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Micronuclei and nuclear anomalies in Mexico’s indigenous population

BLANCA PATRICIA LAZALDE RAMOS (2017)

Objective. To determine the number of micronuclei and nuclear anomalies in Mexico’s indigenous population. Materials and methods. One hundred twenty indigenous individuals were evaluated, including thirty from the ethnicities Cora, Huichol, Tarahumara and Tepehuano. The number of micronuclei (MN) and any nuclear abnormality (NA) in oral mucosa cells, including cells with nuclear buds, binucleated cells, cells with karyolysis, karyorrhetic, condensed chromatin and pyknotic cells were determined for each participant. Results. Tepehuano and Tarahumaras showed the greatest damage to DNA. The Tepehuano group presented the highest number of MN and NA, this being a significant difference (p < 0.05) compared with the rest of the studied groups. This group also presented the highest herbicide exposure (46.7%). In relation to the smoking and drinking habits, these were more frequent in the Tarahumara group (33.3 and 50% respectively). Conclusion. The ethnic diversity, habits and customs may influence the DNA nuclear integrity in the Amerindian groups.

Producción Científica de la Universidad Autónoma de Zacatecas UAZ

Article

BIOLOGÍA Y QUÍMICA micronuclei nuclear abnormalities indigenous population DNA

Sincronización de células de tabaco (Nicotiana tabacum) línea celular NT-1

León Francisco Ruiz Herrera (2007)

Instituto de Investigaciones Químico Biológicas. Maestría en Ciencias en Biología Experimental

The cell cycle is the period that elapses from one division to the next. Each Cell is the product of a cell cycle, which comprises the ordered sequence of the Events G1→G2→M (Figure 1.1), where a eukaryotic cell doubles its Chromosomes and divides into two cells. An overview of the cell cycle allows Observe how the cell passes through two major stages: cell division, or phase M, and The interface (comprising G1, S and G2 phases) (Buchanan et al., 2000, Morgan, 2007). The interface, which elapses between two cell divisions, is the most Of the cell cycle. During phase G1 (G gap) materials are synthesized as RNA and proteins; In the S phase or of synthesis the DNA replication takes place, besides Synthesize some proteins such as histones. After replication is complete, the cell Enters the phase G2 where compounds necessary for the division are synthesized and begins Gradual condensation of the chromatin, which is completed in the early stages of Mitosis giving rise to chromosomes visible under the microscope, showing its typical appearance Of two chromatids and four arms (Buchanan et al., 2000, Dashek and Harrison, 2006; Morgan, 2007). Phase M is the shortest period of the cell cycle, during which the Mitosis (or karyokinesis), ie nuclear division and cytokinesis, and the division of the Cell (which has previously duplicated its genetic and cytoplasmic material) in two Identical daughter cells.

El ciclo celular es el periodo que transcurre de una división a la siguiente. Cada célula es el producto de un ciclo celular, el cual comprende la secuencia ordenada de los eventos G1→G2→M (Figura 1.1), donde una célula eucarionte duplica sus cromosomas y se divide en dos células. Una visión global del ciclo celular permite observar cómo la célula transcurre por dos grandes etapas: la división celular, o fase M, y la interfase (que comprende las fases G1, S y G2 ) (Buchanan et al., 2000; Morgan, 2007). La interfase, tiempo que transcurre entre dos divisiones celulares, es el lapso más prolongado del ciclo celular. Durante la fase G1 (G de gap) se sintetizan materiales, como RNA y proteínas; en la fase S o de síntesis se produce la replicación del DNA, además se sintetizan algunas proteínas como las histonas. Una vez finalizada la replicación, la célula entra en la fase G2 donde se sintetizan compuestos necesarios para la división y se inicia la condensación gradual de la cromatina, que se completa en las primeras etapas de la mitosis dando lugar a cromosomas visibles al microscopio, mostrando su aspecto típico de dos cromátidas y cuatro brazos (Buchanan et al., 2000; Dashek and Harrison; 2006; Morgan, 2007). La fase M es el periodo más corto del ciclo celular, durante el cual tiene lugar la mitosis (o cariocinesis), es decir la división nuclear y la citocinesis, y la división de la célula (que previamente ha duplicado su material genético y citoplasmático) en dos células hijas idénticas.

Master thesis

CIENCIAS AGROPECUARIAS Y BIOTECNOLOGÍA IIQB-M-2007-0002 Células Tabaco DNA

Participación de los productos de los genes gpa11 y gpa12 en la esporulación y germinación de Mucor circinelloides

Nancy Yadira Reyes Mares (2017)

Instituto de Investigaciones Químico Biológicas. Maestría en Ciencias en Biología Experimental

The heterotrimeric G proteins are made up of the Gα, Gβ and Gγ protein subunits; In fungi these proteins are involved in different biological processes such as growth, differentiation, pathogenesis, among others. In the Mucor circinelloides fungus in a total of 12 Gα, 3 Gβ and 3 Gγ subunits, this being the largest repertoire of heterotrimeric G subunits in the fungus kingdom; however, its biological function is still unknown. The analysis of the mRNA levels of the 12 Gα genes (gpa1-12) during the dimorphism of M. circinelloides revealed that the transcripts of the gpa11 and gpa12 genes were overexpressed in the spore stage with respect to the rest of the Gα genes, Suggesting a possible participation in this morphological stage. The objective of this work was to demonstrate the participation of the products of the genes gpa11 and gpa12 in the sporulation and / or growth of M. circinelloides. Phenotypic analysis in terms of the growth and germination of spores of the single mutants in the Δgpa11 and Δgpa12 genes did not show significant differences with respect to the wild strain Mu402; However, a 30% reduction in spore production was observed for both mutant strains relative to the wild strain. In the double mutant strain Δgpa11 / Δgpa12 the spore production was similar to the simple mutants, they showed a significant difference in size of, 15 μm in average, whereas the spores of the simple and wild mutants had a size of 10 Μm on average. On the other hand there was a significant decrease in the biomass obtained in liquid cultures, compared to the other strains. In addition, the size of the hyphae of the double mutants were considerably longer, compared to the other strains. On the other hand, they analyzed the transcript levels of s6k genes (white kinase of the protein kinase TOR) and cnaA (catalytic subunit of calceneurin), proteins involved in cell differentiation processes, among others.

Las proteínas G heterotriméricas están conformadas por las subunidades proteicas Gα, Gβ y Gγ; En los hongos estas proteínas están involucradas en diferentes procesos biológicos tales como el crecimiento, diferenciación, patogénesis, entre otros. En el hongo Mucor circinelloides se han encontrado un total de 12 subunidades Gα, 3 Gβ y 3 Gγ, siendo este el mayor repertorio de subunidades G heterotriméricas en el reino fungí; sin embargo, aún se desconoce su función biológica. El análisis de los niveles de RNAm de los 12 genes Gα (gpa1-12) durante el dimorfismo de M. circinelloides, reveló que los transcritos de los genes gpa11 y gpa12 fueron sobreexpresados en el estadio de espora respecto al resto de los genes Gα, sugiriendo una posible participación en este estadio morfológico. El objetivo de este trabajo consistió en demostrar la participación de los productos de los genes gpa11 y gpa12 en la esporulación y/o crecimiento de M. circinelloides. El análisis fenotípico en términos del crecimiento y germinación de las esporas de las mutantes simples en los genes Δgpa11 y Δgpa12 no mostró diferencias significativas con respecto a la cepa silvestre Mu402; sin embargo, se observó un 30% de disminución en la producción de esporas para ambas cepas mutantes respecto a la cepa silvestre. En la cepa doble mutante Δgpa11/Δgpa12 la producción de esporas fue similar a las mutantes simples, éstas presentaron una diferencia significativa en el tamaño de, 15 μm en promedio, mientras que las esporas de las mutantes simples y silvestre, tuvieron un tamaño de 10 μm en promedio. Por otro lado analizaron los niveles de transcrito de los genes s6k (cinasa blanco de la protein-cinasa TOR) y cnaA (subunidad catalítica de la calceneurina), proteínas involucradas en procesos de diferenciación celular, entre otros.

Master thesis

CIENCIAS AGROPECUARIAS Y BIOTECNOLOGÍA IIQB-M-2017-1387 DNA RNA Proteína G

Ultraviolet-A Light Induces Micronucleated Erythrocytes in Newborn Rats

BLANCA PATRICIA LAZALDE RAMOS (2016)

Background: Ultraviolet-A (UV-A) light induce DNA damage by

creating pyrimidine dimers, or indirectly affects DNA by the formation

of reactive oxygen species. The objective was to determine DNA

damage by micronucleus test in neonatal rats exposed to UV-A

light.

Methods: Rat neonates were exposed to light from a LED

lamp (control group), to UV-C light 254 nm (control group to

desquamation skin) or UV-A light 365 nm and in one group the

dams were supplemented with folic acid (FA), to determine micro

nucleated erythrocytes (MNE) and micro nucleated polychromatic

erythrocytes (MNPCE) in peripheral blood of offspring.

Results: All the rat neonates exposed to UV-C lamp showed

desquamation skin, while for UV-A lamp no desquamation was

observed, and there was MNE differences in all sampling times

(P<0.02) and for MNPCE in 9 min group (P=0.001). No differences

between the groups with and without FA were observed.

Conclusion: Increased MNE frequencies without apparent damage

to the skin could be induced with UV-A light exposure. Under these

conditions, FA no protected against UV-A light exposure. This study

shows a manner to quantify the genotoxic effects of UV-A light in

peripheral blood erythrocytes of rat neonates.

Producción Científica de la Universidad Autónoma de Zacatecas UAZ

Article

BIOLOGÍA Y QUÍMICA UV-A light Folic acid Micronucleated erythrocytes DNA damage Neonates

Micronucleated erythrocytes in newborns rats exposed to three different types of ultraviolet-A (UVA) lamps from commonly uses devices

BLANCA PATRICIA LAZALDE RAMOS (2016)

Exposure to ultraviolet-A (UVA) light can accidentally cause adverse effects in the skin and eyes. UVA induces DNA damage directly by creating pyrimidine dimers or by the formation of reactive oxygen species that can indirectly affect DNA integrity. UVA radiation is emitted by lamps from everyday devices. In adult rats, micronucleated erythrocytes (MNE) are removed from the circulation by the spleen. However, in newborn rats, MNE have been observed in peripheral blood erythrocytes. The objective of this study was to use micronucleus tests to evaluate the DNA damage caused in newborn rats exposed to UVA light from three different types of UVA lamps obtained from commonly used devices: counterfeit detectors, insecticide devices, and equipment used to harden resins for artificial nails. Rat neonates were exposed to UVA lamps for 20 min daily for 6 days. The neonates were sampled every third day, and the numbers of MNE and micronucleated polychromatic erythrocytes (MNPCE) in the peripheral blood were determined. The rat neonates exposed to the three types of UVA lamps showed increased numbers of MNE and MNPCE from 48 h to 144 h (P < 0.05 and P < 0.001 respectively). However, no relationship was observed between the number of MNE and the wattage of the lamps. In conclusion, under these conditions, UVA light exposure induced an increase in MNE without causing any apparent damage to the skin.

Producción Científica de la Universidad Autónoma de Zacatecas UAZ

Article

BIOLOGÍA Y QUÍMICA Ultraviolet light Radiation DNA damage Micronuclei Erythrocytes

Iron oxide nanoparticles (IONPs) with potential applications in plasmid DNA isolation

José Raúl Sosa-Acosta SERGIO DIAZ CASTAÑON Julio César Zuaznabar Gardona (2018)

"DNA extraction and purification is considered a critical step in different biomedical applications such as genetic therapy and clinical diagnosis. This research describes the synthesis and characterization of functionalized IONPs with potential applications in plasmid DNA isolation. IONPs were synthesized by the chemical coprecipitation method followed by a post-synthesis functionalization using silica and (3-aminopropyl)triethoxysilane (APTES). A second functionalization strategy was carried out by an in situ coprecipitation of Fe(II) and Fe(III) ions in presence of chitosan and tris(hydroxymethyl)aminomethane (Tris). IONPs characterization by X-Ray diffraction (XRD) confirmed the synthesis of inverse-spinel magnetite like nanoparticles. In addition, infrared spectroscopy allowed to identify the hydroxyl, silanol and amino functional groups on the surface of the nanoparticles. Transmission electron microscopy measurements revealed IONPs with an average particle size under 13 nm. According to saturation and remanence magnetization values, all samples were suitable for bioseparation studies using magnetic manipulation. Preliminary separation assays with oligodeoxynucleotides (ODN) and plasmid DNA (pDNA) were carried out. Furthermore, biomolecular integrity of ODN and pDNA was verified using polyacrylamide and agarose gel electrophoresis, respectively. Synthesized IONPs and specially those functionalized with silica, chitosan and Tris presented comparable desorption percentages with some reported studies in plasmid DNA separation. Remarkably, by using such functionalized IONPs, the DNA desorption times were more than ten-time faster than other similar reported adsorbents. Therefore, they can be considered for DNA extraction and purification from complex biological samples."

Article

Plasmid DNA isolation Functionalized iron oxide nanoparticles Coprecipitation BIOLOGÍA Y QUÍMICA QUÍMICA QUÍMICA

TcG2/TcG4 DNA Vaccine Induces Th1 Immunity Against Acute Trypanosoma cruzi Infection: Adjuvant and Antigenic Effects of Heterologous T. rangeli Booster Immunization

JUAN CARLOS VAZQUEZ CHAGOYAN Nisha Garg Shivali Gupta BERENICE SALGADO JIMENEZ Nandadeva Lokugamage (2019)

Artículo Científico

Background: Chagas cardiomyopathy is caused by Trypanosoma cruzi (Tc). Two antigenic candidates, TcG2 and TcG4, are recognized by antibodies in naturally infected dogs and humans; and these vaccine candidates provided protection from Tc infection in mice and dogs. Trypanosoma rangeli (Tr) is non-pathogenic to mammals and shown to elicit cross-reactive anti-Tc antibodies. In this study, we investigated if fixed Tr (fTr) can further enhance the efficacy of the TcG2/TcG4 DNA vaccine. Methods and Results: C57BL/6 mice were immunized with TcG2/TcG4 DNA vaccine and fTr (delivered as an adjuvant or in prime-boost approach), and challenged with Tc. Serology studies showed that fTr (±quil-A) elicited Tc- and Tr-reactive IgGs that otherwise were not stimulated by TcG2/TcG4 vaccine only, and quil-A had suppressive effects on fTr-induced IgGs. After challenge infection, TcG2/TcG4-vaccinated mice exhibited potent expansion of antigen- and Tc-specific IgGs that were not boosted by fTr±quil-A. Flow cytometry analysis showed that TcG2/TcG4-induced dendritic cells (DC) and macrophages (M0) responded to challenge infection by expression of markers of antigen uptake, processing, and presentation, and production of pro-inflammatory cytokines. TcG2/TcG4-induced CD4+T cells acquired Th1 phenotype and expressed markers that orchestrate adaptive immunity. A fraction of vaccine-induced CD4+T cells exhibited iTreg phenotype responsible for aversion of self-injurious immune responses. Further, TcG2/TcG4-vaccinated mice exhibited potent expansion of poly-functional CD8+T cells with TNF-a/IFN-g production and cytolytic phenotype post-infection. Subsequently, tissue parasites and pathology were hardly detectable in TcG2/TcG4-vaccinated/infected mice. Inclusion of fTr±quil-A had no clear additive effects in improving the Tc-specific adaptive immunity and parasite control than was noted in mice vaccinated with TcG2/TcG4 alone. Non-vaccinated Gupta et al. Vaccine Testing Against T. cruzi mice lacked sufficient activation of Th1 CD4+/CD8+T cells, and exhibited >10-fold higher levels of tissue parasite burden than was noted in vaccinated/infected mice. Conclusion: TcG2/TcG4 vaccine elicits highly effective immunity, and inclusion of fTr is not required to improve the efficacy of DNA vaccine against acute Tc infection in mice.

UTMB

Article

Trypanosoma cruzi Chagas Disease Recombinant DNA Vaccine T. rangeli immune efficacy MEDICINA Y CIENCIAS DE LA SALUD