Evaluation of a multiplex real-time PCR targeting the ß-tubulin gene for the detection and differentiation of Sporothrix schenckii and Sporothrix brasiliensis.

Sporothrix sp. is a thermally dimorphic genus of fungi known to cause subacute or chronic subcutaneous lesions in humans and animals and is the cause of increasing public health concern due to spread of feline-associated cases. Here, we adapted and evaluated a recently described real-time PCR assay targeting the ß-tubulin gene to rapidly detect and differentiate two related species, S. schenckii and S. brasiliensis. The assay was tested with 55 S. brasiliensis, 19 S. schenckii, and 85 isolates from other clinically relevant fungi, and showed 100% concordance with reference identification methods. The assay showed high analytical sensitivity with a limit of detection of 1 pg of DNA per microliter of sample for both targets. The assay was further evaluated with 11 fresh and 17 formalin-fixed, paraffin-embedded (FFPE) tissues. This multiplex real-time PCR assay successfully detected the DNA from both S. brasiliensis and S. schenckii isolates as well as S. schenckii from fresh and FFPE tissues. Our results demonstrate this assay performs well and could be a helpful molecular tool to support rapid species identification in cultures and primary specimens.IMPORTANCEHaving available molecular tools to identify and differentiate closely related species will allow clinical, veterinarians, and public health labs to provide diagnostic results with accuracy and short turnaround time for the routine and outbreak response activities.

Generation of a plasmid series for rapid sub-cloning and use in various Enterobacteriaceae.

Plasmids are molecular genetic tools used for trans-complementation and gene expression in bacteria. Challenges faced by researchers include limited repertoire of antibiotic resistance of plasmids, issues related to plasmid compatibility and restricted or incompatible multiple cloning sites when needing to change plasmid copy number to tune production of their protein of interest. In this study, a series of plasmids were generated with compatible multiple cloning sites and homologous DNA regions to allow for modular cloning for rapid exchange of antibiotic resistance and plasmid origin. Plasmids generated in this series have options for high, mid, and low plasmid copy number, and have either an integrated FLAG epitope in the multiple cloning site or possess an uninterrupted multiple cloning site with the option of using the common LacZ-based blue/white screening method. Low copy plasmids also have one of five antibiotic selection markers. To demonstrate functionality of these plasmids, a representative FLAG tagged protein and mCherry were cloned into the low copy plasmids and expressed in various bacteria belonging to the Enterobacteriaceae family. In conclusion, by creating a new plasmid series, we have expanded the toolkit of available molecular biology tools for bacterial work.

Strategies of Pharmacological Repositioning for the Treatment of Medically Relevant Mycoses.

Fungal infections represent a worldwide concern for public health, due to their prevalence and significant increase in cases each year. Among the most frequent mycoses are those caused by members of the genera Candida, Cryptococcus, Aspergillus, Histoplasma, Pneumocystis, Mucor, and Sporothrix, which have been treated for years with conventional antifungal drugs, such as flucytosine, azoles, polyenes, and echinocandins. However, these microorganisms have acquired the ability to evade the mechanisms of action of these drugs, thus hindering their treatment. Among the most common evasion mechanisms are alterations in sterol biosynthesis, modifications of drug transport through the cell wall and membrane, alterations of drug targets, phenotypic plasticity, horizontal gene transfer, and chromosomal aneuploidies. Taking into account these problems, some research groups have sought new therapeutic alternatives based on drug repositioning. Through repositioning, it is possible to use existing pharmacological compounds for which their mechanism of action is already established for other diseases, and thus exploit their potential antifungal activity. The advantage offered by these drugs is that they may be less prone to resistance. In this article, a comprehensive review was carried out to highlight the most relevant repositioning drugs to treat fungal infections. These include antibiotics, antivirals, anthelmintics, statins, and anti-inflammatory drugs.

Subcutaneous phaeohyphomycosis due to Phaeoacremonium venezuelense: The first clinical case report in Costa Rica.

Phaeoacremonium is a genus of dematiaceous fungi that rarely causes human infections. We describe a case of subcutaneous infection in a 70-year-old diabetic man with lesions on the dorsum of the one foot. The agent was isolated, and for the final identification we performed matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and DNA sequencing. After diagnosis, the patient underwent curettage of the cyst and received 100mg of Itraconazole, twice daily for 6 months. Clinical resolution of the lesion was observed after treatment. This is the first case of infection by Phaeoacremonium venezuelense reported in Costa Rica.

Physical exercise and major depressive disorder in adults: systematic review and meta-analysis.

The objective of this study was to assess the benefits and potential risks associated with different physical exercise modalities for managing symptoms in adults with major depressive disorder who were not receiving second-generation antidepressants or cognitive behavioral therapy. A systematic review and meta-analysis of randomized controlled trials (RCTs) were conducted. The search included multiple databases: Medline, Cochrane Central Register of Controlled Trials (CENTRAL), Embase, PsycInfo, Web of Science, Clinical Trials repository, gray literature, and manual search. No language restrictions were applied. Eligible studies involved RCTs of adults with major depressive disorder who were not on antidepressants or receiving psychological therapy, comparing various exercise modalities with second-generation antidepressants or cognitive behavioral therapy, body-mind exercise, or no exercise interventions. Nine RCTs involving 678 adults were analyzed. The pooled results indicated a small clinical effect favoring exercise in reducing depressive symptoms, although the difference was not statistically significant (SMD = 0.27, 95% CI [- 0.58, 0.04], P = 0.09). Subgroup analyses suggested that intervention duration, frequency, intensity, supervision, age, overweight/obesity status, and diagnosis of depression could influence treatment outcomes. A sensitivity analysis was conducted for studies with controls without exercise interventions and a low risk of bias in the domains related to the randomization process and deviations from the intended interventions. The results showed that there are no statistically significant differences when interventions are compared with medication and body-mind exercise (p = 0.12, I2 = 78%). Furthermore, the analysis showed a moderate effect size favoring exercise, but no statistically significant difference between groups (p = 0.05), with high heterogeneity (I2 = 85%). The evidence quality was generally low to very low, and methodological limitations compromised the certainty of the findings. Adverse events associated with exercise were manageable. The study emphasizes the need for well-designed RCTs to provide clearer insights into the potential benefits of exercise in managing major depressive disorder symptoms. Caution is warranted in interpreting these results due to the limitations of the included studies.Systematic review registration: PROSPERO CRD42022356741.

Application of Real-Time PCR Assays for the Diagnosis of Histoplasmosis in Human FFPE Tissues Using Three Molecular Targets.

Histoplasmosis is a fungal infection caused by the thermally dimorphic fungus Histoplasma capsulatum. This infection causes significant morbidity and mortality in people living with HIV/AIDS, especially in countries with limited resources. Currently used diagnostic tests rely on culture and serology but with some limitations. No molecular assays are commercially available and the results from different reports have been variable. We aimed to evaluate quantitative real-time PCR (qPCR) targeting three protein-coding genes of Histoplasma capsulatum (100-kDa, H and M antigens) for detection of this fungus in formalin-fixed paraffin-embedded (FFPE) samples from patients with proven histoplasmosis. The sensitivity of 100-kDa, H and M qPCR assays were 93.9%, 91% and 57%, respectively. The specificity of 100-kDa qPCR was 93% when compared against samples from patients with other mycoses and other infections, and 100% when samples from patients with non-infectious diseases were used as controls. Our findings demonstrate that real-time PCR assays targeting 100-kDa and H antigen showed the most reliable results and can be successfully used for diagnosing this mycosis when testing FFPE samples.

Conocimiento del parto humanizado en Colombia en residentes y ginecólogos / Knowledge of humanized childbirth in Colombian residents and gynecologists

Resumen OBJETIVO: Identificar, mediante una encuesta, el conocimiento de especialistas y residentes de Ginecología y Obstetricia de los beneficios de la atención respetuosa y digna a la paciente en el momento del parto. MATERIALES Y MÉTODOS: Estudio observacional, descriptivo, transversal y prospectivo efectuado en una muestra de ginecoobstetras y residentes de diferentes universidades e instituciones de Colombia a quienes se aplicó, entre los meses de diciembre de 2021 a agosto de 2022, un instrumento estructurado de manera individual, presencial o virtual. RESULTADOS: Se obtuvieron 343 respuestas. El 51% de la muestra fue de especialistas con más de dos años de experiencia. El 37.2% de los residentes cursaba el tercer y 20.2% el segundo año. Más de la mitad de los encuestados no recibió ni recibe educación del tema. Se identificó un vacío en el conocimiento y su implementación. La totalidad de la muestra no reconoció algún beneficio materno o neonatal. El 86.3% consideró que no existe contraindicación para la implementación del parto digno y respetuoso y el 94.8% manifestó una correlación entre la desinformación y su baja implementación. El 69.4% de la muestra conocía y ponía en práctica el plan de parto en su práctica diaria. CONCLUSIONES: De la muestra analizada se concluye que hay desinformación acerca de los beneficios maternos y neonatales del parto digno y respetuoso por parte de residentes y especialistas de Ginecología y Obstetricia. Por lo tanto, es necesario aumentar los contenidos del tema a los programas de educación. Además, estandarizar los protocolos ajustados que faciliten su ejecución e implementación.

Abstract OBJECTIVE: To identify, by means of a survey, the knowledge of specialists and residents in Gynaecology and Obstetrics of the benefits of respectful and dignified care for the patient at the time of delivery. MATERIALSAND METHODS: Observational, descriptive, cross-sectional, and prospective study carried out on a sample of obstetrician-gynecologists and residents from different universities and institutions in Colombia to whom a structured instrument was applied individually, in person or online, between December 2021 and August 2022. RESULTS: 343 responses were obtained. Fifty-one percent of the sample were specialists with more than two years of experience. 37.2% of the residents were in their third year and 20.2% were in their second year. More than half of the respondents did not and do not receive any education on the subject. A gap in knowledge and implementation were identified. The entire sample did not recognize any maternal or neonatal benefit. 86.3% considered that there is no contraindication to the implementation of respectful and dignified childbirth and 94.8% expressed a correlation between misinformation and low implementation of respectful and dignified childbirth. 69.4% of the sample were aware of and implemented the birth plan in their daily practice. CONCLUSIONS: From the sample analyzed, it is concluded that there is misinformation about the maternal and neonatal benefits of dignified and respectful childbirth on the part of Gynaecology and Obstetrics residents and specialists. Therefore, it is necessary to increase the content of the subject in education programs. In addition, it is necessary to standardize adjusted protocols that facilitate their execution and implementation.

Pólipo endometrial metaplásico: hallazgo histeroscópico de enfermedad premaligna en mujeres jóvenes. Reporte de tres casos / Endometrial metaplastic polyps: findings of premalignant disease on histeroscopy in young women, a report of 3 cases

ANTECEDENTES: Los pólipos endometriales son sobrecrecimientos de la mucosa endometrial, una de las causas más comunes de hemorragia uterina anormal, pueden estar asociados con lesiones premalignas y malignas del endometrio. Existen características histeroscópicas que ayudan a diferenciar una lesión benigna o maligna. El pólipo metaplásico es un hallazgo histeroscópico caracterizado por superficies laminares e hiperqueratosis, con zonas blancas e hiperrefringentes. CASOS CLÍNICOS: Tres casos clínicos de pacientes menores de 40 años, con hemorragia uterina anormal donde el hallazgo histeroscópico común fue el pólipo metaplásico, hallazgo premaligno que se confirmó mediante el análisis patológico y la positividad del marcador de inmunohistoquímica p63. Caso 1. Paciente de 28 años, nulípara, obesa, con antecedente de síndrome de ovario poliquístico, con engrosamiento endometrial e histeroscopia, con pólipo con superficie hiperqueratósica, a manera de láminas irregulares, con excrecencias y lesiones exofíticas. Caso 2. Paciente de 25 años, con endometrio irregular engrosado e histeroscopia con hallazgo de pólipos transparentes, con cambios vasculares leves y superficie con zonas hiperrefringentes blanquecinas. Caso 3. Paciente de 38 años, con antecedente de síndrome de ovario poliquístico y obesidad. En la histeroscopia el endometrio se encontró hiperplásico, irregular, con cambios vasculares y pólipos con zonas superficiales hiperrefringentes, sólidas y blancas. CONCLUSIONES: En pacientes jóvenes, con hallazgo histeroscópico de pólipo metaplásico debe considerarse y descartar la enfermedad premaligna y maligna endometrial. El estudio debe completarse con marcadores de inmunohistoquímica (p63), específicos de la metaplasia escamosa.

Abstract BACKGROUND: Endometrial polyps are overgrowths of the endometrial mucosa, one of the most common causes of abnormal uterine bleeding and may be associated with premalignant and malignant endometrial lesions. There are hysteroscopic features that help differentiate a benign or malignant lesion. Metaplastic polyp is a hysteroscopic finding characterized by lamellar surfaces and hyperkeratosis, with white and hyperreflective areas. CLINICAL CASES: Three clinical cases of patients under 40 years of age, with abnormal uterine bleeding where the common hysteroscopic finding was metaplastic polyp, a premalignant finding that was confirmed by pathological analysis and positivity of the immunohistochemistry marker p63. Case 1. 28-year-old female, nulliparous, obese, with a history of polycystic ovary syndrome, with endometrial thickening and hysteroscopy, with polyp with hyperkeratotic surface, in the form of irregular sheets, with excrescences and exophytic lesions. Case 2. 25-year-old patient, with irregular thickened endometrium and hysteroscopy with finding of transparent polyps, with mild vascular changes and surface with whitish hyperrefringent areas. Case 3. 38-year-old patient with a history of polycystic ovary syndrome and obesity. At hysteroscopy the endometrium was found to be irregularly hyperplastic, with vascular changes and polyps with hyper-refringent, solid, white superficial areas. CONCLUSIONS: In young patients, with hysteroscopic finding of metaplastic polyp should be considered and premalignant and malignant endometrial disease should be ruled out. The study should be completed with immunohistochemical markers (p63), specific for squamous metaplasia.

Emergency severity level-3 patient flow based on point-of-care testing improves patient outcomes.

BACKGROUND: Overcrowding of the Emergency Department is rapidly becoming a global challenge and a major source of concern for emergency physicians. The desire to improve Emergency Department throughput requires novel approaches to patient flow. MATERIALS AND METHODS: We conducted a prospective and cluster-randomized study, to evaluate the impact in patient outcomes of a new patient flow based on Point-of-Care Testing (POCT). A total of 380 Emergency Severity Level-3 patients were enrolled and studied in two different groups, interventional arm (laboratory analyses performed on POCT analyzers implemented in the Emergency Department) or control arm (central laboratory). The primary outcome was the Emergency Department length of stay. Secondary outcome included the time to first medical intervention, the laboratory turnaround time and the time to disposition decision. Readmission within the 7 days after discharge was also calculated. RESULTS: Length of stay significantly decreased by 88.50 min (from 247.00 to 158.50), time to disposition decision by 89.00 min (from 192.00 to 103.00) and laboratory turnaround time by 67.11 min (from 89.84 to 22.73) in the POCT group. No increase in readmission was found. CONCLUSION: Our strategy based on POCT represents a good approach to optimize patient flow in the Emergency Department and it should be seen as a starting point for further studies focusing on improving throughput.

Methylphenidate ameliorates the homeostatic balance between levels of kynurenines in ADHD children.

The kynurenine pathway of tryptophan metabolism has been involved in ADHD We quantified basal levels and daily fluctuations of tryptophan and several kynurenine metabolites, as well as their changes after treatment with methylphenidate (MPH). A total of 179 children were recruited, grouped into ADHD (n = 130) and healthy controls (CG,n = 49). Blood samples were drawn at 20:00 and 09:00 h and only in the ADHD group after 4.63±2.3 months of treatment. Nocturnal urine was collected between both draws. Factorial analysis (Stata12.0) was performed with Groups, Time, Hour of Day and Depressive Symptoms (DS) as factors. MPH significantly increased plasma Kynurenic acid (2.4 ± 1.03/2.78±1.3 ng/mL; baseline/post-treatment, morning; z = 1.96,p<0.05) and Xanthurenic acid (2.39±0.95/2.88±1.19 ng/mL; baseline/post, morning; z = 2.7,p<0.007) levels, both with higher values in the evening. In DS+ patients, MPH caused a pronounced decrease in evening Anthranilic acid [3.08±5.02/ 1.82±1.46 ng/mL, z = 2.68,p = 0.0074] until matching values to other subgroups. In urine, MPH decreased the excretion of both Nicotinamide and Quinolinic acids, but only in the DS- subgroup. The kynurenine pathway may participate in the highly clinical favorable response to MPH. The observed changes could be considered as protective (i.e. increased plasma kynurenic acid vs. decreased quinolinic acid excretion) based on the knowledge of its physiological homeostatic functions.

Indole Tryptophan Metabolism and Cytokine S100B in Children with Attention-Deficit/Hyperactivity Disorder: Daily Fluctuations, Responses to Methylphenidate, and Interrelationship with Depressive Symptomatology.

Background: Indole tryptophan metabolites (ITMs), mainly produced at the gastrointestinal level, participate in bidirectional gut-brain communication and have been implicated in neuropsychiatric pathologies, including attention-deficit/hyperactivity disorder (ADHD). Method: A total of 179 children, 5-14 years of age, including a healthy control group (CG, n = 49), and 107 patients with ADHD participated in the study. The ADHD group was further subdivided into predominantly attention deficit (PAD) and predominantly hyperactive impulsive (PHI) subgroups. Blood samples were drawn at 20:00 and 09:00 hours, and urine was collected between blood draws, at baseline and after 4.63 ± 2.3 months of methylphenidate treatment in the ADHD group. Levels and daily fluctuations of ITM were measured by tandem mass spectrometer, and S100B (as a glial inflammatory marker) by enzyme-linked immunosorbent assay. Factorial analysis of variance (Stata 12.0) was performed with groups/subgroups, time (baseline/after treatment), hour of day (morning/evening), and presence of depressive symptoms (DS; no/yes) as factors. Results: Tryptamine and indoleacetic acid (IAA) showed no differences between the CG and ADHD groups. Tryptamine exhibited higher evening values (p < 0.0001) in both groups. No changes were associated with methylphenidate or DS. At baseline, in comparison with the rest of study sample, PHI with DS+ group showed among them much greater morning than evening IAA (p < 0.0001), with treatment causing a 50% decrease (p = 0.002). Concerning indolepropionic acid (IPA) MPH was associated with a morning IPA decrease and restored the daily profile observed in the CG. S100B protein showed greater morning than evening concentrations (p = 0.001) in both groups. Conclusion: Variations in ITM may reflect changes associated with the presence of DS, including improvement, among ADHD patients.

Embarazo ectópico cervical / Cervical Ectopic Pregnancy

El embarazo ectópico cervical representa alrededor de 0,1 por ciento de todos los embarazos ectópicos. La incidencia se estima en 1: 2500 a 1: 98,000 embarazos. Es un fenómeno extraño; sin embargo, la morbilidad y la mortalidad por esta causa son elevadas, pero pueden disminuirse con el diagnóstico temprano. Es extremadamente peligroso porque el trofoblasto puede alcanzar los vasos uterinos a través de la delgada pared cervical y provocar una hemorragia grave e incoercible que aun en la actualidad suele terminar en cirugías mutilantes que limitarán la reproducción futura. En la actualidad, no existen criterios claros en la bibliografía que ayuden a la toma de decisiones. Se reporta un caso de embarazo cervical tratado exitosamente mediante histerectomía total abdominal y posterior seguimiento en la Unidad de Cuidados Intensivos. Se exponen algunos criterios que en el futuro podrían ayudar a diagnosticar esta entidad devastadora de forma oportuna para disminuir la morbimortalidad por esta causa y mejorar las posteriores posibilidades reproductivas de la mujer(AU)

Cervical ectopic pregnancy accounts for about 0.1 percent of all ectopic pregnancies. The incidence is estimated at 1: 2500 to 1: 98,000 pregnancies. It is a strange phenomenon; however, morbidity and mortality from this cause are high, but early diagnosis can help reducing. It is extremely dangerous because the trophoblast can reach the uterine vessels through the thin cervical wall and cause a severe and incoercible hemorrhage that, even today, usually ends in mutilating surgeries that will limit future reproduction. At present, no clear criteria in the literature help decision-making. We report a case of cervical pregnancy successfully treated by total abdominal hysterectomy and subsequent follow-up in the Intensive Care Unit. We present some criteria that could help timely diagnosing this devastating entity to reduce morbidity and mortality and improve the later reproductive possibilities of women(AU)

Standardization and validation of real time PCR assays for the diagnosis of histoplasmosis using three molecular targets in an animal model.

Histoplasmosis is considered one of the most important endemic and systemic mycoses worldwide. Until now few molecular techniques have been developed for its diagnosis. The aim of this study was to develop and evaluate three real time PCR (qPCR) protocols for different protein-coding genes (100-kDa, H and M antigens) using an animal model. Fresh and formalin-fixed and paraffin-embedded (FFPE) lung tissues from BALB/c mice inoculated i.n. with 2.5x106 Histoplasma capsulatum yeast or PBS were obtained at 1, 2, 3, 4, 8, 12 and 16 weeks post-infection. A collection of DNA from cultures representing different clades of H. capsulatum (30 strains) and other medically relevant pathogens (36 strains of related fungi and Mycobacterium tuberculosis) were used to analyze sensitivity and specificity. Analytical sensitivity and specificity were 100% when DNAs from the different strains were tested. The highest fungal burden occurred at first week post-infection and complete fungal clearance was observed after the third week; similar results were obtained when the presence of H. capsulatum yeast cells was demonstrated in histopathological analysis. In the first week post-infection, all fresh and FFPE lung tissues from H. capsulatum-infected animals were positive for the qPCR protocols tested except for the M antigen protocol, which gave variable results when fresh lung tissue samples were analyzed. In the second week, all qPCR protocols showed variable results for both fresh and FFPE tissues. Samples from the infected mice at the remaining times post-infection and uninfected mice (controls) were negative for all protocols. Good agreement was observed between CFUs, histopathological analysis and qPCR results for the 100-kDa and H antigen protocols. We successfully standardized and validated three qPCR assays for detecting H. capsulatum DNA in fresh and FFPE tissues, and conclude that the 100-kDa and H antigen molecular assays are promising tests for diagnosing this mycosis.

Time-to-Boundary Function to Study the Development of Upright Stance Control in Children.

BACKGROUND: The development of postural control across the primary school time horizon is a complex process, which entails biomechanics modifications, the maturation of cognitive ability and sensorimotor organization, and the emergence of anticipatory behaviour. Postural stability in upright stance has been thus object of a multiplicity of studies to better characterize postural control in this age span, with a variety of methodological approaches. The analysis of the Time-to-Boundary function (TtB), which specifies the spatiotemporal proximity of the Centre of Pressure (CoP) to the stability boundaries in the regulation of posture in upright stance, is among the techniques used to better characterize postural stability in adults, but, as of now, it has not yet been introduced in developmental studies. The aim of this study was thus to apply this technique to evaluate the development of postural control in a sample population of primary school children. METHODS: In this cross-sectional study, upright stance trials under eyes open and eyes closed were administered to 107 healthy children, divided into three age groups (41 for Seven Years' Group, Y7; 38 for Nine Years' Group, Y9; 28 for Eleven Years' Group, Y11). CoP data were recorded to calculate the Time-to-Boundary function (TtB), from which four spatio-temporal parameters were extracted: the mean value and the standard deviation of TtB minima (Mmin, Stdmin), and the mean value and the standard deviation of the temporal distance between two successive minima (Mdist, Stddist). RESULTS: With eyes closed, Mmin and Stdmin significantly decreased and Mdist and Stddist increased for the Y7 group, at Y9 Mmin significantly decreased and Stddist increased, while no effect of vision resulted for Y11. Regarding age groups, Mmin was significantly higher for Y9 than Y7, and Stdmin for Y9 was higher than both Y7 and Y11; Mdist and Stddist resulted higher for Y11 than for Y9. CONCLUSION: From the combined results from the spatio-temporal TtB parameters, it is suggested that, at 9 years, children look more efficient in terms of exploring their limits of stability than at 7, and at 11 the observed TtB behaviour hints at the possibility that, at that age, they have almost completed the maturation of postural control in upright stance, also in terms of integration of the spatio-temporal information.

BDNF concentrations and daily fluctuations differ among ADHD children and respond differently to methylphenidate with no relationship with depressive symptomatology.

RATIONALE: Brain-derived neurotrophic factor (BDNF) enhances the growth and maintenance of several monoamine neuronal systems, serves as a neurotransmitter modulator and participates in the mechanisms of neuronal plasticity. Therefore, BDNF is a good candidate for interventions in the pathogenesis and/or treatment response of attention deficit hyperactivity disorder (ADHD). OBJECTIVE: We quantified the basal concentration and daily fluctuation of serum BDNF, as well as changes after methylphenidate treatment. METHOD: A total of 148 children, 4-5 years old, were classified into groups as follows: ADHD group (n = 107, DSM-IV-TR criteria) and a control group (CG, n = 41). Blood samples were drawn at 2000 and 0900 hours from both groups, and after 4.63 ± 2.3 months of treatment, blood was drawn only from the ADHD group for BDNF measurements. Factorial analysis was performed (Stata software, version 12.0). RESULTS: Morning BDNF (36.36 ± 11.62 ng/ml) in the CG was very similar to that in the predominantly inattentive children (PAD), although the evening concentration in the CG was higher (CG 31.78 ± 11.92 vs PAD 26.41 ± 11.55 ng/ml). The hyperactive-impulsive group, including patients with comorbid conduct disorder (PHI/CD), had lower concentrations. Methylphenidate (MPH) did not modify the concentration or the absence of daily BDNF fluctuations in the PHI/CD children; however, MPH induced a significant decrease in BDNF in PAD and basal day/night fluctuations disappeared in this ADHD subtype. This profile was not altered by the presence of depressive symptoms. CONCLUSIONS: Our data support a reduction in BDNF in untreated ADHD due to the lower concentrations in PHI/CD children, which is similar to other psychopathologic and cognitive disorders. MPH decreased BDNF only in the PAD group, which might indicate that BDNF is not directly implicated in the methylphenidate-induced amelioration of the neuropsychological and organic immaturity of ADHD patients.

Childhood histoplasmosis in Colombia: Clinical and laboratory observations of 45 patients.

Histoplasmosis is an important mycosis in the Americas; and in children with no immune system abnormalities, histoplasmosis is typically a self-limited process. In contrast, in children with immune problems, disease manifestations are frequently more severe and include dissemination. From 1984 to 2010, a retrospective study of paediatric patients who had been diagnosed with histoplasmosis was performed. A total of 45 pediatric cases of histoplasmosis were identified. The most important risk factor was malnutrition (37%), followed by environmental exposure (33%). The patients exhibited pulmonary infiltrates (83%), fever (76%), cough, constitutional symptoms (38%), headache (35%), and lymph node hypertrophy (33%). Concerning the clinical forms, 64% of the patients presented with the progressive disseminated form that frequently affected the central nervous system (48%). Diagnostic laboratory tests indicated that the cultures were positive for 80% of the patients, the agar gel immunodiffusion was reactive in 95%, the M band of the precipitate was more commonly observed (81%), and the complement fixation tests were reactive in 88% of the patients. The timely diagnosis of histoplasmosis is important, and for this reason, it is hoped that the results of this study will lead pediatricians toward a better understanding of this mycosis in children.

Whole-Genome Sequencing to Determine Origin of Multinational Outbreak of Sarocladium kiliense Bloodstream Infections.

We used whole-genome sequence typing (WGST) to investigate an outbreak of Sarocladium kiliense bloodstream infections (BSI) associated with receipt of contaminated antinausea medication among oncology patients in Colombia and Chile during 2013-2014. Twenty-five outbreak isolates (18 from patients and 7 from medication vials) and 11 control isolates unrelated to this outbreak were subjected to WGST to elucidate a source of infection. All outbreak isolates were nearly indistinguishable (<5 single-nucleotide polymorphisms), and >21,000 single-nucleotide polymorphisms were identified from unrelated control isolates, suggesting a point source for this outbreak. S. kiliense has been previously implicated in healthcare-related infections; however, the lack of available typing methods has precluded the ability to substantiate point sources. WGST for outbreak investigation caused by eukaryotic pathogens without reference genomes or existing genotyping methods enables accurate source identification to guide implementation of appropriate control and prevention measures.

Characterising atypical Candida albicans clinical isolates from six third-level hospitals in Bogotá, Colombia.

BACKGROUND: Candida species are the most frequently found fungal pathogens causing nosocomial disease in a hospital setting. Such species must be correctly identified to ensure that appropriate control measures are taken and that suitable treatment is given for each species. Candida albicans is causing most fungal disease burden worldwide; the challenge lies in differentiating it from emerging atypical, minor and related species such as Candida dubliniensis and Candida africana. The purpose of this study was to compare identification based on MALDI-TOF MS to standard identification systems using a set of nosocomial isolates. METHODS: Eleven nosocomial samples were collected from 6 third-level hospitals in Bogotá, Colombia. All the samples were identified by combining MALDI-TOF MS with morphological characters, carbohydrate assimilation and molecular markers (D1/D2 and HWP1). RESULTS: The present work describes the first collection of atypical Colombian Candida clinical isolates; these were identified as Candida albicans/Candida africana by their MALDI-TOF MS profile. Phenotypical characteristics showed that they were unable to produce chlamydospores, assimilate trehalose, glucosamine, N- acetyl-glucosamine and barely grew at 42 °C, as would be expected for Candida africana. The molecular identification of the D1/D2 region of large subunit ribosomal RNA and HWP1 hyphal cell wall protein 1 sequences from these isolates was consistent with those for Candida albicans. The mass spectra obtained by MALDI-TOF MS were analysed by multi-dimensional scaling (MDS) and cluster analysis, differences being revealed between Candida albicans, Candida africana, Candida dubliniensis reference spectra and two clinical isolate groups which clustered according to the clinical setting, one of them being clearly related to C. albicans. CONCLUSION: This study highlights the importance of using MALDI-TOF MS in combination with morphology, substrate assimilation and molecular markers for characterising Candida albicans-related and atypical C. albicans species, thereby overcoming conventional identification methods. This is the first report of hospital-obtained isolates of this type in Colombia; the approach followed might be useful for gathering knowledge regarding local epidemiology which could, in turn, have an impact on clinical management. The findings highlight the complexity of distinguishing between typical and atypical Candida albicans isolates in hospitals.

Music Training Increases Phonological Awareness and Reading Skills in Developmental Dyslexia: A Randomized Control Trial.

There is some evidence for a role of music training in boosting phonological awareness, word segmentation, working memory, as well as reading abilities in children with typical development. Poor performance in tasks requiring temporal processing, rhythm perception and sensorimotor synchronization seems to be a crucial factor underlying dyslexia in children. Interestingly, children with dyslexia show deficits in temporal processing, both in language and in music. Within this framework, we test the hypothesis that music training, by improving temporal processing and rhythm abilities, improves phonological awareness and reading skills in children with dyslexia. The study is a prospective, multicenter, open randomized controlled trial, consisting of test, rehabilitation and re-test (ID NCT02316873). After rehabilitation, the music group (N = 24) performed better than the control group (N = 22) in tasks assessing rhythmic abilities, phonological awareness and reading skills. This is the first randomized control trial testing the effect of music training in enhancing phonological and reading abilities in children with dyslexia. The findings show that music training can modify reading and phonological abilities even when these skills are severely impaired. Through the enhancement of temporal processing and rhythmic skills, music might become an important tool in both remediation and early intervention programs.Trial Registration: ClinicalTrials.gov NCT02316873

Prevalencia de vaginosis bacteriana, actinomyces spp, candida spp. y trichomonas vaginalis en usuarias del dispositivo intrauterino (DIU), Medellín, Colombia, 2011-2013 / Prevalence of bacterial vaginosis, actinomyces spp, candida spp, and trichomonas vaginalis in intrauterine device (IUD) users, Medellin, Colombia, 2011-2013

Introducción: el uso del dispositivo intrauterino incrementa el riesgo de algunas infecciones genitales. Objetivo: determinar la prevalencia de vaginosis bacteriana, Actinomyces spp., Candida spp. y Trichomonasvaginalis en usuarias del dispositivo intrauterino. Materiales y métodos: se realizó un estudio transversal en usuarias del dispositivo intrauterino, atendidas en un programa de tamización de cáncercérvico-uterino en Medellín, Colombia, entre 2011 y 2013. Se empleó una fuente de información secundaria basada en los registros citológicos del laboratorio clínico, se calculó la prevalencia global de las cuatro infecciones y las prevalencias específicas según grupo etario y sector de residencia, con un intervalo de confianza del 95%. La exploración de asociaciones se hizo con la prueba chi cuadrado de Pearson. Resultados: Se registraron 12.541 usuarias del dispositivo intrauterino de 10 comunas de Medellín. La edad promedio fue 34,0±9,8 años; el 50% de los valores centrales estuvo entre 26 y 41 años. Las prevalencias de infecciones vaginales fueron: vaginosis bacteriana 25,6%, Actinomyces spp. 8,9%, Candida spp. 5,1% y Trichomonas vaginalis 1,2%. La prevalencia de vaginosisbacteriana y Candida spp. fue estadísticamente mayor en adolescentes y jóvenes. La prevalencia de las cuatro infecciones fue estadísticamente diferente según el sector de residencia. Conclusión: en usuarias del dispositivo intrauterino las principales infecciones genitales son vaginosis bacteriana y Actinomyces spp., las adolescentes y jóvenes son los grupos de mayor riesgo para vaginosis bacterianay Candida spp. y la ocurrencia de infecciones varía entre los sectores de la ciudad; información relevante para la planeación de programas de prevención y atención.

Introduction: the use of intrauterine device increases the risk of genital infections. Objective: To determine the prevalence of bacterial vaginosis, Actinomyces spp., Candida spp., and Trichomonas vaginalis in intrauterine device users. Material and methods: A cross-sectional study was performed in intrauterine device users attended in a screening program for cervical cancer in Medellín, Colombia, between 2011 and 2013. A source of information was secondary, based on the records of the clinical laboratory of cytology. The overall prevalence of the four infections and specific prevalence by age group and residence area was calculated, all with confidence intervals of 95%. To explore associationsPearson chi-square test was used. Results: It was registered 12,541 users of the intrauterine device from 10 districts of Medellin. The users mean age was 34.0±9.8 years; 50% of the central values was between 26 and 41 years. The prevalence of vaginal infections were: bacterial vaginosis 25.6%, Actinomyces spp. 8.9%, Candida spp. 5.1% and Trichomonas vaginalis 1.2%. The prevalence of bacterial vaginosis and Candida spp. was statistically higher in adolescents and youth. The prevalenceof the four infections was statistically different according to the residence sector.Conclusion: in intrauterine devices users the major genital infections are bacterial vaginosis and Actinomycesspp. Adolescents and young people are the groups most at risk for bacterial vaginosis and Candida spp. and the occurrence of infections varies between sectors of the city; information that is relevant for planning prevention and care programs.