ABSTRACT
Abstract The unconscious use of pesticides causes various adverse effects on non-target organisms, including humans. Enzymes that control metabolism become the target of the pesticide and the organs are damaged due to toxic effects. Glutathione s-transferase (GST, EC 2.5.1.18), an important enzyme of the detoxification mechanism and antioxidant defense system, can be affected by such toxic substances. Therefore, the effect of fenarimol on GST enzyme activity was investigated in our study. For this, 200 mg/kg fenarimol was administered intraperitoneally to male and female rats at different periods (2, 4, 8, 16, 32, 64 and 72 hours). After application, GST enzyme activity was analysed in the liver, kidney, brain and small intestine tissues of the rats. According to our results, activation (liver, kidney, small intestine) or inhibition (brain) of the generally GST enzyme was observed in the tissues of rats exposed to fenarimol. It is thought that the increase and/or decrease in this enzyme activity may be the cause of the toxic effect of fenarimol.
Subject(s)
Animals , Rats , Pesticides/adverse effects , Glutathione S-Transferase pi , Enzyme Activation , Fungicides, Industrial/adverse effectsABSTRACT
Abstract The evolution of species is inevitably accompanied by the evolution of metabolic networks to adapt to different environments. The metabolic networks of different species were collected from the Kyoto Encyclopedia of Genes and Genomes (KEGG) website, and some enzyme reactions with the highest occurrence frequency in all species were found and are reported in this paper. The correlation coefficients of whether the enzyme reactions appear in all species were calculated, and the corresponding evolutionary correlation connection networks were calculated according to different correlation coefficient thresholds. These studies show that, as the evolutionary correlation of enzyme reactions increases, the weighted average of the mean functional concentration ratios of the enzyme reactions also increases, indicating that the functional concentration ratio of enzyme reactions has a certain correlation with the evolutionary correlation. The work presented in this paper enhances our understanding of the characteristics and general rules of metabolic network evolution.
Subject(s)
Enzyme Activation , Metabolic Networks and Pathways , Adaptation, Biological , MetabolismABSTRACT
Abstract Ananas Comosus (also known as pineapple) is a part of Bromeliaceae family and it is consumed as food as well as folk medicine for the treatment of various diseases. It is reported that pineapple is a rich source of bromelain, a cysteine protease and it is considered as an important enzyme in different industries due to its significant therapeutic and industrial applications such as anticancer, anti-inflammatory and meat tenderizing. Bromelain is mostly present in fruit and stem of pineapple, but it is reported that crown, core, and peels, which constitute the waste of the pineapple plant, also contain bromelain but limited data is available. Therefore, the proposed study aimed at utilizing pineapple waste for the extraction and characterization of bromelain. Firstly, crude bromelain was extracted with phosphate buffer (pH 7), then it was subjected to partial purification using different fractions of ammonium sulphate (NH4)2SO4 such as 30, 40, 50 and 60% followed by desalting and concentration. Enzyme activity was calculated by using casein digesting unit (CDU) method. The results demonstrated that the crown bromelain showed highest purification of 4.34-fold at 30% (NH4)2SO4 saturation, whereas core and peel bromelain showed highest purification of 2.75 and 2.59-fold at 40% (NH4)2SO4 saturation. The molecular weight of crude and partially purified bromelain was determined by SDS-PAGE analysis and found to be 26 KDa. The pH and thermal stability of all the parts of pineapple showed maximum stability at pH 7 and at 35oC temperature.
Subject(s)
Bromelains/isolation & purification , Enzyme Activation , Ammonium Sulfate , Peptide Hydrolases , Electrophoresis, Polyacrylamide GelABSTRACT
Abstract Endophytic fungi belonging to the genus Muscodor now transferred to Induratia are known producers of bioactive volatile organic compounds (VOCs) with many industrial applications. However, the members of this genus have rarely been reported to produce non-volatile metabolites including enzyme. Enzymes of the endophytes are degraders of the polysaccharides available in the host plants and the knowledge of enzyme production by Induratia spp. may provide insights into their possible biotechnological applications. The aim of this study was to evaluate the activity of amylase, cellulase, lipase, pectinase, phytase, protease, endo β-1,4 glucanase and exo β-1,4 glucanase enzymes produced by fungi of the species Induratia coffeana, Induratia yucatanensis and Induratia sp. isolated from organic coffee plants. All Induratia spp. were able to produce the extracellular enzymes cellulase, pectinase, protease, and phytase. Eight fungi were able to produce lipase and four produced amylase. The specific activity of endo β-1, 4 glucanase and exo β-1,4 glucanase enzymes were detected for 9 and 8 endophytic fungi, respectively. This work demonstrated for the first time, the array of enzymes produced by Induratia spp. isolated from Coffea arabica in organic systems in Brazil.
Subject(s)
Coffea/microbiology , Enzyme Activation , Volatile Organic Compounds/metabolism , Endophytes/enzymology , BrazilABSTRACT
Abstract We investigated the in vitro acaricide activity of the methanolic extract (ME) and alkaloid-rich fraction (AF) of Prosopis juliflora on Rhipicephalus microplus and correlated this effect with acetylcholinesterase (AChE) inhibition. The acaricide activity was evaluated using adult and larval immersion tests. Also, we studied the possible interaction mechanism of the major alkaloids present in this fraction via molecular docking at the active site of R. microplus AChE1 (RmAChE1). Higher reproductive inhibitory activity of the AF was recorded, with effective concentration (EC50) four times lower than that of the ME (31.6 versus 121 mg/mL). The AF caused mortality of tick larvae, with lethal concentration 50% (LC50) of 13.8 mg/mL. Both ME and AF were seen to have anticholinesterase activity on AChE of R. microplus larvae, while AF was more active with half-maximal inhibitory concentration (IC50) of 0.041 mg/mL. The LC-MS/MS analyses on the AF led to identification of three alkaloids: prosopine (1), juliprosinine (2) and juliprosopine (3). The molecular docking studies revealed that these alkaloids had interactions at the active site of the RmAChE1, mainly relating to hydrogen bonds and cation-pi interactions. We concluded that the alkaloids of P. juliflora showed acaricide activity on R. microplus and acted through an anticholinesterase mechanism.
Resumo A atividade carrapaticida in vitro do extrato metanólico (EM) e da fração de alcaloides (FA) de Prosopis juliflora foi investigada, frente ao Rhipicephalus microplus, e relacionada com a inibição da enzima acetilcolinesterase (AChE). A predição in silico das interações de alcaloides dessa fração com a AChE1 de R. microplus (RmAChE1) foi realizada por acoplamento molecular. A atividade carrapaticida foi avaliada, utilizando-se os ensaios de imersão de adultos e larvas. Maior efeito sobre parâmetros reprodutivos de teleóginas foi verificado para a FA, com valor de Concentração Efetiva 50% (CE50) (31.6 mg/mL), quatro vezes menor do que o valor do EM (121 mg/mL). A FA induziu mortalidade de larvas (Concentração Letal de 50% - CL50 = 13,8 mg/mL). A inibição da atividade da AChE de larvas do carrapato foi observada para EM e FA, sendo a FA mais ativa (Concentração Inibitória 50%- CI50 de 0,041mg/mL). As análises químicas da FA permitiram a identificação dos alcaloides prosopina (1), juliprosinina (2) e juliprosopina (3). No ensaio in silico, observou-se que esses alcaloides podem interagir com o sítio ativo da RmAChE1, principalmente por ligações de hidrogênio e interações cátion-pi. Os alcaloides de P. juliflora têm atividade carrapaticida contra R. microplus, atuando através do mecanismo anticolinesterásico.
Subject(s)
Animals , Plant Extracts/pharmacology , Cholinesterases/metabolism , Prosopis/chemistry , Rhipicephalus/drug effects , Rhipicephalus/enzymology , Alkaloids/pharmacology , Chromatography, Liquid , Enzyme Activation/drug effects , Tandem Mass Spectrometry , Acaricides/pharmacology , Molecular Docking Simulation , LarvaABSTRACT
Resumen: Los desórdenes congénitos de glicosilación son un conjunto de defectos genéticos de tipo multisistémico que afectan la función de la proteína. Se han descrito cerca de 75 enfermedades desde sus primeros estudios. En el presente estudio se desarrolló un método microespectrofotométrico para el diagnóstico de la enzima citosólica fosfomanosa isomerasa EC 5.3.1.8 (PMI), se analizaron 32 muestras de individuos con rango de edad de 0,6 a 27 años y se estableció el intervalo y el valor de referencia de actividad enzimática específica. Este estudio permitirá iniciar el diagnóstico de pacientes deficientes de la PMI de forma temprana y oportuna, lo cual la convierte en una posible enzima candidata para pruebas de tamizaje neonatal, ya que esta patología tiene un tratamiento fácil y de bajo costo, que consiste en la suplementación de manosa en forma oral. El diagnóstico clínico de este desorden metabólico beneficiará al paciente y a su familia al mejorar su calidad de vida, como también al sistema de salud colombiano.
Abstract: Congenital glycosylation disorders are a set of multi-systemic genetic defects affecting protein function. About 75 diseases have been described since early studies. This study developed a microspectrophotometric method for the diagnosis of the cytosolic enzyme phosphomannose isomerase (PMI) (EC 5.3.1.8), analyzed 32 samples of individuals ranging between 0.6 and 27 years old, and established the interval and reference value of specific enzyme activity. This study will allow early and timely diagnosis of PMI deficient patients, which makes this enzyme a potential candidate for neonatal screening tests since this pathology has an easy, low-cost treatment (oral administration of mannose supplements). Clinical diagnosis of this metabolic disorder will benefit the patient and his family by improving his quality of life, as well as the Colombian healthcare system.
Resumo: Os defeitos congénitos de glicosilação são um conjunto de defeitos genéticos de tipo mul-tissistêmico que afetam a função da proteína. Foram descritas 75 doenças desde seus primeiros estudos. Neste estudo, foi desenvolvido um método microespectrofotométrico para diagnosticar a enzima citosólica fosfomanose isomerase EC 5.3.1.8 (PMI); foram analisadas 32 amostras de indivíduos entre 0,6 e 27 anos e estabelecidos o intervalo e o valor de referência de atividade enzimática específica. Este estudo permitirá iniciar o diagnóstico de pacientes deficientes da PMI de forma precoce e oportuna, o que a converte em uma possível enzima candidata para testes genéticos de rastreio pré-natal, já que essa patologia tem um tratamento fácil e de baixo custo, que consiste na suplementação de manose por via oral. O diagnóstico clínico desse defeito metabólico beneficiará o paciente e sua família ao melhorar a qualidade de vida e o sistema de saúde colombiano.
Subject(s)
Humans , Infant , Child, Preschool , Child , Adolescent , Young Adult , Microspectrophotometry , Mannose-6-Phosphate Isomerase , Congenital Disorders of Glycosylation , Diagnosis , Enzyme ActivationABSTRACT
Adenosine 5'-monophosphate-activated protein activated protein kinase (AMPK), a heterotrimeric complex, is an important kinase to regulate glycolipid metabolism and energy balance involved in a variety physiological processes in human body. Many research indicated that the function and activity of AMPK were closely related to inflammation, diabetes and cancers. Recent reports show that inhibition of metformin (a first-line drug) on hepatic glucose in patients with hyperglycemia is associated with AMPK pathway, suggesting that targeting AMPK may be one of the effective strategies for the prevention and treatment of a variety of chronic diseases. Here, we review research progress on the structure, activation and regulation of AMPK in glycolipid metabolism to provide an insight into the basic and clinical research of diabetes therapy.
Subject(s)
AMP-Activated Protein Kinases , Adenosine , Adenosine Monophosphate , Energy Metabolism , Enzyme Activation , Glycolipids , HumansABSTRACT
The cyanobacterial circadian clock has three relatively independent parts: the input path, the core oscillator, and the output path. The core oscillator is composed of three clock proteins: KaiA, KaiB, and KaiC. The interactions among these three proteins generate a rhythmic signal and convey the input signals to the output signals to maintain the accuracy and stability of the oscillation of downstream signals. Based on the cyanobacterial circadian clock and the structure, function, and interaction of the clock proteins of the core oscillator, combining the recent results from our laboratory, this review summarized the recent progresses of the molecular mechanism of KaiA in regulating KaiC's enzymatic activity, mediating phase reset of the oscillator, and competing with CikA for the binding site of KaiB.
Subject(s)
Bacterial Proteins , Genetics , Metabolism , Circadian Clocks , Genetics , Circadian Rhythm Signaling Peptides and Proteins , Metabolism , Cyanobacteria , Genetics , Enzyme Activation , GeneticsABSTRACT
AMP-activated protein kinase (AMPK) is a key enzyme in the regulation of cellular energy homeostasis. Recent studies demonstrated that AMPK also plays an important role in the modulation of inflammation, an energy-intensive molecular response. The commonly used AMPK activators include 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) and A-769662. In addition, the biological activities of metformin and adiponectin are closely related to activation of AMPK. Numerous studies have shown that these AMPK activators play an effectively protective role in animal models of acute lung injury, asthma, colitis, hepatitis, atherosclerosis and other inflammatory diseases. Therefore, AMPK activators may have promising potential for the prevention and treatment of inflammation related diseases.
Subject(s)
AMP-Activated Protein Kinases , Physiology , Adiponectin , Pharmacology , Aminoimidazole Carboxamide , Pharmacology , Animals , Enzyme Activation , Inflammation , Metformin , Pharmacology , Pyrones , Pharmacology , Thiophenes , PharmacologyABSTRACT
Rifamycins, a group of bacterial RNA polymerase inhibitors, are the firstline antimicrobial drugs to treat tuberculosis. In light of the emergence of rifamycinresistant bacteria, development of new RNA polymerase inhibitors that kill rifamycinresistant bacteria with high bioavailability is urgent. Structural analysis of bacterial RNA polymerase in complex with inhibitors by crystallography and cryo-EM indicates that RNA polymerase inhibitors function through five distinct molecular mechanisms:inhibition of the extension of short RNA; competition with substrates; inhibition of the conformational change of the'bridge helix'; inhibition of clamp opening;inhibition of clamp closure. This article reviews the research progress of these five groups of RNA polymerase inhibitors to provide references for the modification of existing RNA polymerase inhibitors and the discovery of new RNA polymerase inhibitors.
Subject(s)
Antitubercular Agents , Therapeutic Uses , Bacteria , DNA-Directed RNA Polymerases , Metabolism , Drug Discovery , Drug Resistance, Bacterial , Enzyme Activation , Enzyme Inhibitors , Pharmacology , Humans , RNA, Bacterial , Tuberculosis , Drug TherapyABSTRACT
OBJECTIVE@#To investigate the effects of millimeter wave (MMW) exposure on apoptosis of human melanoma A375 cells and explore the mechanisms.@*METHODS@#Through electromagnetic field calculation we simulated MMW exposure in cells and calculated the specific absorption rate (SAR). The optimal irradiation parameters were determined according to the uniformity and intensity of the SAR. A375 cells were then exposed to MMV for 15, 30, 60, or 90 min, with or without pretreatment with the caspase-3 inhibitor AC-DEVD-fmk (10 μmol/L) for 1 h at 90 min before the exposure. CCK-8 assay was used to assess the changes in the viability and Annexin-V/ PI staining was used to detect the apoptosis of the cells following the exposures; Western blotting was used to detect the expression of caspase-3 in the cells.@*RESULTS@#The results of electromagnetic field calculation showed that for optimal MMV exposure, the incident field needed to be perpendicular to the bottom of the plastic Petri dish with the antenna placed below the dish. CCk-8 assay showed that MMW exposure significantly inhibited the cell viability in a time-dependent manner ( < 0.05); exposures for 15, 30, 60, and 90 min all resulted in significantly increased apoptosis of the cells ( < 0.05). The cells with MMW exposure showed significantly increased expression of caspase-3. The inhibitory effect of MMW on the cell viability was antagonized significantly by pretreatment of the cells with AC-DEVD-fmk ( < 0.05), which increased the cell viability rate from (36.7±0.09)% to (59.8±0.06)% ( < 0.05).@*CONCLUSIONS@#35.2 GHz millimeter wave irradiation induces apoptosis in A375 cells by activating the caspase-3 protein.
Subject(s)
Apoptosis , Caspase 3 , Metabolism , Caspase Inhibitors , Pharmacology , Cell Line, Tumor , Cell Survival , Electromagnetic Fields , Enzyme Activation , Humans , Magnetic Field Therapy , Melanoma , Pathology , Therapeutics , Time FactorsABSTRACT
Wolf cichlid, Parachromis dovii, is a species with a high potential for aquaculture in Central America; however, the knowledge of the digestive physiology in larvae period is limited. For these reason, this study evaluated the changes on digestive enzymes (alkaline and acid proteases, trypsin, chymotrypsin, aminopeptidase, carboxypeptidase, lipases, amylases, and phosphatases) during early ontogeny by biochemical analysis. All digestive enzymes were detected at first feeding (6 days after hatching, DAH, 9.49 mm, 168 degree-days DD). Afterwards all enzymes reached two main peaks in activity at 14 or 22 DAH (15.10 mm, 364 DD and 20.83 mm, 550 DD, respectively). Later, there was a gradual decrease in activity for trypsin and acid and alkaline phosphatases until reach the lowest values at 41 DAH. In the case of acid proteases, chymotrypsin, aminopeptidase, carboxypeptidase, lipase and amylase, all activities reached their maximum values at the end of the larval period, except for alkaline proteases, which showed the maximum value at 14 DAH (15.10 mm, 364 DD). Parachromis dovii larvae have an early capability to hydrolyze exogenous food, agreeing with other carnivorous neotropical cichlid species, for this reason we proposed that the weaning process could begin at 14 DAH.(AU)
El guapote lagunero (Parachromis dovii) es una especie con un alto potencial para la acuicultura en la región de América Central; sin embargo, existe un conocimiento limitado sobre la capacidad digestiva en el periodo larval. Por este motivo, este estudio evaluó los cambios de las enzimas digestivas (proteasas alcalinas y ácidas, tripsina, quimotripsina, aminopeptidasa, carboxipeptidasa, lipasas, amilasas y fosfatasas) durante la ontogenia temprana mediante análisis bioquímico. Todas las enzimas digestivas analizadas se detectaron en la primera alimentación (6 días después de la eclosión, DAH, 9.49 mm, 168 día-grados DD). Después, todas las enzimas alcanzaron dos picos máximos a los 14 o 22 DAH (15.10 mm, 364 DD and 20.83 mm, 550 DD, respectivamente). Después las actividades tripsina, fosfatasas ácidas y alcalina disminuyeron a sus valores más bajos a los 41 DAH. En el caso de las proteasas ácidas y alcalinas, quimotripsina, aminopeptidasa, carboxipeptidasa, lipasa y amilasa, los niveles de actividad aumentaron y alcanzaron su máximo valor al final del período larvario, excepto las proteasas alcalinas, que mostraron su máximo valor a los 14 DAH (15.10 mm, 364 DD). Las larvas de P. dovii tienen una capacidad temprana para hidrolizar alimentos exógenos, lo que concuerda con otras especies de cíclidos neotropicales carnívoros, por lo que proponemos que el proceso de destete inicie a los 14 DAH.(AU)
Subject(s)
Animals , Peptide Hydrolases/chemical synthesis , Cichlids/physiology , Enzyme Activation , AquacultureABSTRACT
Abstract Purpose: To investigate the effect of alprostadil on myocardial ischemia/reperfusion (I/R) in rats. Methods: Rats were subjected to myocardial ischemia for 30 min followed by 24h reperfusion. Alprostadil (4 or 8 μg/kg) was intravenously administered at the time of reperfusion and myocardial infarct size, levels of troponin T, and the activity of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) in the serum were measured. Antioxidative parameters, nitric oxide (NO) content and phosphorylated endothelial nitric oxide synthase 3 (p-eNOS) expression in the left ventricles were also measured. Histopathological examinations of the left ventricles were also performed. Results: Alprostadil treatment significantly reduced myocardial infarct size, serum troponin T levels, and CK-MB and LDH activity (P<0.05). Furthermore, treatment with alprostadil significantly decreased malondialdehyde (MDA) content (P<0.05) and markedly reduced myonecrosis, edema and infiltration of inflammatory cells. Superoxide dismutase and catalase activities (P<0.05), NO level (P<0.01) and p-eNOS (P<0.05) were significantly increased in rats treated with alprostadil compared with control rats. Conclusion: These results indicate that alprostadil protects against myocardial I/R injury and that these protective effects are achieved, at least in part, via the promotion of antioxidant activity and activation of eNOS.
Subject(s)
Animals , Male , Alprostadil/pharmacology , Myocardial Reperfusion Injury/prevention & control , Nitric Oxide Synthase Type III/metabolism , Antioxidants/pharmacology , Superoxide Dismutase/analysis , Myocardial Reperfusion Injury/metabolism , Myocardial Reperfusion Injury/pathology , Catalase/analysis , Random Allocation , Blotting, Western , Reproducibility of Results , Treatment Outcome , Rats, Sprague-Dawley , Oxidative Stress/drug effects , Troponin T/drug effects , Troponin T/blood , Enzyme Activation/drug effects , Creatine Kinase, MB Form/drug effects , Creatine Kinase, MB Form/blood , Heart Ventricles/drug effects , Heart Ventricles/pathology , L-Lactate Dehydrogenase/drug effects , L-Lactate Dehydrogenase/blood , Malondialdehyde/analysis , Myocardial Infarction/pathology , Nitric Oxide/analysisABSTRACT
Resumen La leucemia linfocítica aguda es la enfermedad oncológica con mayor incidencia en la población pediátrica, tanto a nivel mundial como en Costa Rica. Para su tratamiento requiere protocolos de quimioterapia complejos, lo que representa un reto constante para los médicos, ya que deben equilibrar los riesgos y beneficios del manejo. Es necesario tomar en cuenta los factores de riesgo de cada paciente, el grado de severidad de la enfermedad y los potenciales efectos adversos del tratamiento. A continuación, se reporta un caso de pancreatitis aguda edematosa no biliar, secundaria al uso de L-asparginasa, en un paciente con diagnóstico de leucemia linfocítica aguda, atendido en el Hospital Nacional de Niños "Dr. Carlos Sáenz Herrera". El paciente, quien se encontraba cumpliendo el régimen poliquimioterapeútico AHOPCA 2008, presentó clínica sugestiva de pancreatitis aguda en el día 50 de este, por lo que se decidió no colocar la quimioterapia indicada e inmediatamente se trasladó al Servicio de Emergencias. El cuadro clínico estaba asociado a laboratorios y ultrasonido anormales, por lo que fue tratado interdisciplinariamente y su pronóstico fue favorable; actualmente continúa con tratamiendo quimioterapeútico, como fue indicado.
Abstract The acute lymphocytic leukemia is the oncological disease with the highest incidence in the pediatric population both worldwide and in Costa Rica. It requires complex chemotherapy protocols, which confers a constant challenge on physicians to balance the risks and benefits of management. Therefore, it is necessary to take into account the risk factors of each patient, the degree of severity of the disease and the potential adverse effects of the treatment. A case report is presented with an acute non-biliary edematous pancreatitis, secondary to the use of L-asparaginase in a patient diagnosed with acute lymphocytic leukemia, seen at the National Children's Hospital "Dr. Carlos Sáenz Herrera". The patient who was started on the AHOPCA 2008 polychemotherapy regimen presented symptoms suggestive of acute pancreatitis on the day 50 of the same, so it was decided not to apply the indicated chemotherapy and transfer the patient to the Emergency Room. The clinical picture was associated with abnormal laboratories and ultrasound, so it was immediately treated interdisciplinarily, which is why its prognosis was favorable and currently he continues with chemotherapy treatment as indicated.
Subject(s)
Humans , Pancreatitis/chemically induced , Precursor Cell Lymphoblastic Leukemia-Lymphoma/complications , Enzyme ActivationABSTRACT
ABSTRACT The aim of this study was to optimize the dextranase production by fungus Pochonia chlamydosporia (VC4) and evaluate its activity in dextran reduction in sugarcane juice. The effects, over the P. chlamydosporia dextranase production, of different components from the culture medium were analyzed by Plackett-Burman design and central composite design. The response surface was utilized to determine the levels that, among the variables that influence dextranase production, provide higher production of these enzymes. The enzymatic effect on the removal of dextran present in sugarcane juice was also evaluated. It was observed that only NaNO3 and pH showed significant effect (p<0.05) over dextranase production and was determined that the levels which provided higher enzyme production were, respectively, 5 g/L and 5.5. The dextranases produced by fungus P. chlamydosporia reduced by 75% the dextran content of the sugarcane juice once treated for 12 hours, when compared to the control treatment.
Subject(s)
Models, Statistical , Saccharum/metabolism , Dextranase/biosynthesis , Hypocreales/enzymology , Temperature , Dextrans/metabolism , Culture Media/metabolism , Electrophoresis, Polyacrylamide Gel , Enzyme Activation , Fruit and Vegetable Juices/analysis , Chemical Fractionation/methods , Hydrogen-Ion Concentration , NitratesABSTRACT
Gnaphalium affine D. Don, a medicinal and edible plant, has been used to treat gout in traditional Chinese medicine and popularly consumed in China for a long time. A detailed phytochemical investigation on the aerial part of G. affine led to the isolation of two new esters of caffeoylquinic acid named (-) ethyl 1, 4-di-O-caffeoylquinate (1) and (-) methyl 1, 4-di-O-caffeoylquinate (2), together with 35 known compounds (3-37). Their structures were elucidated by spectroscopic data and first-order multiplet analysis. All the isolated compounds were tested for their xanthine oxidase inhibitory activity with an in vitro enzyme inhibitory screening assay. Among the tested compounds, 1 (IC 11.94 μmol·L) and 2 (IC 15.04 μmol·L) showed a good inhibitory activity. The current results supported the medical use of the plant.
Subject(s)
Adenine , Chemistry , Drugs, Chinese Herbal , Chemistry , Pharmacology , Enzyme Activation , Flavonoids , Chemistry , Gnaphalium , Chemistry , Gout Suppressants , Chemistry , Pharmacology , Hydroxybenzoates , Chemistry , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Phytochemicals , Chemistry , Pharmacology , Plant Components, Aerial , Chemistry , Plant Extracts , Chemistry , Pharmacology , Quinic Acid , Chemistry , Xanthine OxidaseABSTRACT
Hyperphosphorylated tau is the major protein component of neurofibrillary tangles in the brains of patients with Alzheimer's disease (AD). However, the mechanism underlying tau hyperphosphorylation is not fully understood. Here, we demonstrated that exogenously expressed wild-type human tau40 was detectable in the phosphorylated form at multiple AD-associated sites in cytoplasmic and nuclear fractions from HEK293 cells. Among these sites, tau phosphorylated at Thr205 and Ser214 was almost exclusively found in the nuclear fraction at the conditions used in the present study. With the intracellular tau accumulation, the Ca concentration was significantly increased in both cytoplasmic and nuclear fractions. Further studies using site-specific mutagenesis and pharmacological treatment demonstrated that phosphorylation of tau at Thr205 increased nuclear Ca concentration with a simultaneous increase in the phosphorylation of Ca/calmodulin-dependent protein kinase IV (CaMKIV) at Ser196. On the other hand, phosphorylation of tau at Ser214 did not significantly change the nuclear Ca/CaMKIV signaling. Finally, expressing calmodulin-binding protein-4 that disrupts formation of the Ca/calmodulin complex abolished the okadaic acid-induced tau hyperphosphorylation in the nuclear fraction. We conclude that the intracellular accumulation of phosphorylated tau, as detected in the brains of AD patients, can trigger nuclear Ca/CaMKIV signaling, which in turn aggravates tau hyperphosphorylation. Our findings provide new insights for tauopathies: hyperphosphorylation of intracellular tau and an increased Ca concentration may induce a self-perpetuating harmful loop to promote neurodegeneration.
Subject(s)
Alzheimer Disease , Metabolism , Pathology , Calcium , Metabolism , Calcium-Calmodulin-Dependent Protein Kinase Type 4 , Metabolism , Cell Nucleus , Metabolism , Enzyme Activation , Physiology , HEK293 Cells , Humans , Neurons , Metabolism , Pathology , Phosphorylation , Signal Transduction , Physiology , tau Proteins , MetabolismABSTRACT
Gnaphalium affine D. Don, a medicinal and edible plant, has been used to treat gout in traditional Chinese medicine and popularly consumed in China for a long time. A detailed phytochemical investigation on the aerial part of G. affine led to the isolation of two new esters of caffeoylquinic acid named (-) ethyl 1, 4-di-O-caffeoylquinate (1) and (-) methyl 1, 4-di-O-caffeoylquinate (2), together with 35 known compounds (3-37). Their structures were elucidated by spectroscopic data and first-order multiplet analysis. All the isolated compounds were tested for their xanthine oxidase inhibitory activity with an in vitro enzyme inhibitory screening assay. Among the tested compounds, 1 (IC 11.94 μmol·L) and 2 (IC 15.04 μmol·L) showed a good inhibitory activity. The current results supported the medical use of the plant.
Subject(s)
Adenine , Chemistry , Drugs, Chinese Herbal , Chemistry , Pharmacology , Enzyme Activation , Flavonoids , Chemistry , Gnaphalium , Chemistry , Gout Suppressants , Chemistry , Pharmacology , Hydroxybenzoates , Chemistry , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Phytochemicals , Chemistry , Pharmacology , Plant Components, Aerial , Chemistry , Plant Extracts , Chemistry , Pharmacology , Quinic Acid , Chemistry , Xanthine OxidaseABSTRACT
Abstract Purpose: To investigate whether modulating GSK-3β could attenuate myocardial ischemia reperfusion injury (MIRI) induced acute lung injury (ALI) and analyze the underlying mechanism. Methods: Male SD rats were subjected to MIRI with or without myocardial ischemic post-conditioning in the presence or absence of GSK-3β inhibitor. GSK-3β inhibitor was injected peritoneally 10min before MIRI. Lung W/D weight ratio, MPO, PMNs, histopathological changes, TUNEL, Bax, Bcl-2, IL-6, IL-8, IL-10, GSK-3β, and caspase-3 were evaluated in the lung tissues of all rats. Results: After MIRI, lung injury was significantly increased manifested as significant morphological changes and increased leukocytes in the interstitial capillaries, Lung W/D ratio, MPO, and PMN in BALF, which was associated with enhanced inflammation evidenced by increased expressions of IL-6, IL-8 and reduced expression of IL-10. MIRI significantly increased cell apoptosis in the lung as increased levels of apoptotosis, Bax, cleaved caspase-3, and reduced expression of Bcl-2 was observed, which was concomitant with reduced p-GSK-3β. All these changes were reversed/prevented by ischemic post-conditioning, while these beneficial effects of ischemic post-conditioning were abolished by GSK-3β inhibition. Conclusion: Myocardial ischemia reperfusion injury induces acute lung injury by induction of inflammation and cell apoptosis. Ischemic post-conditioning protects the lung from ALI following MIRI by increasing p-GSK-3β.
Subject(s)
Animals , Male , Myocardial Reperfusion Injury/prevention & control , Protective Agents/metabolism , Acute Lung Injury/prevention & control , Ischemic Postconditioning/methods , Glycogen Synthase Kinase 3 beta/metabolism , Random Allocation , Down-Regulation , Interleukins/metabolism , Rats, Sprague-Dawley , Apoptosis/drug effects , Peroxidase/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Protective Agents/pharmacology , In Situ Nick-End Labeling , Models, Animal , Enzyme Activation , bcl-2-Associated X Protein/metabolism , Caspase 3/metabolism , Acute Lung Injury/enzymology , Glycogen Synthase Kinase 3 beta/antagonists & inhibitors , Glycogen Synthase Kinase 3 beta/pharmacology , Inflammation/metabolism , Myocardial Infarction/pathology , Neutrophils/enzymologyABSTRACT
A fim de se agregar valor ao resíduo farelo de trigo gerado em indústrias do setor alimentício avaliou-se, no presente trabalho, o potencial deste subproduto como substrato para produção de enzima xilanase no cultivo em estado sólido, utilizando consórcios fúngicos bem como os fungos Aspergillus oryzae CCT nº 0975 (ATCC9362) eTrichoderma reesei CCT nº 2768 - QM 9414. Para tanto utilizou-se o farelo de trigo, não lavado e não autoclavado, como fonte de carbono e energia na fermentação em estado sólido pelo fungo Aspergillus oryzae que apresentou maior produção do percentual de proteína nas 72 horas de cultivo. Depois de realizado um Delineamento Composto Central Rotacional (DCCR) - planejamento fatorial 23,com três repetições no ponto central e seis pontos axiais - partiu-se para otimização dos fatores que foram considerados significativos no processo: umidade, pH e granulometria. Os fatores foram considerados significativos pela A NOVA com o nível de 95% de confiança e com o resultado otimizado de atividade enzimática de (1.84 ± 0.01) UI/mL utilizando pH 3,3, granulometria de 900,0 µm e umidade de 40%. O caldo enzimático obtido foi considerado eficiente na modificação de tipificação de farinhas de trigo pelo estudo dos parâmetros reológicos do falling number e alveografia sendo estável por cerca de 3 meses
This study aimed to find alternatives for wheat bran disposal destination generated in food industry sector,thus contributing to the reduction of the resultant impact of residue depo-sition in the environment. The poten-tial of the wheat bran as a substrate for xylanase production by solid-state fermentation using fungal con-sortiums as well as Aspergillus ory-zae (ATCC9362) and Trichoderma reesei (2768) was valued. The use of non-washed and non-autoclaved wheat bran as carbon and energy source in solid-state fermentation by A. oryzae fungus showed greater per-centage of produced protein after 72 h of cultivation. The use of a central composite rotatable design(CCRD), 23 factorial planning with three rep-etitions at the central point as well as six axial points, coupled with Sur-face Response Methodology (SRM) allowed to assay the influence of hu-midity, pH, and grain size (indepen-dent variables or factors) on the xy-lanase activity(dependent variable or response) as well as to optimize the best conditions for the enzyme production. The results showed that all factors and their combinations were significant at 95% confidence level. The optimized xylanase activi-ty was (1.84 ± 0.01) UI/mL, obtained at 40% humidity and pH 3.3 with a grain size of 900.0 µm. The produced broth was stable for 3 months and approximately had 50% of the initial xylanase activity at 4°C. SDS-PAGE assay showed that xylanase has 30 kDa molar mass. The obtained en-zymatic broth was efficient to modify wheat flours as shown by the falling number rheologic parameters and alveography assay