30 research outputs found
Genetic analysis and virulence factors of clinical isolates of Candida albicans from patients with chronic periodontitis with diabetes mellitus type II
Orientadores: Reginaldo Bruno Gonçalves, Cristiane DuqueTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: Candida spp. sĂŁo leveduras comensais que habitam diferentes sĂtios da cavidade bucal. Em indivĂduos saudĂĄveis, sem alteraçÔes imunolĂłgicas, esses microrganismos nĂŁo causam doença. Entretanto, diante de condiçÔes imunossupressoras, essas leveduras podem se tornar mais virulentas e expressar patogenicidade. EspĂ©cies de Candida apresentam diversos fatores de virulĂȘncia, incluindo mecanismos de adesĂŁo e invasĂŁo celular associado Ă produção de enzimas que auxiliam na degradação tecidual e facilitam sua proliferação na mucosa bucal. Estudos tĂȘm demonstrado a presença de Candida sp. em sĂtios periodontais de pacientes com periodontite crĂŽnica, principalmente quando estes sĂŁo imunologicamente comprometidos. Entretanto, ainda Ă© desconhecido o papel desses microrganismos na patogĂȘnese da doença periodontal. Os objetivos do presente trabalho foram: 1) identificar a presença de espĂ©cies de Candida e periodontopatĂłgenos por PCR em sĂtios bucais de pacientes diabĂ©ticos ou nĂŁo com periodontite crĂŽnica; 2) isolar cepas de Candida albicans desses pacientes e avaliĂĄ-las quanto Ă atividade das enzimas proteinase, fosfolipase e hemolisina e os graus de hidrofobicidade da superfĂcie celular, sob diferentes condiçÔes atmosfĂ©ricas, alĂ©m de realizar a anĂĄlise genotĂpica desses isolados; 3) avaliar a capacidade de adesĂŁo e invasĂŁo de cepas de Candida albicans com diferentes graus de hidrofobicidade, em fibroblastos gengivais humanos. As reaçÔes de PCR mostraram que os diabĂ©ticos tiveram maior prevalĂȘncia de Candida spp. principalmente C. albicans e C. dubliniensis, e menor freqĂŒĂȘncia de Tannerella forsythia, quando comparado aos pacientes nĂŁo diabĂ©ticos, para bolsa periodontal e furcas. C. glabrata e C. tropicalis nĂŁo foram encontradas em sĂtios periodontais de pacientes nĂŁo diabĂ©ticos. Dos pacientes diabĂ©ticos, foram isoladas 128 cepas de C. albicans, das quais 51.6% foram determinadas como genĂłtipo B e 48.4% como genĂłtipo A. As condiçÔes ambientais consideradas neste estudo, nĂveis reduzidos de oxigĂȘnio ou anaerobiose, nĂŁo modificaram o tipo de hemĂłlise realizado pelo microrganismo, sendo que a maioria das cepas foi alfa-hemolĂtica. Nesses ensaios, 100% das cepas em anaerobiose apresentaram as colĂŽnias rugosas, enquanto que em ambiente de oxigĂȘnio reduzido, houve variação em relação Ă morfologia e a maioria delas apresentou colĂŽnia lisa. Com relação Ă atividade de proteinase e fosfolipase, cepas de C. albicans nĂŁo produziram as enzimas na ausĂȘncia total de oxigĂȘnio. Em ambiente com nĂvel reduzido de oxigĂȘnio, a maioria das cepas de C. albicans foram fortemente produtoras de proteinase e a maioria das cepas foi positiva para fosfolipase. A hidrofobicidade foi mais alta na condição de anaerobiose. A partir desses resultados, foram selecionadas 16 cepas com alta ou baixa hidrofobicidade e avaliadas quanto Ă capacidade de adesĂŁo e invasĂŁo em fibroblastos gengivais humanos. Foi verificado que ambos os processos foram maiores nas cepas com alta hidrofobicidade. A produção de Ăłxido nĂtrico foi maior para as cepas mais hidrofĂłbicas. Os resultados demonstraram que as espĂ©cies de Candida podem ser encontradas, em grande proporção, em bolsas periodontais e furcas de pacientes portadores de periodontite crĂŽnica, principalmente naqueles acometidos por diabetes mellitus. A maioria das cepas de C. albicans apresentou atividade enzimĂĄtica, que atuaria diretamente na degradação tecidual. AlĂ©m disso, a hidrofobicidade das cepas de C. albicans mostrou estar relacionada Ă maior capacidade de adesĂŁo e invasĂŁo em fibroblastos. Todos esses fatores de virulĂȘncia aumentam a patogenicidade da Candida, que poderia colaborar na progressĂŁo da doença periodontal, principalmente em pacientes imunodeficientesAbstract: Candida spp. are commensal yeasts that inhabit different sites of the oral cavity. In healthy subjects, without immunological alterations, these microorganisms do not cause disease. However, in immunosuppressive conditions, these yeasts can become more virulent and express pathogenicity. Candida species have different virulence factors, including mechanisms of cell adhesion and invasion associated with the production of enzymes that facilitate tissue degradation and their proliferation in oral mucosa. Studies have shown the presence of Candida spp. in periodontal sites of patients with chronic periodontitis, especially when they are immunologically compromised. However is still unknown their role in the pathogenesis of periodontal disease. The objectives of this study were: 1) to identify the presence of Candida species and putative periodontopathogens by PCR in periodontal sites of diabetic or non-diabetic patients with chronic periodontitis; 2) to isolate strains of Candida albicans in these patients and evaluate the proteinase, phospholipase and haemolysin activities and degrees of cell surface hydrophobicity under different atmospheric conditions, besides to performe the genotypic analysis of these isolates; 3) to evaluate the ability of adhesion and invasion of Candida albicans strains with different degrees of hydrophobicity, in human gingival fibroblasts. The PCR reactions revealed that diabetics had higher prevalence of Candida spp., mainly C. albicans and C. dubliniensis, and lower T. forsythia frequency, when compared to non-diabetic patients, for both periodontal sites. C. glabrata and C. tropicalis were not found in periodontal pockets and furcation sites of non-diabetic patients. From diabetic patients, it was isolated 128 strains of C. albicans and 51.6% were determined as genotype B and 48.4% as genotype A. The atmospheric conditions, reduced oxygen and anaerobiosis, did not change the type of hemolysis, and the most of strains were alpha-hemolytic. From these assays, 100% of the strains under anaerobiosis showed rough colonies, whereas in an environment with reduced oxyen was no change in relation to morphology and most of them had smooth colony. Considering proteinase and phospholipase activities, C. albicans strains did not produce the enzymes in the total absence of oxygen. In reduced oxygen, the majority of C. albicans strains were strong proteinase producers and most strains were positive for phospholipase. Hydrophobicity was higher in anaerobic condition. From these results, 16 hydrophobic or hydrophilic strains were selected and evaluated their ability of adhesion and invasion in human gingival fibroblasts. Both processes were greater in strains with high hydrophobicity. The production of nitric oxide was higher for hydrophobic strains. The results showed that Candida species can be found in large proportion, in periodontal pockets and furcation of patients with chronic periodontitis, especially diabetics. The most of C. albicans strains showed enzymatic activity, which could act directly on tissue degradation. Moreover, the hydrophobicity of C. albicans seems to be related to higher capacity of adhesion and invasion in fibroblasts. All these virulence factors enhance the pathogenicity of Candida that could collaborate for the progression of periodontal diseaseDoutoradoMicrobiologia e ImunologiaDoutor em Biologia Buco-Denta
Alternative low doses and routes of administering a prostaglandin F2α analogue to induce luteolysis in Nelore cows
The present study was conducted in order to verify the efficacy of lower doses and alternative routes of a prostaglandin F2α analogue, luprostiol (PGF), for the induction of luteolysis and the precipitation of estrus in nonlactating Nelore cows (Bos taurus indicus). A conventional dose (15 mg) of PGF was compared to doses lower than the conventional dose, which ranges from 10 to 50%, that were administered intramuscularly (IM), intravulvosubmucosally (IVSM), or in the Bai-hui acupuncture site located within the lumbosacral area. The cows were administered PGF 8 day after estrus in the presence of a corpus luteum, and randomly assigned to the following groups: G1 (positive control), 15 mg, IM (n = 23); G2, 7.5 mg, IM (n = 23); G3, 3.75 mg, IM (n = 24); G4, 7.5 mg, IVSM (n = 25); G5, 3.75 mg, Bai-hui acupoint (n = 24); and G6, 1.5 mg, Bai-hui acupoint (n = 25). The results indicated that 50% of a conventional dose of PGF (7.5 mg) resulted in a complete luteal regression (plasma progesterone <1 ng/ml) at Hour 48, and hastened estrus, regardless of whether or not PGF was administered IM or IVSM. Comparatively, 10 or 25% of the conventional dose, even when administered to the Bai-hui acupoint, resulted in an initial reduction in the concentration of progesterone at Hour 24, followed by an increase observed at Hour 48. In conclusion, 25% of a conventional PGF dose administered via the Bai-hui acupoint proved inadequate to induce a complete luteal regression, whereas 50% of a conventional dose administered IM or IVSM was found to be the minimal dose required to induce effectively a complete luteal regression, and to precipitate the onset of estrus in nonlactating Nelore cows
Environmental isolation, biochemical identification, and antifungal drug susceptibility of Cryptococcus species
Novas tendĂȘncias terapĂȘuticas de enfermagem: terapias naturais - programa de atendimento
O presente estudo relata a experiĂȘncia vivenciada pelos enfermeiros na ClĂnica de Enfermagem do Campus UniversitĂĄrio do Grande ABC (UniABC), utilizando as terapias naturais: balanceamento muscular, acupuntura e reflexologia como açÔes terapĂȘuticas de enfermagem. Conclui ressaltando a relevĂąncia da aplicação tĂ©cnico-cientĂfica no desenvolvimento das atividades de enfermagem e a sua conseqĂŒente repercussĂŁo no cuidado de enfermagem
Alternative animal and non-animal models for drug discovery and development: bonus or burden?
FAPESP - FUNDAĂĂO DE AMPARO Ă PESQUISA DO ESTADO DE SĂO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTĂFICO E TECNOLĂGICOMammalian models have served as a basis for R&D over the past decades. Nevertheless, these models are expensive, laborious, may yield results that cannot always be translated into the human in vivo situation and, more recently, have reverberated great social and ethical dilemmas. Hence, the prospect of changes in the global scientific scenario and the Three Rs principle (Reduction, Replacement and Refinement) have encouraged the development of alternative methods to the use of mammals. Despite the efforts, suitable alternative tests are not available in all areas of biomedical research, as regulatory acceptance requires time, prior validation and robust financial and scientific investment. In this perspective, we aim to shed light on the concepts, challenges and perspectives for implementation of innovative alternative animal and non-animal methods in scientific research. The applicability and meaningfulness of invertebrate animal models, in silico analysis and reverse pharmacology are discussed, among other aspects of relevance in today's scenario. Overall, the use of alternative models, including Artemia salina (brine shrimp), Caenorhabditis elegans (roundworm), Danio rerio (zebra fish), Drosophila melanogaster (fruit fly), Galleria mellonella (greater waxmoth) and in silico modelling, increased 909% from 1990 to 2015, as compared to 154% of conventional mammals in the same period. Thus, technological and scientific advancements in the fields of toxicology and drug development seem to have diminished the need for mammalian models. Today, however, mammals still remain critically indispensable to provide - in most cases -reliable data subsidizing and validating translation into the clinical setting.Mammalian models have served as a basis for R&D over the past decades. Nevertheless, these models are expensive, laborious, may yield results that cannot always be translated into the human in vivo situation and, more recently, have reverberated great social and ethical dilemmas. Hence, the prospect of changes in the global scientific scenario and the Three Rs principle (Reduction, Replacement and Refinement) have encouraged the development of alternative methods to the use of mammals. Despite the efforts, suitable alternative tests are not available in all areas of biomedical research, as regulatory acceptance requires time, prior validation and robust financial and scientific investment. In this perspective, we aim to shed light on the concepts, challenges and perspectives for implementation of innovative alternative animal and non-animal methods in scientific research. The applicability and meaningfulness of invertebrate animal models, in silico analysis and reverse pharmacology are discussed, among other aspects of relevance in today's scenario. Overall, the use of alternative models, including Artemia salina (brine shrimp), Caenorhabditis elegans (roundworm), Danio rerio (zebra fish), Drosophila melanogaster (fruit fly), Galleria mellonella (greater waxmoth) and in silico modelling, increased 909% from 1990 to 2015, as compared to 154% of conventional mammals in the same period. Thus, technological and scientific advancements in the fields of toxicology and drug development seem to have diminished the need for mammalian models. Today, however, mammals still remain critically indispensable to provide - in most cases -reliable data subsidizing and validating translation into the clinical setting344681686FAPESP - FUNDAĂĂO DE AMPARO Ă PESQUISA DO ESTADO DE SĂO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTĂFICO E TECNOLĂGICOFAPESP - FUNDAĂĂO DE AMPARO Ă PESQUISA DO ESTADO DE SĂO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTĂFICO E TECNOLĂGICO2011/15984-0; 2013/25080-7310522/2015-
REVISĂO BIBLIOGRĂFICA DO COMPLEXO HIPERPLASIA ENDOMETRIAL CĂSTICA â PIOMETRA EM CADELAS.
O complexo hiperplasia endometrial cĂstica ou piometra Ă© uma doença que acomete grande quantidade de cadelas na clinica veterinĂĄria. Ă uma doença de alta morbidade e mortalidade principalmente em animais de meia idade em fase de diestro. As cadelas com esta enfermidade geralmente encontram-se com apatia, anorexia, e eventualmente sinais mais graves por insuficiĂȘncia renal, como vĂŽmitos diarrĂ©ia, poliĂșria e polidipsia devido a lesĂ”es no parĂȘnquima renal por imunocomplexos previamente formados. A piometra pode se apresentar de duas formas. Nas piometras de cĂ©rvix aberta o corrimento vaginal purulento Ă© comum, e nas de cĂ©rvix fechada o diagnĂłstico Ă© facilitado pela palpação abdominal, por exames laboratoriais, RX, e ultrassonografia. Se nĂŁo tratada pode levar a insuficiĂȘncia renal, septse seguida de choque sĂ©ptico e morte. O tratamento pode ser mĂ©dico ou cirĂșrgico, e a escolha varia de acordo com a apresentação clĂnica da paciente
COMPLEXO HIPERPLASIA ENDOMETRIAL CĂSTICA (PIOMETRA) EM CADELAS â DIAGNĂSTICO E TERAPĂUTICA
Cystic endometrial hyperplasia-pyometra complex is a disease that affects a large number of bitches. It has high morbity and mortality routes, and it occurs particularly in middle-aged bitches during diestrus phase. The bitches with pyometra exhibit apathy, anorexia and possible signs of acute renal failure due to renal parenchyma injuries. There are two types of pyometra: open and closed. The most common sign of open pyometra is the purulent vaginal discharge. In closed pyometra, diagnostic is facilitated by abdominal palpation, laboratory test, ultrasound and X-ray. If untreated, pyometra can cause renal failure, sepsis, septic shock and death. Treatment of pyometra is based on medical or surgical procedures. The treatment choice may vary according to the clinical signs and the state of the patient
New antimicrobial therapies used against fungi present in subgingival sites-A brief review
Although the main reservoir of Candida spp. is believed to be the buccal mucosa, these microorganisms can coaggregate with bacteria in subgingival biofilm and adhere to epithelial cells. The treatment of periodontal disease includes scaling and root planning (SR?) associated with proper oral hygiene. However, some patients may have negative responses to different therapeutic procedures, with a continuous loss of insertion, so the use of antimicrobials is needed as an adjuvant to SRP treatment. The use of a broad-spectrum antibiotic, such as tetracycline and metronidazole, as an aid in periodontal treatment has also been a factor for the development of superinfections by resistant bacteria and Candida species, even in patients with HIV. In the dental practice, the most commonly used antifungals are nystatin and fluconazole. However, the introduction of new drugs like the next generation of azoles is essential before the onset of emergent species in periodontal disease. Plants are good options for obtaining a wide variety of drugs. This alternative could benefit a large population that uses plants as a first treatment option. Plants have been used in medicine for a long time and are extensively used in folk medicine, because they represent an economic alternative, are easily accessible and are applicable to various diseases. Herein, we briefly review the literature pertaining the presence of Candida sp. in periodontal pockets, the conventional antifungal resistance and new therapies that include natural antifungal agents are reviewed. (C) 2011 Elsevier Ltd. All rights reserved
In silico approaches for screening molecular targets in Candida albicans: A proteomic insight into drug discovery and development
Candida species are opportunistic pathogens which can cause conditions ranging from simple mucocutaneous infections to fungemia and death in immunosuppressed and hospitalized patients. Candida albicans is considered to be the species mostly associated with fungal infections in humans and, therefore, the mostly studied yeast. This microorganism has survival and virulence factors which, allied to a decreased host immunity response, make infection more difficult to control. Today, the current limited antifungal arsenal and a dramatic increase in fungal resistance have driven the need for the synthesis of drugs with novel mechanisms of action. However, the development of a new drug from discovery to marketing takes a long time and is highly costly. The objective of this review is to show that with advances in biotechnology and biofinformatics, in silico tools such as molecular docking can optimize such a timeline and reduce costs, while contributing to the design and development of targeted drugs. Here we highlight the most promising protein targets in Candida albicans for the development of drugs with new mechanisms of action8426469CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTĂFICO E TECNOLĂGICO - CNPQ141129/2017-
Unexplored endemic fruit species from Brazil: antibiofilm properties, insights into mode of action, and systemic toxicity of four Eugenia spp
CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTĂFICO E TECNOLĂGICOFAPESP - FUNDAĂĂO DE AMPARO Ă PESQUISA DO ESTADO DE SĂO PAULOBrazilian endemic fruit species have aroused attention due to their highly valuable, yet unexplored, agroindustrial, food and therapeutic potential. Herein, we describe the antifungal activity of four Eugenia spp. against Candida albicans biofilms, and further demonstrate insights into their potential mode(s) of action and toxicity in vitro and in vivo. Extracts from different parts (seeds, pulps, leaves) of E. leitonii (EL), E. brasiliensis (EB), E. myrcianthes (EM) and E. involucrata (EI) were obtained (S23 degrees 23',W45 degrees 39') and chemically characterized by GC/MS. The active extracts were tested against C albicans biofilm viability and architecture, as well as mode of action, and toxicology using RAW 264.7 macrophages and Galleria mellonella larvae. The MIC values ranged from 15.62 to > 2000 mu g/mL. The most active extracts were EL (seed, 15.62 mu g/mL) and EB (leaf and seeds, 31.25 and 15.62 mu g/mL, respectively). Treatment with these extracts at 10xMIC reduced biofilm viability by 54-55% (P 0.05) and G. mellonella larvae, with mean in vivo LD50 of 1500 mg/kg (EL, seeds); 2500 mg/kg (EB, seeds); and 1250 mg/kg (EB, leaf). The phenolic compounds epicatechin and gallic acid were the major constituents in the extracts. Our findings may open avenues for the application of these yet unexplored native fruits in the food and pharmaceutical industry. (C) 2017 Elsevier Ltd. All rights reserved.Brazilian endemic fruit species have aroused attention due to their highly valuable, yet unexplored, agroindustrial, food and therapeutic potential. Herein, we describe the antifungal activity of four Eugenia spp. against Candida albicans biofilms, and further demonstrate insights into their potential mode(s) of action and toxicity in vitro and in vivo. Extracts from different parts (seeds, pulps, leaves) of E. leitonii (EL), E. brasiliensis (EB), E. myrcianthes (EM) and E. involucrata (EI) were obtained (S23 degrees 23',W45 degrees 39') and chemically characterized by GC/MS. The active extracts were tested against C albicans biofilm viability and architecture, as well as mode of action, and toxicology using RAW 264.7 macrophages and Galleria mellonella larvae. The MIC values ranged from 15.62 to > 2000 mu g/mL. The most active extracts were EL (seed, 15.62 mu g/mL) and EB (leaf and seeds, 31.25 and 15.62 mu g/mL, respectively). Treatment with these extracts at 10xMIC reduced biofilm viability by 54-55% (P 0.05) and G. mellonella larvae, with mean in vivo LD50 of 1500 mg/kg (EL, seeds); 2500 mg/kg (EB, seeds); and 1250 mg/kg (EB, leaf). The phenolic compounds epicatechin and gallic acid were the major constituents in the extracts. Our findings may open avenues for the application of these yet unexplored native fruits in the food and pharmaceutical industry105280287CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTĂFICO E TECNOLĂGICOFAPESP - FUNDAĂĂO DE AMPARO Ă PESQUISA DO ESTADO DE SĂO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTĂFICO E TECNOLĂGICOFAPESP - FUNDAĂĂO DE AMPARO Ă PESQUISA DO ESTADO DE SĂO PAULO474335/201352011/15984-0; 2011/25080-7; 2013/26251-