Analysis of COL7A1 pathogenic variants in a large cohort of dystrophic epidermolysis bullosa patients from Argentina reveals a new genotype-phenotype correlation.

Dystrophic epidermolysis bullosa (DEB) is a clinically heterogeneous heritable skin disorder, characterized by blistering of the skin and mucous membranes following minor trauma. Dominant (DDEB) and recessive (RDEB) forms are caused by pathogenic variants in COL7A1 gene. Argentina's population has a heterogeneous genetic background, and little is known about the molecular basis of DEB in our country or in native South American populations. In this study, we present the prevalence and geographical distribution of pathogenic variants found in 181 patients from 136 unrelated families (31 DDEB and 105 RDEB). We detected 95 different variants, 59 of them were previously reported in the literature and 36 were novel, nine of which were detected in more than one family. The most prevalent pathogenic variants were identified in exon 73 in DDEB patients and in exon 3 in RDEB patients. We also report a new phenotype-genotype correlation found in 10 unrelated families presenting mild blistering and severe mucosal involvement. Molecular studies in populations with an unexplored genetic background like ours revealed a diversity of pathogenic variants, and we hope that these findings will contribute to the definition of targets for new gene therapies.

The Lambda Variant in Argentina: Analyzing the Evolution and Spread of SARS-CoV-2 Lineage C.37.

The second wave of COVID-19 occurred in South America in early 2021 and was mainly driven by Gamma and Lambda variants. In this study, we aimed to describe the emergence and local genomic diversity of the SARS-CoV-2 Lambda variant in Argentina, from its initial entry into the country until its detection ceased. Molecular surveillance was conducted on 9356 samples from Argentina between October 2020 and April 2022, and sequencing, phylogenetic, and phylogeographic analyses were performed. Our findings revealed that the Lambda variant was first detected in Argentina in January 2021 and steadily increased in frequency until it peaked in April 2021, with continued detection throughout the year. Phylodynamic analyses showed that at least 18 introductions of the Lambda variant into the country occurred, with nine of them having evidence of onward local transmission. The spatial--temporal reconstruction showed that Argentine clades were associated with Lambda sequences from Latin America and suggested an initial diversification in the Metropolitan Area of Buenos Aires before spreading to other regions in Argentina. Genetic analyses of genome sequences allowed us to describe the mutational patterns of the Argentine Lambda sequences and detect the emergence of rare mutations in an immunocompromised patient. Our study highlights the importance of genomic surveillance in identifying the introduction and geographical distribution of the SARS-CoV-2 Lambda variant, as well as in monitoring the emergence of mutations that could be involved in the evolutionary leaps that characterize variants of concern.

Genetic analysis of dengue virus type 3 isolated in Buenos Aires, Argentina.

Dengue virus as a member of mosquito-borne flaviviruses is responsible for an increasing number of human infections worldwide, mainly in tropical and subtropical urban areas. The agent, a single-stranded positive sense RNA virus, is comprised of four serotypes and genetic variation within each serotype can be further divided into different genotypes. Dengue outbreaks were reported in bordering countries during the last years; the latest reported in Paraguay in 2006-2007. In Buenos Aires, 32 dengue cases were confirmed in travelers coming from this country by anti-dengue IgM antibodies detection, RT-PCR and/or isolation in C6/36 cell line. Structural proteins C, prM/M, E and non-structural proteins 1 and 2 from eight viruses were genetically characterized. Phylogenetic inference was performed for the E-protein and all viruses clustered with dengue virus 3 Genotype III. This is the first report of genetic characterization of dengue virus 3 in Argentina.

Molecular and epidemiologic analysis of enterovirus B neurological infection in Argentine children.

BACKGROUND: Human enteroviruses are one of the major causes of central nervous system (CNS) infections in pediatrics. STUDY DESIGN: We have studied 1242 children under 15 years old with suspicion of CNS infection from January 1998 to December 2003. CSF was obtained and molecular typing of human enterovirus B serotypes was performed by RT-PCR and sequencing of the N-terminal part of VP1 gene. RESULTS: According to the clinical syndromes, patients were grouped as aseptic meningitis (n=654, 52.6%), encephalitis (n=239, 19.2%), febrile seizures (n=153, 12.3%), febrile infant (n=84, 6.7%), neonatal disease (n=70, 5.6%),), acute flaccid paralysis (n=31, 2.4%) and acute disseminated encephalomyelitis (n=11, 0.9%). HEV was detected in 335/1242 CSF samples (26.97%) and was associated to aseptic meningitis (n=243, 72.5%); febrile infant (n=31, 9.2%); neonatal infection (n=26, 7.7%); encephalitis (n=25, 7.5%), febrile seizures (n=9, 2.68%); acute flaccid paralysis (n=1, 0.3%). Seasonal incidence of HEV-B species was analyzed showing that in Buenos Aires infections occur mainly during late spring and summer. Molecular serotyping was completed in 60/335 samples. Echovirus 30, Echovirus 9, Coxsackie B3 to B5 and Echovirus 33 were the most frequently identified. CONCLUSIONS: We showed that HEV are responsible for a considerable proportion of hospitalizations in children with central nervous system compromise reaching 27% of overall etiology.

Phylogenetic and molecular analyses of human parainfluenza type 3 virus in Buenos Aires, Argentina, between 2009 and 2013: The emergence of new genetic lineages.

Despite that human parainfluenza type 3 viruses (HPIV3) are one of the leading causes of acute lower respiratory tract infections in children under five, there is no licensed vaccine and there is limited current information on the molecular characteristics of regional and global circulating strains. The aim of this study was to describe the molecular characterization of HPIV3 circulating in Buenos Aires. We performed a genetic and phylogenetic analysis of the HN glycoprotein gene. Between 2009 and 2013, 124 HPIV3-positive samples taken from hospitalized pediatric patients were analyzed. Four new genetic lineages were described. Among them, C1c and C3d lineages showed local circulation patterns, whereas C3e and C3f comprised sequences from very distant countries. Despite the diversity of the described genotypes, C3a and C3d predominated over the others, the latter was present during the first years of the study and it was progressively replaced by C3a. Molecular analyses showed 28 non-synonymous substitutions; of these, 13 were located in potentially predicted B-cell epitopes. Taken together, the emergence of genetic lineages and the information of the molecular characteristics of HN protein may contribute to the general knowledge of HPIV3 molecular epidemiology for future vaccine development and antiviral therapies.

Respiratory syncytial virus: clinical and epidemiological pattern in pediatric patients admitted to a children's hospital between 2000 and 2013.

INTRODUCTION: Respiratory syncytial virus (RSV) is the major causative organism associated with acute lower respiratory tract infections in children.The objective of this study was to describe the clinical and epidemiological pattern of RSV and identify risk factors for RSV infection. POPULATION AND METHODS: Prospective, cohort study on patients hospitalized due to acute lower respiratory tract infection at Hospital de Niños Ricardo Gutiérrez between March and November throughout the 2000-2013 period. The virological diagnosis of RSV, adenovirus, influenza and parainfluenza was performed by indirect immunofluorescence using nasopharyngeal aspirates. RESULTS: A total of 12,555 children were included, 38.2% (4798) had virus rescued from samples. RSV accounted for 81.8% of cases (3924/4798) with no significant annual variations (71.2- 88.1) and with an epidemic seasonal pattern(May through July); RSV was followed by influenza (7.6%), parainfluenza (5.9%), and adenovirus (4.7%).The median age of patients with RSV rescue (3924) was 7 months old (0- 214 months old), while 74.2% were younger than 1 year old, 43.1% were younger than 6 months old, 56.5% were males and the most common clinical presentation was bronchiolitis (60.7%). Comorbidities were observed in 41.6% of cases. The most common comorbidities were chronic respiratory disease (74%), congenital heart disease (14%), and chronic neurological disease (10.2%).Complications occurred in 25%of cases. The case fatality rate was 1.9% (74/3888). Independent predictors of RSV infection were age <3 months old (OR: 2.8 [2.14-3.67], p < 0.01),clinical presentation of bronchiolitis (OR: 1.54 [1.32-1.79], p < 0.01), and hypoxemia at the time of admission (OR: 1.84 [1.42-2.37], p < 0.01). CONCLUSIONS: RSV infection displayed a seasonal pattern and was associated with infants younger than 3 months old with bronchiolitis and hypoxemia at the time of admission.

Phylogenetic and phylodynamic analyses of human metapneumovirus in Buenos Aires (Argentina) for a three-year period (2009-2011).

Human metapneumovirus, which belongs to the Paramyxoviridae family and has been classified as a member of the Pneumovirus genus, is genetically and clinically similar to other family members such as human respiratory syncytial virus. A total of 1146 nasopharyngeal aspirates from pediatric patients with moderate and severe acute lower respiratory tract infections, hospitalized at the Ricardo Gutierrez Childrens Hospital (Buenos Aires, Argentina), were tested by real time RT-PCR for human metapneumovirus. Results showed that 168 (14.65%) were positive. Thirty-six of these 168 samples were randomly selected to characterize positive cases molecularly. The phylogenetic analysis of the sequences of the G and F genes showed that genotypes A2 and B2 cocirculated during 2009 and 2010 and that only genotype A2 circulated in 2011 in Argentina. Genotype A2 prevailed during the study period, a fact supported by a higher effective population size (Neτ) and higher diversity as compared to that of genotype B2 (10.9% (SE 1.3%) vs. 1.7% (SE 0.4%), respectively). The phylogeographic analysis of the G protein gene sequences showed that this virus has no geographical restrictions and can travel globally harbored in hosts. The selection pressure analysis of the F protein showed that although this protein has regions with polymorphisms, it has vast structural and functional constraints. In addition, the predicted B-linear epitopes and the sites recognized by previously described monoclonal antibodies were conserved in all Argentine sequences. This points out this protein as a potential candidate to be the target of future humanized antibodies or vaccines.

Virus respiratorio sincicial: Patrón clínico epidemiológico en niños internados en un hospital pediátrico durante los años 2000-2013 / Respiratory syncytial virus: Clinical and epidemiological pattern in pediatric patients admitted to a children's hospital between 2000 and 2013

Introducción. El virus respiratorio sincicial (VRS) es el principal agente asociado a infección respiratoria aguda baja en niños. El objetivo de este estudio fue describir el patrón clínico-epidemiológico e identificar los factores de riesgo de infección por VRS. Población y métodos. Estudio prospectivo de cohorte de pacientes internados por infección respiratoria aguda baja en el Hospital de Niños Ricardo Gutiérrez, marzo-noviembre, 20002013. El diagnóstico viral para VRS, adenovirus, influenza y parainfluenza se realizó por inmunofluorescencia indirecta de aspirados nasofaríngeos. Resultados. Se incluyeron 12 555 niños; 38,2% (4798) presentaron rescate viral; el VRS representó el 81,8% (3924/4798) sin variaciones anuales significativas (71,2-88,1), con patrón epidémico estacional (mayo-julio); fue seguido por influenza (7,6%), parainfluenza (5,9%) y adenovirus (4,7%). Los casos con rescate de VRS (3924) tuvieron una mediana de edad de 7 meses (0-214 meses); 74,2% eran menores de 1 año; 43,1%, menores de 6 meses; 56,5%, varones; y la manifestación clínica más frecuente fue bronquiolitis (60,7%). El 41,6% tenía comorbilidades; las más frecuentes, enfermedad respiratoria crónica (74%), cardiopatías congénitas (14%) y enfermedad neurológica crónica (10,2%). El 25% presentó complicaciones. La letalidad fue 1,9% (74/3888). Los predictores independientes de infección por VRS fueron la edad < 3 meses OR 2,8 (2,14-3,67), p < 0,01 , la bronquiolitis como presentación clínica OR 1,54 (1,32-1,79), p < 0,01 y la presencia de hipoxemia al momento del ingreso OR 1,84 (1,42-2,37), p < 0,01 . Conclusiones. La infección por VRS presentó un patrón epidémico estacional y se asoció más a niños pequeños menores de tres meses con bronquiolitis e hipoxemia al momento del ingreso.

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