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1.
Artículo en Inglés | MEDLINE | ID: mdl-38432457

RESUMEN

Male Japanese quails (Coturnix japonica) have been found to exhibit a three-phase metabolic change when subjected to prolonged fasting, during which basal thermogenesis is significantly reduced. A study had shown that there is a significant difference in the body temperature between male and female Japanese quails. However, whether female Japanese quails also show the same characteristic three-phase metabolic change during prolonged fasting and the underlying thermogenesis mechanisms associated with such changes are still unclear. In this study, female Japanese quails were subjected to prolonged starvation, and the body mass, basal metabolic rate (BMR), body temperature, mass of tissues and organs, body fat content, the state-4 respiration (S4R) and cytochrome c oxidase (CCO) activity in the muscle and liver of these birds were measured to determine the status of metabolic changes triggered by the starvation. In addition, the levels of glucose, triglyceride (TG) and uric acid, and thyroid hormones (T3 and T4) in the serum and the mRNA levels of myostatin (MSTN) and avian uncoupling protein (av-UCP) in the muscle were also measured. The results revealed the existence of a three-phase stage similar to that found in male Japanese quails undergoing prolonged starvation. Fasting resulted in significantly lower body mass, BMR, body temperature, tissues masses and most organs masses, as well as S4R and CCO activity in the muscle and liver. The mRNA level of av-UCP decreased during fasting, while that of MSTN increased but only during Phase I and II and decreased significantly during Phase III. Fasting also significantly lowered the T3 level and the ratio of T3/T4 in the serum. These results indicated that female Japanese quails showed an adaptive response in basal thermogenesis at multiple hierarchical levels, from organismal to biochemical, enzyme and cellular level, gene and endocrine levels and this integrated adjustment could be a part of the adaptation used by female quails to survive long-term fasting.


Asunto(s)
Coturnix , Codorniz , Femenino , Masculino , Animales , Coturnix/metabolismo , Codorniz/metabolismo , Ayuno/metabolismo , Termogénesis , ARN Mensajero/genética
2.
Chem Biodivers ; 21(6): e202400524, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38634793

RESUMEN

Baihe is a commonly used Chinese medicine for the treatment of neurological disorders. Clinically, the bulbs of Lilium brownii are used to act as Baihe. In the study, two new phenylpropanoid compounds including 3-O-acetyl-1-O-caffeoylglycerol (1) and 3-O-acetyl-1-O-p-coumaroylglycerol (2) were isolated from the bulbs of L. brownii. Their structures were identified by spectroscopic method and the effect on monoamine oxidase activity was determined using an enzyme labeling method. The results show 1 and 2 have anti-monoamine oxidase activity with 20.96 % and 22.31 % inhibition rates at 50 µg/ml, respectively.


Asunto(s)
Lilium , Inhibidores de la Monoaminooxidasa , Monoaminooxidasa , Lilium/química , Monoaminooxidasa/metabolismo , Inhibidores de la Monoaminooxidasa/farmacología , Inhibidores de la Monoaminooxidasa/química , Inhibidores de la Monoaminooxidasa/aislamiento & purificación , Estructura Molecular , Raíces de Plantas/química , Relación Estructura-Actividad , Relación Dosis-Respuesta a Droga
3.
Molecules ; 29(6)2024 Mar 07.
Artículo en Inglés | MEDLINE | ID: mdl-38542837

RESUMEN

Nonsymbiotic phytoglobins (nsHbs) are a diverse superfamily of hemoproteins grouped into three different classes (1, 2, and 3) based on their sequences. Class 1 Hb are expressed under hypoxia, osmotic stress, and/or nitric oxide exposure, while class 2 Hb are induced by cold stress and cytokinins. Both are mainly six-coordinated. The deoxygenated forms of the class 1 and 2 nsHbs from A. thaliana (AtHb1 and AtHb2) are able to reduce nitrite to nitric oxide via a mechanism analogous to other known globins. NsHbs provide a viable pH-dependent pathway for NO generation during severe hypoxia via nitrite reductase-like activity with higher rate constants compared to mammalian globins. These high kinetic parameters, along with the relatively high concentrations of nitrite present during hypoxia, suggest that plant hemoglobins could indeed serve as anaerobic nitrite reductases in vivo. The third class of nsHb, also known as truncated hemoglobins, have a compact 2/2 structure and are pentacoordinated, and their exact physiological role remains mostly unknown. To date, no reports are available on the nitrite reductase activity of the truncated AtHb3. In the present work, three representative nsHbs of the plant model Arabidopsis thaliana are presented, and their nitrite reductase-like activity and involvement in nitrosative stress is discussed. The reaction kinetics and mechanism of nitrite reduction by nsHbs (deoxy and oxy form) at different pHs were studied by means of UV-Vis spectrophotometry, along with EPR spectroscopy. The reduction of nitrite requires an electron supply, and it is favored in acidic conditions. This reaction is critically affected by molecular oxygen, since oxyAtHb will catalyze nitric oxide deoxygenation. The process displays unique autocatalytic kinetics with metAtHb and nitrate as end-products for AtHb1 and AtHb2 but not for the truncated one, in contrast with mammalian globins.


Asunto(s)
Arabidopsis , Nitritos , Animales , Nitritos/química , Óxido Nítrico/metabolismo , Hemoglobinas/química , Nitrito Reductasas/química , Hipoxia , Arabidopsis/metabolismo , Oxidación-Reducción , Mamíferos/metabolismo
4.
Saudi Pharm J ; 32(2): 101936, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38261938

RESUMEN

In this work, we investigated Diospyros kaki extract and an isolated compound for their potential as xanthine oxidase (XO) inhibitors, a target enzyme involved in inflammatory disorders. The prepared extract was subjected to column chromatography, and dinaphthodiospyrol S was isolated. Then XO inhibitory properties were assessed using a spectrophotometry microplate reader. DMSO was taken as a negative control, and allopurinol was used as a standard drug. The molecular docking study of the isolated compound to the XO active site was performed, followed by visualization and protein-ligand interaction. The defatted chloroform extract showed the highest inhibitory effect, followed by the chloroform extract and the isolated compound. The isolated compound exhibited significant inhibitory activity against XO with an IC50 value of 1.09 µM. Molecular docking studies showed that the compound strongly interacts with XO, forming hydrogen bond interactions with Arg149 and Cys113 and H-pi interactions with Cys116 and Leu147. The binding score of -7.678 kcal/mol further supported the potential of the isolated compound as an XO inhibitor. The quantum chemical procedures were used to study the electronic behavior of dinaphthodiospyrol S isolated from D. kaki. Frontier molecular orbital (FMO) analysis was performed to understand the distribution of electronic density, highest occupied molecular orbital HOMO, lowest unoccupied molecular orbital LUMO, and energy gaps. The values of HOMO, LUMO, and energy gap were found to be -6.39, -3.51 and 2.88 eV respectively. The FMO results indicated the intramolecular charge transfer. Moreover, reactivity descriptors were also determined to confirm the stability of the compound. The molecular electrostatic potential (MEP) investigation was done to analyze the electrophilic and nucleophilic sites within a molecule. The oxygen atoms in the compound exhibited negative potential, indicating that they are favorable sites for electrophilic attacks. The results indicate its potential as a therapeutic agent for related disorders. Further studies are needed to investigate this compound's in vivo efficacy and safety as a potential drug candidate.

5.
Clin Immunol ; 251: 109316, 2023 06.
Artículo en Inglés | MEDLINE | ID: mdl-37055004

RESUMEN

Chronic granulomatous disease (CGD) is a human IEI caused by mutations in genes encoding the NADPH oxidase subunits, the enzyme responsible for the respiratory burst. CGD patients have severe life-threatening infections, hyperinflammation and immune dysregulation. Recently, an additional autosomal recessive AR-CGD (type 5) caused by mutations in CYBC1/EROS gene was identified. We report a AR-CGD5 patient with a novel loss of function (LOF) homozygous deletion c.8_7del in the CYBC1 gene including the initiation ATG codon that leads to failure of CYBC1/EROS protein expression and presenting with an unusual clinical manifestation of childhood-onset sarcoidosis-like disease requiring multiple immunosuppressive therapies. We described an abnormal gp91phox protein expression/function in the patient's neutrophils and monocytes (about 50%) and a severely compromised B cell subset (gp91phox < 15%; DHR+ < 4%). Our case-report emphasized the importance of considering a diagnosis of AR-CGD5 deficiency even in absence of typical clinical and laboratory findings.


Asunto(s)
Enfermedad Granulomatosa Crónica , Humanos , Femenino , Enfermedad Granulomatosa Crónica/genética , Enfermedad Granulomatosa Crónica/diagnóstico , Homocigoto , Eliminación de Secuencia/genética , NADPH Oxidasas/genética , Mutación , Fenotipo
6.
Mikrochim Acta ; 190(5): 196, 2023 04 29.
Artículo en Inglés | MEDLINE | ID: mdl-37118111

RESUMEN

Nitrated α-syn (nitro-α-syn) is a biomarker for Parkinson's desease (PD), and its sensitive detection in serum is of great importance for early PD diagnosis. Silver-coated copper MOF (Cu-MOF@Ag) with outstanding oxidase activity and electrochemical response property was designed and synthesized. Cu-MOF@Ag exhibited excellent oxidase activity with a low Km value (0.568 mM), avoiding the addition of strong oxidant to catalyze chromogenic substrate, which enhanced the colorimetric stability. Silver nanoparticles as an electrochemical signal reporter can be easily decorated on the surface of Cu-MOF with bifunctional groups (-SH and -NH2) material, which can increase the electrochemical signal output. The α-syn antibody modified Cu-MOF@Ag and nitro-α-syn modified magnetic nanoparticle were used as immunoprobes to specifically capture nitro-α-syn. A dual-modal immunosensor was fabricated for the simple and reliable detection of nitro-α-syn based on Cu-MOF@Ag. Combing colorimetric and electrochemical detection, nitro-α-syn can be determined quantitatively within a wide linear range (10-350 ng/mL) with low detection limit (0.5 ng/mL). The ability of the sensor with magnetic separation and dual signal analysis allowed to successfully detect nitro-α-syn and distinguish PD patients from healthy people (P < 0.005). Thanks to its excellent selectivity, stability, and the precision of 2.69%, the dual-modal sensor has potential clinical application for nitro-α-syn detection and paves a new way for PD diagnosis at its early stage.


Asunto(s)
Técnicas Biosensibles , Nanopartículas del Metal , Estructuras Metalorgánicas , Humanos , alfa-Sinucleína , Nitratos , Plata/química , Estructuras Metalorgánicas/química , Nanopartículas del Metal/química , Inmunoensayo , Oxidorreductasas
7.
Molecules ; 28(13)2023 Jun 27.
Artículo en Inglés | MEDLINE | ID: mdl-37446700

RESUMEN

This paper presents the effects of irrigating barley plants with different type of water solutions saturated with gaseous ozone generated from atmospheric air. The study investigated the effects of the applied types of water on the modulation of the biosynthesis of selected bioactive compounds (content of total polyphenols, small molecule antioxidants, vitamin C) in the produced plant material. A number of transformations of reactive oxygen species (ROS) and nitrogen compounds have also been postulated; these are observed during the saturation of water with gaseous O3 and 30 min after the end of the process. It was shown that after the process of water saturation with gaseous O3, the gas later is converted to compounds with high oxidative potential and good stability; these, in turn, lead to the oxidation of oxidates generated from atmospheric nitrogen into nitrates, which exhibit fertilising properties. Thirty minutes after the process of H2O saturation with gaseous O3 was completed, the tests showed the highest concentrations of nitrates and the relatively high oxidative potential of the solution originating from H2O2 with a low concentration of the dissolved O3. This solution exhibited the highest activity modulating the biosynthesis of polyphenols, small molecule antioxidants and vitamin C in young barley plants. The resulting differences were significant, and they were reflected by 15% higher total polyphenol content, 35% higher antioxidative potential and 57% greater content of vitamin C compared to the control specimens (plants treated with fresh H2O).


Asunto(s)
Hordeum , Ozono , Antioxidantes/farmacología , Agua , Peróxido de Hidrógeno , Nitratos , Ácido Ascórbico , Ozono/farmacología , Polifenoles , Plantas
8.
Support Care Cancer ; 30(11): 9369-9377, 2022 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-36112225

RESUMEN

PURPOSE: The relationship between activity of the small intestinal villi and the effectiveness of chemotherapy remains unclear. This study aimed to investigate how serum diamine oxidase (DAO) activity affects antitumor effects, adverse events, and amino acid absorption. METHODS: We performed a single-center prospective cohort study that enrolled 50 patients with esophageal cancer (EC) receiving docetaxel, cisplatin, and 5-fluorouracil therapy. We determined the cut-off value of serum DAO activity contributing to a response to chemotherapy using a generalized additive model. Additionally, we compared adverse events, inflammatory markers, blood amino acid levels, and quality of life between the high and low DAO activity groups during chemotherapy. RESULTS: The cut-off value of serum DAO activity at the first visit that contributed to a chemotherapy response was 6.5 units/L. Leukopenia and neutropenia of grade ≥ 3 were significantly higher in the DAO low (< 6.5 units/L) group (p = 0.044, 0.017, respectively). Interleukin-6 was significantly lower in the DAO high (≥ 6.5 units/L) group at the first visit and at 4 weeks after the end of chemotherapy (p = 0.039, 0.011, respectively). Glutamine was higher in the DAO high group at all measurement points during chemotherapy. Fatigue was significantly lower in the DAO high group (p = 0.001). CONCLUSION: Serum DAO activity may be a predictor of the response to chemotherapy in patients with EC. The absorption capacity of amino acids was maintained in the group with high DAO activity, which may have contributed to the anti-inflammatory effect and provided a background for reducing adverse events.


Asunto(s)
Amina Oxidasa (conteniendo Cobre) , Antineoplásicos , Neoplasias , Humanos , Amina Oxidasa (conteniendo Cobre)/metabolismo , Glutamina , Mucosa Intestinal/patología , Estudios Prospectivos , Calidad de Vida , Neoplasias/tratamiento farmacológico , Antineoplásicos/efectos adversos , Antineoplásicos/uso terapéutico
9.
Eur Heart J ; 42(48): 4947-4960, 2021 12 21.
Artículo en Inglés | MEDLINE | ID: mdl-34293101

RESUMEN

AIMS: Recent clinical trials indicate that sodium-glucose cotransporter 2 (SGLT2) inhibitors improve cardiovascular outcomes in heart failure patients, but the underlying mechanisms remain unknown. We explored the direct effects of canagliflozin, an SGLT2 inhibitor with mild SGLT1 inhibitory effects, on myocardial redox signalling in humans. METHODS AND RESULTS: Study 1 included 364 patients undergoing cardiac surgery. Right atrial appendage biopsies were harvested to quantify superoxide (O2.-) sources and the expression of inflammation, fibrosis, and myocardial stretch genes. In Study 2, atrial tissue from 51 patients was used ex vivo to study the direct effects of canagliflozin on NADPH oxidase activity and nitric oxide synthase (NOS) uncoupling. Differentiated H9C2 and primary human cardiomyocytes (hCM) were used to further characterize the underlying mechanisms (Study 3). SGLT1 was abundantly expressed in human atrial tissue and hCM, contrary to SGLT2. Myocardial SGLT1 expression was positively associated with O2.- production and pro-fibrotic, pro-inflammatory, and wall stretch gene expression. Canagliflozin reduced NADPH oxidase activity via AMP kinase (AMPK)/Rac1signalling and improved NOS coupling via increased tetrahydrobiopterin bioavailability ex vivo and in vitro. These were attenuated by knocking down SGLT1 in hCM. Canagliflozin had striking ex vivo transcriptomic effects on myocardial redox signalling, suppressing apoptotic and inflammatory pathways in hCM. CONCLUSIONS: We demonstrate for the first time that canagliflozin suppresses myocardial NADPH oxidase activity and improves NOS coupling via SGLT1/AMPK/Rac1 signalling, leading to global anti-inflammatory and anti-apoptotic effects in the human myocardium. These findings reveal a novel mechanism contributing to the beneficial cardiac effects of canagliflozin.


Asunto(s)
Canagliflozina , Inhibidores del Cotransportador de Sodio-Glucosa 2 , Canagliflozina/metabolismo , Canagliflozina/farmacología , Humanos , Miocardio , Miocitos Cardíacos/metabolismo , Oxidación-Reducción , Inhibidores del Cotransportador de Sodio-Glucosa 2/farmacología
10.
Microb Pathog ; 153: 104637, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33232765

RESUMEN

Ten marine species, including different crabs, bivalve molluscs, and fish intestines were selected to screen the natural antimicrobial protein or peptide as they are enriched with various microorganisms. The crude extract from Varuna litterata, a marine crab which is used as a raw material in the preparation of pickled crabs in Chaoshan area of China, was proved to have a potent bacteriostatic effect against gram-negative bacterium (Escherichia coli) and gram-positive bacterium(Staphylococcus aureus) compared with other marine species. The crude proteins of Varunalitterata were salted-out for preliminary purification and further purified by gel filtration (Sephadex G-150) or anion exchange (DEAE-cellulose 52) chromatographic column. An increase in the antimicrobial activity was noted with the increase in the purity level of the protein. A relatively pure protein was eventually obtained, which was determined to be belonging to the hemocyanin family based on the mass spectrometric data analysis. The purified proteins solution (1 mg/ml) from Varuna litterata exhibited similar antimicrobial activity to that of gentamycin sulfate (0.2 mg/ml), which were relatively stable in a certain pH or temperature range. A structure-activity relationship of the purified hemocyanin was determined based on the interaction of hemocyanin and different chromatographic medium, which revealed that the integrated hexamers played a remarkable role in its bacteriostatic activity. Moreover, the phenoloxidase activity of hemocyanin from Varuna litterata was found as the underlying cause of its antimicrobial potential.


Asunto(s)
Antiinfecciosos , Braquiuros , Animales , Antibacterianos/farmacología , Antiinfecciosos/farmacología , China , Staphylococcus aureus
11.
Arch Microbiol ; 203(7): 4677-4692, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-34180014

RESUMEN

The problem of arsenic (As) pollution being severe warrants opting for low-cost microbial remediation strategies. The present study of identifying suitable bacterial strains led to the isolation of eleven As-tolerant strains from the As-contaminated rhizosphere soils of West Bengal, India. They were found to oxidize/reduce 55-31.6% of 5 mM As(III) and 73-37.6% of 5 mM As(V) within 12 h. The four isolates (BcAl-1, JN 73, LAR-2, and AR-30) had a high level of As(III) oxidase activity along with a higher level of As(V) and As(III) resistance. The agar diffusion assay of the isolates further confirmed their ability to endure As stress. The presence of aoxB gene was observed in these four As(III) oxidizing isolates. Evaluation of plant growth-promoting characteristics revealed that BcAl-1 (Burkholderia cepacia), JN 73 (Burkholderia metallica), AR-30 (Burkholderia cenocepacia), and LAR-2 (Burkholderia sp.) had significant plant growth-promoting characteristics (PGP), including the ability to solubilize phosphate, siderophore production, indole acetic acid-like molecules production, ACC deaminase production, and nodule formation under As stressed condition. BcAl-1 and JN 73 emerged as the most promising traits in As removal as well as plant growth promotion.


Asunto(s)
Burkholderia , Plantas , Microbiología del Suelo , Arsénico/metabolismo , Arsenitos/metabolismo , Burkholderia/fisiología , India , Oxidación-Reducción , Plantas/microbiología , Suelo/química , Contaminantes del Suelo/metabolismo
12.
Sensors (Basel) ; 21(22)2021 Nov 14.
Artículo en Inglés | MEDLINE | ID: mdl-34833643

RESUMEN

In recent years, cerium oxide (CeO2) nanoparticles (NPs) have drawn significant attention owing to their intrinsic enzyme mimetic properties, which make them powerful tools for biomolecular detection. In this work, we evaluated the effect of pyrophosphate (PPi) on the oxidase activity of CeO2 NPs. The presence of PPi was found to enhance the oxidase activity of CeO2 NPs, with enhanced colorimetric signals. This particular effect was then used for the colorimetric detection of target nucleic acids. Overall, the PPi-enhanced colorimetric signals of CeO2 NPs oxidase activity were suppressed by the presence of the target nucleic acids. Compared with previous studies using CeO2 NPs only, our proposed system significantly improved the signal change (ca. 200%), leading to more sensitive and reproducible colorimetric analysis of target nucleic acids. As a proof-of-concept study, the proposed system was successfully applied to the highly selective and sensitive detection of polymerase chain reaction products derived from Klebsiella pneumoniae. Our findings will benefit the rapid detection of nucleic acid biomarkers (e.g., pathogenic bacterial DNA or RNA) in point-of-care settings.


Asunto(s)
Cerio , Nanopartículas , Ácidos Nucleicos , Colorimetría , Difosfatos , Oxidorreductasas
13.
Amino Acids ; 52(8): 1089-1105, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32844248

RESUMEN

The versatility of reactions catalyzed by pyridoxal 5'-phosphate (PLP) enzymes is largely due to the chemistry of their extraordinary catalyst. PLP is necessary for many reactions involving amino acids. Reaction specificity is controlled by the orientation of the external aldimine intermediate that is formed upon addition of the amino acidic substrate to the coenzyme. The breakage of a specific bond of the external aldimine gives rise to a carbanionic intermediate. From this point, the different reaction pathways diverge leading to multiple activities: transamination, decarboxylation, racemization, elimination, and synthesis. A significant novelty appeared approximately 30 years ago when it was reported that some PLP-dependent decarboxylases are able to consume molecular oxygen transforming an amino acid into a carbonyl compound. These side paracatalytic reactions could be particularly relevant for human health, also considering that some of these enzymes are responsible for the synthesis of important neurotransmitters such as γ-aminobutyric acid, dopamine, and serotonin, whose dysregulation under oxidative conditions could have important implications in neurodegenerative states. However, the reactivity of PLP enzymes with dioxygen is not confined to mammals/animals. In fact, some plant PLP decarboxylases have been reported to catalyze oxidative reactions producing carbonyl compounds. Moreover, other recent reports revealed the existence of new oxidase activities catalyzed by new PLP enzymes, MppP, RohP, Ind4, CcbF, PvdN, Cap15, and CuaB. These PLP enzymes belong to the bacterial and fungal kingdoms and are present in organisms synthesizing bioactive compounds. These new PLP activities are not paracatalytic and could only scratch the surface on a wider and unexpected catalytic capability of PLP enzymes.


Asunto(s)
Aminoácidos/biosíntesis , Carboxiliasas/química , Carboxiliasas/metabolismo , Fosfato de Piridoxal/química , Aminoácidos/química , Animales , Bacterias/enzimología , Biocatálisis , Dopa-Decarboxilasa/metabolismo , Hongos/enzimología , Humanos , Oxidación-Reducción , Oxígeno/metabolismo , Plantas/enzimología
14.
Mikrochim Acta ; 187(8): 465, 2020 07 20.
Artículo en Inglés | MEDLINE | ID: mdl-32691158

RESUMEN

Cu2+ are found to greatly reduce the photoinduced oxidase activity of fluorescein and then inhibit the chromogenic reaction catalyzed by fluorescein. A simple colorimetric assay for Cu2+ is established. Based on this, bifunctional nanocomposites of α-fetoprotein (AFP) antibody (Ab) and copper-based metal-organic framework (Ab2@Cu-MOF) are synthesized by the simple self-assembly of AFP Ab2, Cu2+, and 4,4'-dipyridyl: the binding site of AFP Ab2 exposed on the surface of the nanocomposites can specifically recognize AFP antigen; Cu2+ in nanocomposites can inhibit the visible light-induced activity of fluorescein. The structure of Ab2@Cu-MOF disintegrate and Cu2+ is released in an acetate buffer solution. The higher the amount of AFP antigens, the more significant the inhibitory effect. Thus, the Ab2@Cu-MOF immunoassay for AFP determination is established using 3,3',5,5'-tetramethylbenzidine as chromogenic substrate with a detection limit of 35 pg.mL-1. This simple, cheap, and sensitive method sheds substantial light on practical clinical diagnosis. Meanwhile, the mechanism of inhibition is revealed to facilitate the targeted selection of enzyme regulators. Graphical abstract Diagrammatic illustration of Cu2+ detection (part a) and Ab2@Cu-MOF immunoassay for sensing α-fetoprotein based on the synthesized Ab2@Cu-MOF nanocomposites (parts a and b).


Asunto(s)
Anticuerpos Biespecíficos/inmunología , Cobre/química , Estructuras Metalorgánicas/química , Nanocompuestos/química , alfa-Fetoproteínas/análisis , Anticuerpos Inmovilizados/inmunología , Bencidinas/química , Compuestos Cromogénicos/química , Colorimetría , Fluoresceína/química , Colorantes Fluorescentes/química , Humanos , Inmunoensayo/métodos , Límite de Detección , Piridinas/química , alfa-Fetoproteínas/inmunología
15.
Angew Chem Int Ed Engl ; 59(47): 20940-20945, 2020 11 16.
Artículo en Inglés | MEDLINE | ID: mdl-32701181

RESUMEN

Tyrosinases (TYRs) catalyze the hydroxylation of phenols and the oxidation of the resulting o-diphenols to o-quinones, while catechol oxidases (COs) exhibit only the latter activity. Aurone synthase (AUS) is not able to react with classical tyrosinase substrates, such as tyramine and l-tyrosine, while it can hydroxylate its natural substrate isoliquiritigenin. The structural difference of TYRs, COs, and AUS at the heart of their divergent catalytic activities is still a puzzle. Therefore, a library of 39 mutants of AUS from Coreopsis grandiflora (CgAUS) was generated and the activity studies showed that the reactivity of the three conserved histidines (HisA2 , HisB1 , and HisB2 ) is tuned by their adjacent residues (HisB1 +1, HisB2 +1, and waterkeeper residue) either to react as stronger bases or / and to stabilize a position permissive for substrate proton shuffling. This provides the understanding for C-H activation based on the type-III copper center to be used in future biotechnological processes.


Asunto(s)
Aminoácidos/análisis , Catecol Oxidasa/metabolismo , Cobre/metabolismo , Monofenol Monooxigenasa/metabolismo , Aminoácidos/metabolismo , Catecol Oxidasa/química , Cobre/química , Coreopsis/enzimología , Modelos Moleculares , Monofenol Monooxigenasa/química
16.
Microbiol Immunol ; 62(9): 567-573, 2018 09.
Artículo en Inglés | MEDLINE | ID: mdl-30118139

RESUMEN

Capnocytophaga canimorsus and Capnocytophaga cynodegmi, both commensal bacteria in the oral cavities of dogs and cats, are zoonotic pathogens. In particular, C. canimorsus causes sepsis and fatal septic shock. Recently, a novel Capnocytophaga species, C. canis, was isolated from the oral cavities of healthy dogs. It is reportedly oxidase-negative and therefore considered avirulent in humans. In the present study, three strains of C. canis were isolated from Japanese patients with sepsis. All three strains, HP20001, HP33001 and HP40001, were oxidase-positive. Nucleotide sequence identities of the 16S rRNA gene of the three strains to the C. canimorsus type strain ATCC35979, C. cynodegmi type strain ATCC49044 and C. canis type strain LMG29146 were 96.9-97.0%, 96.9-97.0% and 99.7-99.8%, respectively. Multi-locus sequence analysis based on seven house-keeping genes, dnaJ, fumC, glyA, gyrB, murG, trpB and tuf, revealed that the oxidase-positive C. canis strains isolated in Japan and oxidase-negative strains of C. canis from canine oral cavities in Switzerland were clustered in different genetic subgroups. These results indicate that the virulence of C. canis strains in humans is associated with oxidase activity.


Asunto(s)
Capnocytophaga/clasificación , Capnocytophaga/aislamiento & purificación , Capnocytophaga/patogenicidad , Infecciones por Bacterias Gramnegativas/microbiología , Filogenia , Sepsis/microbiología , Anciano , Anciano de 80 o más Años , Animales , Técnicas de Tipificación Bacteriana , Composición de Base , Benzoquinonas/análisis , Mordeduras y Picaduras/microbiología , Capnocytophaga/genética , Enfermedades de los Gatos/microbiología , Gatos , Girasa de ADN/genética , ADN Bacteriano/aislamiento & purificación , Enfermedades de los Perros/microbiología , Perros , Femenino , Genes Bacterianos/genética , Humanos , Japón , Masculino , Persona de Mediana Edad , Tipificación de Secuencias Multilocus , Fenotipo , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Especificidad de la Especie , Secuenciación Completa del Genoma , Zoonosis/microbiología
17.
Bull Entomol Res ; 108(4): 494-500, 2018 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-29061198

RESUMEN

Gall-making Cynipidae manipulate the leaves of host plant to form galls where offspring find shelter and food. The relationship between oak gallwasp and biochemical mechanisms of galls still requires a better understanding. So, in this research, protein and phenolic compound contents, as well as the activity of antioxidative enzymes and pathogenesis-related (PR) proteins were determined. Galls caused by asexual generation of Cynips quercusfolii L., Neuroterus numismalis (Fourc.) and N. quercusbaccarum L., as a model were used. All cynipid species modified the protein levels of gall tissues, but they cannot be treated as protein sinks. Significantly higher levels of phenols were observed in the galled leaves and galls of all cynipid species when compared with the control tissues. Peroxidase and polyphenol oxidase activity was usually low or showed no activity in galled tissues of all species. PR proteins, such as chitinase and ß-1,3-glucanase, had a similar activity profile. Their activity significantly increased in the leaves with galls of all cynipid species, especially those infested with C. quercusfolii. Data generated in this study clearly indicate that galling Cynipidae manipulate the biochemical machinery of the galls for their own needs. However, the pattern of the biochemical features of leaves with galls and galled tissues depends on gall-making species.


Asunto(s)
Tumores de Planta/parasitología , Quercus/parasitología , Árboles/parasitología , Avispas/fisiología , Animales , Catecol Oxidasa/metabolismo , Quitinasas/metabolismo , Glucano 1,3-beta-Glucosidasa/metabolismo , Peroxidasa/metabolismo , Hojas de la Planta/metabolismo , Quercus/metabolismo , Reproducción Asexuada/fisiología , Árboles/metabolismo
18.
J Food Sci Technol ; 55(4): 1306-1313, 2018 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-29606744

RESUMEN

The effect of shelf storage under ambient conditions of cut apple dices on degradation of bioactive compounds such ascorbic acid, total phenols, antioxidant activity (% DPPH inhibition) and PPO activity were investigated. The results indicated that antioxidant activity declined significantly over 80 min storage of diced apples at ambient temperature. Similar trend was observed for ascorbic acid, total phenols and PPO activity. Ascorbic acid, total phenols and antioxidant activity degradation followed first-order kinetics where the rate constant (k) was found to be in range for all the thirteen cultivars, though initial ascorbic acid and phenol content varied in different apple cultivars. The reaction rate constant (k) for first order degradation ranged from 1.16 to 1.97, 0.89 to 1.29 and 0.37 to 1.54 for antioxidant activity, total phenols and ascorbic acid, respectively. This explains that antioxidant activity degrades at higher rate than total phenols and ascorbic acid, which also corroborates that antioxidant activity is affected by both total phenols and ascorbic acid content. In general, total antioxidant activity for apple dices kept for 80 min under ambient conditions exhibited lower values as compared to control.

19.
J Food Sci Technol ; 55(8): 2935-2944, 2018 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-30065402

RESUMEN

Being climacteric fruit the organoleptic quality of Guava deteriorates due to continuously occurring biochemical changes during the storage leading to heavy post harvest losses. The present research work was planned to depreciate the losses by assessing the effect of different individual packagings on the shelf life of the fruit. The individual wrapping of fruits were carried out in LDPE films by cling, shrink, vacuum and modified atmosphere packaging (MAP) and stored at room temperature (37 °C). Individual wrapping delayed the ripening process by slowing down the metabolic activities and reducing the magnitude of changes in physicochemical parameters. In control fruits significant compositional changes along with the total phenol content and ascorbic acid were observed with higher decay loss. However, wrapping of fruits maintained the natural freshness and helped in retaining the marketability of the fruits. Vacuum packing and MAP showed minimum PLW (3.5%), decay loss and ripening during storage. Cling wrapping and shrink wrapping were the best treatments that enhanced the shelf life of fruits by 4 days at room temperature.

20.
Chemistry ; 23(6): 1346-1352, 2017 Jan 26.
Artículo en Inglés | MEDLINE | ID: mdl-27726210

RESUMEN

In a natural geochemical cycle, manganese-oxide minerals (MnOx ) are principally formed through a microbial process, where a putative multicopper oxidase MnxG plays an essential role. Recent success in isolating the approximately 230 kDa, enzymatically active MnxEFG protein complex, has advanced our understanding of biogenic MnOx mineralization. Here, the kinetics of MnOx formation catalyzed by MnxEFG are examined using a quartz crystal microbalance (QCM), and the first electrochemical characterization of the MnxEFG complex is reported using Fourier transformed alternating current voltammetry. The voltammetric studies undertaken using near-neutral solutions (pH 7.8) establish the apparent reversible potentials for the Type 2 Cu sites in MnxEFG immobilized on a carboxy-terminated monolayer to be in the range 0.36-0.40 V versus a normal hydrogen electrode. Oxidative priming of the MnxEFG protein complex substantially enhances the enzymatic activity, as found by in situ electrochemical QCM analysis. The biogeochemical significance of this enzyme is clear, although the role of an oxidative priming of catalytic activity might be either an evolutionary advantage or an ancient relic of primordial existence.


Asunto(s)
Compuestos de Manganeso/metabolismo , Óxidos/metabolismo , Oxidorreductasas/metabolismo , Biocatálisis , Técnicas Electroquímicas , Cinética , Microscopía Electrónica de Rastreo , Tecnicas de Microbalanza del Cristal de Cuarzo , Espectrometría por Rayos X
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