Phytochemical screening of Cordia glabrata (MART.) A.DC. extracts and its potential antioxidant, photoprotective, antimicrobial and antiviral activities / Triagem fitoquímica de extratos de Cordia glabrata (MART.) A.DC. e sua potencial atividade antioxidante, fotoprotetora, antimicrobiana e antiviral

Species of the genus Cordia have shown biological activities, such as anti-inflammatory, analgesic, antioxidant, antiviral, and antifungal activities. The species Cordia glabrata (MART) A.DC. Has no information concerning its phytochemical profile and possible biological activities. Thus, this study aimed to evaluate this profile in ethanolic extracts of young, adult and senescent leaves, as well as their antioxidant, photoprotective, antimicrobial, and virucidal potentials. Phytochemical analysis was performed by TLC (thin-layer chromatography) and showed the presence of flavonoids, tannins, and terpenes. The evaluation by UPLC-MS/MS (Ultra performance liquid chromatography - tandem mass spectrometer) evidenced the presence of caffeic (3.89 mgL-¹), p-cumaric (6.13 mgL-¹), and ferulic (0.58 mgL-¹) acids, whilst, in GC/MS (Gas chromatography–mass spectrometry) analysis there was a greater amount of palmitic (51.17%), stearic (20.34%), linoleic (9.62%), and miristic (8.16%) fatty acids. The DPPH (2,2-Diphenyl-1-picrylhydrazyl) and ABTS+ (2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) radicals were used to verify the potential antioxidant activity, observing a better activity for the leaf extract in the adult phenological stage: 54.63 ± 1.06 µgmL-¹ (DPPH) and 44.21 ± 1.69 mM (ABTS). The potential photoprotective activity of the extracts was determined by spectrophotometry and the in vitro values of SPF (Sun Protection Factor) in young and adult leaves (5.47 and 5.41, respectively) showed values close to the minimum SPF of 6.0 required by ANVISA (Brazilian Health Regulatory Agency). It was not observed an antimicrobial activity for Staphylococcus aureus with a minimum inhibitory concentration of 2000 µgmL-¹, however the anti-herpetic assay against the Herpes simplex virus type 2 (HSV-2) showed a potent virucidal activity at the tested concentrations with CV50 value <0.195 µgmL-¹ and a Selectivity Index (SI = CC50 / CV50) greater than 448. The results [...].

Espécies do gênero Cordia apresentam atividades biológicas, como anti-inflamatória, analgésica, antioxidante, antiviral e antifúngica. Para a espécie Cordia glabrata (MART) A.DC., ainda não existem informações sobre seu perfil fitoquímico e possíveis atividades biológicas, deste modo, o presente estudo teve como objetivo avaliar este perfil em extratos etanólicos de folhas jovens, adultas e senescentes, bem como o potencial antioxidante, fotoprotetor, antimicrobiano e virucida. A análise fitoquímica foi realizada por CCD (Cromatografia em Camada Delgada), mostrando a presença de flavonóides, taninos e terpenos. Na avaliação por CLAE EM/EM (Cromatografia Líquida de Ultra Eficiência acoplada a Espectrometria de Massas) foi evidenciado a presença dos ácidos caféico (3,89 mgL-¹), p-cumárico (6,13 mgL-¹) e ferúlico (0,58 mgL-¹), paralelamente, na CG/EM (Cromatografia Gasosa acoplada a Espectrometria de Massas) verificou-se maior quantidade dos ácidos graxos palmítico (51,17%), esteárico (20,34%), linoléico (9,62%) e mirístico (8,16%). Os radicais DPPH (2,2-Difenil-1-picrilhidrazil) e ABTS+ (2′-Azino-bis (ácido 3-etilbenzotiazolina-6-sulfônico)) foram utilizados para verificar o potencial antioxidante, observando se uma atividade superior para o extrato da folha em sua fase fenológica adulta: 54,63 ± 1,06 µgmL-¹ (DPPH) e 44,21 ± 1,69 mM (ABTS+). A potencial atividade fotoprotetora dos extratos foi determinada espectrofotometricamente e os valores in vitro de FPS (Fator de Proteção Solar) em folhas jovens e adultas (5,47 e 5,41 respectivamente) apresentaram valores próximos ao FPS mínimo de 6,0 exigido pela ANVISA (Agência Nacional de Vigilância Sanitária). Não foi observada atividade antimicrobiana para Staphylococcus aureus sendo a concentração inibitória mínima de 2000 µgmL-¹, no entanto o ensaio anti-herpético contra o vírus Herpes simplex tipo 2 (HSV-2) mostrou uma potente atividade virucida nas concentrações testadas [...].

Gestión de riesgos relativo al cambio de campaña productiva durante la etapa de desarrollo tecnológico en una instalación multiproducto / Risk-management related to the productive campaign change during technological development stage in a multi-product facility

Los nuevos paradigmas de la industria farmacéutica en el siglo XXI incorporan el uso de las técnicas de gestión de la calidad, necesarias en el cumplimiento de las buenas prácticas de fabricación en el sector biotecnológico. En este caso de estudio, se aplicó la gestión de riesgo en el cambio de campaña entre los ingredientes farmacéuticos activos de futuros candidatos inmunoterapeúticos contra el cáncer, en la etapa de desarrollo tecnológico en una instalación multiproducto certificada. Las causas potenciales de mayor influencia en las fallas son: la calificación del personal de la Dirección de Desarrollo Tecnológico en los procedimientos patrones de operación de la planta, la mezcla entre los componentes y materiales no dedicados utilizados en el proceso de purificación cromatográfica, la documentación en elaboración o aprobación y el establecimiento de las técnicas analíticas en función de la etapa del proyecto. Como resultado se proponen acciones que minimizan los riesgos de la contaminación cruzada y hacen viable un adecuado cambio de campaña entre la fabricación de los inmunoterapéuticos, durante el desarrollo tecnológico en una instalación multiproducto de la industria biotecnológica(AU)

The new paradigms of the pharmaceutical industry in the 21st century introduce the use of modern quality management techniques to comply with good manufacturing practices in the biotechnological area. In this paper, it was applied the risk management for the campaign change among the process for obtaining the active pharmaceutical ingredients to future immunotherapeutic candidates at the technological development stage in a certified multi-product facility. Particularly, the training for the development personnel in the standard operating procedures of the facility, the mixture between the components and non-dedicated materials used in the chromatographic purification process, the documentation in the preparation or approval, and the establishment of analytical techniques depending on the stage of the project are the potential causes of greater influence. As a result, actions are proposed to minimize risks and carry out an adequate campaign change feasible between the manufacture of immunotherapeutics during the technological development stage in a biotechnological multi-product facility(AU)

Aplicación del Análisis de Componentes Principales en el proceso de purificación de un biofármaco / Application of Principal Component Analysis in the purification process of a biopharmaceutical

En este trabajo se presenta la aplicación del Análisis de Componentes Principales, mediante el programa THE UNSCRAMBLER versión 8.0, a los datos registrados en un período de 2 años en la etapa de purificación de una planta de producción de Eritropoyetina Humana Recombinante que está basada en varios pasos cromatográficos, de forma similar a los procesos de purificación de proteínas recombinantes que se utilizan como vacunas preventivas o terapéuticas. Se logró reducir dimensionalidad al obtenerse dos componentes principales que explican el 81 por ciento de la varianza de 18 variables originales relacionadas con cuatro pasos cromatográficos. Como resultado se llegó a definir cuáles son las variables que mayor aporte tienen a la variabilidad del proceso en la etapa de purificación, permitiendo extraer información útil para lograr un mayor entendimiento del proceso y enriquecer las estrategias de control en la planta. Dichos resultados corroboraron experiencias prácticas de especialistas de la planta y permitieron dar recomendaciones a considerar en el plan de verificación continuada del proceso como proponer cinco variables como controles de proceso y tener en cuenta que el rendimiento del segundo paso cromatográfico es el más influyente de los rendimientos considerados en la variabilidad(AU)

This paper presents the application of the Principal Component Analysis, using the program THE UNSCRAMBLER version 8.0, to the data recorded during two years in the purification stage of a Recombinant Human Erythropoietin plant that is based on several chromatographic steps, similar to the purification process of recombinant proteins that are used as preventive or therapeutic vaccines. Dimensionality was reduced by obtaining two main components that explain 81 percent of the variance of 18 original variables related to four chromatographic steps. As a result, it was possible to define which variables have the greatest contribution to the variability of the process in the purification stage, allowing to extract useful information to achieve a greater understanding of the process and enrich the control strategies in the plant. These results corroborated practical experiences of plant specialists and allowed for recommendations to be considered in the continuous verification plan of the process, such as proposing three variables as process controls and taking into account that the performance of the second step is the most influential of the performances considered in the variability(AU)

Current Approach in Radiochemical Quality Control of the 99mTc-Radiopharmaceuticals: a mini-review

Abstract To present optimized chromatographic systems for radiochemical purity (RCP) evaluation of 99mTc-eluate and 99mTc-radiopharmaceuticals, as well as to assess doses calibrator reliability for routine purposes in hospital radiopharmacies. RCP was determined by different systems and radioactivity was quantified by TLC-scanner, doses calibrator and gamma-counter. Suitable and optimized systems were presented for RCP analyses. No significant differences were observed between radioactivity counting devices and, thus, doses calibrator showed reliability for RCP determination in hospital radiopharmacies.

Bioprospección de la actividad antimicrobiana de los extractos de los hongos endófitos de Bauhinia guianensis / Bioprospecting of antimicrobial activity of extracts of endophytic fungi from Bauhinia guianensis

Antibiotic resistance results in higher medical costs, prolonged hospital stays and increased mortality and is rising to dangerously high levels in all parts of the world. Therefore, this study aims to search for new antimicrobial agents through bioprospecting of extracts of endophytic fungi from Bauhinia guianensis, a typical Amazonian plant used in combating infections. Seventeen (17) fungi were isolated and as result the methanolic extract of the fungus Exserohilum rostratum showed good activity against the bacteria tested. The polyketide monocerin was isolated by the chromatographic technique, identified by NMR and MS, showing broad antimicrobial spectrum.

La resistencia a los antibióticos conduce a mayores costos médicos, hospitalizaciones prolongadas e incremento de la mortalidad, y está aumentando a niveles peligrosamente altos en todas partes del mundo. Este estudio tuvo como objetivo la búsqueda de nuevos agentes antimicrobianos a través de la bioprospección de extractos de hongos endófitos de Bauhinia guianensis, una planta amazónica típica, utilizada en la lucha contra problemas infecciosos. Fueron aislados 17 hongos; el extracto metanólico del hongo Exserohilum rostratum mostró buena actividad contra las bacterias probadas. Se aisló monocerina policétido por la técnica de cromatografía; este compuesto fue identificado por RM y EM, y mostró un amplio espectro antimicrobiano.

Preliminary analysis of Brazilian Amazonian plant extracts using bench-top assays in a through-put basis aiming the identification of potential antioxidant natural product

Plants are a source of compounds that are used for the treatment of human and veterinary diseases. Brazil is one of the richest countries in the world in terms of biodiversity. The present study evaluated extracts using thin-layer chromatography to identify antioxidant activity and determine the presence of groups of compounds, in a large-scale basis. A total of 1,260 aqueous and organic extracts were obtained from plants that were collected in the Amazon and Atlantic rain forests. Thin-layer chromatography was performed to evaluate the presence of alkaloids, anthraquinones, cardioactive glicosides, terpenes, and phenolics and determine antioxidant and radical scavenging activity using the following reagents: Dragendorff's reagent, KOH, Kedde's reagent, sulphuric acid, NP reagent, ß-carotene, and 2,2-diphenyl-1-picrylhydrazyl (DPPH). Of the 1,260 extracts, 837 (66.43%) presented a ß-carotene/bleaching response, and 1,205 (95.63%) presented a radical scavenging response. Alkaloids were found in 203 extracts (16.11%). Anthraquinones were found in 14 extracts (1.11%), cardenolides were found in eight extracts (0.63%). The present findings shows the importance of the Brazilian Amazon plants as sources of antioxidant and radical scavenging active compounds.(AU)

Indole acetic acid and ACC deaminase from endophytic bacteria improves the growth of Solarium lycopersicum

Background: Endophytic bacteria are ubiquitous in all plant species contributing in host plant's nutrient uptake and helping the host to improve its growth. Moringa peregrina which is a medicinal plant, growing in arid region of Arabia, was assessed for the presence of endophytic bacterial strains. Results: PCR amplification and sequencing of 16S rRNA of bacterial endophytes revealed the 5 endophytic bacteria, in which 2 strains were from Sphingomonas sp.; 2 strains from Bacillus sp. and 1 from Methylobacterium genus. Among the endophytic bacterial strains, a strain of Bacillus subtilis LK14 has shown significant prospects in phosphate solubilization (clearing zone of 56.71 mm after 5 d), ACC deaminase (448.3 ± 2.91 nM α-ketobutyrate mg-1 h-1) and acid phosphatase activity (8.4 ± 1.2 nM mg-1 min-1). The endophytic bacteria were also assessed for their potential to produce indole-3-acetic acid (IAA). Among isolated strains, the initial spectrophotometry analysis showed significantly higher IAA production by Bacillus subtilis LK14. The diurnal production of IAA was quantified using multiple reactions monitoring method in UPLC/MS-MS. The analysis showed that LK14 produced the highest (8.7 uM) IAA on 14th d of growth. Looking at LK14 potentials, it was applied to Solanum lycopersicum, where it significantly increased the shoot and root biomass and chlorophyll (a and b) contents as compared to control plants. Conclusion: The study concludes that using endophytic bacterial strains can be bio-prospective for plant growth promotion, which might be an ideal strategy for improving growth of crops in marginal lands.

Estudio de la capacidad antioxidante in vitro de Phaseolus vulgaris L. (frijol) mediante extracción asistida por microondas / Study of in vitro antioxidant capacity of Phaseolus vulgaris L. (frijol) by microwave assisted extraction

Introducción: Phaseolus vulgaris L. (frijol) es una fuente nutricional importante en Colombia, que aporta un gran contenido de sustancias bioactivas con potencial benéfico para la salud, tales como polifenoles, entre otras, que contribuyen de manera sinérgica con sus propiedades terapéuticas, y que pueden tener un efecto positivo contra algunas patologías. Objetivos: evaluar el método de extracción asistido por microondas como método alternativo para estudiar la capacidad antioxidante in vitro en ocho variedades de P. vulgaris L. cultivadas en Colombia. Métodos: semillas sin piel de P. vulgaris, deshidratadas y maceradas se sometieron a extracción asistida por microondas y extracción sólido-líquido; el contenido de fenoles se evaluó por el método de Folin-Ciocalteu y el potencial antioxidante in vitro se evaluó con base en los métodos del radical estable catión radical difenil-picrilhidrazilo y el radical catión 2,2´-azino-bis(ácido 3-etilbenzotiazolin-6-sulfunico). Resultados: el método de extracción asistido por microondas realizada en horno microondas convencional fue más eficiente respecto a la convencional ya que disminuyó la cantidad de solvente, de muestra empleada y los tiempos de extracción. Los extractos obtenidos por extracción asistida por microondas en horno microndas convencional presentaron un contenido de fenoles entre 29,36 y 60,61 g EAG/L, mientras que el método extracción sólido-líquido, estuvo entre 32,75 y 113,27 g EAG/L. El efecto anti-radicalario fue similar entre los extractos evaluados. Conclusiones: todos los extractos presentaron buena capacidad protectora contra radicales libres, y la técnica de extracción asistida por microondas en horno microndas convencional puede ser usada como método alterno para una valoración rápida, eficiente y eficaz del contenido de sustancias bioactivas en diferentes matrices, se presentó mínimas diferencias entre los resultados obtenidos, comparados con las metodologías de extracción asistida por microondas establecidas antes(AU)

Introduction: Phaseolus vulgaris L. is a representative crop of nutrient and economic importance in Colombia. Additionally, P. vulgaris is considered as a natural source of bioactives compounds, such as polyphenols, which have been associated with valuable effects on health. Objetives: to evaluate the microwave extraction assisted technique as an alternative methodology to study the antioxidant capacity of eight varieties of P. vulgaris cultivated in Colombia. Methods: dehydrated and powered seeds of P. vulgaris was subjected to microwave assisted extraction and solid-liquid extraction and. Total phenolis content was determined by Folin-Cicoulteau method and the potential antioxidant was evaluated using diphenyl-picrylhydrazyl radical stable and 2,2-Azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical cation assays. Results: microwave assisted extraction-Household microwave oven technique was more efficient and versatile than SLE method. The extracts obtained microwave assisted extraction-Household microwave oven methodology showed polyphenol content ranged between entre 29,36 and 60,61 g EGA/L, but SLE was over 32,75 and 113,27 g EAG/L. Conclusions: all extracts showed a considerable antioxidant potential against free radical, and microwave assisted extraction-Household microwave oven method could be used as an alternative method for fast, efficient and effective evaluation of the content of polyphenol in various matrices, with minimal differences comparing to established microwave assisted extraction techniques(AU)

Alternative chromatographic system for the quality control of lipophilic technetium-99m radiopharmaceuticals such as [99mTc(MIBI)6]+

Knowledge of the radiochemical purity of radiopharmaceuticals is mandatory and can be evaluated by several methods and techniques. Planar chromatography is the technique normally employed in nuclear medicine since it is simple, rapid and usually of low cost. There is no standard system for the chromatographic technique, but price, separation efficiency and short time for execution must be considered. We have studied an alternative system using common chromatographic stationary phase and alcohol or alcohol:chloroform mixtures as the mobile phase, using the lipophilic radiopharmaceutical [99mTc(MIBI)6]+ as a model. Whatman 1 modified phase paper and absolute ethanol, Whatman 1 paper and methanol:chloroform (25:75), Whatman 3MM paper and ethanol:chloroform (25:75), and the more expensive ITLC-SG and 1-propanol:chloroform (10:90) were suitable systems for the direct determination of radiochemical purity of [99mTc(MIBI)6]+ since impurities such as99mTc-reduced-hydrolyzed (RH),99mTcO4- and [99mTc(cysteine)2]-complex were completely separated from the radiopharmaceutical, which moved toward the front of chromatographic systems while impurities were retained at the origin. The time required for analysis was 4 to 15 min, which is appropriate for nuclear medicine routines.

Caracterización bioquímica y biológica del veneno de la serpiente Neotropical Macagua (Bothrops Colombiensis) de la región de Barlovento, estado Miranda, Venezuela / Biological and biochemical characterization of the Neotropical Macagua (Bothrops colombiensis) snake venom from Barlovento region, Miranda state, Venezuela

Introducción: el veneno de B. colombiensis no es solamente un elemento tóxico; en su composición existen múltiples componentes, que tienen un gran potencial terapéutico, principalmente en el tratamiento de patologías de la trombosis y la coagulación. Objetivos: estudiar una mezcla de venenos de Bothrops colombiensis de una ubicacion geográfica de Venezuela, a fin de hacer un barrido de sus actividades hemostáticas, que permitirá posteriormente purificar y caracterizar moléculas con actividad antitrombótica y anticoagulantes, entre otras, con potencial terapéutico. Métodos: el veneno a estudiar, es una mezcla de ellos obtenidos de serpientes provenientes de la Región de Barlovento, estado Miranda, Venezuela. Se caracterizó bioquímicamente por cromatografias de exclusión molecular, cromatografía de fase reversa C18 y por electroforesis a través de SDS­PAGE; y biológicamente por medio de actividades relacionadas con la hemostasia. Se analizaron los perfiles en relación a las actividades fibrinolítica, proteolítica sobre polvo azul y cadena ß de insulina, procoagulante, hemorrágica y letal. Resultados: la actividad hemorrágica, definida como la Dosis Hemorrágica Mínima fue de 8,7 mg/kg. La letalidad, definida como la Dosis Letal cincuenta fue 8,7 mg/kg. El veneno presentó actividad procoagulante y fibrinolítica. Las fracciones mostraron actividad fibrinolítica y proteolítica sobre polvo azul de ocultamiento y sobre la cadena ß de insulina. Conclusiones: las características biológicas de los componentes de este veneno le confieren un enorme potencial terapéutico, ya que contiene una alta actividad fibrinolítica y anticoagulante. Estos compuestos una vez purificados y caracterizados podrían explorarse como coadyuvantes en procesos trombolíticos, dado que disuelven coágulos de fibrina y degradan fibrinógeno, evitando episodios de retrombosis(AU)

Introduction: This paper is a screening of multiple toxic activities, of which some will be potentially useful for the management of coagulation pathologies. Objetives: A pool of Bothrops colombiensis venoms from a specific geographical location was studied, in order to carry out a hemostatic activities screening, allowing then to purify and characterise molecules with antithrombotic and anticoagulant activity, among others, which could have therapeutic potential. Methods: The venom was chromatographically by molecular exclusion and reverse phase C18 and SDS -PAGE characterized; its hemostatic activity was also established. Snakes were from the region of Barlovento, Miranda state, Venezuela. Profiles of fibrinolytic, proteolytic, procoagulant, hemorrhagic and lethal activities were analyzed. Hemorrhagic activity was 8.7 mg/kg. The LD50 was 8.7 mg/kg. The venom showed strongly procoagulant activity. Both, crude venom as fractions showed high fibrinolytic activity. The majority of the eluted fractions showed significant proteolytic activity in azure blue powder and on ß chain of insulin. Conclusions: The biological characteristics of the components of this venom confer enormous therapeutic potential because they contain a high fibrinolytic and anticoagulant activity. Most of these proteinases, once purified and characterized, could be explored as thrombolytic agents given that dissolves fibrin clots or prevent their formation(AU)

Xylooligosaccharides: an economical prebiotic from agroresidues and their health benefits.

Oligosaccharides and dietary fibres are non-digestible food ingredients that preferentially stimulate the growth of prebiotic Bifidobacterium and other lactic acid bacteria in the gastro-intestinal tract. Xylooligosaccharides (XOS) provide a plethora of health benefits and can be incorporated into several functional foods. In the recent times, there has been an over emphasis on the microbial conversion of agroresidues into various value added products. Xylan, the major hemicellulosic component of lignocellulosic materials (LCMs), represents an important structural component of plant biomass in agricultural residues and could be a potent bioresource for XOS. On an industrial scale, XOS can be produced by chemical, enzymatic or chemo-enzymatic hydrolysis of LCMs. Chemical methods generate XOS with a broad degree of polymerization (DP), while enzymatic processes will be beneficial for the manufacture of food grade and pharmaceutically important XOS. Xylooligomers exert several health benefits, and therefore, have been considered to provide relief from several ailments. This review provides a brief on production, purification and structural characterization of XOS and their health benefits.

Análise in vitro de um dispositivo polimérico como alternativa para o uso de antimicrobiano sistêmico em Odontologia / In vitro analysis of a polymeric device as an alternative for systemic antibiotics in Dentistry

A administração indiscriminada de antimicrobianos sistêmicos tem como principais efeitos indesejáveis a seleção antimicrobiana, hipersensibilidade, comprometimento gastrointestinal e toxicidade. A busca por uma alternativa à terapêutica antimicrobiana sistêmica em Odontologia através do uso de um material biodegradável de aplicação local pode apresentar inúmeras vantagens. As características estruturais, de citocompatibilidade e facilidade de fabricação do polímero sintético ácido poli-L-lactídeo (PLLA) permitem que este seja um carreador de fármacos como amoxicilina (AM), azitromicina (AZ), clindamicina (CL) ou metronidazol (ME) mantendo concentrações inibitórias constantes e por tempo prolongado, sendo capazes de prevenir a colonização dos principais patógenos orais. Objetivo: Avaliar e comparar o comportamento de filmes ou malhas de PLLA associados aos quatro antimicrobianos mais utilizados em Odontologia como uma alternativa local. Metodologia: 180 (N) discos poliméricos com 15 ou 6 mm de diâmetro foram preparados em associação a 20% do antimicrobiano amoxicilina, azitromicina, clindamicina ou metronidazol sendo classificados como grupo F (filme) e M (malha). Foram confeccionados segundo os métodos de deposição e eletrofiação (fibras) respectivamente. Todos os discos foram armazenados em solução tampão (pH 5 ou 7.4) e alíquotas foram coletadas e analisadas por cromatografia líquida de alta performace (HPLC) em 8, 24, 48, 72, 96, 120, 144 e 168 horas. As espécimes foram pesadas após 3 e 6 meses de armazenamento nas soluções tampões para análise de degradação. Para a análise de citotoxicidade, os materiais foram cultivados com fibroblastos humanos por 24h, 48h e 72h e analisados por ensaio de MTT. A capacidade antimicrobiana dos discos foi determinada em cultura de P.gingivalis e S.pyogenes. Para o controle estrutural foram realizadas fotografias digitais e MEV dos espécimes controle, das interfaces (criofratura) e das espécimes degradadas. Resultados: A liberação farmacológica para os antimicrobianos na ordem pH levemente básico (7.4) e ácido (5.0) foi respectivamente: ME 70.03% (F) e 100% (M); 88,01% (F) e 19,4% (M). Para AM 38,73% (F) e 18,63% (M); 61,44% (F) e 47,93% (M). Para AZ 32,53% (F) e 82,85% (M); 46,78% (F) e 73,15% (M). Para CL 68,42% (F) e 81,10% (M); 76,47% (F) e 72,76% (M). A análise antimicrobiana demonstrou capacidade inibitória para S.pyogenes e P.gingivalis para todos os materiais testados, não havendo diferença significativa entre filme e malha dentro de cada grupo (p>0.05). A reação de citotoxicidade por MTT comprovou que os biomateriais testados são compatíveis com fibroblastos humanos e mais citocompatíveis que o controle PLLA, controle de vida e morte (p<0.05). As malhas demonstraram favorecimento do crescimento celular principalmente em 24 e 48 horas. A MEV demonstra um filme com superfície rugosa e malha com fibras e poros mimetizando a matriz extracelular. Após criofratura a MEV da interface comprovou incorporação do fármaco ao filme e malha, exceto para o ME, com cristais externos ao polímero. Após a degradação, os filmes de amoxicilina apresentaram maior degradação que PLLA no pH 5.0 (p=0.007) e pH 7.4 (p=0.046). Já para as malhas a azitromicina apresentou maior degradação que PLLA no pH 7.4 (p=0.031). Conclusão: O PLLA é um polímero cuja associação aos antimicrobianos utilizados mostrou-se segura, citocompatível e promissora na liberação de doses inibitórias contra os microrganismos P.gingivalis e S. pyogenes. A liberação farmacológica foi influenciada pela característica química do fármaco, apresentação do polímero (filme e malha) e pH da solução de armazenamento. Este estudo comprovou ser possível através de uma terapêutica medicamentosa local controlar ou prevenir infecções localizadas...

Indiscriminate administration of systemic antimicrobial has undesirable effects such as antimicrobial selection, hypersensitivity, gastrointestinal commitment and toxicity. For an alternative to systemic antimicrobial therapy in Dentistry, use a biodegradable material of local application can present numerous advantages. The structural characteristics, cytocompatibility and ease of fabrication of the synthetic polymer poly-L- lactide acid (PLLA) enable this to be a carrier biomaterial. When associated with antimicrobials as amoxicillin (AM), azithromycin (AZ), clindamycin (CL) or metronidazole (ME) it can maintain constant the inhibitory concentrations for a long time, being able to prevent colonization of the main oral pathogens. Objective: To evaluate and compare the behavior of PLLA associated with the most useful antimicrobials in Dentistry as an alternative for prevention and treatment of infections. Methodology: 180 (N) polymer discs with 15 or 6 mm diameter were prepared in association with the antimicrobial concentration of 20% amoxicillin, metronidazole, clindamycin or azithromycin being classified as Group F (film) and M (mesh). They were made using the methods of deposition and electrospinning (nanofibers) respectively. All discs were stored in buffer solutions (pH 5 or 7.4) and aliquots were collected and analyzed by high performance chromatography (HPLC) on 8, 24, 48, 72, 96, 120 , 144 and 168 hours. Cytotoxicity of human fibroblasts was tested after 24h, 48h and 72h by the MTT reaction. The antimicrobial capacity of the disks was determined against P. gingivalis and S. pyogenes cultures. The specimens were weighed after 3 and 6 months of storage for degradation analysis. Specimens were also carried out by digital photos for structural control. SEM was used to control interfaces (freeze-fracture) and degradation description. Results: The drug release for antimicrobials in order slightly basic pH (7.4) and acid ( 5.0 ) was respectively...

Análise in vitro de um dispositivo polimérico como alternativa para o uso de antimicrobiano sistêmico em Odontologia / In vitro analysis of a polymeric device as an alternative for systemic antibiotics in Dentistry

A administração indiscriminada de antimicrobianos sistêmicos tem como principais efeitos indesejáveis a seleção antimicrobiana, hipersensibilidade, comprometimento gastrointestinal e toxicidade. A busca por uma alternativa à terapêutica antimicrobiana sistêmica em Odontologia através do uso de um material biodegradável de aplicação local pode apresentar inúmeras vantagens. As características estruturais, de citocompatibilidade e facilidade de fabricação do polímero sintético ácido poli-L-lactídeo (PLLA) permitem que este seja um carreador de fármacos como amoxicilina (AM), azitromicina (AZ), clindamicina (CL) ou metronidazol (ME) mantendo concentrações inibitórias constantes e por tempo prolongado, sendo capazes de prevenir a colonização dos principais patógenos orais. Objetivo: Avaliar e comparar o comportamento de filmes ou malhas de PLLA associados aos quatro antimicrobianos mais utilizados em Odontologia como uma alternativa local. Metodologia: 180 (N) discos poliméricos com 15 ou 6 mm de diâmetro foram preparados em associação a 20% do antimicrobiano amoxicilina, azitromicina, clindamicina ou metronidazol sendo classificados como grupo F (filme) e M (malha). Foram confeccionados segundo os métodos de deposição e eletrofiação (fibras) respectivamente. Todos os discos foram armazenados em solução tampão (pH 5 ou 7.4) e alíquotas foram coletadas e analisadas por cromatografia líquida de alta performace (HPLC) em 8, 24, 48, 72, 96, 120, 144 e 168 horas. As espécimes foram pesadas após 3 e 6 meses de armazenamento nas soluções tampões para análise de degradação. Para a análise de citotoxicidade, os materiais foram cultivados com fibroblastos humanos por 24h, 48h e 72h e analisados por ensaio de MTT. A capacidade antimicrobiana dos discos foi determinada em cultura de P.gingivalis e S.pyogenes.

Para o controle estrutural foram realizadas fotografias digitais e MEV dos espécimes controle, das interfaces (criofratura) e das espécimes degradadas. Resultados: A liberação farmacológica para os antimicrobianos na ordem pH levemente básico (7.4) e ácido (5.0) foi respectivamente: ME 70.03% (F) e 100% (M); 88,01% (F) e 19,4% (M). Para AM 38,73% (F) e 18,63% (M); 61,44% (F) e 47,93% (M). Para AZ 32,53% (F) e 82,85% (M); 46,78% (F) e 73,15% (M). Para CL 68,42% (F) e 81,10% (M); 76,47% (F) e 72,76% (M). A análise antimicrobiana demonstrou capacidade inibitória para S.pyogenes e P.gingivalis para todos os materiais testados, não havendo diferença significativa entre filme e malha dentro de cada grupo (p>0.05). A reação de citotoxicidade por MTT comprovou que os biomateriais testados são compatíveis com fibroblastos humanos e mais citocompatíveis que o controle PLLA, controle de vida e morte (p<0.05). As malhas demonstraram favorecimento do crescimento celular principalmente em 24 e 48 horas. A MEV demonstra um filme com superfície rugosa e malha com fibras e poros mimetizando a matriz extracelular. Após criofratura a MEV da interface comprovou incorporação do fármaco ao filme e malha, exceto para o ME, com cristais externos ao polímero. Após a degradação, os filmes de amoxicilina apresentaram maior degradação que PLLA no pH 5.0 (p=0.007) e pH 7.4 (p=0.046). Já para as malhas a azitromicina apresentou maior degradação que PLLA no pH 7.4 (p=0.031). Conclusão: O PLLA é um polímero cuja associação aos antimicrobianos utilizados mostrou-se segura, citocompatível e promissora na liberação de doses inibitórias contra os microrganismos P.gingivalis e S. pyogenes. A liberação farmacológica foi influenciada pela característica química do fármaco, apresentação do polímero (filme e malha) e pH da solução de armazenamento. Este estudo comprovou ser possível através de uma terapêutica medicamentosa local controlar ou prevenir infecções localizadas...

Indiscriminate administration of systemic antimicrobial has undesirable effects such as antimicrobial selection, hypersensitivity, gastrointestinal commitment and toxicity. For an alternative to systemic antimicrobial therapy in Dentistry, use a biodegradable material of local application can present numerous advantages. The structural characteristics, cytocompatibility and ease of fabrication of the synthetic polymer poly-L- lactide acid (PLLA) enable this to be a carrier biomaterial. When associated with antimicrobials as amoxicillin (AM), azithromycin (AZ), clindamycin (CL) or metronidazole (ME) it can maintain constant the inhibitory concentrations for a long time, being able to prevent colonization of the main oral pathogens. Objective: To evaluate and compare the behavior of PLLA associated with the most useful antimicrobials in Dentistry as an alternative for prevention and treatment of infections. Methodology: 180 (N) polymer discs with 15 or 6 mm diameter were prepared in association with the antimicrobial concentration of 20% amoxicillin, metronidazole, clindamycin or azithromycin being classified as Group F (film) and M (mesh). They were made using the methods of deposition and electrospinning (nanofibers) respectively. All discs were stored in buffer solutions (pH 5 or 7.4) and aliquots were collected and analyzed by high performance chromatography (HPLC) on 8, 24, 48, 72, 96, 120 , 144 and 168 hours. Cytotoxicity of human fibroblasts was tested after 24h, 48h and 72h by the MTT reaction. The antimicrobial capacity of the disks was determined against P. gingivalis and S. pyogenes cultures. The specimens were weighed after 3 and 6 months of storage for degradation analysis.

Specimens were also carried out by digital photos for structural control. SEM was used to control interfaces (freeze-fracture) and degradation description. Results: The drug release for antimicrobials in order slightly basic pH (7.4) and acid ( 5.0 ) was respectively ...

Determinação da presença de 5-FU na saliva de hamsters que receberam o quimioterápico pela técnica de Cromatografia Líquida de Alta Eficiência / Determining the presence of 5-FU in the saliva of the hamsters that received chemotherapy by liquid chromatography of high efficiency

Vários métodos de análise para o ensaio do quimioterápico 5-Fluorouracil (5-FU) em fluidos biológicos de humanos e animais, foram previamente relatados. Considerando que a administração do 5-FU altera de alguma maneira a morfologia e função das glândulas salivares, e que a presença do quimioterápico na mucosa oral pode levar a algumas complicações orais, este trabalho teve como objetivo de determinar a presença de 5-FU na saliva de hamsters que receberam o quimioterápico pela técnica de Cromatografia Líquida de Alta Eficiência (CLAE), uma vez que este modelo animal é usado nos estudos com mucosite oral e hipofunção glandular, induzidas por 5-FU. Doze animais foram divididos em 4 grupos: CP e CPI, onde os animais receberam intraperitonealmente pilocarpina (CP) ou pilocarpina + isoproterenol (CPI) e o veículo do quimioterápico, e os grupos QP e QPI, onde os animais receberam, respectivamente, os mesmos secretagogos listados acima e o quimioterápico 5-FU. Após a administração do secretagogo, foi coletada a saliva de todos os animais, por um período de 60 min. Em seguida, a saliva foi congelada a -80 ?C para posterior determinação do quimioterápico por CLAE. Após análise dos cromatogramas, e com base nos resultados obtidos, foi possível identificar a presença do 5-FU nas amostras de saliva de hamsters que receberam o quimioterápico via intraperitoneal pela técnica da CLAE.

Various analytical methods for testing the chemotherapeutic agent 5-fluorouracil (5-FU) in biological fluids of humans and animals have been reported previously. Whereas the administration of 5-FU alter in any way the morphology and function of the salivary glands, and that the presence of chemotherapy in the oral mucosa can lead to some oral complications, this study aimed to determine the presence of 5-FU in chemotherapy treated hamsters saliva by Liquid Chromatography High Performance (HPLC), since this animal model is used in studies of oral mucositis and gland hypofunction induced by 5-FU. Twelve animals were divided into 4 groups: intraperitoneally pilocarpine (CP), pilocarpine + isoproterenol (CPI) and the chemotherapy of the vehicle, and the QP and QPI groups where the animals were, respectively, the same secretagogues listed above and 5-FU chemotherapy. After administration secretagogue, the saliva from all animals was collected for a period of 60 min. Then the saliva was frozen at -80 ºC for subsequent determination of chemotherapy by HPLC. After analysis of chromatograms, and based on the results obtained, it was possible to identify the presence of 5-FU in Hamsters saliva samples that received intraperitoneal chemotherapy via the technique of HPLC.

Antitumor compounds from Streptomyces sp. KML-2, isolated from Khewra salt mines, Pakistan

BACKGROUND: Actinomycetes are gram positive bacteria with high G + C content in their DNA and are capable of producing variety of secondary metabolites. Many of these metabolites possess different biological activities and have the potential to be developed as therapeutic agents. The aim of the present study was to screen actinomycetes inhabiting halophilic environment such as Khewra salt mines present in Pakistan for cytotoxic and antitumor compounds. RESULTS: An actiomycetes strain designated as Streptomyces sp. KML-2 was isolated from a saline soil of Khewra salt mines, Pakistan. The strain Streptomyces sp. KML-2 showed 84 % cytotoxic activity against larvae of Artemiasalina. In the screening phase, the strain exhibited significant antitumor activity with IC50 values of 12, 48 and 56 µg/ml against Hela, MDBK and Vero cell lines, respectively. After that extract from 20 l fermentation was used to purify secondary metabolites by several chromatographic techniques. Structure elucidation of isolated compounds revealed that it is highly stable producer of Chromomycin SA (1) and 1-(1H-indol-3-yl)-propane-1,2,3-triol (2). Both of the isolated compounds showed significant antitumor activity against Hela and MCF-7 cancer cell lines (IC50 values 8.9 and 7.8 µg/ml against Hela; 12.6 and 0.97 µg/ml against MCF-7, respectively). The 16S rRNA gene sequence (1437 bp) of the strain confirm its identity (99 %) with Streptomyces griseus. CONCLUSIONS: From this research work we were successful in isolating two potent antitumor compounds, Chromomycin SA and 1-(1H-indol-3-yl)-propane-1,2,3-triol from Streptomyces KML-2 strain, isolated from Khewra salt mine. As such this is the second report which confirms that S. griseus can produce Chromomycin SA without introducing any mutagenesis in its biosynthesizing gene cluster and isolated indole derivative is being reported first time from any member of actinomycetes group with having novel antitumor activity against Hela and MCF-7 cells Nucleotide sequences: Nucleotide sequence data reported are available in the GenBank database under the accession number: GenBank KJ009562.

Cuantificación de cistina en orina espontánea para la detección de cistinuria / Quantitation of urinary cystine for the detection of cystinuria / Determinagáo quantitativa de cistina urinaria para a detecgáo de cistinuria

La cistinuria es un error innato del metabolismo ocasionado por un defecto en el transporte renal de arginina, ornitina, lisina y cistina. La acumulación de este último aminoácido de baja solubilidad ocasiona episodios de urolitiasis característicos de la enfermedad. En el presente estudio se estandarizó un método espectrofotométrico confiable y de fácil ejecución para la determinación cuantitativa de cistina en orina espontánea. Se realizó el análisis en 184 muestras, correspondientes a 104 controles y 80 pacientes con urolitiasis. Con el objeto de validar el método y posteriormente establecer un rango de excreción normal en la población colombiana se evaluaron los siguientes parámetros: exactitud, precisión, linealidad y límite de detección. La técnica mostró coeficientes de variación intra e inter ensayos inferiores al 10% y una excelente linealidad, con un coeficiente r² entre concentraciones conocidas de cistina y absorbancia generada por el método de 0,998. Usando esta técnica se encontró un valor normal de excreción de 1,35 a 110,11 mg cistina/g creatinina. En cinco pacientes, de los 80 con nefrolitiasis, se hallaron valores elevados de cistina, compatibles con cistinuria. El método utilizado puede implementarse en cualquier laboratorio clínico para confirmar el diagnóstico de cistinuria e iniciar un tratamiento oportuno.

Cystinuria is an inborn error of metabolism, caused by a defect in renal tubular transport of the following aminoacids: arginine, ornithine, lysine and cystine. Accumulation of the latter poorly soluble aminoacid leads to the development of kidney stones, characteristic of the disease. In this study, an easy and dependable spectrophotometric method for the quantitative determination of urinary cystine was standardized. The analysis was performed on 184 samples from 104 controls and 80 patients with kidney stones. In order to validate the method and later establish a range of normal urinary cystine excretion in the Colombian population, the following parameters were evaluated: Accuracy, precision, linearity and lower limit of detection. The technique showed intra and intei assay coefficients of variation below 10%, and excellent linearity, with an R square (r²) coefficient between known cystine concentrations and absorbance generated by the method at 0.998. Using this technique, a normal urinary cystine excretion range of 1.35-110.11 mg cystine/g creatinine was found. Among the 80 patients with kidney stones, elevated urinary cystine levels were found in 5 of them, compatible with the presence of cystinuria. This method can be implemented in any clinical laboratory to confirm the diagnosis of cystinuria and provide opportune treatment.

A cistinúria é um erro inato do metabolismo, causado por um defeito no transporte tubular renal de ar-ginina, ornitina, lisina e cistina. A acumulagáo deste último aminoácido, pouco solúvel, provoca episodios de urolitíase, característicos da doenga. No presente estudo, foi padronizado um método espectrofotomé-trico confiável e de fácil execugáo para a determinagáo quantitativa de cistina em urina espontánea. A análise foi realizada em 184 amostras de 104 controles e 80 pacientes com urolitíase. A fim de validar o método e, posteriormente, estabelecer um intervalo de excregao normal na populagao colombiana, foram avaliados os seguintes parámetros: exatidáo, precisáo, linearidade e limite inferior de detecgáo. O método mostrou coeficientes de variagáo intra e inter ensaios inferiores a 10%, e excelente linearidade, com um coeficiente R quadrado (r²) entre concentragoes conhecidas de cistina e absorváncia gerada pelo método de 0,998. Com esta técnica, foi encontrado um valor normal de excregáo de 1,35-110,11 mg cistina/g de creatinina. Entre os 80 pacientes com urolitíase, foram encontrados níveis elevados de cistina em cinco deles, compatíveis com a presenga de cistinúria. Este método pode ser implementado em qualquer laboratorio clínico para confirmar o diagnóstico de cistinúria e proporcionar um tratamento oportuno.

In vivo evaluation of homeostatic effects of Echis carinatus snake venom in Iran

The venom of the family Viperidae, including the saw-scaled viper, is rich in serine proteinases and metalloproteinases, which affect the nervous system, complementary system, blood coagulation, platelet aggregation and blood pressure. One of the most prominent effects of the snake venom of Echis carinatus (Ec) is its coagulation activity, used for killing prey. Materials and methods Subfractions F1A and F1B were isolated from Ec crude venom by a combination of gel chromatography (Sephadex G-75) and ion exchange chromatography on a DEAE-Sepharose (DE-52). These subfractions were then intravenously (IV) injected into NIH male mice. Blood samples were taken before and after the administration of these subfractions. Times for prothrombin, partial thromboplastin and fibrinogen were recorded. Results and conclusions Comparison of the prothrombin time before and after F1A and F1B administrations showed that time for blood coagulation after injection is shorter than that of normal blood coagulation and also reduced coagulation time after Ec crude venom injection. This difference in coagulation time shows the intense coagulation activity of these subfractions that significantly increase the coagulation cascade rate and Causes to quick blood coagulation. The LD50 of the Ec crude venom was also determined to be 11.1 μg/mouse. Different crude venom doses were prepared with physiological serum and injected into four mice. Comparison of the prothrombin times after injection of subfractions F1A and F1B showed that the rate of mouse blood coagulation increases considerably. Comparing the partial thromboplastin times after injecting these subfractions with this normal test time showed that the activity rate of intrinsic blood coagulation system rose sharply in mice. Finally, by comparing the fibrinogen time after subfraction injections and normal test time, we can.

Glucose(xylose) isomerase production by Streptomyces sp. CH7 grown on agricultural residues

Streptomyces sp. CH7 was found to efficiently produce glucose(xylose) isomerase when grown on either xylan or agricultural residues. This strain produced a glucose(xylose) isomerase activity of roughly 1.8 U/mg of protein when it was grown in medium containing 1% xylose as a carbon source. Maximal enzymatic activities of about 5 and 3 U/mg were obtained when 1% xylan and 2.5% corn husks were used, respectively. The enzyme was purified from a mycelial extract to 16-fold purity with only two consecutive column chromatography steps using Macro-prep DEAE and Sephacryl-300, respectively. The approximate molecular weight of the purified enzyme is 170 kDa, and it has four identical subunits of 43.6 kDa as estimated by SDS-PAGE. Its Km values for glucose and xylose were found to be 258.96 and 82.77 mM, respectively, and its Vmax values are 32.42 and 63.64 μM/min/mg, respectively. The purified enzyme is optimally active at 85ºC and pH 7.0. It is stable at pH 5.5-8.5 and at temperatures up to 60ºC after 30 min. These findings indicate that glucose(xylose) isomerase from Streptomyces sp. CH7 has the potential for industrial applications, especially for high-fructose syrup production and bioethanol fermentation from hemicellulosic hydrolysates by Saccharomyces cerevisiae.

Dengue epidemiology in Thanjavur and Trichy district, Tamilnadu - Jan 2011-Dec 2011.

Background: Dengue infection is emerging as a serious public health problem in Tamil Nadu. An enhanced surveillance system can generate information on the epidemiology of the disease, which is essential for planning and development of relevant control/preventive measures against Dengue. Materials and Methods: A prospective descriptive study was undertaken between January 2011 to December 2011, by testing suspected Dengue patients attending Thanjavur Medical College and Trichy Hospital (TMCH, a major Government referral hospital in Thanjavur District, Tamil Nadu, India) to define the magnitude of Dengue burden, the natural history of this disease in terms of clinical presentation and outcome of the infections in hospitalized Dengue patients. The sera collected from suspected patients were analyzed for Dengue specific IgM and IgG antibodies by IgM antibody capture enzyme linked immunosorbent assay (ELISA) using NIV kit and IgGPanBio Duo Rapid Immunochromatographic Card Test (Brisbane, Australia). The clinical case definition by World Health Organization was adopted to categorize the Dengue cases. Results: The total number of samples screened during the period was 200, out of which 79 (39.5%) were positive for IgM and IgG antibodies and 65 (32%) for IgM antibodies only. By clinical evaluation, Dengue fever was diagnosed in 43 patients, 18 had hemorrhagic manifestations and four patients had progressed to DSS. Though (DSS + DHF) was present in 22 patients, all of them recovered well. Conclusion: In developing countries like India, building of laboratory with advanced capacity for diagnosis and combat-mode ready preparedness for the management of Dengue cases in emergency situation may reduce Dengue-related mortality.