Multilocus phylogeny of the Trichophyton mentagrophytes species complex and the application of matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) mass spectrometry for the rapid identification of dermatophytes.

Despite intensive studies of the Trichophyton mentagrophytes species complex, its taxonomy still causes confusion. In this study, more than 70 dermatophytes were analyzed based on nuc rDNA ITS1-5.8S-ITS2 (ITS), D1-D2 domains of nuc 28S rDNA (D1D2), and ß-tubulin gene (TUBB) sequences to clarify phylogenetic relationships in the complex. This demonstrated that strains of the complex were divided into three major lineages with high statistical support: (i) T. benhamiae and related species; (ii) T. simii and two related species, T. quinckeanum and T. schoenleinii; and (iii) T. mentagrophytes, T. interdigitale, and related species. The major lineages could be further divided into 18 phylogroups, representing either individual species or phylogenetically distinct groups within species. Among strains of T. benhamiae, African isolates American Type Culture Collection (ATCC) 28064 and 28065 formed a phylogenetically distinct phylogroup from their type strain and were considered a distinct species. Strains of T. mentagrophytes were divided into at least four phylogroups based on combined sequence analysis, but some phylogroups showed closer relationships to T. interdigitale, T. equinum, and T. tonsurans when compared by individual genes. This indicates that identifying those species with one gene could lead to incorrect results. For rapid identification of those dermatophytes, each phylogroup was tested by matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) mass spectrometry using a database with customized reference spectra of each phylogroup. This system was able to identify all the tested strains to species level with higher than 91% accuracy, except for strains of T. interdigitale. The three phylogroups of T. benhamiae were well distinguished from one another with high identification accuracy, whereas phylogroups of T. mentagrophytes were often cross-identified to one another or to T. interdigitale. Further research should improve identification accuracy for some species, but the results suggested that MALDI-TOF MS could be a rapid and efficient identification tool for closely related dermatophytes in the T. mentagrophytes species complex.

Characterization of the medically important yeast Trichosporon mucoides and its close sister Trichosporon dermatis by traditional and advanced technologies.

Trichosporon dermatis is a causative agent of several mycoses in immunocompromised patients but is often misidentified as Trichosporon mucoides due to their phenotypic resemblance. In order to evaluate the current identification keys for these species and to develop a rapid and reliable identification method, 11 strains of these yeasts were fully characterized in this study by traditional and advanced technologies. DNA sequences of the internal transcribed spacer (ITS), IGS1, and D1/D2 regions identified six of the yeasts as T. dermatis that were previously known as T. mucoides, including ATCC 204094 that has been used as the quality-control strain of T. mucoides for the VITEK 2 system and other commercial yeast identification kits. These two species could not be differentiated reliably by any previously known phenotypic keys for the species, such as growth patterns on ethylamine, phloroglucinol and tyramine, or by the VITEK 2 system. On the other hand, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) proved to be a rapid and reliable identification tool for the two closely related yeasts. With newly added superspectra from fully authenticated reference strains, the VITEK MS system using MALDI-TOF MS successfully separated strains of T. dermatis and T. mucoides at a similarity level of approximately 67 % for the mass spectra data, and could identify these strains at the species level with 100 % accuracy in repeated tests. Furthermore, the in vitro susceptibility results indicated that itraconazole, posaconazole and voriconazole were more effective against both T. mucoides and T. dermatis than the other antifungal agents tested in this study.

Goma sifilítica intracraneal en un paciente adolescente VIH seronegativo: reporte de un caso / Intracranial syphilitic gummain an HIV negative adolescent patient: a case report

La sífilis es una enfermedad sistémica causada por la espiroqueta Treponema pallidum que compromete al sistema nervioso central en cualquier etapa y cuyas presentaciones clínicas se modificaron en las últimas décadas. Se presenta el caso de un varón de 16 años con antecedentes de conducta sexual de riesgo y sífilis hace 3 años, sin recibir tratamiento. Acude por cefalea y disminución de visión hace 4 meses. Evaluación inicial objetiva hemianopsia homónima derecha, edema papilar bilateral y retinopatía exudativa izquierda. Pruebas serológicas de sífilis reactivas, VIH: no reactivo. Resonancia magnética de encéfalo: proceso expansivo en lóbulo occipital izquierdo captador de contraste. Recibió tratamiento para sífilis por 3 días y por sospecha inicial de proceso neoproliferativo se realizó biopsia que evidenció proceso granulomatosocrónico sifilítico. En su reingreso, mostró mayor compromiso de agudeza y defecto del campo visual. Potenciales evocados visuales: ausente en ambos ojos. Se inició Penicilina G sódica por 4 semanas (por evidencia de mejoría imagenológica) asociado a corticoides, encontrando disminución serológica y discreta mejoría clínica al término del tratamiento. La neurosífilis gomatosa esuna presentación infrecuente de la sífilis terciaria debiéndose considerar como diagnóstico diferencial en lesiones expansivas intracraneales en individuos VIHseronegativos con serología de sífilis reactiva.

Syphilis is a systemic disease caused by the spirochete Treponema pallidum which can compromise the central nervous system at any stage and whose clinical presentations have been modified in recent decades. Were port the case of a 16 years old male with a 3-years history of sexual risk behavior and syphilis with no treatment who experienced headache and decreased vision for four months. Initial assessment showed right homonymous hemianopia, bilateral papilledema and left exudative retinopathy. Serological tests for syphilis: reactive HIV: non-reactive. Brain MRI revealed a single irregularly enhancing lesion in the left occipital lobe. He received treatment for syphilis for three days, and stereotactic biopsies were performed by the initial suspicion of brain tumour. Histological examination showed a chronic granulomatous consistent with neurosyphilis. In his read mission, greater affectation of acuity and visual field defect was evident. Visual evoked potentials: absent in both eyes. Penicillin G sodium was initiated for 4weeks (for evidence of improvement imagenological)associated with corticosteroids, showing serological and discrete clinical improvement at the end of treatment decreased. The gummatous neurosyphilisis a rare form of tertiary syphilis which ever considered as differential expansive intracranial lesions in HIV-seronegative individuals with reactive syphilis serology.

Managing particulates in cellular therapy.

he concept of particulates, while common to many in the pharmaceutical and blood transfusion disciplines, represents a distinct challenge in the field of cellular therapy. With newly discovered products advancing through clinical trials, the focus has shifted to ensuring products are manufactured in a reliable and safe manner. Given the unique manufacturing processes and resulting products (i.e. the cell being the active ingredient of the product), the way in which particulates are viewed and subsequently tested needs to be reviewed. No specific test or method for particulates will apply to all products, and guidance documents will be generated over time as more cell therapy products are approved. The details of the processes, testing methods used and acceptance criteria established for particulates will play a major role in generating the guidance documents. This will ultimately allow for the manufacture and administration of safe and effective products without thwarting advancement of the cellular therapy field. The intent of this review is to bring awareness to the topic of particulates with respect to cell therapy, and encourage a more open dialog and exchange of examples within the industry. We have reviewed the concept of particulates, where they originate and how they are introduced to cell therapy products, and the current methods available for their detection. We have also reviewed the relevance of current guidance documents and present potential strategies to move forward and address and control unwanted contaminating particulates in cell therapy products.

Cobertura de seguros de salud en relación con el uso de servicios médicos, condiciones de vida y percepción de la salud en Lima, Perú / Coverage of health insurance in relation with medical services use, living conditions and perceived health in Lima, Peru

Objetivos. Correlacionar la afiliación a seguros de salud y el uso de servicios médicos, condiciones de vida y la percepción de salud en pobladores de doce distritos de la ciudad de Lima, Perú. Material y métodos. Estudio descriptivo transversal poblacional, realizado entre enero y marzo de 2006 en doce distritos de Lima Metropolitana, usando una encuesta de salud de 72 preguntas. Resultados. Se entrevistó a 4355 pobladores, 48 por ciento de ellos no tenían seguro de salud. Comparado con personas que tenían seguros privados de salud, sanidades o seguridad social, los participantes sin seguro o coberturados por el seguro público (SIS) tuvieron menor nivel educativo y de vida (p menor que 0,001). Los encuestados sin seguro o con SIS, cuando perciben enfermedad recurren con mayor frecuencia a las farmacias que al médico (p menor que 0,001). El sistema público es el principal sistema de atención medica pues atiende al 62,1 por ciento de los encuestados, incluyendo encuestados con seguro social. La percepción de un estado de salud óptimo se asoció con la afiliación a un seguro privado (OR=2,5 IC95 por ciento=1,87- 3,38) y una edad menor de 35 años (OR=2,15; IC95 por ciento =1,70-2,58). Conclusiones. Los sistemas de seguro de salud en los distritos de Lima cubren diferentes segmentos socio-económicos, los asegurados de sistemas privados presentan mejores condiciones de vida y acceso a servicios médicos, con mejor percepción de su salud.

Objectives. Correlate affiliation to health insurance with the use of medical services, living conditions and perceivedhealth of residents in 12 districts of Lima. Material and methods. Descriptive cross-population study, conducted between January and March 2006 in 12 districts of Lima city, using a health survey of 72 questions. Results. 4355 interviews were obtained, 48 per cent had no health insurance. Compared with people who had private health insurance, social security or healing, participants without insurance or coverage by public insurance had lower level of education and living (p minor that 0.001). Participants without insurance or with public insurance when they perceive disease increased use pharmacies than doctor (p minor that 0.001). The public system is the principal care system that provides care to 62.1 per cent of the respondents, including people from other insurance systems. The perception of an optimal health condition was associated with having a private health system coverage (OR = 2.5; IC95 per cent = 1.87-3.38); and age less than 35 years (OR=2.1; IC95 per cent =1.70-2.58). Conclusions.Health insurance systems in Lima city cover different socio-economic stratums; people using private insurance systems have better living conditions, better access to health services and optimal perception of their health.

Persistent expression of PEDF in the eye using high-capacity adenovectors.

Ocular neovascularization, the growth of abnormal blood vessels in the eye, is a factor shared by the most common blinding diseases in developed countries. Pigment epithelium-derived factor (PEDF) is a potent antiangiogenic and neuroprotective protein that is normally produced in the eye. When delivered via an adenovector, PEDF can block the growth of new blood vessels and trigger the selective regression of abnormal vessels in animal models of ocular disease. Because of the absence of adenoviral genes, high-capacity (HC) adenovectors offer the potential for persistent transgene expression and enhanced tolerability. We have assessed the durability of PEDF expression and the induction of ocular inflammation following delivery of a PEDF-expressing HC adenovector compared to earlier generation vectors. The HC vector mediated prolonged PEDF expression in tissue-cultured pigmented epithelial cells and when delivered by intravitreal injection into the mouse eye. Delivery of first-generation adenovectors resulted in a dose-dependent increase in cytokine/chemokine gene expression, which correlated with the infiltration of inflammatory cells in the eye. In comparison, the levels of inflammatory gene expression and the intraocular infiltrate were substantially reduced following delivery of the HC vector. These results support the development of the HC adenovector gene delivery system for ocular disease.

Prevalencia y factores asociados a heridas punzo-cortantes en trabajadores de salud del primer nivel de atención. Dirección de Salud V Lima Ciudad, 2005 / Prevalence and factors associated with sharps injuries in health workers in primary care. Direccion de Salud V Lima Ciudad, 2005

Introducción: Los trabajadores de salud presentan un mayor riesgo de accidentes que involucran agentes biológicos, de tal suerte que la probabilidad de adquirir enfermedades infecciosas es alta. El número de trabajadores en el sector salud ha estado en crecimiento en el Perú durante los últimos 25 años, a pesar de ello los estudios sobre accidentes laborales en este grupo son escasos. Objetivos: Determinar la prevalencia de heridas punzo-cortantes entre trabajadores de salud del primer nivel de atención de la DISA V Lima Ciudad e identificar factores asociados a su ocurrencia. Material y métodos: Estudio analítico transversal desarrollado en la Dirección de Salud (DISA) V Lima Ciudad entre enero y mayo de 2005. La selección de la muestra fue mediante un muestreo sistemático. Se aplicó un cuestionario estandarizado auto-administrado luego de la aceptación del trabajador y firma del consentimiento informado. El análisis consistió en el cálculo de la prevalencia, estadística descriptiva e inferencial y estimación de OR e IC95%. Resultados: Se entrevistó un total de 203 trabajadores (28.5% varones), con una edad promedio de 45.9±8.3 años y tiempo de trabajo de 17.1±7.8 años. Durante los últimos cinco años el 34.0% refirió haber sufrido al menos una herida punzo-cortante (IC95% 27.5%-40.5%). Las ocupaciones con mayor prevalencia de heridas punzo-cortante fueron tecnólogos de laboratorio (50.0%) y técnicos de enfermería (40.3%). Los objetos más frecuentemente involucrados fueron agujas de inyectable (69.6%) y agujas de sutura (20.3%). La mayoría de las heridas (52.2%) fueron de profundidad intermedia (2 a 5mm, escaso sangrado); las heridas superficiales (menos de 2mm, sin sangrado) fueron el 40.6% y las profundas (5mm o más, sangrado profuso) el 7.2%...

Introduction: Health care workers (HCW) are at higher risk for accidents involving biological agents, thus the probability of acquiring infectious diseases is high. The number of HCW at the primary care level has been growing in Peru over the last 25 years, yet reports of work-related injuries in this group are scarce. Objective: To determine the prevalence of percutaneous injuries among HCW in primary care settings in Lima, Peru. Methods: A cross-sectional study was conducted at the Lima Department of Health between January and May 2005. HCW from various primary care settings were enlisted. A systematic sampling was performed. A standardized selfadministered questionnaire was given to participants to complete upon acceptance into the study. Data analysis consisted of calculation of the prevalence, descriptive and inferential statistics, and OR with 95% CI. Results: Of the 203 HCW surveyed, 28.5% were male, with an average age of 45.9±8.3 years and an health care employment duration of 17.1±7.8 years. Over the past five years, 34.0% suffered at least one percutaneous injury (95% CI 27.5%-40.5%). Occupations with the highest prevalence were laboratory technologists (50.0%) and practical nurses (40.3%). The objects most frequently involved in these injuries were injection needles (69.6%) and suture needles (20.3%). Most of the injuries (52.2%) were of intermediate depth (2 to 5mm in depth, little bleeding); superficial injuries (less than 2mm, no bleeding) accounted for 40.6% and 7.2% were deep injuries (5mm or more, profuse bleeding). 69.6% of HCW who suffered a percutaneous injury did not report it, 27.5% underwent HIV testing and 7.2% were tested for hepatitis B. Only one HCW received antiretroviral prophylaxis for HIV. Of the possible risk factors studied, only age was associated with a significant increase in the occurrence of percutaneous injuries (OR=6.0 for those under 45 years, 95% CI 1.8- 19.5)...

Periocular gene transfer of pigment epithelium-derived factor inhibits choroidal neovascularization in a human-sized eye.

Gene transfer provides a potential way to achieve sustained delivery of therapeutic proteins to the eye. Studies in rodents have suggested that periocular injection of adenoviral vectors containing expression cassettes for antiangiogenic proteins results in high intraocular levels of the proteins and suppression of choroidal neovascularization (CNV). However, the differences in size and scleral thickness between mouse and human eyes make it difficult to ascertain if periocular gene transfer is a feasible approach for treating human choroidal diseases. To address this issue, we tested the effect of periocular injection of an expression cassette for pigment epithelium-derived factor (PEDF) packaged in adenoviral vector (AdPEDF.11) in a CNV model in pigs, which have eyes that are very similar to humans in size and scleral thickness. Periocular injection of beta-galactosidase (AdLacZ.11) resulted in prominent transduction of periocular tissues, as was seen in mice. Periocular injection of AdPEDF.11 caused increased levels of PEDF in the choroid and significantly reduced the amount of CNV at rupture sites in Bruch's membrane. These data suggest that periocular gene transfer may be feasible for treatment of human choroidal diseases.

Periocular gene transfer of sFlt-1 suppresses ocular neovascularization and vascular endothelial growth factor-induced breakdown of the blood-retinal barrier.

Vascular endothelial growth factor (VEGF) is a critical stimulus for both retinal and choroidal neovascularization, and for diabetic macular edema. We used mouse models for these diseases to explore the potential of gene transfer of soluble VEGF receptor-1 (sFlt-1) as a treatment. Intravitreous or periocular injection of an adenoviral vector encoding sFlt-1 (AdsFlt-1.10) markedly suppressed choroidal neovascularization at rupture sites in Bruch's membrane. Periocular injection of AdsFlt-1.10 also caused significant reduction in VEGF-induced breakdown of the blood-retinal barrier, but failed to significantly inhibit ischemia-induced retinal neovascularization. Periocular delivery of an adenoviral vector encoding pigment epithelium-derived factor (PEDF), another secreted protein, resulted in high levels of PEDF in the retinal pigmented epithelium and choroid, but not in the retina. This may explain why periocular injection of AdsFlt-1.10 inhibited choroidal, but not retinal neovascularization. Periocular delivery offers potential advantages over other routes of delivery and the demonstration that sFlt-1 enters the eye from the periocular space in sufficient levels to achieve efficacy in treating choroidal neovascularization and retinal vascular permeability is a novel finding that has important clinical implications. These data suggest that periocular gene transfer of sFlt-1 should be considered for treatment of choroidal neovascularization and diabetic macular edema.