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1.
Ecotoxicol Environ Saf ; 266: 115603, 2023 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-37856986

RESUMEN

Hydroxylamine is a highly reactive inorganic nitrogen compound that not only has a toxic effect on microorganisms, but also makes wastewater treatment more difficult, which in turn damages the environment and even endangers human health. This study reported a new method for converting of hydroxylamine by adding sodium carbonate or calcium bicarbonate to the hydroxylamine-polluted wastewater. The conversion efficiency of hydroxylamine was more than 99% in the presence of sodium carbonate or calcium bicarbonate under the reaction conditions of 25 °C, C/N ratio 15, and dissolved oxygen 7.4 mg/L. And its maximal conversion rate can reach 3.49 mg/L/h. This method overcomes various shortcomings of the reported hydroxylamine removal technologies that require a large material dosage and high cost. The technology in this report has many advantages: low cost, 'green' environmental protection, easy market promotion, and high economic benefits.


Asunto(s)
Hidroxilaminas , Aguas Residuales , Humanos , Hidroxilamina , Suplementos Dietéticos , Nitrógeno , Carbonato de Calcio
2.
Environ Sci Pollut Res Int ; 30(32): 79282-79296, 2023 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-37284959

RESUMEN

Rational treatment of drinking water treatment residues (WTR) has become an environmental and social issue due to the risk of secondary contamination. WTR has been commonly used to prepare adsorbents because of its clay-like pore structure, but then requires further treatment. In this study, a Fenton-like system of H-WTR/HA/H2O2 was constructed to degrade organic pollutants in water. Specifically, WTR was modified by heat treatment to increase its adsorption active site, and to accelerate Fe(III)/Fe(II) cycling on the catalyst surface by the addition of hydroxylamine (HA). Moreover, the effects of pH, HA and H2O2 dosage on the degradation were discussed with methylene blue (MB) as the target pollutant. The mechanism of the action of HA was analyzed and the reactive oxygen species in the reaction system were determined. Combined with the reusability and stability experiments, the removal efficiency of MB remained 65.36% after 5 cycles. Consequently, this study may provide new insights into the resource utilization of WTR.


Asunto(s)
Agua Potable , Contaminantes Químicos del Agua , Hidroxilamina/química , Peróxido de Hidrógeno/química , Hierro/química , Azul de Metileno , Calor , Contaminantes Químicos del Agua/análisis , Hidroxilaminas , Oxidación-Reducción
3.
Appl Environ Microbiol ; 88(7): e0241621, 2022 04 12.
Artículo en Inglés | MEDLINE | ID: mdl-35285242

RESUMEN

In the nitrogen (N) cycle, nitrogenous compounds are chemically and biologically converted to various aqueous and gaseous N species. The 15N-labeling approach is a powerful culture-dependent technique to obtain insights into the complex nitrogen transformation reactions that occur in cultures. In the 15N-labeling approach, the fates of supplemented 15N- and/or unlabeled gaseous and aqueous compounds are tracked by mass spectrometry (MS) analysis, whereas MS analysis of aqueous N species requires laborious sample preparation steps and is performed using isotope-ratio mass spectrometry, which requires an expensive mass spectrometer. We developed a simple and high-throughput MS method for determining the 15N atoms percent of NH4+, NO2-, NO3-, NH2OH, and N2H4, where liquid samples (<0.5 mL) were mixed with colorimetric reagents (naphthylethylenediamine for NO2-, indophenol for NH4+, and p-aminobenzaldehyde for N2H4), and the mass spectra of the formed N complex dyes were obtained by matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) MS. NH2OH and NO3- were chemically converted to NO2- by iodine oxidation and copper/hydrazine reduction reaction, respectively, prior to the above colorimetric reaction. The intensity of the isotope peak (M + 1 or M + 2) increased when the N complex dye was formed by coupling with a 15N-labeled compound, and a linear relationship was found between the determined 15N/14N peak ratio and 15N atom% for the tested N species. The developed method was applied to bacterial cultures to examine their N-transformation reactions, enabling us to observe the occurrence of NO2- oxidation and NO3- reduction in a hypoxic Nitrobacter winogradskyi culture. IMPORTANCE15N/14N analysis for aqueous N species is a powerful tool for obtaining insights into the global N cycle, but the procedure is cumbersome and laborious. The combined use of colorimetric reagents and MALDI-TOF MS, designated color MALDI-TOF MS, enabled us to determine the 15N atom% of common aqueous N species without laborious sample preparation and chromatographic separation steps; for instance, the 15N atom% of NO2- can be determined from >1,000 liquid samples daily at <$1 (U.S.) per 384 samples for routine analysis. This convenient MS method is a powerful tool that will advance our ability to explore the N-transformation reactions that occur in various environments and biological samples.


Asunto(s)
Nitratos , Nitritos , Colorimetría , Hidrazinas , Hidroxilamina , Isótopos , Rayos Láser , Nitrógeno , Dióxido de Nitrógeno , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos
4.
Sci Total Environ ; 794: 148478, 2021 Nov 10.
Artículo en Inglés | MEDLINE | ID: mdl-34217093

RESUMEN

Hydroxylamine (NH2OH) as the putative intermediate for anammox ensures the robustness of partial nitritation/anammox (PN/A) process; however, the feasible for NH2OH addition to improve the stability of PN/A process under low-strength ammonia (NH4+-N) condition need to be further investigated. In this study, the restoration and steady operation of mainstream PN/A process were investigated to treat real sewage with in situ NH2OH added in a continuous alternating anoxic/aerobic with integrated fixed-film activated sludge (A3-IFAS) reactor. Results showed that the deteriorated PN/A process caused by nitrate (NO3--N) built-up was rapidly restored with a distinct decrease of the NO3--Nproduced/NH4+-Nconsumed ratio from 28.7% to <10.0% within 20 days, after 5 mg N/L of NH2OH was added daily into the aerobic zone of A3-IFAS reactor. After 230 days of operation, the average total nitrogen (TN) and phosphate (PO43--P) removal efficiencies of 80.8% and 91.5%, respectively were stably achieved, with average effluent sCOD, NH4+-N, TN and PO43--P concentrations reaching 23.1, 2.3, 7.7 and 0.4 mg/L, respectively. Microbial community characterization revealed Candidatus Brocadia (3.60% and 2.92%) and Ignavibacteriae (1.56% and 2.66%) as the dominant anammox bacteria and denitrifying bacteria, respectively, jointly attached in the biofilm_1 and biofilm_2, while Candidatus Microthrix (5.17%) dominant in floc sludge was main responsible for phosphorus removal. This study confirmed that NH2OH addition is an effective strategy for nitrite-oxidizing bacteria suppression, contributing to the in situ restoration of PN/A process and high stable mainstream nitrogen and phosphorus removal in a continuous PN/A process from real sewage.


Asunto(s)
Compuestos de Amonio , Aguas del Alcantarillado , Reactores Biológicos , Desnitrificación , Hidroxilamina , Hidroxilaminas , Nitrógeno , Oxidación-Reducción , Fósforo , Aguas Residuales
5.
Int J Nanomedicine ; 15: 2935-2945, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32425525

RESUMEN

BACKGROUND: New anticancer agents that rely on natural/healthy, not synthetic/toxic, components are very much needed. METHODS: Ricinoleyl hydroxamic acid (RHA) was synthesized from castor oil and hydroxylamine using Lipozyme TL IM as a catalyst. To optimize the conversion, the effects of the following parameters were investigated: type of organic solvent, period of reaction, amount of enzyme, the molar ratio of reactants and temperature. The highest conversion was obtained when the reaction was carried out under the following conditions: hexane as a solvent; reaction period of 48 hours; 120 mg of Lipozyme TL IM/3 mmol oil; HA-oil ratio of 19 mmol HA/3 mmol oil; and temperature of 40°C. The cytotoxicity of the synthesized RHA was assessed using human dermal fibroblasts (HDF), and its application towards fighting cancer was assessed using melanoma and glioblastoma cancer cells over a duration of 24 and 48 hours. RESULTS: RHA was successfully synthesized  and it demonstrated strong anticancer activity against glioblastoma and melanoma cells at as low as a 1 µg/mL concentration while it did not demonstrate any toxicity against HDF cells. CONCLUSION: This is the first report on the synthesis of RHA with great potential to be used as a new anticancer agent.


Asunto(s)
Antineoplásicos/síntesis química , Antineoplásicos/farmacología , Aceite de Ricino/química , Neoplasias Encefálicas/tratamiento farmacológico , Neoplasias Encefálicas/patología , Catálisis , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Glioblastoma/tratamiento farmacológico , Glioblastoma/patología , Hexanos/química , Humanos , Hidroxilamina/química , Lipasa/química , Lipasa/metabolismo , Melanoma/tratamiento farmacológico , Melanoma/patología , Solventes/química
6.
Water Sci Technol ; 73(3): 493-500, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26877030

RESUMEN

Cuprous copper [Cu(I)] reacts with sodium persulfate (PDS) to generate sulfate radical SO4(-)•, but it has been seldom investigated owing to its instability and difficulty in dissolving it. This study proposes a new method to regenerate Cu(I) from cupric copper [Cu(II)] by addition of hydroxylamine (HA) to induce the continuous production of radicals through active PDS, and investigates the resulting enhanced methyl orange (MO) degradation efficiency and mechanism in the new system. HA accelerated the degradation of MO markedly in the pH range from 6.0 to 8.0 in the HA/Cu(II)/PDS process. Both SO4(-)• and hydroxyl radicals (•OH) were considered as the primary reactive radicals in the process. The MO degradation in the HA/Cu(II)/PDS process can be divided into three stages: the fast stage, the transitory stage, and the low stage. MO degradation was enhanced with increased dosage of PDS. Although high dosage of HA could accelerate the transformation of the Cu(II)/Cu(I) cycle to produce more reactive radicals, excess HA can quench the reactive radicals. This study indicates that through a copper-redox cycling mechanism by HA, the production of SO4(-)• and •OH can be strongly enhanced, and the effective pH range can be expanded to neutral conditions.


Asunto(s)
Cobre/química , Hidroxilamina/química , Compuestos de Sodio/química , Sulfatos/química , Compuestos Azo/química , Concentración de Iones de Hidrógeno , Oxidación-Reducción
7.
New Phytol ; 206(1): 118-126, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25615266

RESUMEN

Hydrogen peroxide (H2O2) has been reported to increase lignin formation, enhance cell wall rigidification, restrict cell expansion and inhibit root elongation. However, our results showed that it not only inhibited rice (Oryza sativa) root elongation, but also increased root diameter. No study has reported how and why H2O2 increases cell expansion and root diameter. Exogenous H2O2 and its scavenger 4-hydroxy-Tempo were applied to confirm the roles of H2O2. Immunofluorescence, fluorescence probe, ruthenium red staining, histological section and spectrophotometry were used to monitor changes in the degree of pectin methylesterification, pectin content, pectin methylesterase (PME) activity and H2O2 content. Exogenous H2O2 inhibited root elongation, but increased cell expansion and root diameter significantly. H2O2 not only increased the region of pectin synthesis and pectin content in root tips, but also increased PME activity and pectin demethylesterification. The scavenger 4-hydroxy-Tempo reduced root H2O2 content and recovered H2O2-induced increases in cell expansion and root diameter by inhibiting pectin synthesis, PME activity and pectin demethylesterification. H2O2 plays a novel role in the regulation of pectin synthesis, PME activity and pectin demethylesterification. H2O2 increases cell expansion and root diameter by increasing pectin content and demethylesterification.


Asunto(s)
Hidrolasas de Éster Carboxílico/metabolismo , Regulación de la Expresión Génica de las Plantas , Peróxido de Hidrógeno/farmacología , Oryza/fisiología , Pectinas/metabolismo , Metabolismo de los Hidratos de Carbono , Hidrolasas de Éster Carboxílico/genética , Pared Celular/metabolismo , Óxidos N-Cíclicos/farmacología , Esterificación , Peróxido de Hidrógeno/metabolismo , Hidroxilamina/farmacología , Meristema/efectos de los fármacos , Meristema/enzimología , Meristema/fisiología , Oryza/efectos de los fármacos , Oryza/enzimología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Cápsula de Raíz de Planta/efectos de los fármacos , Cápsula de Raíz de Planta/enzimología , Cápsula de Raíz de Planta/fisiología , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/enzimología , Raíces de Plantas/fisiología , Especies Reactivas de Oxígeno/metabolismo
8.
Mol Med Rep ; 11(2): 837-42, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25374015

RESUMEN

The present study aimed to investigate the role of nitric oxide (NO) against perinatal hypoxic­ischemic brain damage (HIBD) in rats by electroacupuncture (EA) and to examine its potential neuroprotective mechanism. NO content, the number of positive cells, neuronal nitric oxide synthase (nNOS) and nuclear factor­κB (NF­κB) in rat cortex cells were determined. The results demonstrated that treatment with EA significantly downregulated the NO content in the cortex cells (*P<0.05, **P<0.01, compared with the control groups) and alleviated cell damage in the cortex of rats with HIBD. The activator, S­adenosyl­L­methionine and the inhibitor, hydroxylamine of cystathionine­ß­synthase (CBS), aggravated and remitted the hypoxic damage in the cortex cells, respectively. In addition, treatment with EA significantly downregulated the expression of nNOS and NF­κB in the rat cortex cells (*P<0.05, **P<0.01, compared with the control groups). The results also indicated that treatment with EA downregulated the NO content of cortical cells against HIBD via the NF­κB/nNOS pathway and further implied that the hydrogen sulfide/CBS system may be involved in the process. The present study provided a significant reference for the prevention and treatment of HIBD using the EA technique and also described a novel protective mechanism.


Asunto(s)
Hipoxia-Isquemia Encefálica/patología , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo I/metabolismo , Óxido Nítrico/metabolismo , Animales , Corteza Cerebral/enzimología , Corteza Cerebral/metabolismo , Cistationina betasintasa/antagonistas & inhibidores , Cistationina betasintasa/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Electroacupuntura , Sulfuro de Hidrógeno/metabolismo , Hidroxilamina/farmacología , Hipoxia-Isquemia Encefálica/metabolismo , Ratas , Ratas Sprague-Dawley , S-Adenosilmetionina/farmacología , Regulación hacia Arriba/efectos de los fármacos
9.
Analyst ; 139(15): 3796-803, 2014 Aug 07.
Artículo en Inglés | MEDLINE | ID: mdl-24899364

RESUMEN

A highly sensitive and selective chemiluminescent (CL) biosensor for adenosine triphosphate (ATP) was developed by taking advantage of the ATP-dependent enzymatic reaction (ATP-DER), the powerful signal amplification capability of rolling circle amplification (RCA), and hydroxylamine-amplified gold nanoparticles (Au NPs). The strategy relies on the ability of ATP, a cofactor of T4 DNA ligase, to trigger the ligation-RCA reaction. In the presence of ATP, the T4 DNA ligase catalyzes the ligation reaction between the two ends of the padlock probe, producing a closed circular DNA template that initiates the RCA reaction with phi29 DNA polymerase and dNTP. Therein, many complementary copies of the circular template can be generated. The ATP-DER is eventually converted into a detectable CL signal after a series of processes, including gold probe hybridization, hydroxylamine amplification, and oxidative gold metal dissolution coupled with a simple and sensitive luminol CL reaction. The CL signal is directly proportional to the ATP level. The results showed that the detection limit of the assay is 100 pM of ATP, which compares favorably with those of other ATP detection techniques. In addition, by taking advantage of ATP-DER, the proposed CL sensing system exhibits extraordinary specificity towards ATP and could distinguish the target molecule ATP from its analogues. The proposed method provides a new and versatile platform for the design of novel DNA ligation reaction-based CL sensing systems for other cofactors. This novel ATP-DER based CL sensing system may find wide applications in clinical diagnosis as well as in environmental and biomedical fields.


Asunto(s)
Adenosina Trifosfato/sangre , Oro/química , Hidroxilamina/química , Mediciones Luminiscentes/métodos , Nanopartículas del Metal/química , Adenosina Trifosfato/análisis , Técnicas Biosensibles/métodos , Humanos , Límite de Detección , Luminol/análisis , Técnicas de Amplificación de Ácido Nucleico/métodos
10.
Explore (NY) ; 10(2): 99-108, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24607076

RESUMEN

PREMISE: Applied Kinesiology (AK) is a diagnostic technique widely used within the Integrative Medical community. In essence, it posits that a question can be mentally held in a person's mind, sometimes while they are holding a substance like a vitamin, or a food sample, and by measuring relative muscular weakness an answer as to whether the substance or the condition represented by the question is good for that person can be obtained. This AK is presumed to have a diagnostic capability. That being presumed, this study asks the following questions: (1) Is there a difference in muscular strength when an individual holds a substance that is inimical to life processes (a poison solution), as compared to a substance that is essential for life (normal saline)? (2) Is this effect a transaction involving input from both the person being measured and the kinesiologist doing the measurement or is it only the person being measured? (3) As an extension of question 2, is the result the same when different kinesiologists take the measurement or when no kinesiologist is involved? (4) Does belief, expectation, gender, or time cognition play a role in determining response? METHODOLOGY: To answer these questions, which would help to define the parameters of the AK process, 51 participants were tested during three trials each, first by one kinesiologist, then by another, and finally, with no kinesiologist present by grip strength indicated using a hand dynamometer. Grip strength being a self-administered AK test of relative muscular strength. For each trial, a pair of randomly numbered sealed vials, each pair in a randomly numbered plastic bag, were used as the objects of the trial. In each bag, one vial contained saline solution while the other was filled with a slightly smaller amount of saline solution to which had been added ionic hydroxylamine hydrochloride (NH3OH)(+), producing a toxic solution of 9mg/ml. Each trial consisted of a separate muscle test for each vial. All present at the trials were blind as to which vial contained the toxin. And all who prepared the vials were blind to the trials. The force used by the kinesiologists in each of their trials was measured via a pressure pad system. The hand dynamometer trials were conducted with no kinesiologist present. RESULTS: Of the 151 sets of trials, the toxic vial was identified correctly in 80 of them (53%), resulting in a one-tailed exact binomial P-value of .258. Results for two of the kinesiologists were almost exactly at chance. For the third kinesiologist, there was a one-tailed exact binomial P-value of .18 (unadjusted for multiple testing). Results for the dynamometer were also almost exactly at chance. Testing whether there was a significant difference in proportions for whom the AK test worked based on belief about whether it would work resulted in non-significant χ(2) values of 0.6 (P = .439) for the trials with one kinesiologist and 2.222 (P = .136) for the hand dynamometer trials. The final variable examined was gender. While there was no significant difference in performance for males and females for the trials of the male kinesiologist or the hand dynamometer, the combined data for the two female kinesiologists did reveal a difference. Of the 33 sessions with females, only 15 were successful (45%), while for the 18 sessions with males, 14 were successful (78%), resulting in a χ(2) statistic of 4.96, P = .026. However, given all of the χ(2) tests performed in this section, the results must be interpreted with caution because of multiple testing. Results indicate belief in whether the AK test will work was not significantly related to whether it actually did work. A χ(2) test of the relationship between time perception and correct vial choice showed no significant relationships. The χ(2) statistic for the relationship using the hand dynamometer data was 0.927, P = .629. CONCLUSION: The data in this study, particularly when seen in the larger context of a review of the literature from the AK field itself by Klinkoski and Leboeuf (1990), which considered 50 papers published between 1981 and 1987 by the International College of Applied Kinesiology, and the survey by Hall, Lewith, Brien, and Little (2008), using standard evaluation criteria [quality assessment tool for studies of diagnostic accuracy included in systematic reviews (QUADAS), Standards for Reporting of Diagnostic Studies (STARD), JADAD, and Consolidated Standards of Reporting Trials (CONSORT)], for research methodology, as well as six prior non-clinical studies by Radin (1984), Quintanar and Hill (1988), Braud (1989), Arnett et al. (1999), Ludtke (2001), and Kendler and Keating (2003), all together suggest the following: The research published by the Applied Kinesiology field itself is not to be relied upon, and in the experimental studies that do meet accepted standards of science, Applied Kinesiology has not demonstrated that it is a useful or reliable diagnostic tool upon which health decisions can be based.


Asunto(s)
Fuerza de la Mano , Quinesiología Aplicada/métodos , Venenos , Adulto , Técnicas y Procedimientos Diagnósticos , Método Doble Ciego , Femenino , Humanos , Hidroxilamina , Masculino , Edición
11.
Exp Clin Endocrinol Diabetes ; 120(4): 210-6, 2012 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-22328114

RESUMEN

OBJECTIVE: We aimed at evaluating the impact of short and prolonged mild manipulations of intracellular nitric oxide (NO) bioavailability on the main features of insulin secretion and whether NO promotes mitochondrial biogenesis in isolated ß-cells. MATERIALS/METHODS: INS-1E ß-cells were exposed to either the intracellular NO donor, hydroxylamine (HA), or the NO synthase inhibitor, L-nitro-arginine-methyl-ester (L-NAME), at concentrations lower than 2.0 mM. Glucose and arginine-induced insulin secretion (GIIS and AIIS) were measured after short (1 h) or prolonged (48 h) exposure to L-NAME 1.0 and 2.0 mM or HA 0.4 and 0.8 mM, lower concentrations were also evaluated for the 1 h effects. Basal insulin secretion (BIS), with either HA or L-NAME added to culture media, and peroxisome proliferators-activated receptor γ coactivator 1α (PGC-1α), nuclear respiratory factor-1 (NRF-1), and mitochondrial DNA transcription factor-A (Tfam) gene expression during chronic HA supplementation were also measured. RESULTS: Neither L-NAME nor HA affected insulin release at glucose 3.3 mM or in cell culture (BIS). Both short and prolonged cell exposure to L-NAME potentiated GIIS though with a flat dose-response curve while HA inhibited GIIS only at the highest concentration. AIIS was prevented by short exposure to L-NAME and potentiated by HA, while it did not respond to prolonged incubations. Prolonged cell exposure to HA had no effect on PGC-1α, NRF-1 or Tfam gene expression. CONCLUSION: In INS1E cells an intact NO synthesis is necessary to limit insulin release in response to acute glucose gradients and to fully respond to arginine while intracellular NO enrichment above the physiologic levels further inhibits GIIS and potentiate AIIS only when excessive. Prolonged NO manipulations do not affect AIIS, BIS or mitochondrial biogenesis.


Asunto(s)
Hidroxilamina/farmacología , Células Secretoras de Insulina/efectos de los fármacos , NG-Nitroarginina Metil Éster/farmacología , Donantes de Óxido Nítrico/farmacología , Óxido Nítrico/metabolismo , Animales , Arginina/farmacología , Células Cultivadas , Relación Dosis-Respuesta a Droga , Glucosa/farmacología , Insulina/metabolismo , Insulina/farmacología , Secreción de Insulina , Células Secretoras de Insulina/metabolismo , Células Secretoras de Insulina/fisiología , Líquido Intracelular/efectos de los fármacos , Líquido Intracelular/metabolismo , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Mitocondrias/fisiología , Ratas , Factores de Tiempo
12.
FEBS J ; 278(18): 3431-46, 2011 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-21787376

RESUMEN

A split-intein consists of two complementary fragments (N-intein and C-intein) that can associate to carry out protein trans-splicing. The Ssp GyrB S11 split-intein is an engineered unconventional split-intein consisting of a 150-amino-acid N-intein and an extremely small six-amino-acid C-intein, which comprises the conserved intein motif G. Here, we show that fusion proteins containing the 150-amino-acid N-intein could be triggered to undergo controllable N-cleavage in vitro when the six-amino-acid C-intein or a derivative thereof was added as a synthetic peptide in trans. More importantly, we discovered, unexpectedly, that the 150-amino-acid N-intein could be induced by strong nucleophiles to undergo N-cleavage in vitro, and in Escherichia coli cells, in the absence of the motif G-containing six-amino-acid C-intein. This finding indicated that the first step of the protein splicing mechanism (acyl shift) could occur in the absence of the entire motif G. Extensive kinetic analyses revealed that both the motif G residues and the Ser+1 residue positively influenced N-cleavage rate constants and yields. The 150-amino-acid N-intein could also tolerate various unrelated sequences appended to its C-terminus without disruption of the N-cleavage function, suggesting that the catalytic pocket of the intein has considerable structural flexibility. Our findings reveal interesting insights into intein structure-function relationships, and demonstrate a new and potentially more useful method of controllable, intein-mediated N-cleavage for protein engineering applications.


Asunto(s)
Proteínas Bacterianas/química , Girasa de ADN/química , Precursores Enzimáticos/química , Inteínas , Fragmentos de Péptidos/química , Dominios y Motivos de Interacción de Proteínas , Empalme de Proteína , Proteínas Bacterianas/genética , Proteínas Bacterianas/aislamiento & purificación , Proteínas Bacterianas/metabolismo , Catálisis , Girasa de ADN/genética , Girasa de ADN/aislamiento & purificación , Girasa de ADN/metabolismo , Ditiotreitol/farmacología , Precursores Enzimáticos/genética , Precursores Enzimáticos/aislamiento & purificación , Precursores Enzimáticos/metabolismo , Estabilidad de Enzimas , Hidroxilamina/farmacología , Cinética , Mesna/farmacología , Proteínas Mutantes/química , Proteínas Mutantes/genética , Proteínas Mutantes/aislamiento & purificación , Proteínas Mutantes/metabolismo , Oligopéptidos/metabolismo , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/aislamiento & purificación , Fragmentos de Péptidos/metabolismo , Ingeniería de Proteínas/métodos , Empalme de Proteína/efectos de los fármacos , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/aislamiento & purificación , Proteínas Recombinantes de Fusión/metabolismo , Sustancias Reductoras/farmacología , Synechocystis/enzimología , Zinc/farmacología
13.
Neurosci Lett ; 485(1): 74-8, 2010 Nov 12.
Artículo en Inglés | MEDLINE | ID: mdl-20813156

RESUMEN

We investigated whether hydrogen sulfide (H(2)S) may be a mediator of electro-acupuncture (EA) stimulation treatment for hypoxic-ischemic brain-damage (HIBD). We studied a HIBD 7-day-old rat model with 4 types of treatments: (1) 14 sessions of EA; (2) hydroxylamine (HA), an inhibitor of cystathionine-ß-synthase (CBS), the key enzyme of H(2)S generation; (3) both EA and HA; or (4) no treatment. Sham-treated rats with or without EA were also studied. Regional cerebral blood flow (rCBF) was monitored before, during and after EA at different periods of treatment (d1, 7 and 14 sessions). We evaluated motor function, H(2)S levels and CBS expression in the cerebral cortex and prepared cerebral pathomorphological images after 14 sessions of treatment. EA stimulation could increase local blood circulation and improve motor function in HIBD rats. HIBD significantly increased H(2)S levels of brain tissue as compared with sham treatment, and EA treatment could decrease the H(2)S generation. Rats with HIBD receiving both EA and HA therapy showed greatly recovered motor function and brain morphology. H(2)S might be a mediator of EA treatment of HIBD in rats.


Asunto(s)
Electroacupuntura , Sulfuro de Hidrógeno/metabolismo , Hipoxia-Isquemia Encefálica/prevención & control , Animales , Animales Recién Nacidos , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/metabolismo , Corteza Cerebral/patología , Circulación Cerebrovascular , Terapia Combinada , Cistationina betasintasa/antagonistas & inhibidores , Hidroxilamina/uso terapéutico , Hipoxia-Isquemia Encefálica/metabolismo , Hipoxia-Isquemia Encefálica/fisiopatología , Ratas , Ratas Sprague-Dawley
14.
J Oleo Sci ; 59(1): 15-9, 2010.
Artículo en Inglés | MEDLINE | ID: mdl-20032595

RESUMEN

In this study, fatty haydroxamic acids (FHAs), which have biological activities as antibiotics and antifungal, have been synthesized via refluxing of triacylglycrides, palm olein, palm stearin or corn oil with hydroxylamine hydrochloride. The products were characterized using the complex formation test of hydroxamic acid group with zinc(I), copper(II) and iron(III), various technique methods including nuclear magnetic resonance ((1)H NMR) spectroscopy, Fourier transform infrared (FTIR) spectroscopy and elemental analysis. Parameters that may affect the conversion of oils to FHAs including the effect of reaction time, effect of organic solvent and effect of hydro/oil molar issue were also investigated in this study. Results of characterization indicate that FHAs were successfully produced from triacylglycrides. The conversion percentages of palm stearin, palm olein and corn oil into their fatty hydroxamic acids are 82, 81 and 78, respectively. Results also showed that hexane is the best organic solvent to produce the FHAs from the three oils used in this study. The optimum reaction time to achieve the maximum conversion percentage of the oils to FHAs was found to be 10 hours for all the three oils, while the optimum molar ration of hydro/to oil was found to be 7:1 for all the different three oils.


Asunto(s)
Ácidos Grasos/química , Ácidos Grasos/síntesis química , Ácidos Hidroxámicos/química , Ácidos Hidroxámicos/síntesis química , Triglicéridos/química , Antibacterianos/síntesis química , Antibacterianos/química , Antifúngicos/síntesis química , Antifúngicos/química , Ácidos Grasos/metabolismo , Ácidos Hidroxámicos/metabolismo , Hidroxilamina/química , Aceite de Palma , Aceites de Plantas/química , Espectroscopía Infrarroja por Transformada de Fourier
15.
Rapid Commun Mass Spectrom ; 22(12): 1981-92, 2008 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-18494003

RESUMEN

A rapid, simple electrospray ionization mass spectrometry (ESI-MS) sensitivity enhancement method for detecting carbonyl groups in triterpenoids has been developed by using hydroxylamine hydrochloride (NH(2)OH . HCl) as a derivatization reagent. We use the oxime formed during the derivatization reactions and its Beckmann rearrangement intermediates as a means of detecting the carbonyl groups originally present in these triterpenoids. In comparison with other derivatization methods reported in the literature, this method is simple, specific and can be used to detect carbonyl groups in triterpenoids which have low polarity and are poorly or non-ionizable. Moreover, it can also be used to detect hydroxyl groups by using the Dess-Martin periodinane (DMP) to convert primary and secondary hydroxyls into carbonyl groups.


Asunto(s)
Hidroxilamina/química , Espectrometría de Masa por Ionización de Electrospray/métodos , Terpenos/química , Estructura Molecular , Ácido Oleanólico/química , Preparaciones de Plantas/química , Sensibilidad y Especificidad , Factores de Tiempo
16.
Biomacromolecules ; 6(6): 2946-53, 2005.
Artículo en Inglés | MEDLINE | ID: mdl-16283713

RESUMEN

Iron overload is a severe clinical condition and can be largely prevented by the use of iron-specific chelating agents. A successful iron chelator needs to be orally active, nontoxic, and selective. In this study, hydrogels containing pendant hydroxamic acid groups have been synthesized as potential nonabsorbed chelators for iron in the gastrointestinal tract. The synthetic method employed to introduce hydroxamic acid groups to polymer chains involved reaction of polymer gels based on N-acryloxysuccinimide, acryloyl chloride, and (2-hydroxyethyl)acrylate monomers with hydroxylamine. These hydroxamic acid-functionalized polymer gels swell favorably in water and effectively sequester iron. In vitro iron-binding properties of these hydrogels were evaluated from their binding isotherms by use of iron(II) alone and in the presence of other competing metal ions. These polymers bind iron over a broad pH range. The iron-binding properties of the polymers were found to depend on the concentration of hydroxamate groups on polymer chains. The in vivo iron-binding efficacy of the polymers was evaluated in rat as the animal model. The polymers prevented an increase in serum hemoglobin and hematocrit levels in the animals, thus suggesting the prevention of systemic absorption of dietary iron from the gastrointestinal tract. The animals also maintained normal body weight during the treatment period, indicating the absence of any apparent toxicity associated with these polymers.


Asunto(s)
Hidrogeles/química , Ácidos Hidroxámicos/química , Quelantes del Hierro/química , Hierro/química , Polímeros/toxicidad , Acrilamidas/química , Acrilatos/química , Animales , Quelantes/farmacología , Terapia por Quelación/métodos , Ésteres , Tracto Gastrointestinal/metabolismo , Hematócrito , Hemoglobinas/química , Concentración de Iones de Hidrógeno , Hidroxilamina/química , Iones , Hierro de la Dieta/metabolismo , Ligandos , Masculino , Metales , Modelos Químicos , Polímeros/química , Unión Proteica , Ratas , Ratas Wistar , Estireno/química , Succinimidas/química , Factores de Tiempo
17.
Arzneimittelforschung ; 55(1): 30-7, 2005.
Artículo en Inglés | MEDLINE | ID: mdl-15727162

RESUMEN

Two drugs composed of several different plant extracts are in use in Ayurvedic medicine for the treatment of asthma and arthritis, respectively. There is increasing evidence that reactive oxygen species (ROS) arising from several enzymatic reactions are mediators of inflammatory events such as the above mentioned. Plant extracts have the potential for scavenging such reactive oxygen species, dependent on the individual test system. Using biochemical model reactions relevant for the formation of ROS in vivo at inflammatory sites, inhibition of the indicator reaction for the formation of ROS is thought to represent a potential mechanism of the physiological activity of the corresponding preparation.


Asunto(s)
Antioxidantes/química , Medicina Ayurvédica , Antioxidantes/normas , Supervivencia Celular/efectos de los fármacos , Espacio Extracelular/metabolismo , Granulocitos/metabolismo , Humanos , Ácido Hialurónico/metabolismo , Peróxido de Hidrógeno/metabolismo , Radical Hidroxilo/metabolismo , Hidroxilamina/metabolismo , Hipoxantina/metabolismo , Técnicas In Vitro , India , Leucocitos/efectos de los fármacos , Leucocitos/metabolismo , Oxidantes/metabolismo , Peroxidasa/metabolismo , Fenoles/metabolismo , Especies Reactivas de Oxígeno , Superóxidos/metabolismo , Ácido Úrico/metabolismo , Xantina Oxidasa/metabolismo
19.
J Agric Food Chem ; 51(21): 6362-6, 2003 Oct 08.
Artículo en Inglés | MEDLINE | ID: mdl-14518968

RESUMEN

Solutions of 100 mL of 1% commercial pectin each with a different degree of esterification (DE), DE94, DE65, and DE25, were reacted with 100 mL of 2 M alkaline hydroxylamine (pH 12.0) at room temperature for 4 or 18 h. These pectin hydroxamic acids (PHAs; DE94T4, DE94T18, DE65T4, and DE25T4) were used to test the inhibitory activities against semicarbazide-sensitive amine oxidase (SSAO) and angiotensin-converting enzyme (ACE). Compared to different DE pectins (DE94, DE65, and DE25), the PHAs of DE94T4, DE94T18, DE65T4, and DE25T4 showed different inhibition activities against SSAO or ACE. Commercial pectins with different DE values showed negligible SSAO or ACE inhibitions. The order of SSAO inhibition was DE65T4 > DE94T18 approximately DE25T4 >> DE94T4. However, the order of ACE inhibition was DE94T4 > DE94T18 >> DE65T4 > DE25T4. The SSAO activity staining or ACE-hydrolyzed products on TLC chromatogram also confirmed the inhibitory activities of PHAs against SSAO or ACE.


Asunto(s)
Amina Oxidasa (conteniendo Cobre)/antagonistas & inhibidores , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Inhibidores Enzimáticos/farmacología , Ácidos Hidroxámicos/farmacología , Pectinas/química , Pectinas/farmacología , Cromatografía en Capa Delgada , Esterificación , Concentración de Iones de Hidrógeno , Ácidos Hidroxámicos/química , Hidroxilamina/química , Espectrometría de Fluorescencia , Espectrofotometría
20.
Carbohydr Res ; 333(1): 47-58, 2001 Jun 22.
Artículo en Inglés | MEDLINE | ID: mdl-11423110

RESUMEN

A specific, chemical degradation of the methyl esterified galacturonic acid residues of pectins is described. These residues are converted, with hydroxylamine, to hydroxamic acids, and then, with a carbodiimide, to isoureas; the latter undergo a Lossen rearrangement on alkaline hydrolysis. The isocyanates formed are hydrolysed to 5-aminoarabinopyranose derivatives, which spontaneously ring open to give 1,5-dialdehydes. The latter are reduced, in situ, to avoid peeling reactions, with sodium borohydride to give substituted arabitol residues. Thus, overall, partially esterified pectins are specifically cleaved to generate a series of oligogalacturonic acids bearing an arabitol residue as aglycone. Analysis of oligomers so generated discloses the pattern of contiguous nonesterification in a variety of pectins of differing degrees of esterification. Other potential applications are described.


Asunto(s)
Pectinas/química , Esterificación , Etildimetilaminopropil Carbodiimida , Ácidos Hexurónicos/química , Ácidos Hidroxámicos/química , Hidroxilamina , Espectroscopía de Resonancia Magnética , Oxidación-Reducción , Espectrometría de Masa por Ionización de Electrospray
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