Development and clinical application of hydrogel formulations containing papain and urea for wound healing

Abstract Hydrogels are used for wound treatment, as they may contain one or more active components and protect the wound bed. Papain is one of the active substances that have been used with this purpose, alongside urea. In this paper, carboxypolymethylene hydrogels containing papain (2% and 10% concentrations) and urea (5% concentration) were produced. Physical-chemical stability was performed at 0, 7, 15 and 30 days at 2-8ºC, 25ºC and 40ºC, as well as the rheological aspects and proteolytic activity of papain by gel electrophoresis. Clinical efficacy of the formulations in patients with lower limb ulcers was also evaluated in a prospective, single-center, randomized, double-blind and comparative clinical trial. The results showed 7-day stability for the formulations under 25ºC, in addition to approximately 100% and 15% of protein activity for 10% and 2% papain hydrogel, respectively. The rheological profile was non-Newtonian for the 10% papain hydrogel tested. There were no significant differences regarding the mean time for healing of the lesions, although 10% papain presented a better approach to be used in all types of tissue present in the wound bed.

Caracterização e avaliação da capacidade protetora dos peptí­deos imunogênicos de Sporothrix brasiliensis / Characterization and evaluation of the protective capacity of the immunogenic peptides of Sporothrix brasiliensis

A esporotricose é uma doença crônica que envolve o tecido subcutâneo afetando seres humanos e animais, causada pelo fungo termodimórfico Sporothrix spp.. A esporotricose é endêmica na América latina, principalmente no Brasil que teve o maior surto zoonótico já registrado, ocorrendo na cidade do Rio de Janeiro. A espécie Sporothrix brasiliensis é a mais diagnosticada no surto e a mais virulenta entre as especies de Sporothrix spp., causando formas mais graves da doença. A esporotricose em gatos é endêmica, fatal e um dos principais fatores pelo alto número de casos no Rio de Janeiro. O tratamento é longo e não vem sendo o suficiente para conter o número de casos da doença. Uma vacina contra a esporotricose poderia mudar esse paradigma no Brasil. O presente trabalho obteve o proteoma da cepa S. brasiliensis 5110 por meio de uma eletroforese 2D, e caracterizou e identificou as possíveis proteínas imunogênicas do fungo por espectrometria de massa. Por meio de programas de predição, foi avaliado e sintetizado 7 sequências de aminoácidos,das proteínas identificadas com maiores chances de se acoplar a molécula MHC de classe II. Apenas 3 foram capazes de induzir proliferação in vitro, os peptídeos ZR3, ZR4 e ZR8, que foram utilizados como vacina na esporotricose subcutânea e avaliados sua eficácia por meio da carga fúngica, diâmetro das lesões, perfil celular e níveis de citocinas. Neste trabalho concluímos que o peptídeo ZR8 foi o melhor candidato à vacina na esporotricose, pois foi capaz de diminuir o diâmetro das lesões, aumentar os níveis de citocinas protetoras (IFN-γ, IL-17A e IL-1ß) e aumentar o número de células TCD4+ e CD3-/CD19+, sendo assim induzindo uma resposta imunológica protetora na esporotricose subcutânea

Sporotrichosis is a chronic disease, which involves the subcutaneous tissue affecting humans and animals caused by the thermodymorphic fungus Sporothrix spp. Sporotrichosis is endemic in Latin America, mainly in Brazil that had the largest zoonotic outbreak ever recorded, occurring in the city of Rio de Janeiro. The Sporothrix brasiliensis is the species more diagnosed in the outbreak and most virulent, causing severe forms of the disease. Sporotrichosis in cats is endemic, fatal and the main factors due to the high number of cases of the disease in Rio de Janeiro. The treatment is long, and has not been enough to contain the number of cases of sporotrichosis. A vaccine against sporotrichosis could change this paradigm in Brazil. The present work obtained the proteome of S. brasiliensis 5110 strain by 2D electrophoresis, and characterized and identified possible immunogenic proteins by mass spectrometry. By prediction programs were evaluated and synthesized 7 peptide sequence from antigenic proteins that have the highest chances of coupling to the MHC class II molecule. From these 7 peptides only 3 were able to induce proliferation in vitro, called ZR3, ZR4 and ZR8 peptides, that were used as a vaccine in subcutaneous sporotrichosis and evaluated their efficacy through fungal load, lesion diameter, cell profile and cytokine levels. We conclude that ZR8 peptide was the best candidate for sporotrichosis vaccine, since it was able to decrease the lesion diameter, increase the levels of protective cytokines (IFN-γ, IL-17A and IL-1ß) and increase the number of CD4+ T cells and CD3-/CD19+ inducing a protective immune response in subcutaneous sporotrichosis

Electroforesis de campo eléctrico pulsado en la tipificación molecular de cepas aisladas de un brote de Salmonelosis ocasionado por consumo de queso contaminado con Salmonella Javiana / Pulse Field Gel Electrophoresis in molecular typification of strains isolated from a Salmonellosis outbreak caused by consumption of Salmonella Javiana contaminated cheese

El objetivo de este trabajo fue confirmar por Electroforesis de Campo Electrico Pulsado (PFGE) que la Salmonella aislada del alimento implicado en el brote (queso blanco) fue la responsable del evento.La muestra de queso blanco presento elevado recuento de coliformes, E. coli y S. aureus,ademas, presencia de Salmo nella spp., lo que indico condiciones sanitarias inadecuadas y posible contaminacion de origen fecal. Para la confirmacion de las cepas sospechosas de Sal monella spp, aisladas de los pacientes y del alimento, se utilizaron tecnicas bioquimicas convencionales, la serotipificacion se realizo siguiendo el esquema de White-kauffmann-LeMinor y la sensibilidad a los antimicrobianos (Ampicilina, Trimetroprim-Sulfametoxazole, Ciprofloxacina, Amoxicilina, Ac.Clavulanico, Cloranfenicol, Ceftriaxone y Tetraciclina) por la tecnica kirby-Bauer. Las cepas bacterianas de Salmonella spp aisladas fueron identificadas como Salmonella Javiana [1,9,12:l, z28:1,5] y resultaron sensibles a todos los antibioticos probados.La Tipificacion Molecular de las cepas, se realizo por PFGE, se gun protocolo estandarizado de PulseNet para Salmonella y el analisis de los patrones se estudio utilizando el programa BioNumerics, version 4.0, lo cual mostro que los aisla dos de Salmonella Javiana procedentes tanto de pacientes como del alimento tenian identico patron de restriccion, en tamano y numero de fragmentos. La ocurrencia de un patron unico de PFGE (Coeficiente de similitud 100%) entre los aislados permitio demostrar que el queso contaminado con Salmonella Javiana fue el responsable del brote familiar.

The aim of this work was confirmation through Pulse Field Gel Electrophoresis (PFGE) that Salmonella isolated from the food implicated in the outbreak (white cheese) was responsible for the event. The white cheese sample showed a high count of Coli orms,f E. coli and S. aureus. Moreover, Salmonella was pre ent,s which indicated inadequate sanitary conditions and possible fecal contamination. The suspected Salmonella strains isolated from patients and the food sample, were confirmed using conventional biochemical techniques, serotyping according to the White-kauffmann-Le Minor scheme and antibiotics sensibility (Am picillin, Trimetroprim-Sulfametoxazole, Ciprofloxacin, Amoxicillin, Clavulanic Acid, Cloranfenicol, Ceftriaxone and tetraciclin) following kirby-Bauer’s technique. The Salmonella strains were identified as Salmonella Javiana [1,9,12:I, z28:1,5]and were sensitive to all the antibiotics tested. The molecular typing of the strains was performed using PFGE, according to the PulseNet standardized protocol for Salmonella. The pattern analysis was studied using Bionunella merics program, version 4.0, which showed that the Sal monella Javiana isolates from patients as the food sample had an identical restriction pattern in size and fragments number. The incidence of a unique pattern of PFGE (similarity coefficient 100%) between isolates demonstrated that the cheese contaminated with Salmonella Javiana was responsible for the family outbreak.

Electroforesis capilar / Capillary electrophoresis

En el presente trabajo se detallan los principios físico-químicos básicos que rigen la separación de sustancias en la electroforesis capilar. Se destaca el aporte que ha significado el desarrollo de los capilares de 20 a 50 µm de diámetro, de los detectores con arreglos de diodos y la operativa computacional para la interpretación gráfica de los datos obtenidos. Se detallan los principios que rigen la electroforesis libre, zonal, isotacoferesis, isoelectroenfoque y cromatografía micelar con surfactantes y ciclodextrinas, y distintos ejemplos sobre sus aplicaciones

Psammomys obesus cretzschmar, 1828 and zoonotic cutaneous leishmaniasis in Sinai peninsula, Egypt

In the Middle East, the fat sand rat Psammomys obesus is the most important reservoir host of zoonotic cutaneous leishmaniasis [ZCL]. It is the most incriminated host in outbreaks. Two P. obesus caught in Wadi El-Gedeiret [El-Arish, North Sinai] were found naturally infected with Leishmania major as indicated by enzyme electrophoresis. In Egypt, the already known reservoir hosts are Gerbillus pyramidum I, Geoffroy St. Hilaire, 1825, Meriones crassus Sundevall, 1984 and Meriones sacramnti Thomas, 1922. The role of P. obesus as a host was discussed

Purification and characterization of a specific fasciola antigen

Specific fasciola antigen was prepared from homogenates of Fasciola hepatica adult worms. The homogenate was ultracentrifuged and the supernatant containing crude fasciola antigen was then passed over a cyanogen bromide activated sepharose 4B column coupled with antiserum against Schistosoma mansoni adult worm surface antigen. The specific, schistosoma-free fasciola antigen was tested for its specificity by immunodiffusion. Characterization of the specific fasciola antigen was done by gradient polyacrylamide gel electrophoresis and immunoblotting technique. The electrophoresis migration pattern of specific fasciola antigen, stained with Coomassie blue, showed seven bands in the 12-54 kDa regions. Using the immunoblotting technique, a batch of positive fascioliasis sera recognized two specific bands at the 33 and 54 kDa regions

Determination of G /A +G ratio in beta thalassemia, an indicator of diseases severity

The globin chains were separated by electrophoresis on acrylamide-bis acrylamide gels containing Triton X-100, urea, 2 mercaptoethanol, and acetic acid. The G gamma/ A gamma+ G gamma ratio was determined in 25 cord blood samples and in 46 beta-thalassemic homozygotes among Iranian children. In 25 cord blood samples the mean value for G gamma / ? gamma + G gamma was 0.66. All beta-thalassemia homozygotes had an elevated HbF from 5-80% and the mean value for G gamma / G gamma + A gamma was 0.64 and their G gamma chain level in the peripheral blood was found to be intermediate between the embryonic and adult values. We conclude that HbF and G gamma chain level as well as the severity of the disease is due to Beta-thalassemia variants, but there is a poor correlation between HbF level and G gamma /G gamma + ? gamma ratio, both in cord blood samples and Beta-thalassemia samples

Electrophoretic fractions of the serum proteins in malaria

Forty five cases of malaria, thirty cases of non-malarial pyrexia and twenty five normal controls were studied to find out quantitative differences of total serum proteins and its major electrophoretic fractions in different groups and to determine the level of humoral immunity. The results showed significant decrease in total serum proteins and increase in gamma globulins, thus, supporting the general belief that there is a correlation between immunity and malaria

three year study of the electrophoretic patterns of serum proteins with special reference to various globulinopathies in Saudis

Human sera comprising 545 specimens spread over a period of three years were analyzed electrophoretically for proteins. Only 159 samples [29%] exhibited abnormal serum protein patterns. They were in the order of inflammatory disorders [acute/chronic], 60 [37.7%], Polyclonal gammopathies,48 [30.1%], Cirrhosis, 22 [13.8%], Monoclonal gammopathies, 22 [13.8%], Agammaglobulinemia, 3 [1.9%], Nephrosis, 2 [1.2%], Protein-losing enteropathy, 1 [0.6%], Low-alpha 1 -antitrypsin, 1 [0.6%], Inflammatory diseases of various nature, therefore, constitute the major serum protein abnormalities in Riyadh. Moreover, their occurrence varies with age, sex and climatic conditions as elsewhere in the world

Secretory immunoglobulin in lactating women

Serum and milk total proteins, its electrophretic fractions and immunoglobulins [IgA-IgM] were determined in eleven healthy pregnant women in their third trimester and up to the end of the third month postpartum. Values of serum total proteins, albumin and gamma-globulins show a significant fall in late pregnancy returned to normal levels after pureperium. Total globulins and alpha 2-globulin and beta-globulin significantly increase in late pregnancy reaching the maximum by 3-4 days postpartum, returned to nearly normal by the end of the third month postpartum; As regards to immunoglobulins, IgA values showed insignificant increase while IgM significantly increase through the follow up period. As regards to colostrum and milk the highest concentration of total proteins, globulins and immunoglobulins were in colostrum, these concentrations declined during the first 3 months of lactation. These changes show that Human colostrum and milk are rich sources of immunoglobulins particularly of serum IgA

Un prototipo para la electroforesis de campo pulsante. Estandarización de las condiciones para la resolución de grandes moléculas de ADN / A prototype for the pulse field electrophoresis: condition for the resolution of large DNA molecules

Se desarrolló un prototipo para la electroforesis de campo pulsante en gel de agarosa, que permite separar moléculas de ADN de hasta 2 000 kb. El equipo se fundamenta en la propiedad que tienen las moléculas grandes de ADN, de fraccionarse en geles de agarosa si son sometidas alternadamente a dos campos eléctricos no homogéneos de orientación perpendicular. En los experimentos, los tiempos de pulso fueron 60s, y se estudió la influencia de la fuerza iónica y el tiempo de corrida sobre la resolución de las bandas. El método fue utilizado para obtener el cariotipo electroforético de dos cepas de Saccharomyces cerevisiae. El trabajo describe las distintas partes del equipo, el protocolo electroforético, y la caracterización parcial de algunas bandas mediante hibridización ADN-ADN con sondas específicas para diferentes cromosomas