Non-Coding RNAs are Differentially Expressed by Nocardia brasiliensis in Vitro and in Experimental Actinomycetoma.

INTRODUCTION: Nocardia spp. are common soil-inhabiting bacteria that frequently infect humans through traumatic injuries or inhalation routes and cause infections, such as actinomycetoma and nocardiosis, respectively. Nocardia brasiliensis is the main aetiological agent of actinomycetoma in various countries. Many bacterial non-coding RNAs are regulators of genes associated with virulence factors. OBJECTIVE: The aim of this work was to identify non-coding RNAs (ncRNAs) expressed during infection conditions and in free-living form (in vitro) in Nocardia brasiliensis. METHODS AND RESULT: The N. brasiliensis transcriptome (predominately < 200 nucleotides) was determined by RNA next-generation sequencing in both conditions. A total of seventy ncRNAs were identified in both conditions. Among these ncRNAs, 18 were differentially expressed, 12 were located within intergenic regions, and 2 were encoded as antisense of 2 different genes. Finally, 10 of these ncRNAs were studied by rapid amplification of cDNA ends and/or quantitative reverse transcription polymerase chain reaction. Interestingly, 3 transcripts corresponded to tRNA-derived fragments (tRNAsCys, Met, Thr), and one transcript was overlapped between an intergenic region and the 5´end of the 23S rRNA. Expression of these last four transcripts was increased during N. brasiliensis infection compared with the in vitro conditions. CONCLUSION: The results of this work suggest a possible role for these transcripts in the regulation of virulence genes in actinomycetoma pathogenesis.

[Update on the epidemiology of mycetoma in Mexico. A review of 3933 cases]. / Actualización de la epidemiología del micetoma en México. Revisión de 3,933 casos.

UNLABELLED: Mycetoma is one of the most frequent chronic subcutaneous infections in many tropical and subtropical regions. OBJECTIVE: To update the epidemiological data of mycetoma cases in Mexico. METHOD: A survey in the main mycological diagnosis centers in this country was performed. Each mycologist was requested for number of diagnosed mycetoma cases, age, sex, occupation, geographic origin, type of mycetoma, and etiological agents. RESULTS: Until 2012, we have registered 3,933 cases in the last 54 years. Sex distribution corresponds to 75.6% for men and 24.4% for women. In 75.72% is present in adults between 16-50 years old. The predominant work group of patients is farmers (58.41%) followed by housewives (21.79%). Most of patients come from Jalisco, Morelos, Nuevo Leon, Guerrero, Veracruz and Michoacan states. The most affected body areas are limbs (60.29%) and trunk (19.76%). Actinomycetoma has a frequency of 96.52%, and the commonest etiological agent is Nocardia brasiliensis (65.58%). Eumycetoma (3.48%) is mainly caused by Madurella grisea (28.47%) and M. mycetomatis (26.28%). CONCLUSIONS: Mycetoma is an under-diagnosed pathology representing a health problem in rural regions and must be attended with more interest by the health institutions.

Pneumocystosis in a patient with Crohn's disease treated with combination therapy with adalimumab.

Pneumocystis jirovecii pneumonia (PCP) is a potential complication of immunosuppression. Crohn's disease (CD) is an immune granulomatous disorder characterized by transmural inflammation that can affect any part of the gastrointestinal tract. Its treatment is based on steroids and immunosuppressants but in non-responders, biologic compounds such as anti-tumor necrosis factor alpha (TNF) antibodies have been used. Neutralization of TNF causes a decrease in the inflammatory response but increases susceptibility to opportunistic infections such as fungal infections. We report a young male with chronic diarrhea, fever and weight loss who was diagnosed with CD and began conventional treatment with immunosuppressants, but due to lack of response after several weeks, biologic therapy with adalimumab was initiated. Seven weeks later he developed persistent fever and upper respiratory symptoms. After chest CT, bronchoscopy and bronchial lavage, P. jirovecii was identified by silver staining and confirmed by immunofluorescence. To our knowledge this is the second case of pneumocystosis associated with the use of adalimumab in CD and the first reported Mexican case confirmed by microbiological and immunological studies in this setting.

Hepatotoxicity from ingestion of wild mushrooms of the genus Amanita section Phalloideae collected in Mexico City: two case reports.

We present two cases of acute liver injury resulting from consumption of wild mushrooms. The first case was a male who developed acute hepatitis after ingestion of diverse mushrooms including Amanita species. His clinical course was favorable with complete recovery of liver function. The second case was a male who developed acute liver failure (ALF) after ingestion of Amanita bisporigera. He required MARS therapy as a bridge to liver transplantation but transplantation was not performed because he succumbed to multiorgan failure. There are few trials demonstrating the efficacy of the different treatments for mushroom poisoning. These cases demonstrate that the consumption of wild mushrooms without proper knowledge of toxic species represents a serious and under recognized health problem.

Pathogenesis of dermatophytosis and tinea versicolor.

Dermatophytoses are infections caused by keratinophilic fungi known as dermatophytes. Several steps are required for infection to take place: contact, adherence, and invasion of keratin layers. The severity of the infection depends on the type of agent, environmental factors, and the host immunologic status. Tinea versicolor is caused by the Malassezia spp yeasts, which are microorganisms that belong to normal biota in seborrheic areas, but some contributing factors, such as the application of oily preparations, creams, an increase in ambient humidity, corticosteroid abuse, or genetic predisposition can induce its overgrowth in both filamentous and yeast structures. Exposure to sunlight stimulates the production of azelaic acid, which causes the appearance of hypopigmented spots. Currently, there is no scientific explanation for hyperpigmented lesions.

[Antifungal resistance: an emerging problem in Mexico]. / La resistencia a los antifúngicos: un problema emergente en México.

BACKGROUND: An increase in mycosis associated with therapeutic failure has been observed worldwide. The dearth of data in Mexico led us to study antifungal resistance. MATERIAL AND METHODS: Seventy six isolates of patients from the Hospital de Especialidades, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social were included: 36 with dermatophytoses and 40 with candidiasis. Dermatophytes were assesed using the E-test method and Candida spp. using the broth microdilution method. Antifungal drugs included itraconazole, ketoconazole and fluconazole for dermatophytes; in addition, voriconazole and amphotericin B were used to treat yeasts. RESULTS: From the 36 dermatophytes, seven isolates (19.4%) showed resistance to one or more antifungal drugs: three to Trichophyton rubrum, three to T. mentagrophytes and one to T. tonsurans. One T. rubrum isolate was resistant to the three azoles; the other six isolates were resistant to fluconazole only. From the 40 Candida isolates, 11 (27.5%) showed resistance: seven to ketoconazole and itraconazole; three only to itraconazole and one to ketoconazole. One C. glabrata isolate showed resistance to the four azoles. None of the yeasts showed resistance to amphotericin B. CONCLUSION: Therapeutic failure could be caused by drug resistance. In our study we found an antifungal resistance of 20% and 27.5% in dermatophytes and in yeasts respectively.

La resistencia a los antifúngicos: un problema emergente en México / Antifungal resistance: an emerging problem in Mexico

Antecedentes: Mundialmente se ha observado incremento en los casos de micosis asociada a falla terapéutica. Ante el desconocimiento real de este fenómeno en México, se decidió estudiar la resistencia a antifúngicos. Material y métodos: Se evaluaron 76 aislamientos de pacientes del Hospital de Especialidades, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social: 36 con dermatofitosis y 40 con candidiasis. Para dermatófitos se utilizó el método E-test® y para Candida spp. el método de microdilución en caldo. Los antimicóticos fueron itraconazol, ketoconazol y fluconazol para dermatófitos; además, voriconazol y anfotericina B para levaduras. Resultados: De los 36 dermatófitos, siete (19.4%) fueron resistentes a uno o más antifúngicos: tres Trichophyton rubrum, tres T. mentagrophytes y un T. tonsurans. Un T. rubrum mostró resistencia a los tres azoles; los seis aislamientos restantes fueron resistentes sólo a fluconazol. De los 40 aislamientos de Candida, 11 (27.5 %) mostraron resistencia: siete a ketoconazol e itraconazol; tres sólo a itraconazol y uno a ketoconazol. Un aislamiento de C. glabrata fue resistente a los cuatro azoles. Ninguna de las levaduras mostró resistencia a anfotericina B. Conclusiones: La falla terapéutica podría deberse a fenómenos de resistencia. En este trabajo se encontró una resistencia a antifúngicos de 20 y 27.5% en dermatófitos y levaduras, respectivamente.

BACKGROUND: An increase in mycosis associated with therapeutic failure has been observed worldwide. The dearth of data in Mexico led us to study antifungal resistance. MATERIAL AND METHODS: Seventy six isolates of patients from the Hospital de Especialidades, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social were included: 36 with dermatophytoses and 40 with candidiasis. Dermatophytes were assesed using the E-test method and Candida spp. using the broth microdilution method. Antifungal drugs included itraconazole, ketoconazole and fluconazole for dermatophytes; in addition, voriconazole and amphotericin B were used to treat yeasts. RESULTS: From the 36 dermatophytes, seven isolates (19.4%) showed resistance to one or more antifungal drugs: three to Trichophyton rubrum, three to T. mentagrophytes and one to T. tonsurans. One T. rubrum isolate was resistant to the three azoles; the other six isolates were resistant to fluconazole only. From the 40 Candida isolates, 11 (27.5%) showed resistance: seven to ketoconazole and itraconazole; three only to itraconazole and one to ketoconazole. One C. glabrata isolate showed resistance to the four azoles. None of the yeasts showed resistance to amphotericin B. CONCLUSION: Therapeutic failure could be caused by drug resistance. In our study we found an antifungal resistance of 20% and 27.5% in dermatophytes and in yeasts respectively.

[Micosis among five highly underprivileged Mexican communities]. / Micosis observadas en cinco comunidades mexicanas con alto grado de marginación.

BACKGROUND: In many small rural communities in Mexico, medical care is deficient, empirical or absent. OBJECTIVE: In order to improve health coverage in rural areas, the Mexican Institute of Social Security organizes Medical and Surgical Meetings of various specialties including Dermatology and Mycology (MSDM). These include visits to rural hospitals by dermatologists and a mycologist to care for underprivileged communities. In addition to taking samples, they establish the clinical diagnosis and indicate medical and/or surgical treatment, with follow-up visits when needed. MATERIAL AND METHODS: In 2004 and 2005, five MSDM in Chiapas (two), Puebla (one), Michoacán (one) and Oaxaca (one) were organized. Mycoses were within the first four skin pathologies detected. RESULTS: Direct examination with potassium hydroxide led to the diagnosis of mycosis and other skin diseases such as scabies, pediculosis or hair disorders. The sample cultures showed, in addition to common fungi as dermatophytes (Trichophyton rubrum, 19 cases), other uncommon fungal agents such as Trichosporon spp, Chrysosporium spp, Cryptococcus, Geotrichum spp and Aspergillus spp. Most of the candidiasis cases were caused by Candida parapsilosis (nine cases) followed by C. albicans (three cases).

[Antifungal and phagocytic immunostimulation treatment of chronic disseminated dermatophytosis]. / Dermatofitosis diseminada crónica asociada a fagocitosis deficiente tratada con antimicóticos e inmunoestimulación fagocitaria.

A 39 years old man with a long-standing disseminated dermatophytosis even with several antifungal treatments is presented. From lesions, Trichophyton rubrum var. typical downy, T. tonsurans and Candida albicans were isolated and showed sensivity to azolic compounds in vitro. The phagocytic activity in vitro compared with normal control was depressed. Treatment with itraconazole and immunomodulation using a bacterial antigen was indicated. During the last two years the patient has been clinical and mycologically healthy, and his phagocytosis activity has become normal. In patients with chronic and relapsing dermatophytosis, the immune response evaluation is recommended, and immunomodulation could be useful as a rational measure in patients with a particular immunodeficiency.

Dermatofitosis diseminada crónica asociada a fagocitosis deficiente tratada con antimicóticos e inmunoestimulación fagocitaria / Antifungal and phagocytic immunostimulation treatment of chronic disseminated dermatophytosis

Se presenta el caso de un hombre de 39 años de edad con dermatofitosis crónica diseminada. De las lesiones se aislaron Trichophyton rubrum var. vellosa típica, T. tonsurans y Candida albicans, todos sensibles in vitro a compuestos azólicos. La actividad fagocítica in vitro comparada con un control normal mostró deficiencia en la misma. Se indicó tratamiento con itraconazol e inmunoestimulación con antígeno bacteriano. Durante los dos últimos años el paciente se ha mantenido clínica y micológicamente sano y su actividad fagocitaria es normal. En los pacientes con dermatofitosis crónica recidivante es recomendable la valoración de la respuesta inmune y la inmunomodulación puede ser útil como parte del tratamiento.

A 39 years old man with a long-standing disseminated dermatophytosis even with several antifungal treatments is presented. From lesions, Trichophyton rubrum var. typical downy, T. tonsurans and Candida albicans were isolated and showed sensivity to azolic compounds in vitro. The phagocytic activity in vitro compared with normal control was depressed. Treatment with itraconazole and immunomodulation using a bacterial antigen was indicated. During the last two years the patient has been clinical and mycologically healthy, and his phagocytosis activity has become normal. In patients with chronic and relapsing dermatophytosis, the immune response evaluation is recommended, and immunomodulation could be useful as a rational measure in patients with a particular immunodeficiency.

Micosis observadas en cinco comunidades mexicanas con alto grado de marginación / Micosis among five highly underprivileged Mexican communities

Antecedentes. En las pequeñas comunidades rurales de México la atención médica es deficiente, empírica o inexistente. Objetivo. Para mejorar la cobertura de salud en esas zonas, el Instituto Mexicano del Seguro Social (IMSS) organiza Encuentros Médico- Quirúrgicos de varias especialidades, entre ellos, los encuentros Médico- Quirúrgicos en Dermatología (EM-QD). Estos consisten en visitas de trabajo a hospitales rurales por parte de un grupo de especialistas en dermatología y micología durante los cuales se atiende a personas de comunidades marginadas. Además del diagnóstico clínico y toma de muestras se proporciona el tratamiento médico o quirúrgico. Posteriormente se hacen visitas de seguimiento a los pacientes que lo necesiten. Material y métodos. Durante 2004 y 2005 se realizaron cinco EM-QD: Chiapas (dos), Puebla, Michoacán y Oaxaca. Las micosis estuvieron entre las primeras cuatro causas de dermatosis. Resultados. El examen directo con hidróxido de potasio permitió diagnosticar micosis y otras patologías como escabiosis, pediculosis o alteraciones de pelo. El cultivo de las muestras demostró, además de los hongos habituales como los dermatofitos (Trichophyton rubrum 19 aislamientos), otros hongos poco habituales como causa de micosis cutáneas, entre ellos: Trichosporon spp., Chrysosporium spp., Cryptococcus spp., Geotrichum spp., y Aspergillus spp. Las candidosis en su mayoría fueron causadas por C. parapsilosis (9 casos) seguida en frecuencia por C. albicans (3 pacientes).

BACKGROUND: In many small rural communities in Mexico, medical care is deficient, empirical or absent. OBJECTIVE: In order to improve health coverage in rural areas, the Mexican Institute of Social Security organizes Medical and Surgical Meetings of various specialties including Dermatology and Mycology (MSDM). These include visits to rural hospitals by dermatologists and a mycologist to care for underprivileged communities. In addition to taking samples, they establish the clinical diagnosis and indicate medical and/or surgical treatment, with follow-up visits when needed. MATERIAL AND METHODS: In 2004 and 2005, five MSDM in Chiapas (two), Puebla (one), Michoacán (one) and Oaxaca (one) were organized. Mycoses were within the first four skin pathologies detected. RESULTS: Direct examination with potassium hydroxide led to the diagnosis of mycosis and other skin diseases such as scabies, pediculosis or hair disorders. The sample cultures showed, in addition to common fungi as dermatophytes (Trichophyton rubrum, 19 cases), other uncommon fungal agents such as Trichosporon spp, Chrysosporium spp, Cryptococcus, Geotrichum spp and Aspergillus spp. Most of the candidiasis cases were caused by Candida parapsilosis (nine cases) followed by C. albicans (three cases).

Cytokine production and lymphocyte proliferation in patients with Nocardia brasiliensis actinomycetoma.

IFN-gamma, TNF-alpha, IL-4, IL-10 and IL-12 concentrations in the supernatant of peripheral blood mononuclear cell (PBMC) cultures and the in vitro proliferation of PBMC were studied in 25 patients with actinomycetoma caused by Nocardia brasiliensis and in 10 healthy controls from endemic zones. Cell cultures were stimulated by a N. brasiliensis crude cytoplasmic antigen (NB) and five semi-purified protein fractions (NB2, NB4, NB6, NB8, and NB10) separated by isoelectric. Phytohemagglutinin (PHA) and purified protein derivative (PPD) of Mycobacterium tuberculosis were used as control antigens. Skin tests were performed by injecting 0.1 ml of candidin and PPD intradermally (ID). Patients showed a poor response to tuberculin, while their response to candidin was more than two fold greater than that observed in the controls. Cell proliferation showed no statistically significant differences in either group. IFN-gamma production was higher in the healthy controls than in the patients, whereas TNF-alpha secretion was slightly higher in the patients' cultures. IL-4 was detected in the patients' cultures but not in the controls. IL-10 and IL-12 were present at low concentrations in both groups. These results suggest that patients with actinomycetoma show normal antigen recognition, but with low IFN-gamma production, and higher concentrations of IL-4, IL-10 and TNF-alpha in the patients' PBMC cultures, indicating that they probably have a Th2 type of immune response.

Cefotaxima más amikacina asociadas a inmunomodulación en el tratamiento de actinomicetoma resistente a tratamiento convencional / Cefotaximine plus amikacin associated to immunomodulation in actinomycetoma treatment resistant to conventional therapy

El actinomicetoma es un síndrome de localización subcutánea de evolución crónica. Se presenta el caso de un paciente con micetoma abdominal causado por Nocardia brasiliensis, que había sido resistente a diferentes esquemas terapéuticos durante varios años y que, además, presentaba deficiencia en la función fagocitaria. En este caso, la curación se obtuvo con la administración de dos ciclos (con duración de 23 días cada uno); se administró cefotaxima (1g cada 8 horas) y amikacina (50 mg cada 12 horas). La inmunomodulación se llevó a cabo mediante la administración de una dosis semanal de levamisol de 300 mg, durante cuatro semanas, y la aplicación dos veces por semana de 1 ml de antígeno bacteriano a una concentración 600,000,000 de bacterias por ml durante 20 meses. Se discute la utilidad del antibiograma en estos pacientes y la importancia de la investigación de la función inmunológica en pacientes con resistencia a los tratamientos convencionales