Differential virome composition and richness between children's diarrheagenic stools kept at ultra-low temperatures for long-term.

INTRODUCTION: Diarrhoeal illness is the second cause of morbidity/mortality among children from less-developed regions worldwide. Nonetheless, there is scarce information regarding their gut microbiome. AIM: Microbiome characterization, with an emphasis on the virome, of children's stools with diarrhoea, by a commercial microbiome array. METHODOLOGY: Nucleic acids extraction, optimised for viral identification, of stool samples from 20 Mexican children with diarrhoea (10 children < 2 and 10 ≥ 2-years-old), collected 16 years ago and kept at -70 °C, were analysed for the presence of viruses, bacteria, archaea, protozoa, and fungi species sequences. RESULTS: Only viral and bacterial species sequences were identified among children's stools. Most stool samples harboured species belonging to the bacteriophages (95%), anellovirus (60%), diarrhoeagenic viruses (40%), and non-human pathogens viruses (45% avian virus and 40% plant viruses) groups. Among the children's stools, virome inter-individual species composition was observed, even in presence of illness. The < 2-years-old children group has significantly higher viral richness (p = 0.01), conferred mainly by bacteriophages and diarrheagenic-viruses (p = 0.01) species, in comparison with the ≥ 2-years-old group. CONCLUSIONS: The virome of stools of children with diarrhoea revealed inter-individual viral species composition. Similarly, to the few virome studies in healthy young children, the bacteriophages group was the most abundant. A significantly higher viral richness, conferred by bacteriophages and diarrheagenic-viral species, was observed among < 2-years-old children in comparison with older children. Stools preserved at -70 °C for long term can successfully be used for microbiome studies.

Neuronal ceroid lipofuscinosis in the South American-Caribbean region: An epidemiological overview.

Neuronal ceroid lipofuscinoses (NCLs) comprise 13 hereditary neurodegenerative pathologies of very low frequency that affect individuals of all ages around the world. All NCLs share a set of symptoms that are similar to other diseases. The exhaustive collection of data from diverse sources (clinical, genetic, neurology, ophthalmology, etc.) would allow being able in the future to define this group with greater precision for a more efficient diagnostic and therapeutic approach. Despite the large amount of information worldwide, a detailed study of the characteristics of the NCLs in South America and the Caribbean region (SA&C) has not yet been done. Here, we aim to present and analyse the multidisciplinary evidence from all the SA&C with qualitative weighting and biostatistical evaluation of the casuistry. Seventy-one publications from seven countries were reviewed, and data from 261 individuals (including 44 individuals from the Cordoba cohort) were collected. Each NCL disease, as well as phenotypical and genetic data were described and discussed in the whole group. The CLN2, CLN6, and CLN3 disorders are the most frequent in the region. Eighty-seven percent of the individuals were 10 years old or less at the onset of symptoms. Seizures were the most common symptom, both at onset (51%) and throughout the disease course, followed by language (16%), motor (15%), and visual impairments (11%). Although symptoms were similar in all NCLs, some chronological differences could be observed. Sixty DNA variants were described, ranging from single nucleotide variants to large chromosomal deletions. The diagnostic odyssey was probably substantially decreased after medical education activities promoted by the pharmaceutical industry and parent organizations in some SA&C countries. There is a statistical deviation in the data probably due to the approval of the enzyme replacement therapy for CLN2 disease, which has led to a greater interest among the medical community for the early description of this pathology. As a general conclusion, it became clear in this work that the combined bibliographical/retrospective evaluation approach allowed a general overview of the multidisciplinary components and the epidemiological tendencies of NCLs in the SA&C region.

A Novel Adult Murine Model of Typical Enteroaggregative Escherichia coli Infection Reveals Microbiota Dysbiosis, Mucus Secretion, and AAF/II-Mediated Expression and Localization of ß-Catenin and Expression of MUC1 in Ileum.

Typical enteroaggregative Escherichia coli (tEAEC) is a diarrheagenic E. coli pathotype associated with pediatric and traveler's diarrhea. Even without diarrhea, EAEC infections in children also lead to increased gut inflammation and growth shortfalls. EAEC strain's defining phenotype is the aggregative adherence pattern on epithelial cells attributable to the aggregative adherence fimbriae (AAF). EAEC only causes diarrhea in humans; therefore, not much is known of the exact intestinal region of infection and damage or its interactions with intestinal enterocytes in vivo and in situ. This study aimed to develop a new tEAEC mouse model of infection, characterize the microbiota of infected mice, and evaluate in situ the expression of host adherence and surface molecules triggering EAEC infection and the role of the EAEC AAF-II in adherence. Six-week-old C57BL/6 mice, without previous antibiotic treatment, were orally challenged with EAEC 042 strain or EAEC 042 AAF-II mutant (ΔAAF/II) strain, or DAEC-MXR strain (diffusely adherent E. coli clinical isolate), and with saline solution (control group). Paraffin sections of the colon and ileum were stained with H&E and periodic acid-Schiff. ZO-1, ß-catenin, MUC1, and bacteria were analyzed by immunofluorescence. EAEC-infected mice, in comparison with DAEC-MXR-infected and control mice, significantly lost weight during the first 3 days. After 7 days post-infection, mucus production was increased in the colon and ileum, ZO-1 localization remained unaltered, and morphological alterations were more pronounced in the ileum since increased expression and apical localization of ß-catenin in ileal enterocytes were observed. EAEC-infected mice developed dysbiosis 21 days post-infection. At 4 days post-infection, EAEC strain 042 formed a biofilm on ileal villi and increased the expression and apical localization of ß-catenin in ileal enterocytes; these effects were not seen in animals infected with the 042 ΔAAF/II strain. At 3 days post-infection, MUC1 expression on ileal enterocytes was mainly detectable among infected mice and colocalized with 042 strains on the enterocyte surface. We developed a novel mouse model of EAEC infection, which mimics human infection, not an illness, revealing that EAEC 042 exerts its pathogenic effects in the mouse ileum and causes dysbiosis. This model is a unique tool to unveil early molecular mechanisms of EAEC infection in vivo and in situ.

Immune Alterations in a Patient With Hyperornithinemia-Hyperammonemia-Homocitrullinuria Syndrome: A Case Report.

The hyperornithinemia-hyperammonemia-homocitrullinuria (HHH) syndrome is a rare autosomal recessive inborn error of the urea cycle caused by mutations in the SLC25A15 gene. Besides the well-known metabolic complications, patients often present intercurrent infections associated with acute hyperammonemia and metabolic decompensation. However, it is currently unknown whether intercurrent infections are associated with immunological alterations besides the known metabolic imbalances. Herein, we describe the case of a 3-years-old girl affected by the HHH syndrome caused by two novel SLC25A15 gene mutations associated with immune phenotypic and functional alterations. She was admitted to the hospital with an episode of recurrent otitis, somnolence, confusion, and lethargy. Laboratory tests revealed severe hyperammonemia, elevated serum levels of liver transaminases, hemostasis alterations, hyperglutaminemia and strikingly increased orotic aciduria. Noteworthy, serum protein electrophoresis showed a reduction in the gamma globulin fraction. Direct sequencing of the SLC25A15 gene revealed two heterozygous non-conservative substitutions in the exon 5: c.649G>A (p.Gly217Arg) and c.706A>G (p.Arg236Gly). In silico analysis indicated that both mutations significantly impair protein structure and function and are consistent with the patient clinical status confirming the diagnosis of HHH syndrome. In addition, the immune analysis revealed reduced levels of serum IgG and striking phenotypic and functional alterations in the T and B cell immune compartments. Our study has identified two non-previously described mutations in the SLC25A15 gene underlying the HHH syndrome. Moreover, we are reporting for the first time functional and phenotypic immunologic alterations in this rare inborn error of metabolism that would render the patient immunocompromised and might be related to the high frequency of intercurrent infections observed in patients bearing urea cycle disorders. Our results point out the importance of a comprehensive analysis to gain further insights into the underlying pathophysiology of the disease that would allow better patient care and quality of life.

"Atypical" Phenotypes of Neuronal Ceroid Lipofuscinosis: The Argentine Experience in the Genomic Era

ABSTRACT Neuronal Ceroid Lipofuscinosis (NCL) refers to a group of inherited lysosomal storage disorders characterized by the intracellular accumulation of ceroid-lipofuscin compounds and neurodegeneration. Fourteen genes are currently recognized with disease-causing DNA variants: PPT1/CLN1, TPP1/CLN2, CLN3, DNAJC5/CLN4, CLN5, CLN6, MFSD8/CLN7, CLN8, CTSD/CN10, GRN/CLN11, ATP13A2/CLN12, CTSF/CLN13, KCTD7/CLN14, TBCK/CLN15. In the frame of the Cordoba cohort, we studied N=51 cases. The aim of this paper is the observational and retrospective analysis of the "atypical" phenotypes. PCR-Sanger sequencing and/or massive exome sequencing were used as a screening methodology. One CLN1 subject showed an atypical prolonged (P) phenotype with null PPT1 activity and a heterozygous compound genotype: E5 c.451C>T, p.Arg151*/g.6302T>G (I3 c.363-3T>G). Other 11 CLN2 individuals (except one girl) showed TPP1 activity decreased to around 10% of the minimum value of the reference interval in leukocytes and saliva. The DNA variants E7 c.827A>T, p.Asp276Val and I7 c.887-10A>G were the most prevalent. One CLN8 individual showed an atypical congenital phenotype with a heterozygous combination of DNA variants: E2 c.1A>G, p.?/E3 c.792C>G, p.Asn264Lys. Massive sequencing was installed as a screening methodology for the precision diagnosis of atypical CLN1, CLN2, and CLN8 phenotypes. A genetic/phenotypic local registry is under construction.

Lesch-Nyhan Disease and Its Variants: Phenotypic and Mutation Spectrum of Hypoxanthine-Guanine Phosphoribosyltransferase Deficiency in Argentine Patients

Abstract Hypoxanthine-guanine phosphoribosyltransferase (HPRT) deficiency is a disorder of purine metabolism responsible for Lesch-Nyhan Disease (LND) and its variants, HPRT-related hyperuricemia with neurologic dysfunction (HND) and HPRT-related hyperuricemia (HRH). The objective of this study was to characterize a cohort of Argentine patients with HPRT deficiency diagnosed in a single center. Results: Twenty nine patients were studied, including 12 LND, 15 HND and 2 HRH. The average onset age was 0.64 years for LND with motor delay as the main manifestation, 8.84 years for HND and 2.5 years for HRH; nephrological manifestations predominated as presenting features in these variants. The average diagnosis age was 3.58 years for LND, 17.21 years for HND and 2.5 years for HRH. Clinical heterogeneity was more evident in HND, even in members of the same family. All patients presented hyperuricemia and no detectable HPRT activity in erythrocyte lysate. The molecular study allowed to identify 9 different mutations in HPRT1 gene from 24 patients (11 independent pedigrees) and to establish genotype-phenotype correlation. In conclusion, this study describes the genotypic/phenotypic spectrum of HPRT deficiency in Argentine patients and highlights the need to increase awareness about the suspicion of these diseases, especially the LND variants with high clinical heterogeneity.

Estado poblacional del cucarachero de pantano, Cistothorus apolinari (Passeriformes: Troglodytidae) en siete humedales de la Sabana de Bogotá, Colombia

Introduction: The Apolinar's Wren Cisthotorus apolinari is an endemic species of the Eastern Andes of Colombia currently classified as Critically Endangered (CR) at the national level and Endangered (EN) worldwide, mainly due to the degradation of wetlands, their primary habitat, and the parasitism of the nests. Objetive: Evaluate the state of the populations of C. apolinari in seven wetlands of the Sabana de Bogotá, searching to define what the areas evaluated mean that it hosts the largest population of the species and what other factors determine these population sizes. Methods Between July and December 2014 in seven wetlands we monitoring carried out using counting points and auditory censuses, the abundance of the Apolinar's Wren was recorded, the vegetation cover where the individuals were recorded and the abundances of Shiny Cowbird Molothrus bonariensis. Results: There were 63.6 h of observation and 88 counting points, obtain a nine records of C. apolinari , one individual in Tibanica, three in La Florida and five in Gualí, mainly associated with the reed Schoenoplectus californicus. None of the biotic and abiotic factors evaluated in the wetlands, were found to determine the presence of the Apolinar's Wren, but trends were present for some variables as the presence of S. californicus and Thypa spp. Conclusions: The population of C. apolinari has had a significant reduction of up to 94 %, added to a possible local extinction in the wetland of La Conejera. These results seek to provide relevant information to contribute to the formulation of effective conservation measures for the protection of the species and its habitat throughout its distribution in the country.

Urea cycle disorders in Argentine patients: clinical presentation, biochemical and genetic findings.

BACKGROUND: The incidence, prevalence, and molecular epidemiology of urea cycle disorders (UCDs) in Argentina remain underexplored. The present study is the first to thoroughly assess the clinical and molecular profiles of UCD patients examined at a single reference center in Argentina. RESULTS: Forty-nine UCD cases were collected. About half (26/49, 53%) manifested neonatally with classical presentation and had a high mortality (25/26, 96%). Ornithine transcarbamylase deficiency (OTCD) was the most common UCD (26 patients). Argininosuccinate synthetase deficiency (ASSD) was detected in 19 cases, while argininosuccinate lyase deficiency (ASLD) was diagnosed in 4 cases. Molecular genetic analysis revealed 8 private OTC mutations and two large deletion/duplication events in the OTC gene. Most mutations in the ASS1 and ASL genes were recurrent missense changes, and four alterations were novel. The clinical outcome of our UCD cohort was poor, with an overall mortality of 57% (28/49 cases), and a 28% (6/21) disability rate among the survivors. CONCLUSIONS: Most patients in our case series showed severe neonatal onset, with high morbidity/mortality. We detected in total 19 mutations, most of them recurrent and of high frequency worldwide. Noteworthy, we highlight the presence of a geographic cluster with high prevalence of a point mutation in the ASS1 gene. This study suggests that these disorders may be more frequent than commonly assumed, and stresses the need for increased awareness amongst health professionals and greater availability of diagnostic tools for accurate identification, early diagnosis, and timely treatment.

Draft Genome Sequence of Escherichia albertii Strain Mex-12/320a, Isolated from an Infant with Diarrhea and Harboring Virulence Genes Associated with Diarrheagenic Strains of Enteropathogenic Escherichia coli.

Escherichia albertii is an emerging human enteropathogen. We report the draft genome sequence of E. albertii strain Mex-12/320a, isolated from an infant with diarrhea. The presence of the pathogenic island O122/IE6 and the nleA gene, previously found in diarrheagenic enteropathogenic Escherichia coli strains, suggests that E. albertii may cause acute diarrhea.

Estenosis glótica por sinequia de cuerdas vocales secundario a cuerpo extraño: A propósito de un caso / Stenosis glottal synechia of vocal cords for secondary to foreign body: About a case

El cuerpo extraño en vía aérea es una patología frecuente. Se presenta como un evento súbito en un niño previamente sano con manifestación de dificultad respiratoria severa. Puede ser un incidente anodino o provocar complicaciones graves. Dentro de las cuales encontramos la estenosis glótica y subglótica. Esta última puede ser congénita o adquirida, la adquirida se presenta con frecuencia posterior a intubación endotraqueal, trauma directo por cuerpo extraño o trauma externo. El tratamiento depende de su localización, naturaleza y lesiones asociadas. Describimos en este caso un lactante de 8 meses que ingiere de forma accidental cuerpo extraño, el cual se extrae de forma tardía (más de 48 horas) presentando posteriormente dificultad respiratoria, estridor y disfonía, se realiza estudio endoscópico laríngeo evidenciándose estenosis subglótica inicialmente sin afectación de cuerdas vocales, en vista de evolución tórpida en control imagenologico se evidencia afección severa de cuerdas vocales, poco frecuente en la edad pediátrica. Ameritando microcirugía laríngea.

Foreign bodies in the airway is a common condition. It comes as a sudden event in a previously healthy child with manifestation of severe respiratory distress. It may be a nondescript incident and cause serious complications such as glottic and subglottic stenosis. The latter can be congenital or acquired, the acquired type is a frequent late presentation of endotracheal intubation, direct trauma by foreign body or external trauma. Treatment depends on the location, nature and associated injuries. We describe an 8 months infant who accidentally swallowed a foreign body, which was extracted after 48 hours. Posteriorly he presented difficult breathing, stridor and dysphonia. Laryngeal endoscopic study demonstrated a subglottic stenosis with unaffected vocal cord. In view of a torpid evolution, control endoscopic study reported severe vocal cord lesion which is a rare finding in children. Laryngeal microsurgery was required for his resolution.

The neuronal ceroid lipofuscinoses program: A translational research experience in Argentina.

BACKGROUND: The Argentinean program was initiated more than a decade ago as the first experience of systematic translational research focused on NCL in Latin America. The aim was to overcome misdiagnoses and underdiagnoses in the region. SUBJECTS: 216 NCL suspected individuals from 8 different countries and their direct family members. METHODS: Clinical assessment, enzyme testing, electron microscopy, and DNA screening. RESULTS AND DISCUSSION: 1) The study confirmed NCL disease in 122 subjects. Phenotypic studies comprised epileptic seizures and movement disorders, ophthalmology, neurophysiology, image analysis, rating scales, enzyme testing, and electron microscopy, carried out under a consensus algorithm; 2) DNA screening and validation of mutations in genes PPT1 (CLN1), TPP1 (CLN2), CLN3, CLN5, CLN6, MFSD8 (CLN7), and CLN8: characterization of variant types, novel/known mutations and polymorphisms; 3) Progress of the epidemiological picture in Latin America; and 4) NCL-like pathology studies in progress. The Translational Research Program was highly efficient in addressing the misdiagnosis/underdiagnosis in the NCL disorders. The study of "orphan diseases" in a public administrated hospital should be adopted by the health systems, as it positively impacts upon the family's quality of life, the collection of epidemiological data, and triggers research advances. This article is part of a Special Issue entitled: "Current Research on the Neuronal Ceroid Lipofuscinoses (Batten Disease)".

Trends in frequency and prevalence of oral cancer and oral squamous cell carcinoma in Mexicans. A 20 years retrospective study.

OBJECTIVE: To establish the time trends of the frequency and prevalence of oral cavity cancer in regard to age and gender in a 20-years (time period 1989 - 2008) cohort of Mexicans. DESIGN AND SETTING: 13,235 head and neck biopsies from the archive of the Oral Pathology Laboratory, Dental School, National Autonomous University of Mexico were revised. The cases with diagnoses of oral cancer were selected. Gender and age at diagnosis was obtained from medical records. The frequency and prevalence of oral cavity cancer and oral squamous cell carcinoma were assessed biannually in regard to the total number of population served by the oral pathology laboratory. The statistical significance of trends was established using the linear logistic regression (curve estimation) test (s 0.05). RESULTS: 298 cases (138 males; 160 females) of oral cancer were included; 167 (92 females; 75 males; female:male ratio: 1.1:1) corresponded to oral squamous cell carcinoma. From 1989 to 2008 the prevalence of oral cancer and oral squamous cell carcinoma increased 200% (s 0.05) and 100% (s 0.000) respectively. The increase of frequency and prevalence was observed in both genders however only in females was significant (s 0.000). We do not identify changes in the age at diagnosis. CONCLUSIONS: Oral cancer, specifically oral squamous cell carcinoma, has increase in Mexicans females in the last 20 years.

Errores frecuentes en la administración de medicamentos intravenosos en pediatría / Common errors in administration of intravenous medications in pediatrics

Introducción: A nivel mundial se reportan errores en la administración de medicamentos intravenosos, afortunadamente son excepcionales los casos graves. Objetivo: Identificar los errores más frecuentes del personal de enfermería en la administración de medicamentos intravenosos en pacientes pediátricos en una unidad de alta especialidad. Metodología: Estudio transversal, observacional, descriptivo, se estudiaron por observación con lista de verificación a enfermeras generales y especialistas que realizaron el procedimiento de administración de medicamentos intravenosos, se midió conocimiento con un cuestionario previamente validado por consenso de expertos. El análisis de datos se llevó a cabo con estadística descriptiva. Resultados: De 230 procedimientos se identificaron errores de registro (43%), donde el personal de enfermería no realizó el registro inmediatamente, error de preparación del fármaco, entendido como dosis inexacta (31%), al evaluar conocimiento (68%) de las enfermeras tuvieron conocimiento "eficiente" y (32%) conocimiento deficiente. Discusión: Los errores en la administración de medicamentos mostraron un comportamiento similar a lo referido en la literatura, sin embargo este estudio mostró que el error más frecuente fue el registro porque se realizó en diferentes momentos y mostró ligeramente un porcentaje menor a lo referido en la literatura, el error de preparación del fármaco tuvo 2% menos a lo referido por otro autor. Conclusión: A lo que se llamó errores en este trabajo, únicamente quedaron en "cuasi-fallas" tipo 2 que no llegaron a causar ningún daño al paciente. Sin embargo es necesario introducir mejoras en la organización de los servicios que se traduzcan en una práctica clínica segura.

Introduction: At worldwide level mistakes during the administration of intravenous medications are reported, fortunately, are exceptional the number of hazard cases. Objective: To identify the most frequent mistakes from nursing staff during the administration of intravenous medications in pediatrics in a specialized medical facility. Methodology: Transversal, observational, and descriptive study. Registered and specialist nurses were studied through a check list meanwhile they were doing a procedure of administration of intravenous medications. Their knowledge was measured through a questionnaire previously validated by expert consensus. Data analysis was done with descriptive statistics. Results: From 230 procedures, recording errors were identified (43%) in which nursing staff did not make the record immediately after; error when setting up the medication to be administered, meaning as inexact dose (31%). When evaluating the nurses' knowledge 68% nurses had "efficient" and 32% deficient knowledge. Discussion: Mistakes during the administration of medications showed a similar performance to that showed in medical literature; however, this study showed that the most frequent mistake was recording because it was done at different times and showed a percentage lightly minor to that referred in the medical literature. The error when setting up the medication had 2% lees to that referred in the medical literature by another author. Conclusions: In this research errors mean "pseudo-fails" type 2 which did not mean to be hazardous to patients; however, it is necessary inclusion of improvements within the organization of services that show up a safer clinical practice.