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1.
Biophys J ; 123(1): 31-41, 2024 01 02.
Artículo en Inglés | MEDLINE | ID: mdl-37968907

RESUMEN

DNA constructs for single-molecule experiments often require specific sequences and/or extrahelical/noncanonical structures to study DNA-processing mechanisms. The precise introduction of such structures requires extensive control of the sequence of the initial DNA substrate. A commonly used substrate in the synthesis of DNA constructs is plasmid DNA. Nevertheless, the controlled introduction of specific sequences and extrahelical/noncanonical structures into plasmids often requires several rounds of cloning on pre-existing plasmids whose sequence one cannot fully control. Here, we describe a simple and efficient way to synthesize 10.1-kb plasmids de novo using synthetic gBlocks that provides full control of the sequence. Using these plasmids, we developed a 1.5-day protocol to assemble 10.1-kb linear DNA constructs with end and internal modifications. As a proof of principle, we synthesize two different DNA constructs with biotinylated ends and one or two internal 3' single-stranded DNA flaps, characterize them using single-molecule force and fluorescence spectroscopy, and functionally validate them by showing that the eukaryotic replicative helicase Cdc45/Mcm2-7/GINS (CMG) binds the 3' single-stranded DNA flap and translocates in the expected direction. We anticipate that our approach can be used to synthesize custom-sequence DNA constructs for a variety of force and fluorescence single-molecule spectroscopy experiments to interrogate DNA replication, DNA repair, and transcription.


Asunto(s)
Proteínas de Ciclo Celular , ADN de Cadena Simple , Proteínas de Ciclo Celular/metabolismo , ADN/química , Replicación del ADN , Plásmidos/genética
2.
Aquac Nutr ; 2023: 5548991, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38192736

RESUMEN

Most commercial products available for Artemia sp. enrichment include high levels of docosahexaenoic acid (DHA) to boost its nutritional value, though with limited success. In this regard, the present study evaluated the alternative utilization of eicosapentaenoic acid (20 : 5n-3; EPA) oils to improve the n-3 long-chain highly unsaturated fatty acid content (n-3 LC-HUFA) in enriched Artemia sp. to feed greater amberjack (Seriola dumerili) larvae. Five experimental emulsions containing increasing levels of EPA from 0.8% to 60% of total fatty acids (TFA) and n-3 LC-HUFA (1.3%-70.6% TFA) were formulated. Each diet was fed to greater amberjack larvae (17-35 days posthatch (dph)) in three replicate 200-L tanks. The dietary EPA supplementation significantly improved larval growth during the feeding trial and survival at 35 dph (P < 0.05). In addition, larval fatty acid profiles showed a positive correlation with dietary EPA. Finally, despite the sum of total skeletal anomalies and column anomalies were not significantly affected by dietary EPA, increasing dietary EPA, and n-3 LC-HUFA tended to reduce the incidence of these types of anomalies in greater amberjack larvae at 35 dph. Based on these results, S. dumerili larvae could be successfully grown with low DHA but high EPA-rich oil enrichment products when n-3 LC-HUFA content in Artemia sp. is maintained in sufficient amounts.

3.
Aquac Nutr ; 2023: 5788432, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38130630

RESUMEN

Substitution of fish-based ingredients may alter the nutritional profile of the feeds, including the vitamin contents, ultimately leading to unbalanced vitamin supply. Vitamin A plays an essential role in epithelium preservation, cell differentiation, reproduction, and vision. It also intervenes in skeletogenesis through chondrocytes development. Therefore, low levels of vitamin A may cause poor growth and abnormal bone development among other symptoms. Besides, in gilthead seabream excess vitamin A altered bone structure and homeostasis, indicating that an upper level for vitamin A in feeds for this species must be defined. For this purpose, a practical plant-based diet (FM 10% and FO 6%) containing five increasing levels of vitamin A (24,000, 26,000, 27,000, 31,000, and 37,000 IU/kg) supplemented as retinyl acetate was formulated to identify the effects of high levels of vitamin A for gilthead seabream juveniles. The trial was conducted with 450 total fish distributed into 15 tanks, where each diet was tested in triplicates for 70 days. At the end of the trial, samples were taken for analyses of vitamin A-relevant markers. At the end of the trial the high levels of vitamin A supplementation did not cause a reduction in growth, whereas no significant effect was observed for the feed efficiency, specific growth rate, and feed convertion ratio. Although not significant, retinol content in liver showed a tendency to increase with the elevation of dietary vitamin A levels. Although minor, the highest level of vitamin A dietary content (37,000 IU/kg) caused a significant increase in caudal vertebrae partial fusion as well as caudal vertebrae malformations. Increasing dietary vitamin A was related to a reduction in the occurrence of microhemorrhages in the liver and a reduction in the presence of eosinophils associated to the pancreas. Overall, the results of the present study suggested that gilthead seabream juveniles fed a plant-based diet are able to tolerate very high levels of vitamin A supplementation when supplemented as retinyl acetate. Nevertheless, further supplementation should be avoided in order to reduce the prevalence of anomalies affecting the caudal vertebrae.

4.
EMBO J ; 37(5)2018 03 01.
Artículo en Inglés | MEDLINE | ID: mdl-29438978

RESUMEN

TDP-43 is an RNA-binding protein active in splicing that concentrates into membraneless ribonucleoprotein granules and forms aggregates in amyotrophic lateral sclerosis (ALS) and Alzheimer's disease. Although best known for its predominantly disordered C-terminal domain which mediates ALS inclusions, TDP-43 has a globular N-terminal domain (NTD). Here, we show that TDP-43 NTD assembles into head-to-tail linear chains and that phosphomimetic substitution at S48 disrupts TDP-43 polymeric assembly, discourages liquid-liquid phase separation (LLPS) in vitro, fluidizes liquid-liquid phase separated nuclear TDP-43 reporter constructs in cells, and disrupts RNA splicing activity. Finally, we present the solution NMR structure of a head-to-tail NTD dimer comprised of two engineered variants that allow saturation of the native polymerization interface while disrupting higher-order polymerization. These data provide structural detail for the established mechanistic role of the well-folded TDP-43 NTD in splicing and link this function to LLPS. In addition, the fusion-tag solubilized, recombinant form of TDP-43 full-length protein developed here will enable future phase separation and in vitro biochemical assays on TDP-43 function and interactions that have been hampered in the past by TDP-43 aggregation.


Asunto(s)
Proteínas de Unión al ADN/metabolismo , Agregación Patológica de Proteínas/genética , Dominios Proteicos/genética , Empalme del ARN/genética , Esclerosis Amiotrófica Lateral/genética , Esclerosis Amiotrófica Lateral/patología , Proteínas de Unión al ADN/genética , Células HEK293 , Humanos , Polimerizacion , Polímeros/metabolismo , Agregación Patológica de Proteínas/patología
5.
PLoS Comput Biol ; 17(7): e1009146, 2021 07.
Artículo en Inglés | MEDLINE | ID: mdl-34252083

RESUMEN

SARS-CoV-2 has spread across the world, causing high mortality and unprecedented restrictions on social and economic activity. Policymakers are assessing how best to navigate through the ongoing epidemic, with computational models being used to predict the spread of infection and assess the impact of public health measures. Here, we present OpenABM-Covid19: an agent-based simulation of the epidemic including detailed age-stratification and realistic social networks. By default the model is parameterised to UK demographics and calibrated to the UK epidemic, however, it can easily be re-parameterised for other countries. OpenABM-Covid19 can evaluate non-pharmaceutical interventions, including both manual and digital contact tracing, and vaccination programmes. It can simulate a population of 1 million people in seconds per day, allowing parameter sweeps and formal statistical model-based inference. The code is open-source and has been developed by teams both inside and outside academia, with an emphasis on formal testing, documentation, modularity and transparency. A key feature of OpenABM-Covid19 are its Python and R interfaces, which has allowed scientists and policymakers to simulate dynamic packages of interventions and help compare options to suppress the COVID-19 epidemic.


Asunto(s)
COVID-19/prevención & control , Trazado de Contacto , Análisis de Sistemas , COVID-19/epidemiología , COVID-19/transmisión , COVID-19/virología , Prueba de COVID-19 , Vacunas contra la COVID-19/administración & dosificación , Brotes de Enfermedades , Humanos , Distanciamiento Físico , Cuarentena , SARS-CoV-2/aislamiento & purificación
6.
Fish Shellfish Immunol ; 124: 244-253, 2022 May.
Artículo en Inglés | MEDLINE | ID: mdl-35421573

RESUMEN

Bacillus spp. supplementation as probiotics in cultured fish diets has a long history of safe and effective use. Specifically, B. velezensis show great promise in fine-tuning the European sea bass disease resistance against the pathogenicity caused by several members of the Vibrio family. However, the immunomodulatory mechanisms behind this response remain poorly understood. Here, to examine the inherent immune variations in sea bass, two equal groups were fed for 30 days with a steady diet, with one treatment supplemented with B. velezensis. The serum bactericidal capacity against live cells of Vibrio anguillarum strain 507 and the nitric oxide and lysozyme lytic activities were assayed. At the cellular level, the phagocytic response of peripheral blood leukocytes against inactivated Candida albicans was determined. Moreover, head-kidney (HK) total leukocytes were isolated from previously in vivo treated fish with LPS of V. anguillarum strain 507. Mechanistically, the expression of some essential proinflammatory genes (interleukin-1 (il1b), tumor necrosis factor-alpha (tnfa), and cyclooxygenase 2 (cox2) and the sea bass specific antimicrobial peptide (AMP) dicentracin (dic) expressions were assessed. Surprisingly, the probiotic supplementation significantly increased all humoral lytic and cellular activities assayed in the treated sea bass. In addition, time-dependent differences were observed between the control and probiotic treated groups for all the HK genes markers subjected to the sublethal LPS dose. Although the il1b was the fastest responding gene to a significant level at 48 h post-injection (hpi), all the other genes followed 72 h in the probiotic supplemented group. Finally, an in vivo bacteria challenge against live V. anguillarum was conducted. The probiotic fed fish observed a significantly higher survival. Overall, our results provide clear vertical evidence on the beneficial immune effects of B. velezensis and unveil some fundamental immune mechanisms behind its application as a probiotic agent in intensively cultured European sea bass.


Asunto(s)
Bacillus , Lubina , Enfermedades de los Peces , Vibriosis , Animales , Suplementos Dietéticos , Resistencia a la Enfermedad , Lipopolisacáridos , Vibrio , Vibriosis/veterinaria
7.
J Sports Sci ; 40(12): 1308-1314, 2022 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-35640042

RESUMEN

It is unknown whether ultrasound findings and symptoms of Achilles tendinopathy in runners correlate with foot strike patterns. We aimed to examine the relationships among Achilles tendon ultrasound findings in runners with or without Achilles tendinopathy, their foot strike patterns, and their training regimens. We recruited marathon runners 18 years of age or older with no history of Achilles tendon pain or surgery participating in the 2018 DONNA Marathon. Participants completed surveys and underwent Achilles tendon sonographic evaluations and were categorized by foot strike patterns. Seventy-nine runners were included; 22 (28%) with forefoot, 30 (38%) midfoot, and 27 (34%) hindfoot strike patterns. Foot strike pattern was not associated with tendon hyperaemia (P = 1.00) or hypoechogenicity (P = .97), and there was no association of cross-sectional area of the Achilles tendon with peak weekly distance while training. Sonographic characteristics of Achilles tendinopathy did not correlate with foot strike patterns or training regimens. Although not statistically significant, it is worth noting that cross-sectional area was 1 mm2 larger per every 1 kg/m2 increase in body mass index.


Asunto(s)
Tendón Calcáneo , Carrera , Tendinopatía , Tendón Calcáneo/diagnóstico por imagen , Adolescente , Adulto , Humanos , Carrera de Maratón , Autoinforme , Tendinopatía/diagnóstico por imagen
8.
Fish Shellfish Immunol ; 110: 100-115, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33444738

RESUMEN

Skin mucus is considered the first barrier against diseases in fish. The skin mucus protein profile of the greater amberjack (Seriola dumerili) and its changes due to experimental infection with Neobenedenia girellae were studied by combining 2-DE-MS/MS and gel-free LC-MS/MS proteomic approaches. The 2-DE results led to the identification of 69 and 55 proteins in noninfected and infected fish, respectively, and revealed that keratins were specifically cleaved in parasitized fish. Therefore, the skin mucus of the infected fish showed a higher protease activity due to, at least in part, an increase of metal-dependent protease and serine-type protease activities. Additionally, through a gel-free LC-MS/MS analysis, 1377 and 1251 different proteins were identified in the skin mucus of healthy and parasitized fish, respectively. The functional analysis of these proteins demonstrated a statistical overrepresentation of ribosomal proteins (a well-known source of antimicrobial peptides) in N. girellae-infected fish. In contrast, the components of membranes and protein transport GO categories were underrepresented after infection. Immune system process-related proteins constituted 2.5% of the total skin mucosal proteins. Among these skin mucosal proteins, 14 and 15 proteins exclusive to non-parasitized and parasitized fish were found, respectively, including specific serine-type proteases and metalloproteases in the parasitized fish. Moreover, the finding of tryptic peptides exclusive to some bacterial genera, obtained by gel-free LC-MS/MS, allowed us to construct a preliminary map of the microbiota living in the mucus of S. dumerili, with Pseudomonas and Paracoccus the most represented genera in both noninfected and infected fish.


Asunto(s)
Enfermedades de los Peces/inmunología , Proteínas de Peces/inmunología , Peces/inmunología , Péptido Hidrolasas/inmunología , Proteoma/inmunología , Piel/enzimología , Animales , Enfermedades de los Peces/parasitología , Microbiota , Moco/enzimología , Moco/metabolismo , Moco/microbiología , Piel/metabolismo , Piel/microbiología , Trematodos/fisiología , Infecciones por Trematodos/inmunología , Infecciones por Trematodos/parasitología , Infecciones por Trematodos/veterinaria
9.
Aquaculture ; 532: 736004, 2021 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-39175494

RESUMEN

The use of effective biocides as disinfectants is essential in aquaculture facilities. However, while most biocides act effectively on free-living planktonic pathogens, they are seldom useful against biofilms. In this study, we evaluate the biocidal efficacy and antimicrobial specific contact time of three disinfectants, Virkon™Aquatic (VirA), peracetic acid (PerA) and hydrogen peroxide (HydP), on Vibrio anguillarum, V. harveyi, V. alginolyticus, and Photobacterium damselae subspecies piscicida against their both life phases. By using the minimum inhibitory, bactericidal, and eradication concentrations of disinfectants acting on the free-living planktonic state (MIC; MBC) and biofilms (MBIC; MBEC), we determined the in vitro susceptibility of each bacterial strain against three different individual concentrations of VirA, PerA, and HydP added at 1, 5, and 10 min intervals. PerA and VirA had the highest bactericidal efficacies against the free-living planktonic state and biofilm of all bacteria. Kinetically, PerA gave a positive result more quickly in both cases regardless of the strain in question, while the weakest HydP required longer than 10 min to act effectively. Moreover, we conducted a short in vivo safety trial by pouring the suggested MIC of each disinfectant into tanks containing juvenile Gilthead seabream (Sparus aurata). A significant mortality after 24 h was observed pointing to the potential risk a mishap of these chemicals might cause to fish. Nevertheless, collectively, our results support the inclusion of biocides within biosecurity protocols in aquaculture facilities and highlight PerA as the most effective disinfectant for fighting against biofilms produced by V. anguillarum, V. harveyi, V. alginolyticus or P. damselae subsp. piscicida.

10.
Br J Nutr ; 122(1): 25-38, 2019 07 14.
Artículo en Inglés | MEDLINE | ID: mdl-31266551

RESUMEN

The present study investigated the effects of nutritional programming through parental feeding on offspring performance and expression of selected genes related to stress resistance in a marine teleost. Gilthead seabream broodstock were fed diets containing various fish oil (FO)/vegetable oil ratios to determine their effects on offspring performance along embryogenesis, larval development and juvenile on-growing periods. Increased substitution of dietary FO by linseed oil (LO) up to 80 % LO significantly reduced the total number of eggs produced by kg per female per spawn. Moreover, at 30 d after hatching, parental feeding with increasing LO up to 80 % led to up-regulation of the fatty acyl desaturase 2 gene (fads2) that was correlated with the increase in conversion rates of related PUFA. Besides, cyclo-oxygenase 2 (cox2) and TNF-α (tnf-α) gene expression was also up-regulated by the increase in LO in broodstock diets up to 60 or 80 %, respectively. When 4-month-old offspring were challenged with diets having different levels of FO, the lowest growth was found in juveniles from broodstock fed 100 % FO. An increase in LO levels in the broodstock diet up to 60LO raised LC-PUFA levels in the juveniles, regardless of the juvenile's diet. The results showed that it is possible to nutritionally programme gilthead seabream offspring through the modification of the fatty acid profiles of parental diets to improve the growth performance of juveniles fed low FO diets, inducing long-term changes in PUFA metabolism with up-regulation of fads2 expression. The present study provided the first pieces of evidence of the up-regulation of immune system-related genes in the offspring of parents fed increased FO replacement by LO.


Asunto(s)
Alimentación Animal/análisis , Dieta/veterinaria , Ácido Graso Desaturasas/metabolismo , Ácidos Grasos Insaturados/farmacología , Dorada/metabolismo , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Ácido Graso Desaturasas/genética , Ácidos Grasos Insaturados/administración & dosificación , Ácidos Grasos Insaturados/metabolismo , Femenino , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Metabolismo de los Lípidos/efectos de los fármacos , Fenómenos Fisiologicos Nutricionales Maternos
11.
Fish Shellfish Immunol ; 86: 35-45, 2019 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-30339845

RESUMEN

The main objective of this study was to determine the effect of two forms of mannan oligosaccharides (MOS: Bio-Mos® and cMOS: Actigen®, Alltech Inc, USA) and their combination on greater amberjack (Seriola dumerili) growth performance and feed efficiency, immune parameters and resistance against ectoparasite (Neobenedenia girellae) infection. Fish were fed for 90 days with 5 g kg-1 MOS, 2 g kg-1 cMOS or a combination of both prebiotics, in a Seriola commercial base diet (Skretting, Norway). At the end of the feeding period, no differences were found in growth performance or feed efficiency. Inclusion of MOS also had no effect on lysozyme activity in skin mucus and serum, but the supplementation of diets with cMOS induced a significant increase of serum bactericidal activity. Dietary cMOS also reduced significantly greater amberjack skin parasite levels, parasite total length and the number of parasites detected per unit of fish surface following a cohabitation challenge with N. girellae, whereas no effect of MOS was detected on these parameters. Of 17 immune genes studied cMOS dietary inclusion up-regulated hepcidin, defensin, Mx protein, interferon-γ (IFNγ), mucin-2 (MUC-2), interleukin-1ß (IL-1B), IL-10 and immunoglobulin-T (IgT) gene expression in gills and/or skin. MOS supplementation had a larger impact on spleen and head kidney gene expression, where piscidin, defensin, iNOS, Mx protein, interferons, IL-1ß, IL-10, IL-17 and IL-22 were all upregulated. In posterior gut dietary MOS and cMOS both induced IL-10, IgM and IgT, but with MOS also increasing piscidin, MUC-2, and IL-1ß whilst cMOS induced hepcidin, defensin and IFNγ. In general, the combination of MOS and cMOS resulted in fewer or lower increases in all tissues, possibly due to an overstimulation effect. The utilization of cMOS at the dose used here has clear benefits on parasite resistance in greater amberjack, linked to upregulation of a discrete set of immune genes in mucosal tissues.


Asunto(s)
Suplementos Dietéticos , Infestaciones Ectoparasitarias/veterinaria , Peces/inmunología , Regulación de la Expresión Génica/inmunología , Oligosacáridos/farmacología , Alimentación Animal , Animales , Dieta/veterinaria , Infestaciones Ectoparasitarias/inmunología , Infestaciones Ectoparasitarias/parasitología , Regulación de la Expresión Génica/efectos de los fármacos , Prebióticos , Distribución Aleatoria , Trematodos , Infecciones por Trematodos/inmunología , Infecciones por Trematodos/parasitología , Infecciones por Trematodos/veterinaria , Regulación hacia Arriba
14.
Br J Nutr ; 118(7): 500-512, 2017 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-28965514

RESUMEN

Nutrition during periconception and early development can modulate metabolic routes to prepare the offspring for adverse conditions through a process known as nutritional programming. In gilthead sea bream, replacement of fish oil (FO) with linseed oil (LO) in broodstock diets improves growth in the 4-month-old offspring challenged with low-FO and low-fishmeal (FM) diets for 1 month. The present study further investigated the effects of broodstock feeding on the same offspring when they were 16 months old and were challenged for a second time with the low-FM and low-FO diet for 2 months. The results showed that replacement of parental moderate-FO feeding with LO, combined with juvenile feeding at 4 months old with low-FM and low-FO diets, significantly (P<0·05) improved offspring growth and feed utilisation of low-FM/FO diets even when they were 16 months old: that is, when they were on the verge of their first reproductive season. Liver fatty acid composition was significantly affected by broodstock or reminder diets as well as by their interaction. Moreover, the reduction of long-chain PUFA and increase in α-linolenic acid and linoleic acid in broodstock diets lead to a significant down-regulation of hepatic lipoprotein lipase (P<0·001) and elongation of very long-chain fatty acids protein 6 (P<0·01). Besides, fatty acid desaturase 2 values were positively correlated to hepatic levels of 18 : 4n-3, 18 : 3n-6, 20 : 5n-3, 22 : 6n-3 and 22 : 5n-6. Thus, this study demonstrated the long-term nutritional programming of gilthead sea bream through broodstock feeding, the effect of feeding a 'reminder' diet during juvenile stages to improve utilisation of low-FM/FO diets and fish growth as well as the regulation of gene expression along the fish's life-cycle.


Asunto(s)
Alimentación Animal/análisis , Dieta/veterinaria , Metabolismo de los Lípidos , Dorada/crecimiento & desarrollo , Animales , Regulación hacia Abajo , Ácido Graso Desaturasas/genética , Ácido Graso Desaturasas/metabolismo , Ácidos Grasos/administración & dosificación , Ácidos Grasos Insaturados/administración & dosificación , Aceites de Pescado/administración & dosificación , Ácido Linoleico/administración & dosificación , Aceite de Linaza/administración & dosificación , Lipoproteína Lipasa/genética , Lipoproteína Lipasa/metabolismo , Hígado/metabolismo , Ácido alfa-Linolénico/administración & dosificación
16.
Gen Comp Endocrinol ; 252: 173-185, 2017 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-28652134

RESUMEN

The present study examined the short and mid-term effects of a rise in temperature from 18°C to 24°C on the expression of genes related to the stress response regulation in juveniles of Senegalese sole, Solea senegalensis. The animals were exposed to a temperature increase of 6°C, after 1month of acclimation at 18°C. After this process, samples of different tissues were collected from a total of 96 fish at four sampling points: 1h, 24h, 3days and 1week. The transcript levels of a set of genes involved in the stress response such as glucocorticoid receptors 1 and 2, corticotrophin-releasing factor, corticotrophin-releasing factor binding proteins, proopiomelanocortin A and B, and cellular stress defense (heat shock protein 70, 90AA and 90AB) were quantified at these sampling points. Additionally, blood samples were also taken to measure the circulating plasma cortisol concentration. Thermal stress induced by increasing temperature prompted an elevation of plasma cortisol levels in juvenile Senegalese sole after 1h as a short-term response, and a consecutive increase after one week, as a mid-term response. Senegalese sole seemed to respond positively in terms of adaptive mechanisms, with a rapid over-expression of grs and hsps in liver and brain, significantly higher after one hour post stress, denoting the fast and acute response of those tissues to a rapid change on temperature. The ratio hsp90/gr also increased 24h after thermal shock, ratio proposed to be an adaptive mechanism to prevent proteosomal degradation of GR. As a mid-term response, the elevation of brain crfbp gene expression one week after thermal shock could be an adaptive mechanism of negative feedback on HPI axis. Taken together, these data suggested an initial up-regulation of the glucocorticoid receptor complex linked genes in response to a temperature increase in Senegalese sole, with heat shock protein 90 potentially being a regulatory factor for the glucocorticoid receptor in the presence of cortisol.


Asunto(s)
Adaptación Fisiológica/genética , Peces Planos/genética , Peces Planos/fisiología , Regulación de la Expresión Génica , Receptores de Glucocorticoides/metabolismo , Estrés Fisiológico/genética , Temperatura , Animales , Peces Planos/sangre , Perfilación de la Expresión Génica , Respuesta al Choque Térmico/genética , Hidrocortisona/sangre , Especificidad de Órganos/genética
17.
J Prosthet Dent ; 117(2): 197-204, 2017 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-27666493

RESUMEN

This clinical report proposes a digital workflow using 2-dimensional (2D) digital photographs, a 3D extraoral facial scan, and cone beam computed tomography (CBCT) volumetric data to create a 3D virtual patient with craniofacial hard tissue, remaining dentition (including surrounding intraoral soft tissue), and the realistic appearance of facial soft tissue at an exaggerated smile under static conditions. The 3D virtual patient was used to assist the virtual diagnostic tooth arrangement process, providing patient with a pleasing preoperative virtual smile design that harmonized with facial features. The 3D virtual patient was also used to gain patient's pretreatment approval (as a communication tool), design a prosthetically driven surgical plan for computer-guided implant surgery, and fabricate the computer-aided design and computer-aided manufacturing (CAD-CAM) interim prostheses.


Asunto(s)
Diseño Asistido por Computadora , Implantación Dental Endoósea/métodos , Diseño de Prótesis Dental/métodos , Prótesis Dental de Soporte Implantado/métodos , Dentadura Parcial Provisoria , Cirugía Asistida por Computador/métodos , Tomografía Computarizada de Haz Cónico/métodos , Prótesis Dental , Femenino , Humanos , Procesamiento de Imagen Asistido por Computador , Imagenología Tridimensional , Persona de Mediana Edad
18.
Fish Shellfish Immunol ; 55: 662-70, 2016 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-27368538

RESUMEN

The generation of a variety of new therapeutic agents to control and reduce the effects of pathogen in aquaculture is urgently needed. The antimicrobial peptides (AMPs) are one of the major components of the innate defenses and typically have broad-spectrum antimicrobial activity. However, absorption and distributions of exogenous AMPs for therapeutics application on farmed fish species need to be studied. Previous studies in our laboratory have shown the properties of hepcidin as an effective antimicrobial peptide produced in fish in response to LPS and iron. Therefore, we decided to investigate the antimicrobial activity of four synthetic variants of hepcidin against Vibrio anguillarum in vitro, and using the more effective peptide we demonstrated the pathogen's ability to protect against the infection in European Sea bass. Additionally the uptake of this peptide after ip injection was demonstrated, reaching its distribution organs such as intestine, head kidney, spleen and liver. The synthetic peptide did not show cytotoxic effects and significantly reduced the accumulated mortalities percentage (23.5%) compared to the European Sea bass control (72.5%) at day 21. In conclusion, synthetic hepcidin shows antimicrobial activity against V. anguillarum and the in vivo experiments suggest that synthetic hepcidin was distributed trough the different organs in the fish. Thus, synthetic hepcidin antimicrobial peptide could have high potential for therapeutic application in farmed fish species.


Asunto(s)
Péptidos Catiónicos Antimicrobianos/farmacología , Lubina , Enfermedades de los Peces/prevención & control , Proteínas de Peces/farmacología , Hepcidinas/farmacología , Vibriosis/veterinaria , Vibrio/efectos de los fármacos , Animales , Enfermedades de los Peces/microbiología , Vibriosis/microbiología , Vibriosis/prevención & control
19.
An Acad Bras Cienc ; 88(3): 1191-200, 2016 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-27598839

RESUMEN

This study describes for the first time the chemical composition and olfactive description of floral scent from Brazilian Passiflora (Passiflora edulis Sim, Passiflora alata Curtis, Passiflora cincinnata Mast., Passiflora coccinea Aubl. and Passiflora quadrangularis L.). Five species were grown in greenhouse at the Agronomic Institute (IAC), São Paulo, Brazil. Volatile compounds were collected using dynamic headspace. Analyses of scent composition were performed by gas chromatograph coupled to mass spectrometer. Identification of chemical constituents was conducted through of retention index followed by comparative analysis of mass spectra with specialized databases. The olfactive descriptions of floral scent from each species was evaluated for a professional perfumer. High interspecific diversity was found between chemical compositions of floral scent within Passiflora and different bouquets were observed amount the studied species. Mayor constituents were linalool (P. alata), geraniol (P. quadrangularis), 1,4-dimethoxybenzene (P. edulis), benzaldehyde (P. cincinnata) and 2-methyl-3-pentanone (P. coccinea).


Asunto(s)
Flores/química , Odorantes/análisis , Passiflora/química , Brasil , Cromatografía de Gases y Espectrometría de Masas , Especificidad de la Especie
20.
Fish Shellfish Immunol ; 42(2): 508-16, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25447638

RESUMEN

The study assesses the effects of dietary concentrated mannan oligosaccharides (cMOS) on fish performance, biochemical composition, tissue fatty acid profiles, liver and posterior gut morphology and gen expression of selected parameters involved on the intestinal immune response and liver lipid metabolism of European sea bass (Dicentrarchus labrax). For that purpose, specimens of 20 g were fed during 8 weeks at 0 and 1.6 g kg(-1) dietary cMOS of inclusion in a commercial sea bass diet. Dietary cMOS enhanced fish length, specific and relative growth without affecting tissue proximate composition. However, cMOS supplementation altered especially liver and muscle fatty acid profiles by reducing levels of those fatty acids that are preferential substrates for ß-oxidation in spite of a preferential retention of long chain polyunsaturated fatty acids (LC-PUFA), such as 20:4n-6 or 22:5n-6, in relation to the down-regulation of delta 6/5 desaturase gene expression found in liver. Besides, dietary cMOS supplementation reduced posterior gut intestinal folds width and induced changes on the gene expression level of certain immune-related genes mainly by down regulating transforming growth factor ß (TGFß) and up-regulating immunoglobulin (Ig), major histocompatibility complex class II (MHCII), T cell receptor ß (TCRß) and Caspase 3 (Casp-3). Thus, dietary cMOS inclusion at 0.16% promoted European sea bass specific growth rate and length, stimulated selected cellular GALT-associated parameters and affected lipid metabolism in muscle and liver pointing to a higher LC-PUFA accumulation and promoted ß-oxidation.


Asunto(s)
Lubina/fisiología , Mananos/farmacología , Oligosacáridos/farmacología , Alimentación Animal/análisis , Animales , Lubina/genética , Lubina/crecimiento & desarrollo , Lubina/inmunología , Dieta/veterinaria , Suplementos Dietéticos/análisis , Regulación de la Expresión Génica/efectos de los fármacos , Inmunidad Innata/efectos de los fármacos , Distribución Aleatoria
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