Multiphoton FLIM Analyses of Native and UVA-Modified Synthetic Melanins.

To better understand the impact of solar light exposure on human skin, the chemical characterization of native melanins and their structural photo-modifications is of central interest. As the methods used today are invasive, we investigated the possibility of using multiphoton fluorescence lifetime (FLIM) imaging, along with phasor and bi-exponential fitting analyses, as a non-invasive alternative method for the chemical analysis of native and UVA-exposed melanins. We demonstrated that multiphoton FLIM allows the discrimination between native DHI, DHICA, Dopa eumelanins, pheomelanin, and mixed eu-/pheo-melanin polymers. We exposed melanin samples to high UVA doses to maximize their structural modifications. The UVA-induced oxidative, photo-degradation, and crosslinking changes were evidenced via an increase in fluorescence lifetimes along with a decrease in their relative contributions. Moreover, we introduced a new phasor parameter of a relative fraction of a UVA-modified species and provided evidence for its sensitivity in assessing the UVA effects. Globally, the fluorescence lifetime properties were modulated in a melanin-dependent and UVA dose-dependent manner, with the strongest modifications being observed for DHICA eumelanin and the weakest for pheomelanin. Multiphoton FLIM phasor and bi-exponential analyses hold promising perspectives for in vivo human skin mixed melanins characterization under UVA or other sunlight exposure conditions.

In vivo multiphoton multiparametric 3D quantification of human skin aging on forearm and face.

Quantifying skin aging changes and characterizing its 3D structure and function in a non-invasive way is still a challenging area of research, constantly evolving with the development of imaging methods and image analysis tools. In vivo multiphoton imaging offers means to assess skin constituents in 3D, however prior skin aging studies mostly focused on 2D analyses of dermal fibers through their signals' intensities or densities. In this work, we designed and implemented multiphoton multiparametric 3D quantification tools for in vivo human skin pigmentation and aging characterization. We first demonstrated that despite the limited field of view of the technic, investigation of 2 regions of interest (ROIs) per zone per volunteer is a good compromise in assessing 3D skin constituents in both epidermis and superficial dermis. We then characterized skin aging on different UV exposed areas-ventral and dorsal forearms, face. The three major facts of aging that are epidermal atrophy, the dermal-epidermal junction (DEJ) flattening and dermal elastosis can be non-invasively quantified and compared. Epidermal morphological changes occur late and were only objectified between extreme age groups. Melanin accumulation in suprabasal layers with age and chronic exposure on ventral and dorsal forearms is less known and appears earlier. Superficial dermal aging changes are mainly elastin density increase, with no obvious change in collagen density, reflected by SHGto2PEF ratio and SAAID index decrease and ImbrN index increase on all skin areas. Analysis of the z-dermal distribution of these parameters highlighted the 2nd 20 µm thickness normalized dermal sub-layer, that follows the DEJ shape, as exhibiting the highest aging differences. Moreover, the 3D ImbrN index allows refining the share of photoaging in global aging on face and the 3D SAAID index on forearm, which elastin or fibrillar collagens densities alone do not allow. Photoaging of the temple area evolves as a function of chronic exposure with a more pronounced increase in elastin density, also structurally modified from thin and straight elastic fibers in young volunteers to dense and compact pattern in older ones. More generally, multiphoton multiparametric 3D skin quantification offers rich spatial information of interest in assessing normal human skin condition and its pathological, external environment or product induced changes.

In vivo melanin 3D quantification and z-epidermal distribution by multiphoton FLIM, phasor and Pseudo-FLIM analyses.

Characterizing melanins in situ and determining their 3D z-epidermal distribution is paramount for understanding physiological/pathological processes of melanin neosynthesis, transfer, degradation or modulation with external UV exposure or cosmetic/pharmaceutical products. Multiphoton fluorescence intensity- and lifetime-based approaches have been shown to afford melanin detection, but how can one quantify melanin in vivo in 3D from multiphoton fluorescence lifetime (FLIM) data, especially since FLIM imaging requires long image acquisition times not compatible with 3D imaging in a clinical setup? We propose an approach combining (i) multiphoton FLIM, (ii) fast image acquisition times, and (iii) a melanin detection method called Pseudo-FLIM, based on slope analysis of autofluorescence intensity decays from temporally binned data. We compare Pseudo-FLIM to FLIM bi-exponential and phasor analyses of synthetic melanin, melanocytes/keratinocytes coculture and in vivo human skin. Using parameters of global 3D epidermal melanin density and z-epidermal distribution profile, we provide first insights into the in vivo knowledge of 3D melanin modulations with constitutive pigmentation versus ethnicity, with seasonality over 1 year and with topical application of retinoic acid or retinol on human skin. Applications of Pseudo-FLIM based melanin detection encompass physiological, pathological, or environmental factors-induced pigmentation modulations up to whitening, anti-photoaging, or photoprotection products evaluation.

Simultaneous NAD(P)H and FAD fluorescence lifetime microscopy of long UVA-induced metabolic stress in reconstructed human skin.

Solar ultraviolet longwave UVA1 exposure of human skin has short-term consequences at cellular and molecular level, leading at long-term to photoaging. Following exposure, reactive oxygen species (ROS) are generated, inducing oxidative stress that might impair cellular metabolic activity. However, the dynamic of UVA1 impact on cellular metabolism remains unknown because of lacking adequate live imaging techniques. Here we assess the UVA1-induced metabolic stress response in reconstructed human skin with multicolor two-photon fluorescence lifetime microscopy (FLIM). Simultaneous imaging of nicotinamide adenine dinucleotide (NAD(P)H) and flavin adenine dinucleotide (FAD) by wavelength mixing allows quantifying cellular metabolism in function of NAD(P)+/NAD(P)H and FAD/FADH2 redox ratios. After UVA1 exposure, we observe an increase of fraction of bound NAD(P)H and decrease of fraction of bound FAD indicating a metabolic switch from glycolysis to oxidative phosphorylation or oxidative stress possibly correlated to ROS generation. NAD(P)H and FAD biomarkers have unique temporal dynamic and sensitivity to skin cell types and UVA1 dose. While the FAD biomarker is UVA1 dose-dependent in keratinocytes, the NAD(P)H biomarker shows no dose dependence in keratinocytes, but is directly affected after exposure in fibroblasts, thus reflecting different skin cells sensitivities to oxidative stress. Finally, we show that a sunscreen including a UVA1 filter prevents UVA1 metabolic stress response from occurring.

In vivo multiphoton imaging for non-invasive time course assessment of retinoids effects on human skin.

BACKGROUND: In vivo multiphoton imaging and automatic 3D image processing tools provide quantitative information on human skin constituents. These multiphoton-based tools allowed evidencing retinoids epidermal effects in the occlusive patch test protocol developed for antiaging products screening. This study aimed at investigating their relevance for non-invasive, time course assessment of retinoids cutaneous effects under real-life conditions for one year. MATERIALS AND METHODS: Thirty women, 55-65 y, applied either retinol (RO 0.3%) or retinoic acid (RA 0.025%) on one forearm dorsal side versus a control product on the other forearm once a day for 1 year. In vivo multiphoton imaging was performed every three months, and biopsies were taken after 1 year. Epidermal thickness and dermal-epidermal junction undulation were estimated in 3D with multiphoton and in 2D with histology, whereas global melanin density and its z-epidermal distribution were estimated using 3D multiphoton image processing tools. RESULTS: Main results after one year were as follows: a) epidermal thickening with RO (+30%); b) slight increase in dermal-epidermal junction undulation with RO; c) slight decrease in 3D melanin density with RA; d) limitation of the melanin ascent observed with seasonality and time within supra-basal layers with both retinoids, using multiphoton 3D-melanin z-epidermal profile. CONCLUSIONS: With a novel 3D descriptor of melanin z-epidermal distribution, in vivo multiphoton imaging allows demonstrating that daily usage of retinoids counteracts aging by acting not only on epidermal morphology, but also on melanin that is shown to accumulate in the supra-basal layers with time.

Cat-scratch Disease in the Pediatric Population: 6 Years of Evaluation and Follow-up in a Public Hospital in Chile.

BACKGROUND: Bartonella henselae is the etiologic agent of cat-scratch disease, which affects the entire world population. Due to how poorly characterized the child population is, the objective was to describe the clinical behavior of this disease in children younger than 16 years of age. METHODS: This was a descriptive study with prospective follow-up of all children younger than 16 years with clinical and serologic diagnoses between 2013 and 2018. RESULTS: There were 142 patients; of these, 55.6% had localized disease, 34.5% disseminated with hepatosplenic involvement and 9.8% had atypical disease. The cases of atypical disease were prolonged febrile illness, Parinaud syndrome, subacute bacterial endocarditis/glomerulonephritis and aseptic meningitis. Cervical lymphadenopathy was the most frequent, followed by inguinal adenopathy. There were no differences between the type of manifestation and laboratory values except for a higher erythrocyte sedimentation rate tendency in patients with disseminated and/or atypical disease. The serologic titers throughout were distributed as follows: titer of 1/1024, 71.7%; titer of 1/512, 10.4% and titer of 1/256, 17.9%. CONCLUSIONS: This is, to our knowledge, the largest series of cat-scratch disease published. We demonstrate that in our environment, the inguinal location is suggestive of this disease, the disseminated presentation, frequent, and that the titers are probably much higher than in other populations, which perhaps indicates the need to re-examine the cut-off point for positivity.

Imaging and quantifying drug delivery in skin - Part 2: Fluorescence andvibrational spectroscopic imaging methods.

Understanding the delivery and diffusion of topically-applied drugs on human skin is of paramount importance in both pharmaceutical and cosmetics research. This information is critical in early stages of drug development and allows the identification of the most promising ingredients delivered at optimal concentrations to their target skin compartments. Different skin imaging methods, invasive and non-invasive, are available to characterize and quantify the spatiotemporal distribution of a drug within ex vivo and in vivo human skin. The first part of this review detailed invasive imaging methods (autoradiography, MALDI and SIMS). This second part reviews non-invasive imaging methods that can be applied in vivo: i) fluorescence (conventional, confocal, and multiphoton) and second harmonic generation microscopies and ii) vibrational spectroscopic imaging methods (infrared, confocal Raman, and coherent Raman scattering microscopies). Finally, a flow chart for the selection of imaging methods is presented to guide human skin ex vivo and in vivo drug delivery studies.

Imaging and quantifying drug delivery in skin - Part 1: Autoradiography and mass spectrometry imaging.

In this two-part review we present an up-to-date description of different imaging methods available to map the localization of drugs on skin as a complement of established ex-vivo absorption studies. This first part deals with invasive methods which are grouped in two classes according to their underlying principles: i) methods using radioactivity such as autoradiography and ii) mass spectrometry methods such as MALDI and SIMS. For each method, a description of the principle is given along with example applications of imaging and quantifying drug delivery in human skin. Thanks to these techniques a better assessment of the fate of drugs is obtained: its localization on a particular skin structure, its potential accumulation, etc. A critical comparison in terms of capabilities, sensitivity and practical applicability is included that will help the reader to select the most appropriate technique depending on the particular problem to be solved.

[Occult bacteremia in Sótero del Río Hospital: Experience after pneumococcal conjugated vaccine]. / Bacteriemia oculta en niños atendidos en el Complejo Asistencial Dr. Sótero del Río: Experiencia post vacuna neumocóccica conjugada.

BACKGROUND: Occult bacteremia (OB) is one of the possible diagnoses of children younger than 3 years with fever without source in the emergency room. OBJECTIVE: describe OB in the era after introduction of pneumococcal vaccine in Chile. PATIENTS AND METHODS: Prospective descriptive review of data of children with possible OB diagnosis, referred from the emergency department between 2010-2013. RESULTS: Possible OB was diagnosed on 391 patients. 233 had focus, mainly respiratory virus and urinary tract infection. On 158 patients, probable BO was diagnosed, 20 had proven OB from which 15 had positive blood culture for Streptococcus pneumoniae. From these, 7 were fully or partially vaccinated. The serotype was identified on 14 cases: 6 were PCV10 vaccine serotypes (none of them vaccinated), 2 were serotype related (both partially vaccinated) and 6 were non vaccine serotypes (partially or totally vaccinated). DISCUSSION: It is necessary to improve diagnostic methods for respiratory viruses and urinary tract infections and try to expand coverage of pneumococcal conjugated vaccines in the pediatric population in order to reduce the risk of invasive pneumococcal disease in Chile.

Multicolor two-photon imaging of endogenous fluorophores in living tissues by wavelength mixing.

Two-photon imaging of endogenous fluorescence can provide physiological and metabolic information from intact tissues. However, simultaneous imaging of multiple intrinsic fluorophores, such as nicotinamide adenine dinucleotide(phosphate) (NAD(P)H), flavin adenine dinucleotide (FAD) and retinoids in living systems is generally hampered by sequential multi-wavelength excitation resulting in motion artifacts. Here, we report on efficient and simultaneous multicolor two-photon excitation of endogenous fluorophores with absorption spectra spanning the 750-1040 nm range, using wavelength mixing. By using two synchronized pulse trains at 760 and 1041 nm, an additional equivalent two-photon excitation wavelength at 879 nm is generated, and achieves simultaneous excitation of blue, green and red intrinsic fluorophores. This method permits an efficient simultaneous imaging of the metabolic coenzymes NADH and FAD to be implemented with perfect image co-registration, overcoming the difficulties associated with differences in absorption spectra and disparity in concentration. We demonstrate ratiometric redox imaging free of motion artifacts and simultaneous two-photon fluorescence lifetime imaging (FLIM) of NADH and FAD in living tissues. The lifetime gradients of NADH and FAD associated with different cellular metabolic and differentiation states in reconstructed human skin and in the germline of live C. Elegans are thus simultaneously measured. Finally, we present multicolor imaging of endogenous fluorophores and second harmonic generation (SHG) signals during the early stages of Zebrafish embryo development, evidencing fluorescence spectral changes associated with development.

Bacteriemia oculta en niños atendidos en el Complejo Asistencial Dr. Sótero del Río: experiencia post vacuna neumocóccica conjugada / Occult bacteremia in Sótero del Río Hospital: experience after pneumococcal conjugated vaccine

Background: Occult bacteremia (OB) is one of the possible diagnoses of children younger than 3 years with fever without source in the emergency room. Objective: describe OB in the era after introduction of pneumococcal vaccine in Chile. Patients and Methods: Prospective descriptive review of data of children with possible OB diagnosis, referred from the emergency department between 2010-2013. Results: Possible OB was diagnosed on 391 patients. 233 had focus, mainly respiratory virus and urinary tract infection. On 158 patients, probable BO was diagnosed, 20 had proven OB from which 15 had positive blood culture for Streptococcus pneumoniae. From these, 7 were fully or partially vaccinated. The serotype was identified on 14 cases: 6 were PCV10 vaccine serotypes (none of them vaccinated), 2 were serotype related (both partially vaccinated) and 6 were non vaccine serotypes (partially or totally vaccinated). Discussion: It is necessary to improve diagnostic methods for respiratory viruses and urinary tract infections and try to expand coverage of pneumococcal conjugated vaccines in the pediatric population in order to reduce the risk of invasive pneumococcal disease in Chile.

Introducción: La bacteriemia oculta (BO) es uno de los diagnósticos que se plantean en los niños bajo 3 años de edad que se presentan con fiebre sin foco en el servicio de urgencia. Objetivo: Describir el diagnóstico de BO luego de la introducción de la vacunación universal para Streptococcus pneumoniae en Chile. Materiales y Métodos: Revisión descriptiva de seguimiento prospectivo de datos de niños con diagnóstico de BO posible derivados del SU entre 2010 y 2013. Resultados: Se diagnosticó BO posible en 391 pacientes. En 233 pacientes se encontró foco, siendo infecciones respiratorias virales e infección urinaria las más frecuentes. En 158 pacientes se diagnosticó BO probable, en 20 BO probada y 15 tuvieron hemocultivos positivos para S. pneumoniae. De estos últimos 7 estaban total o parcialmente vacunados. Se identificó serotipo en 14 casos: 6 serotipos vaccinales incluidos en PCV10 (ninguno vacunado), 2 serotipos relacionados (ambos parcialmente protegido) y 6 serotipos no vaccinales (parcial o totalmente vacunados). Discusión: Es necesario mejorar las técnicas diagnósticas de infecciones respiratorias virales e infección urinaria e intentar ampliar la cobertura de las vacunas neumocóccicas conjugadas en la población pediátrica, para reducir el riesgo de enfermedades neumocóccicas invasoras en Chile.

Infección por VIH/SIDA en niños y adolescentes: cohorte chilena 1987-2014 / HIV/AIDS infection in children and adolescents: Chilean cohort 1987-2014

The present document describes the Cohort of HIV/AIDS children detected in Chile from 1987 to August 2014 and the effectiveness of the Protocol for Prevention of Vertical Transmission (PPVT) of HIV infection. Of the 375 HIV infected children enrolled since 1987 to August 2014, 245 of them are still in pediatric control. From the analysis of the Cohort is inferred that: a) it has observed an improvement in the detection of the HIV infected child, in number and precocious time; b) the majority of these children continue to be detected by clinic symptoms and signs (mainly unspecific and infectious manifestations); c) the ARVT use has meant a clinic and immunologic improvement with diminution of the infections, principally opportunistic infections, with a better life quality, a prolongation of survival and a diminution of lethality; d) as more survival has been produced, cancer has begun to be detected, a very infrequent complication observed in them before the ARVT use. The PPVT started in 1995, and was reinforced in 2005 with the "Joint Norm of HIV and Syphilis Vertical Transmission Prevention" (MINSAL), both have meant a diminution of the HIV vertical transmission from > 35% (before 1995) to < 2% nowadays in the mother-child binomial; also have permitted a second generation of HIV exposed children born without infection. In spite this PPVT, still HIV infected child continue to be detected which imply failures in some points of the health system.

Se presentan datos de la cohorte de niños con infección por VIH/SIDA detectados en Chile desde el año 1987 a agosto de 2014 y datos de la transmisión vertical (TV) del VIH con uso de protocolos de prevención de TV (PPTV). De los 375 niños infectados con VIH en este período, siguen en control pediátrico 245. Del análisis de la cohorte se desprende que: a) ha habido una mejoría en la pesquisa de los niños infectados con VIH, tanto en número como en precocidad; b) estos niños siguen detectándose, en su mayoría, por hechos clínicos (manifestaciones inespecíficas e infecciosas principalmente); c) el uso de TARV ha significado una mejoría clínica e inmunológica con disminución de las infecciones, principalmente las oportunistas, con una mejor calidad de vida, prolongación de la sobrevida, y disminución de la letalidad; d) por su mayor sobrevida, se ha observado el desarrollo de cánceres, muy infrecuentes en ellos antes del uso de terapia anti-retroviral. La aplicación de Protocolos de Prevención de la TV desde 1995, reforzada el 2005 con la “Norma Conjunta de la Prevención de la Transmisión Vertical del VIH y Sífilis” (MINSAL), ha significado una disminución de la TV del VIH desde más de 35% (antes de 1995) a < 2% actualmente en los binomios en prevención; además ha permitido que una segunda generación de niños expuestos al VIH nazca no infectada. A pesar de estos PPTV, aún siguen naciendo niños infectados con VIH, lo que implica fallas en algunos puntos del sistema de salud.

[HIV/AIDS infection in children and adolescents: Chilean cohort 1987-2014]. / Infección por VIH/SIDA en niños y adolescentes: cohorte chilena 1987-2014.

The present document describes the Cohort of HIV/AIDS children detected in Chile from 1987 to August 2014 and the effectiveness of the Protocol for Prevention of Vertical Transmission (PPVT) of HIV infection. Of the 375 HIV infected children enrolled since 1987 to August 2014, 245 of them are still in pediatric control. From the analysis of the Cohort is inferred that: a) it has observed an improvement in the detection of the HIV infected child, in number and precocious time; b) the majority of these children continue to be detected by clinic symptoms and signs (mainly unspecific and infectious manifestations); c) the ARVT use has meant a clinic and immunologic improvement with diminution of the infections, principally opportunistic infections, with a better life quality, a prolongation of survival and a diminution of lethality; d) as more survival has been produced, cancer has begun to be detected, a very infrequent complication observed in them before the ARVT use. The PPVT started in 1995, and was reinforced in 2005 with the "Joint Norm of HIV and Syphilis Vertical Transmission Prevention" (MINSAL), both have meant a diminution of the HIV vertical transmission from > 35% (before 1995) to < 2% nowadays in the mother-child binomial; also have permitted a second generation of HIV exposed children born without infection. In spite this PPVT, still HIV infected child continue to be detected which imply failures in some points of the health system.

Incidencia de cáncer en niños chilenos infectados por VIH / Incidence of cáncer in Chilean HIV-infected children

Background: Pediatric HIV (+) patients have a 100 times greater risk of cancer than HIV (-) children. Objective: To describe in Chilean HIV (+) children, cancer types, its appearance in relation to the stages of HIV disease and mortality. Methods: A protocol was created to know some characteristics of these patients from the point of view of their HIV infection and cancer pathology. Results: Of 360 HIV (+) children confirmed by the Institute of Public Health to May 2014, 9 patients with neoplastic disease (2.5%) were diagnosed. All the children were on ART, had more than three years of evolution of HIV infection and were in moderate to severe clinical/immunological stages. Lymphoma was the most common cancer. Five children, has received therapy according to Programa Infantil Nacional de Drogas Antineoplásicas (PINDA). There was no interaction between cancer treatment and antiretroviral therapy. Mortality was 13.8 x 1000 (5 cases). Conclusions: The incidence and type of neoplasia is consistent with the international literature, with less survival than HIV (+) children without tumors. The occurrence of cancer was observed in children with moderate to severe clinical and immunological compromise.

Introducción: Los pacientes pediátricos con infección por VIH tienen un riesgo 100 veces mayor de presentar cáncer que los niños no infectados. Objetivos: Describir en niños chilenos con infección por VIH, los tipos de cáncer, su aparición en relación a las etapas de la enfermedad por VIH y la letalidad. Material y Métodos: Se creó un protocolo para conocer algunas características de estos pacientes desde el punto de vista de su infección por VIH y su patología oncológica. Resultados: De 360 niños infectados confirmados por el Instituto de Salud Pública a mayo de 2014, se diagnosticaron nueve casos con patología oncológica (2,5%).Todos los niños estaban con TARV, tenían una evolución de infección por VIH mayor a 3 años, en etapas clínicas/inmunológicas moderada a grave. Linfoma fue el cáncer más frecuente. Cinco niños, recibieron terapia de acuerdo al Programa Infantil Nacional de Drogas Antineoplásicas (PINDA). No hubo interacción entre tratamiento anti-neoplásico y terapia anti-retroviral. La mortalidad fue de 13,8 x 1.000 (5 casos). Conclusiones: La incidencia y tipo de neoplasias está de acuerdo con lo comunicado en la literatura científica internacional, con sobrevida inferior a los niños con infección por VIH sin neoplasias. La aparición de cáncer se observó en niños con larga evolución y compromiso clínico e inmunológico moderado a grave.

[Incidence of cancer in Chilean HIV-infected children]. / Incidencia de cáncer en niños chilenos infectados por VIH.

BACKGROUND: Pediatric HIV (+) patients have a 100 times greater risk of cancer than HIV (-) children. OBJECTIVE: To describe in Chilean HIV (+) children, cancer types, its appearance in relation to the stages of HIV disease and mortality. METHODS: A protocol was created to know some characteristics of these patients from the point of view of their HIV infection and cancer pathology. RESULTS: Of 360 HIV (+) children confirmed by the Institute of Public Health to May 2014, 9 patients with neoplastic disease (2.5%) were diagnosed. All the children were on ART, had more than three years of evolution of HIV infection and were in moderate to severe clinical/immunological stages. Lymphoma was the most common cancer. Five children, has received therapy according to Programa Infantil Nacional de Drogas Antineoplásicas (PINDA). There was no interaction between cancer treatment and antiretroviral therapy. Mortality was 13.8 x 1000 (5 cases). CONCLUSIONS: The incidence and type of neoplasia is consistent with the international literature, with less survival than HIV (+) children without tumors. The occurrence of cancer was observed in children with moderate to severe clinical and immunological compromise.

[Sarcoptic mange: report of an outbreak in a family and their pet]. / Sarna sarcóptica: comunicación de un brote en un grupo familiar y su mascota.

Scabies caused by the genus Sarcoptes scabiei var canis is a prevalent infection in dogs and affects abandoned, malnourished and overcrowded animals, causing hair loss and an intensely pruritic crusting dermatitis. In humans the manifestation is a self-limiting pruritic dermatitis, but persistent cases are described. An outbreak of sarcoptic mange is reported in a family group (seven people, including a 5 month infant and his mother). The infective source was their own house dog who was taken from the street. The diagnosis was confirmed by the detection of mites and eggs in the acarotest of the dog and mites of S. scabei in the infant. Sarcoptic mange should be suspected in individuals with allergic dermatitis who have contact with dogs. Treatment in humans is usually symptomatic and may need miticides if the infection persists. The control of the disease requires an appropriate pet treatment.

[Fever of unknown origin: a challenge for the pediatric infectious diseases specialist]. / Síndrome febril prolongado: un desafío para el infectólogo pediatra.

INTRODUCTION: Prolonged febrile syndrome (PFS) is defined as fever 7-10 days, with initial study does not allow etiologic diagnosis. OBJECTIVE: To describe the main causes of the PFS and its temporal behavior in Pediatric Infectious Diseases Unit Outpatient Care of Complejo Asistencial Dr. Sótero del Río (CASR). PATIENTS AND METHODS: A descriptive, prospective study between january 2007-december 2012, about 153 patients from 6 weeks to 14 years 11 months old, diagnosed with PFS, tab completing clinical and laboratory monitoring. RESULTS: etiology was obtained in 67.9%, the causes were infection (88.4%), neoplasms (4.8%), rheumatological (4.8%) and Kawasaki disease (2.8%). The most important infectious causes were enteric fevers (typhoid and paratyphoid) (18.4%), urinary tract infection (11.9%), Bartonella henselae infections and adenovirus (8.7%) each one and Epstein Barr virus (7.6%). Ninety eight percent of patients had complete resolution, 60.7% did not require hospitalization and mortality was 0%. DISCUSSION: As in previous pediatric clinical series the infections were the most frequent causes. Enteric fever persists as principal cause, however, the epidemiological evidence is oscillating in time endorsing the local statistics can count over the years to improve the diagnostic and therapeutic approach.

Sarna sarcóptica: comunicación de un brote en un grupo familiar y su mascota / Sarcoptic mange: report of an outbreak in a family and their pet

Scabies caused by the genus Sarcoptes scabiei var canis is a prevalent infection in dogs and affects abandoned, malnourished and overcrowded animals, causing hair loss and an intensely pruritic crusting dermatitis. In humans the manifestation is a self-limiting pruritic dermatitis, but persistent cases are described. An outbreak of sarcoptic mange is reported in a family group (seven people, including a 5 month infant and his mother). The infective source was their own house dog who was taken from the street. The diagnosis was confirmed by the detection of mites and eggs in the acarotest of the dog and mites of S. scabei in the infant. Sarcoptic mange should be suspected in individuals with allergic dermatitis who have contact with dogs. Treatment in humans is usually symptomatic and may need miticides if the infection persists. The control of the disease requires an appropriate pet treatment.

La sarna producida por el género Sarcoptes scabiei var canis, infección prevalente en perros y de alto potencial zoonótico, afecta a animales abandonados, desnutridos y hacinados y causa alopecia y una dermatitis costrosa intensamente pruriginosa. En el ser humano produce una dermatitis pruriginosa generalmente autolimitada, pero se describen casos persistentes. Se reporta un brote de sarna sarcóptica en un grupo familiar (siete personas, incluidas una lactante y su madre) cuya fuente de infección fue su mascota canina recogida de la calle. El diagnóstico fue confirmado por visualización en el ácarotest de ácaros y huevos en el perro y ácaros de S. scabiei en la lactante. La sarna sarcóptica debe sospecharse en casos de dermatitis alérgica en personas con contacto con perros. El tratamiento en el humano, habitualmente sintomático, puede necesitar acaricidas si el cuadro persiste. El control de la enfermedad requiere el adecuado tratamiento de la mascota.

Síndrome febril prolongado: un desafío para el infectólogo pediatra / Fever of unknown origin: a challenge for the pediatric infectious diseases specialist

Introduction: Prolonged febrile syndrome (PFS) is defined as fever 7-10 days, with initial study does not allow etiologic diagnosis. Objective: To describe the main causes of the PFS and its temporal behavior in Pediatric Infectious Diseases Unit Outpatient Care of Complejo Asistencial Dr. Sótero del Río (CASR). Patients and Methods: A descriptive, prospective study between january 2007-december 2012, about 153 patients from 6 weeks to 14 years 11 months old, diagnosed with PFS, tab completing clinical and laboratory monitoring. Results: etiology was obtained in 67.9%, the causes were infection (88.4%), neoplasms (4.8%), rheumatological (4.8%) and Kawasaki disease (2.8%). The most important infectious causes were enteric fevers (typhoid and paratyphoid) (18.4%), urinary tract infection (11.9%), Bartonella henselae infections and adenovirus (8.7%) each one and Epstein Barr virus (7.6%). Ninety eight percent of patients had complete resolution, 60.7% did not require hospitalization and mortality was 0%. Discussion: As in previous pediatric clinical series the infections were the most frequent causes. Enteric fever persists as principal cause, however, the epidemiological evidence is oscillating in time endorsing the local statistics can count over the years to improve the diagnostic and therapeutic approach.

Introducción: El síndrome febril prolongado (SFP) se define como fiebre entre 7-10 días, con estudio inicial que no permite un diagnóstico etiológico. Objetivo: Describir las principales etiologías del SFP y su comportamiento temporal en la unidad de infectología pediátrica ambulatoria del Complejo Asistencial Dr. Sótero del Río (CASR). Pacientes y Método: Estudio descriptivo, prospectivo, entre enero de 2007-diciembre de 2012. Análisis de 153 pacientes entre 6 semanas y 14 años 11 meses de edad, con diagnóstico de SFP, que completaron ficha de seguimiento clínico-laboratorial. Resultados: Se obtuvo diagnóstico etiológico en 67,9%, las causas fueron: infecciones (88,4%), neoplasias (4,8%), reumatológicas (4,8%) y enfermedad de Kawasaki (2,8%). Las causas infecciosas más importantes fueron: fiebres entéricas (tifoidea y paratifoidea) (18,4%), infección del tracto urinario (11,9%), enfermedades por Bartonella henselae y adenovirus (8,7%) cada uno y virus de Epstein Barr (7,6%). El 98% de los pacientes tuvo resolución completa, 60,7% no requirió hospitalización y no se registraron decesos. Discusión: Como en las series clínicas antes publicadas, las infecciones fueron la causa más frecuente de SFP. La fiebre entérica persiste como causa principal; sin embargo, se evidencia una situación epidemiológica oscilante en el tiempo justificando la necesidad de contar con estadísticas locales a lo largo de los años para mejorar el enfoque diagnóstico y terapéutico.

Prevención de la transmisión vertical y efectos secundarios de la terapia anti-retroviral en hijos nacidos de madres seropositivas para VIH en el área Sur-Oriente de Santiago, Chile: perspectiva de 15 años / Prevention of vertical transmission and side effects of antiretroviral therapy in children born to HIV-positive South Eastern area of Santiago, Chile: 15-years perspective

Antiretroviral therapy (ART) has shown to be an effective measure in decreasing HIV vertical transmission (VT). Nevertheless, it is not free from adverse effects in the newborn: risk of prematurity, low birth weight, metabolic disorders, among others. Despite the importance of the subject, there are few national data that analyze the problem. We performed a retrospective analysis of a cohort of HIV positive mother/child binomial, followed between 1995 and 2010. Ninety-four pregnancies and 96 children (2 twin pregnancies) were analyzed. The rate of VT was 2.1%. Adverse effects attributed to ART were found on 85.4% of the newborn; highlighting the presence of anemia (70.8%) and several metabolic disorders [elevated lactate without acidosis (29.2%), lactic acidosis (12.5%), hyperkalemia (14.6%), metabolic acidosis (9.4%)]. Maternal exposure to protease inhibitors proved to be an independent risk factor for the development of metabolic disorders in newborns (OR 0.15 [0.04-0.48], p < 0.01). In our series, ART was effective in reducing the VT, however exposed newborns showed a high frequency of adverse effects, so it is advisable to implement programs for monitoring these patients to prevent sequelae.

La terapia anti-retroviral (TARV) es efectiva en disminuir la transmisión vertical (TV) del VIH, pero no está exenta de efectos adversos en los recién nacidos: riesgo de prematurez, bajo peso al nacer, alteraciones metabólicas y otros. Pese a lo relevante del tema, existen pocos datos nacionales que analicen el problema. Realizamos un estudio observacional, retrospectivo, de una serie de binomios madre infectada por VIH/hijo atendidos entre los años 1995 y 2010. Se analizaron 94 embarazos y 96 hijos (2 embarazos gemelares). La tasa de TV fue 2,1%. De los recién nacidos, 85,4% presentó efectos adversos atribuibles a la exposición a TARV destacando la presencia de anemia (70,8%) y alteraciones metabólicas varias [hiperlactacidemia sin acidosis (29,2%), acidosis láctica (12,5%), hiperkalemia (14,6%) y acidosis metabólica (9,4%). La exposición materna al uso de IP demostró ser un factor de riesgo independiente para el desarrollo de alteraciones metabólicas en los recién nacidos (OR 4 [1,58-10,12], p 0,003). En nuestra serie, la TARV demostró ser efectiva en reducir la TV. Sin embargo, los recién nacidos expuestos presentaron alta frecuencia de efectos adversos, por lo que es recomendable la implementación de programas de seguimiento de estos pacientes para prevenir secuelas.