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1.
BMC Infect Dis ; 20(1): 6, 2020 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-31900118

RESUMO

BACKGROUND: An efficient surface cleaning strategy would first target cleaning to surfaces that make large contributions to the risk of infections. METHODS: In this study, we used data from the literature about methicillin-resistant Staphylococcus aureus (MRSA) and developed an ordinary differential equations based mathematical model to quantify the impact of contact heterogeneity on MRSA transmission in a hypothetical 6-bed intensive care unit (ICU). The susceptible patients are divided into two types, these who are cared by the same nurse as the MRSA infected patient (Type 1) and these who are not (Type 2). RESULTS: The results showed that the mean MRSA concentration on three kinds of susceptible patient nearby surfaces was significantly linearly associated with the hand-touch frequency (p < 0.05). The noncompliance of daily cleaning on patient nearby high-touch surfaces (HTSs) had the most impact on MRSA transmission. If the HTSs were not cleaned, the MRSA exposure to Type 1 and 2 susceptible patients would increase 118.4% (standard deviation (SD): 33.0%) and 115.4% (SD: 30.5%) respectively. The communal surfaces (CSs) had the least impact, if CSs were not cleaned, the MRSA exposure to Type 1 susceptible patient would only increase 1.7% (SD: 1.3). The impact of clinical equipment (CE) differed largely for two types of susceptible patients. If the CE was not cleaned, the exposure to Type 1 patients would only increase 8.4% (SD: 3.0%), while for Type 2 patients, it can increase 70.4% (SD: 25.4%). CONCLUSIONS: This study provided a framework to study the pathogen concentration dynamics on environmental surfaces and quantitatively showed the importance of cleaning patient nearby HTSs on controlling the nosocomial infection transmission via contact route.


Assuntos
Busca de Comunicante/métodos , Unidades de Terapia Intensiva , Staphylococcus aureus Resistente à Meticilina/fisiologia , Modelos Teóricos , Infecções Estafilocócicas/transmissão , Busca de Comunicante/estatística & dados numéricos , Infecção Hospitalar/transmissão , Detergentes/farmacologia , Desinfecção , Feminino , Higiene das Mãos/estatística & dados numéricos , Humanos , Controle de Infecções/métodos , Transmissão de Doença Infecciosa do Paciente para o Profissional/estatística & dados numéricos , Transmissão de Doença Infecciosa do Profissional para o Paciente/estatística & dados numéricos , Unidades de Terapia Intensiva/estatística & dados numéricos , Masculino , Resistência a Meticilina/fisiologia , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Staphylococcus aureus Resistente à Meticilina/patogenicidade , Propriedades de Superfície
2.
J Appl Microbiol ; 128(5): 1324-1338, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31872930

RESUMO

AIMS: To develop a method that is able to determine the microbial reduction in different dishwasher cleaning cycles and differentiate between different program parameters used. METHODS AND RESULTS: Stainless steel biomonitors were contaminated with Micrococcus luteus or Entereococcus faecium and cleaned in a specially programmed household dishwasher with different cleaning temperatures and durations. No detergent, bleach-free detergent or detergent containing activated oxygen bleach was used. The logarithmic reduction (LR) was determined. The microbial reduction depended on the cleaning temperature, the duration of the cleaning cycles and the detergent type used. LR increased with higher temperatures, longer cleaning cycles and use of detergent. CONCLUSIONS: The factors cleaning cycle temperature, cleaning cycle duration, final rinsing temperature and the use of detergent all contributed to the reduction of test-strains in dishwasher cycles. A combination of longer dishwashing cycles and increased temperatures resulted in LRmax of the microbial load. SIGNIFICANCE AND IMPACT OF THE STUDY: Cycles in domestic appliances are very diverse; therefore a standardized method to determine their ability to reduce the microbial load is of great use. The method described here is able to demonstrate the reductions achieved by dishwashing cycles with different parameters and might help to find the necessary balance between energy saving and an acceptable level of hygiene.


Assuntos
Desinfecção/instrumentação , Desinfecção/métodos , Utensílios Domésticos/normas , Bactérias/efeitos dos fármacos , Bactérias/crescimento & desenvolvimento , Utensílios de Alimentação e Culinária/normas , Detergentes/farmacologia , Desinfecção/normas , Higiene/normas , Aço Inoxidável , Temperatura , Fatores de Tempo
3.
PLoS One ; 14(12): e0225475, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31790434

RESUMO

Rapid sample preparation is one of the leading bottlenecks to low-cost and efficient sample component detection. To overcome this setback, a technology known as Lyse-It has been developed to rapidly (less than 60 seconds) lyse Gram-positive and-negative bacteria alike, while simultaneously fragmenting DNA/RNA and proteins into tunable sizes. This technology has been used with a variety of organisms, but the underlying mechanism behind how the technology actually works to fragment DNA/RNA and proteins has hitherto been studied. It is generally understood how temperature affects cellular lysing, but for DNA/RNA and protein degradation, the temperature and amount of energy introduced by microwave irradiation of the sample, cannot explain the degradation of the biomolecules to the extent that was being observed. Thus, an investigation into the microwave generation of reactive oxygen species, in particular singlet oxygen, hydroxyl radicals, and superoxide anion radicals, was undertaken. Herein, we probe one aspect, the generation of reactive oxygen species (ROS), which is thought to contribute to a non-thermal mechanism behind biomolecule fragmentation with the Lyse-It technology. By utilizing off/on (Photoinduced electron transfer) PET fluorescent-based probes highly specific for reactive oxygen species, it was found that as oxygen concentration in the sample and/or microwave irradiation power increases, more reactive oxygen species are generated and ultimately, more oxidation and biomolecule fragmentation occurs within the microwave cavity.


Assuntos
Métodos Analíticos de Preparação de Amostras/métodos , Técnicas Bacteriológicas/métodos , Fragmentação do DNA/efeitos dos fármacos , Detergentes/farmacologia , Estabilidade de RNA/efeitos dos fármacos , Fragmentação do DNA/efeitos da radiação , DNA Bacteriano/química , DNA Bacteriano/efeitos dos fármacos , DNA Bacteriano/efeitos da radiação , Hidrólise/efeitos da radiação , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/genética , Listeria monocytogenes/efeitos da radiação , Micro-Ondas , Oxirredução/efeitos dos fármacos , Oxirredução/efeitos da radiação , Oxigênio/análise , Oxigênio/metabolismo , Proteólise/efeitos dos fármacos , Proteólise/efeitos da radiação , Estabilidade de RNA/efeitos da radiação , RNA Bacteriano/química , RNA Bacteriano/efeitos dos fármacos , RNA Bacteriano/efeitos da radiação , Espécies Reativas de Oxigênio/análise , Espécies Reativas de Oxigênio/metabolismo , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/genética , Staphylococcus aureus/efeitos da radiação , Temperatura , Fatores de Tempo , Vibrio cholerae/efeitos dos fármacos , Vibrio cholerae/genética , Vibrio cholerae/efeitos da radiação
4.
Rev Lat Am Enfermagem ; 27: e3211, 2019.
Artigo em Inglês, Português, Espanhol | MEDLINE | ID: mdl-31826156

RESUMO

OBJECTIVE: to evaluate the potential contamination of enzymatic detergent from its reuse and to identify the microbiological profile in the solution used to clean gastrointestinal endoscopic devices. METHOD: cross-sectional study based on microbiological analysis of 76 aliquots of 19 different enzymatic detergent solutions used to clean endoscopic devices. The aliquots were homogenized, subjected to Millipore® 0.45 µm membrane filtration and the presumptive identification of microorganisms was performed by biochemical-physiological methods according to previously established specific bacterial groups that are of clinical and epidemiological relevance. RESULTS: the mean values, as well as the standard deviation and the median, of the enzymatic detergent microbial load increased as the solution was reused. There was a significant difference between the means of after first use and after fifth reuse. A total of 97 microorganisms were identified, with predominance of the coagulase-negative Staphylococcus, Pseudomonas spp., Klebsiella spp., Enterobacter spp. genus, and Escherichia coli species. CONCLUSION: the reuse of the enzymatic detergent solution is a risk to the safe processing of endoscopic devices, evidenced by its contamination with pathogenic potential microorganisms, since the enzymatic detergent has no bactericidal property and can contribute as an important source for outbreaks in patients under such procedures.


Assuntos
Detergentes/efeitos adversos , Contaminação de Equipamentos , Gastroscópios/efeitos adversos , Gastroscópios/microbiologia , Carga Bacteriana , Estudos Transversais , Detergentes/farmacologia , Transmissão de Doença Infecciosa , Bactérias Gram-Negativas/classificação , Bactérias Gram-Negativas/isolamento & purificação , Bactérias Gram-Positivas/classificação , Bactérias Gram-Positivas/isolamento & purificação , Humanos , Controle de Infecções
5.
Int J Mycobacteriol ; 8(4): 390-396, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31793511

RESUMO

Background: Ubiquitous presence of Mycobacterium fortuitum and ability to cause infections in human beings, hints toward its integral resistance against environmental and host stress conditions. With an aim to identify genes responsible for adapting in vitro acidic stress of M. fortuitum, in the previous study, TnphoA random mutagenesis identified acid susceptible mutant MT727, with mutation in ribosomal maturation factor encoding gene rimP, to be mutated. The present study was conducted to explore virulent behavior as well as growth behavior under in vitro stress conditions. Methods: Acid susceptible transposon mutant MT727 was injected intravenously in female BALB/c mice and kidney tissue was analyzed for the bacillary load as well as pathological characterization. Cytokine profiling of MT727-infected mice serum was done. MT727 was also subjected to various in vitro stress conditions, including detergent stress, heat stress, and hypoxic stress. The viable count of bacteria under different stress conditions was determined at regular time interval. Results: Mutant MT727 showed slight variation in bacillary load in vivo; however, defective growth behavior under detergent and hypoxic stress was observed when compared to wild type strain. Conclusion: Results conclude probable involvement of rimP gene in survival of M. fortuitum under hypoxic stress and detergent stress conditions.


Assuntos
Ácidos/farmacologia , Mutação , Mycobacterium fortuitum/efeitos dos fármacos , Mycobacterium fortuitum/genética , Animais , Proteínas de Bactérias/genética , Citocinas/imunologia , Detergentes/farmacologia , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Dodecilsulfato de Sódio/farmacologia , Estresse Fisiológico
6.
Transfus Apher Sci ; 58(6): 102665, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31740165

RESUMO

BACKGROUND: Therapeutic Plasma Exchange (TPE) is the primary therapy of immune-mediated Thrombotic Thrombocytopenic Purpura (iTTP). Efficacy and safety data for TPE of iTTP have been assessed with Quarantine and Solvent-Detergent inactivated (SD) plasma. Here, amotosalen-UVA pathogen inactivated (AI) plasma, also in routine use, was evaluated in iTTP. METHODS: We conducted a retrospective review of iTTP cases prospectively reported to the French national registry (2010-2013). Cases reviewed underwent TPE with ≥70% of either AI or SD plasma. The primary endpoint was time to platelet count recovery; secondary endpoints were related to follow-up (sustained remission, relapses, flare-ups and refractoriness). RESULTS: 30 Test patients were identified in the AI group which could be timely matched to 40 Control patients in the SD group. The groups were fairly comparable for clinical presentation. Major findings were: (i) iTTP patients were exposed to lower plasma volumes in the AI group than in the SD group; (ii) Recovery rates were comparable between the groups. Median time to platelet count recovery (>150 × 109/L) trended to be shorter in the AI group though non significantly. Tolerance of AI vs SD plasma was of comparable frequency and severity in either group. CONCLUSION: TPE with Amotosalen-inactivated plasma demonstrated therapeutic efficacy and tolerability for iTTP patients. In view of the retrospective design, confirmation of these results is required in larger prospective studies.


Assuntos
Detergentes/farmacologia , Furocumarinas/farmacologia , Plasma/metabolismo , Púrpura Trombocitopênica Trombótica/terapia , Adulto , Estudos de Coortes , Feminino , Furocumarinas/efeitos adversos , Humanos , Masculino , Troca Plasmática , Solventes , Resultado do Tratamento
7.
PLoS One ; 14(10): e0222932, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31618200

RESUMO

The present study mainly consists of a re-evaluation of the rate at which C12E8, a typical non-ionic detergent used for membrane studies, is able to dissociate from biological membranes, with sarcoplasmic reticulum membrane vesicles being used as an example. Utilizing a brominated derivative of C12E8 and now stopped-flow fluorescence instead of rapid filtration, we found that the rate of dissociation of this detergent from these membranes, merely perturbed with non-solubilizing concentrations of detergent, was significantly faster (t1/2 < 10 ms) than what had previously been determined (t1/2 ~300-400 ms) from experiments based on a rapid filtration protocol using 14C-labeled C12E8 and glass fiber filters (Binding of a non-ionic detergent to membranes: flip-flop rate and location on the bilayer, by Marc le Maire, Jesper Møller and Philippe Champeil, Biochemistry (1987) Vol 26, pages 4803-4810). We here pinpoint a methodological problem of the earlier rapid filtration experiments, and we suggest that the true overall dissociation rate of C12E8 is indeed much faster than previously thought. We also exemplify the case of brominated dodecyl-maltoside, whose kinetics for overall binding to and dissociation from membranes comprise both a rapid and a sower phase, the latter being presumably due to flip-flop between the two leaflets of the membrane. Consequently, equilibrium is reached only after a few seconds for DDM. This work thereby emphasizes the interest of using the fluorescence quenching associated with brominated detergents for studying the kinetics of detergent/membrane interactions, namely association, dissociation and flip-flop rates.


Assuntos
Detergentes/farmacologia , Filtração/métodos , Membranas Intracelulares/metabolismo , Detergentes/química , Retículo Sarcoplasmático/metabolismo , Espectrometria de Fluorescência , Vesículas Transportadoras/metabolismo
8.
PLoS One ; 14(9): e0223008, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31568482

RESUMO

Nucleases are enzymes that can degrade genomic DNA and RNA that decrease the accuracy of quantitative measures of those nucleic acids. Here, we study conventional heating, standard microwave irradiation, and Lyse-It, a microwave-based lysing technology, for the potential to fragment and inactivate DNA and RNA endonucleases. Lyse-It employs the use of highly focused microwave irradiation to the sample ultimately fragmenting and inactivating RNase A, RNase B, and DNase I. Nuclease size and fragmentation were determined visually and quantitatively by SDS polyacrylamide gel electrophoresis and the mini-gel Agilent 2100 Bioanalyzer system, with a weighted size calculated to depict the wide range of nuclease fragmentation. Enzyme activity assays were conducted, and the rates were calculated to determine the effect of various lysing conditions on each of the nucleases. The results shown in this paper clearly demonstrate that Lyse-It is a rapid and highly efficient way to degrade and inactivate nucleases so that nucleic acids can be retained for down-stream detection.


Assuntos
Desoxirribonuclease I/química , Fragmentos de Peptídeos/análise , Ribonuclease Pancreático/química , Ribonucleases/química , DNA/química , Desoxirribonuclease I/efeitos dos fármacos , Desoxirribonuclease I/efeitos da radiação , Detergentes/farmacologia , Eletroforese em Gel de Poliacrilamida , Temperatura Alta , Hidrólise , Micro-Ondas , Peso Molecular , Proteólise/efeitos dos fármacos , Proteólise/efeitos da radiação , RNA/química , Ribonuclease Pancreático/efeitos dos fármacos , Ribonuclease Pancreático/efeitos da radiação , Ribonucleases/efeitos dos fármacos , Ribonucleases/efeitos da radiação , Soluções
9.
Int J Biochem Cell Biol ; 116: 105612, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31546020

RESUMO

BACKGROUND/AIMS: Epigallocatechin-3-gallate (EGCG), a major catechin found in green tea, plays an important anti-tumor role and is involved in various other biological processes, such as, neuroprotection by prevention of aggregation of misfolded proteins generated because of genetic defects. Surfactant protein A2 mutations (G231V and F198S) have been identified to be associated with pulmonary fibrosis and lung cancer, and these mutations cause protein aggregation, instability as well as secretion deficiency. The present study focused on investigating the inhibitory effects of EGCG on aggregation of mutant SP-A2 and elucidating the potential mechanisms underlying this action. METHODS: Wild-type and mutant SP-A2 were transiently expressed in CHO-K1 cells. The aggregated and soluble proteins were separated into NP-40-insoluble and NP-40-soluble fractions. Protein stability was validated by chymotrypsin limited proteolysis assay. Western blot and RT-PCR were used to determine the protein and mRNA expression level, respectively. RESULTS: Mutant SP-A2 alone or wild-type SP-A2 co-expressed with G231V formed NP-40-insoluble aggregates in CHO-K1 cells. EGCG significantly suppressed this aggregation and alleviated mutant SP-A2 accumulation in the ER. When combined with 4-PBA, EGCG treatment completely blocked mutant SP-A2 aggregate formation. Though secretion of mutant protein was not affected, EGCG facilitated protein instability in both wild-type and mutant protein. Importantly, MG132, a proteasome inhibitor, reversed EGCG-induced aggregate reduction. CONCLUSIONS: EGCG inhibits aggregation of misfolded SP-A2 via induction of protein instability and activation of proteasomal pathway for aggregate degradation.


Assuntos
Catequina/análogos & derivados , Complexo de Endopeptidases do Proteassoma/efeitos dos fármacos , Agregados Proteicos/efeitos dos fármacos , Proteólise/efeitos dos fármacos , Proteína A Associada a Surfactante Pulmonar/química , Animais , Butilaminas/farmacologia , Células CHO , Catequina/farmacologia , Cricetulus , Inibidores de Cisteína Proteinase/farmacologia , Detergentes/farmacologia , Expressão Gênica , Leupeptinas/farmacologia , Mutação , Octoxinol/farmacologia , Estabilidade Proteica , Fibrose Pulmonar/metabolismo , Proteína A Associada a Surfactante Pulmonar/genética , Proteína A Associada a Surfactante Pulmonar/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Solubilidade
10.
Protein Eng Des Sel ; 32(3): 129-143, 2019 12 13.
Artigo em Inglês | MEDLINE | ID: mdl-31504920

RESUMO

In the accompanying paper, we described evolving a lipase to the point where variants were soluble, stable and capable of degrading C8 TAG and C8 esters. These variants were tested for their ability to survive in an environment that might be encountered in a washing machine. Unfortunately, they were inactivated both by treatment with a protease used in laundry detergents and by very low concentrations of sodium dodecyl sulfate (SDS). In addition, all the variants had very low levels of activity with triglycerides with long aliphatic chains and with naturally occurring oils, like olive oil. Directed evolution was used to select variants with enhanced properties. In the first 10 rounds of evolution, the primary screen was selected for variants capable of hydrolyzing olive oil whereas the secondary screen was selected for enhanced tolerance towards a protease and SDS. In the final six rounds of evolution, the primary and secondary screens identified variants that retained activity after treatment with SDS. Sixteen cycles of evolution gave variants with greatly enhanced lipolytic activity on substrates that had both long (C16 and C18) as well as short (C3 and C8) chains. We found variants that were stable for more than 3 hours in protease concentrations that rapidly degrade the wild-type enzyme. Enhanced tolerance towards SDS was found in variants that could break down naturally occurring lipid and resist protease attack. The amino acid changes that gave enhanced properties were concentrated in the cap domain responsible for substrate binding.


Assuntos
Evolução Molecular Direcionada , Lipase/genética , Lipase/metabolismo , Peptídeo Hidrolases/metabolismo , Engenharia de Proteínas , Triglicerídeos/metabolismo , Detergentes/farmacologia , Estabilidade Enzimática/efeitos dos fármacos , Estabilidade Enzimática/genética , Hidrólise , Lipase/química , Proteólise , Dodecilsulfato de Sódio/farmacologia , Solubilidade , Especificidade por Substrato , Temperatura
11.
Int J Biol Macromol ; 140: 1037-1046, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31449862

RESUMO

Azo dyes are the most widely applied chemical dyes that have also raised great concerns for environmental contamination and human health issues. There has been a growing interest in discovering bioremediation methods to degrade azo dyes for environmental and economic purposes. Azoreductases are key enzymes evolved in nature capable of degrading azo dyes. The current work reports the identification, expression, and properties of a novel azoreductase (AzoRed2) from Streptomyces sp. S27 which shows an excellent stability against pH change and organic solvents. To overcome the requirements of coenzyme while degrading azo dyes, we introduced a coenzyme regeneration enzyme, Bacillus subtilis glucose 1-dehydrogenase (BsGDH), to construct a recycling system in living cells. The whole-cell biocatalyst containing AzoRed2 and BsGDH was used to degrade a representative azo dye methyl red. The degradation rate of methyl red was up to 99% in 120 min with high substrate concentration (250 µM) and no external coenzyme added. The degradation rate was still 98% in the third batch trial. To sum up, a novel azoreductase with good properties was found, which was applied to construct whole-cell biocatalyst. Both the enzymes and whole-cell biocatalysts are good candidates for the industrial wastewater treatment and environmental restoration.


Assuntos
Compostos Azo/isolamento & purificação , NADH NADPH Oxirredutases/metabolismo , Streptomyces/enzimologia , Águas Residuárias/química , Sequência de Aminoácidos , Compostos Azo/química , Biocatálise , Biodegradação Ambiental , Detergentes/farmacologia , Estabilidade Enzimática , Concentração de Íons de Hidrogênio , Íons , Metais/farmacologia , NADH NADPH Oxirredutases/química , Filogenia , Solventes/farmacologia , Espectrofotometria Ultravioleta , Especificidade por Substrato , Temperatura
12.
J Biol Chem ; 294(36): 13218-13223, 2019 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-31362983

RESUMO

A hallmark of G-protein-coupled receptors (GPCRs) is the conversion of external stimuli into specific cellular responses. In this tightly-regulated process, extracellular ligand binding by GPCRs promotes specific conformational changes within the seven transmembrane helices, leading to the coupling and activation of intracellular "transducer" proteins, such as heterotrimeric G proteins. Much of our understanding of the molecular mechanisms that govern GPCR activation is derived from experiments with purified receptors reconstituted in detergent micelles. To elucidate the influence of the phospholipid bilayer on GPCR activation, here we interrogated the functional, pharmacological, and biophysical properties of a GPCR, the ß2-adrenergic receptor (ß2AR), in high-density lipoprotein (HDL) particles. Compared with detergent-reconstituted ß2AR, the ß2AR in HDL particles had greatly enhanced levels of basal (constitutive) activity and displayed increased sensitivity to agonist activation, as assessed by activation of heterotrimeric G protein and allosteric coupling between the ligand-binding and transducer-binding pockets. Using 19F NMR spectroscopy, we directly linked these functional differences in detergent- and HDL-reconstituted ß2AR to a change in the equilibrium between inactive and active receptor states. The contrast between the low levels of ß2AR constitutive activity in cells and the high constitutive activity observed in an isolated phospholipid bilayer indicates that ß2AR basal activity depends on the reconstitution system and further suggests that various cellular mechanisms suppress ß2AR basal activity physiologically. Our findings provide critical additional insights into GPCR activation and reveal how dramatically reconstitution systems can impact membrane protein function.


Assuntos
Detergentes/farmacologia , Fosfolipídeos/metabolismo , Receptores Adrenérgicos beta 2/metabolismo , Humanos
13.
BMC Pharmacol Toxicol ; 20(1): 33, 2019 05 28.
Artigo em Inglês | MEDLINE | ID: mdl-31138331

RESUMO

BACKGROUND: Antibiotic-resistant pathogens are an emerging threat in this century. Epinecidin-1 is a multi-functional Antimicrobial Peptide (AMP) produced by Orange-spotted grouper (Epinephelus coioides) has been shown to have extensive potentials as an alternative for current antibiotics. Due to the huge costs for the study and the production of a new drug, if an antimicrobial peptide has other beneficial functions in addition to antimicrobial activities, it would be preferred. METHODS: In this study, properties and applications of Epinecidin-1 were investigated and addressed comprehensively. To achieve this, the Google Scholar search engine and three databases of PubMed, Scopus, and Web of Science were used. RESULTS: Epinecidin-1 is a cationic AMP with an alpha-helical structure. Seven functional usages of this peptide have been reported in the literature including antibacterial, antifungal, antiviral, antiprotozoal, anticancer, immunomodulatory, and wound healing properties. Moreover, this peptide has high potential to be used as an active ingredient in cleaning solutions as well as application in vaccine production. CONCLUSION: Due to significant antimicrobial activities tested on bacteria such as Staphylococcus aureus and Helicobacter pylori and also wound healing properties, Epi-1 has high potential to be considered as an important candidate for the production of new drugs and treatment of various infections including diabetic foot ulcer and peptic ulcer. Moreover, adjuvant-like properties of Epi-1 make it a suitable candidate for the studies related to an adjuvant. Other attractive properties such as anticancer effects have also been reported for this peptide which encourages further studies on this peptide.


Assuntos
Anti-Infecciosos , Peptídeos Catiônicos Antimicrobianos , Antineoplásicos , Detergentes , Proteínas de Peixes , Fatores Imunológicos , Animais , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Detergentes/química , Detergentes/farmacologia , Proteínas de Peixes/química , Proteínas de Peixes/farmacologia , Humanos , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Estrutura Molecular , Vacinas , Cicatrização/efeitos dos fármacos
14.
J Hosp Infect ; 103(1): e101-e104, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31112729

RESUMO

The use of microfibre cloths with either water, detergent or disinfectant is currently recommended for hospital cleaning. This study explored the efficacy of a microfibre cloth with either water or detergent/disinfectant or sporicidal products using the ASTM2967-15 standard against Staphylococcus aureus, Acinetobacter baumannii and Clostridium difficile spores. The use of detergent/disinfectant or sporicidal products had a significantly (analysis of variance (ANOVA), P<0.001) better activity than water alone in reducing bacteria and spores' viability, and in reducing the transfer microorganisms between surfaces. The use of water alone with a microfibre cloth is less effective and should not replace the use of biocidal products.


Assuntos
Detergentes/farmacologia , Desinfetantes/farmacologia , Desinfecção/métodos , Microbiologia Ambiental , Serviço Hospitalar de Limpeza/métodos , Têxteis , Acinetobacter baumannii/efeitos dos fármacos , Acinetobacter baumannii/isolamento & purificação , Carga Bacteriana , Clostridium difficile/efeitos dos fármacos , Clostridium difficile/isolamento & purificação , Viabilidade Microbiana/efeitos dos fármacos , Esporos Bacterianos/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/isolamento & purificação
15.
Biochem Soc Trans ; 47(3): 919-932, 2019 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-31085615

RESUMO

Biological membranes form the boundaries to cells. They are integral to cellular function, retaining the valuable components inside and preventing access of unwanted molecules. Many different classes of molecules demonstrate disruptive properties to the plasma membrane. These include alcohols, detergents and antimicrobial agents. Understanding this disruption and the mechanisms by which it can be mitigated is vital for improved therapeutics as well as enhanced industrial processes where the compounds produced can be toxic to the membrane. This mini-review describes the most common molecules that disrupt cell membranes along with a range of in vitro liposome-based techniques that can be used to monitor and delineate these disruptive processes.


Assuntos
Lipossomos , Modelos Biológicos , Anestésicos Locais/farmacologia , Anti-Infecciosos/farmacologia , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Detergentes/farmacologia , Solventes/farmacologia
16.
Int J Biol Macromol ; 131: 989-997, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30917914

RESUMO

Lipase B from Candida antarctica (CALB), lipase from Rhizomucor miehei (RML) and phospholipase Lecitase Ultra (LEU) were immobilized via interfacial activation and their stabilities were compared. Immobilized CALB was much more stable than immobilized RML or LEU. That meant that, if they were coimmobilized, after the inactivation of the least stable lipases, CALB should be discarded even though it may maintain full activity. This could be solved by sequential coimmobilization on octyl-glyoxyl (OCGLX). First, CALB was immobilized on OCGLX getting some covalent bonds between most of the CALB molecules and the support. Then, after reduction of CALB immobilized on OCGLX, RML or LEU can be immobilized on the support via interfacial activation. These enzymes could be released from the support just by using detergents, without affecting CALB activity. After optimization of the lipase desorption conditions, the bi-combilipases CALB/RML and CALB/LEU or the triple-combilipase CALB/RML/LEU could be submitted to several cycles of immobilized biocatalyst inactivation, desorption and enzyme reloading keeping the activity of the immobilized CALB almost intact. This way, by using OCGLX and a stepwise immobilization protocol, discarding all coimmobilized lipases when one becomes inactivated is no longer required. Thus, the most stable ones can be reused in several cycles.


Assuntos
Enzimas Imobilizadas , Glioxilatos/química , Lipase/química , Sefarose/química , Biocatálise , Candida/enzimologia , Detergentes/farmacologia , Ativação Enzimática/efeitos dos fármacos , Estabilidade Enzimática , Proteínas Fúngicas/química , Cinética
17.
Viruses ; 11(3)2019 03 07.
Artigo em Inglês | MEDLINE | ID: mdl-30866548

RESUMO

BACKGROUND: Chikungunya virus (CHIKV) and Mayaro virus (MAYV) are closely related members of the Semliki Forest complex within the genus alphavirus and are transmitted by arthropods, causing acute febrile illness in humans. CHIKV has spread to almost all continents, whereas autochthonous MAYV infections have been reported in South America and in the Caribbean. Nevertheless, there was concern about potential spread of MAYV to other regions similar to CHIKV in the past. The risk for transmission of emerging viruses by blood transfusion and the safety of plasma-derived medicinal products (PDMPs) are constant concerns. The manufacturing processes of PDMPs include procedures to inactivate/remove viruses. METHODS: In this study, we investigated the reduction of MAYV and CHIKV by heat inactivation in various matrices, solvent/detergent treatment and nanofiltration. RESULTS: Unexpectedly, MAYV was significantly more resistant to heat and solvent/detergent treatment compared to CHIKV. However, being similar in size, both MAYV and CHIKV were removed below the detection limit by 35 nm virus filters. CONCLUSIONS: The inactivation profiles of different alphavirus members vary considerably, even within the Semliki Forest Complex. However, robust dedicated viral inactivation/removal procedures commonly used in the plasma product industry are effective in inactivating or removing MAYV and CHIKV.


Assuntos
Alphavirus/isolamento & purificação , Febre de Chikungunya/prevenção & controle , Vírus Chikungunya/isolamento & purificação , Temperatura Alta , Plasma/virologia , Inativação de Vírus , Animais , Febre de Chikungunya/transmissão , Chlorocebus aethiops , Detergentes/farmacologia , Filtração/métodos , Nanotecnologia/métodos , Solventes/farmacologia , Células Vero
18.
J Biosci Bioeng ; 128(2): 218-225, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30904455

RESUMO

Decellularization of a whole organ is an attractive process that has been used to create 3D scaffolds structurally and micro-architecturally similar to the native one. Currently used decellularization protocols exhibit disrupted extracellular matrix (ECM) structure and denatured ECM proteins. Therefore, maintaining a balance between ECM preservation and cellular removal is a major challenge. The aim of this study was to optimize a multistep Triton X-100 based protocol (either using Triton X-100/ammonium hydroxide mixture alone or after its modification with DNase, sodium dodecyl sulfate or trypsin) that could achieve maximum decellularization with minimal liver ECM destruction suitable for subsequent organ implantation without immune rejection. Based on our findings, Triton X-100 multistep protocol was insufficient for whole liver decellularization and needed to be modified with other detergents. Among all Triton X-100 modified protocols, a Triton X-100/DNase-based one was considered the most suitable. It maintains a gradual but sufficient removal of cells to generate decellularized biocompatible liver scaffolds without any significant alteration to ECM micro- and ultra-structure.


Assuntos
Materiais Biocompatíveis , Fígado/citologia , Engenharia Tecidual/métodos , Animais , Detergentes/farmacologia , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Octoxinol/farmacologia , Dodecilsulfato de Sódio/farmacologia , Tripsina/metabolismo
19.
Biochem Biophys Res Commun ; 512(2): 314-318, 2019 04 30.
Artigo em Inglês | MEDLINE | ID: mdl-30890336

RESUMO

Plasmin is a potent serin protease involved in a variety of biological functions, such as fibrinolysis and tissue remodeling. On performing an in vitro control assay to measure the activity of endogenous plasmin in cell lysates, a stimulatory effect of non-ionic detergent NP-40 on plasmin activity was discovered. Another non-ionic detergent, TX-100, also enhanced plasmin activity, while ionic detergents sodium deoxycholate and sodiem dodecyl sulfate abolished plasmin enzyme activity. Kinetic analysis of plasmin activity in the presence of NP-40 and TX-100 demonstrated an increase in Vmax; however, there was no change in Km values, suggesting that these detergents stimulate plasmin activity in a non-competitive manner. Fibrin plate assay indicates that NP-40 and TX-100 functionally stimulate plasmin activity by showing a dose-dependent increase in fibrinolysis.


Assuntos
Detergentes/farmacologia , Fibrinolisina/efeitos dos fármacos , Fibrinolisina/metabolismo , Ácido Desoxicólico/farmacologia , Fibrinólise/efeitos dos fármacos , Humanos , Técnicas In Vitro , Cinética , Octoxinol/farmacologia , Dodecilsulfato de Sódio/farmacologia
20.
PLoS One ; 14(3): e0199484, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30856175

RESUMO

Sodium dodecyl sulfate is a detergent that disrupts cell membranes, activates cell wall integrity signaling and restricts cell growth in Saccharomyces cerevisiae. However, the underlying mechanism of how sodium dodecyl sulfate inhibits cell growth is not fully understood. Previously, we have shown that deletion of the MCK1 gene leads to sensitivity to sodium dodecyl sulfate; thus, we implemented a suppressor gene screening revealing that the overexpression of TAT2 tryptophan permease rescues cell growth in sodium dodecyl sulfate-treated Δmck1 cells. Therefore, we questioned the involvement of tryptophan in the response to sodium dodecyl sulfate treatment. In this work, we show that trp1-1 cells have a disadvantage in the response to sodium dodecyl sulfate compared to auxotrophy for adenine, histidine, leucine or uracil when cells are grown on rich media. While also critical in the response to tea tree oil, TRP1 does not avert growth inhibition due to other cell wall/membrane perturbations that activate cell wall integrity signaling such as Calcofluor White, Congo Red or heat stress. This implicates a distinction from the cell wall integrity pathway and suggests specificity to membrane stress as opposed to cell wall stress. We discovered that tyrosine biosynthesis is also essential upon sodium dodecyl sulfate perturbation whereas phenylalanine biosynthesis appears dispensable. Finally, we observe enhanced tryptophan import within minutes upon exposure to sodium dodecyl sulfate indicating that these cells are not starved for tryptophan. In summary, we conclude that internal concentration of tryptophan and tyrosine makes cells more resistant to detergent such as sodium dodecyl sulfate.


Assuntos
Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , Dodecilsulfato de Sódio/farmacologia , Triptofano/metabolismo , Sistemas de Transporte de Aminoácidos/genética , Sistemas de Transporte de Aminoácidos/metabolismo , Transporte Biológico Ativo/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Detergentes/farmacologia , Farmacorresistência Fúngica/genética , Farmacorresistência Fúngica/fisiologia , Genes Fúngicos , Quinase 3 da Glicogênio Sintase/genética , Quinase 3 da Glicogênio Sintase/metabolismo , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Estresse Fisiológico/efeitos dos fármacos
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