RESUMO
In the original scientific publication evaluating sunscreen methodologies, Garzarella and Caswell showed there to be no clinically significant or statistically significant difference in the average Sun Protection Factor (SPF) of a sunscreen formulation between any of three methodologies, Food and Drug Administration (FDA) Final Monograph, Australia/New Zealand, and European Cosmetics Association (COLIPA) International, suggesting that any differences in methodology were insignificant in the resulting SPF determined. These three major older methodologies have coalesced into two methodologies, 2011 FDA-Final Rule and ISO 24444, so that current sunscreen SPF testing is mostly 2011 FDA-Final Rule and ISO 24444. Another approach to evaluating the impact of methodological differences in sunscreen testing is to compare data on a control standard or reference sunscreen. If the difference between the two SPF values of P2 is statistically significant for the two different methodologies, then this would present evidence for a clinically significant difference in the SPF value between the two methodologies. For 2011 FDA-Final Rule, the expected SPF of P2 is 16.3 ± 3.43; for ISO 24444, the expected SPF of P2 is 16.1 ± 2.42. Using least squares average and standard error on 952 observations, the 2011 FDA-Final Rule SPF of P2 is 15.4 ± 0.12; using least squares average and standard error on 1,551 observations, the ISO 24444 SPF of P2 is 15.6 ± 0.10. The data described herein indicate no clinically significant nor statistically significant difference between the SPF average of P2 using the 2011 FDA-Final Rule methodology versus that using ISO 24444 methodology. Further statistical analysis indicates that the average SPF of P2 is independent of solar simulator type, time of year (month), age of subject, gender of subject, or Fitzpatrick Skin Phototype of subject. A statistically significant negative correlation was found between a subject's SPF of P2 and the subject's unprotected minimal erythemal dose. The implications of this relationship on SPF testing are explored.
Assuntos
Cosméticos , Fator de Proteção Solar , Humanos , Luz Solar , Protetores Solares , Raios Ultravioleta , Estados Unidos , United States Food and Drug AdministrationRESUMO
Recovery has come to mean living a life beyond mental illness, and recovery orientation is policy in many countries. The aims of this study were to investigate what staff say they do to support recovery and to identify what they perceive as barriers and facilitators associated with providing recovery-oriented support. Data collection included ten focus groups with multidisciplinary clinicians (n = 34) and team leaders (n = 31), and individual interviews with clinicians (n = 18), team leaders (n = 6) and senior managers (n = 8). The identified core category was Competing Priorities, with staff identifying conflicting system priorities that influence how recovery-oriented practice is implemented. Three sub-categories were: Health Process Priorities, Business Priorities, and Staff Role Perception. Efforts to transform services towards a recovery orientation require a whole-systems approach.
Assuntos
Atitude do Pessoal de Saúde , Transtornos Mentais/reabilitação , Reabilitação Psiquiátrica , Adulto , Grupos Focais , Custos de Cuidados de Saúde , Humanos , Serviços de Saúde Mental , Pessoa de Meia-Idade , Avaliação de Processos em Cuidados de Saúde , Papel Profissional , Pesquisa Qualitativa , Medicina Estatal , Reino Unido , Adulto JovemRESUMO
BACKGROUND: The purpose of these clinical trials was to determine if 300 W and 150 W xenon arc solar simulators (SSs) deliver the same sun protection factor (SPF) and UVA protection factor (PFA). METHODS: First, the SPF of the P7 control standard and of the P2 control standard was determined, testing 20 subjects using the method described in the Food and Drug Administration (FDA) Final Monograph and using 150 W and 300 W SSs. In the second clinical trial, the PFA of the Japanese Cosmetic Industry Association (JCIA) control standard and of the P2 control standard was determined, testing 10 subjects using the method described in the JCIA Technical Bulletin and using 150 W and 300 W SSs. RESULTS: The SPF values for P7 control standard determined using the 150 W and 300 W SSs were 4.54 ± 0.35 and 4.61 ± 0.32, respectively. The SPF values for P2 control standard determined using the 150 W and 300 W SSs were 17.0 ± 0.9 and 16.7 ± 0.9, respectively. The resultant PFA values for JCIA control standard determined using the 150 W and 300 W SSs were 4.06 ± 0.70 and 4.06 ± 0.70, respectively. The resultant PFA values for P2 control standard determined using the 150 W and 300 W SSs were 3.28 ± 0.25 and 3.44 ± 0.39, respectively. CONCLUSION: As the values are essentially identical for SPF and for PFA, the 150 W and 300 W SSs can be used interchangeably for SPF and PFA determinations.
Assuntos
Iluminação/normas , Teste de Materiais , Fator de Proteção Solar , Protetores Solares/normas , Raios Ultravioleta , Adulto , Benzofenonas , Cinamatos , Desenho de Equipamento , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Propiofenonas , Padrões de Referência , Luz Solar , Xenônio , Adulto Jovem , para-AminobenzoatosRESUMO
BACKGROUND: Patients with rheumatic diseases including rheumatoid arthritis (RA) are at increased risk for infections related to both the disease and its treatments. These include uncommonly reported infections due to histoplasmosis. METHODS: Medical record review of all patients with a diagnosis of RA who developed new histoplasmosis infection in an endemic region between Jan 1, 1998 and Jan 30, 2009 and who were seen at Mayo Clinic in Rochester, Minnesota was performed. RESULTS: Histoplasmosis was diagnosed in 26 patients. Most patients were on combination therapies; 15 were on anti-tumor necrosis factor (anti-TNF) agents, 15 on corticosteroids and 16 on methotrexate. Most received more than 6 months of itraconazole and/or amphotericin treatment. Two patients died of causes unrelated to histoplasmosis. Anti-TNF treatment was restarted in 4/15 patients, with recurrence of histoplasmosis in one. CONCLUSIONS: In this largest single center series of patients with RA and histoplasmosis in the era of immunomodulatory therapy, we found that most patients had longstanding disease and were on multiple immunomodulatory agents. Most cases were pulmonary; typical signs and symptoms of disease were frequently lacking.
Assuntos
Artrite Reumatoide/complicações , Histoplasmose/complicações , Corticosteroides/uso terapêutico , Adulto , Idoso , Antirreumáticos/uso terapêutico , Artrite Reumatoide/tratamento farmacológico , Artrite Reumatoide/epidemiologia , Artrite Reumatoide/microbiologia , Registros Eletrônicos de Saúde , Doenças Endêmicas , Feminino , Histoplasmose/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fator de Necrose Tumoral alfa/antagonistas & inibidoresRESUMO
The specific identification of filamentous bacteria in tissue sections can be difficult. The filamentous bacteria Actinomyces and Nocardia often produce similar host responses and single bacterial organisms seem morphologically similar; however, their differentiation may be clinically significant. In situ hybridization (ISH) may assist in the rapid and accurate identification of these microorganisms. In this study, DNA probes were directed against the variable regions of 16S ribosomal RNA genes of multiple Actinomyces and Nocardia spp. Probes were tested on 26 formalin-fixed, paraffin-embedded tissue specimens, each of which contained diagnostic foci of filamentous bacteria confirmed by both Gram and Grocott methenamine silver stains. On the basis of histology and clinical features, cases were classified as Actinomyces-related, n = 13 with 6 culture-proven cases and Nocardia-related, n = 13 with 11 culture-proven cases. Using this classification, all cases were assessed for cross-reactivity using other species-specific probes and probe specificity was determined. Overall, Gram and Grocott methenamine silver histochemical stains (100% sensitivity) were more sensitive than ISH (77% sensitivity for both Actinomyces and Nocardia probes). The slender caliber of filamentous bacteria was a limitation for ISH interpretation and necessitated careful examination of some slides. Probes demonstrated 100% specificity for identifying both species, 100% positive predictive value and 81% negative predictive value. No mixed infections were observed. This study demonstrates that ISH is highly specific for distinguishing between Actinomyces and Nocardia spp. in tissue sections. Although histochemical stains demonstrate greater sensitivity for organism detection, ISH is a rapid and specific technique that is especially useful for evaluating culture-negative or clinically unsuspected cases of filamentous bacterial infection.
Assuntos
Actinomyces/classificação , Infecções por Actinomycetales/diagnóstico , Hibridização In Situ/métodos , Nocardiose/diagnóstico , Nocardia/classificação , Patologia/métodos , Actinomyces/genética , DNA Bacteriano/genética , DNA Ribossômico/genética , Diagnóstico Diferencial , Formaldeído , Humanos , Nocardia/genética , Sondas de Oligonucleotídeos/genética , Inclusão em Parafina , RNA Ribossômico 16S/genética , Sensibilidade e EspecificidadeRESUMO
The sulfonamides were the first drugs with antituberculous effects. Their use was abandoned and basically forgotten with the advent of streptomycin and isoniazid combination treatment. There is a widespread belief, apparently based on testing a single isolate on questionable media, that Mycobacterium tuberculosis is resistant to trimethoprim-sulfamethoxazole (TMP-SMX). We saw a complex immunocompromised patient with tuberculosis who was initially treated with TMP-SMX without antituberculous drugs and defervesced on this treatment. An isolate of M. tuberculosis from this patient was found to be sensitive to TMP-SMX. We examined how frequently M. tuberculosis is sensitive to TMP-SMX. Isolates were tested for susceptibility to TMP-SMX on supplemented Middlebrook 7H10 plates. We found that 43 of 44 (98%) isolates of M. tuberculosis were susceptible to the combination of < or = 1 microg/ml of TMP and 19 microg/ml of SMX (< or = 1/19 microg/ml). Thus, the vast majority of our M. tuberculosis isolates were susceptible to TMP-SMX at an MIC similar to that for Mycobacterium kansasii, Mycobacterium marinum, and sensitive rapidly growing mycobacteria, organisms successfully treated with TMP-SMX as part of the treatment regimen. It is possible that TMP-SMX may be useful in treating patients with multiple-drug-resistant and extended drug-resistant tuberculosis. We feel that a clinical trial looking at the effectiveness of TMP-SMX as an antituberculous drug is worthwhile.
Assuntos
Anti-Infecciosos/farmacologia , Mycobacterium tuberculosis/efeitos dos fármacos , Combinação Trimetoprima e Sulfametoxazol/farmacologia , Farmacorresistência Bacteriana , Humanos , Testes de Sensibilidade Microbiana , Mycobacterium tuberculosis/crescimento & desenvolvimento , Tuberculose/tratamento farmacológicoRESUMO
Zygomycete infections can be devastating in immunocompromised hosts. Difficulties in the histopathologic differentiation of this class from other filamentous fungi (e.g., Aspergillus spp., Fusarium spp.) may lead to delays in diagnosis and initiation of appropriate treatment, thereby significantly affecting patient outcome. A real-time PCR assay was developed to detect species of the zygomycete genera Absidia, Apophysomyces, Cunninghamella, Mucor, Rhizopus, and Saksenaea in culture and tissue samples. Primers and fluorescence resonance energy transfer hybridization probes were designed to detect a 167-bp conserved region of the multicopy zygomycete cytochrome b gene. A plasmid containing target sequence from Mucor racemosus was constructed as a positive control. The analytical sensitivity of the assay is 10 targets/mul, and a specificity panel consisting of other filamentous fungi, yeasts (Candida spp.), and bacteria demonstrated no cross-reactivity in the assay. The clinical sensitivity and specificity of the assay from culture isolates were 100% (39/39) and 92% (59/64), respectively. Sensitivity and specificity determined using a limited number of fresh tissue specimens were both 100% (2/2). The sensitivity seen with formalin-fixed, paraffin-embedded tissues was 56% (35/62), and the specificity was 100% (19/19). The speed, sensitivity, and specificity of the PCR assay indicate that it is useful for the rapid and accurate detection of zygomycetes.
Assuntos
Fungos/isolamento & purificação , Micoses/diagnóstico , Reação em Cadeia da Polimerase/métodos , Citocromos b/genética , Primers do DNA/genética , Transferência Ressonante de Energia de Fluorescência , Proteínas Fúngicas/genética , Fungos/genética , Humanos , Micoses/microbiologia , Hibridização de Ácido Nucleico/métodos , Reação em Cadeia da Polimerase/normas , Padrões de Referência , Sensibilidade e Especificidade , Temperatura de TransiçãoAssuntos
Atitude do Pessoal de Saúde , Atitude Frente a Saúde , Transtornos Mentais , Serviços de Saúde Mental/normas , Avaliação das Necessidades/organização & administração , Recuperação de Função Fisiológica , Adulto , Inglaterra , Análise Fatorial , Feminino , Humanos , Masculino , Transtornos Mentais/psicologia , Transtornos Mentais/reabilitação , Pessoa de Meia-Idade , Pesquisa Metodológica em Enfermagem , Avaliação de Resultados em Cuidados de Saúde , Filosofia Médica , Projetos Piloto , Avaliação de Programas e Projetos de Saúde , Pesquisa Qualitativa , Autoimagem , Apoio Social , Medicina Estatal/normas , Inquéritos e QuestionáriosRESUMO
To our knowledge, an institutional review of systemic histoplasmosis has not been conducted in the United States since the major outbreaks in Indianapolis in 1978-4982. We conducted a retrospective review of all patients with systemic histoplasmosis diagnosed at Mayo Clinic over a 15-year period. The case definitions employed were based on an international consensus statement by the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group (EORTC/IFICG) and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (MSG). One hundred eleven patients with systemic histoplasmosis were identified between January 1, 1991, and December 31, 2005. Of these, 78 patients had disseminated histoplasmosis and 55 patients had Histoplasma capsulatum fungemia. The mean age of patients was 55 years, 66% were male, and 98% were white. Fifty-nine percent of patients were immunocompromised. Fever was the most frequently reported symptom (63%), followed by respiratory complaints (43%) and weight loss (37%). The peripheral white blood cell count was <3000 cells/mm in 28%, hemoglobin was <10 g/dL in 29%, and platelet count was <150,000 cells/mm in 41% of patients. Liver enzymes were elevated (alanine aminotransferase >60 U/L in 39%, aspartate aminotransferase >60 U/L in 27%), alkaline phosphatase was >200 U/L in 55%, and albumin was <3.5 g/dL in 70%. Serologic and histopathologic examinations were each positive in 75% of cases, Histoplasma urine antigen screening was positive in 80%, and H. capsulatum was culture positive in 84%. Forty-seven percent of patients were sequentially treated with an amphotericin B-containing product followed by itraconazole, 31% received itraconazole alone, and 7% received an amphotericin B-containing product only. Another 13% of patients did not receive antifungal treatment, and the remaining 2% did not have treatment data available. Sixty percent of patients required hospitalization, and in hospital mortality was 6% with a median survival time of 61 days. The relapse rate was 9%, with a median relapse-free survival of 857 days. Systemic histoplasmosis should be suspected in patients who have lived in endemic areas with fever, bone marrow suppression, and elevated hepatic enzymes, particularly if they are immunocompromised. Evaluation including a combination of Histoplasma serologic screening, urine antigen assay, and fungal culture will secure the diagnosis in most cases.
Assuntos
Histoplasmose/epidemiologia , Antifúngicos/uso terapêutico , Feminino , Fungemia/tratamento farmacológico , Fungemia/epidemiologia , Histoplasma/imunologia , Histoplasma/isolamento & purificação , Histoplasmose/diagnóstico , Histoplasmose/tratamento farmacológico , Hospitalização/estatística & dados numéricos , Humanos , Hospedeiro Imunocomprometido , Masculino , Pessoa de Meia-Idade , Minnesota/epidemiologia , Recidiva , Estudos RetrospectivosRESUMO
BACKGROUND: A 65-year-old woman with a history of liver and kidney transplantation presented with a pruritic and tender rash of 2-year duration on her left knee. METHODS: The patient had a physical examination, and multiple skin biopsies were performed. RESULTS: Examination showed a cluster of dome-shaped, violaceous papules. A skin biopsy specimen showed multiple granules containing basophilic material within the superficial to mid dermis. The grains were embedded in an eosinophilic substance and surrounded by granulomatous inflammation. A Gomori methenamine silver stain showed abundant septate hyphae, and cultures grew colonies of Microsporum canis. A diagnosis of dermatophytic granuloma, or pseudomycetoma, was made. After considering the patient's immune status, a prolonged course of fluconazole was initiated. CONCLUSION: Microsporum canis, a frequently encountered zoophilic dermatophyte associated with tinea capitis and tinea corporis, is rarely associated with a mycetoma-like presentation, grain formation, or systemic infection. The medical literature describes only six cases of M. canis causing mycetoma-like formations in immunocompromised and immunocompetent patients. This case report describes an unusual presentation of a common dermatophyte infection, and the literature review summarizes the importance of recognizing atypical presentations of dermatophyte infections. Such infections may warrant more aggressive treatment in immunocompromised patients.
Assuntos
Dermatomicoses/imunologia , Hospedeiro Imunocomprometido , Microsporum , Idoso , Antifúngicos/uso terapêutico , Dermatomicoses/tratamento farmacológico , Dermatomicoses/patologia , Feminino , Fluconazol/uso terapêutico , Granuloma/imunologia , Granuloma/patologia , Humanos , Imunossupressores/efeitos adversos , Transplante de Rim/imunologia , Transplante de Fígado/imunologiaRESUMO
Rhino-orbitocerebral mucormycosis (ROCM) caused by more common zygomycetes (e.g., Mucor) is known to cause rapidly fatal infections in immunocompromised patients. Apophysomyces elegans is an emerging zygomycete that has been reported to cause invasive cutaneous and rhino-orbitocerebral infections in immunocompetent individuals. Limited data exist describing the syndrome of ROCM caused by A. elegans. We describe a recent case and performed a comprehensive literature review to delineate the clinical characteristics of ROCM caused by A. elegans. Our case is a 50-year-old man with diabetes mellitus who presented with facial pain and right eye proptosis. Endoscopic sinus sampling revealed A. elegans. He was treated with liposomal amphotericin B and multiple debridements, with no disease on 1.5-year follow-up examination. Seven cases were identified on literature review, including the present case. Most patients (86%) were male, with a mean age of 40 years. Most patients (71%) did not have predisposing medical conditions. Three patients had predisposing head trauma. All presented with facial and/or periorbital pain. All had magnetic resonance imaging or computed tomography of the head showing intraorbital and/or sinus inflammation. Diagnosis was confirmed by histopathology and deep tissue culture in all cases. All patients required eye exenteration and extensive surgical debridement, in addition to intravenous amphotericin B. Six of the seven patients (86%) recovered. ROCM caused by A. elegans is rarely reported in the literature. Most such infections occurred in immunocompetent patients, often after facial trauma. Survival in ROCM caused by A. elegans is favorable in reported cases, with prompt surgical debridement and antifungal therapy.
Assuntos
Encefalopatias/etiologia , Mucorales/patogenicidade , Mucormicose/etiologia , Doenças Nasais/etiologia , Doenças Orbitárias/etiologia , Adulto , Anfotericina B/uso terapêutico , Antifúngicos/uso terapêutico , Encefalopatias/diagnóstico , Encefalopatias/microbiologia , Encefalopatias/terapia , Terapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mucorales/isolamento & purificação , Mucormicose/diagnóstico , Mucormicose/microbiologia , Mucormicose/terapia , Doenças Nasais/diagnóstico , Doenças Nasais/microbiologia , Doenças Nasais/terapia , Exenteração Orbitária , Doenças Orbitárias/diagnóstico , Doenças Orbitárias/microbiologia , Doenças Orbitárias/terapiaRESUMO
Pneumocystis jiroveci is an important cause of pneumonia in immunocompromised individuals. This organism cannot be cultured, and therefore, diagnosis relies on microscopic identification of the organism using stains or antibodies. Although simple, these tests are insensitive and require expertise for accurate interpretation. We developed a real-time polymerase chain reaction (PCR) assay that provides sensitive and objective detection of Pneumocystis from bronchoalveolar lavage fluid. Primers and fluorescence resonance energy transfer probes were developed that target the cdc2 gene of P. jiroveci. Assay sensitivity is 6 copies of target per microliter of sample. No cross-reactivity occurs with other pathogens, and the PCR assay has a 21% increase in clinical sensitivity as compared with Calcofluor white staining. The real-time PCR assay provides a sensitive, rapid, and objective method for the detection of Pneumocystis from bronchoalveolar lavage fluid.
Assuntos
Líquido da Lavagem Broncoalveolar/microbiologia , Pneumocystis carinii/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Benzenossulfonatos , Primers do DNA , DNA Fúngico/análise , Transferência Ressonante de Energia de Fluorescência , Corantes Fluorescentes , Proteínas Fúngicas/genética , Humanos , Pneumocystis carinii/genética , Sensibilidade e Especificidade , Coloração e Rotulagem/métodosRESUMO
Recent molecular studies have shown Mycobacterium porcinum, recovered from cases of lymphadenitis in swine, to have complete 16S rDNA sequence identity and >70% DNA-DNA homology with human isolates within the M. fortuitum third biovariant complex. We identified 67 clinical and two environmental isolates of the M. fortuitum third biovariant sorbitol-negative group, of which 48 (70%) had the same PCR restriction enzyme analysis (PRA) profile as the hsp65 gene of M. porcinum (ATCC 33776(T)) and were studied in more detail. Most U.S. patient isolates were from Texas (44%), Florida (19%), or other southern coastal states (15%). Clinical infections included wound infections (62%), central catheter infections and/or bacteremia (16%), and possible pneumonitis (18%). Sequencing of the 16S rRNA gene (1,463 bp) showed 100% identity with M. porcinum ATCC 33776(T). Sequencing of 441 bp of the hsp65 gene showed four sequevars that differed by 2 to 3 bp from the porcine strains. Clinical isolates were positive for arylsulfatase activity at 3 days, nitrate, iron uptake, D-mannitol, i-myo-inositol, and catalase at 68 degrees C. They were negative for L-rhamnose and D-glucitol (sorbitol). Clinical isolates were susceptible to ciprofloxacin, sulfamethoxazole, and linezolid and susceptible or intermediate to cefoxitin, clarithromycin, imipenem, and amikacin. M. porcinum ATCC 33776(T) gave similar results except for being nitrate negative. These studies showed almost complete phenotypic and molecular identity between clinical isolates of the M. fortuitum third biovariant D-sorbitol-negative group and porcine strains of M. porcinum and confirmed that they belong to the same species. Identification of M. porcinum presently requires hsp65 gene PRA or 16S rRNA or hsp65 gene sequencing.
Assuntos
Técnicas de Tipagem Bacteriana , Mycobacterium/classificação , Suínos/microbiologia , Animais , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Sequência de Bases , Chaperonina 60 , Chaperoninas/genética , DNA Ribossômico/análise , Humanos , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Mycobacterium/efeitos dos fármacos , Mycobacterium/genética , Mycobacterium/patogenicidade , Infecções por Mycobacterium/microbiologia , Infecções por Mycobacterium/fisiopatologia , Mycobacterium fortuitum/classificação , Mycobacterium fortuitum/efeitos dos fármacos , Mycobacterium fortuitum/genética , Mycobacterium fortuitum/patogenicidade , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
More yeasts and molds are now recognized to cause more human disease than ever before. This development is not due to a change in the virulence of these fungi, but rather to changes in the human host. These changes include immunosuppression secondary to the pandemic of HIV, the use of life-saving advances in chemotherapy and organ transplantation, and the use of corticosteroids and other immunosuppressive agents to treat a variety of diseases. Fungi that were once considered common saprophytes are now recognized as potential pathogens in these patients. This situation necessitates better communication than ever between the clinician, pathologist, and clinical mycologist to ensure the prompt and accurate determination of the cause of fungal diseases.
Assuntos
Doenças Transmissíveis Emergentes/epidemiologia , Doenças Transmissíveis Emergentes/imunologia , Micoses/epidemiologia , Micoses/imunologia , Doenças Transmissíveis Emergentes/microbiologia , Humanos , Micoses/microbiologiaRESUMO
Isoniazid, a first-line antibiotic used for the treatment of tuberculosis, is a prodrug that requires activation by the Mycobacterium tuberculosis enzyme KatG. The KatG(S315T) mutation causes isoniazid resistance while the KatG(R463L) variation is thought to be a polymorphism. Much of the work to date focused on isoniazid activation by KatG has utilized recombinant enzyme overexpressed in Escherichia coli. In this work, native KatG and KatG(S315T) were purified from M. tuberculosis, and KatG(R463L) was purified from Mycobacterium bovis. The native molecular weight, enzymatic activity, optical, resonance Raman, and EPR spectra, K(D) for isoniazid binding, and isoniazid oxidation rates were measured and compared for each native enzyme. Further, the properties of the native enzymes were compared and contrasted with those reported for recombinant KatG, KatG(S315T), and KatG(R463L) in order to assess the ability of the recombinant enzymes to act as good models for the native enzymes.
Assuntos
Substituição de Aminoácidos/genética , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Catalase/química , Catalase/genética , Catalase/isolamento & purificação , Mycobacterium bovis/enzimologia , Mutação Puntual/genética , Proteínas de Bactérias/metabolismo , Catalase/metabolismo , Farmacorresistência Bacteriana/genética , Isoniazida/metabolismo , Isoniazida/uso terapêutico , Cinética , Mycobacterium bovis/genética , Oxirredução , Ligação Proteica/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Análise Espectral , Especificidade por Substrato/genética , Tuberculose/dietoterapia , Tuberculose/microbiologia , Tuberculose/patologiaRESUMO
The isolation and identification of a novel, slow-growing, scotochromogenic, mycobacterial species is reported. A strain, designated MUP 1182T, was isolated from a cervical lymph node of a 3-year-old child. MUP 1182T is alcohol- and acid-fast, with a lipid pattern that is consistent with those of species that belong to the genus Mycobacterium. It grows slowly at 25-37 degrees C, but does not grow at 42 degrees C. The isolate was revealed to be biochemically distinct from previously described mycobacterial species: it has urease and Tween hydrolysis activities and lacks nitrate reductase, 3-day arylsulfatase and beta-glucosidase activities. Comparative 16S rDNA sequencing showed that isolate MUP 1182T represents a novel, slow-growing species that is related closely to Mycobacterium lentiflavum and Mycobacterium simiae. On the basis of these findings, the name Mycobacterium parmense sp. nov. is proposed, with MUP 1182T (=CIP 107385T=DSM 44553T) as the type strain.
Assuntos
Linfonodos/microbiologia , Mycobacterium/classificação , Arilsulfatases/metabolismo , Técnicas de Tipagem Bacteriana , Pré-Escolar , DNA Bacteriano/química , DNA Bacteriano/isolamento & purificação , DNA Ribossômico/química , Genes de RNAr , Humanos , Lipídeos/análise , Dados de Sequência Molecular , Mycobacterium/citologia , Mycobacterium/isolamento & purificação , Mycobacterium/fisiologia , Nitrato Redutase , Nitrato Redutases/metabolismo , Filogenia , Polissorbatos/metabolismo , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Homologia de Sequência , Coloração e Rotulagem , Temperatura , Urease/metabolismo , beta-Glucosidase/metabolismoRESUMO
Mycobacterium celatum is a recently described organism. Herein we describe a case of M. celatum lymphadenitis in an immunocompetent child and the first reported case of a M. celatum infection (lung abscess) in a transplant recipient. A literature review identified 19 other cases of M. celatum infection. Fifteen occurred in patients with the acquired immunodeficiency syndrome. Of these, nine were disseminated and six were localized (primarily to the lungs). The remaining 4 patients were immunocompetent and had localized infection (pneumonitis or lymphadenitis). Diagnosis of M. celatum infection can be challenging as M. celatum can cause false-positive results with the current version of the Amplified Mycobacterium tuberculosis Direct Test (Gen-Probe, San Diego, CA.). Definitive identification is available by DNA sequencing or high-performance liquid chromatography. M. celatum can cause infection in immunocompromised and immunocompetent hosts.
Assuntos
Mycobacterium tuberculosis/isolamento & purificação , Micobactérias não Tuberculosas/isolamento & purificação , Tuberculose Pulmonar/microbiologia , Infecções Oportunistas Relacionadas com a AIDS/diagnóstico , Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Adulto , Pré-Escolar , Doenças Transmissíveis Emergentes/microbiologia , DNA Bacteriano/análise , Diagnóstico Diferencial , Reações Falso-Positivas , Feminino , Humanos , Masculino , Testes de Sensibilidade Microbiana , Infecções por Mycobacterium/diagnóstico , Infecções por Mycobacterium/microbiologia , Tuberculose Pulmonar/diagnósticoRESUMO
Described herein is our experience with the MicroSeq D2 large-subunit rDNA sequencing kit for the identification of filamentous fungi encountered in the mycology laboratory at the Mayo Clinic. A total of 234 filamentous fungi recovered from clinical specimens were used in the evaluation. All were identified by using phenotypic characteristics as observed macroscopically and microscopically on any medium or a combination of media, which included Sabouraud's dextrose, inhibitory mold, cornmeal, Czapek-Dox, potato dextrose, and V8 juice agars; all isolates were sequenced using the MicroSeq D2 large-subunit rDNA sequencing kit. Of the of 234 isolates, 158 were correctly identified to the appropriate genus or genus and species by using nucleic acid sequencing. Sequences for 70 (29.9%) of the isolates (27 genera) were not included in the MicroSeq library. Of the 80 dematiaceous and 154 hyaline fungi sequenced, 65 and 51.2%, respectively, gave results concordant with those determined by phenotypic identification. Nucleic acid sequencing using the MicroSeq D2 large-subunit rDNA sequencing kit offers promise of being an accurate identification system; however, the associated library needs to include more of the clinically important genera and species.