CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Pesquisas cientÃficas buscando a utilizaÃÃo de antÃgenos de Coccidioides posadasii sÃo comuns na literatura especializada, uma vez que, sÃo instrumentos relevantes para diagnÃstico e possÃvel efeito imunoprotetor em humanos. Diante do exposto, foi realizada caracterizaÃÃo bioquÃmica de antÃgeno protÃico oriundo de C. posadasii. busca ativa de casos humanos de coccidioidomicose no Estado do CearÃ, bem como, avaliaÃÃo da resposta imunolÃgica in vivo. Para concretizar tais objetivos, a caracterizaÃÃo bioquÃmica do antÃgeno foi realizada por meio de eletroforese (SDS-PAGE e 2D-PAGE), detecÃÃo de proteases, glicoproteÃnas e sequenciamento N-terminal e foram buscados, ativamente, quadros de coccidiodiomicose em trÃs caÃadores de tatu com a respectiva descriÃÃo clÃnica e avaliaÃÃo laboratorial. Ademais, foi descrito modelo murino de coccidioidomicose com testagem de possÃvel efeito imunoprotetor do antÃgeno in-house. Quanto a anÃlise bioquÃmica, observou-se delimitaÃÃo de bandas nas faixas de 45-67 kDa e 67-97 kDa (SDS-PAGE), detecÃÃo de glicoproteÃnas, proteases e sequenciamento N-terminal demonstrando serem as bandas uma β-glucosidase e uma glutamina sintetase. Nos casos clÃnicos, foram encontrados queixas pneumÃnicas, exame micolÃgico direto, cultivo de escarro, imunodifusÃo radial dupla (com antÃgeno in-house) e PCR positivos para C. posadasii. No modelo murino, o grupo infectado, na presenÃa de antÃgeno e adjuvante, apresentou, na anÃlise histopatolÃgica, menores alteraÃÃes pulmonares que os demais grupos, alÃm de maior estÃmulo linfÃide esplÃnico. No que concerne a dosagem de citocinas (IL-6, IL-12 e TNFα), nÃo se observou diferenÃa significativa entre os grupos, mas uma tendÃncia à resposta imunoprotetora. O grupo infectado, sem imunoproteÃÃo, apresentou maior perda ponderal. Na anÃlise macroscÃpica, o mÃximo comprometimento foi a presenÃa de 2 granulomas, neste Ãltimo grupo. Na anÃlise dos hemogramas, a sÃrie branca demonstrou maiores diferenÃas entre os grupos. Em conclusÃo, o antÃgeno in-house se mostrou tratar-se de uma β-glicosidase e uma glutamina sintetase que tenderam a estimular a imunidade celular em modelo murino. AlÃm disso, a descriÃÃo de casos humanos contribui para a difusÃo do diagnÃstico precoce da coccidioidomicose e requer investigaÃÃes laboratoriais complementares. / Scientific researches seeking to use Coccidioides posadasii antigens are common in the literature, as relevant instruments to diagnosis and possible immunoprotector effect in humans. Thus, we performed biochemical characterization of protein antigen derived from C. posadasii, an active search for human cases of coccidioidomycosis in CearÃ, as well as evaluation of immune response in vivo. To achieve these goals, the biochemical characterization of the antigen was performed by electrophoresis (SDS-PAGE and 2D-PAGE), detection of proteases, glycoproteins and N-terminal sequencing and we reported three human cases of coccidiodiomycosis in armadillo hunters with its clinical description and laboratory evaluation. Further, a murine model was described testing a possible imunoprotector effect with an in-house antigen. For biochemical analysis, it was observed delimitation bands in ranges 45-67 kDa and 67-97 kDa (SDS-PAGE), detection of glycoproteins, proteases and N-terminal sequencing demonstrating the bands being a β-glucosidase and a glutamine synthetase. In clinical cases, it was found pneumonic disease, direct mycological examination, sputum culture, double radial immunodiffusion (antigen in-house) and PCR positive for C. posadasii. In the murine model, the infected group, in the presence of antigen and adjuvant, showed, histologically, lung disorders smaller than the other groups, and increased splenic lymphoid stimulus. Regarding the cytokines (IL-6, IL-12 and TNF), there was no significant difference between groups, but a trend toward immunoprotective response. The infected group, without immunoprotection, showed greater weight loss. In the macroscopic analysis, the maximum commitment was the presence of 2 granulomas in the latter group. In the analysis of blood counts, the white run showed major differences between groups. In conclusion, the in-house antigen showed that it was a β-glucosidase and a glutamine synthetase which tended to stimulate cellular immunity in a murine model. Moreover, the description of human cases contributes to the spread of early diagnosis of coccidioidomycosis and requires additional laboratory investigations.

Coccidioidomycosis, or Valley Fever, is an infectious disease caused by inhalation of soil-dwelling fungus Coccidioides posadasii spores in the Lower Sonoran Life Zone (LSLZ) in Arizona. In the context of climate change, the habitat of environmentally-mediated infectious diseases, such as Valley Fever, are expected to change. Connections have been drawn between climate and Valley Fever infection. The operational scale of the organism is still unknown. Here, we use climatic variables, including precipitation, soil moisture, and temperature. We use PRISM precipitation and temperature data, and Moderate Resolution Imaging Spectroradiometer (MODIS) Normalized Difference Vegetation Index (NDVI) as a measure of soil moisture for the entire state of Arizona, divided into 126 primary care areas (PCA). These data are analyzed and regressed with Valley Fever incidence to determine the effects of climatic variability on disease distribution and timing. This study confirms that Valley Fever occurrence is clustered in the LSLZ. Seasonal Valley Fever outbreak was found to be variable year-to-year based on climatic variability. The inconclusive regression analyses indicate that the operational scale of Coccidioides is smaller than the PCA region. All variables are related to Valley Fever infection, but one variable was not found to hold more predictive power than others.

An epidemiological survey of Valley Fever (coccidioidomycosis) was conducted in dogs in Tucson and Phoenix. Dogs were tested for delayed type hypersensitivity (DTH) using coccidioidin, a reagent that is not commercially available and results of the skin tests were compared with corresponding serologic test results.

Dogs in the Tucson area with a known history of clinical Valley Fever were tested with two skin test reagents to determine their ability to detect delayed type hypersensitivity (DTH) to the Coccidioides spp. The reagents used were Spherusol, from Nielsen Biologicals, and coccidioidin, which is no longer commercially available. Skin tests were read 48 hours after placement and evaluated for erythema and/or induration.

Histopathology of a murine lymph node, 9 days post infection with Coccidioides. Magnification 10X

Rupture of a coccidioidal pulmonary cavity with subsequent pyopneumothorax is a rare clinical event, even in areas endemic for coccidioidomycosis. Our encounter with a patient diagnosed with this condition in northeast Tennessee serves notice to clinicians that coccidioidomycosis is indeed a traveling fungal disease, and practitioners must be alert to common and uncommon manifestations of infection associated with this fungus. A literature review pertaining to coccidioidal pyopneumothorax revealed that patients usually present with a recent onset of chest pain. Serologic testing and pleural fluid culture are highly useful, and management includes surgical intervention with or without antifungal therapy.

Diagnostic medical mycology has been slow to advance due to a lack of species specific antigens in organisms which cause serious diseases in man. Toward this end, an HPLC analysis was done of the following fungal antigens: histoplasmins HKC-43 and H-42, blastomycin KCB-26, an alkali-soluble, water soluble antigen from Blastomyces dermatitidis (b-ASWS), a coccidioidin prepared from a toluene lysate of the mycelial-arthroconidia phase of Coccidioides immitis, and an alkali-soluble, water-soluble antigen from Coccidioides immitis (c-ASWS). The HPLC survey included size-exclusion chromatography (SEC), ion exchange chromatography (HPIEC), and reversephase chromatography (RP). Resolution was poor with both SEC and HPIEC but was excellent with RP chromatography. The use of RP will allow sufficient separation for further antigenic and structural analysis.

It is important to realize that guidelines cannot always account for individual variation among patients. They are not intended to supplant physician judgment with respect to particular patients or special clinical situations. Infectious Diseases Society of America considers adherence to these guidelines to be voluntary, with the ultimate determination regarding their application to be made by the physician in the light of each patient's individual circumstances. Coccidioidomycosis, also known as San Joaquin Valley fever, is a systemic infection endemic to parts of the southwestern United States and elsewhere in the Western Hemisphere. Residence in and recent travel to these areas are critical elements for the accurate recognition of patients who develop this infection. In this practice guideline, we have organized our recommendations to address actionable questions concerning the entire spectrum of clinical syndromes. These can range from initial pulmonary infection, which eventually resolves whether or not antifungal therapy is administered, to a variety of pulmonary and extrapulmonary complications. Additional recommendations address management of coccidioidomycosis occurring for special at-risk populations. Finally, preemptive management strategies are outlined in certain at-risk populations and after unintentional laboratory exposure.

It is important to realize that guidelines cannot always account for individual variation among patients. They are not intended to supplant physician judgment with respect to particular patients or special clinical situations. Infectious Diseases Society of America considers adherence to these guidelines to be voluntary, with the ultimate determination regarding their application to be made by the physician in the light of each patient's individual circumstances. Coccidioidomycosis, also known as San Joaquin Valley fever, is a systemic infection endemic to parts of the southwestern United States and elsewhere in the Western Hemisphere. Residence in and recent travel to these areas are critical elements for the accurate recognition of patients who develop this infection. In this practice guideline, we have organized our recommendations to address actionable questions concerning the entire spectrum of clinical syndromes. These can range from initial pulmonary infection, which eventually resolves whether or not antifungal therapy is administered, to a variety of pulmonary and extrapulmonary complications. Additional recommendations address management of coccidioidomycosis occurring for special at-risk populations. Finally, preemptive management strategies are outlined in certain at-risk populations and after unintentional laboratory exposure.

Nikkomycin Z (NikZ) is a chitin synthase inhibitor with antifungal efficacy against Coccidioides spp. and other endemic fungi. Dogs suffer a rate and range of natural coccidioidomycosis similar to humans and were considered an excellent model for initially testing NikZ against naturally acquired disease. Twelve dogs with coccidioidal pneumonia that had been present for an average of three months were treated with 250 mg (5 - 15 kg) or 500 mg (> 15 - 30 kg) twice daily for 60 days. Nine dogs completed the course of treatment and seven dogs had improvement in disease based on radiographs, clinicopathological parameters, physical examination findings, and subjective assessment by the owners; three dogs had resolution or near resolution of disease. Based on this small study, NikZ shows efficacy to treat naturally acquired coccidioidomycosis and merits further development for trials in humans.