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1.
Sensors (Basel) ; 21(3)2021 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-33513861

RESUMO

In this paper, a novel optical technique for following the progress of the blister copper desulfurization process is presented. The technique is based on the changes observed in the continuous spectrum of the visible-near-infrared (VIS-NIR) radiation that the blister melt emits while the chemical reactions of the sulfur elimination process are taking place. Specifically, the proposed technique uses an optical probe composed of an optical fiber, a collimating lens, and a quartz tube, which is immersed in the melt. This optical probe provides a field of view of the blowing zone where the desulfurization reaction occurs. The experimental results show that the melt VIS-NIR total irradiance evolves inversely to the SO2 concentration reported by a gas analyzer based on differential optical absorption spectroscopy. Furthermore, the blister copper spectral emissivity as well as the total emissivity observed throughout the process show strong correlation with the sulfur content during desulfurization reaction.

2.
Pharmacogenomics J ; 20(4): 613-620, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32015454

RESUMO

Oseltamivir, a pro-drug, is the best option for treatment and chemoprophylaxis for influenza outbreaks. However, many patients treated with oseltamivir developed adverse reactions, including hypersensitivity, gastritis, and neurological symptoms. The aim of this study was to determine the adverse drug reactions (ADRs) in Mexican patients treated with oseltamivir and whether these ADRs are associated with SNPs of the genes involved in the metabolism, transport, and interactions of oseltamivir. This study recruited 310 Mexican patients with acute respiratory diseases and treated them with oseltamivir (75 mg/day for 5 days) because they were suspected to have influenza A/H1N1 virus infection. Clinical data were obtained from medical records and interviews. Genotyping was performed using real-time polymerase chain reaction and TaqMan probes. The association was assessed under genetic models with contingency tables and logistic regression analysis. Out of 310 patients, only 38 (12.25%) presented ADRs to oseltamivir: hypersensitivity (1.9%), gastritis (10%), and depression and anxiety (0.9%). The polymorphism ABCB1-rs1045642 was associated with adverse drug reactions under the recessive model (P = 0.017); allele C was associated with no adverse drug reactions, while allele T was associated with adverse drug reactions. The polymorphisms SLC15A1-rs2297322, ABCB1-rs2032582, and CES1-rs2307243 were not consistent with Hardy-Weinberg equilibrium, and no other associations were found for the remaining polymorphisms. In conclusion, the polymorphism rs1045642 in the transporter encoded by the ABCB1 gene is a potential predictive biomarker of ADRs in oseltamivir treatment.


Assuntos
Antivirais/metabolismo , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/genética , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/metabolismo , Oseltamivir/metabolismo , Polimorfismo de Nucleotídeo Único/genética , Transtornos Respiratórios/genética , Transtornos Respiratórios/metabolismo , Doença Aguda , Adolescente , Adulto , Antivirais/efeitos adversos , Transporte Biológico/fisiologia , Criança , Interações Medicamentosas/fisiologia , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/epidemiologia , Feminino , Estudos de Associação Genética/métodos , Humanos , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Influenza Humana/tratamento farmacológico , Influenza Humana/epidemiologia , Influenza Humana/genética , Influenza Humana/metabolismo , Masculino , México/epidemiologia , Pessoa de Meia-Idade , Oseltamivir/efeitos adversos , Transporte Proteico/fisiologia , Transtornos Respiratórios/tratamento farmacológico , Transtornos Respiratórios/epidemiologia , Estudos Retrospectivos , Adulto Jovem
4.
Sensors (Basel) ; 20(5)2020 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-32120881

RESUMO

In this paper, we report on the spectral detection of wustite, Fe(II) oxide (FeO), and magnetite, Fe(II, III) oxide (Fe3O4), molecular emissions during the combustion of pyrite (FeS2), in a laboratory-scale furnace operating at high temperatures. These species are typically generated by reactions occurring during the combustion (oxidation) of this iron sulfide mineral. Two detection schemes are addressed: the first consisting of measurements with a built-in developed spectrometer with a high sensitivity and a high spectral resolution. The second one consisting of spectra measured with a low spectral resolution and a low sensitivity commercial spectrometer, but enhanced and analyzed with post signal processing and multivariate data analysis such as principal component analysis (PCA) and a multivariate curve resolution - the alternating least squares method (MCR-ALS). A non-linear model is also proposed to reconstruct spectral signals measured during pyrite combustion. Different combustion conditions were studied to evaluate the capacity of the detection schemes to follow the spectral emissions of iron oxides. The results show a direct correlation between FeO and Fe3O4 spectral features intensity, and non-linear relations with key combustion variables such as flame temperature, and the combusted sulfide mineral particle size.

5.
Sensors (Basel) ; 18(7)2018 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-29932156

RESUMO

In this paper, we report on spectral features emitted by a reaction shaft occurring in flash smelting of copper concentrates containing sulfide copper minerals such as chalcopyrite (CuFeS2), bornite (Cu5FeS4) and pyrite (FeS2). Different combustion conditions are addressed, such as sulfur-copper ratio and oxygen excess. Temperature and spectral emissivity features are estimated for each case by using the two wavelength method and radiometric models. The most relevant results have shown an increasing intensity behavior for higher sulfur-copper ratios and oxygen contents, where emissivity is almost constant along the visible spectrum range for all cases, which validates the gray body assumption. CuO and FeO emission line features along the visible spectrum appear to be a sensing alternative for describing the combustion reactions.

6.
Curr Genet ; 61(3): 325-34, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25381155

RESUMO

This paper examines the impact that single and interacting environmental stress factors have on tolerance mechanisms, molecular ecology and the relationship with secondary metabolite production by a group of mycotoxigenic species of economic importance. Growth of these fungi (Aspergillus flavus, A.ochraceus, A.carbonarius, Penicillium nordicum and P. verrucosum) is influenced by water and temperature interactions and type of solute used to induce water stress. Such abiotic stresses are overcome by the synthesis of increased amounts of low molecular weight sugar alcohols, especially glycerol and erythritol, to enable them to remain active under abiotic stress. This is accompanied by increased expression of sugar transporter genes, e.g., in A. flavus, which provides the nutritional means of tolerating such stress. The optimum conditions of water activity (a w) × temperature stress for growth are often different from those for secondary metabolite production. The genes for toxin production are clustered together and their relative expression is influenced by abiotic interacting stress factors. For example., A. flavus synthesises aflatoxins under water stress in non-ionic solutes. In contrast, P. nordicum specifically occupies a high salt (0.87 a w = 22% NaCl) niche such as cured meats, and produces ochratoxin A (OTA). There is differential and temporal expression of the genes in the secondary metabolite clusters in response to a w × temperature stress. We have used a microarray and integrated data on growth, relative expression of key genes in the biosynthetic pathways for secondary metabolite production and toxin production using a mixed growth model. This was used to correlate these factors and predict the toxin levels produced under different abiotic stress conditions. This system approach to integrate these different data sets and model the relationships could be a powerful tool for predicting the relative toxin production under extreme stress conditions, including climate change scenarios. This approach will facilitate a better functional understanding of the influence that environmental stress has on these mycotoxigenic fungi and enable better prevention strategies to be developed based on this system-based approach.


Assuntos
Meio Ambiente , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Fungos/fisiologia , Metaboloma , Família Multigênica , Metabolismo Secundário/genética , Estresse Fisiológico , Regulação Fúngica da Expressão Gênica , Interação Gene-Ambiente , Modelos Biológicos , Fenótipo , Temperatura , Água
7.
J Forensic Sci ; 69(4): 1171-1182, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38798041

RESUMO

Skeletal evidence usually constitutes the only source of information to interpret lesion patterns that help to clarify the circumstances surrounding death. The examination and interpretation of bone trauma are essential to the application and utility of anthropology as a forensic science. When discussing the effect of gunshot wounds in bone, it becomes imperative to differentiate between short and long-distance injuries based on clear, distinct, and observable signs. To contribute to the debate, our focus is directed toward the external analysis of the so-called circumferential delamination defect (CDD) as an observable proxy for close-range shooting (≤30 cm) and contact gunshot wounds in the skull. In the context of known extrajudicial killings, in which the perpetrators used short 9 × 19 FMJ ammunition in a close-range shooting, instances of CDD have been documented. Empirical evidence reinforcing the causal relationship between CDD and close-range shootings is presented. Elements' characteristics of firearm residues were also found in remains buried for up to 30 years. Primarily, this work shows that the concentrations of gunshot residues (Pb, Ba, and Sb) resemble those observed in fresh corpses with the same gunshot wound (GSW). Moreover, the correlation observed between CDD and gunshot residues, where the likelihood of CDD increases the closer to the head and the more perpendicular the shot angle is, reinforces CDD as a pivotal discriminatory factor in the skeletal evidence of short-range or contact shot. This research contributes to the field of forensic anthropology by providing fundamental insights into the etiology of CDD and its practical application.


Assuntos
Armas de Fogo , Balística Forense , Traumatismos Cranianos Penetrantes , Ferimentos por Arma de Fogo , Humanos , Ferimentos por Arma de Fogo/patologia , Masculino , Traumatismos Cranianos Penetrantes/patologia , Bário/análise , Chumbo/análise , Adulto , Pessoa de Meia-Idade , Homicídio , Crânio/lesões , Crânio/patologia , Cicloexanonas
8.
Dent Mater J ; 2024 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-38960669

RESUMO

Orthodontic brackets and archwires placed intraorally are subject to corrosion, leading to the release of cytotoxic metal ions. The aim of this study was to determine whether the use of orthodontic NiTi archwires increases systemic Ni levels and cause alterations on the DNA of cells unrelated to the oral environment such as lymphocytes and sperm cells. Human urine, semen and blood samples were collected before (baseline) sham placement of orthodontic archwires and 15 and 30 days after placement. Lymphocytes and sperm cells cells were evaluated by comet assay. Ni concentration levels in urine increased significantly between baseline and 15 days (p<0.01) and 15 and 30 days of exposure (p<0.01). Progressive decrease in sperm viability and motility was observed between the sampling periods. Lymphocytes and sperm cells showed DNA fragmentation. The increase in systemic concentration of nickel induced structural damage in the DNA of lymphocytes and human sperm cells.

9.
Forensic Sci Int ; 343: 111564, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36669240

RESUMO

Since the publication of Lamendin's age estimation technique, the root dentin translucency has received increasing attention as an important indicator of age. Recently, Parra and colleagues presented the Forensic International Dental Database (FIDB), a proposal to estimate age at death in adults based on Bayes theorem by applying the criteria of Lamendin's technique. The present study aims to update the procedure and to evaluate a new version of the method (named FIDBv2) using two control samples from Colombia and Greece. The performance of this new version was acceptable and suggests that the method is suitable for age at death estimation in adult individuals from different forensic contexts. The best approximations to chronological age were obtained for individuals between 30 and 60 years old, with errors less than 10 years. The age estimations calculated on control samples suggest the adequate performance of FIDBv2 on individuals from varied populations. It can be stated that the FIDBv2 constitutes a solid alternative to be used in contexts where no additional data are available. Here we reinforce the initial idea that this model for estimating age at death in adults may be generalizable to any forensic context in the world.


Assuntos
Determinação da Idade pelos Dentes , Adulto , Humanos , Pessoa de Meia-Idade , Teorema de Bayes , Determinação da Idade pelos Dentes/métodos , Raiz Dentária , Medicina Legal , Dentina , Odontologia Legal/métodos
10.
J Forensic Sci ; 66(4): 1520-1523, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33645633

RESUMO

International protocols for forensic investigations are often created by committee and in isolation. When field tested, the results of such tests are rarely reported to the wider forensic community. This study presents a comparative study of one such protocol, the International Protocol on the Documentation and Investigation of Sexual Violence in Conflict: Best Practice in the Documentation of Sexual Violence as a Crime or Violation of International Law. The protocol was used in a pilot study involving 20 victims of conflict-related sexual assault in the Democratic Republic of the Congo (DRC). The results of the pilot study were compared with an existing database of 341 victims of sexual assault (also from the same region of the DRC) who were examined using another protocol developed and utilized by Medicins Sans Frontier (MSF). The results clearly indicate the international protocol was far superior in all aspects, including comprehensive data capture and ease-of-use. Although the MSF protocol is intended for humanitarian purposes, all medical records are subject to potential downstream forensic applications. Given constraints in funding and resources in conflict zones, the wide-spread adoption of the full international protocol would ensure that every victim receives a complete, forensically valid examination suitable for the future pursuit of justice.


Assuntos
Vítimas de Crime , Medicina Legal/métodos , Delitos Sexuais , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Conflitos Armados , Criança , Estudos de Viabilidade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Exame Físico , Projetos Piloto , Adulto Jovem
11.
Anal Chim Acta ; 1178: 338805, 2021 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-34482864

RESUMO

The development of real-time monitoring sensors for pyro-metallurgical processes is an analytical challenge, mainly due to adverse environmental conditions, high spectral interferences and multiphase (molten and gas) reactions. This work demonstrates the suitability of stand-off LIBS (ST-LIBS) for real time monitoring of the desulfurization of blister copper which is carried out in molten phase. Here sulfur is removed by the formation of SO2 by supplying oxygen in molten phase. Using ST-LIBS the relative emission intensities of Cu(I) at 351.06 nm, O at 777.34 nm and S at 921.29 nm in both molten and gaseous phase were considered simultaneously during the process. This was possible only by the use high energy laser pulse over up to 270 mJ per pulse. In the case of copper, the selection of emission lines was assessed considering non-linear behavior, which is caused by self-absorption. For the first time, real time determination of sulfur in ppm range is reported by ST-LIBS using low sensitive lines from the NIR region. These results were validated with differential optical absorption spectroscopy (DOAS) as gold standard method. The analytical information obtained by LIBS can precisely determine the critical end-point of the desulfurization where the removal of sulfur is finished, and copper started to oxidize.


Assuntos
Vesícula , Cobre , Humanos , Lasers , Análise Espectral , Enxofre
12.
J Forensic Sci ; 66(6): 2456-2468, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34723398

RESUMO

In forensic science, the information that teeth provide to the age estimation process is very important. In adults, one of the most widely used indicators of skeletal age is the Root Dentin Translucency (RDT), mainly through the Lamendin technique, which is used in various Latin American contexts. Recently, Parra et al. (2020) have developed a Bayesian regression model using the Lamendin technique to establish standardized criteria for estimating age-at-death in adults in various forensic contexts. In this study, we evaluate the applicability of this proposal together with the proposal by Lamendin et al. (1992) and Prince and Ubelaker (2002) in Latin American contexts. A sample of single-rooted teeth belonging to 805 individuals from six Latin American countries was used. The results of the three proposals considered were analyzed taking into account factors such as age, sex, origin, and the tooth surface on which the variables were surveyed. Of the factors that would affect the estimates, it was found that the age of the individuals had the greatest influence. However, it was confirmed that the sex and surface of the teeth on which the measurements were taken did not influence the final result. On the other hand, as we expected, the application of the analyzed proposals would also be possible in other forensic contexts, as shown by the results obtained according to the origin. This research expands the FIDB with more information on Latino contexts.


Assuntos
Determinação da Idade pelos Dentes/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Bases de Dados Factuais , Feminino , Odontologia Legal/métodos , Retração Gengival/patologia , Humanos , América Latina , Luz , Masculino , Pessoa de Meia-Idade , Raiz Dentária/anatomia & histologia , Adulto Jovem
13.
Forensic Sci Int ; 317: 110499, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32947240

RESUMO

The impetus to begin a legal investigation or prosecution of the crime of genocide is a "trigger mechanism," which serves as the prima facie case against the accused state or actor. Unlike domestic cases of homicide, the trigger mechanisms for international genocide investigations to date have not included any forensic evidence nor have they sought input from forensic scientists. The jurists tasked with addressing the trigger mechanisms were fully capable of assessing forensic evidence but unable to generate it. This study reviews four recent large-scale investigations of genocide in Bosnia, Rwanda, Darfur and Myanmar to reveal pitfalls in the cases that could have been avoided by the earlier inclusion of scientific expertise, and identifies the potential contributions of forensic science in future investigations.


Assuntos
Ciências Forenses/legislação & jurisprudência , Genocídio/legislação & jurisprudência , Bósnia e Herzegóvina , Humanos , Direito Internacional , Mianmar , Ruanda , Sudão , Nações Unidas
14.
Forensic Sci Int ; 317: 110572, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33232857

RESUMO

Estimation of the age-at-death in adults is essential when the identification of deceased persons with unknown identity is required in both humanitarian and judicial contexts. However, the methodologies and the results obtained can be questioned. Various efforts have been developed to adjust procedures to specific populations, always seeking the precision and accuracy of the methodologies. It is known that the estimation of the age-at-death in adults coexists with wide margins of error, due to several reasons, including but not limited to statistical problems, the size of the sample or the physiological process of aging. This research focuses on a degenerative indicator of the dentin (Root Dentin Translucency) and its combination with Periodontal Height (PH) following the Lamendin's technique for estimation of the age-at-death in adults. The main objective of this research was to demonstrate the applicability of a Bayesian model based on a Forensic International Dental Database (FIDB) that include Root Translucency Height (RTH) and PH as a method to age-at-death in adults. The conclusion of this research was that the combined both indicators become a generalizable age-at-death in adults model for all human populations, where the Bayesian method would offer optimal results in any population. In this way, those populations that do not have had the possibility of validating a specific procedure, now have the opportunity to apply a valid method for estimating age-at-death in adults to global scope.


Assuntos
Determinação da Idade pelos Dentes/métodos , Cor , Dentina/anatomia & histologia , Raiz Dentária/anatomia & histologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Bases de Dados Factuais , Feminino , Odontologia Legal/métodos , Humanos , Funções Verossimilhança , Masculino , Pessoa de Meia-Idade , Adulto Jovem
15.
J Forensic Sci ; 53(3): 608-11, 2008 May.
Artigo em Inglês | MEDLINE | ID: mdl-18471205

RESUMO

Methods of estimating adult age at death from such dental features as root translucency, root length, and the extent of periodontosis in intact single rooted teeth have proven useful in studies of modern individuals, especially in estimating age in older adults. Questions remain however, concerning the impact of regional and population variation on accuracy. To examine this issue, three approaches to age estimation were applied to a diverse sample of 100 individuals from Peru. Mean errors of estimation for all three approaches were similar to those originally reported, suggesting minimal impact of population variation of the features measured. A new regression equation, specifically for Peruvian samples, is provided.


Assuntos
Determinação da Idade pelos Dentes/métodos , Odontologia Legal/métodos , Raiz Dentária/anatomia & histologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Modelos Estatísticos , Peru
16.
Int J Biol Macromol ; 107(Pt B): 1629-1640, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29055703

RESUMO

Laccases are enzymes that have the ability to catalyze the oxidation of a wide spectrum of phenolic compounds with the four-electron reduction of molecular oxygen to water. The active site of those proteins contains four copper ions, classified into three types. Laccases are interesting enzymes for study from the point of view of their structure, function and application because of their role in lignin degradation. Structural studies of two thermostable laccases produced by the strain Pycnoporus sanguineus CS43 (PsLacI and PsLacII) were performed. Both isoforms of PsLac show high thermal stability, at 60°C and 50°C, respectively, and they remained active at a high concentration of organic solvents. However, PsLacI has a higher thermal and pH stability and tolerance against inhibitors, and is a more efficient catalyst for ABTS and DMP (laccases substrate) than PsLacII. Based on the determined crystal structures we achieved insights into the structural factors relevant for the enzymatic properties of PsLacI and PsLacII. N-glycosylation site Asn354, which is very often present in structures of fungal laccases from other species, was not present in PsLac. This observation may be of particular significance due to the close distance between Asn354 and the substrate-binding pocket. This results in better access to the hydrophobic cavity for a particular substrate. Furthermore, we identified significant differences in the region of substrate-binding pocket, which confer PsLacI a markedly better performance than PsLacII.


Assuntos
Lacase/química , Lacase/metabolismo , Pycnoporus/enzimologia , Temperatura , Sequência de Aminoácidos , Antraquinonas/isolamento & purificação , Carboidratos/química , Domínio Catalítico , Estabilidade Enzimática , Glicosilação , Isoenzimas/metabolismo , Modelos Moleculares , Multimerização Proteica , Estrutura Secundária de Proteína , Eletricidade Estática , Homologia Estrutural de Proteína , Especificidade por Substrato , Difração de Raios X
17.
Protein Pept Lett ; 24(2): 118-125, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-27491380

RESUMO

Marine species are comprising about a half of the whole global biodiversity; the sea offers an enormous resource for novel bioactive compounds. Several of the marine origin species show multifunctional bioactivities and characteristics that are useful for a discovery and/or reinvention of biologically active compounds. For millennia, marine species that includes cyanobacteria (blue-green algae) and red algae have been targeted to explore their enormous potential candidature status along with a wider spectrum of novel applications in bio- and non-bio sectors of the modern world. Among them, cyanobacteria are photosynthetic prokaryotes, phylogenetically a primitive group of Gramnegative prokaryotes, ranging from Arctic to Antarctic regions, capable of carrying out photosynthesis and nitrogen fixation. In the recent decade, a great deal of research attention has been paid on the pronouncement of bio-functional proteins along with novel peptides, vitamins, fine chemicals, renewable fuel and bioactive compounds, e.g., phycobiliproteins from marine species, cyanobacteria and red algae. Interestingly, they are extensively commercialized for natural colorants in food and cosmetics, antimicrobial, antioxidant, anti-inflammatory, neuroprotective, hepatoprotective agents and fluorescent neo-glycoproteins as probes for single particle fluorescence imaging fluorescent applications in clinical and immunological analysis. However, a comprehensive knowledge and technological base for augmenting their commercial utilities are lacking. Therefore, this paper will provide an overview of the phycobiliproteins-based research literature from marine cyanobacteria and red algae. This review is also focused towards analyzing global and commercial activities with application oriented-based research. Towards the end, the information is also given on the potential biotechnological and biomedical applications of phycobiliproteins.


Assuntos
Cianobactérias/metabolismo , Ficobiliproteínas/metabolismo , Rodófitas/metabolismo , Organismos Aquáticos/metabolismo , Indústria Farmacêutica , Indústria Alimentícia , Humanos , Imagem Óptica , Ficobiliproteínas/farmacologia
18.
J Microbiol Methods ; 128: 74-79, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27432343

RESUMO

The use of Nile Red for rapid monitoring of the neutral lipid content in microalgae has gained interest over the last decade, since neutral lipids are feedstock for renewable transportation fuel. In this review, we discuss the main considerations needed to make an NR protocol reliable for staining neutral lipids in microalgae. Cell wall permeability must be enhanced by using stain carriers: DMSO (5% v/v to 25% v/v), glycerol (0.1 to 0.125mg/mL), or EDTA (3.0 to 3.8mg/mL). Temperatures between 30 and 40°C facilitate the diffusion of NR through the cell wall without incurring excess quenching. Good NR-lipid interaction requires using a low NR/cell ratio; the NR concentration must be between 0.25µg/mL and 2.0µg/mL, and the cell concentration >5×10(4)cells/mL. In order to have the maximum and stable NR fluorescence, it is necessary to scan the excitation/emission wavelengths for up to a 40-min of incubation time. We outline a five-step method to customize the Nile Red protocol to a specific strain: 1) Evaluate the strain's suitability by checking for the presence of neutral lipid, 2) Select of the best excitation/emission wavelength, 3) Optimization of incubation time, stain carrier, dye concentration, and temperature, 4) Prepare single-strain algal cultures with different lipid contents to calibrate NR fluorescence with neutral-lipid content, and 5) Correlate NR fluorescence intensity to neutral lipid content for the same strain. Once the protocol is customized, the NR method allows for rapid and reliable monitoring of neutral lipid content of a microalgae strain.


Assuntos
Corantes Fluorescentes/química , Lipídeos/análise , Microalgas/química , Oxazinas/química , Calibragem , Parede Celular/química , Dimetil Sulfóxido/química , Ácido Edético/química , Glicerol/química , Temperatura
19.
J Zhejiang Univ Sci B ; 15(4): 343-52, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24711355

RESUMO

The production of thermostable laccases from a native strain of the white-rot fungus Pycnoporus sanguineus isolated in Mexico was enhanced by testing different media and a combination of inducers including copper sulfate (CuSO4). The best conditions obtained from screening experiments in shaken flasks using tomato juice, CuSO4, and soybean oil were integrated in an experimental design. Enhanced levels of tomato juice as the medium, CuSO4 and soybean oil as inducers (36.8% (v/v), 3 mmol/L, and 1% (v/v), respectively) were determined for 10 L stirred tank bioreactor runs. This combination resulted in laccase titer of 143,000 IU/L (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), pH 3.0), which represents the highest activity so far reported for P. sanguineus in a 10-L fermentor. Other interesting media resulting from the screening included glucose-bactopeptone which increased laccase activity up to 20,000 IU/L, whereas the inducers Acid Blue 62 and Reactive Blue 19 enhanced enzyme production in this medium 10 times. Based on a partial characterization, the laccases of this strain are especially promising in terms of thermostability (half-life of 6.1 h at 60 °C) and activity titers.


Assuntos
Lacase/biossíntese , Pycnoporus/enzimologia , Reatores Biológicos , Biotecnologia , Meios de Cultura , Indução Enzimática , Estabilidade Enzimática , Temperatura Alta , Concentração de Íons de Hidrogênio , Microbiologia Industrial , México , Pycnoporus/isolamento & purificação
20.
J R Soc Interface ; 10(85): 20130320, 2013 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-23697716

RESUMO

The objective of this study was to integrate data on the effect of water activity (a(w); 0.995-0.93) and temperature (20-35 °C) on activation of the biosynthetic FUM genes, growth and the mycotoxins fumonisin (FB1, FB2) by Fusarium verticillioides in vitro. The relative expression of nine biosynthetic cluster genes (FUM1, FUM7, FUM10, FUM11, FUM12, FUM13, FUM14, FUM16 and FUM19) in relation to the environmental factors was determined using a microarray analysis. The expression was related to growth and phenotypic FB1 and FB2 production. These data were used to develop a mixed-growth-associated product formation model and link this to a linear combination of the expression data for the nine genes. The model was then validated by examining datasets outside the model fitting conditions used (35 °C). The relationship between the key gene (FUM1) and other genes in the cluster (FUM11, FUM13, FUM9, FUM14) were examined in relation to aw, temperature, FB1 and FB2 production by developing ternary diagrams of relative expression. This model is important in developing an integrated systems approach to develop prevention strategies to control fumonisin biosynthesis in staple food commodities and could also be used to predict the potential impact that climate change factors may have on toxin production.


Assuntos
Fumonisinas/metabolismo , Fusarium/fisiologia , Regulação Fúngica da Expressão Gênica/fisiologia , Interação Gene-Ambiente , Temperatura Alta , Modelos Biológicos , Micotoxinas/biossíntese , Genes Fúngicos/fisiologia , Família Multigênica/fisiologia , Micotoxinas/genética
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