Generación de un vector adenoviral para marcaje fluorescente selectivo de células tumorales

Introducción: Las células tumorales circulantes (CTC) son células que se originan de tumores primarios o de eventos metastásicos y que circulan libremente en el torrente sanguíneo de pacientes. Una de sus características más relevante es su capacidad ilimitada de división o inmortalidad. La expresión del gen de la transcriptasa reversa de telomerasa humana (hTERT), la enzima que mantiene la longitud de los telómeros, está desregulada en el 85-90% de las células tumorales. El uso de promotores tejido-específicos restringe la expresión génica o replicación viral a tejidos específicos. En este caso, el promotor de hTERT, altamente activo en la mayoría de las células tumorales, particularmente de cáncer colorrectal (CCR) como se mencionó previamente pero inactivo en células somáticas normales, resulta un candidato de promotor tejido-específico óptimo, por lo que su uso acoplado con proteínas fluorescentes mediante un vector adenoviral permitiría la detección selectiva de CTC. Objetivo: Generar un vector adenoviral diseñado para inducir la expresión selectiva de la proteína roja fluorescente (RFP) en células tumorales mediante la transducción con el gen RFP bajo el control del promotor hTERT. Materiales y métodos: Plásmidos: pUC57-hTERT-RFP, pShuttle, AdEasy-1. Cepas bacterianas: E. coli TOP10F’, BJ5183. Línea celular: HEK293. Se construyó el genoma del vector adenoviral a partir de la secuencia comercial hTERT-RFP clonada en pUC57. Dicha secuencia fue subclonada en el plásmido acarreador pShuttle. Luego éste se recombinó con el plásmido AdEasy. Se caracterizó y transfectó en células HEK293 para producir las partículas virales Ad-hTERTRFP. El vector adenoviral se caracterizó por inmunohistoquímica, PCR y microscopía de fluorescencia. Resultados: Se observó el efecto citopático característico de la infección por adenovirus en células HEK293 y mediante microscopía de fluorescencia se pudo observar la expresión de la RFP en su citoplasma. Conclusiones: Se diseñó, construyó, generó y caracterizó un vector adenoviral portador del promotor hTERT que induce la expresión de RFP en células que expresan altos niveles de telomerasa

Resección de Tumor Phyllodes con Reconstrucción Inmediata de la Mama, Cirugía Oncoplástica: Reporte de Caso y Revisión de la Literatura

Los tumores filoides son lesiones fibroepiteliales raros que constituyen solo entre el 0.3% y el 1% de todos los tumores primarios de mama (1). La mayoría de los casos se presentan entre los 35 y 55 años (2,3). La OMS estableció una clasificación histológica de los tumores filoides en benignos, limítrofes y malignos, basada en sus características histopatológicas. (3-6). El síntoma más común es la presencia de un bulto en la mama, generalmente localizado en el cuadrante superior externo de la mama y bilateral en solo el 1,8% de los casos. El tamaño varía entre 0,5 y 30 cm. (4,9,10) El tratamiento de los tumores filoides puede ser abordado de manera personalizada, tomando en cuenta el grado histopatológico y los márgenes quirúrgicos. Se ha observado que los tumores filoides limítrofes y malignos con márgenes quirúrgicos positivos o ≤1 mm presentan un mayor riesgo de recurrencia. En el caso de los tumores filoides benignos, un margen negativo estrecho puede ser óptimo, sin necesidad de cumplir con el margen de escisión tradicional de ≥10 mm (7,9). El papel de la radioterapia y la quimioterapia en el tratamiento de los tumores filoides sigue siendo incierto. (1,11,16,18). Es fundamental reconocer que la reconstrucción mamaria debe considerarse como un componente primordial en el proceso de tratamiento. Además, numerosos estudios respaldan que la reconstrucción no afecta negativamente la terapia adyuvante, ni interfiere con el tratamiento o el pronóstico de las pacientes (36,38). Se presenta caso de una paciente con tumor en mama derecha con reporte histológico de tumor filoides benigno sometida a reconstrucción inmediata con cirugía oncoplàstica con marcación prequirúrgica en patrón de Wise, cicatriz en T invertida con transposición del CAP un pedículo inferior con simetrización de mama contralateral, logrando una cirugía curativa con buen pronostico y evolución favorable.Phyllodes tumors are rare fibroepithelial lesions that constitute only between 0.3% and 1% of all primary breast tumors (1). Most cases occur between the ages of 35 and 55 (2,3). The WHO established a histological classification of phyllodes tumors into benign, borderline, and malignant, based on their histopathological characteristics (3-6). The most common symptom is the presence of a lump in the breast, usually located in the upper outer quadrant of the breast and bilateral in only 1.8% of cases. The size varies between 0.5 and 30 cm (4,9,10). The treatment of phyllodes tumors can be approached in a personalized manner, taking into account the histopathological grade and surgical margins. It has been observed that borderline and malignant phyllodes tumors with positive surgical margins or ≤1 mm present a higher risk of recurrence. In the case of benign phyllodes tumors, a narrow negative margin may be optimal, without the need to meet the traditional excision margin of ≥10 mm (7,9). The role of radiotherapy and chemotherapy in the treatment of phyllodes tumors remains uncertain (1,11,16,18). It is essential to recognize that breast reconstruction should be considered as a primary component in the treatment process. Furthermore, numerous studies support that reconstruction does not negatively affect adjuvant therapy, nor does it interfere with patient treatment or prognosis (36,38). We present a case of a patient with a tumor in the right breast with a histological report of benign phyllodes tumor undergoing immediate reconstruction with oncoplastic surgery with preoperative marking in Wise pattern, inverted T scar with transposition of the CAP an inferior pedicle with contralateral breast symmetrization, achieving curative surgery with good prognosis and favorable evolution

A922 Sequential measurement of 1 hour creatinine clearance (1-CRCL) in critically ill patients at risk of acute kidney injury (AKI)

Meeting abstrac

Implant Composed of Demineralized Bone and Mesenchymal Stem Cells Genetically Modified with AdBMP2/AdBMP7 for the Regeneration of Bone Fractures in Ovis aries

Adipose-derived mesenchymal stem cells (ADMSCs) are inducible to an osteogenic phenotype by the bone morphogenetic proteins (BMPs). This facilitates the generation of implants for bone tissue regeneration. This study evaluated the in vitro osteogenic differentiation of ADMSCs transduced individually and in combination with adenoviral vectors expressing BMP2 and BMP7. Moreover, the effectiveness of the implant containing ADMSCs transduced with the adenoviral vectors AdBMP2/AdBMP7 and embedded in demineralized bone matrix (DBM) was tested in a model of tibial fracture in sheep. This graft was compared to ewes implanted with untransduced ADMSCs embedded in the same matrix and with injured but untreated animals. In vivo results showed accelerated osteogenesis in the group treated with the AdBMP2/AdBMP7 transduced ADMSC graft, which also showed improved restoration of the normal bone morphology

Avances de las mujeres en las ciencias, las humanidades y todas las disciplinas. Libro científico II 2023

“Libro Científico II 2023”, es resultado del trabajo colaborativo entre pares de la División de Ciencias Biológicas y de la Salud (CBS) y la División de Ciencias Sociales y Humanidades (CSH)”Este libro contiene un conjunto de artículos científicos realizados en el contexto de la contingencia por COVID 19, desde los enfoques de las ciencias de la salud y las ciencias sociales. Su fin último es dar a conocer los avances científicos desde la perspectiva de género. De esta manera, la compilación incluye semblanzas de tres figuras relevantes de la academia mexicana: Ana Rosa López-Ferrari Aralia López González y Silvana Levi Levi con el propósito de sistematizar los aportes de cada una en los campos de la literatura, la botánica y la geografía respectivamente analizando de forma integral otros aspectos relevantes de lo que Bourdieu llamaría sus condiciones sociales de posibilidad. Por otro lado los artículos que conforman el libro se enfocan en: El estudio de posibles causas de la obesidad; exploraron los efectos psicológicos del confinamiento provocado por la pandemia por COVID 19 sobre los hábitos alimenticios y del sueño respectivamente; la experiencia exitosa de transformación cultivo de hortalizas de forma orgánica y sustentable durante un proceso de sobrevivencia y adaptación a las duras condiciones económicas y sociales producidas por la pandemia por COVID-19; La investigación con resultados alentadores, las características de sustancias que tentativamente puedan reemplazar aquellas con las que se trata el glioblastoma, de una manera más eficiente y con menos efectos colaterales; El incremento de los conflictos domésticos, la violencia por razones de género y feminicidios durante la pandemia; La valoración de los marcos de competencias digitales en España para diferentes áreas académicas en las que se implementó la educación a distancia con motivo de la pandemia por COVID-19.Yadira Alatriste Martínez, editora, compiladora y directora del equipo editorial; César Ulises Hernández Franco; Brandon Antonio Navarrete Rubio; Claudia Angélica Chávez Gutiérrez; Zaina Yeridni Martínez Leyva; Abraham Hernández Villegas; Ingrid Karina Mayen Delgado; Juana Aidet Jiménez Velasco, diseño y maquetación. Andrea Forero Castillo, prólogo; Amalia Patricia Gaytán Sánchez, introducción

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Burden of disease scenarios for 204 countries and territories, 2022–2050: a forecasting analysis for the Global Burden of Disease Study 2021

Background: Future trends in disease burden and drivers of health are of great interest to policy makers and the public at large. This information can be used for policy and long-term health investment, planning, and prioritisation. We have expanded and improved upon previous forecasts produced as part of the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) and provide a reference forecast (the most likely future), and alternative scenarios assessing disease burden trajectories if selected sets of risk factors were eliminated from current levels by 2050. Methods: Using forecasts of major drivers of health such as the Socio-demographic Index (SDI; a composite measure of lag-distributed income per capita, mean years of education, and total fertility under 25 years of age) and the full set of risk factor exposures captured by GBD, we provide cause-specific forecasts of mortality, years of life lost (YLLs), years lived with disability (YLDs), and disability-adjusted life-years (DALYs) by age and sex from 2022 to 2050 for 204 countries and territories, 21 GBD regions, seven super-regions, and the world. All analyses were done at the cause-specific level so that only risk factors deemed causal by the GBD comparative risk assessment influenced future trajectories of mortality for each disease. Cause-specific mortality was modelled using mixed-effects models with SDI and time as the main covariates, and the combined impact of causal risk factors as an offset in the model. At the all-cause mortality level, we captured unexplained variation by modelling residuals with an autoregressive integrated moving average model with drift attenuation. These all-cause forecasts constrained the cause-specific forecasts at successively deeper levels of the GBD cause hierarchy using cascading mortality models, thus ensuring a robust estimate of cause-specific mortality. For non-fatal measures (eg, low back pain), incidence and prevalence were forecasted from mixed-effects models with SDI as the main covariate, and YLDs were computed from the resulting prevalence forecasts and average disability weights from GBD. Alternative future scenarios were constructed by replacing appropriate reference trajectories for risk factors with hypothetical trajectories of gradual elimination of risk factor exposure from current levels to 2050. The scenarios were constructed from various sets of risk factors: environmental risks (Safer Environment scenario), risks associated with communicable, maternal, neonatal, and nutritional diseases (CMNNs; Improved Childhood Nutrition and Vaccination scenario), risks associated with major non-communicable diseases (NCDs; Improved Behavioural and Metabolic Risks scenario), and the combined effects of these three scenarios. Using the Shared Socioeconomic Pathways climate scenarios SSP2-4.5 as reference and SSP1-1.9 as an optimistic alternative in the Safer Environment scenario, we accounted for climate change impact on health by using the most recent Intergovernmental Panel on Climate Change temperature forecasts and published trajectories of ambient air pollution for the same two scenarios. Life expectancy and healthy life expectancy were computed using standard methods. The forecasting framework includes computing the age-sex-specific future population for each location and separately for each scenario. 95% uncertainty intervals (UIs) for each individual future estimate were derived from the 2·5th and 97·5th percentiles of distributions generated from propagating 500 draws through the multistage computational pipeline. Findings: In the reference scenario forecast, global and super-regional life expectancy increased from 2022 to 2050, but improvement was at a slower pace than in the three decades preceding the COVID-19 pandemic (beginning in 2020). Gains in future life expectancy were forecasted to be greatest in super-regions with comparatively low life expectancies (such as sub-Saharan Africa) compared with super-regions with higher life expectancies (such as the high-income super-region), leading to a trend towards convergence in life expectancy across locations between now and 2050. At the super-region level, forecasted healthy life expectancy patterns were similar to those of life expectancies. Forecasts for the reference scenario found that health will improve in the coming decades, with all-cause age-standardised DALY rates decreasing in every GBD super-region. The total DALY burden measured in counts, however, will increase in every super-region, largely a function of population ageing and growth. We also forecasted that both DALY counts and age-standardised DALY rates will continue to shift from CMNNs to NCDs, with the most pronounced shifts occurring in sub-Saharan Africa (60·1% [95% UI 56·8–63·1] of DALYs were from CMNNs in 2022 compared with 35·8% [31·0–45·0] in 2050) and south Asia (31·7% [29·2–34·1] to 15·5% [13·7–17·5]). This shift is reflected in the leading global causes of DALYs, with the top four causes in 2050 being ischaemic heart disease, stroke, diabetes, and chronic obstructive pulmonary disease, compared with 2022, with ischaemic heart disease, neonatal disorders, stroke, and lower respiratory infections at the top. The global proportion of DALYs due to YLDs likewise increased from 33·8% (27·4–40·3) to 41·1% (33·9–48·1) from 2022 to 2050, demonstrating an important shift in overall disease burden towards morbidity and away from premature death. The largest shift of this kind was forecasted for sub-Saharan Africa, from 20·1% (15·6–25·3) of DALYs due to YLDs in 2022 to 35·6% (26·5–43·0) in 2050. In the assessment of alternative future scenarios, the combined effects of the scenarios (Safer Environment, Improved Childhood Nutrition and Vaccination, and Improved Behavioural and Metabolic Risks scenarios) demonstrated an important decrease in the global burden of DALYs in 2050 of 15·4% (13·5–17·5) compared with the reference scenario, with decreases across super-regions ranging from 10·4% (9·7–11·3) in the high-income super-region to 23·9% (20·7–27·3) in north Africa and the Middle East. The Safer Environment scenario had its largest decrease in sub-Saharan Africa (5·2% [3·5–6·8]), the Improved Behavioural and Metabolic Risks scenario in north Africa and the Middle East (23·2% [20·2–26·5]), and the Improved Nutrition and Vaccination scenario in sub-Saharan Africa (2·0% [–0·6 to 3·6]). Interpretation: Globally, life expectancy and age-standardised disease burden were forecasted to improve between 2022 and 2050, with the majority of the burden continuing to shift from CMNNs to NCDs. That said, continued progress on reducing the CMNN disease burden will be dependent on maintaining investment in and policy emphasis on CMNN disease prevention and treatment. Mostly due to growth and ageing of populations, the number of deaths and DALYs due to all causes combined will generally increase. By constructing alternative future scenarios wherein certain risk exposures are eliminated by 2050, we have shown that opportunities exist to substantially improve health outcomes in the future through concerted efforts to prevent exposure to well established risk factors and to expand access to key health interventions

Global age-sex-specific mortality, life expectancy, and population estimates in 204 countries and territories and 811 subnational locations, 1950–2021, and the impact of the COVID-19 pandemic: a comprehensive demographic analysis for the Global Burden of Disease Study 2021

Background: Estimates of demographic metrics are crucial to assess levels and trends of population health outcomes. The profound impact of the COVID-19 pandemic on populations worldwide has underscored the need for timely estimates to understand this unprecedented event within the context of long-term population health trends. The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2021 provides new demographic estimates for 204 countries and territories and 811 additional subnational locations from 1950 to 2021, with a particular emphasis on changes in mortality and life expectancy that occurred during the 2020–21 COVID-19 pandemic period. Methods: 22 223 data sources from vital registration, sample registration, surveys, censuses, and other sources were used to estimate mortality, with a subset of these sources used exclusively to estimate excess mortality due to the COVID-19 pandemic. 2026 data sources were used for population estimation. Additional sources were used to estimate migration; the effects of the HIV epidemic; and demographic discontinuities due to conflicts, famines, natural disasters, and pandemics, which are used as inputs for estimating mortality and population. Spatiotemporal Gaussian process regression (ST-GPR) was used to generate under-5 mortality rates, which synthesised 30 763 location-years of vital registration and sample registration data, 1365 surveys and censuses, and 80 other sources. ST-GPR was also used to estimate adult mortality (between ages 15 and 59 years) based on information from 31 642 location-years of vital registration and sample registration data, 355 surveys and censuses, and 24 other sources. Estimates of child and adult mortality rates were then used to generate life tables with a relational model life table system. For countries with large HIV epidemics, life tables were adjusted using independent estimates of HIV-specific mortality generated via an epidemiological analysis of HIV prevalence surveys, antenatal clinic serosurveillance, and other data sources. Excess mortality due to the COVID-19 pandemic in 2020 and 2021 was determined by subtracting observed all-cause mortality (adjusted for late registration and mortality anomalies) from the mortality expected in the absence of the pandemic. Expected mortality was calculated based on historical trends using an ensemble of models. In location-years where all-cause mortality data were unavailable, we estimated excess mortality rates using a regression model with covariates pertaining to the pandemic. Population size was computed using a Bayesian hierarchical cohort component model. Life expectancy was calculated using age-specific mortality rates and standard demographic methods. Uncertainty intervals (UIs) were calculated for every metric using the 25th and 975th ordered values from a 1000-draw posterior distribution. Findings: Global all-cause mortality followed two distinct patterns over the study period: age-standardised mortality rates declined between 1950 and 2019 (a 62·8% [95% UI 60·5–65·1] decline), and increased during the COVID-19 pandemic period (2020–21; 5·1% [0·9–9·6] increase). In contrast with the overall reverse in mortality trends during the pandemic period, child mortality continued to decline, with 4·66 million (3·98–5·50) global deaths in children younger than 5 years in 2021 compared with 5·21 million (4·50–6·01) in 2019. An estimated 131 million (126–137) people died globally from all causes in 2020 and 2021 combined, of which 15·9 million (14·7–17·2) were due to the COVID-19 pandemic (measured by excess mortality, which includes deaths directly due to SARS-CoV-2 infection and those indirectly due to other social, economic, or behavioural changes associated with the pandemic). Excess mortality rates exceeded 150 deaths per 100 000 population during at least one year of the pandemic in 80 countries and territories, whereas 20 nations had a negative excess mortality rate in 2020 or 2021, indicating that all-cause mortality in these countries was lower during the pandemic than expected based on historical trends. Between 1950 and 2021, global life expectancy at birth increased by 22·7 years (20·8–24·8), from 49·0 years (46·7–51·3) to 71·7 years (70·9–72·5). Global life expectancy at birth declined by 1·6 years (1·0–2·2) between 2019 and 2021, reversing historical trends. An increase in life expectancy was only observed in 32 (15·7%) of 204 countries and territories between 2019 and 2021. The global population reached 7·89 billion (7·67–8·13) people in 2021, by which time 56 of 204 countries and territories had peaked and subsequently populations have declined. The largest proportion of population growth between 2020 and 2021 was in sub-Saharan Africa (39·5% [28·4–52·7]) and south Asia (26·3% [9·0–44·7]). From 2000 to 2021, the ratio of the population aged 65 years and older to the population aged younger than 15 years increased in 188 (92·2%) of 204 nations. Interpretation: Global adult mortality rates markedly increased during the COVID-19 pandemic in 2020 and 2021, reversing past decreasing trends, while child mortality rates continued to decline, albeit more slowly than in earlier years. Although COVID-19 had a substantial impact on many demographic indicators during the first 2 years of the pandemic, overall global health progress over the 72 years evaluated has been profound, with considerable improvements in mortality and life expectancy. Additionally, we observed a deceleration of global population growth since 2017, despite steady or increasing growth in lower-income countries, combined with a continued global shift of population age structures towards older ages. These demographic changes will likely present future challenges to health systems, economies, and societies. The comprehensive demographic estimates reported here will enable researchers, policy makers, health practitioners, and other key stakeholders to better understand and address the profound changes that have occurred in the global health landscape following the first 2 years of the COVID-19 pandemic, and longer-term trends beyond the pandemic