Design, Synthesis and Anticandidal Evaluation of Indazole and Pyrazole Derivatives.

Candidiasis, caused by yeasts of the genus Candida, is the second cause of superficial and mucosal infections and the fourth cause of bloodstream infections. Although some antifungal drugs to treat candidiasis are available, resistant strains to current therapies are emerging. Therefore, the search for new candicidal compounds is certainly a priority. In this regard, a series of indazole and pyrazole derivatives were designed in this work, employing bioisosteric replacement, homologation, and molecular simplification as new anticandidal agents. Compounds were synthesized and evaluated against C. albicans, C. glabrata, and C. tropicalis strains. The series of 3-phenyl-1H-indazole moiety (10a-i) demonstrated to have the best broad anticandidal activity. Particularly, compound 10g, with N,N-diethylcarboxamide substituent, was the most active against C. albicans and both miconazole susceptible and resistant C. glabrata species. Therefore, the 3-phenyl-1H-indazole scaffold represents an opportunity for the development of new anticandidal agents with a new chemotype.

Synthesis and Biological Evaluation of 2H-Indazole Derivatives: Towards Antimicrobial and Anti-Inflammatory Dual Agents.

Indazole is considered a very important scaffold in medicinal chemistry. It is commonly found in compounds with diverse biological activities, e.g., antimicrobial and anti-inflammatory agents. Considering that infectious diseases are associated to an inflammatory response, we designed a set of 2H-indazole derivatives by hybridization of cyclic systems commonly found in antimicrobial and anti-inflammatory compounds. The derivatives were synthesized and tested against selected intestinal and vaginal pathogens, including the protozoa Giardia intestinalis, Entamoeba histolytica, and Trichomonas vaginalis; the bacteria Escherichia coli and Salmonella enterica serovar Typhi; and the yeasts Candida albicans and Candida glabrata. Biological evaluations revealed that synthesized compounds have antiprotozoal activity and, in most cases, are more potent than the reference drug metronidazole, e.g., compound 18 is 12.8 times more active than metronidazole against G. intestinalis. Furthermore, two 2,3-diphenyl-2H-indazole derivatives (18 and 23) showed in vitro growth inhibition against Candida albicans and Candida glabrata. In addition to their antimicrobial activity, the anti-inflammatory potential for selected compounds was evaluated in silico and in vitro against human cyclooxygenase-2 (COX-2). The results showed that compounds 18, 21, 23, and 26 display in vitro inhibitory activity against COX-2, whereas docking calculations suggest a similar binding mode as compared to rofecoxib, the crystallographic reference.

Biochemical characterization of a bacteriocin-like inhibitory substance produced by Enterococcus faecium MXVK29, isolated from Mexican traditional sausage.

BACKGROUND: Enterococci are lactic acid bacteria that can produce bacteriocins, which may offer an additional hurdle to control the growth of food-borne pathogens; moreover, these bacteriocins may have great potential as natural biopreservatives. The aim of this work was to characterize a bacteriocin-like inhibitory substance (BLIS) with antilisterial activity produced by an enterococcal strain. RESULTS: The bacteriogenic strain was isolated from Mexican fermented sausages and identified as Enterococcus faecium with 99% sequence similarity. Maximal activity was detected at 16 h, where bacterial growth was in middle of the stationary phase. The producer strain was not inhibited by its own antimicrobial peptide. BLIS showed a strong anti-Listeria activity and was inactivated by proteinase K. Heating (121 °C for 15 min) induced some inactivation, but thermotolerance was higher at acid pH values. The yield obtained with a pH-mediated purification process was 32.7%, showing a band with an estimated molecular weight of 3.5 kDa. Automated N-terminal Edman degradation showed the following sequence: YYGNGVTCGSHHCSVD. CONCLUSION: Biochemical characteristics of BLIS produced by E. faecium MXVK29 suggested that it belongs to Class IIa of the Klaenhammer classification and could be considered as a natural food preservative, although further studies need to be performed.

Citometría de fuujo: vínculo entre la investigación básica y la aplicación clínica

La citometría de flujo es un método analítico que permite la medición rápida de ciertas características físicas y químicas de células o partículas suspendidas en líquido que producen una señal de forma individual al interferir con una fuente de luz. Una de las características analíticas más importantes de los citómetros de flujo es su capacidad de medir múltiples parámetros celulares, como el tamaño, forma y complejidad y, por supuesto, cualquier componente celular o función que pueda ser marcada con un fluorocromo. Las aplicaciones más relevantes de la citometría de flujo en la práctica médica se relacionan con la hematología e inmunología clínicas, midiendo parámetros como número y clasificación de células sanguíneas. Esta técnica es empleada también en el conteo de subpoblaciones de linfocitos en pacientes con el virus de la inmunodeficiencia humana, así como la caracterización de leucemias agudas y síndromes linfoproliferativos crónicos, entre otros padecimientos. En los últimos 20 años, el análisis de enfermedades pulmonares de origen inmunológico por citometría de flujo ha jugado un papel importante en el entendimiento y diagnóstico de enfermedades como sarcoidosis, neumonía eosinofílica o neumonitis por hipersensibilidad. Las aplicaciones de la citometría de flujo son numerosas, lo cual ha permitido el empleo de estos instrumentos de manera amplia en los campos, tanto de la investigación biológica como médica. Esta revisión brinda un panorama general de los principios básicos de la citometría de flujo y la muestra como una herramienta reproducible y aplicable a una gran variedad de campos médicos, así como su empleo en el campo de las enfermedades pulmonares.

Flow cytometry is an analytical method that allows the rapid measurement of certain physical and chemical characteristics of cells or particles suspended in liquid and produce signals when they pass individually through a beam of light. An important analytical feature of flow cytometers is their ability to measure multiple cellular parameters such as cell size, shape and internal complexity and, of course any cell component or function that can be detected by a fluorescent dye. The most prominent uses of flow cytometry in medical practice are in the related fields of laboratory hematology and clinical immunology, for a variety of tasks involving blood cell counting and classification. This technique is also used for counting lymphocyte subpopulations in patients with HIV, characterization of acute leukemias and chronic lymphomas between other diseases. Over the last 20 years, analysis of immunologica lung diseases by flow cytometry has played a major role in the understanding and as tool of diagnosis, such as sarcoidosis, eosinophilic pneumonia or hypersensitivity pneumonitis. So the applications of flow cytometry are numerous, and this has lead to the widespread use of this instruments in biological research and medical fields. Overall this review shows a brief overview of basic principles of and shows this as a reproducible tool applicable to a wide range of medical approaches as well as its use in lung diseases field.

Antibody response in mice immunized by mucosal routes with formalin-inactivated enteropathogenic Escherichia coli (EPEC) strains.

Secretory and systemic antibody response in mice against enteropathogenic Escherichia coli (EPEC) was evaluated. Groups of mice were immunized with formalin inactivated EPEC 0127:H6 strain by intranasal, peroral, intragastric and intrarectal route, with and without cholera toxin (CT) used as mucosal adjuvant. Mice immunized subcutaneously and a non treated control group were included. Other groups of mice were immunized intranasally with different EPEC strains and a non pathogenic E. coli K12 strain. Antibody response tested by ELISA assay showed that specific anti EPEC 0127:H6 antibodies were induced in serum by intranasal, subcutaneous and intragastric routes. A strong increase of antibody response against EPEC 0127:H6 strain was observed in saliva after intranasal delivery, while a lower response was detected by peroral and intrarectal immunization. Only the intranasal route increased IgA anti EPEC 0127:H6 antibody titers in feces. Specific and cross reactive antibodies to EPEC 0127:H6 were seen in mice immunized intranasally with different EPEC strains. Some control mice showed a background of anti EPEC 0127:H6 antibodies in feces. CT had a negative effect as adjuvant. We showed that nasal mucosa rendered the strongest antibody response in serum and secretions. These results might contribute to optimize the protective effect of enteric vaccines against infections associated to EPEC.