212 research outputs found
"Efecto in vitro de Escherichia coli uropatógena en la calidad espermática humana."
El espermatozoide es una célula altamente especializada encargada de
llevar el material genético paterno hasta el tracto reproductivo femenino en
búsqueda del oocito, sin embargo durante su desplazamiento puede
interactuar con microorganismos que puede transportar desencadenando
procesos infecciosos que alteran el éxito reproductivo. El objetivo de este
estudio fue determinar el efecto in vitro de Escherichia coli uropatógena en
el parámetro de motilidad, vitalidad, aglutinación y el impacto en la
fragmentación de ADN espermático humano. Se evaluaron 40 muestras de
semen provenientes de jóvenes con edades comprendidas entre 20 y 25
años. Se obtuvo la cepa microbiana ATCC E.coli 25922 por medio de
compra marca Microbiologics®, y se evaluó el efecto de E.coli en la calidad
espermática en humano, se incubó 1ml de semen con buffer Flushing
(Origio®) con la cepa de E. coli, se utilizó cuatro concentraciones
diferentes, para obtener una relación de E. coli a espermatozoides de
0,5:9,5 1:9, 3:7, y 5:5. Como resultado se observa que disminuye los
valores de vitalidad (t1) a 77,36% +/-4,50, el segundo tratamiento (t2)
disminuye a 72,93% +/-5,29, el tercer tratamiento (t3) a 67,84% +/-7,16, y
el cuarto tratamiento (t4) a 59,35% +/-5,43.Motilidad (t1) 42,69% +/-27,66,
el segundo tratamiento 37,77% +/-29,51, el tercer tratamiento (t3) 26,27%
+/-35,89 y el cuarto tratamiento (t4) disminuye a 17,79% +/-24,61. Es
evidente que la presencia de la cepa microbiana ATCC E.coli 25922 causa
un efecto patógeno de aglutinación de espermatozoides en todos los
tratamientos (p<0,05) en el primer tratamiento (t1) 60,02% +/-25,96, el
segundo tratamiento (t2) 65,03% +/- 24,15, en el tercer tratamiento (t3)
68,61% +/-22,29 y el cuarto tratamiento (t4) 72,95% +/-21,08, se demuestra
que a medida que aumenta la concentración de E.coli aumenta la
fragmentación de ADN espermático.
The spermatozoon is a highly specialized cell responsible for carrying the
paternal genetic material to the female reproductive tract in search of the
oocyte, however during its displacement it can interact with microorganisms
that can transport triggering infectious processes that alter reproductive
success. The objective of this study was to determine the in vitro effect of
uropathogenic Escherichia coli on the motility, vitality, agglutination
parameter and the impact on fragmentation of human sperm DNA. We
evaluated 40 semen samples from young people aged between 20 and 25
years. The ATCC E.coli 25922 microbial strain was obtained through the
purchase of Microbiologics® brand, and the effect of E. coli on sperm quality
in human was evaluated, 1ml of semen was incubated with Flushing buffer
(Origio®) with the strain of E. coli, four different concentrations were used,
to obtain a ratio of E. coli to spermatozoa of 0.5: 9.5 1: 9, 3: 7, and 5: 5. As
a result it is observed that it diminishes the values of vitality (t1) to 77.36%
+/- 4.50, the second treatment (t2) decreases to 72.93% +/- 5.29, the third
treatment (t3) to 67.84% +/- 7.16, and the fourth treatment (t4) to 59.35%
+/- 5,43.Motility (t1) 42.69% +/- 27.66, the second treatment 37 , 77% +/-
29.51, the third treatment (t3) 26.27% +/- 35.89 and the fourth treatment (t4)
decreases to 17.79% +/- 24.61. It is evident that the presence of the ATCC
E.coli 25922 microbial strain causes a pathogenic effect of sperm
agglutination in all treatments (p <0.05) in the first treatment (t1) 60.02% +/-
25.96, the second treatment (t2) 65.03% +/- 24.15, in the third treatment (t3)
68.61% +/- 22.29 and the fourth treatment (t4) 72.95% +/- 21.08, it is
demonstrated that as the concentration of E. coli increases, the
fragmentation of sperm DNA increases.Tesi
Metal-Organic Nanocapsules with Functionalized s-Heptazine Ligands
A metalloorganic capsule was synthesized where the ligand is a derivative of heptazine with three carboxylic groups that are coordinated to CuII cations, forming paddle-wheel motifs. Each nanocapsule is neutral, with 12 CuII centers and 8 ligands adopting a rhombicuboctahedron shape. It has almost 3 nm diameter, and the main intermolecular interactions in the solid are π··· π stacking between the C6N7 heptazine moieties. The nanocapsules can form monolayers deposited on graphite as observed by atomic force microscopy, which confirms their stability in solution
Hazard assessment of different-sized polystyrene nanoplastics in hematopoietic human cell lines
Altres ajuts: acords transformatius de la UABThe environmental presence of micro/nanoplastics (MNPLs) is an environmental and human health concern. Such MNPLs can result from the physicochemical/biological degradation of plastic goods (secondary MNPLs) or can result from industrial production at that size, for different commercial purposes (primary MNPLs). Independently of their origin, the toxicological profile of MNPLs can be modulated by their size, as well as by the ability of cells/organisms to internalize them. To get more information on these topics we have determined the ability of three different sizes of polystyrene MNPLs (50, 200, and 500 nm) to produce different biological effects in three different human hematopoietic cell lines (Raji-B, THP-1, and TK6). Results show that none of the three sizes was able to induce toxicity (growth ability) in any of the tested cell types. Although transmission electron microscopy and confocal images showed cell internalization in all the cases, their quantification by flow cytometry demonstrated an important uptake by Raji-B and THP-1 cells, in comparison with TK6 cells. For the first ones, the uptake was negatively associated with the size. Interestingly, when the loss of mitochondrial membrane potential was determined, dose-related effects were observed for Raji-B and THP-1 cells, but not for TK6 cells. These effects were observed for the three different sizes. Finally, when oxidative stress induction was evaluated, no clear effects were observed for the different tested combinations. Our conclusion is that size, biological endpoint, and cell type are aspects modulating the toxicological profile of MNPLs
Experimental Models of Liquid Biopsy in Hepatocellular Carcinoma Reveal Clone-Dependent Release of Circulating Tumor DNA.
Liquid biopsy, the molecular analysis of tumor components released into the bloodstream, has emerged as a noninvasive and resourceful means to access genomic information from cancers. Most data derived from translational studies showcase its numerous potential clinical applications. However, data from experimental models are scarce, and little is known about the underlying mechanisms and factors controlling the release of circulating tumor DNA (ctDNA) and cells (CTCs). This study aimed to model liquid biopsy in hepatocellular carcinoma xenografts and to study the dynamics of release of ctDNA and CTCs; this included models of intratumoral heterogeneity (ITH) and metastatic disease. We quantified ctDNA by quantitative polymerase chain reaction (PCR) targeting human long interspersed nuclear element group 1; targeted mutation analysis was performed with digital droplet PCR. CTCs were traced by flow cytometry. Results demonstrated the feasibility of detecting ctDNA, including clone-specific mutations, as well as CTCs in blood samples of mice. In addition, the concentration of ctDNA and presence of tumor-specific mutations reflected tumor progression, and detection of CTCs was associated with metastases. Our ITH model suggested differences in the release of DNA fragments impacted by the cell-clone origin and the treatment. Conclusion: These data present new models to study liquid biopsy and its underlying mechanisms and highlighted a clone-dependent release of ctDNA into the bloodstream
Cortometraje “Dante”
“Dante” es un proyecto que invita a nuestra generación a reflexionar sobre la presencia de las
redes sociales en nuestras vidas y las repercusiones que acarrea el uso de las mismas en
nuestro estado emocional. Así, el proyecto es creado con el objetivo de proponer una nueva
forma de interpretar nuestra relación con las redes sociales mediante la representación de
símiles del mundo digital y visual. No se presenta a un individuo que consume los contenidos
de manera pasiva desde el otro lado de la pantalla de su dispositivo, sino que más bien se
propone la existencia de un ser sin sexo definido, inmerso en una representación del mundo
de las redes, el cual experimenta de manera directa. Además, a través de la secuencia del
recorrido del protagonista, se busca que el corto sea una ventana para el espectador hacia su
mundo interno, el cual se encuentra en constante contacto con el mundo digital y con las
emociones que pueden experimentarse por la inmersión en las redes. Finalmente, esperamos
que los espectadores vean en “Dante” una representación de esos momentos en los que nos
hemos sentido abrumados por lo que encontramos al navegar en las redes, lo que incluso
puede consumirnos sin darnos cuenta."Dante" is a project that invites our generation to reflect on the presence of social networks in
our lives and the repercussions that the use of them has on our emotional state. Thus, the
project is created with the aim of proposing a new way of interpreting our relationship with
social networks, through the representation of similes of the digital and visual world. An
individual who consumes content passively from the other side of the screen of his device is
not presented in this short, but rather the existence of a being without a defined sex is
proposed, who is immersed in a representation of the world of social networks, which he
experiences directly. In addition, through the sequence of the protagonist's journey, the short
film is intended to be a window for the viewer to his internal world, which is in constant
contact with the digital world and with the emotions that can be experienced by getting into
social networks. Finally, we hope that viewers see in "Dante", a representation of those
moments in which we have been overwhelmed by what we find when browsing the Internet,
which could consume us without even realizing.Trabajo de investigació
Chemical composition of rainwater under two events of aerosol transport: A Saharan dust outbreak and wildfires
Modelos alométricos para estimar altura de árvores em florestas ecotonais do norte da Amazônia
Allometric models defining the relationship between stem diameter and total tree height in the Amazon basin are important because they refine the estimates of tree carbon stocks and flow in the region. This study tests different allometric models to estimate the total tree height from the stem diameter in an ecotone zone between ombrophilous and seasonal forests in the Brazilian state of Roraima, in northern Amazonia. Stem diameter and total height were measured directly in 65 recently fallen trees (live or dead). Linear and nonlinear regressions were tested to represent the D:H relation in this specific ecotone zone. Criteria for model selection were the standard error of the estimate (Syx) and the adjusted coefficient of determination (R²adj), complemented by the Akaike Information Criterion (AIC). Analysis of residuals of the most parsimonious nonlinear models showed a tendency to overestimate the total tree height for trees in the 20-40 cm diameter range. Application of our best fitted model (Michaelis-Menten) indicated that previously published general equations for the tropics that use diameter as the independent variable can either overestimate tree height in the study area by 10-29% (Weibull models) or underestimate it by 8% (climate-based models). We concluded that our site-specific model can be used in the ecotone forests studied in Roraima because it realistically reflects the local biometric relationships between stem diameter and total tree height. Studies need to be expanded in peripheral areas of northern Amazonia in order to reduce uncertainties in biomass and carbon estimates that use the tree height as a variable in general models.Modelos alométricos que definem o relacionamento entre diâmetro do tronco e a altura total da árvore na bacia amazônica são importantes porque refinam as estimativas de fluxo e estoques de carbono arbóreo na região. Este estudo testou diferentes modelos alométricos para estimar a altura total de árvores a partir do diâmetro do tronco em uma zona de ecótono entre florestas ombrófilas e sazonais no estado de Roraima, norte da Amazônia. Diâmetro do tronco e altura total foram medidos de forma direta em 65 árvores tombadas recentemente (vivas e mortas). Regressões linear e não-linear foram testadas para representar a relação D:H nesta zona específica de ecótono. Os critérios de seleção dos modelos foram o erro padrão da estimativa (Syx), o coeficiente de determinação ajustado (R²adj) e o Critério de Informação de Akaike (AIC). A análise dos resíduos dos modelos não-lineares mais parcimoniosos mostrou uma tendência de superestimar a altura total para árvores entre 20-40 cm de diâmetro do tronco. A aplicação do modelo melhor ajustado (Michaelis-Menten) indicou que equações gerais publicadas previamente para os trópicos que usam diâmetro como variável independente podem superestimar em 10-29% (modelos Weibull) ou subestimar em 8% (modelos baseados no clima) a altura das árvores na área de estudo. Nós concluímos que o modelo de melhor ajuste pode ser usado nas florestas ecotonais estudadas em Roraima, porque ele reflete realisticamente o relacionamento biométrico local entre diâmetro do tronco e altura total da árvore. É necessário expandir os estudos para outras áreas periféricas do norte da Amazônia, com o intuito de reduzir as incertezas em estimativas de biomassa e carbono arbóreo que adotem altura da árvore como uma variável em modelos gerais
CRISPR interference interrogation of COPD GWAS genes reveals the functional significance of desmoplakin in iPSC-derived alveolar epithelial cells
Genome-wide association studies (GWAS) have identified dozens of loci associated with chronic obstructive pulmonary disease (COPD) susceptibility; however, the function of associated genes in the cell type(s) affected in disease remains poorly understood, partly due to a lack of cell models that recapitulate human alveolar biology. Here, we apply CRISPR interference to interrogate the function of nine genes implicated in COPD by GWAS in induced pluripotent stem cell–derived type 2 alveolar epithelial cells (iAT2s). We find that multiple genes implicated by GWAS affect iAT2 function, including differentiation potential, maturation, and/or proliferation. Detailed characterization of the GWAS gene DSP demonstrates that it regulates iAT2 cell-cell junctions, proliferation, mitochondrial function, and response to cigarette smoke–induced injury. Our approach thus elucidates the biological function, as well as disease-relevant consequences of dysfunction, of genes implicated in COPD by GWAS in type 2 alveolar epithelial cells.This work was supported by a CJ Martin Early Career Fellowship from the Australian National Health and Medical Research Council awarded to R.B.W.; NIH grant F30HL147426 awarded to K.M.A.; NIH grants U01TR001810, R01DK101501, and R01DK117940 awarded to A.A.W.; NIH grants R01HL135142, R01HL137927, and R01HL147148 awarded to M.H.C.; and NIH grants R01HL127200 and R01HL148667 awarded to X.Z
Human iPSC-hepatocyte modeling of alpha-1 antitrypsin heterozygosity reveals metabolic dysregulation and cellular heterogeneity
Individuals homozygous for the “Z” mutation in alpha-1 antitrypsin deficiency are known to be at increased risk for liver disease. It has also become clear that some degree of risk is similarly conferred by the heterozygous state. A lack of model systems that recapitulate heterozygosity in human hepatocytes has limited the ability to study the impact of a single Z alpha-1 antitrypsin (ZAAT) allele on hepatocyte biology. Here, we describe the derivation of syngeneic induced pluripotent stem cells (iPSCs) engineered to determine the effects of ZAAT heterozygosity in iPSC-hepatocytes (iHeps). We find that heterozygous MZ iHeps exhibit an intermediate disease phenotype and share with ZZ iHeps alterations in AAT protein processing and downstream perturbations including altered endoplasmic reticulum (ER) and mitochondrial morphology, reduced mitochondrial respiration, and branch-specific activation of the unfolded protein response in cell subpopulations. Our model of MZ heterozygosity thus provides evidence that a single Z allele is sufficient to disrupt hepatocyte homeostatic function.This work was supported by an Alpha-1 Foundation John W. Walsh Translational Research Award (to J.E.K.); a CJ Martin Early Career Fellowship from the Australian National Health and Medical Research Council (to R.B.W.); NIH grant R01HL095993 (to D.N.K.); and NIH grants R01DK101501 (to A.A.W.) and R01DK117940 (to A.N.H. and A.A.W.). iPSC distribution and disease modeling is supported by NIH grants U01TR001810 (to D.N.K. and A.A.W.) and N0175N92020C00005 (to D.N.K.); and by The Alpha-1 Project (TAP), a wholly owned subsidiary of the Alpha-1 Foundation (to D.N.K. and A.A.W.)
High-density single cell mRNA sequencing to characterize circulating tumor cells in hepatocellular carcinoma.
Patients with hepatocellular carcinoma (HCC) release tumor cells to the bloodstream, which can be detected using cell surface markers. Despite numerous reports suggest a direct correlation between the number of circulating tumor cells (CTCs) and poor clinical outcomes, few studies have provided a thorough molecular characterization of CTCs. Due to the limited access to tissue samples in patients at advanced stages of HCC, it is crucial to develop new technologies to identify HCC cancer drivers in routine clinical conditions. Here, we describe a method that sequentially combines image flow cytometry and high density single-cell mRNA sequencing to identify CTCs in HCC patients. Genome wide expression profiling of CTCs using this approach demonstrates CTC heterogeneity and helps detect known oncogenic drivers in HCC such as IGF2. This integrated approach provides a novel tool for biomarker development in HCC using liquid biopsy
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