Understanding the spatial organization of the chromosomes in meiotic nuclei is crucial to our knowledge of the genome's functional regulation, stability and evolution. This study examined the nuclear architecture of Mus domesticus 2n=40 pachytene spermatocytes, analyzing the associations among autosomal bivalents via their Centromere Telomere Complexes (CTC). The study developed a nuclear model in which each CTC was represented as a 3D computer object. The probability of a given combination of associations among CTC was estimated by simulating a random distribution of 19 indistinguishable CTC over n indistinguishable "cells" on the nuclear envelope. The estimated association frequencies resulting from this numerical approach were similar to those obtained by quantifying actual associations in pachytene spermatocyte spreads. The nuclear localization and associations of CTC through the meiotic prophase in well-preserved nuclei were also analyzed. We concluded that throughout the meiotic prophase: 1) the CTC of autosomal bivalents are not randomly distributed in the nuclear space; 2) the CTC associate amongst themselves, probably at random, over a small surface of the nuclear envelope, at the beginning of the meiotic prophase; 3) the initial aggregation of centromere regions occurring in lepto-zygotene likely resolves into several smaller aggregates according to patterns of preferential partitioning; 4) these smaller aggregates spread over the inner face of the nuclear envelope, remaining stable until advanced stages of the meiotic prophase or even until the first meiotic division.
Considering the importance of the cutaneous tissue investigation and the need for the development of new protocols to non-invasively establish the safety and efficacy of dermocosmetics and topical products, we aimed at developing an HPLC-TBARS-EVSC (high performance liquid chromatography–thiobarbituric acid reactive species–ex vivo stratum corneum) assay for the lipid peroxidation measurement on subjects’ stratum corneum (SC) obtained by tape stripping; additionally, we applied the HPLC-TBARS-EVSC assay in an emulsified sunscreen system containing ethylhexyl triazone and bemotrizinol as UV filters. HPLC analysis was performed in isocratic mode with 35% methanol/65% phosphate buffer (pH 7.0) as the mobile phase. The diode detector was set at 532 nm to quantify the malondialdehyde (MDA)-TBA adduct. An ex vivo tape stripping method was applied in 10 volunteers in three pre-defined regions of the volar forearms: the control; the irradiated; and the site containing the sunscreen (2.0 mg·cm−2). Ten adhesive tapes per region were used for SC removal. An exclusive ex vivo protocol to measure SC lipid peroxidation was preliminarily developed with linearity and selectivity. The protocol suggested the use of an artificial irradiation dose (5506 KJ·m−2) to improve the assay response from the SC. The sunscreen system had a significative decrease in SC lipoperoxidative damage compared to the control. Our protocol can aid in the efficacy establishment of anti-UV and antioxidant agents, for instance, in studies that aim at elucidating the level of SC lipid peroxidation and even in carrying out baseline investigations characterizing different ethnicities and genders.
From tilapia (Oreochromis niloticus) farming, the by-products have been identified as a source of collagen that could be used for the development of dermocosmetics or pharmaceutical products. However, the characteristics of collagen related to a specific strain or culture must be well defined prior to its application. Collagen was extracted from the skin of three strains of tilapia: red YY males (YY: two Y-type sex chromosomes), XX gray females, and the F1: offspring of crossing red YY males with XX gray females; at different ages in the adult phase, using acetic acid and pepsin enzyme. The characteristics of acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) were shown by SDS-PAGE band profiles to be similar to bovine collagen type I (SIGMA), the PSC of gray tilapia being more fragile to temperature changes, consistent with the results of fractional viscosity. The characteristics of the F1 progeny were prioritized for being a commercially productive and sustainable source for the extraction of collagen, and the ASC form, being the one with the greatest stability and advantage over PSC, of importance to our investigations, leads to a controlled digestion as in the case of peptide induction, and also in the development of natural products in the pharmaceutical and/or dermocosmetic industry. Evaluations of the triple helix structure by FT-IR, X-ray diffraction and UV–visible spectroscopy give similar results between the strains: red, gray, and F1, and between ages in the adult form F1 (15, 24, and 36 months of age). Consequently, the skin of tilapia in adult form is recommended sustainably for up to 24 months of age where the collagen is obtained with the use of acetic acid without enzymatic treatment.
The influence of sex-related chromosomal arrangement in an YY and XY population of Oreochromis niloticus males was investigated in this study. A better spermatic mobility, motility, viability, confluence and less abnormality were found in double Y-chromosome males (YY-males), compared to XY-males; however, this better spermatic quality did not correlate with the body weight of individuals which was significantly better in XY-males. This study establishes the influence of YY-related sequences over spermatic quality and indicates the usefulness of the YY technology as a strategy to obtain better fish breeders and genetically male individuals for a tilapia farming program.
TRABAJOS ORIGINALES© Los autores. Este artículo es publicado por la Revista Peruana de Biología de la Facultad de Ciencias Biológicas, Universidad Nacional Mayor de San Marcos. Este es un artículo de acceso abierto, distribuido bajo los términos de la Licencia Creative Commons Atribución-NoComercial-CompartirIgual 4.0 Internacional.(http://creativecommons.org/licenses/by-nc-sa/4.0/), que permite el uso no comercial, distribución y reproducción en cualquier medio, siempre que la obra original sea debidamente citadas. Para uso comercial, por favor póngase en contacto con editor.revperubiol@gmail.com. Fuentes de financiamiento: El presente trabajo se realizó gracias al financiamiento del canon minero otorgado a la Universidad Nacional de Trujillo. Información sobre los autores:MA que realizó la parte experimental. LS-T realizó la parte experimental. ZP realizo el diseño experimental y planificación; MA, LS-T, ZP revisaron y aprobaron el manuscrito.Los autores no incurren en conflictos de intereses. Presentado:04/01/2017 Aceptado:22/09/2017 Publicado online: 28/10/2017 ResumenEl objetivo del trabajo fue realizar la estandarización de los protocolos moleculares, diferenciar los linajes de la tilapia roja y gris mediante microsatélites de ADN y evaluar los marcadores SCAR-5F-X/5R y SCAR-5F/5R-Y como indicadores del sexo genético de O. niloticus. El microsatélite UNH106 permitió diferenciar genéticamente los linajes de la tilapia roja de la gris. Los microsaté-lites UNH136, UNH115 y UNH995 presentaron loci monomórficos tanto en la tilapia roja como en la gris en el lote poblacional del Centro Experimental de Genética de la Universidad Nacional de Trujillo. Se describen los tamaños de fragmentos para los microsatélites y del gen de referencia β actin. También se confirmó la efectividad de los marcadores SCAR como informativos en la determinación del sexo genético evaluados en hembras XX y YY y machos XY y YY de O. niloticus roja y hembras XX y machos XY de O. niloticus gris.Palabras clave: Tilapia; Oreochromis; microsatélites; marcadores SCAR; determinación del sexo. AbstractThe aims of this work was to develop a standardized molecular protocols, to differentiate red and gray tilapia lineages using DNA microsatellites and to evaluate SCAR-5F-X/5R and SCAR-5F/5R-Y markers associated with the phenotypic sex of O. niloticus. The UNH106 microsatellite allowed differentiating genetically the lineages of red tilapia from gray. The markers UNH136, UNH115 and UNH995 presented monomorphic loci in both the red and gray tilapia in the population stock of the Centro Experimental de Genética of the Universidad Nacional de Trujillo. Fragment sizes for microsatellites and the reference gene β actin are described. The effectiveness of SCAR markers as informative in the determination of the genetic sex in XX and YY females and XY y YY males of red tilapia and XX females and XY males of gray tilapia was also confirmed.
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