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Castro, C.

Publications and source records attributed to Castro, C..

3 recordsLinked to original sources

Statistical optimization of culture conditions to improve cell density of Komagataeibacter medellinensis NBRC 3288: the first step towards to optimize bacterial cellulose production

The cell density yield of Komagataeibacter medellinensis NBRC 3288 was optimized applying response surface methodology (RSM) for the first time with this strain. Four factors were evaluated: glucose, initial pH, MgSO4 and KH2PO4, these were analyzed in a fractional factorial design to verify which were important to cell density yield. The best treatment assayed showed a yield of 1.09 OD600 and analysis of variance (ANOVA) selected glucose, inital pH and KH2PO4 as the significant factors. Next, a new experimental region with center point at glucose 0.5 %(wt./v), initial pH at 5.58 and KH2PO4 0.058 %(wt./v) was obtained by steepest ascent method and used to optimize biomass yield by Box-Behnken design. The results indicate that the optimum culture medium conditions predicted by the mathematical model (R2adj=72.09 %) were: glucose, 0.54 %(wt./v); peptone, 0.5 %(wt./v); yeast extract, 0.5 %(wt./v); KH2PO4, 0.059 %(wt./v); MgSO4, 0.025 %(wt./v); NaH2PO4, 0.267 %(wt./v) and pH, 5.18 (adjusted with citric acid, 0.2 %(wt./v)) to obtain an optimum cell density yield of 2.85 OD600 with an improve of 68 % respect to the best result obtained at the begin, as well as m improved 34.78 % and a reduction in cost of culture medium due to glucose concentration reduction of 75 %.\n\nImportanceDespite the important role of producer-strain play in bacterial cellulose production, until now there is not anu report about how to improve cell density, as a primary stage, that allows reach higher production of BC in later stages. In this study, was optimized the growth conditions of K. medellinensis (a BC producer in acid pH) in order to increase the cell density, as the primer stage in the whole process to optimize the production of BC, which is a very important supplie in different fields of science such as: food, materials, water treatment, tissue engineering, etc. The optimization process was achieved with savings in production costs of the culture medium, through the reduction of glucose concentration by 75 % and through the reduction of the microorganisms growth time.

microbiology

Separation and Loss of Centrioles from Primordidal Germ Cells to Mature Oocytes in the Mouse

Oocytes, including those from mammals, lack centrioles, but neither the mechanism by which mature eggs lose their centrioles nor the exact stage at which centrioles are destroyed during oogenesis is known. To answer questions raised by centriole disappearance during oogenesis, using a transgenic mouse expressing GFP-centrin-2 (GFP CETN2), we traced their presence from e11.5 primordial germ cells (PGCs) through oogenesis and their ultimate dissolution in mature oocytes. We show tightly coupled CETN2 doublets in PGCs, oogonia, and pre-pubertal oocytes. Beginning with follicular recruitment of incompetent germinal vesicle (GV) oocytes, through full oocyte maturation, the CETN2 doublets separate within the pericentriolar material (PCM); concomitantly, a rise in single CETN2 pairs is identified. CETN2 dissolution accelerates following meiosis resumption. Remarkably, a single CETN2 pair is retained in the PCM of most meiotic metaphase-I and -II spindle poles. Partial dissolution of the CETN2 foci occurs even as other centriole markers, like Cep135, a protein necessary for centriole duplication, are maintained at the PCM. Furthermore, live imaging demonstrates that the link between the two centrioles breaks as meiosis resumes and that centriole association with the PCM is progressively lost. Microtubule inhibition shows that centriole dissolution is uncoupled from microtubule dynamics. Thus, centriole doublets, present in early G2-arrested meiotic prophase oocytes, begin partial reduction during follicular recruitment and meiotic resumption, much later than previously thought.

developmental biology

Bead Enhancement of EV Analysis

Extracellular vesicles (EVs) are recognized cancer biomarkers, however, clinical analysis has been difficult due to a lack of simple and sensitive assays. Here, we describe a bead-enhanced flow cytometry method, BEAD flow, using biotinylated EVs captured on streptavidin particles. With this method, we show analysis of patient-derived EVs using a panel of pancreatic cancer biomarkers. BEAD flow is easily translatable to any biomarker or cancer type and can be run with conventional flow cytometers, making it highly flexible and adaptable to diverse research and clinical needs.

cancer biology