The correlation of two different real-time PCR devices for the analysis of CYP2C19 pharmacogenetic results

CYP2C19 is a highly polymorphic gene responsible for the metabolism of commonly used drugs. CYP2C19*1, the wild-type allele, is associated with normal enzyme activity, whereas CYP2C19*2 and CYP2C19*17 lead to null and increased enzyme activity, respectively. The use of different instruments to perform the same pharmacogenetic tests should not affect the reliability of the results reported to clinicians, as required by the ISO 15189 standard. Genotyping assays allowed for the identification of gene variants corresponding to the CYP2C19*2 and CYP2C19*17 haplotypes in 44 selected samples. Each sample was analyzed in duplicate using the Thermo Fisher Taqman Drug Metabolism probes CYP2C19*2: c_25986767_70 (rs4244285) and CYP2C19*17: c_469857_10 (rs12248560). The experiments were performed on two widely used types of real-time PCR analyzers: ABI PRSIM™7500 and QuantStudioTM12KFlex (both from Applied Biosystems, Thermofisher). The data were analyzed in a Thermo Fisher Cloud facility. The analysis was performed independently by two qualified professionals. Both different instruments and analysts’ interpretations were consistent in identifying the native homozygous, heterozygous, and mutant homozygous variants for CYP2C19*2 and CYP2C19*17. The results provided by both the primary and backup analyzers showed a perfect correlation. This would allow for the use of the backup analyzer in case the main one is not available

Online calculator for individual affiliation to a major population group

Because of their sensitivity and high level of discrimination, short tandem repeat (STR) maker systems are currently the method of choice in routine forensic casework and data banking, usually in multiplexes up to 15–17 loci. Constraints related to sample amount and quality, frequently encountered in forensic casework, will not allow to change this picture in the near future, notwithstanding the technological developments. In this study, we present a free online calculator named PopAffiliator (http://cracs.fc.up.pt/popaffiliator) for individual population affiliation in the three main population groups, Eurasian, East Asian and sub-Saharan African, based on genotype profiles for the common set of STRs used in forensics. This calculator performs affiliation based on a model constructed using machine learning techniques. The model was constructed using a data set of approximately fifteen thousand individuals collected for this work. The accuracy of individual population affiliation is approximately 86%, showing that the common set of STRs routinely used in forensics provide a considerable amount of information for population assignment, in addition to being excellent for individual identification

Continent-wide decoupling of Y-chromosomal genetic variation from language and geography in native South Americans

Numerous studies of human populations in Europe and Asia have revealed a concordance between their extant genetic structure and the prevailing regional pattern of geography and language. For native South Americans, however, such evidence has been lacking so far. Therefore, we examined the relationship between Y-chromosomal genotype on the one hand, and male geographic origin and linguistic affiliation on the other, in the largest study of South American natives to date in terms of sampled individuals and populations. A total of 1,011 individuals, representing 50 tribal populations from 81 settlements, were genotyped for up to 17 short tandem repeat (STR) markers and 16 single nucleotide polymorphisms (Y-SNPs), the latter resolving phylogenetic lineages Q and C. Virtually no structure became apparent for the extant Y-chromosomal genetic variation of South American males that could sensibly be related to their inter-tribal geographic and linguistic relationships. This continent-wide decoupling is consistent with a rapid peopling of the continent followed by long periods of isolation in small groups. Furthermore, for the first time, we identified a distinct geographical cluster of Y-SNP lineages C-M217 (C3*) in South America. Such haplotypes are virtually absent from North and Central America, but occur at high frequency in Asia. Together with the locally confined Y-STR autocorrelation observed in our study as a whole, the available data therefore suggest a late introduction of C3* into South America no more than 6,000 years ago, perhaps via coastal or trans-Pacific routes. Extensive simulations revealed that the observed lack of haplogroup C3* among extant North and Central American natives is only compatible with low levels of migration between the ancestor populations of C3* carriers and non-carriers. In summary, our data highlight the fact that a pronounced correlation between genetic and geographic/cultural structure can only be expected under very specific conditions, most of which are likely not to have been met by the ancestors of native South Americans

Proyecto, investigación e innovación en urbanismo, arquitectura y diseño industrial

Actas de congresoLas VII Jornadas de Investigación “Encuentro y Reflexión” y I Jornadas de Investigación de becarios y doctorandos. Proyecto, investigación e innovación en Urbanismo, Arquitectura y Diseño Industrial se centraron en cuatro ejes: el proyecto; la dimensión tecnológica y la gestión; la dimensión social y cultural y la enseñanza en Arquitectura, Urbanismo y Diseño Industrial, sustentados en las líneas prioritarias de investigación definidas epistemológicamente en el Consejo Asesor de Ciencia y Tecnología de esta Universidad Nacional de Córdoba. Con el objetivo de afianzar continuidad, formación y transferencia de métodos, metodología y recursos se incorporó becarios y doctorandos de los Institutos de investigación. La Comisión Honoraria la integraron las tres Secretarias de Investigación de la Facultad, arquitectas Marta Polo, quien fundó y María del Carmen Franchello y Nora Gutiérrez Crespo quienes continuaron la tradición de la buena práctica del debate en la cotidianeidad de la propia Facultad. Los textos que conforman las VII Jornadas son los avances y resultados de las investigaciones realizadas en el bienio 2016-2018.Fil: Novello, María Alejandra. Universidad Nacional de Córdoba. Facultad de Arquitectura, Urbanismo y Diseño; ArgentinaFil: Repiso, Luciana. Universidad Nacional de Córdoba. Facultad de Arquitectura, Urbanismo y Diseño; ArgentinaFil: Mir, Guillermo. Universidad Nacional de Córdoba. Facultad de Arquitectura, Urbanismo y Diseño; ArgentinaFil: Brizuela, Natalia. Universidad Nacional de Córdoba. Facultad de Arquitectura, Urbanismo y Diseño; ArgentinaFil: Herrera, Fernanda. Universidad Nacional de Córdoba. Facultad de Arquitectura, Urbanismo y Diseño; ArgentinaFil: Períes, Lucas. Universidad Nacional de Córdoba. Facultad de Arquitectura, Urbanismo y Diseño; ArgentinaFil: Romo, Claudia. Universidad Nacional de Córdoba. Facultad de Arquitectura, Urbanismo y Diseño; ArgentinaFil: Gordillo, Natalia. Universidad Nacional de Córdoba. Facultad de Arquitectura, Urbanismo y Diseño; ArgentinaFil: Andrade, Elena Beatriz. Universidad Nacional de Córdoba. Facultad de Arquitectura, Urbanismo y Diseño; Argentin

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Análisis molecular de variación de polimorfismos STR autosómicos y de cromosoma «Y» en grupos étnicos de Ecuador

The results found allow us to establish certain conclusions on the structure of the population and the origin of Ecuadorians today. The volume of information obtained and the knowledge acquired continue to be insufficient in order to understand this ethnical, geographical and genetic structure of New World settlers. The genetic studies completed represent the first formal research into Ecuadorian ethno-genetics. The cultural crossbreeding can be found in different issues like economic, social, political and religious organizations, linguistic systems, communication codes, etc. The ethnic mixture process initiated during colonialisation continues today. The dominant economic and political group conveys the culture which is imposed in its economic, social, political and linguistic models for the others groups. The dominant culture leads the development of the Western culture and tends towards uniformity. It is presented as the "cultura ideal" and is transmitted by the media and in daily interaction. Ecuador is clearly a multi-ethnic and multicultural country. Cultural assessment, respect and rescue do not mean assessing, respecting and rescuing isolated cultural elements, but rather social groups with rights to resources for subsistence, to political representation and their own social and organizational bodies, to education in their own language and culture, and to determine their own development in line with their needs. In this work, we analyzed samples from three different ethnic groups of Ecuador, including Mestizos, indigenous American Kichwas and Afro-American Blacks; analysis included 20 autosomic microsatellites D3S1358, HumFGA, D21S11, Penta E, Penta D, HumVWA, D8S1179, D7S820, D13S317, D5S818, D16S539, HumTH01, HumCSF1PO, HumTPOX, HumF13A01, HumF13B, HumLPL, HumHPRTB, HumFES/FPS and amelogenin; there was also capillary electrophoresis analysis of 12 microsatellites of the Y-chromosome such as DYS19, DYS385 a/b, DYS 389I, DYS389II, DYS390, DYS391, DYS392, DYS392, DYS393, DYS437, DYS438 and DYS439. With the autosomal STR markers we have confirmed and quantified that Mestizos and Afro-Ecuadorians are tri-hybrid populations. Mestizos contain ~73% of autosomal chromosomes of Native Americans origin, ~19% putative European and ~8% African origin. Afro-Ecuadorians can be estimated at ~57% Africans, ~28% Europeans and ~15% Native Americans. Seven different haplotypes were shared between Kichwas and Mestizos, one between Mestizos and Afro-Ecuadorians and one between Kichwas, Afro-Ecuadorians and Mestizos. This latter one is the most frequent haplotype in Europeans and, in particular, in Spaniards. The total number of different haplotypes is 276 in the entire sample analyzed. Mestizos are quite close to Kickwas, but are clearly closer to Spaniards than Kichwas. It is also the case for respective distance to Guineans. This is consistent with a triple genetic origin for the Mestizos: Amerinds, Europeans and Africans, as shown by the STRs in the Y-chromosome. Afro-Ecuadorians are the closest to Guineans, but are closer to Kichwas and to Spaniards than Guineans. With different proportions of mixture, the triple origin model proposed for the Mestizos is also applicable to Afro-Ecuadorians. The haplotypic heredity and the large possibility of combinations which these markers offer warn of the possible existence of allelic compositions as yet unanalyzed, and of those in which the real frequency is unknown.La Genética Forense es una de las ramas de la biomedicina que ha mostrado un vertiginoso desarrollo científico. A raíz de este crecimiento, numerosas organizaciones internacionales oficiales han adoptado sistemas de análisis de ADN que pudieran ser transferidos y equiparados en distintos países, para favorecer el intercambio de información científica, técnica y pericial, sobre todo en la resolución de caso judiciales como estudios de paternidad y de criminalística forense. Dicho trabajo no podría ser ejecutado sin contar con Bases de Datos poblacionales amplias y adecuadamente estudiadas. En este trabajo se analizaró un total de 1438 muestras de 4 grupos étnicos de Ecuador, entre ellos mestizos, amerindios kichwas, amerindios waorani y negros afroamericanos; se analizó por técnicas manuales y electroforesis capilar 20 microsatélites autonómicos D3S1358, HumFGA, D21S11, Penta E, Penta D, HumVWA, D8S1179, D7S820, D13S317, D5S818, D16S539, HumTH01, HumCSF1PO, HumTPOX, HumF13A01, HumF13B, HumLPL, HumHPRTB, HumFES/FPS y amelogenina; se analizaron también por electroforesis capilar 12 microsatélites del cromosoma Y como DYS19, DYS385 a/b, DYS 389I, DYS389II, DYS390, DYS391, DYS392, DYS392, DYS393, DYS437, DYS438 y DYS439. Todos los grupos étnicos analizados mostraron estar en equilibrio de Hardy-Weinberg para todos los STRs autosómicos analizados. La utilización en forma conjunta de 15 STRs autosómicos (en formato multiplex) ha demostrado poseer una elevada variabilidad y ofrece un Poder de Discriminación (PD) conjunto superior a 0,99999999 y de Poder de Exclusión a priori (MEC) siempre por encima de 0,99999925, la Probabilidad de Matching combinado encontrado es de 1 en 1.27 x 1017 para mestizos. Se ha confirmado y cuantificado que mestizos y afroecuatorianos son poblaciones trihíbridas, con proporciones variables de contribución de amerindios, europeos y africanos. Como se observa a partir de cromosoma "Y", la contribución de los varones europeos (claramente los españoles en el caso de los mestizos) fue mucho mayor que cuando las estimaciones de mezcla se calcularon a partir de los STRs autosómicos. Se ha confirmado y cuantificado con los marcadores STRs autosómicos que mestizos y afroecuatorianos son poblaciones trihíbridas. Los mestizos contienen ~73% de cromosomas autosómicos de origen amerindio, un ~19% europeo putativo y ~8% origen africano. Los afroecuatorianos se pueden estimar en el ~57% africanos, ~28% europeos y ~15% nativos americanos. En mestizos, amerindios kichwas y negros afroamericanos, la diversidad haplotípica del cromosoma "Y" es alta y bastante próxima a uno; se debe observar que este parámetro, en los sistemas haploides como ADN mitocondial y el cromosoma "Y", es numéricamente idéntico a los parámetros forenses de información a priori como el poder de discriminación o el poder de exclusión en los casos de paternidad. Se confirma que los mestizos tienen una corta distancia genética con los españoles y con las poblaciones caucasoides de la península ibérica, mientras que los kichwas claramente nativos del nuevo continente se aproximan a la mayoría de poblaciones urbanas de centroamérica y del área andina, en aquellos sitios donde existe una clara presencia indígena. Siete haplotipos diferentes fueron compartidos entre kichwas y mestizos, uno entre mestizos y afroecuatorianos y uno entre kichwas, afroecuatorianos y mestizos. Este último resulta ser el haplotipo más frecuente en los europeos y, particularmente, en los españoles. El número total de haplotipos diferentes es 276 en toda la muestra analizada. Los mestizos muestran una distancia corta a los kickwas, pero su distancia a los españoles es claramente más corta que entre los kichwas y los españoles. También es este el caso para su respectiva distancia a los guineanos. Esto es consistente con un triple origen genético para los mestizo: amerindios, europeos y africanos, como se demuestra por los STRs en el cromosoma "Y" . Los afroecuatorianos son los más próximos a los guineanos pero están más próximos a los kichwas y a los españoles que lo que lo están los guineanos. Con diferentes proporciones de mezcla, el modelo de triple origen propuesto para los mestizos también aplica a los afroecuatorianos. La herencia haplotípica y la gran posibilidad de combinaciones que ofrecen estos marcadores advierte de la posible existencia de composiciones alélicas todavía sin analizar, y de aquellas en las que se desconoce la frecuencia real. La colaboración de todos los laboratorios forenses permitirá seguir ampliando la base de datos para el cromosoma Y, de este modo se podrá aumentar la precisión estadística a la hora de interpretar los resultados de la investigación de la individualidad biológica en la rutina forense. Los datos obtenidos son de enorma valor antropológico y permitirán trazar mejor la historia de Ecuador en busca de los orígnes de los actuales pobladores del Nuevo Mundo

A comprehensive study of Ecuadorian adult patients with a mild and moderate presentation of COViD-19.

AimTo characterize non-hospitalized patients with mild and moderate clinical presentation.MethodsWe performed an epidemiological, observational, descriptive, and cross-sectional study carried out in Ecuador, with 1,447 participants between 18 and 66 years, non-hospitalized, with a molecular RT-PCR test for SARS-CoV2. We analyzed demographic characteristics according to sex, age group, clinical findings, behavior after diagnosis, family and social behavior, sequelae, clinical evolution, type of exposure, and personal history.ResultsThe sample analyzed had a mean age of 37 years (95% CI 18-66), women 713 individuals (49.27%), men 733 individuals (50.66%). Age group distribution was 18-30 years, 524 individuals (36.29%), 31-45, 538 individuals (37.26), and more of 45 years, 382 individuals (26.46%). 1416 individuals were mestizos (97.99%). According to the province of residence from Pichincha were 1019 patients (70.52%), followed by Imbabura, 93 patients (6.44%), and the others 335 (23.15%) patients come from all over the country. In women, the most common findings were fever >38°C (54.40%), sputum (27.43%) and hypoxia (16.32%); HTN (5.75%) and hypercholesterolemia (3.69%). Men were more prevalent in all other findings. Comorbidities were more prevalent in all those over 45 years of age. COVID-19 antibodies test was positive in 416 patients (28.85%). Neuropsychiatric symptoms such as sleep disorders, generalized anxiety disorder, depressed mood, and chronic fatigue were more prevalent in men than women. Still, generalized anxiety disorder and chronic fatigue were more common in individuals of 31 to 45 years. 868 patients (60.07%) were in contact with a known infected person, 318 patients (22.02%) were health workers, and 782 patients (57.63%) were informed about work exposure. 545 patients (37.72%) were overweight, primarily women 310 (42.29%). 609 patients (42.65%) showed symptoms after the acute period, and 331 individuals (23.49%) reported some sequelae.ConclusionThe epidemiological and clinical behavior of hospitalized and critical patients differs greatly from ambulatory or mild or moderate symptoms. It is essential to highlight those non-hospitalized patients constitute the predominant population of patients, hence the importance of adequate management that would directly affect the development of complicated forms and, consequently, the collapse of healthcare centers. It is vitally important to open more investigations that compare hospitalized and outpatient patients to have a clearer picture of the epidemic

Genetic Profile of the Kichwas (Quichuas) from Ecuador by Analysis of STR Loci

Allele frequency data for the STR systems CSF1PO, TPOX, TH01, D13S317, D16S539, D5S818, and D7S820 were determined in a population sample of unrelated, healthy Amerindian Kichwa individuals. All loci met Hardy–Weinberg expectations, and there was no evidence for association of alleles among the seven loci

A comprehensive study of Ecuadorian adult patients with a mild and moderate presentation of COViD-19

Aim To characterize non-hospitalized patients with mild and moderate clinical presentation. Methods We performed an epidemiological, observational, descriptive, and cross-sectional study carried out in Ecuador, with 1,447 participants between 18 and 66 years, non-hospitalized, with a molecular RT-PCR test for SARS-CoV2. We analyzed demographic characteristics according to sex, age group, clinical findings, behavior after diagnosis, family and social behavior, sequelae, clinical evolution, type of exposure, and personal history. Results The sample analyzed had a mean age of 37 years (95% CI 18–66), women 713 individuals (49.27%), men 733 individuals (50.66%). Age group distribution was 18–30 years, 524 individuals (36.29%), 31–45, 538 individuals (37.26), and more of 45 years, 382 individuals (26.46%). 1416 individuals were mestizos (97.99%). According to the province of residence from Pichincha were 1019 patients (70.52%), followed by Imbabura, 93 patients (6.44%), and the others 335 (23.15%) patients come from all over the country. In women, the most common findings were fever >38°C (54.40%), sputum (27.43%) and hypoxia (16.32%); HTN (5.75%) and hypercholesterolemia (3.69%). Men were more prevalent in all other findings. Comorbidities were more prevalent in all those over 45 years of age. COVID-19 antibodies test was positive in 416 patients (28.85%). Neuropsychiatric symptoms such as sleep disorders, generalized anxiety disorder, depressed mood, and chronic fatigue were more prevalent in men than women. Still, generalized anxiety disorder and chronic fatigue were more common in individuals of 31 to 45 years. 868 patients (60.07%) were in contact with a known infected person, 318 patients (22.02%) were health workers, and 782 patients (57.63%) were informed about work exposure. 545 patients (37.72%) were overweight, primarily women 310 (42.29%). 609 patients (42.65%) showed symptoms after the acute period, and 331 individuals (23.49%) reported some sequelae. Conclusion The epidemiological and clinical behavior of hospitalized and critical patients differs greatly from ambulatory or mild or moderate symptoms. It is essential to highlight those non-hospitalized patients constitute the predominant population of patients, hence the importance of adequate management that would directly affect the development of complicated forms and, consequently, the collapse of healthcare centers. It is vitally important to open more investigations that compare hospitalized and outpatient patients to have a clearer picture of the epidemic