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1.
Braz. j. biol ; 83: e243874, 2023. graf
Article in English | MEDLINE, LILACS, VETINDEX | ID: biblio-1285606

ABSTRACT

Abstract In recent days, cheapest alternative carbon source for fermentation purpose is desirable to minimize production cost. Xylanases have become attractive enzymes as their potential in bio-bleaching of pulp and paper industry. The objective of the present study was to identify the potential ability on the xylanase production by locally isolated Bacillus pumilus BS131 by using waste fiber sludge and wheat bran media under submerged fermentation. Culture growth conditions were optimized to obtain significant amount of xylanase. Maximum xylanase production was recorded after 72 hours of incubation at 30 °C and 7 pH with 4.0% substrate concentration. In the nutshell, the production of xylanase using inexpensive waste fiber sludge and wheat-bran as an alternative in place of expensive xylan substrate was more cost effective and environment friendly.


Resumo Nos últimos dias, a fonte alternativa de carbono mais barata para fins de fermentação é desejável para minimizar o custo de produção. As xilanases têm se tornado enzimas atraentes como seu potencial no biobranqueamento da indústria de papel e celulose. O objetivo do presente estudo foi identificar a capacidade potencial na produção de xilanase por Bacillus pumilus BS131 isolado localmente usando lodo de fibra residual e farelo de trigo em meio de fermentação submersa. As condições de crescimento da cultura foram otimizadas para obter uma quantidade significativa de xilanase. A produção máxima de xilanase foi registrada após 72 horas de incubação a 30 °C e pH 7 com concentração de substrato de 4,0%. Resumindo, a produção de xilanase usando lodo de fibra residual de baixo custo e farelo de trigo como uma alternativa no lugar do substrato de xilano caro foi mais econômica e ecológica.


Subject(s)
Bacillus/metabolism , Bacillus pumilus/metabolism , Sewage , Temperature , Dietary Fiber , Endo-1,4-beta Xylanases/metabolism , Fermentation , Hydrogen-Ion Concentration
2.
Braz. j. biol ; 82: e244735, 2022. tab, graf
Article in English | LILACS | ID: biblio-1249280

ABSTRACT

Abstract L-Asparaginase catalysing the breakdown of L-Asparagine to L-Aspartate and ammonia is an enzyme of therapeutic importance in the treatment of cancer, especially the lymphomas and leukaemia. The present study describes the recombinant production, properties and anticancer potential of enzyme from a hyperthermophilic archaeon Pyrococcus abyssi. There are two genes coding for asparaginase in the genome of this organism. A 918 bp gene encoding 305 amino acids was PCR amplified and cloned in BL21 (DE3) strain of E. coli using pET28a (+) plasmid. The production of recombinant enzyme was induced under 0.5mM IPTG, purified by selective heat denaturation and ion exchange chromatography. Purified enzyme was analyzed for kinetics, in silico structure and anticancer properties. The recombinant enzyme has shown a molecular weight of 33 kDa, specific activity of 1175 U/mg, KM value 2.05mM, optimum temperature and pH 80°C and 8 respectively. No detectable enzyme activity found when L-Glutamine was used as the substrate. In silico studies have shown that the enzyme exists as a homodimer having Arg11, Ala87, Thr110, His112, Gln142, Leu172, and Lys232 being the putative active site residues. The free energy change calculated by molecular docking studies of enzyme and substrate was found as ∆G - 4.5 kJ/mole indicating the affinity of enzyme with the substrate. IC50 values of 5U/mL to 7.5U/mL were determined for FB, caco2 cells and HepG2 cells. A calculated amount of enzyme (5U/mL) exhibited 78% to 55% growth inhibition of caco2 and HepG2 cells. In conclusion, the recombinant enzyme produced and characterized in the present study offers a good candidate for the treatment of cancer. The procedures adopted in the present study can be prolonged for in vivo studies.


Resumo A L-asparaginase, que catalisa a degradação da L-asparagina em L-aspartato e amônia, é uma enzima de importância terapêutica no tratamento do câncer, especialmente dos linfomas e da leucemia. O presente estudo descreve a produção recombinante, propriedades e potencial anticancerígeno da enzima de Pyrococcus abyssi, um archaeon hipertermofílico. Existem dois genes que codificam para a asparaginase no genoma desse organismo. Um gene de 918 bp, que codifica 305 aminoácidos, foi amplificado por PCR e clonado na cepa BL21 (DE3) de E. coli usando o plasmídeo pET28a (+). A produção da enzima recombinante foi induzida sob 0,5mM de IPTG, purificada por desnaturação seletiva por calor e cromatografia de troca iônica. A enzima purificada foi analisada quanto à cinética, estrutura in silico e propriedades anticancerígenas. A enzima recombinante apresentou peso molecular de 33 kDa, atividade específica de 1.175 U / mg, valor de KM 2,05 mM, temperatura ótima de 80º C e pH 8. Nenhuma atividade enzimática detectável foi encontrada quando a L-glutamina foi usada como substrato. Estudos in silico mostraram que a enzima existe como um homodímero, com Arg11, Ala87, Thr110, His112, Gln142, Leu172 e Lys232 sendo os resíduos do local ativo putativo. A mudança de energia livre calculada por estudos de docking molecular da enzima e do substrato foi encontrada como ∆G - 4,5 kJ / mol, indicando a afinidade da enzima com o substrato. Valores de IC50 de 5U / mL a 7,5U / mL foram determinados para células FB, células caco2 e células HepG2. Uma quantidade de enzima (5U / mL) apresentou inibição de crescimento de 78% a 55% das células caco2 e HepG2, respectivamente. Em conclusão, a enzima recombinante produzida e caracterizada no presente estudo é uma boa possibilidade para o tratamento do câncer. Os procedimentos adotados na presente pesquisa podem ser aplicados para estudos in vivo.


Subject(s)
Humans , Asparaginase/biosynthesis , Asparaginase/pharmacology , Pyrococcus abyssi/enzymology , Antineoplastic Agents/pharmacology , Substrate Specificity , Enzyme Stability , Recombinant Proteins/biosynthesis , Recombinant Proteins/pharmacology , Caco-2 Cells , Escherichia coli/genetics , Molecular Docking Simulation , Hydrogen-Ion Concentration
3.
Braz. j. biol ; 82: e239449, 2022. tab, graf
Article in English | LILACS | ID: biblio-1249271

ABSTRACT

Abstract Alpha amylase, catalyzing the hydrolysis of starch is a ubiquitous enzyme with tremendous industrial applications. A 1698 bp gene coding for 565 amino acid amylase was PCR amplified from Geobacillus thermodenitrificans DSM-465, cloned in pET21a (+) plasmid, expressed in BL21 (DE3) strain of E. coli and characterized. The recombinant enzyme exhibited molecular weight of 63 kDa, optimum pH 8, optimum temperature 70°C, and KM value of 157.7µM. On pilot scale, the purified enzyme efficiently removed up to 95% starch from the cotton fabric indicating its desizing ability at high temperature. 3D model of enzyme built by Raptor-X and validated by Ramachandran plot appeared as a monomer having 31% α-helices, 15% β-sheets, and 52% loops. Docking studies have shown the best binding affinity of enzyme with amylopectin (∆G -10.59). According to our results, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276, and Arg175 constitute the potential active site of enzyme.


Resumo A alfa-amilase, que catalisa a hidrólise do amido, é uma enzima ubíqua com imensas aplicações industriais. Um gene de 1698 pb que codifica a amilase de 565 aminoácidos foi amplificado por PCR, a partir de Geobacillus thermodenitrificans DSM-465, clonado no plasmídeo pET21a (+), expresso na cepa BL21 (DE3) de E. coli e caracterizado. A enzima recombinante exibiu peso molecular de 63 kDa, pH ótimo igual a 8, temperatura ótima de 70° C e valor KM de 157,7 µM. Em escala piloto, a enzima purificada removeu com eficiência até 95% de amido do tecido de algodão, indicando sua capacidade de desengomagem em alta temperatura. O modelo 3D da enzima construída por Raptor-X e validada por Ramachandran plot apareceu como um monômero com 31% de hélices alfa, 15% de folhas beta e 52% de loops. Os estudos de docking mostraram melhor afinidade de ligação da enzima com amilopectina (∆G: - 10,59). De acordo com nossos resultados, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276 e Arg175 constituem o sítio ativo potencial da enzima.


Subject(s)
Escherichia coli/genetics , alpha-Amylases/genetics , alpha-Amylases/metabolism , Temperature , Enzyme Stability , Cloning, Molecular , Geobacillus , Hydrogen-Ion Concentration
4.
Braz. j. biol ; 82: e234413, 2022. tab, graf
Article in English | LILACS | ID: biblio-1249269

ABSTRACT

Abstract The viscera and other residues from fish processing are commonly discarded by the fishing industry. These by-products can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 U⋅mg-1. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.


Resumo As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 U⋅mg-1. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas


Subject(s)
Peptide Hydrolases , Stomach , Temperature , Hydrogen-Ion Concentration
5.
Braz. J. Pharm. Sci. (Online) ; 58: e18984, 2022. graf
Article in English | LILACS | ID: biblio-1364429

ABSTRACT

Interferon-ß-1a (INF-ß-1a) has gained significant attention due to its emerging applications in the treatment of different human diseases. Therefore, many researchers have attempted to produce it in large quantities and also in a biologically active form using different expression systems. In the present study, we aimed to improve the expression level of INF-ß-1a by Pichia pastoris using optimization of culture conditions. The codon-optimized INF-ß- 1a gene was cloned into pPICZαA plasmid under the control of alcohol oxidase I (AOX1) promoter. The protein expression was induced using different concentrations of methanol at different pHs and temperatures. The biological activity of produced protein was evaluated by anti-proliferative assay. The ideal culture conditions for the expression of INF-ß-1a by P. pastoris were found to be induction with 2% methanol at pH 7.0 culture medium at 30 C which yielded a concentration of 15.5 mg/L INF-ß-1a in a shake flask. Our results indicate that differences in glycosylation pattern could result in different biological activities as INF- ß-1a produced by P. pastoris could significantly more reduce the cell viability of HepG-2 cells, a hepatocellular carcinoma cell line, than a commercially available form of this protein produced by CHO


Subject(s)
Pichia/classification , Interferon-beta/agonists , Carcinoma, Hepatocellular/pathology , Process Optimization , Codon , Cells , Carcinoma, Hepatocellular , Hydrogen-Ion Concentration
6.
Braz. J. Pharm. Sci. (Online) ; 58: e18946, 2022. tab, graf
Article in English | LILACS | ID: biblio-1364411

ABSTRACT

Abstract To investigate structure-property relationship of polymer-based curcumin solid dispersion (SD), three acrylic polymers were used to formulate curcumin SD by solvent evaporation method. Curcumin Eudragit EPO SD (cur@EPO), curcumin Eudragit RS PO SD (cur@RSPO) and curcumin Eudragit RL PO SD (cur@RLPO) showed deep red, golden orange and reddish orange color, respectively. Cur@RSPO entrapped 15.42 wt% of curcumin followed by cur@RL PO and cur@EPO. FTIR spectra indicated that in cur@EPO, curcumin may transfer hydrogen to the dimethylaminoethyl methacrylate group and thus change its color to red. In contrast, curcumin may form hydrogen bonding with Eudragit RS PO and Eudragit RL. Curcumin exists in amorphous state in three SDs as proved by differential scanning calorimetry and X-Ray diffraction measurement. In vitro digestion presented that lower pH value in simulated gastric fluid (SGF) stimulates the curcumin release from cur@EPO while permeability influences the release profile in other two SDs. When in simulated intestinal fluid (SIF), first order release model governs the release behaviors of all three SDs which showed sustained release pattern. Our results are helpful to elucidate how structure of polymer may impact on the major properties of curcumin contained SD and will be promising to broaden its therapeutic applications.


Subject(s)
Polymers , Curcumin/analysis , Methods , Solvents/administration & dosage , X-Ray Diffraction/instrumentation , In Vitro Techniques/methods , Calorimetry, Differential Scanning/methods , Evaporation/classification , Spectroscopy, Fourier Transform Infrared , Color , Citrus sinensis/classification , Hydrogen-Ion Concentration
7.
Braz. j. biol ; 82: e234500, 2022. tab, graf
Article in English | LILACS | ID: biblio-1153464

ABSTRACT

Abstract This work aimed to obtain aspartic proteases of industrial and biotechnological interest from the stomach of the crevalle jack fish (Caranx hippos). In order to do so, a crude extract (CE) of the stomach was obtained and subjected to a partial purification by salting-out, which resulted in the enzyme extract (EE) obtainment. EE proteases were characterized physicochemically and by means of zymogram. In addition, the effect of chemical agents on their activity was also assessed. By means of salting-out it was possible to obtain a purification of 1.6 times with a yield of 49.4%. Two acid proteases present in the EE were observed in zymogram. The optimum temperature and thermal stability for EE acidic proteases were 55 ºC and 45 °C, respectively. The optimum pH and pH stability found for these enzymes were pH 1.5 and 7.0, respectively. Total inhibition of EE acid proteolytic activity was observed in the presence of pepstatin A. dithiothreitol (DTT) and Ca2+ did not promote a significant effect on enzyme activity. In the presence of heavy metals, such as Al3+, Cd2+ and Hg2+, EE acidic proteases showed more than 70% of their enzymatic activity. The results show that it is possible to obtain, from the stomach of C. hippos, aspartic proteases with high proteolytic activity and characteristics that demonstrate potential for industrial and biotechnological applications.


Resumo Este trabalho objetivou obter proteases aspárticas de interesse industrial e biotecnológico a partir do estômago do peixe xaréu (Caranx hippos). Para isso, foi obtido um extrato bruto do estômago, o qual foi submetido a uma purificação parcial por salting-out onde se obteve o extrato enzimático (EE). As proteases do EE foram caracterizadas físico-quimicamente e através de zimograma. Além disso, o efeito de agentes químicos sobre sua atividade também foi avaliado. Através de salting-out foi possível obter uma purificação de 1,6 vezes com rendimento de 49,4%. Foram observadas duas proteases ácidas presentes no EE através de zimograma. A temperatura ótima e a estabilidade térmica para as proteases ácidas do EE foram de 55 ºC e 45 °C, respectivamente. O pH ótimo e a estabilidade ao pH encontrados para estas enzimas foram o pH 1,5 e 7,0, respectivamente. Observou-se a inibição total da atividade proteolítica ácida do EE na presença de pepstatina A. O ditiotreitol (DTT) e o Ca2+ não promoveram efeito significativo na atividade enzimática. Na presença de metais pesados, como Al3+, Cd2+ e Hg2+, o EE manteve mais de 70% de atividade enzimática do EE. Os resultados mostram que é possível obter, a partir do estômago de C. hippos, proteases aspárticas com alta atividade proteolítica e características que demonstram potencial para aplicações industriais e biotecnológicas.


Subject(s)
Animals , Peptide Hydrolases , Temperature , Hydrogen-Ion Concentration
8.
Braz. j. biol ; 82: e244496, 2022. tab, graf
Article in English | MEDLINE, LILACS, VETINDEX | ID: biblio-1278474

ABSTRACT

Abstract Enzymes immobilized onto substrates with excellent selectivity and activity show a high stability and can withstand extreme experimental conditions, and their performance has been shown to be retained after repeated uses. Applications of immobilized enzymes in various fields benefit from their unique characteristics. Common methods, including adsorption, encapsulation, covalent attachment and crosslinking, and other emerging approaches (e.g., MOFs) of enzyme immobilization have been developed mostly in recent years. In accordance with these immobilization methods, the present review elaborates the application of magnetic separable nanoparticles and functionalized SBA-15 and MCM-41 mesoporous materials used in the immobilization of enzymes.


Resumo Enzimas imobilizadas em substratos com excelente seletividade e atividade apresentam alta estabilidade e podem suportar condições experimentais extremas, e seu desempenho foi mantido após repetidos usos. As aplicações de enzimas imobilizadas em vários campos se beneficiam de suas características únicas. Métodos comuns, incluindo adsorção, encapsulamento, ligação covalente e reticulação, e outras abordagens emergentes (por exemplo, MOFs) de imobilização de enzima, foram desenvolvidos principalmente nos últimos anos. De acordo com esses métodos de imobilização, a presente revisão elabora a aplicação de nanopartículas magnéticas separáveis ​​e materiais mesoporosos funcionalizados SBA-15 e MCM-41 usados ​​na imobilização de enzimas.


Subject(s)
Enzymes, Immobilized/metabolism , Magnetite Nanoparticles , Enzyme Stability , Adsorption , Hydrogen-Ion Concentration
9.
Rev. cuba. anestesiol. reanim ; 20(3): e720, 2021.
Article in Spanish | LILACS, CUMED | ID: biblio-1351985

ABSTRACT

Introducción: Las alteraciones del equilibrio ácido-base pueden ser de carácter primario. En la mayoría de los casos dependen de la complicación de una enfermedad preexistente. La frecuencia de estos trastornos es elevada, especialmente, en enfermos hospitalizados en las unidades de atención al paciente grave. Su aparición conlleva implicaciones pronósticas significativas. Objetivo: Sistematizar sobre el estado actual del manejo del equilibrio ácido-base. Método: Se realizó una revisión bibliográfica en la que se utilizaron las herramientas del método científico. Se examinó toda la bibliografía disponible publicada en los últimos cinco años y así, elaborar una síntesis crítica, acorde al criterio y las competencias de los autores sobre la temática. Resultados: Se expone la importancia de la evaluación clínica, que unida a los niveles de PCO2, y de exceso o déficit de bases en una gasometría arterial, permiten identificar el trastorno ácido base existente. Igualmente, se destaca que el CO2 tiene una función clave en el control de la ventilación, así como las modificaciones que produce al flujo sanguíneo cerebral, el pH y el tono adrenérgico. Otro aspecto importante fue la reciente práctica clínica de la "hipercapnia permisiva" para reducir el metabolismo tisular y de esta manera, mejorar la función del surfactante e impedir la nitración de las proteínas. Conclusiones: El manejo de los desequilibrios ácido-base debe ser del dominio de todos los profesionales vinculados a la asistencia médica, pues el retraso de su diagnóstico puede empeorar la evolución y el pronóstico de los pacientes graves(AU)


Introduction: Acid-base balance alterations can be of a primary nature. In most cases, they depend on the complication of a pre-existing disease. The frequency of these disorders is high, especially in patients hospitalized in critical care units. Its appearance carries significant prognostic implications. Objective: To systematize the current state of acid-base balance management. Method: A bibliographic review was carried out, for which the tools of the scientific method were used. All the available bibliography, published in the last five years, was examined; thus, a critical synthesis was prepared, according to the criteria and competences of the authors regarding the subject. Results: The importance of the clinical evaluation is exposed, which, together with PCO2 levels as well as excess or deficit of bases in an arterial blood gas, allow to identify the existing acid-base disorder. Likewise, it is highlighted that CO2 has a key function in ventilation control, together with the modifications it produces on cerebral blood flow, pH and adrenergic tone. Another important aspect was the recent clinical practice of "permissive hypercapnia" to reduce tissue metabolism and thus improve surfactant function and prevent protein nitration. Conclusions: The management of acid-base imbalances should be mastered by all professionals associated to medical care, since any delay in its diagnosis can worsen the evolution and prognosis of seriously ill patients(AU)


Subject(s)
Humans , Male , Female , Acid-Base Equilibrium , Acid-Base Imbalance , Blood Gas Analysis/methods , Critical Care , Medical Care , Hydrogen-Ion Concentration
11.
Electron. j. biotechnol ; 52: 30-34, July. 2021. ilus, tab, graf
Article in English | LILACS | ID: biblio-1283487

ABSTRACT

BACKGROUND: This study aimed to develop an amplification method of urea detection based on pHsensitive liposomes. RESULTS: The urease covalently immobilized on the magnetic particles and the pH-sensitive liposomes encapsulating ferricyanide were added to the cyclic-voltammeter cell solution where urea was distributed. The conversion of urea into carbonic acid seemed to induce a pH decrease that caused a reduction in the electrostatic repulsion between the headgroups of weakly acidic 1,2-dipalmitoyl-sn-glycero3-succinate. The reduction induced the liposomes to release potassium ferricyanide that was encapsulated inside. The effects of urea concentration and pH value were investigated. A specific concentration (0.5 mg/mL) of the urea solution was set to observe the response. The activity of urease was reversible with respect to the pH change between 7 and 5. The sensitivity of this detection was almost identical to the comparable techniques such as an enzyme-linked immunosorbent assay and a field-effect transistor. CONCLUSIONS: In summary, the methodology developed in this study was feasible as a portable, rapid, and sensitive method.


Subject(s)
Urea/analysis , Liposomes/chemistry , Urease/chemistry , Enzyme-Linked Immunosorbent Assay , Enzymes, Immobilized , Hydrogen-Ion Concentration
12.
Electron. j. biotechnol ; 52: 85-92, July. 2021. graf, tab
Article in English | LILACS | ID: biblio-1283600

ABSTRACT

BACKGROUND: Nonribosomal peptide synthases (NRPS) can synthesize functionally diverse bioactive peptides by incorporating nonproteinogenic amino acids, offering a rich source of new drug leads. The bacterium Escherichia coli is a well-characterized production host and a promising candidate for the synthesis of nonribosomal peptides, but only limited bioprocess engineering has been reported for such molecules. We therefore developed a medium and optimized process parameters using the design of experiments (DoE) approach. RESULTS: We found that glycerol is not suitable as a carbon source for rhabdopeptide production, at least for the NRPS used for this study. Alternative carbon sources from the tricarboxylic acid cycle achieved much higher yields. DoE was used to optimize the pH and temperature in a stirred-tank reactor, revealing that optimal growth and optimal production required substantially different conditions. CONCLUSIONS: We developed a chemically defined adapted M9 medium matching the performance of complex medium (lysogeny broth) in terms of product concentration. The maximum yield in the reactor under optimized conditions was 126 mg L-1, representing a 31-fold increase compared to the first shaking-flask experiments with M9 medium and glycerol as the carbon source. Conditions that promoted cell growth tended to inhibit NRPS productivity. The challenge was therefore to find a compromise between these factors as the basis for further process development.


Subject(s)
Peptide Synthases/metabolism , Bioreactors/microbiology , Escherichia coli , Temperature , Biotechnology , Carbon/metabolism , Models, Statistical , Electrophoresis, Polyacrylamide Gel , Bioengineering , Hydrogen-Ion Concentration
13.
Acta amaz ; 51(2): 129-138, jun. 2021.
Article in English | LILACS | ID: biblio-1353415

ABSTRACT

O conceito de rio continuo prediz que mudanças graduais na geomorfologia e hidrologia de um rio podem causar mudanças longitudinais na estrutura de comunidades aquáticas. Nesse contexto, avaliamos como variáveis ambientais (condutividade elétrica, pH, transparência e velocidade da água), fatores espaciais (distância da cabeceira e Moran's eigen vector maps ­ MEMs) e presença de barragens afetam as assembleias de peixes de pequeno porte ao longo de um trecho de 105 km do alto Rio Branco, um tributário do Rio Madeira, bacia Amazônica, Brasil. Coletas com rede de arrasto foram realizadas em 15 pontos a jusante e montante de barragens durante a estação seca de 2019. Capturamos um total de 4.330 indivíduos, distribuídos em três ordens, nove famílias e 26 espécies (e um indivíduo híbrido). Condutividade elétrica e pH foram afetados significativamente pela presença de barragens e pela distância da cabeceira. A riqueza de espécies e a abundância não variaram em função das variáveis ambientais. Ao contrário da riqueza de espécies, que não variou ao longo do trecho amostrado, a abundância apresentou uma relação negativa com a distância da cabeceira. A composição de espécies variou significativamente em resposta ao pH, a fatores espaciais lineares e à presença de barragens. Nossos resultados sugerem que a mudanças na composição de espécies de peixes de pequeno porte no alto Rio Branco ocorre devido à variação em características da água, em função da limitação na dispersão inerente das espécies e em resposta à presença de barragens(AU)


Subject(s)
Hydrographic Basins , Amazonian Ecosystem , Gradient , Hydrogen-Ion Concentration
14.
Rev. cient. odontol ; 9(2): e058, abr.-jun. 2021. ilus, tab
Article in Spanish | LIPECS, LILACS, LIPECS | ID: biblio-1254597

ABSTRACT

Objetivo: Comparar el pH de cuatro marcas de geles aclaradores a base de peróxido de hidrógeno de altas concentraciones (30%-35%) Whiteness HP Maxx (HPM), Lase Peroxide (LP), Whiteness HP Automixx (HPA)y Dash (DA), a través del tiempo de aplicación clínica (inicio, 15', 30' y 45'). Materiales y métodos: El estudio fue experimental in vitro. Se evaluaron 40 muestras (dientes bovinos) divididos en 4 grupos, uno para cada marca comercial de gel. Se preparó cada gel de acuerdo con las instrucciones del fabricante y se colocó una cantidad necesaria en la superficie vestibular; posteriormente, se registró el pH del gel con un pHmetro digital al inicio, 15, 30 y 45 minutos. Los datos se analizaron con las pruebas Anova, Friedman y Wilcoxon. Resultados: Hubo una tendencia a la disminución del pH desde el tiempo inicial de aplicación hasta el tiempo final, con excepción del grupo de la marca DA, el cual mostró que los valores del pH fueron aumentando a través del tiempo. En la marca HPM, existe una diferencia significativa entre el tiempo inicial de aplicación del gel y el resto de tiempos. En la marca LP, a partir de los 15' de aplicación, sí hay diferencia significativa entre los tiempos. En cuanto a la marca HPA, existieron diferencias significativas entre el tiempo inicial de aplicación y los demás tiempos. Finalmente, con la marca DA se encontró únicamente una diferencia significativa entre el tiempo inicial de aplicación y el tiempo final. Conclusiones: El pH de los geles aclaradores disminuyó a través del tiempo de aplicación clínica en todas las marcas estudiadas, con excepción de la marca Dash 30%, la cual mostró un aumento. (AU)


Objective: The aim of this in vitro study was to evaluate the pH of four bleaching agents based on high concentration hydrogen peroxide (30-35%) Whiteness HP Maxx (HPM), Lase Peroxide (LP), Whiteness HP Automixx (HPA) and Dash (DA) in different clinical periods (baseline, 15', 30' and 45'). Materials and methods: 40 specimens (bovine teeth) were divided into 4 groups; one group for each bleaching agent. Each bleaching agent was prepared according to the manufacturer's instructions and was applied on the vestibular surface. The pH of the bleaching agent was measured with a digital pH meter at baseline, 15, 30 and 45 minutes. ANOVA, Friedman and Wilcoxon tests were applied. Results: The pH values showed a trend to decreasing from the initial time of application to the final time, except for the DA group, which showed increasing pH values over time. The HPM group showed significant differences between baseline and the remaining periods. The LP group LP showed significant difference between 15' and the other periods. The HPA group showed significant differences between baseline and the remaining periods. Finally, the DA group, showed a significant difference between baseline and 45'. Conclusions: The pH values of 3 of the bleaching agents decreased over time, with the exception of Dash which increased in the different time periods. (AU)


Subject(s)
Tooth Bleaching , Hydrogen-Ion Concentration , Hydrogen Peroxide , In Vitro Techniques , Epidemiology, Experimental
15.
Electron. j. biotechnol ; 51: 79-87, May. 2021. tab, ilus, graf
Article in English | LILACS | ID: biblio-1343441

ABSTRACT

BACKGROUND: At present, cellulases are the most important enzymes worldwide, and their demand has been increasing in the industrial sector owing to their notable hydrolysis capability. RESULTS: In the present study, contrary to conventional techniques, three physical parameters were statistically optimized for the production of cellulase by thermophilic fungi by using response surface methodology (RSM). Among all the tested thermophilic strains, the best cellulase producing fungus was identified as Talaromyces thermophilus ­ both morphologically and molecularly through 5.8S/ITS rDNA sequencing. The central composite design (CCD) was used to evaluate the interactive effect of the significant factors. The CCD was applied by considering incubation period, pH, and temperature as the model factors for the present investigation. A second-order quadratic model and response surface method revealed that the independent variables including pH 6, temperature 50 C, and incubation period 72 h significantly influenced the production of cellulases. The analysis of variance (ANOVA) indicated that the established model was significant (P 0.05) and showed the high adequacy of the model. The actual and predicted values of CMCase and FPase activity showed good agreement with each other and also confirmed the validity of the designed model. CONCLUSIONS: We believe the present findings to be the first report on cellulase production by exploiting Kans grass (Saccharum spontaneum) as a substrate through response surface methodology by using thermophilic fungus, Talaromyces thermophilus.


Subject(s)
Talaromyces/metabolism , Cellulases/biosynthesis , Analysis of Variance , Saccharum , Fermentation , Hot Temperature , Hydrogen-Ion Concentration
16.
Rev. cuba. pediatr ; 93(1): e1205, ene.-mar. 2021. tab
Article in Spanish | LILACS, CUMED | ID: biblio-1251747

ABSTRACT

Introducción: El ensayo de muestras matutinas de orina pudiera mejorar el estudio de la función tubular en niños y adolescentes. Objetivo: Describir las tubulopatías diagnosticadas en niños y adolescentes después del ensayo de muestras matutinas de orina. Métodos: Se completó un estudio retrospectivo y analítico en el Laboratorio de Estudio de la Función Renal, Servicio de Laboratorio Clínico, Hospital Pediátrico Docente "Juan Manuel Márquez", con 70 informes de la función tubular hechos en muestras matutinas de orina de 56 probandos (varones: 50,0 por ciento; edad promedio: 4,3 ± 5,5 años; edades < 12 meses: 41,1 por ciento) atendidos entre 2015-2019 (ambos inclusive) que contenían los valores del filtrado glomerular, la excreción urinaria absoluta y fraccional de las sustancias de interés, la brecha aniónica, la presión parcial de los gases, y la acidez titulable, el pH, la densidad y la osmolaridad de los fluidos pertinentes. Los resultados obtenidos se integraron dentro de las construcciones de caso de varias tubulopatías. Resultados: La función tubular estaba conservada en el 41,1 por ciento de los probandos. La inmadurez tubular explicó los hallazgos en otros dos niños. La hipercalciuria idiopática (16,0 por ciento), la diabetes insípida de causa nefrogénica (8,9 por ciento) y la insuficiencia renal aguda (5,3 por ciento) fueron los hallazgos más frecuentes. En 14 de los probandos se diagnosticaron 10 tubulopatías que recorrieron el raquitismo carencial, la hipofosfatasia, la enfermedad de Leigh, el síndrome de Bartter, la enfermedad de Dent y la acidosis tubular I, II y IV. Conclusiones: El estudio tubular en muestras matutinas de orina permite el diagnóstico de importantes tubulopatías en las edades pediátricas(AU)


Introduction: The morning urine sample assay may improve the study of tubular function in children and adolescents. Objective: Describe the tubulopathies diagnosed in children and adolescents after the trial of morning urine samples. Methods: A retrospective and analytical study was completed at the Renal Function´s Study Laboratory, in the Clinical Laboratory Service at "Juan Manuel Marquez" Teaching Pediatric Hospital, with 70 reports of tubular function made in morning urine samples of 56 testees (males: 50.0 percent; average age: 4.3 ± to 5.5 years; ages< 12 months: 41.1 percent) attended from 2015 to 2019 (both inclusive) containing glomerular filtration values, absolute and fractional urinary excretion of substances of interest, anionic gap, partial gas pressure, and titrable acidity, pH, density and osmolarity of relevant fluids. The results obtained were integrated into the case constructions of various tubulopathies. Results: Tubular function was preserved in 41.1 percent of the testees. Tubular immaturity explained the findings in two other children. Idiopathic hypercalciuria (16.0 percent), nephrogenic diabetes insipidus (8.9 percent) and acute renal failure (5.3 percent) were the most frequent findings. In 14 of the testees, 10 tubulopathies were diagnosed were through deficiency rickets, hypophosphatasia, Leigh's disease, Bartter syndrome, Dent disease and tubular acidosis I, II and IV.. Conclusions: The tubular study with morning urine samples allows the diagnosis of important tubulopathies in the pediatric ages(AU)


Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Osmolar Concentration , Diabetes Insipidus, Nephrogenic , Acute Kidney Injury , Clinical Laboratory Services , Hydrogen-Ion Concentration
17.
Rev. cuba. hematol. inmunol. hemoter ; 37(1): e1338, ene.-mar. 2021. tab, graf
Article in Spanish | LILACS, CUMED | ID: biblio-1251717

ABSTRACT

Introducción: Las hemoglobinopatías se consideran errores monogénicos hereditarios y están caracterizados por defectos en la molécula de hemoglobina. En Cuba, la detección prenatal de hemoglobinopatías se realiza a través de la electroforesis de hemoglobina para identificar parejas de alto riesgo. El programa brinda: asesoramiento genético, diagnóstico prenatal molecular e interrupciones selectivas de fetos afectados, a solicitud de las parejas. Objetivo: Determinar la frecuencia de hemoglobinopatías en mujeres embarazadas residentes en Cuba. Métodos: Se realizó un estudio descriptivo, retrospectivo y de corte transversal para determinar la frecuencia de hemoglobinopatías en 1 342 917 mujeres embarazadas captadas en el periodo 2009-2019. El método diagnóstico de la pesquisa fue la electroforesis de hemoglobina en geles de agarosa a pH alcalino. La confirmación se realizó por electroforesis de hemoglobina en gel de agarosa a pH ácido; ambos métodos mediante la tecnología HYDRASYS. Resultados: La frecuencia global de embarazadas con hemoglobinopatías fue de 3,5 por ciento. Se detectó hemoglobinopatías en 47 465 mujeres; 38 698 con variante S heterocigoto, 8 706 variantes de hemoglobina C y 158 de otras variantes. Se detectaron 44 283 esposos con hemoglobinopatías, 3 099 parejas de alto riesgo y se realizaron 2 689 diagnósticos prenatales moleculares. Se confirmaron 522 fetos afectados y 382 parejas solicitaron la interrupción del embarazo. El subprograma alcanzó 99,24 por ciento de cobertura en el país. Conclusión: La alta frecuencia de hemoglobinopatías en Cuba justifica la importancia de continuar el subprograma de detección de portadores para prevenir la aparición de las formas graves de la enfermedad(AU)


Introduction: Hemoglobinopathies are hereditary monogenic errors characterized by defects in the hemoglobin molecule. In Cuba, prenatal detection of hemoglobinopathies is performed by hemoglobin electrophoresis to identify high-risk couples. The program offers genetic counseling, prenatal molecular diagnosis and selective pregnancy termination in case of affected fetuses at the request of couples. Objective: Determine the frequency of hemoglobinopathies among pregnant women living in Cuba. Methods: A descriptive cross-sectional retrospective study was conducted to determine the frequency of hemoglobinopathies in 1 342 917 pregnant women recruited in the period 2009-2019. Screening was based on the diagnostic method of hemoglobin electrophoresis in alkaline pH agarose gels. Confirmation was performed with hemoglobin electrophoresis in acid pH agarose gel. Both methods used HYDRASYS technology. Results: Overall frequency of pregnant women with hemoglobinopathies was 3.5 percent. Hemoglobinopathies were detected in 47 465 women: 38 698 with variant S heterozygote, 8 706 with variants of hemoglobin C y 158 with other variants. 44 283 husbands with hemoglobinopathies and 3 099 high-risk couples were detected, and 2 689 prenatal molecular diagnostic tests were conducted. A total 522 affected fetuses were confirmed, and 382 couples requested pregnancy termination. The subprogram achieved 99.24 percent coverage in the country. Conclusion: The high frequency of hemoglobinopathies in Cuba justifies the importance of continuing the carrier detection subprogram to prevent the emergence of severe forms of the disease(AU)


Subject(s)
Humans , Female , Pregnancy , Prenatal Diagnosis , Family Characteristics , Electrophoresis , Genetic Counseling , Hemoglobinopathies , Hydrogen-Ion Concentration , Mass Screening , Retrospective Studies
18.
Electron J Biotechnol ; 49: 56-63, Jan. 2021. tab, ilus, graf
Article in English | LILACS | ID: biblio-1291900

ABSTRACT

BACKGROUND: 2R,3R-butanediol dehydrogenase (R-BDH) and other BDHs contribute to metabolism of 3R/3S-Acetoin (3R/3S-AC) and 2,3-butanediol (2,3-BD), which are important bulk chemicals used in different industries. R-BDH is responsible for oxidizing the hydroxyl group at their (R) configuration. Bacillus species is a promising producer of 3R/3S-AC and 2,3-BD. In this study, R-bdh gene encoding R-BDH from Bacillus sp. DL01 was isolated, expressed and identified. RESULTS: R-BDH exerted reducing activities towards Diacetyl (DA) and 3R/3S-AC using NADH, and oxidizing activities towards 2R,3R-BD and Meso-BD using NAD+ , while no activity was detected with 2S,3S-BD. The R-BDH showed its activity at a wide range of temperature (25 C to 65 C) and pH (5.0­8.0). The R-BDH activity was increased significantly by Cd2+ when DA, 3R/3S-AC, and Meso-BD were used as substrates, while Fe2+ enhanced the activity remarkably at 2R,3R-BD oxidation. Kinetic parameters of the R-BDH from Bacillus sp. DL01 showed the lowest Km, the highest Vmax, and the highest Kcat towards the racemic 3R/3S-AC substrate, also displayed low Km towards 2R,3R-BD and Meso-BD when compared with other reported R-BDHs. CONCLUSIONS: The R-BDH from Bacillus sp. DL01 was characterized as a novel R-BDH with high enantioselectivity for R-configuration. It considered NAD+ and Zn2+ dependant enzyme, with a significant affinity towards 3R/3S-AC, 2R,3R-BD, and Meso-BD substrates. Thus, R-BDH is providing an approach to regulate the production of 3R/3S-AC or 2,3-BD from Bacillus sp. DL01.


Subject(s)
Bacillus subtilis/enzymology , Alcohol Oxidoreductases/isolation & purification , Alcohol Oxidoreductases/metabolism , Temperature , Kinetics , Hydrogen-Ion Concentration , Acetoin
19.
REME rev. min. enferm ; 25: e1382, 2021. graf
Article in English, Portuguese | LILACS, BDENF | ID: biblio-1340533

ABSTRACT

RESUMO Objetivo: relatar casos de cateteres nasoenterais mal posicionados, após inserção às cegas à beira leito. Método: são apresentados três casos, os quais ocorreram em uma instituição hospitalar localizada no estado de São Paulo. Os cateteres foram inseridos pelo enfermeiro conforme protocolo institucional, para posicionamento em nível entérico. Ausculta epigástrica e mensuração do pH foram os métodos empregados na confirmação do posicionamento dos cateteres e posteriormente foi realizado o exame de Raios-X, considerado exame de referência para confirmar o posicionamento de cateteres nasoenterais. Resultados: dos três cateteres mal posicionados, dois estavam com a extremidade distal projetada para a junção esofagogástrica e uma não foi visibilizada, apesar de a ausculta ter sido positiva e os valores de pH terem sido superiores a seis. Conclusão: os resultados revelaram importantes limitações de ambos os métodos na verificação do posicionamento de cateteres nasoenterais que foram inseridos recentemente às cegas à beira leito.


RESUMEN Objetivo: reportar casos de catéteres nasoenterales mal posicionados tras inserción ciega a pie de cama. Método: se presentan tres casos, ocurridos en un hospital ubicado en el estado de São Paulo. Los catéteres fueron insertados por el enfermero según protocolo institucional, para su colocación a nivel entérico. La auscultación epigástrica y la medición del pH fueron los métodos utilizados para confirmar la posición de los catéteres y posteriormente se realizó el examen de rayos X, considerado un examen de referencia para confirmar la posición de los catéteres nasoenterales. Resultados: de los tres catéteres mal posicionados, dos tenían el extremo distal proyectado a la unión esofagogástrica y uno no se visualizaba, a pesar de que la auscultación era positiva y los valores de pH superiores a seis. Conclusión: los resultados revelaron importantes limitaciones de ambos métodos para verificar la posición de los catéteres nasoenterales recientemente insertados a ciegas a la cabecera de la cama.


ABSTRACT Objective: to report cases of poorly positioned nasoenteral catheters after blind insertion at the bedside. Method: three cases are presented, which occurred in a hospital located in the state of São Paulo. The catheters were inserted by the nurse according to institutional protocol, for placement at the enteric level. Epigastric auscultation and pH measurement were the methods used to confirm the positioning of the catheters and subsequently the X-Ray exam was performed, considered a reference exam to confirm the positioning of nasoenteral catheters. Results: of the three poorly positioned catheters, two were with the distal end projected to the esophagogastric junction and one was not visualized, despite the auscultation being positive and the pH values being higher than six. Conclusion: the results revealed important limitations of both methods in verifying the positioning of nasoenteral catheters that were recently inserted blindly at the bedside.


Subject(s)
Humans , Catheters , Patient Care , Intubation, Gastrointestinal/methods , X-Rays , Enteral Nutrition , Hydrogen-Ion Concentration
20.
Article in English | LILACS, BBO | ID: biblio-1143394

ABSTRACT

ABSTRACT Objective: To analyze the ability of saliva in controlling the growth and the biofilm formation of Streptococcus mutans (S. mutans) as well as the effect of histatin-5 anti-biofilm relate to pH and saliva viscosity. Material and Methods: The S. mutans biofilm assayed by crystal violet 1% and its growth measured by spectrophotometer. The saliva viscosity was analyzed by viscometer, and pH of saliva was measured by pH meter. Results: Based on the optical density values, growth of S. mutans in saliva ranged <300 CFU/mL (0.1 nm) at concentrations of 25%, 12.5% and 6.25% for 24 hours. Whereas at the 48 h and 72 h period of incubation shown an increase in growth of S. mutans ranged 300-600 CFU/mL (0.2-0.36 nm). The inhibitory biofilm formation of S. mutans in saliva was significantly higher at concentrations of 12.5% and 6.25% at 24 h incubation times on a moderate scale, whereas the histatin-5 was effective to inhibit S. mutans biofilm on the 50 and 25 ppm. The saliva possessed a higher inhibitory of biofilm S. mutans than histatin-5 and good level viscosity (0.91-0.92 cP). Conclusion: The saliva was able to control the growth of S. mutans, and histatin-5 can inhibit the biofilm formation S. mutans. Furthermore, the saliva was also able to respond to the pH change with good viscosity of saliva.


Subject(s)
Humans , Male , Female , Child , Saliva/microbiology , Biofilms , Viridans Streptococci , Histatins , Hydrogen-Ion Concentration , Spectrophotometry/instrumentation , Streptococcus mutans , Viscosity , Analysis of Variance , Statistics, Nonparametric , Indonesia/epidemiology
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