Ciências Farmacêuticas
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- ItemAvaliação da eficácia de atropina e pralidoxima na intoxicação aguda por clorpirifós e desenvolvimento de metodologia analítica para identificação e quantificação de organofosforados e seus antídotos por HPLC-MS/MS(Universidade Federal do Espírito Santo, 2016-03-22) Marques, Graziany Leite Moreira; Pelição, Fabrício Souza; Sampaio, Karla Nívea; Pires, Rita Gomes Wanderley; Martinis, Bruno Spinosa deOrganophosphorus insecticides (OP) are probably the most widely used pesticides in the world. In Brazil, these compounds are considered one of the main responsible for poisoning. Chlorpyrifos (CPF), an OP compound highly used, promotes, similarly to other OP, inhibition of the cholinesterases (ChEs) leading to the accumulation of acetylcholine in the central and peripheral cholinergic synapses. The treatment for OP poisoning involves a combination of muscarinic antagonists, such as atropine (ATR); oximes for reactivating the ChEs, mainly represented by the pralidoxime (2-PAM) and benzodiazepines, generally diazepam, for treatment of occasional seizures. However, the effectiveness of oximes in humans is still doubtful, mainly due to the polarity of these compounds, which leads to poor penetration of these drugs into the central nervous system (CNS). Therefore, this study aims to investigate the efficacy of the antidotes adopted in the OP poisoning, over the clinical signs and on the reactivation of the ChEs inhibition induced by the acute poisoning with CPF, as well as validate analytical method for identifying and quantifying various OPs, ATR and 2-PAM, using high-performance liquid chromatography coupled to mass spectrometry (HPLCMS/MS). Adult male rats (n=280) were divided into 8 groups: saline (SAL), CPF, SAL+ATR, CPF+ATR, SAL+2-PAM, CPF+2-PAM, SAL+ATR+2-PAM and CPF+ATR+2-PAM. The antidotes treatment with ATR (10 mg/kg); 2-PAM (40 mg/kg) or ATR (10mg/kg) +2-PAM (40 mg/kg) was delivered by intraperitoneal injections (i.p.) one hour after the administration of CPF (30 mg/kg, i.p.) or SAL (0.9%, i.p.) and the animals were observed for acute toxicity signs for up to 5 hours. In periods of 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours and 24 hours after the antidotes treatment administration, the animals were decapitated for collecting blood and brain (except cerebellum) samples for measurements of ChEs activity, as well as for CPF and CPF-oxon analysis by HPLC-MS/MS. The method developed for the OPs and antidotes analysis proved to be efficient and fast, attending the validation criteria stated in the RDC nº 27 of ANVISA (BRASIL, 2012). The analysis demonstrates that the CPF blood concentration significantly decreased over the first 24 hours after administration. The CPF caused inhibition of plasma ChE and treatment with 2-PAM was capable of reversing this inhibition for up to 1 hour after administration. However, when considering the brain AChE the 2-PAM only induced a small reactivation of the enzyme from the second hour of the treatment administration. Although not fully effective, the treatments with ATR, 2-PAM or ATR+2-PAM employed prevented or reverted, at different levels, the typical cholinergic symptoms of the OP poisoning.
- ItemAvaliação da influência do acompanhamento farmacoterapêutico e da terapia insulínica sobre a qualidade de vida, adesão ao tratamento e estresse oxidativo em pacientes com diabetes mellitus tipo 2(Universidade Federal do Espírito Santo, 2018-06-29) Meriguete, Angélica Marchesi Lira; Guimarães do Bem, Daniela Amorim Melgaço; Gonçalves, Rita de Cássia Ribeiro; Zandonade, ElianaThe practices of Pharmaceutical Care in Brazil are structured to promote medication adherence and quality of life, especially relevant in the context of type 2 diabetes mellitus (T2DM), which represents a growing problem of global health. It is a chronic metabolic disorder characterized by an increase in oxidative stress, which is related to the appearance of its main complications. T2DM is related to long and complex treatments, presence of comorbidities and complications which, together interfere negatively in the adherence to therapy. There are reports in the literature that the application of pharmaceutical care services has brought benefits on metabolic control and improved medication adherence of individuals with type 2 diabetes. In this study, it was evaluated if the pharmacotherapeutic follow-up performed for six months in patients with type 2 diabetes mellitus interferes in metabolic control, treatment adherence, quality of life and levels of pro and antioxidant species, besides the influence of insulin therapy on these parameters. Data collection was conducted before and after the six months of follow-up. 75 patients completed the study, 44 in the non-insulin therapy group and 31 in the insulin therapy group. After follow-up, an improvement in medication adherence was observed in the two groups. Similarly, there was a significant increase in the quality of life scores (p<0.0001) for the total sample. The physical domain of the insulin therapy group showed no change between the period (p = 0.052). In the oxidative stress parameters evaluated, there was a decrease in nitric oxide levels in both groups (p<0.0001) and an increase in superoxide dismutase concentration for the group with insulin therapy (p=0.0048). These results show that the proposed pharmacotherapeutic followup positively influenced on the medication adherence, quality of life and oxidative stress levels, and therefore, may contribute to delay the onset of the main chronic complications of the disease
- ItemAvaliação do efeito gastroprotetor das sementes de Persea americana Mill(Universidade Federal do Espírito Santo, 2018-03-28) Athaydes, Brena Ramos; Kitagawa, Rodrigo Rezende; Gonçalves, Rita de Cássia Ribeiro; Borges, Warley de Souza; Fronza, MarcioPeptic ulcer is one of the most common diseases affecting the world's population. It is characterized by an imbalance between protective (mucus and bicarbonate production, antioxidants and prostaglandins) and aggressor factors (oxidative stress, non-steroidal anti-inflammatory drugs and Helicobacter pylori) of the gastric mucosa. The inflammatory ulcer process induces the release of pro-inflammatory cytokines and oxidative stress, which can chronically lead to gastric cancer. In Brazil, there are reports of the use of Persea americana Mill. seeds (avocado) for the treatment of gastric diseases, however, without scientific evidence. Some research also highlights its antioxidant and antimicrobial effects. In this context, we evaluated the antioxidant and anti-H. pylori activity, the immunomodulatory effect and the antitumor effect in gastric adenocarcinoma cells of the hydroalcoholic extract (SCE) and ethyl acetate (SEAP) and hexane (SHP) partitions from avocado seeds. SEAP obtained better results; therefore, we also performed its chemical profile and the study of its gastroprotective effects in the acute indomethacin-induced gastric ulcer model, through histological analysis and quantification of the biochemical parameters of the inflammation. SEAP and SHP efficiently inhibited gastric tumor cells and H. pylori growth, confirmed by bacterial morphology changes. SEAP presented better results in the capture of ABTS•+, DPPH• , O2 •- , H2O2, HOCl and inhibition of HRP enzyme, besides modulating inflammation by inhibiting IL-6 production significantly. SEAP chromatographic study by ESI FT-ICR MS showed the presence of flavonoids, phenylpropanoids and tannins, such as caffeioylquinic acid, catechin and epicatechin (confirmed by HPLC-DAD) and quercetin and kaempferol derivatives. SEAP reduced gastric lesions characteristics in macroscopic and histological analysis, besides increasing mucus production. In oxidative stress parameters, there was a significant reduction of AOPP and MDA levels with increase of SOD activity. These results show that P. americana Mill. seeds are capable to inhibit the pathways involved in the formation of ulcer and gastric cancer due to the presence of phenolic compounds, being a strategic alternative in the treatment of gastric diseases.
- ItemAvaliação do perfil químico e atividades biológicas de Myrcíaria Strigipes O. Berg (Myrtaceae)(Universidade Federal do Espírito Santo, 2016-03-17) Faitanin, Rafael Destefani; Kitagawa, Rodrigo Rezende; Jamal, Claudia Masrouah; Gonçalves, Rita de Cássia Ribeiro; Silveira, DâmarisBrazil is the owner of the largest and richest genetic diversity of plant species, however, most of them don’t have chemical-biologial studies. Myrciaria strigipes O. Berg (Myrtaceae) is a native species known popularly as "cambucá da praia" and "cabeludinha da praia" and it’s used in traditional medicine for cramps, edema and abdominal pain. The aim of this study was to evaluate the chemical profile, the toxicity and the antimicrobial, antioxidant, thrombolytic activities and α-amylase, α- glucosidase and tyrosinase enzymes inhibition activities of M. strigipes. The chemical profile was characterized from phytochemical screening, High Performance Liquid Chromatography analysis and the fractionation/purification techniques. The toxicity was determined against larvae of Artemia salina Leach. and the antimicrobial activity against Staphylococcus aureus, Escherichia coli and Candida albicans. The antioxidant activity was measured using the synthetic radicals DPPH and ABTS. The in vitro thrombolytic activity was evaluated through the extracts capacity to cause lysis in human blood clot. The inhibitory activities of α-amylase, α-glucosidase and tyrosinase enzymes were determined using in vitro spectrophotometric methods. Preliminary phytochemical analysis of ethanolic extracts of leaves (EEF) and branches (EEG) indicated the presence of flavonoids, coumarins, alkaloids, saponins, tannins, triterpenes, steroids and anthraquinones, yet this was only found in the EEF. They were isolated and/or identified of the EEF five compounds of terpenes class, four pentacyclic triterpenes (friedelin, 28-hydroxyfriedelin, glutinol and ursolic acid) and a tetracyclic steroid (β-sitosterol), and three phenolic compounds (isoquercitrin, hyperoside and ellagic acid). The EEF showed a little toxicity (LD50 = 648.17 µg/mL) and only the EEG showed antimicrobial activity, inhibiting the growth of S. aureus. The EEF and EEG showed antioxidant capacity and promising inhibitory activity on α-glucosidase, EC50 of 40,66 e 18,52 µg/mL, respectively, while the positive control showed EC50 of 37,30 µg/mL. This study reported, for the first time, chemical aspects and biological activities of M. strigipes species, and the results demonstrate the need for more studies to further chemical knowledge and understanding about the mechanism(s) and substance(s) responsible for activities shown by this species.
- ItemEpidemiologia molecular e caracterização da resistência de amostras de Acinetobacter baumannii e Pseudomonas aeruginosa resistentes aos carbapenêmicos provenientes de hospitais da grande Vitória-ES(Universidade Federal do Espírito Santo, 2017-08-30) Vallorini, Thalita Pereira Cabral; Schuenck, Ricardo Pinto; 1º membro da bancaThe emergence and spread of antimicrobial resistance among non-fermenting Gram-negative bacilli, such as Acinetobacter baumannii and Pseudomonas aeruginosa, is a worldwide problem. Carbapenems are beta-lactam antimicrobials indicated for the treatment of serious infections caused by these agents, however, the emergence of multiresistant pathogens seriously threatens the use of this class of drug in the hospital environment. The present work aimed to characterize clinical samples of P. aeruginosa and A. baumannii resistant to cabapenems, as well as the presence of beta-lactam resistance genes, epidemiological profile and susceptibility to antimicrobials used in clinical routine. The antimicrobial susceptibility was performed by disc diffusion and antimicrobial gradient tests. PCR was used to the detection of beta-lactamases-encoding genes and the genetic polymorphism was analyzed by pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) techniques. The species included in this study were: A. baumannii (n = 26) and P. aeruginosa (n = 15). The most of the isolates presented a multiresistance profile to the tested antimicrobials, highlighting the resistance to colistin in nine isolates of A. baumannii. Among the A. baumannii isolates, the prevalent carbapenemase gene (92.3%) was blaOXA-23 and in P. aeruginosa isolates the blaVIM gene was prevalent (33.3%). The PFGE analysis showed the prevalence of two pulsotypes among the A. baumannii: abA (34.6%) and abB (23%), while the P. aeruginosa isolates presented distinct pulsotypes, demonstrating the polyclonal origin of isolates. MLST was performed in five isolates of P. aeruginosa presenting different pulsotypes and STs 357, 2321, 1121, 244 and 227 were found, with two clonal complexes of world importance: CC235 and CC244. In addition, ST357, ST2321 and ST1121 were described for the first time in Brazil.
- ItemEpidemiologia, caracterização microbiológica e avaliação dos custos com o tratamento de infecções do pé diabético(Universidade Federal do Espírito Santo, 2018-03-08) Batista, Julianne Soares Jardim Lacerda; Schuenck, Ricardo Pinto; Spano, Liliana Cruz; Gonçalves, Rita de Cássia RibeiroDiabetic foot infection (DFI) is one of the most serious and costly complications of diabetic patients. This study aimed to analyze the microbiological and epidemiological aspects of DFI and to determine the costs of treating these patients. We collected clinical and epidemiological data from the medical records. To analyze the costs, we collected data referring to the real cost invested in the treatment of each patient. The antimicrobial susceptibility was determined using agar diffusion or E-test. Genetic polymorphisms were analyzed by the PFGE technique. The study included 46 patients with DFI; the majority were men (58.7%) who were elderly (mean age 63.9 ± 10.8 years) with associated cardiovascular diseases (60.8%). The amputation rate was 56.5 %. The average treatment cost of DFI was US$ 9,936.84 (ranging from US$ 14.19 to US$ 58,230.71). Forty-seven isolates were collected, and a predominance of Gram-negative microorganisms (70.8 %) was observed. PFGE revealed a large clonal diversity. Time and costs of hospitalization and antimicrobial treatment were higher in cases of multidrug-resistant and polymicrobial infections. We observed that DFIs are serious complications with long times of hospitalization and costly antimicrobial treatment. Bacterial resistance and polymicrobial nature of wounds can increase the costs of such morbidity.
- ItemEstudo de polimorfismos da região 3' não traduzida do gene HLA-G em pacientes portadores de hepatite B(Universidade Federal do Espírito Santo, 2016-10-07) Fontes, André Martins; Farro, Ana Paula Cazerta; Pancoto, João AlexandreTrés; Guimarães do Bem, Daniela Amorim Melgaço; Palomino, Gustavo MartelliThe World Health Organization (WHO) estimates that about 2 billion people worldwide have had contact with the hepatitis B virus (HBV), and 240 million to become chronic carriers. Hepatitis B is a type of disease in which the pathogenetic mechanisms are not yet fully understood, and it can evolve into chronic form and promote clinical manifestations such as cirrhosis and hepatocellular carcinoma, leading to a large number of deaths annually. Considering the changes in the immune system promoted by HBV, we use a data processing through geographic information system (GIS) technology and epidemiological analysis, in the cities of the northern region of the Espírito Santo State, with the initial objective to characterize the number of individuals infected with HBV. In addition, we used this initial analysis as a basis for evaluation of gene polymorphisms that may be associated with the pathogenesis of the disease, including situated genes within the MHC as the 3 'untranslated region of the HLA-G (presence or absence of 14 pairs base, + 3001 C/T, + 3003 C/T, + 3010C/G, + 3027 A/C, + 3035 C/T, + 3142 C/G, + 3187 A/G e + 3196 C/T). A GIS and epidemiological analysis showed that São Mateus had the largest number of individuals infected with HBV, followed by the cities of Pedro Canario and Jaguaré. The population aged 20-59 years, female sex, were the most affected by the disease, where the main forms of transmission occurred through the use of injectable drugs and dental care. The main drugs used in the treatment were Tenofovir and Entecavir. Among the polymorphic sites studied in individuals of the northern state Espírito Santo and the state of Mato Grosso, only the frequency allelic +3196G [p <0.0130; (OR = 1.540; 95% CI = 1.097 to 2.162)] and frequency genotypic Del/Ins 14-bp [p <0.0194; (OR = 1.645; 95% CI = 1.101 to 2.458)] were more frequent in patients with hepatitis B compared to controls, which may be related to the encouragement of viral escape and progression of infection. Regarding the genotypes +3003C/T, +3003 TT and +3196C/C, these were more frequent in controls compared to patients and are usually associated as microRNAs target (mirRNA) being related to the degradation of mRNA, resulting in low production of soluble and membrane HLA-G isoforms. Together, considering the results found in our study, it is observed that in the north there are many patients infected with HBV, and most of the female, which may be associated with poor adherence to treatment and due to low socio-economic level, sexual transmission, and dental/medical/surgical failures by the standards of biosecurity procedures. Evaluation of genetic polymorphisms in the population suggests that they have genotypes favorable to high production of HLA-G and thus this immunoregulatory molecule could be influencing the development of the chronic form of hepatitis B and the emergence of possible complications of this disease such as liver cirrhosis and hepatocellular carcinoma.
- ItemQualidade da água no município de São Mateus : análise parasitológica, microbiológica e físico-química(Universidade Federal do Espírito Santo, 2018-02-26) Nunes, Luiz Gustavo de Paiva; Oliveira, Maysa do Vale; Souza, Marco Antônio Andrade de; Gradella, Débora Barreto Teresa; Nogueira, Guilherme BicalhoWater is an essential resource for human life, since 75% of the human body is made up of this element. Considering that the preservation of the quality of this resource is fundamental for the health of the population, the present study sought to evaluate the knowledge of the population about the maintenance of the domestic water reservoirs, besides verifying the quality of the water consumed by the population, mainly the effects of the reservoirs in the physico-chemical, parasitological and microbiological characteristics of water, in the city of São Mateus, northern region of Espírito Santo. The collection of socioeconomic data was done through an interview with the residents. For the microbiological analysis, the substrate technique was used, which allows simultaneous detection and identification of total coliforms and E. coli. A sedimentation method by centrifugation (Blagg method) and a flotation method (Sheather modified) was used for the investigation of parasites. The results were evaluated using descriptive and inferential statistical tools, using Pearson's chi-square test and Poisson's regression analysis. The results showed that all the interviewees were aware of the need to clean the water tank, but in 14.29% (n=10) of the households the cleaning was not performed. Regarding the period of cleaning of the water tank, 61.43% (n=43) of the residents stated that six months was the correct period for their realization, with 55.71% (n=39) of the residents effectively performing the cleaning in that period of time. Through the parasitological analysis of the water, only one hookworm egg was found in a sample from the supply network. The results of the microbiological analysis of the water show that the samples from the domestic reservoirs were more contaminated than those of the public supply network, with a significant difference (p=0.008) between the groups, for contamination by total coliforms. Considering the free residual chlorine content, of the 166 analyzed samples, 94.58% (n=157) had levels below the recommended for disinfection of the water, and in no sample from the domestic reservoir the minimum content was reached. The high degree of microbiological contamination of the two groups may be directly associated to the reduced residual chlorine content of the samples, especially those from the domestic reservoirs. The use of the domestic reservoir had a negative effect on the quality of the water, both in relation to free residual chlorine contents and contamination by total coliforms, which could cause harm to the health of the population.