RESUMO
Hyperpolarization-activated cationic and cyclic nucleotide-gated channels (HCN) comprise four homologous subunits (HCN1-HCN4). HCN channels are found in excitable and non-excitable tissues in mammals. We have previously shown that HCN2 may transport ammonium (NH4 (+)), besides sodium (Na(+)), in the rat distal nephron. In the present work, we identified HCN1 and HCN3 in the proximal tubule (PT) and HCN3 in the thick ascending limb of Henle (TALH) of the rat kidney. Immunoblot assays detected HCN1 (130 kDa) and HCN3 (90 KDa) and their truncated proteins C-terminal HCN1 (93 KDa) and N-terminal HCN3 (65 KDa) in enriched plasma membranes from cortex (CX) and outer medulla (OM), as well as in brush-border membrane vesicles. Immunofluorescence assays confirmed apical localization of HCN1 and HCN3 in the PT. HCN3 was also found at the basolateral membrane of TALH. We evaluated chronic changes in mineral dietary on HCN3 protein abundance. Animals were fed with three different diets: sodium-deficient (SD) diet, potassium-deficient (KD) diet, and high-potassium (HK) diet. Up-regulation of HCN3 was observed in OM by KD and in CX and OM by HK; the opposite effect occurred with the N-terminal truncated HCN3 in CX (KD) and OM (HK). SD diet did not produce any change. Since HCN channels activate with membrane hyperpolarization, our results suggest that HCN channels may play a role in the Na(+)-K(+)-ATPase activity, contributing to Na(+), K(+), and acid-base homeostasis in the rat kidney.
Assuntos
Canais Disparados por Nucleotídeos Cíclicos Ativados por Hiperpolarização/metabolismo , Túbulos Renais Proximais/metabolismo , Alça do Néfron/metabolismo , Canais de Potássio/metabolismo , Potássio na Dieta/metabolismo , Animais , Membrana Celular/metabolismo , Hipopotassemia/patologia , Córtex Renal/metabolismo , Medula Renal/metabolismo , Masculino , Microvilosidades/metabolismo , Técnicas de Patch-Clamp , Ratos , Ratos Wistar , Sódio/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismoRESUMO
Due to the great global concern regarding bacterial resistance to antibiotics, an ongoing search for new molecules having antibacterial activity is necessary. This study evaluated the antibacterial and anticandidal effects of a hexane extract from the root of Salvia apiana. Salvia extracts at concentrations of 27, 13.5, 6.8 and 3.4mg/ml caused growth inhibition of Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis and Candida albicans. However, no significant effect was observed on Escherichia coli and Candida tropicalis in comparison to vehicle. It was here demonstrated for the first time that Salvia apiana has an important antimicrobial effect on human pathogens of great clinical value, thus opening the field to continue the evaluation of this lamiaceous plant for its future use as a therapeutic agent.
Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Candida/efeitos dos fármacos , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Salvia/química , Antibacterianos/isolamento & purificação , Antifúngicos/isolamento & purificação , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Hexanos , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Raízes de Plantas/química , Solventes , Especificidade da EspécieRESUMO
BACKGROUND: Cyclooxygenase-2 selective inhibitors have been developed to alleviate pain and inflammation; however, the use of a selective cyclooxygenase-2 inhibitor is associated with mild edema, hypertension, and cardiovascular risk. AIM: To evaluate, in an experimental model in normotensive rats, the effect of treatment with parecoxib in comparison with diclofenac and aspirin and L-NAME, a non-selective nitric oxide synthetase, on mean arterial blood pressure, and cyclooxygenase-1 and -2 messenger RNA and protein expression in aortic tissue. METHODS: Rats were treated for seven days with parecoxib (10 mg/kg/day), diclofenac (3.2 mg/kg/day), aspirin (10 mg/kg/day), or L-NAME (10 mg/kg/day). Mean arterial blood pressure was evaluated in rat tail; cyclooxygenase-1 and -2 were evaluated by reverse transcription-polymerase chain reaction and Western blot analysis in aortic tissue. RESULTS: Parecoxib and L-NAME, but not aspirin and diclofenac, increased mean arterial blood pressure by about 50% (p < 0.05) without changes in cardiac frequency. Messenger RNA cyclooxygenase-1 expression in aortic tissue was not modified with any drug (p < 0.05). L-NAME and parecoxib treatment decreased messenger RNA cyclooxygenase-2 and cyclooxygenase-2 (p < 0.05). While cyclooxygenase-1 protein decreased with the three drugs tested but not with L-NAME (p < 0.05), the cyclooxygenase-2 protein decreased only with aspirin and parecoxib (p < 0.05). CONCLUSION: Parecoxib increases the blood pressure of normotensive rats by the suppression of COX-2 gene expression, which apparently induced cardiovascular control.
Assuntos
Pressão Sanguínea/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase 2/toxicidade , Ciclo-Oxigenase 2/efeitos dos fármacos , Isoxazóis/toxicidade , Animais , Aorta/efeitos dos fármacos , Aorta/metabolismo , Aspirina/toxicidade , Western Blotting , Ciclo-Oxigenase 1/genética , Ciclo-Oxigenase 2/genética , Diclofenaco/toxicidade , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Masculino , NG-Nitroarginina Metil Éster/toxicidade , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos WKY , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The ATP-related compounds in whole blood or red blood cells have been used to evaluate the energy status of erythrocytes and the degradation level of the phosphorylated compounds under various conditions, such as chronic renal failure, drug monitoring, cancer, exposure to environmental toxics, and organ preservation. The complete interpretation of the energetic homeostasis of erythrocytes is only performed using the compounds involved in the degradation pathway for adenine nucleotides alongside the uric acid value. For the first time, we report a liquid chromatographic method using a diode array detector that measures all of these compounds in a small human whole blood sample (125 µL) within an acceptable time of 20 min. The stability was evaluated for all of the compounds and ranged from 96.3 to 105.1% versus the day zero values. The measurement had an adequate sensitivity for the ATP-related compounds (detection limits from 0.001 to 0.097 µmol/L and quantification limits from 0.004 to 0.294 µmol/L). This method is particularly useful for measuring inosine monophosphate, inosine, hypoxanthine, and uric acid. Moreover, this assay had acceptable linearity (r > 0.990), precision (coefficients of variation ranged from 0.1 to 2.0%), specificity (similar retention times and spectra in all samples) and recoveries (ranged from 89.2 to 104.9%). The newly developed method is invaluable for assessing the energetic homeostasis of red blood cells under diverse conditions, such as in vitro experiments and clinical settings.
Assuntos
Trifosfato de Adenosina/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Eritrócitos/química , Eritrócitos/fisiologia , Homeostase/fisiologia , Adenosina/análise , Adenosina/metabolismo , Contagem de Eritrócitos , Humanos , Concentração de Íons de Hidrogênio , Hipoxantina/análise , Hipoxantina/metabolismo , Inosina/análise , Inosina/metabolismo , Inosina Monofosfato/análise , Inosina Monofosfato/metabolismo , Sensibilidade e Especificidade , Ácido Úrico/análise , Ácido Úrico/metabolismoRESUMO
Background: Alzheimer's disease (AD) and type 2 diabetes mellitus (DM2) are chronic degenerative diseases with complex molecular processes that are potentially interconnected. The aim of this work was to predict the potential molecular links between AD and DM2 from different sources of biological information. Materials and Methods: In this work, data mining of nine databases (DisGeNET, Ensembl, OMIM, Protein Data Bank, The Human Protein Atlas, UniProt, Gene Expression Omnibus, Human Cell Atlas, and PubMed) was performed to identify gene and protein information that was shared in AD and DM2. Next, the information was mapped to human protein-protein interaction (PPI) networks based on experimental data using the STRING web platform. Then, gene ontology biological process (GOBP) and pathway analyses with EnrichR showed its specific and shared biological process and pathway deregulations. Finally, potential biomarkers and drug targets were predicted with the Metascape platform. Results: A total of 1,551 genes shared in AD and DM2 were identified. The highest average degree of nodes within the PPI was for DM2 (average = 2.97), followed by AD (average degree = 2.35). GOBP for AD was related to specific transcriptional and translation genetic terms occurring in neurons cells. The GOBP and pathway information for the association AD-DM2 were linked mainly to bioenergetics and cytokine signaling. Within the AD-DM2 association, 10 hub proteins were identified, seven of which were predicted to be present in plasma and exhibit pharmacological interaction with monoclonal antibodies in use, anticancer drugs, and flavonoid derivatives. Conclusion: Our data mining and analysis strategy showed that there are a plenty of biological information based on experiments that links AD and DM2, which could provide a rational guide to design further diagnosis and treatment for AD and DM2.
Assuntos
Doença de Alzheimer , Diabetes Mellitus Tipo 2 , Humanos , Doença de Alzheimer/genética , Diabetes Mellitus Tipo 2/genética , Mapas de Interação de Proteínas/genética , Biologia Computacional , Bases de Dados FactuaisRESUMO
Dactylopius opuntiae is an insect pest that contains at least carminic acid, which has antioxidant properties. Since there is a relationship between the antioxidant ability and preservative action of compounds applied to meat products, the purpose of this study was to evaluate the antioxidant activity and usefulness of a D. opuntiae extract for beef patty preservation. The insects were bred and processed to obtain a liquid extract. For the extract, its carminic acid content, antioxidant activity against two free radicals, and actions on food quality parameters were determined. The D. opuntiae dry powder contained 2.91% w/w carminic acid, while the liquid extract exhibited an IC50 value of 3437.8 ± 67.8 and 19633.0 ± 674.5 µg/mL against the DPPH and ABTS radicals. Nevertheless, these antioxidant actions were lower than those found in a D. coccus extract. The D. opuntiae extract improved in a short time the redness and yellowness, eliminated the unfavorable effect of their vehicle on the MetMb level, and greatly reduced the TBARS formation. For the first time, an extract of D. opuntiae was applied to beef patties, and its beneficial antioxidant action on meat acceptance parameters was confirmed, which has potential commercial applications.
RESUMO
The adjunctive use of fluoroquinolone (FQ) agents in patients with periodontitis produces contradictory results. There has been no meta-analysis performed based on the evaluations of FQ use that would enable making appropriate clinical decisions. Our study aimed to evaluate, via a systematic review and metaanalysis conducted in accordance with the PRISMA (Preferred Reporting Items for Systematic reviews and Meta-Analyses) guidelines, the clinical benefits, antimicrobial effects and safety profiles of the FQ agents administered to periodontitis patients under a conventional treatment regime. Relevant databases were searched for studies published up to May 2020, with the quality and risk of bias evaluations performed on the selected studies, and meta-analyses, funnel plots and heterogeneity tests carried out based on the obtained data. Any finding of p-value less than 0.05 was considered statistically significant. Quality and the risk of bias ranged from high to low. With acceptable heterogeneity and no reporting bias, the meta-analyses showed that local or systemic FQ use produced the following results: a reduced probing depth change (ΔPD) (p < 0.00001 at ≤3 months); reduced bleeding on probing (BOP) (p < 0.00001 at 3-6 months); reduced subgingival detection of Aggregatibacter actinomycetemcomitans for up to 12 months (p-values from <0.00001 to 0.001); and an insignificant number of adverse events (p ≥ 0.05) in patients subjected to a conventional therapy as compared to those subjected to an antibiotic-free therapy. Our study found evidence to show that FQ administration provides clinical benefits and ensures antibacterial effects in periodontitis patients subjected to a conventional therapy regime.
Assuntos
Fluoroquinolonas , Periodontite , Antibacterianos/uso terapêutico , Fluoroquinolonas/uso terapêutico , Humanos , Periodontite/tratamento farmacológicoRESUMO
AIMS: The purpose of this meta-analysis was to evaluate the clinical efficacy of non-steroidal anti-inflammatory drugs and dexamethasone on the trismus, postsurgical pain, facial swelling, as well as the analgesic consumption after third molar surgery. MATERIAL AND METHODS: The reports were identified in the most important medical databases. Those studies that met the requirements were fully assessed according to the inclusion and exclusion criteria. The quality of each report was evaluated with the Oxford Quality Scale and using the Cochrane Collaboration's risk of bias tool. Each meta-analysis was done using the technique of mean difference and 95% confidence intervals employing a random effects model with the Review Manager 5.3., from the Cochrane Library. Significant statistical difference was accepted when the p value was less than 0.05 on the test of overall effect (Z value). RESULTS: Qualitative evaluation was done using the data of 330 patients extracted from seven articles and the quantitative assessment with data of 200 patients from three reports. It was not observed difference among non-steroidal anti-inflammatory drugs and dexamethasone in any of the clinical effectiveness indicators. CONCLUSION: The outcomes of our meta-analysis indicate that non-steroidal anti-inflammatory drugs and dexamethasone have good therapeutic effect for the management of inflammatory complications following to third molar surgery.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Dexametasona/uso terapêutico , Dente Serotino/cirurgia , Extração Dentária/efeitos adversos , Analgésicos/uso terapêutico , Estudos de Avaliação como Assunto , Humanos , Dor Pós-Operatória/prevenção & controle , Trismo/prevenção & controleRESUMO
Objetive: To evaluate the correlation between salivary biomarkers (the salivary antioxidant ability, salivary level of polyphenols, and other antioxidants) with plaque-induced gingivitis exacerbated by pregnancy in pregnant and nonpregnant women. Material and Methods: For this observational study, medical records, dental examinations, and analyses of saliva samples were carried out in pregnant and nonpregnant women. A p-value of <0.05 was considered significant. Results: The pregnant women (n =17) exhibited a lower antioxidant capacity (p-value=0.0041), higher levels of polyphenols, gingival index, bleeding on probing, and subjects consuming mineral-enriched products (p-value from <0.0001 to 0.0466), and unchanged levels of phosphotungstic acid reactive substances, proteins, oral hygienic habits, plaque index and probing depth (p-value from 0.0683 to 0.8358), in comparison with the nonpregnant women (n=9). Also, a positive correlation between the gingival index and salivary polyphenol content was observed (r-value = 0.4087, p-value = 0.0202). Conclusion: The salivary polyphenols correlate with plaque-induced gingivitis exacerbated by pregnancy, suggesting a deficiency of salivary antioxidant protection.
Objetivo: Evaluar la correlación entre los biomarcadores salivales (la capacidad antioxidante salival, el nivel salival de polifenoles y otros antioxidantes) con la gingivitis inducida por placa exacerbada por el embarazo en mujeres embarazadas y no embarazadas. Material y Métodos: Para este estudio observacional, se realizaron registros médicos, exámenes dentales y análisis de muestras de saliva en mujeres embarazadas y no embarazadas. Se consideró significativo un valor de p<0,05. Resultados: Las gestantes (n=17) presentaron menor capacidad antioxidante (p=0,0041), mayores niveles de polifenoles, índice gingival, sangrado al sondaje y los sujetos que consumían productos enriquecidos con minerales (p<0,0001 a p<0,0466), y no hubo diferencias en los niveles de sustancias reactivas al ácido fosfotúngstico, proteínas, hábitos de higiene bucal, índice de placa y profundidad de sondaje (p=0,0683 a 0,8358), en comparación con las mujeres no embarazadas (n=19). Además, se observó una correlación positiva entre el índice gingival y elcontenido de polifenoles salivales (r = 0,4087, p= 0,0202). Conclusión: Los polifenoles salivales se correlacionan con la gingivitis inducida por placa y exacerbada por el embarazo, lo que sugiere una deficiencia de protección antioxidante salival.
Assuntos
Humanos , Feminino , Gravidez , Saliva/imunologia , Biomarcadores/análise , Gengivite/imunologia , Polifenóis , AntioxidantesRESUMO
OBJECTIVES: Levofloxacin monitoring is recommended to obtain clinical cure and low incidence of antimicrobial resistance. During the monitoring procedure, levofloxacin should be measured in plasma samples and several assays are reported for this purpose. However, those methods do not have all of the characteristics for an accessible and reliable drug monitoring. For this reason, we develop a method that has all of the essential characteristics for levofloxacin monitoring. DESIGN AND METHODS: The procedure of validation was done in terms of Food and Drug Administration guidelines. Subsequently, our assay was applied in plasma samples obtained from healthy volunteers with a single oral administration of levofloxacin as well as patients with respiratory diseases under levofloxacin therapy. RESULTS: Levofloxacin was extracted from samples using only two precipitation steps. Our assay had a rapid run time (5min), adequate sensitivity (0.05µg/ml of lower limit of quantification), and acceptable parameters of validation. Moreover, compound identities were supported using three dimensional spectra and purities were confirmed employing similarity factors (values>900). Variable concentrations of levofloxacin in samples were observed during the application. CONCLUSIONS: Levofloxacin is successfully quantified using our method that shows reliable results, appropriate range, rapid analyses, and cost-effective measurements under a simple and easy technique while all prior methods did not have it all together. Consequently, our method is a valuable tool for routine drug monitoring. Moreover, a complete evaluation of specificity was done for levofloxacin in plasma samples for the first time. Meanwhile, the application data supported the necessity of levofloxacin monitoring.
Assuntos
Antibacterianos/sangue , Monitoramento de Medicamentos/normas , Levofloxacino/sangue , Humanos , Reprodutibilidade dos TestesRESUMO
Fluoroquinolone agents are used widely for the treatment of infectious diseases which are a common cause of deaths around the world. The level of oxidative stress in patients taking fluoroquinolone antibiotics has not been considered a factor to reduce the clinical efficacy of this kind of drugs. Patients with diabetes and/or cardiovascular diseases present abnormal levels of oxidative stress in the blood stream. In this regards, our hypothesis supposes that patients with diabetes and/or cardiovascular disease suffering a bacterial disease could experience a therapeutic failure and bacterial resistance when treated with fluoroquinolones. The crucial mechanism could be an inefficient blood distribution of the drug via red blood cell dysfunction induced by oxidative stress that might reduce the pharmacokinetic-pharmacodinamic ratios. In this way, we review the scientific information to support our hypothesis alongside possible implications. Additionally, this work exhibits the urgent need of studies considering these conditions for quinolone agents.
Assuntos
Infecções Bacterianas/tratamento farmacológico , Doenças Cardiovasculares/tratamento farmacológico , Complicações do Diabetes/tratamento farmacológico , Diabetes Mellitus/tratamento farmacológico , Farmacorresistência Bacteriana , Fluoroquinolonas/uso terapêutico , Estresse Oxidativo , Trifosfato de Adenosina/metabolismo , Antibacterianos/uso terapêutico , Antioxidantes/metabolismo , Área Sob a Curva , Infecções Bacterianas/complicações , Doenças Cardiovasculares/complicações , Humanos , Modelos Teóricos , Transdução de SinaisRESUMO
Debido a la gran problemática mundial de la resistencia bacteriana a los antibióticos, es necesaria la búsqueda continua de nuevas moléculas con características antimicrobianas. Este estudio evaluó el efecto antibacteriano y antifúngico de un extracto hexánico proveniente de la raíz de Salvia apiana. Los extractos de salvia a las concentraciones de 27; 13,5; 6,8 y 3,4mg/ml causaron inhibición del crecimiento de Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis y Candida albicans. Sin embargo, no presentaron efecto significativo sobre Escherichia coli y Candida tropicalis al compararse con los valores del vehículo en las valoraciones de difusión en pozo. Se demostró que S. apiana tiene un efecto antimicrobiano significativo sobre patógenos de gran importancia clínica, lo que abre el campo para continuar evaluando a esta lamiácea en vistas a su posible empleo en el futuro como un agente terapéutico
Due to the great global concern regarding bacterial resistance to antibiotics, an ongoing search for new molecules having antibacterial activity is necessary. This study evaluated the antibacterial and anticandidal effects of a hexane extract from the root of Salvia apiana. Salvia extracts at concentrations of 27, 13.5, 6.8 and 3.4mg/ml caused growth inhibition of Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis and Candida albicans. However, no significant effect was observed on Escherichia coli and Candida tropicalis in comparison to vehicle. It was here demonstrated for the first time that Salvia apiana has an important antimicrobial effect on human pathogens of great clinical value, thus opening the field to continue the evaluation of this lamiaceous plant for its future use as a therapeutic agent