Comparison between Two Molecular Techniques: Nested and Real-Time Polymerase Chain Reaction Targeting 100-kDa Hc Protein for Detection of Histoplasma capsulatum in Environmental Samples.

Histoplasmosis, one of the most frequent endemic mycoses in the Americas, is caused by the inhalation of airborne conidia of Histoplasma capsulatum. Better understanding of the distribution of this fungus in the environment is important for the development of appropriate public health measures to prevent human infections. Previously, we used Hc100 nested polymerase chain reaction (PCR) to identify H. capsulatum DNA in 10% of environmental samples in Colombia. Here, we validate a 100-kDa real-time PCR assay for the detection of this fungus in the environment. Using this method, we identified H. capsulatum DNA in 80% of samples of raw organic materials, such as chicken manure, soil from caves, and bird and bat guano, as well as in 62% of samples of organic fertilizer that underwent the composting process. We demonstrated that 100-KDa real-time PCR is a useful tool for environmental surveillance that can be used to identify the potential reservoirs of H. capsulatum and to prevent outbreaks, especially in people with the higher risk of exposure, such as spelunkers, farmers, poultry manure collectors, and anyone who handle organic fertilizers or bat and bird excreta.

Molecular epidemiology of Colombian Histoplasma capsulatum isolates obtained from human and chicken manure samples.

The thermally dimorphic fungus Histoplasma capsulatum is the causative agent of histoplasmosis, one of the most prevalent endemic mycosis in the Americas. In tropical regions, agro-ecosystems require organic matter replacement, therefore, the use of organic fertilizers has increased disregarding the fact that certain number of such fertilizers might be contaminated with the fungus, and with their handling resulting in human cases and even outbreaks of histoplasmosis. Additionally, in Colombia, chicken manure is the most common raw material used in the production of organic fertilizers. In this work, we reported the isolation of this fungus from chicken manure, and genetically compared with 42 clinical isolates. The genetically compared environmental isolates grouped together with the clinical ones. Our result suggests that chicken manure may be one of H. capsulatum infection sources. Also, the phylogenetic analyses done with other H. capsulatum isolates indicate that the Colombian isolates are widely distributed in the relational tree thus reveling towards the great genetic diversity among the H. capsulatum Colombian isolates.

Mutational analysis of the LDLR gene in a cohort of Colombian families with familial hypercholesterolemia.

BACKGROUND AND AIMS: Familial hypercholesterolemia (FH) is characterized by elevated serum cholesterol levels due to high low-density lipoprotein (LDL) cholesterol levels. FH is an autosomal dominant genetic disorder and one of the most common dominant hereditary diseases in the world. However, the frequency of mutations in Colombia is unknown. The purpose of this preliminary study was to identify mutations in the LDL receptor (LDLR) gene in a Colombian population with FH. METHODS: The study included 24 families with clinical diagnosis of sure/probable FH. The 18 exons of the LDLR were sequenced by Sanger method. RESULTS: Among 18 variants identified, 3 were known pathogenic mutations and were identified in nine individuals in five unrelated families. Five affected individuals were heterozygous for one mutation each. They were the p.W4X in two, the p.D139G in two and the p.G396D in one. Two affected individuals were homozygous for p.G396D. The variant c.1187-1G > T, which has uncertain significance in FH pathogenesis, was present in all the individuals with the p.D139G mutation. CONCLUSIONS: In total, 18 variants were identified, of which 14 correspond to known nonpathogenic variants. Three pathogenic variants were identified in the LDLR. No pathological mutations were identified in the LDLR in 79% of the study population.

Detection of Histoplasma capsulatum in Organic Fertilizers by Hc100 Nested Polymerase Chain Reaction and Its Correlation with the Physicochemical and Microbiological Characteristics of the Samples.

Histoplasma capsulatum is the causative agent of histoplasmosis and this fungus inhabits soils rich in phosphorus and nitrogen that are enriched with bird and bat manure. The replacement of organic matter in agroecosystems is necessary in the tropics, and the use of organic fertilizers has increased. Cases and outbreaks due to the presence of the fungus in these components have been reported. The Instituto Colombiano Agropecuario resolution 150 of 2003 contains the parameters set by the Colombian Technical Standard (NTC 5167) on the physicochemical and microbiological features of fertilizers, but it does not regulate the search for H. capsulatum. The aim of this study was to demonstrate H. capsulatum presence in organic fertilizers by nested polymerase chain reaction (PCR). A total of 239 samples were collected: 201 (84.1%) corresponded to organic fertilizers, 30 (12.5%) to bird excrement, and 8 (3.4%) to cave soils. The Hc100 nested PCR had a detection limit of 0.1 pg/µL and a specificity of 100%. A total of 25 (10.5%) samples were positive and validated by sequencing. Seven of the positive samples represented locations where H. capsulatum was previously detected, suggesting the persistence of the fungus. No significant correlations were detected between the physicochemical and microbiological parameters with the presence of H. capsulatum by nested PCR, indicating the fungus existence in organic fertilizers that complied with the NTC 5167. The Hc100 nested PCR targeting H. capsulatum standardized in this work will improve the evaluation of organic fertilizers and ensure the prevention of outbreaks and cases due to manufacturing, marketing, and use of fertilizers contaminated with H. capsulatum.

Epidemiology and geo-referencing of the dengue fever in a hospital of second level in Colombia, 2010-2014.

BACKGROUND: Dengue fever is a priority problem for public health in Colombia. The Department of Tolima and the municipality of El Espinal bring a considerable proportion of the cases to national statistics, becoming a representative area for studying the behavior of this pathology. OBJECTIVE: Determine the epidemiological behavior and the geographic distribution of dengue fever cases treated in the San Rafael Hospital in the municipality of the El Espinal, from 2010 to 2014. MATERIALS AND METHODS: The following is a retrospective cross-sectional case report study of the population with dengue fever treated in San Rafael Hospital. We analyzed data from the mandatory clinical reports of the SIVIGILA (National public health surveillance system, in Spanish) using Excel, EpiInfo and EpiMap were analyzed. The epidemiological analysis encompassed morbidity, mortality, fatality, endemicity and cartograms of georeferencing, among others. RESULTS: The totality of cases was 3264, with an incidence of 5.84 per 1000 inhabitants, mortality of 0.12 cases per 10,000 inhabitants, and lethality of 0.11 per 100 diagnosed cases. 71.6% of the patients were between 1 and 24 years, 53.8% were male and 46.2% female. The endemic behaviour was bimodal, Max in epidemiological periods 3-4 and 12-13. The distribution geographic of cases spanned the whole of the municipality and is related apparently with the population density in an area defined. CONCLUSIONS: The results found are above local, national and international parameters on incidence, mortality, and case-fatality. Through the analysis of endemic behavior and geographic distribution, times and specific areas are provided to optimize public health measures.

Endoscopic laser dichorionization of the placenta in the treatment of severe twin-twin transfusion syndrome.

OBJECTIVES: To describe the results of a technique of laser ablation of placental vessels in the treatment of severe twin-twin transfusion syndrome (TTTS), which is characterized by separation of the fetoplacental vascular territories and dichorionization of the placenta. PATIENTS AND METHODS: Descriptive analysis of TTTS cases treated with the endoscopic laser dichorionization of the placenta (ELDP) procedure. The variables evaluated were the occurrence of reversal of the donor-recipient phenotype, persistence of TTTS or twin anemia-polycythemia sequence (TAPS); gestational age at delivery; discharge from the hospital of at least 1 or 2 live neonates, and incidence of neurological alterations among survivors. RESULTS: 67 patients were treated with the ELDP procedure. There was no persistence of TTTS, reversal of the donor-recipient phenotype or TAPS. The median gestational age at delivery was 33.0 (23.6-37.7) weeks. The rate of discharge from the hospital of at least 1 or 2 live neonates was 88.2% (67/76) and 71.1% (54/76), respectively. Among survivors, 17 (17/121 = 14.0%) children presented with neurodevelopmental alterations during clinical follow-up. CONCLUSIONS: The major contribution of this study was the demonstration that the ELDP technique appears to be associated with a low risk of persistence or recurrence of TTTS and TAPS.

Ablación láser percutánea de un teratoma cardiaco fetal y revisión de la literatura / Percutaneous laser ablation of a foetal cardiac teratoma and a review of the literature

La ablación láser de la lesión puede enlentecer el crecimiento tumoral y la re- acumulación de líquido pericárdico. Los teratomas cardiacos son tumores benignos aunque la mitad de los fetos afectos por ellos mueren intraútero debido a taponamiento cardiaco e hidropesía fetal. En algunos casos se ha documentado una mejoría del pronóstico fetal tras el drenaje intrauterino de la efusión pericárdica. Exponemos un caso de teratoma cardiaco diagnosticado en la semana 30 de embarazo con signos de taponamiento cardiaco que fue tratado intraútero de forma percutánea mediante ablación láser de la lesión bajo control ecográfico y pericardiocentesis. El feto nació mediante cesárea electiva en la semana 35 realizándose cirugía resectiva tumoral postnatalmente(AU)

Cardiac teratomas are benign tumors, but in about half of affected fetuses there is intrauterine death due to cardiac tamponade and hydrops. In some cases there have been apparent improvements in prognosis after intrauterine drainage of the pericardial effusion. We report a case of cardiac teratoma presenting at 30 weeks gestation with cardiac tamponade, treated by ultrasound-guided laser ablation of the teratoma and pericardiocentesis. The infant was born at 35 weeks gestation and survived after post-natal tumor resection. Laser ablation of the tumor might slow down the growth and the re-accumulation of pericardial effusion(AU)

Early clinical and radiological outcomes after double osteotomy in patients with late presentation Legg-Calvé-Perthes disease.

PURPOSE: Legg-Calvé-Perthes disease is an idiopathic avascular necrosis of the femoral head. Although many surgical approaches to treat the late presentation of this pathology have been proposed, there are few reports about the early results of the double osteotomy procedure (femoral varus osteotomy combined with Salter innominate osteotomy). The purpose of this study was to describe the early results obtained with the double osteotomy in patients with late presentation of Legg-Calvé-Perthes disease. METHODS: Cross-sectional evaluation of ten patients intervened with double osteotomy. There were seven males and three females with a mean age of 9.2 +/- 1.7 years [standard deviation (SD)]. The average post-surgical time of evaluation was of 46.5 +/- 26.2 months. RESULTS: Of the ten evaluated patients, four had a Catterall III and six had a Catterall IV disease. According to Herring classification, three patients were Herring B and seven were Herring C. The epiphyseal extrusion average before and after the surgical procedure was 19.3 +/- 12.4 and 12.1 +/- 14.9%, respectively. In accordance with the Ratliff classification and Lloyd Roberts radiological results, the following were the postoperative clinical results: four good, five fair and one poor. Based on the Stulberg classification, there was one patient in class I, five in class II, three in class III and one in class IV. CONCLUSION: The surgical treatment for late Perthes disease with the best expected outcome is still a challenge. According to the resultsreported here, the double osteotomy could be considered as an alternative to treat this entity.