase (5-10 %, v/v), pH value of the aqueous phase (6.0-7.0) and ammonium chloride concentration in the aqueous part of the mobile phase (10-30 mmol L-1). The most relevant critical method attributes (CMAs) in this case were the separation criterion between studied critical pair and retention factor of the second eluting peak, (S)-timolol. Qualitative Design Space (DS) was defined by Monte Carlo simulations providing adequate assurance of method's qualitative robustness (π = 95 %). The selected working point situated in the middle of the DS was characterized by following combination of CMPs: acetonitrile content in the mobile phase 7 % (v/v), pH value of the aqueous phase 6.8 and concentration of ammonium chloride in aqueous phase 14 mmol L-1. In the next step, the quantitative robustness was tested by Plackett-Burman experimental design. The validation studies confirmed adequacy of the proposed method for its intended purpose. Finally, Analytical Eco-Scale metric tool was applied to confirm that developed method represents excellent green analytical method compared to the official one.


Assuntos
Ovomucina/química , Timolol/análise , Timolol/isolamento & purificação , Cloreto de Amônio/química , Cromatografia Líquida de Alta Pressão , Limite de Detecção , Modelos Lineares , Modelos Moleculares , Estrutura Molecular , Reprodutibilidade dos Testes , Solventes/química , Estereoisomerismo , Relação Estrutura-Atividade
5.
Chem Commun (Camb) ; 55(100): 15117-15120, 2019 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-31782438

RESUMO

A nitrogen doped bio-carbon catalyst with high specific surface area and a hierarchical interconnected porous structure was fabricated by an in situ gas-foaming strategy from sodium alginate and ammonium chloride. The optimized catalyst displays a fabulous ORR activity, providing a facile approach for the mass production of metal-free bio-carbon catalysts in fuel cells and metal-air batteries.


Assuntos
Carbono/química , Fontes de Energia Elétrica , Gases/química , Nitrogênio/química , Ar , Alginatos/química , Cloreto de Amônio/química , Catálise , Porosidade , Zinco/química
6.
Talanta ; 205: 120082, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31450447

RESUMO

Simultaneous determination of nitrate (NO3‾) and nitrite (NO2‾) in vegetables was performed on a poly(1-vinylimidazole-co-ethylene dimethacrylate) (VIM-EDMA) monolithic column by capillary liquid chromatography (LC) with UV detection. Good linearity (0.5-100 µg mL-1 i.e. 12.5 -2500 µg g-1 in vegetables) was obtained with coefficient of determination > 0.996. Limits of detection (signal-to-noise ratio: S/N= 3) were estimated at 0.06 and 0.05 µg mL-1, which corresponded to 1.50 and 1.25 µg g-1 for NO2‾ and NO3‾, respectively, in vegetables. The limits of quantification (S/N= 10) were estimated at 0.17 and 0.16 µg mL-1 (4.25 and 4.00 µg g-1 in vegetables) for NO2‾ and NO3‾, respectively. Although the detection limits were relatively higher than other LC-UV techniques, this proposed method offered satisfactory sensitivity for complying the Acceptable Daily Intake (ADI) levels set by EU to monitor the occurrence of NO2‾ and NO3‾in vegetables. The intra-day/inter-day precision (0.14-3.35%/0.06-6.93%) and accuracy (90.33-103.32%/96.00-101.26%) were also examined for method validation. No obvious carry-over and decline of separation efficiency were observed for more than 200 analyses of real samples. The occurrence of NO2‾ and NO3‾in various vegetable samples was investigated to demonstrate the potential of utilizing the developed polymeric monolith-based capillary LC-UV method for food safety application.


Assuntos
Cromatografia Líquida/métodos , Análise de Alimentos/métodos , Imidazóis/química , Metacrilatos/química , Nitratos/análise , Nitritos/análise , Polietilenoglicóis/química , Polivinil/química , Verduras/química , Cloreto de Amônio/química , Inocuidade dos Alimentos , Limite de Detecção , Reprodutibilidade dos Testes , Raios Ultravioleta
7.
IET Nanobiotechnol ; 11(8): 909-916, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29155389

RESUMO

Coating gold nanorods (GNRs) with polyelectrolytes is an effective approach to make them biocompatible for potential use in photothermal treatment (PTT) of cancer. The authors report the effect of coating of the GNRs with polystyrene sulphonate (PSS-GNRs) and PSS plus poly di-allyl di-methyl ammonium chloride (PDDAC-GNRs) on its photothermal conversion efficiency (PTE), cellular uptake and subsequently the photothermal induced cytotoxicity in human oral cancer cells (NT8e). Coating of GNRs with PSS led to decrease in PTE by ∼30% and further coating it with PDDAC led to its increase to similar level, with respect to as- prepared GNRs. The cellular uptake of PDDAC-GNRs in cancer cells was double than that for PSS-GNRs. PTT of cancer cells after treatment with 60 pM of either PDDAC-GNRs or PSS-GNRs resulted in cytotoxicty of ∼90%. At higher concentration of 120 pM, while PSS-GNRs showed no further change, for PDDAC-GNR the photothermal induced cytotoxicity decreased to ∼50%. The broadening of longitudinal surface plasmon peak of PDDAC-GNRs and appearance of dark clusters in cells under bright-field microscope suggested intracellular clustering of PDDAC-GNRs. In conclusion, despite high PTE and cellular uptake of PDDAC-GNRs, its intracellular clustering (due to acidic pH ) adversely affect the PTT of cancer cells.


Assuntos
Ouro/química , Neoplasias Bucais/patologia , Nanotubos/química , Polieletrólitos/química , Cloreto de Amônio/química , Linhagem Celular Tumoral , Humanos , Poliestirenos/química , Ressonância de Plasmônio de Superfície , Temperatura
8.
Sci Rep ; 6: 31285, 2016 08 22.
Artigo em Inglês | MEDLINE | ID: mdl-27545761

RESUMO

RNA is a linear polymer of nucleotides linked by a ribose-phosphate backbone. Polymerization of nucleotides occurs in a condensation reaction in which phosphodiester bonds are formed. However, in the absence of enzymes and metabolism there has been no obvious way for RNA-like molecules to be produced and then encapsulated in cellular compartments. We investigated 5'-adenosine monophosphate (AMP) and 5'-uridine monophosphate (UMP) molecules confined in multi-lamellar phospholipid bilayers, nanoscopic films, ammonium chloride salt crystals and Montmorillonite clay, previously proposed to promote polymerization. X-ray diffraction was used to determine whether such conditions imposed a degree of order on the nucleotides. Two nucleotide signals were observed in all matrices, one corresponding to a nearest neighbour distance of 4.6 Å attributed to nucleotides that form a disordered, glassy structure. A second, smaller distance of 3.4 Å agrees well with the distance between stacked base pairs in the RNA backbone, and was assigned to the formation of pre-polymers, i.e., the organization of nucleotides into stacks of about 10 monomers. Such ordering can provide conditions that promote the nonenzymatic polymerization of RNA strands under prebiotic conditions. Experiments were modeled by Monte-Carlo simulations, which provide details of the molecular structure of these pre-polymers.


Assuntos
Nucleotídeos/química , Polímeros/química , Cloreto de Amônio/química , Bentonita/química , Lipídeos/química , Conformação Molecular , Difração de Raios X
9.
J Phycol ; 52(1): 116-24, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26987093

RESUMO

As a promising candidate for biodiesel production, the green alga Chlorella protothecoides can efficiently produce oleaginous biomass and the lipid biosynthesis is greatly influenced by the availability of nitrogen source and corresponding nitrogen assimilation pathways. Based on isotope-assisted kinetic flux profiling (KFP), the fluxes through the nitrogen utilization pathway were quantitatively analyzed. We found that autotrophic C. protothecoides cells absorbed ammonium mainly through glutamate dehydrogenase (GDH), and partially through glutamine synthetase (GS), which was the rate-limiting enzyme of nitrogen assimilation process with rare metabolic activity of glutamine oxoglutarate aminotransferase (GOGAT, also known as glutamate synthase); whereas under heterotrophic conditions, the cells adapted to GS-GOGAT cycle for nitrogen assimilation in which GS reaction rate was associated with GOGAT activity. The fact that C. protothecoides chooses the adenosine triphosphate-free and less ammonium-affinity GDH pathway, or alternatively the energy-consuming GS-GOGAT cycle with high ammonium affinity for nitrogen assimilation, highlights the metabolic adaptability of C. protothecoides exposed to altered nitrogen conditions.


Assuntos
Chlorella/crescimento & desenvolvimento , Chlorella/metabolismo , Modelos Biológicos , Nitrogênio/metabolismo , Cloreto de Amônio/química , Cloreto de Amônio/metabolismo , Processos Autotróficos , Glutamato Desidrogenase/metabolismo , Glutamato-Amônia Ligase/metabolismo , Processos Heterotróficos , Cinética , Redes e Vias Metabólicas , Isótopos de Nitrogênio/análise , Isótopos de Nitrogênio/metabolismo
10.
Waste Manag ; 48: 638-643, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26445362

RESUMO

The aim of the study was focused on extraction of fleshing oil from limed fleshings with different neutralization process by ammonium chloride (NH4Cl) and hydrochloric acid (HCl) followed by solvent extraction. The production of fatty acid methyl esters (FAMEs) from limed fleshing oil by two stage process has also been investigated. The central composite design (CCD) was used to study the effect of process variables viz., amount of flesh, particle size and time of fleshing oil extraction. The maximum yield of fleshing oil from limed fleshings post neutralization by ammonium chloride (NH4Cl) and hydrochloric acid (HCl) was 26.32g and 12.43g obtained at 200g of flesh, with a particle size of 3.90mm in the time period of 2h. Gas chromatography analysis reveals that the biodiesel (FAME) obtained from limed fleshings is rich in oleic and palmitic acids with weight percentages 46.6 and 32.2 respectively. The resulting biodiesel was characterized for its physio-chemical properties of diesel as per international standards (EN14214).


Assuntos
Biocombustíveis , Compostos de Cálcio/química , Fontes Geradoras de Energia , Óxidos/química , Cloreto de Amônio/química , Sulfato de Amônio/química , Cromatografia Gasosa , Ésteres/química , Ácido Clorídrico/química , Concentração de Íons de Hidrogênio , Ácido Oleico/química , Ácido Palmítico/química , Tamanho da Partícula , Análise de Regressão , Ácidos Sulfúricos/química , Temperatura
11.
J Hazard Mater ; 298: 162-9, 2015 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-26051992

RESUMO

Pyrolyzed biochars from an industrial medicinal residue waste were modified by microwave activation and NH4Cl impregnation. Mercury adsorption of different modified biochars was investigated in a quartz fixed-bed reactor. The results indicated that both physisorption and chemisorption of Hg(0) occurred on the surface of M6WN5 which was modified both microwave and 5wt.% NH4Cl loading, and exothermic chemisorption process was a dominant route for Hg(0) removal. Microwave activation improved pore properties and NH4Cl impregnation introduced good active sites for biochars. The presence of NO and O2 increased Hg(0) adsorption whereas H2O inhibited Hg(0) adsorption greatly. A converse effect of SO2 was observed on Hg(0) removal, namely, low concentration of SO2 promoted Hg(0) removal obviously whereas high concentration of SO2 suppressed Hg(0) removal. The Hg(0) removal by M6WN5 was mainly due to the reaction of the C−Cl with Hg(0) to form HgCl2, and the active state of C−Cl(*) groups might be an intermediate group in this process. Thermodynamic analysis showed that mercury adsorption by the biochars was exothermic process and apparent adsorption energy was 43.3 kJ/mol in the range of chemisorption. In spite of low specific surface area, M6WN5 proved to be a promising Hg(0) sorbent in flue gas when compared with other sorbents.


Assuntos
Carvão Vegetal , Resíduos de Drogas/química , Resíduos de Serviços de Saúde/análise , Mercúrio/isolamento & purificação , Adsorção , Cloreto de Amônio/química , Gases , Micro-Ondas , Óxido Nítrico/química , Oxigênio/química , Dióxido de Enxofre/química , Temperatura , Termodinâmica
12.
J Leukoc Biol ; 97(6): 1121-31, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25877929

RESUMO

Neutrophils kill microorganisms by inducing exocytosis of granules with antibacterial properties. Four isoforms of the "a" subunit of V-ATPase-a1V, a2V, a3V, and a4V-have been identified. a2V is expressed in white blood cells, that is, on the surface of monocytes or activated lymphocytes. Neutrophil associated-a2V was found on membranes of primary (azurophilic) granules and less often on secondary (specific) granules, tertiary (gelatinase granules), and secretory vesicles. However, it was not found on the surface of resting neutrophils. Following stimulation of neutrophils, primary granules containing a2V as well as CD63 translocated to the surface of the cell because of exocytosis. a2V was also found on the cell surface when the neutrophils were incubated in ammonium chloride buffer (pH 7.4) a weak base. The intracellular pH (cytosol) became alkaline within 5 min after stimulation, and the pH increased from 7.2 to 7.8; this pH change correlated with intragranular acidification of the neutrophil granules. Upon translocation and exocytosis, a2V on the membrane of primary granules remained on the cell surface, but myeloperoxidase was secreted. V-ATPase may have a role in the fusion of the granule membrane with the cell surface membrane before exocytosis. These findings suggest that the granule-associated a2V isoform has a role in maintaining a pH gradient within the cell between the cytosol and granules in neutrophils and also in fusion between the surface and the granules before exocytosis. Because a2V is not found on the surface of resting neutrophils, surface a2V may be useful as a biomarker for activated neutrophils.


Assuntos
Grânulos Citoplasmáticos/imunologia , Neutrófilos/imunologia , ATPases Vacuolares Próton-Translocadoras/imunologia , Cloreto de Amônio/química , Movimento Celular/efeitos dos fármacos , Citocalasina D/farmacologia , Grânulos Citoplasmáticos/efeitos dos fármacos , Grânulos Citoplasmáticos/metabolismo , Exocitose/efeitos dos fármacos , Regulação da Expressão Gênica , Humanos , Concentração de Íons de Hidrogênio , Membranas Intracelulares/efeitos dos fármacos , Membranas Intracelulares/imunologia , Membranas Intracelulares/metabolismo , Isoenzimas/genética , Isoenzimas/imunologia , Fusão de Membrana/efeitos dos fármacos , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Ativação de Neutrófilo/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Peroxidase/genética , Peroxidase/imunologia , Peroxidase/metabolismo , Cultura Primária de Células , Transdução de Sinais , Tetraspanina 30/genética , Tetraspanina 30/imunologia , ATPases Vacuolares Próton-Translocadoras/genética
13.
J Phys Chem B ; 118(25): 7012-25, 2014 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-24877990

RESUMO

The basic requirement for understanding the nonviral gene delivery pathway is a thorough biophysical characterization of DNA polyplexes. In this work, we have studied the interactions between calf-thymus DNA (ctDNA)and a new series of linear cationic block copolymers (BCPs). The BCPs were synthesized via controlled radical polymerization using [3-(methacryloylamino)propyl] -trimethylammonium chloride (MAPTAC) and poly(ethyleneglycol) methyl ether (PEGMe) as comonomers. UV−visible spectroscopy, ethidium bromide dye exclusion, and gel electrophoresis study revealed that these cationic BCPs were capable of efficiently binding with DNA. Steady-state fluorescence, UV melting, gel electrophoresis, and circular dichroism results suggested increased binding for BCPs containing higher PEG. Hydrophobic interactions between the PEG and the DNA base pairs became significant at close proximity of the two macromolecules, thereby influencing the binding trend. DLS studies showed a decrease in the size of DNA molecules at lower charge ratio (the ratio of "+" charge of the polymer to "−" charge of DNA) due to compaction, whereas the size increased at higher charge ratio due to aggregation among the polyplexes. Additionally, we have conducted kinetic studies of the binding process using the stop-flow fluorescence method. All the results of BCP−DNA binding studies suggested a two-step reaction mechanism--a rapid electrostatic binding between the cationic blocks and DNA, followed by a conformational change of the polyplexes in the subsequent step that led to DNA condensation. The relative rate constant(k'(1)) of the first step was much higher compared to that of the second step (k'(2)), and both were found to increase with an increase in BCP concentration. The charge ratios as well as the PEG content in the BCPs had a marked effect on the kinetics of the DNA−BCP polyplex formation. Introduction of a desired PEG chain length in the synthesized cationic blocks renders them potentially useful as nonviral gene delivery agents.


Assuntos
DNA/química , Polietilenoglicóis/química , Polímeros/química , Cloreto de Amônio/química , Animais , Cátions/química , Bovinos , Etídio/química , Cinética , Metacrilatos/química , Transição de Fase , Polimerização , Polímeros/síntese química
14.
Chemistry ; 20(12): 3273-6, 2014 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-24677187

RESUMO

In nature, diatoms and sponges are exquisite examples of well-defined structures produced by silica biomineralisation, in which proteins play an important role. However, the artificial peptide templating route for the silica mesostructure remains a formidable and unsolved challenge. Herein, we report our effort on the design of amphiphilic peptides for synthesising a highly ordered two-dimensional (2D)-hexagonal and lamellar chiral silica mesostructure using trimethoxysilylpropyl-N,N,N-trimethylammonium chloride as the co-structure directing agent (CSDA). The geometry of the peptide was designed by adding proline residues into the hydrophobic chain of the peptide to break the b-sheet conformation by weakening the intermolecular hydrogen bonds; this led to the mesophase transformation from the most general lamellar structure to the 2D hexagonal P6mm mesostructure by increasing the amphiphilic molecules packing parameter g. Enantiomerically pure chiral mesostructures were formed thanks to the intrinsic chirality and relatively strong intermolecular hydrogen bonds of peptides.


Assuntos
Cloreto de Amônio/química , Peptídeos/química , Dióxido de Silício/química , Compostos de Trimetil Amônio/química , Compostos de Trimetilsilil/química , Biomimética , Ligação de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Porosidade , Estereoisomerismo
15.
Waste Manag ; 33(12): 2641-52, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24035244

RESUMO

Recycling biowaste digestates on agricultural land diverts biodegradable waste from landfill disposal and represents a sustainable source of nutrients and organic matter (OM) to improve soil for crop production. However, the dynamics of nitrogen (N) release from these organic N sources must be determined to optimise their fertiliser value and management. This laboratory incubation experiment examined the effects of digestate type (aerobic and anaerobic), waste type (industrial, agricultural and municipal solid waste or sewage sludge) and soil type (sandy loam, sandy silt loam and silty clay) on N availability in digestate-amended soils and also quantified the extent and significance of the immobilisation of N within the soil microbial biomass, as a possible regulatory mechanism of N release. The digestate types examined included: dewatered, anaerobically digested biosolids (DMAD); dewatered, anaerobic mesophilic digestate from the organic fraction of municipal solid waste (DMADMSW); liquid, anaerobic co-digestate of food and animal slurry (LcoMAD) and liquid, thermophilic aerobic digestate of food waste (LTAD). Ammonium chloride (NH4Cl) was included as a reference treatment for mineral N. After 48 days, the final, maximum net recoveries of mineral N relative to the total N (TN) addition in the different digestates and unamended control treatments were in the decreasing order: LcoMAD, 68%; LTAD, 37%, DMAD, 20%; and DMADMSW, 11%. A transient increase in microbial biomass N (MBN) was observed with LTAD application, indicating greater microbial activity in amended soil and reflecting the lower stability of this OM source, compared to the other, anaerobic digestate types, which showed no consistent effects on MBN compared to the control. Thus, the overall net release of digestate N in different soil types was not regulated by N transfer into the soil microbial biomass, but was determined primarily by digestate properties and the capacity of the soil type to process and turnover digestate N. In contrast to the sandy soil types, where nitrate (NO3-) concentrations increased during incubation, there was an absence of NO3- accumulation in the silty clay soil amended with LTAD and DMADMSW. This provided indirect evidence for denitrification activity and the gaseous loss of N, and the associated increased risk of greenhouse gas emissions under certain conditions of labile C supply and/or digestate physical structure in fine-textured soil types. The significance and influence of the interaction between soil type and digestate stability and physical properties on denitrification processes in digestate-amended soils require urgent investigation to ensure management practices are appropriate to minimise greenhouse gas emissions from land applied biowastes.


Assuntos
Gases/análise , Ciclo do Nitrogênio , Nitrogênio/química , Solo/química , Resíduos/análise , Aerobiose , Agricultura , Cloreto de Amônio/química , Anaerobiose , Biomassa , Efeito Estufa
16.
Phys Chem Chem Phys ; 15(27): 11221-5, 2013 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-23749028

RESUMO

Mesoporous SnO2 with a high surface area of 292.7 m(2) g(-1) has been successfully synthesized via a low-cost NH4Cl-based ionothermal route. When evaluated as a gas sensor, impressive performances towards N2H4 and HCHO are achieved owing to its excellent chemical reactivity towards oxygen.


Assuntos
Cloreto de Amônio/química , Nanoestruturas/química , Oxigênio/química , Temperatura , Compostos de Estanho/síntese química , Amônia/química , Formaldeído/química , Hidrazinas/química , Metanol/química , Oxirredução , Porosidade , Propriedades de Superfície , Compostos de Estanho/química
17.
Methods Mol Biol ; 946: 267-83, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23179838

RESUMO

Hematopoietic stem cells (HSCs) present in small numbers in adult bone marrow (BM), peripheral blood (PB) and umbilical cord blood (CB) produce a heterogeneous pool of progenitors that can be detected in vitro using colony forming cell (CFC) assays. Hematopoietic progenitor cells proliferate and differentiate to produce colonies of maturing cells when cultured in a semisolid methylcellulose-based medium that is supplemented with suitable growth factors and other supplements. The colonies are then classified and enumerated in situ by light microscopy or an automated imaging instrument. CFC assays are important tools in basic hematology research but are also used by clinical cell processing laboratories to measure the progenitor cell content of BM, CB and mobilized PB (MPB) preparations used for cell transplantation. Standard CFC assays for human progenitor cells require a culture period of at least 14 days to enable optimal outgrowth and differentiation of the maximum number of CFCs in a cell preparation. In this chapter protocols are described for the detection and enumeration of myeloid multipotential progenitors and committed progenitors of the erythroid, monocyte, and granulocyte lineages in samples from human PB, MPB, BM, and CB. In addition protocols are described for a modified version of the CFC-assay that allows accurate enumeration of total CFC numbers in CB or MPB after a culture period of only 7 days, but without distinction of colony types.


Assuntos
Ensaio de Unidades Formadoras de Colônias/métodos , Células-Tronco Hematopoéticas/citologia , Cloreto de Amônio/química , Contagem de Células , Separação Celular , Meios de Cultura/química , Células Eritroides/citologia , Humanos , Megacariócitos/citologia , Metilcelulose/química , Suspensões , Fatores de Tempo
18.
PLoS Pathog ; 8(5): e1002694, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22589725

RESUMO

Disparate enveloped viruses initiate infection by fusing with endosomes. However, the highly diverse and dynamic nature of endosomes impairs mechanistic studies of fusion and identification of sub-cellular sites supporting the nucleocapsid release. We took advantage of the extreme stability of avian retrovirus-receptor complexes at neutral pH and of acid-dependence of virus-endosome fusion to isolate the latter step from preceding asynchronous internalization/trafficking steps. Viruses were trapped within endosomes in the presence of NH4Cl. Removal of NH4Cl resulted in a quick and uniform acidification of all subcellular compartments, thereby initiating synchronous viral fusion. Single virus imaging demonstrated that fusion was initiated within seconds after acidification and often culminated in the release of the viral core from an endosome. Comparative studies of cells expressing either the transmembrane or GPI-anchored receptor isoform revealed that the transmembrane receptor delivered the virus to more fusion-permissive compartments. Thus the identity of endosomal compartments, in addition to their acidity, appears to modulate viral fusion. A more striking manifestation of the virus delivery to distinct compartments in the presence of NH4Cl was the viral core release into the cytosol of cells expressing the transmembrane receptor and into endosomes of cells expressing the GPI-anchored isoform. In the latter cells, the newly released cores exhibited restricted mobility and were exposed to a more acidic environment than the cytoplasm. These cores appear to enter into the cytosol after an additional slow temperature-dependent step. We conclude that the NH4Cl block traps the virus within intralumenal vesicles of late endosomes in cells expressing the GPI-anchored receptor. Viruses surrounded by more than one endosomal membrane release their core into the cytoplasm in two steps--fusion with an intralumenal vesicle followed by a yet unknown temperature-dependent step that liberates the core from late endosomes.


Assuntos
Vírus do Sarcoma Aviário/genética , Vírus do Sarcoma Aviário/metabolismo , Endossomos/virologia , Proteínas do Core Viral/metabolismo , Proteínas Virais de Fusão/metabolismo , Cloreto de Amônio/química , Animais , Compartimento Celular , Linhagem Celular , Chlorocebus aethiops , Endossomos/metabolismo , Células HEK293 , Humanos , Isoformas de Proteínas/biossíntese , Transporte Proteico , Proteínas dos Retroviridae/genética , Proteínas dos Retroviridae/metabolismo , Proteínas do Core Viral/genética , Proteínas Virais de Fusão/genética , Internalização do Vírus
19.
Artigo em Inglês | MEDLINE | ID: mdl-22591796

RESUMO

The absorption properties of zinc-nickel alloy electrolyte were studied by visible spectrophotometer. The results show that the relationship between the absorbance of the zinc-nickel alloy electrolyte and Ni(2+) concentration in the electrolyte obeys Beer's law at 660 nm. In addition, other components except Ni(2+) in the zinc-nickel alloy electrolyte such as zinc chloride, ammonium chloride, potassium chloride and boric acid have no obvious effect on the absorbance of zinc-nickel alloy electrolyte. Based on these properties, a new method is developed to determine Ni(2+) concentration in zinc-nickel alloy electrolyte. Comparing with other methods, this method is simple, direct and accurate. Moreover, the whole testing process does not consume any reagent and dilution, and after testing, the electrolyte samples can be reused without any pollution to the environment.


Assuntos
Ligas/química , Eletrólitos/química , Níquel/análise , Espectrofotometria/métodos , Absorção , Cloreto de Amônio/química , Ácidos Bóricos/química , Cloretos/química , Concentração de Íons de Hidrogênio , Modelos Lineares , Cloreto de Potássio/química , Temperatura , Compostos de Zinco/química
20.
Biomaterials ; 33(5): 1682-7, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22123601

RESUMO

Magnetic carriers with efficient loading, delivery, and release of drugs are required for magnetically guided drug delivery system (DDS) as the potential cancer therapy. The present article describes the fabrication of porous FePt capsules approximately 340 nm in diameter with large pores of 20 nm in an ultrathin shell of 10 nm and demonstrates their application to a magnetically guided DDS in vitro. An aqueous anti-cancer drug is easily introduced in the hollow space of the capsules without external stimuli and released to cancer cells on cue through the magnetic shell composed of an ordered-alloy FePt network structure, which exhibits superparamagnetic features at approximately body temperature. The drug-loaded magnetic capsules coated with a lipid membrane are efficiently guided to the cancer cells within 15 min using a NdFeB magnet (0.2 T), and more than 70% of the cancer cells are destroyed.


Assuntos
Antineoplásicos/farmacologia , Sistemas de Liberação de Medicamentos/métodos , Ferro/química , Magnetismo/métodos , Platina/química , Cloreto de Amônio/química , Cápsulas , Linhagem Celular Tumoral , Doxorrubicina/farmacologia , Humanos , Lipídeos/química , Microscopia de Fluorescência , Nanopartículas/química , Nanopartículas/ultraestrutura , Porosidade/efeitos dos fármacos , Dióxido de Silício/química , Difração de Raios X
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