Immunomodulatory activity of itraconazole in lung

Purpose: To evaluate the in vivo effects of ITC as an immunomodulator on various immunological mediators, including cytokines, chemokines and growth factors in pulmonary homogenates.Methods: Two experimental groups consisting of 25 BALB/c mice each were used: (a) PBS-treated mice and (b) ITC-treated mice (1 mg/mouse/day). The animals were treated daily via gavage for 8 weeks. Five mice from each group were sacrificed at 0, 4, 8, and 12 weeks and cytokine levels assessed in the supernatants of lung macerates using the Bio-Plex system. Five lungs from each experimental group, after 8 weeks of treatment, were used for histopathological analysis.Results: Compared with the control group, ITC-treated mice showed significant changes in the pulmonary levels of 50 % of the molecules evaluated: IL-4, IL-10, IL-12p70, IL-13, TNF-α, eotaxin, MIP- 1α, MIP-2, and LIF were significantly upregulated. In contrast, IL-1β, IL-15, IL-2, IL-1α, and VEGF were downregulated.Conclusion: Although histopathological examination did not show changes in lung cell infiltrates, ITC exerted a marked immunomodulatory effect at the pulmonary level in healthy BALB/c mice.Keywords: Immunomodulation, Itraconazole, Antifungal drug, Histopathological examinatio

Synthesis and in-vitro evaluation of s-allyl cysteine ester-caffeic acid amide hybrids as potential anticancer agents

We have synthesized a series of S-allyl cysteine ester-caffeic acid amide hybrids and evaluated them in order to determine their possible anticancer activity and selectivity in colorectal cancer, which is still one of the leading causes of morbidity and mortality worldwide. All compounds were tested against SW480 human colon adenocarcinoma cells and the non-malignant CHO-K1 cell line. Among the tested compounds, hybrids 6e, 9a, 9b, 9c and 9e exhibited the highest effect on viability (IC50 SW480-48h= 0.18, 0.12, 0.12, 0.11 and 0.12 mM, respectively) and selectivity (SI= 10.3, 1.5, >83.33, >90.91 and >83.33, respectively) in a time- and concentration-dependent manner. Besides, our results were even better as regards lead compounds (S-allyl cysteine and caffeic acid) and the standard drug (5-FU). Additionally, these five compounds induced mitochondrial depolarization that could be related with an apoptotic process. Moreover, hybrids 6e, 9a and 9e induced cell cycle arrest in G2/M phase, and compound 9c in S- phase, which suggests that these hybrid compounds could have also a cytostatic effect in SW480 cell line. The SAR analysis showed that hydroxyl groups increased the activity, besides, there was not a clear relationship between the antitumor properties and the length of the alkyl chain. Since hybrid compounds were much more selective than the conventional drug (5-FU), this make them promising candidates for further studies against colorectal cancer

Synthesis and Antiproliferative Activity of 3 and 7-Styrylcoumarins

A series of styrylcoumarins were obtained via Mizoroki-Heck reactions between 3-bromo-4-methyl7-(octyloxy)-2H-chromen-2-one or 2-oxo-2H-chromen-7-yl trifluoromethanesulfonate and functionalized styrenes. The structures of the products were elucidated by spectroscopic analysis. All compounds were evaluated against SW480 and CHO-K1 cell lines. A number of hybrids showed good antiproliferative activity. Among the tested compounds, hybrids 6e, 10c and 10d, exhibited the highest activity (IC50- SW480/48h = 6,92; 1,01 and 5,33 µM, respectively) and selectivity (IS48h = >400; 67,8 and 7,2, respectively). In addition, these compounds were able to preserve their activities over time. The results achieved by these hybrids were even better than the lead compounds (coumarin and resveratrol) and the standard drug (5-FU). As regards structure-activity relationship it seems that the location of the styryl group on the coumarin structure and the presence of the hydroxyl group on the phenyl ring were determinant for the activity

Estandarización y validación de un método de cromatografía líquida de alta eficiencia con detector de arreglo de diodos (HPLC-DAD) para la determinación de niveles sanguíneos de voriconazol

Introduction. A specialized service for the determination of antifungal blood levels is not available in Colombia, this service is essential for the proper follow-up of the antifungal therapy.Objective. The aim of this study was to standardize and validate a simple, sensitive, and specific protocol, based on High Performance Liquid Chromatography with Diode Array Detector (HPLC-DAD) for the quantification of voriconazole blood levels. Materials and methods. An Agilent HPLC-series-1200 equipment with UV-DAD detector was used. Analytical column-Eclipse XDB-C18 and pre- column Eclipse-XDB-C18, Agilent were also used. We used voriconazole as the primary control, and posaconazole as an internal control. The validation was done following the Food and Drug Administration (FDA) recommendations.Results. Best chromatographic conditions were: Column temperature of 25°C, UV variable wavelength detectors (VWD) detection at 256 nm for voriconazole, and 261 nm for Posaconazole (internal standard), 50 μL of injection volume, flow of volume 0,8mL/min, time of run of 10 min, mobile phase of acetonitrile:water (60:40). Finally, retention times were 3.13 and 5.16 min for the voriconazole and posaconazole, respectively. Range of quantification ranging from 0.125 μg/mL to 16 μg/mL. Conclusion. The selectivity and chromatographic purity of the obtained signal as well as the limits of detection and quantification standardized make this method an excellent tool for therapeutic monitoring of patients treated with voriconazole.Introducción. Hasta la fecha en Colombia no contábamos con un servicio especializado de medición de niveles séricos de antifúngicos, procedimiento esencial para el adecuado manejo del tratamiento de las Infecciones Fúngicas Invasoras (IFI). Objetivo. Estandarizar y validar un protocolo, simple, sensible y específico, basado en Cromatografía Líquida de Alta Eficiencia con detector de arreglo de diodos (HPLC-DAD) para la cuantificación de los niveles séricos de voriconazol. Materiales y métodos. Se usó un equipo HPLC-Agilent, serie-1200, con detector UV-DAD columna analítica Eclipse-XDB-C18 y una pre-columna Eclipse-XDB-C18, ambas de la marca Agilent. Como control primario se utilizó voriconazol, y como control interno posaconazol. La validación se hizo cumpliendo todos los criterios de aceptación de los parámetros recomendados por la Food and Drug Administration (FDA). Resultados. Las mejores condiciones cromatográficas se obtuvieron bajo los siguientes parámetros: temperatura de la columna de 25°C, detección UV-VWD de 261 nm, volumen de inyección de 50 μL, flujo de 0,8mL/min y un tiempo de corrido de 10 min. La fase móvil usada fue acetonitrilo:agua (40:60), obteniendo finalmente unos tiempos de retención de 3,13 y 5,16 min para el voriconazol y posaconazol respectivamente. El rango de cuantificación fue desde 0,125 μg/mL hasta 16 μg/mL. Conclusiones. La selectividad y pureza de la señal cromatográfica obtenida, así como los límites de detección y cuantificación estandarizados, hacen de esta metodología una excelente herramienta para el seguimiento terapéutico de los pacientes bajo tratamiento o profilaxis con voriconazol

Pulmonary Abnormalities in Mice with Paracoccidioidomycosis: A Sequential Study Comparing High Resolution Computed Tomography and Pathologic Findings

Paracoccidioidomycosis (PCM) is a fungal infection caused by the dimorphic fungus Paracoccidioides brasiliensis. It occurs preferentially in rural workers in whom the disease is severe and may cause incapacitating pulmonary sequelae. Assessment of disease progression and treatment outcome normally includes chest x-rays or CT studies. Existing experimental PCM models have focused on several aspects, but none has done a radiologic or image follow-up evaluation of pulmonary lesions considered as the fungus primary target. In this study, the lungs of mice infected with fungal conidia were studied sequentially during the chronic stage of their experimental mycosis by noninvasive high resolution medical computed tomography, and at time of sacrifice, also by histopathology to characterize pulmonary abnormalities. Three basic lung lesion patterns were revealed by both techniques: nodular-diffuse, confluent and pseudo-tumoral which were located mainly around the hilus thus accurately reflecting the situation in human patients. The experimental design of this study decreases the need to sacrifice a large number of animals, and serves to monitor treatment efficacy by means of a more rational approach to the study of human pulmonary diseases. The findings we are reporting open new avenues for experimental research, increase our understanding of the mycosis pathogenesis and consequently have repercussions in patients' care

Structural and Topographic Dynamics of Pulmonary Histopathology and Local Cytokine Profiles in Paracoccidioides brasiliensis Conidia-Infected Mice

Paracoccidioidomycosis (PCM), an endemic fungal infection of pulmonary origin resulting in severe disseminated disease, occurs in rural areas of most South American countries and presents several clinical forms. The infection is acquired by inhalation of specific fungal propagules, called conidia. Considering the difficulties encountered when studying the infection in humans, this work was done in mice infected by inhalation of infective fungal conidia thus mimicking the human natural infection. The lungs of mice were sequentially studied by histopathological and multiplex cytokine methods from 2 h to 16 weeks after infection to verify the course of the disease. The mycosis presented different morphologic aspects during the course of time, affecting several pulmonary compartments. Otherwise and based on the analysis of 30 cytokines, the immune response also showed heterogeneous responses, which were up or down regulated depending on the time of infection. By recognizing the different stages that correspond to the evolution of pulmonary lesions, the severity (benign, chronic or fibrotic) of the disease could be predicted and the probable prognosis of the illness be inferred

Recent Advances in Polymer Nanomaterials for Drug Delivery of Adjuvants in Colorectal Cancer Treatment: A Scientific-Technological Analysis and Review

Colorectal cancer (CRC) is the type with the second highest morbidity. Recently, a great number of bioactive compounds and encapsulation techniques have been developed. Thus, this paper aims to review the drug delivery strategies for chemotherapy adjuvant treatments for CRC, including an initial scientific-technological analysis of the papers and patents related to cancer, CRC, and adjuvant treatments. For 2018, a total of 167,366 cancer-related papers and 306,240 patents were found. Adjuvant treatments represented 39.3% of the total CRC patents, indicating the importance of adjuvants in the prognosis of patients. Chemotherapy adjuvants can be divided into two groups, natural and synthetic (5-fluorouracil and derivatives). Both groups can be encapsulated using polymers. Polymer-based drug delivery systems can be classified according to polymer nature. From those, anionic polymers have garnered the most attention, because they are pH responsive. The use of polymers tailors the desorption profile, improving drug bioavailability and enhancing the local treatment of CRC via oral administration. Finally, it can be concluded that antioxidants are emerging compounds that can complement today’s chemotherapy treatments. In the long term, encapsulated antioxidants will replace synthetic drugs and will play an important role in curing CRC

Validation and clinical application of a molecular method for the identification of Cryptococcus neoformans/Cryptococcus gattii complex DNA in human clinical specimens

ABSTRACT The diagnosis of cryptococcosis is usually performed based on cultures of tissue or body fluids and isolation of the fungus, but this method may require several days. Direct microscopic examination, although rapid, is relatively insensitive. Biochemical and immunodiagnostic rapid tests are also used. However, all of these methods have limitations that may hinder final diagnosis. The increasing incidence of fungal infections has focused attention on tools for rapid and accurate diagnosis using molecular biological techniques. Currently, PCR-based methods, particularly nested, multiplex and real-time PCR, provide both high sensitivity and specificity. In the present study, we evaluated a nested PCR targeting the gene encoding the ITS-1 and ITS-2 regions of rDNA in samples from a cohort of patients diagnosed with cryptococcosis. The results showed that in our hands, this Cryptococcus nested PCR assay has 100% specificity and 100% sensitivity and was able to detect until 2 femtograms of Cryptococcus DNA

Validation and clinical application of a nested PCR for paracoccidioidomycosis diagnosis in clinical samples from Colombian patients

Paracoccidioidomycosis is a systemic and endemic mycosis, restricted to tropical and subtropical areas of Latin America. The infection is caused by the thermal dimorphic fungus Paracoccidioides brasiliensisand Paracoccidioides lutzii. The diagnosis of paracoccidioidomycosis is usually performed by microscopic examination, culture and immunodiagnostic tests to respiratory specimens, body fluids and/or biopsies; however these methods require laboratory personnel with experience and several days to produce a result. In the present study, we have validated and evaluated a nested PCR assay targeting the gene encoding the Paracoccidioides gp43membrane protein in 191 clinical samples: 115 samples from patients with proven infections other than paracoccidioidomycosis, 51 samples as negative controls, and 25 samples from patients diagnosed with paracoccidioidomycosis. Additionally, the specificity of the nested PCR assay was also evaluated using purified DNA isolated from cultures of different microorganisms (n= 35) previously identified by culture and/or sequencing. The results showed that in our hands, this nested PCR assay for gp43 protein showed specificity and sensitivity rates of 100%. The optimized nested PCR conditions in our laboratory allowed detection down to 1 fg of P. brasiliensisDNA