Pythiosis cutánea en equinos tratados con acetonida de triamcinolona. Parte 2. Descripción histológica e histoquímica

Objective. The study aimed to evaluate the histomorphometry tissue recovery process of the skin granuloma of skin pythiosis in horses treated with triamcinolone acetonide. Materials and methods. We conducted a descriptive study, not probabilistic in convenience animals with cutaneous pythiosis. 24 horses were used with cutaneous pythiosis, a group of 12 animals was administered 50 mg of intramuscular injection of triamcinolone acetonide (TG) and the other group was not applied any treatment (CG). Are tissue biopsies performed for histological and histochemical evaluation and stained with hematoxylin eosin (HE), Gomori trichrome (GT), picrosirius red polarization (PR/P), Grocott methenamine silver (GMS) and periodic acid-Schiff (PAS). Results. It is noted that in TG inflammation was gradually decreasing, as evidenced in decreased fibrin layer leukocyte, PMN and phenomena Splendore Hoepli, also in increased angiogenesis, epiteliogénesis, and increasing the overall amount of fibroblasts and collagen fibers, anyway in the progressive replacement of collagen type III to type I collagen at the end of the process, and that the presence of intralesional pseudo-hyphae of Pythium insidiosum reduces it to the second week. Neither of the animals in the CG showed improvement in histological and histochemical characteristics of pythiosis and maintained equal to the first day throughout the study. Conclusions. The use of triamcinolone acetonide is a good therapeutic alternative for the treatment of granulomatous pythiosis wounds in horses with 100% clinical recovery and demonstrated with histological and histochemical findings.Objetivo. El objetivo del estudio fue realizar una descripción histológica e histoquímica del proceso de cicatrización de la pythiosis cutánea en equinos tratados con acetonida de triamcinolona. Materiales y métodos. Se Realizó un estudio de tipo descriptivo, no probabilístico en animales de conveniencia con pythiosis cutánea. Fueron utilizados 24 equinos con pythiosis, siendo aplicado 50 mg de acetonida de triamcinolona vía intramuscular a un grupo de 12 animales (GT) y en otro grupo no fue aplicado tratamiento (GC). Se realizaron biopsias de tejidos para evaluación histológica e histoquímica, en las coloraciones de hematoxilina eosina (H&E), tricrómico de Gómori (TG), picrosirius red/polarization (PR/P) y plata metanamina de Grocott (GMS). Resultados. Se observó que en el GT el proceso inflamatorio fue disminuyendo progresivamente, evidenciado en la disminución de la capa fibrino leucocitaria, PMN y de los fenómenos de Splendore Hoepli, así como mayor angiogénesis, epiteliogénesis y aumento de la cantidad de fibroblastos y fibras colágenas generales, así mismo en el cambio progresivo de colágeno tipo III a colágeno tipo I al final del proceso, además de que la presencia de hifas intralesionales de Pythium insidiosum disminuyó a la segunda semana. Ninguno de los animales del GC presentó mejoría en las características histológicas e histoquímicas de la pythiosis y se mantuvieron iguales al primer día durante todo el estudio. Conclusiones. La acetonida de triamcinolona es una buena alternativa terapéutica en el tratamiento de las heridas granulomatosas por pythiosis en equinos con 100% de recuperación clínica y demostrada con las constataciones histológicas e histoquímicas

Pythiosis cutánea en equinos tratados con acetonida de triamcinolona. Parte 3. Análisis histomorfométrico

Objective. The objective of the study was to analyze Histomorphometrically of the healing process with cutaneous pythiosis in horses treated with triamcinolone acetonide. Materials and methods. 24 horses with pythiosis were used, to a group 50 mg of intramuscular triamcinolone acetonide (GT) was applied, while the other group was not applied treatment (GC). They were collected tissue biopsies, processed, sliced and stained with hematoxylin and eosin (H & E), Gomori trichrome (TG), picrosirius red / polarization (PR / P) and Grocott methenamine silver (GMS). Photomicrographs were selected and 10 histological changes, analyzed with BioEstat 5.0 software, obtaining quantities of tissue cells such as eosinophils, neutrophils, macrophages, fibroblasts and collagen through planimetric evaluation point count. Results. In GSM staining was observed decrease in the presence of intralesional hyphae of P. insidiosum to 16 days (p<0.05). Staining H&E, we observed a decrease of the inflammatory process, shown in eosinophils (p=0.0001), neutrophils (p=0.0001), and macrophage (p=0.00001). In the staining of GT and PR/P increase the amount of fibroblasts and collagen fibers were observed, also the gradual exchange of type III collagen to type I, increased fibroblast show significant (p=0.0001) from day 16 until day 40, the expression of collagen was significant (p=0.0001) from day 16 until the end of the study. It was statistically significant correlation between neutrophils and macrophages (p=0.00018), collagen and eosinophil (p=0.03) and fibroblasts and collagen (p=0.02). The animals in the CG do not present histomorphometric improvement during the study. Conclusions. We conclude that the cell produces triamcinolone acetonide and histomorphometric tecidual recovery in horses with pythiosis.Objetivo. El objetivo del estudio fue analizar histomorfométricamente el proceso de cicatrización en equinos con pythiosis tratados con acetonida de triamcinolona. Materiales y métodos. Se utilizaron 24 equinos con pythiosis, a un grupo se aplicó 50 mg de acetonida de triamcinolona intramuscular (GT), mientras que al otro grupo no fue aplicado tratamiento (GC). Fueron colectadas biopsias de tejido, procesadas, laminadas y coloreadas con hematoxilina eosina (H&E), tricrómico de Gómori (TG), picrosirius red/polarization (PR/P) y plata metanamina de Grocott (GMS). Fueron seleccionados y fotomicrografiados 10 campos histológicos, analizados con el software BioEstat 5.0, obteniéndose las cantidades tisulares de células como eosinófilos, neutrófilos, macrófagos, fibroblastos y colágeno a través de evaluación planimétrica por conteo de puntos. Resultados. En coloración de GSM, se observó disminución de hifas intralesionales del Pythium a los 16 días (p<0.05). En coloración de H&E, hubo disminución del proceso inflamatorio, evidenciado en eosinófilos (p=0.0001), neutrófilos (p=0,0001) y macrófagos (p=0.00001). En coloraciones de TG y PR/P, hubo aumento de fibroblastos, fibras colágenas y cambio progresivo de colágeno tipo III a tipo I, el aumento de fibroblastos fue significativo (p=0.0001) del día 16 al 40, la expresión de colágeno fue altamente significativa (p=0.0001) del día 16 hasta el final del estudio. Hubo correlación significativa entre neutrófilos y macrófagos (p=0.00018), colágeno y eosinófilos (p=0.03) y colágeno y fibroblastos (p=0.02). Los animales del GC no presentaron mejoría histomorfométrica durante el estudio. Conclusiones. Es posible concluir que la acetonida de triamcinolona produce recuperación histomorfométrica celular y tecidual en los equinos con pythiosis

Pythiosis cutánea en equinos tratados con acetonida de triamcinolona. Parte 1. Caracterización clínica

RESUMEN Objetivo. El objetivo del estudio fue caracterizar clínicamente el proceso de cicatrización de la pythiosis cutánea en equinos tratados con acetonida de triamcinolona. Materiales y métodos. Se realizó un estudio descriptivo, no probabilístico, de conveniencia. Fueron utilizados 24 equinos con pythiosis cutánea. A un grupo le fue aplicado 50 mg de acetonida de triamcinolona vía intramuscular (GT) y al otro grupo no fue aplicado tratamiento (GC). Se realizó caracterización clínica y macroscópica de la lesión cada 4 días hasta la recuperación completa del granuloma en el GT. Siendo verificada la disminución del exudado fibrinosanguinolento, prurito, presencia de trayectos fistulosos y salida de kunkers, también, fueron realizadas proyecciones radiológicas para evaluar el grado de compromiso óseo y fibrosis tecidual. Resultados. Macroscópicamente, los animales del GT presentaron diminución progresiva en las características clínicas de la enfermedad a los 16±1,4 días post aplicación del tratamiento. La contracción de la herida fue del 100% a los 60±3.4 días post aplicación del tratamiento en todos los granulomas del GT, siendo altamente significativa (p<0.0001). Radiológicamente, se encontró invasión a los tejidos subyacentes comprometiendo estructuras óseas, caracterizándose por fibrosis tisular, exostosis moderada, osteólisis y osteomielitis. Los animales del GC no presentaron mejoría en las características clínicas y macroscópicas de la pythiosis y se mantuvieron iguales durante todo el estudio. Conclusiones. La acetonida de triamcinolona es una buena alternativa terapéutica en el tratamiento de las heridas granulomatosas de pythiosis en equinos con 100% de recuperación clínica

Strong-LAMP Assay Based on a Strongyloides spp.-Derived Partial Sequence in the 18S rRNA as Potential Biomarker for Strongyloidiasis Diagnosis in Human Urine Samples

Human strongyloidiasis a soil-transmitted infection caused by Strongyloides stercoralis is one of the most neglected amongst the so-called Neglected Tropical Diseases (NTDs). S. stercoralis is a nematode, which is distributed worldwide; it has been estimated that it could affect millions of people, mainly in tropical and subtropical endemic regions. The difficulties of diagnosis lead to infection rates being underreported. Asymptomatic patients have chronic infections that can lead to severe hyperinfection syndrome or disseminated strongyloidiasis in immunocompromised patients. Strongyloidiasis can easily be misdiagnosed because conventional faecal-based techniques lack of sensitivity for the morphological identification of infective larvae in faeces. None of the currently used molecular methods have used urine samples as an alternative to faecal samples for diagnosing strongyloidiasis. This study was thus aimed at comparing, for the first time, the use of a new loop-mediated isothermal amplification (LAMP) molecular assay (Strong-LAMP) to traditional methods on patients' urine samples. Twenty-four urine samples were taken from patients included in a study involving two Spanish hospitals for strongyloidiasis screening using parasitological and serological tests. Strongyloides larvae were found in 11 patients' faecal samples, thereby ascertaining that they had the disease. Other patients had high antibody titres but no larvae were found in their faeces. All urine samples were analysed by PCR and Strong-LAMP assay. No amplification occurred when using PCR. Strong-LAMP led to detecting S. stercoralis DNA in urine samples from patients having previously confirmed strongyloidiasis by parasitological tests and/or a suspicion of being infected by serological ones. The Strong-LAMP assay is a useful molecular tool for research regarding strongyloidiasis in human urine samples. After further validation, the Strong-LAMP assay could also be used for complementary and effective diagnosis of strongyloidiasis in a clinical setting

Healing Effects of Formulations with Extract of Sonchus oleraceus

Introduction: Scar is defined as a process of tissue regeneration characterized by fibroblasts and a disorganized extracellular matrix, formed mainly by collagen. Thus it is understood that regeneration capacity is a phenomenon that occurs in order to repair an injured tissue. However, science has been looking for alternatives to assist in the repair process, and thus phytotherapy has been used as a therapeutic resource in medicine, and scientifically proven its efficiency. The plant Sonchus oleraceus is known, among other names, as “serralha” or sowthistle. It is a cosmopolitan species, found in several agricultural regions infesting crops. In Brazil, it is more frequent to grow in the winter and its seeds can be kept viable for eight years in the soil. This plant can be used as a popular drug for diuretic proposals, in cases of anemia, liver diseases and in cases of treatment of vitiligo in the eastern region of the state of Minas Gerais, Brazil. The objective of this research was to evaluate two different products (ointment and balsam) composed by S. oleraceus in the healing of surgical wounds in rats (Rattus novergicus).Materials, Methods & Results: Sonchus oleraceus specimens were obtained at the beginning of May, during the autumnwinter season, in which there is already a flowering of this plant. The leaves and stem of S. oleraceus were selected, cleaned and dried under study at 40°C of circulating air for four days for complete dehydration, and then ground in a knife mill. The constituents were prepared in 80% alcohol (v / v) and subjected to ultrasonic extraction for 60 min. The hydroalcoholic extract of S. oleraceus was concentrated on a rotary evaporator until all alcohol evaporated and the aqueous extract was obtained. The extracts were produced at concentrations of 5%, 10% and 15% and sent for manipulation of the pharmaceutical formulations of ointment and salve. Forty Wistar rats were divided into eight groups for evaluation of different products (salve and ointment). The wounds were created on the back of the animals and the products were applied daily; for analysis, clinical evaluations and wound measurement were performed daily. After 15 days, a biopsy was performed for histopathological analysis and quantification of type I and III collagen fibers. Measurement of the wound in rats treated with ointment containing 15% of extract presented better tissue retraction, with complete retraction observed after 10 days of treatment. Type I collagen was predominant in all samples; however, the healing properties of the balsam containing 5% extract and ointment containing 5% and 15% extract were significantly different.Discussion: As a conclusion, Sonchus oleraceus influenced the healing process of the skin. This pharmacological action is probably directly related to the amount of flavonoids present in its composition, that is, the higher the extract concentration, the better its efficacy. However, a better efficacy in the ointment is emphasized by better adhesion to the skin. Studies have shown that the flavonoids present in most plants have relevant healing activity, stimulating the proliferation of fibroblasts and the production of collagen, acting on the inhibition of metalloproteinase, thus increasing the rate of synthesized collagen. Other researchers reported a high concentration of flavonoids (anthocyanins and tannins) that act as mediators of inflammation, stimulating the production of inflammatory cytokines and tumor necrosis factor, observed in this species

Pythiosis cutaneous in horses treated with triamcinolone acetonide. Part 2. Histological and histochemical description

ABSTRACT Objective. The study aimed to evaluate the histomorphometry tissue recovery process of the skin granuloma of skin pythiosis in horses treated with triamcinolone acetonide. Materials and methods. We conducted a descriptive study, not probabilistic in convenience animals with cutaneous pythiosis. 24 horses were used with cutaneous pythiosis, a group of 12 animals was administered 50 mg of intramuscular injection of triamcinolone acetonide (TG) and the other group was not applied any treatment (CG). Are tissue biopsies performed for histological and histochemical evaluation and stained with hematoxylin eosin (HE), Gomori trichrome (GT), picrosirius red polarization (PR/P), Grocott methenamine silver (GMS) and periodic acid-Schiff (PAS). Results. It is noted that in TG inflammation was gradually decreasing, as evidenced in decreased fibrin layer leukocyte, PMN and phenomena Splendore Hoepli, also in increased angiogenesis, epiteliogénesis, and increasing the overall amount of fibroblasts and collagen fibers, anyway in the progressive replacement of collagen type III to type I collagen at the end of the process, and that the presence of intralesional pseudo-hyphae of Pythium insidiosum reduces it to the second week. Neither of the animals in the CG showed improvement in histological and histochemical characteristics of pythiosis and maintained equal to the first day throughout the study. Conclusions. The use of triamcinolone acetonide is a good therapeutic alternative for the treatment of granulomatous pythiosis wounds in horses with 100% clinical recovery and demonstrated with histological and histochemical findings.RESUMEN Objetivo. El objetivo del estudio fue realizar una descripción histológica e histoquímica del proceso de cicatrización de la pythiosis cutánea en equinos tratados con acetonida de triamcinolona. Materiales y métodos. Se Realizó un estudio de tipo descriptivo, no probabilístico en animales de conveniencia con pythiosis cutánea. Fueron utilizados 24 equinos con pythiosis, siendo aplicado 50 mg de acetonida de triamcinolona vía intramuscular a un grupo de 12 animales (GT) y en otro grupo no fue aplicado tratamiento (GC). Se realizaron biopsias de tejidos para evaluación histológica e histoquímica, en las coloraciones de hematoxilina eosina (H&E), tricrómico de Gómori (TG), picrosirius red/polarization (PR/P) y plata metanamina de Grocott (GMS). Resultados. Se observó que en el GT el proceso inflamatorio fue disminuyendo progresivamente, evidenciado en la disminución de la capa fibrino leucocitaria, PMN y de los fenómenos de Splendore Hoepli, así como mayor angiogénesis, epiteliogénesis y aumento de la cantidad de fibroblastos y fibras colágenas generales, así mismo en el cambio progresivo de colágeno tipo III a colágeno tipo I al final del proceso, además de que la presencia de hifas intralesionales de Pythium insidiosum disminuyó a la segunda semana. Ninguno de los animales del GC presentó mejoría en las características histológicas e histoquímicas de la pythiosis y se mantuvieron iguales al primer día durante todo el estudio. Conclusiones. La acetonida de triamcinolona es una buena alternativa terapéutica en el tratamiento de las heridas granulomatosas por pythiosis en equinos con 100% de recuperación clínica y demostrada con las constataciones histológicas e histoquímicas