RESUMEN
Products derived from medicinal plants with antimicrobial activity are considered a promising alternative in the treatment of fungal infections. In this perspective, this study proposed to evaluate the antifungal activity of the dichloromethane fraction of Annona crassiflora Mart. against C. albicans strains. Tests were carried out to determine Minimum Inhibitory Concentration (MIC), Minimum Fungicide Concentration (MFC), microbial growth kinetics, fungal cell wall and membrane mechanisms of action, antifungal biofilm activity, and cytotoxic effects on human erythrocytes. The extract presented MIC and MFC values that ranged from 256 µg/mL to 1,024 µg/mL, with fungicidal activity in the microbial growth kinetics assay. The mechanism of action did not occur through damage to the cell wall or via binding to ergosterol in the membrane, though the fraction presents activity against biofilm and is not cytotoxic in human erythrocytes. The dichloromethane fraction of Annona crassiflora Mart. presented antifungal activity and reduced biofilm growth, without toxicity against human erythrocytes; however, further studies are needed to define its mechanism of action.
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Annona , Antifúngicos , Biopelículas , Candida albicans , Cloruro de Metileno , Pruebas de Sensibilidad Microbiana , Extractos Vegetales , Annona/química , Antifúngicos/farmacología , Candida albicans/efectos de los fármacos , Humanos , Extractos Vegetales/farmacología , Biopelículas/efectos de los fármacos , Eritrocitos/efectos de los fármacosRESUMEN
OBJECTIVE: Herpes zoster virus can cause inflammatory neuropathy of the facial nerve. However, studies evaluating the prevalence of this agent in peripheral facial palsy are heterogeneous regarding sample group selection, laboratory analysis method and variables studied. In addition, there are a lack of epidemiological data in the Brazilian population on this serological phenomenon in peripheral facial palsy. This study estimated herpes zoster reactivation prevalence in serological samples through chemiluminescence immunoassay for quantitative determination of specific antibodies directed against the virus. METHODS: This cross-sectional study sought to determine the prevalence of viral reactivation by herpes zoster in subjects with idiopathic peripheral facial palsy through analysis of serological samples over a year. RESULTS: Forty-seven patients (32 females and 15 males) participated. Severe paralysis was more common in older patients (p = 0.017). Facial pain (p = 0.02) and vertigo (p = 0.001) were related to a worse evolution of facial palsy. The rate of serological reactivation of the virus was 12.76 per cent. CONCLUSION: The rate of serological reactivation of herpes virus in idiopathic peripheral facial palsy in our population is similar to foreign literature data, suggesting similar aetiological mechanisms in the genesis of this morbidity.
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Parálisis de Bell , Parálisis Facial , Herpes Zóster , Anciano , Anticuerpos Antivirales , Parálisis de Bell/epidemiología , Estudios Transversales , Parálisis Facial/epidemiología , Femenino , Herpes Zóster/complicaciones , Herpes Zóster/diagnóstico , Herpes Zóster/epidemiología , Herpesvirus Humano 3/fisiología , Humanos , Masculino , PrevalenciaRESUMEN
BACKGROUND: Noncommunicable diseases (NCDs) lead to drastic metabolic alterations with associated energy balance and body weight changes, two related physiological processes regulated by the brain. Polyphenol-based treatments for NCDs have emerged as a promising therapy, which seems to involve the energy balance modulation. However, it remains unclear what the most effective polyphenols-based treatment is to attenuate adverse effects in the energy balance of NCDs. OBJECTIVES: This systematic review aimed to evaluate the literature on the metabolic and neurological effects of polyphenols-based treatment in rodent models of NCDs. METHODS: Literature search was carried out in the following databases: CINAHL, Medline/PubMed, SCOPUS, and Web of Science. For title and abstract screening, original papers with polyphenols exposure in rodents were selected. For full-text screening, studies with models of NCDs that reported metabolic and neurological outcomes when treated with polyphenols were selected for inclusion in this review. RESULTS: 23 articles, using individual compound (11 articles) or polyphenols extracts (12 articles), were included in this review: 5 articles using tea polyphenols, 12 articles using grape-derived polyphenols, 3 articles using the polyphenol quercetin, and 3 articles using other polyphenol sources. Most results agree on the beneficial effect of polyphenols in attenuating alterations in energy balance and body weight. Such effects were associated with neuroprotective responses in different brain areas including hippocampus and hypothalamus. CONCLUSION: In conclusion, this review shows that the treatment with polyphenols, especially resveratrol or quercetin, attenuates the adverse effects of NCDs on energy balance and are associated with neuroprotective effects.
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Enfermedades no Transmisibles , Polifenoles , Animales , Peso Corporal , Polifenoles/farmacología , Quercetina , RoedoresRESUMEN
AIMS: To evaluate the ability of the haloarchaeon Haloferax volcanii to produce Ag and Au nanoparticles (NPs) and to characterize the obtained material in order to find relevant properties for future potential applications. METHODS AND RESULTS: Nanoparticles were produced by incubating H. volcanii cells with the corresponding metal salt. In the presence of precursor salts, cultures evidenced a colour change associated to the formation of metallic nanostructures with plasmonic bands located in the visible range of the spectrum. X-ray fluorescence analysis confirmed the presence of Ag and Au in the NPs which were spherical, with average sizes of 25 nmol l-1 (Ag) and 10 nmol l-1 (Au), as determined by electronic microscopy. Fourier transformed infrared spectroscopy indicated that both types of NPs showed a stable protein capping. Ag NPs evidenced antibacterial activity and Au NPs improved the specificity of polymerase chain reaction reactions. Au and Ag NPs were able to reduce 4-nitrophenol when incubated with NaBH4 . CONCLUSIONS: Haloferax volcanii is able to synthesize metallic NPs with interesting properties for technological applications. SIGNIFICANCE AND IMPACT OF THE STUDY: Our data demonstrate the ability of H. volcanii to synthesize metal NPs and constitutes a solid starting point to deepen the study and explore novel applications.
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Oro/metabolismo , Haloferax volcanii/metabolismo , Nanopartículas del Metal/microbiología , Plata/metabolismo , Antibacterianos/biosíntesis , Antibacterianos/química , Antibacterianos/farmacología , Borohidruros/metabolismo , Oro/química , Oro/farmacología , Nanopartículas del Metal/química , Nitrofenoles/metabolismo , Tamaño de la Partícula , Plata/química , Plata/farmacologíaRESUMEN
Current antifungal drugs present poor effectiveness and there is no available vaccine for fungal infections. Thus, novel strategies to treat or prevent invasive mycosis, such as cryptococcosis, are highly desirable. One strategy is the use of immunomodulators of polysaccharide nature isolated from mushrooms. The purpose of the present work was to evaluate the immunostimulatory activity of ß-(1,3)-glucan-containing exopolysaccharides (EPS) from the edible mushrooms Auricularia auricula in phagocytes and mice infected with Cryptococcus neoformans. EPS triggered macrophages and dendritic cell activation upon binding to Dectin-1, a pattern recognition receptor of the C-type lectin receptor family. Engagement of Dectin-1 culminated in pro-inflammatory cytokine production and cell maturation via its canonical Syk-dependent pathway signaling. Furthermore, upon EPS treatment, M2-like phenotype macrophages, known to support intracellular survival and replication of C. neoformans, repolarize to M1 macrophage pattern associated with enhanced production of the microbicidal molecule nitric oxide that results in efficient killing of C. neoformans. Treatment with EPS also upregulated transcript levels of genes encoding products associated with host protection against C. neoformans and Dectin-1 mediated signaling in macrophages. Finally, orally administrated ß-glucan-containing EPS from A. auricular enhanced the survival of mice infected with C. neoformans. In conclusion, the results demonstrate that EPS from A. auricula exert immunostimulatory activity in phagocytes and induce host protection against C. neoformans, suggesting that polysaccharides from this mushroom may be promising as an adjuvant for vaccines or antifungal therapy.
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Agaricales/química , Criptococosis/prevención & control , Polisacáridos Fúngicos/inmunología , Fagocitos/efectos de los fármacos , Fagocitos/inmunología , beta-Glucanos/inmunología , Animales , Criptococosis/inmunología , Cryptococcus neoformans/inmunología , Citocinas/inmunología , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Células Dendríticas/microbiología , Factores Inmunológicos/farmacología , Lectinas Tipo C/inmunología , Enfermedades Pulmonares Fúngicas , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Macrófagos/microbiología , Ratones , Ratones Endogámicos C57BL , Fagocitos/microbiología , Transducción de Señal , beta-Glucanos/farmacologíaRESUMEN
AIMS: To examine the antioxidant activity of Bacterioruberin (Bctr)-rich extracts isolated from a hyperpigmented, genetically modified Haloferax volcanii strain (HVLON3) and to investigate the effect on cold-sensitive ram sperm cells. METHODS AND RESULTS: The strain HVLON3 produces higher Bctr amounts than most haloarchaea (220 ± 13 mg g-1 DW). HVLON3-Bctr extract has higher antioxidant activity than ß-carotene (threefold) as evaluated using 2,2 diphenyl-1-picrylhydrazyl combined with Electron Paramagnetic Resonance analysis (EC50 4·5 × 10-5 mol l-1 vs 13·9 × 10-5 mol l-1 respectively). Different concentrations of HVLON3-Bctr extracts were assayed on ram sperm after freezing/thawing and physiologically relevant parameters were examined. Extracts containing 7 and 20 µmol l-1 Bctr significantly improved cell viability (P < 0·0001), total and progressive motility (P < 0·0001) and sperm velocities (P = 0·0172 for curvilinear velocity VCL, P = 0·0268 for average path velocity VAP and P = 0·0181 for straight line velocity VSL) and did not affect other parameters evaluated. CONCLUSIONS: HVLON3 is an excellent source of natural microbial C50 carotenoids with applicability in Biotechnology, Biomedical and Veterinary fields. HVLON3 Bctr extract improves the quality of cryopreserved ram sperm cells and could be applied to increase insemination yields. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides an insight on the bioactive properties of a bioproduct derived from haloarchaea (carotenoids) which are so far underexploited.
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Antioxidantes/farmacología , Carotenoides/farmacología , Haloferax volcanii/química , Espermatozoides/efectos de los fármacos , Animales , Antioxidantes/metabolismo , Carotenoides/metabolismo , Criopreservación , Haloferax volcanii/genética , Haloferax volcanii/metabolismo , Masculino , Ovinos , Motilidad Espermática/efectos de los fármacos , Espermatozoides/citologíaRESUMEN
Modern genotyping techniques, such as SNP analysis and genotyping by sequencing (GBS), are hampered by poor DNA quality and purity, particularly in challenging plant species, rich in secondary metabolites. We therefore investigated the utility of a pre-wash step using a buffered sorbitol solution, prior to DNA extraction using a high salt CTAB extraction protocol, in a high throughput or miniprep setting. This pre-wash appears to remove interfering metabolites, such as polyphenols and polysaccharides, from tissue macerates. We also investigated the adaptability of the sorbitol pre-wash for RNA extraction using a lithium chloride-based protocol. The method was successfully applied to a variety of tissues, including leaf, cambium and fruit of diverse plant species including annual crops, forest and fruit trees, herbarium leaf material and lyophilized fungal mycelium. We consistently obtained good yields of high purity DNA or RNA in all species tested. The protocol has been validated for thousands of DNA samples by generating high data quality in dense SNP arrays. DNA extracted from Eucalyptus spp. leaf and cambium as well as mycelium from Trichoderma spp. was readily digested with restriction enzymes and performed consistently in AFLP assays. Scaled-up DNA extractions were also suitable for long read sequencing. Successful RNA quality control and good RNA-Seq data for Eucalyptus and cashew confirms the effectiveness of the sorbitol buffer pre-wash for high quality RNA extraction.
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ADN/normas , Eucalyptus/genética , Polimorfismo de Nucleótido Simple , ARN/normas , Trichoderma/genética , Tampones (Química) , Cámbium/genética , ADN/aislamiento & purificación , ADN de Hongos/aislamiento & purificación , ADN de Hongos/normas , ADN de Plantas/aislamiento & purificación , ADN de Plantas/normas , Técnicas de Genotipaje , Micelio/genética , Hojas de la Planta/genética , ARN/aislamiento & purificación , ARN de Hongos/normas , ARN de Planta/aislamiento & purificación , ARN de Planta/normas , Análisis de Secuencia de ADN , Análisis de Secuencia de ARN , Sorbitol/químicaRESUMEN
INTRODUCTION: Severe genital abnormalities such as urogenital sinus and urethral duplication with ectopic urethra in the rectum represent a major challenge in reconstructive urology. OBJECTIVE: We aimed to review our cases presenting with functional ectopic urethra implanted in the rectum that were treated through an ASTRA approach. METHODS: We reviewed the medical records of all patients who had undergone an ASTRA approach from 2005-2016. We collected data with interest to primary diagnosis, clinical presentation, additional procedure, immediate clinical outcome, complications, bowel habits after surgery, voiding and bladder emptying pattern and long term follow-up. RESULTS: Since 2005, we treated 6 cases using this method, consisting of 3 patients with congenital aphallia and 3 with Y-type urethral duplication. Two aphallia patients underwent De Castro's neophalloplasty with simultaneous anastomosis of proximal urethra to a tubed buccal mucosa neourethra and one had a neophalloplasty with transverse skin flaps and primary perineal urethrostomy. Two patients with Y-type urethral duplication had a complete urethroplasty performed (one end-to-end anastomosis and a two-stage repair). Last patient had a definitive perineal urethrostomy. At mean follow-up of 83.5 meses, only one patient voids through the urethra, four have a Mitrofanoff channel and two have a perineal urethrostomy. Immediate follow-up was uneventful and none of our patients had any bowel complications nor fecal incontinence. DISCUSSION: Domini et al. were the first proponents of the anterior sagittal transanorectal approach (ASTRA) as an alternative to classic Peña approach. Later, De Castro popularized specifically this technique as a relevant part of his neophalloplasty procedure to treat congenital aphallia. We did not find in this series any complications related to ASTRA technique in regards to bowel habits, fecal incontinence or infection but most of cases we tried to create a new anterior urethra and connect to the proximal stump failed. Moreover, we are skeptic to support simultaneous urethral repair when there is a dysplastic bulbar segment between the proximal stump and the end of the penile urethra normally at penoscrotal junction area by urethral duplication. Treatment at two-stages is also prone to fail in the midterm. At end, most of them will have either a Mitrofanoff or perineal urethral stoma as shown in our data. CONCLUSION: The ASTRA approach proved to be useful for cases of rectal implantation of the urethra. No complications related to anterior sphincter incision were documented. We believe it should be regarded as the treatment of choice for cases alike ours.
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Recto/anomalías , Recto/cirugía , Uretra/anomalías , Uretra/cirugía , Preescolar , Humanos , Lactante , Masculino , Estudios Retrospectivos , Procedimientos Quirúrgicos Urológicos/métodosRESUMEN
Children who suffer from cerebral palsy (CP) often present comorbidities in the form of oro-facial dysfunctions. Studies in animals have contributed to elaborate potential therapies aimed at minimising the chronic disability of the syndrome. To systematically review the scientific literature regarding the possible effects that experimental models of CP can have on oro-facial functions. Two independent authors conducted a systematic review in the electronic databases Medline, Scopus, CINAHL, Web of Science and Lilacs, using Mesh and Decs terms in animal models. The motor and sensory parameters of sucking, chewing and swallowing were considered as primary outcomes; reactivity odour, controlled salivation, postural control, head mobility during feeding and the animal's ability to acquire food were secondary outcomes. Ten studies were included in the present review. Most studies used rabbits as experimental models of CP, which was induced by either hypoxia-ischemia, inflammation or intraventricular haemorrhage. Oro-facial functions were altered in all experimental models of CP. However, we found more modifications in hypoxia-ischemia models overall. On the other hand, the model of inflammation was more effective to reproduce higher damage for coordinating sucking and swallowing. All of the CP experimental models that were assessed modified the oral functions in different animal species. However, further studies should be conducted in order to clarify the mechanisms underlying oro-facial damage in order to optimise treatment strategies for children who suffer from CP.
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Parálisis Cerebral/fisiopatología , Trastornos de Deglución/fisiopatología , Modelos Animales de Enfermedad , Animales , Parálisis Cerebral/patología , Comorbilidad , Deglución , Trastornos de Deglución/patología , Ingestión de Alimentos , Humanos , Espasticidad Muscular , SalivaciónRESUMEN
This study investigated the antinociceptive effects of (S)-(-)-perillyl alcohol (PA) on orofacial nociception in Swiss male mice using formalin-, capsaicin-, and glutamate-induced pain tests. For each test, eight animals per group were pre-treated intraperitoneally by a blinded investigator with PA (50 or 75mg/kg), morphine, or vehicle (saline+0.2% Tween 80). The treatment was performed before the induction of orofacial nociception by injecting formalin, capsaicin, or glutamate solution into the right area of the upper lip. The orofacial nociceptive behaviour was timed in all tests by an investigator who was blinded to the treatments. The statistical analysis was performed using confidence intervals (CI), the effect size, and power. PA blocked the orofacial nociceptive behaviour at both doses tested (P<0.05) similarly to morphine (P>0.05), in all tests. The effect size was high in the phase 1 formalin test for 50mg/kg PA (95% CI 0.48-2.31, power 84.6%) and 75mg/kg PA (95% CI 0.82-2.76, power 96.2%), in phase 2 for 75mg/kg PA (95% CI 0.44-2.26, power 82.3%), and in the glutamate test for 75mg/kg PA (95% CI 1.11-3.16, power 99.2%). These findings show strong evidence for the antinociceptive properties of PA in the orofacial region.
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Dolor Facial/tratamiento farmacológico , Monoterpenos/farmacología , Animales , Capsaicina , Formaldehído , Ácido Glutámico , Masculino , Ratones , Morfina/farmacología , Dimensión del DolorRESUMEN
Jatropha curcas is an oilseed crop renowned for its tolerance to a diverse range of environmental stresses. In Brazil, this species is grown in semiarid regions where crop establishment requires a better understanding of the mechanisms underlying appropriate seed, seedling and plant behaviour under water restriction conditions. In this context, the objective of this study was to investigate the physiological and cytological profiles of J. curcas seeds in response to imbibition in water (control) and in polyethylene glycol solution (osmoticum). Seed germinability and reactivation of cell cycle events were assessed by means of different germination parameters and immunohistochemical detection of tubulin and microtubules, i.e. tubulin accumulation and microtubular cytoskeleton configurations in water imbibed seeds (control) and in seeds imbibed in the osmoticum. Immunohistochemical analysis revealed increasing accumulation of tubulin and appearance of microtubular cytoskeleton in seed embryo radicles imbibed in water from 48 h onwards. Mitotic microtubules were only visible in seeds imbibed in water, after radicle protrusion, as an indication of cell cycle reactivation and cell proliferation, with subsequent root development. Imbibition in osmoticum prevented accumulation of microtubules, i.e. activation of cell cycle, therefore germination could not be resumed. Osmoconditioned seeds were able to survive re-drying and could resume germination after re-imbibition in water, however, with lower germination performance, possibly due to acquisition of secondary dormancy. This study provides important insights into understanding of the physiological aspects of J. curcas seed germination in response to water restriction conditions.
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Citoesqueleto/metabolismo , Germinación , Jatropha/fisiología , Presión Osmótica , Ciclo Celular , Jatropha/citología , Microtúbulos/metabolismo , Plantones/citología , Plantones/fisiología , Semillas/citología , Semillas/fisiología , Estrés Fisiológico , Tubulina (Proteína)/metabolismo , Agua/fisiologíaRESUMEN
The serotonergic system plays a crucial role in the energy balance regulation. Energy balance is mediated by food intake and caloric expenditure. Thus, the present study investigated the mechanisms that might be associated with fluoxetine treatment-induced weight reduction. Wistar male rat pups received daily injections with subcutaneous fluoxetine (Fx-group) or vehicle solution (Ct-group) from day 1 until 21 days of age. Several analyses were conducted to verify the involvement of mitochondria in weight reduction. We found that body weight in the Fx-group was lower compared to control. In association to lower fat mass in the Fx-group (25%). Neither neonatal caloric intake nor food intake reveals significant differences. Evaluating caloric expenditure (locomotor activity and temperature after stimulus), we did not observe differences in locomotor activity. However, we observed that the Fx group had a higher capacity to maintain body temperature in a cold environment compared with the Ct-group. Since brown adipose tissue-(BAT) is specialized for heat production and the rate of heat production is related to mitochondrial function, we found that Fx-treatment increases respiration by 36%, although after addition of GDP respiration returned to Ct-levels. Examining ROS production we observe that Fx-group produced less ROS than control group. Evaluating uncoupling protein (UCP) expression we found that Fx-treatment increases the expression by 23%. Taken together, our results suggest that modulation of serotonin system results in positive modulation of UCP and mitochondrial bioenergetics in brown fat tissue.
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Tejido Adiposo Pardo/metabolismo , Tejido Adiposo Blanco/metabolismo , Metabolismo Energético/efectos de los fármacos , Fluoxetina/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Canales Iónicos/biosíntesis , Proteínas Mitocondriales/biosíntesis , Animales , Masculino , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Proteína Desacopladora 1RESUMEN
This study investigated the effects of maternal separation on the feeding behavior of rats. A maternal separation model was used on postnatal day 1 (PND1), forming the following groups: in the maternal separation (MS) group, pups were separated from their mothers each day from PND1 to PND14, whereas in the control (C) group pups were kept with their mothers. Subgroups were formed to study the effects of light and darkness: control with dark and light exposure, female and male (CF and CM), and maternal separation with dark and light exposure, female and male (SDF, SDM, SLF and SLM). Female rats had higher caloric intake relative to body weight compared with male controls in the dark period only (CF=23.3±0.5 v. CM=18.2±0.7, P<0.001). Macronutrient feeding preferences were observed, with male rats exhibiting higher caloric intake from a protein diet as compared with female rats (CF=4.1±0.7, n=8 v. CM=7.0±0.5, n=8, P<0.05) and satiety development was not interrupted. Female rats had a higher adrenal weight as compared with male rats independently of experimental groups and exhibited a higher concentration of serum triglycerides (n=8, P<0.001). The study indicates possible phenotypic adjustments in the structure of feeding behavior promoted by maternal separation, especially in the dark cycle. The dissociation between the mother's presence and milk intake probably induces adjustments in feeding behavior during adulthood.
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Preferencias Alimentarias/fisiología , Preferencias Alimentarias/psicología , Privación Materna , Saciedad/fisiología , Animales , Animales Recién Nacidos , Conducta Alimentaria/fisiología , Conducta Alimentaria/psicología , Femenino , Masculino , Embarazo , Ratas , Ratas WistarRESUMEN
Here we analyze the outcomes of unlimited access to a low-energy (LE) diet in dams and their offspring. At 3 weeks' gestation, pregnant Wistar rats were divided into two groups: (1) the control group received a normoenergetic diet; and (2) the experimental group received the LE diet. In dams, lactation outcomes, food intake, body weight, plasma IGF-1, prealbumin, transferrin and retinol-binding protein levels were evaluated; in offspring, biometric and biochemical parameters and food intake were evaluated. No differences were observed during pregnancy. However, after lactation, dams that received the LE diet demonstrated significant reductions in body weight (P<0.05), plasma IGF-1 (P=0.01), prealbumin and visceral fat (P<0.001). Pups born to dams that received the LE diet demonstrated reduced body length and weight at weaning (P<0.001) and were lighter than the control animals at the end of the experimental period. Pups also demonstrated reduced plasma, low-density lipoprotein (P=0.04), triglycerides (P=0.002) and glucose levels (P<0.05), and differences were noted in visceral fat. These results indicate that feeding dams with LE diet during the reproductive period induces acute malnutrition and impairs the growth and development of offspring, as well as certain metabolic parameters.
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Fenómenos Fisiologicos de la Nutrición Prenatal , Animales , Peso Corporal , Ingestión de Alimentos , Metabolismo Energético , Femenino , Desarrollo Fetal , Desnutrición/etiología , Estado Nutricional , Valor Nutritivo , Embarazo , Ratas WistarRESUMEN
The effects of linseed oil (LO) and macadamia oil (MO) on the metabolic changes induced by a high-fat diet (HFD) rich in saturated fatty acid were investigated. For the purpose of this study, the vegetable oil present in the HFD, i.e. soybean oil (SO) was replaced with LO (HFD-LO) or MO (HFD-MO). For comparative purposes, a group was included, which received a normal fat diet (NFD). Male Swiss mice (6-week old) were used. After 14 days under the dietary conditions, the mice were fasted for 18 h, and experiments were then performed. The HFD-SO, HFD-LO and HFD-MO groups showed higher glycaemia (p < 0.05 versus NFD). However, no significant effect was observed on glycaemia, liver gluconeogenesis and liver ketogenesis when SO was replaced by either LO or MO. The body weight and the sum of epididymal, mesenteric, retroperitoneal and inguinal fat weights were higher (p < 0.05) in the HFD-SO and HFD-MO groups as compared with the NFD group. However, there was no significant difference in these parameters between the NFD and HFD-LO groups. Thus, the protective role of LO on lipid accumulation induced by an HFD rich in saturated fatty acid is potentially mediated by the high content of É·-3 polyunsaturated fatty acid in LO.
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Glucemia/metabolismo , Dieta Alta en Grasa , Lino , Hígado/metabolismo , Macadamia , Aceites de Plantas/farmacología , Animales , Peso Corporal , Ingestión de Energía , Gluconeogénesis , Cetosis/etiología , Metabolismo de los Lípidos , Masculino , Ratones , Aumento de PesoRESUMEN
AIMS: Differences in fibre-type composition of skeletal muscle have been associated with obesity and insulin resistance. As a poor nutrient environment early in life is a predisposing factor for the development of obesity and related metabolic diseases at adulthood, this study aimed at determining the long-term consequences of maternal undernutrition on the structural and metabolic properties of two skeletal muscles characterized by their different fibre-type composition and metabolic properties. METHODS: The fibre-type composition and enzymatic activities of hexokinase (HK), beta-hydroxyacyl-CoA dehydrogenase (ß-HAD) and citrate synthase (CS) were measured in soleus and extensor digitorum longus (EDL) muscles from adult rats born to dams fed a control (17% protein) or a low-protein [8% protein (PR)] diet throughout pregnancy and lactation. In addition, the expression levels of several genes regulating glycolysis, fatty acid oxidation and mitochondrial biogenesis were determined by real-time PCR. RESULTS: Protein rats exhibited enhanced density of type II fibres along with decreased rate of fatty acid oxidation and glycolysis in soleus but not EDL. Malnourished rats exhibited also a different gene expression profile in soleus and EDL. Altogether, these alterations correspond to a state of energy deficiency and are present in animals which do not show yet any sign of obesity or glucose intolerance. CONCLUSION: We conclude that maternal protein restriction alters in the long term the structural and enzymatic properties of offspring skeletal muscle in a fibre-type-dependent manner. These alterations might have a causative role in the development of obesity and related metabolic disorders later in life.
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Envejecimiento/metabolismo , Dieta con Restricción de Proteínas , Fibras Musculares de Contracción Rápida/enzimología , Fibras Musculares de Contracción Rápida/patología , Fibras Musculares de Contracción Lenta/enzimología , Fibras Musculares de Contracción Lenta/patología , Proteínas Musculares/metabolismo , 3-Hidroxiacil-CoA Deshidrogenasas/metabolismo , Envejecimiento/patología , Animales , Citrato (si)-Sintasa/metabolismo , Proteínas en la Dieta/metabolismo , Femenino , Hexoquinasa/metabolismo , Masculino , Ratas , Ratas WistarRESUMEN
Immobilization of small proteins designed to perform protein-protein assays can be a difficult task. Often, the modification of reactive residues necessary for the interaction between the immobilized protein and the matrix compromises the interaction between the protein and its target. In these cases, glutathione-S-transferase (GST) is a valuable tag providing a long arm that makes the bait protein accessible to the mobile flow phase of the chromatography. In the present report, we used a GST fusion version of the 8-kDa protein serine protease inhibitor Kazal-type 3 (SPINK3) as the bait to purify anti-SPINK3 antibodies from a rabbit crude serum. The protocol for immobilization of GST-SPINK3 to glutathione-agarose beads was modified from previously reported protocols by using an alternative bifunctional cross-linker (dithiobis(succinimidyl propionate)) in a very simple procedure and by using simple buffers under physiological conditions. We concluded that the immobilized protein remained bound to the column after elution with low pH, allowing the reuse of the column for alternative uses, such as screening for other protein-protein interactions using SPINK3 as the bait.
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Métodos Analíticos de la Preparación de la Muestra , Anticuerpos/aislamiento & purificación , Glutatión Transferasa/química , Proteínas Inmovilizadas/química , Proteínas/metabolismo , Proteínas Recombinantes de Fusión/química , Sefarosa/química , Anticuerpos/química , Glutatión/química , Glutatión Transferasa/metabolismo , Unión Proteica , Proteínas/química , Proteínas Recombinantes de Fusión/metabolismoRESUMEN
BACKGROUND: The contribution of liver glycogen catabolism to hyperglycemia and glucose intolerance induced by pharmacological hypercortisolism were investigated. METHODS: For this purpose, adult male Wistar rats that received 1.0 mg/kg dexamethasone (DEX) ip at 8:00 a.m. (DEX group) or saline (CON group) once a day for 5 consecutive days were compared. RESULTS: Experimental hypercortisolism was confirmed by higher (p<0.05) glycemia, lower (p<0.05) body weight and glucose intolerance. In the fed state, the basal glycogen catabolism and the glucagon (1 nM) and epinephrine (2 µM) induced glycogen catabolism were similar between the groups. The activation of glycogen catabolism induced by phenylephrine (2 µM) and isoproterenol (20 µM) were increased (p<0.05) and decreased (p<0.05), respectively, in DEX rats. Furthermore, DEX rats exhibited higher (p<0.05) glycogen catabolism during the infusion of cAMP (3 µM). However, during the infusion of cAMP (15 µM), 6MB-cAMP (3 µM) or cyanide (0.5 mM), the intensification of glycogen breakdown was similar. Thus, in general, hypercortisolism does not influence the basal glycogen catabolism and the liver responsiveness to glycogenolytic agents in the fed state. In contrast with fed state, fasted rats (DEX group) showed a more intense (p<0.05) basal glycogen catabolism. CONCLUSION: The contribution of glycogen catabolism to hyperglycemia during hypercortisolism depends of the nutritional status, starting from a negligible participation in the fed state up to a significant contribution in the fasted state.
Asunto(s)
Síndrome de Cushing/inducido químicamente , Dexametasona/toxicidad , Glucógeno/metabolismo , Hígado/efectos de los fármacos , Animales , Peso Corporal , Síndrome de Cushing/fisiopatología , AMP Cíclico/administración & dosificación , Dexametasona/administración & dosificación , Epinefrina/administración & dosificación , Ayuno , Glucagón/administración & dosificación , Intolerancia a la Glucosa , Hiperglucemia/inducido químicamente , Hiperglucemia/fisiopatología , Hígado/metabolismo , Masculino , Ratas , Ratas WistarRESUMEN
The aim of the study was to verify if moderate physical training affects leptin content in visceral and subcutaneous adipose tissue of adult rats subjected to a low-protein diet during the perinatal period. Male Wistar rats were divided into 2 groups according to their mother's diet during gestation and lactation: control (17% casein, C, n=12) and low-protein (8% casein, LP, n=12). On postnatal day 60, half of each group was submitted to moderate physical training (8 wks, 5 d · wk - 1, 60 min · d - 1, at 70% of VO2max, T) or not. After the physical training period, visceral and subcutaneous adipose tissues were removed. Leptin content was evaluated by western blotting. Starting from the fifth week on, T pups showed a reduction in the body weight. Similarly, LP+T offspring showed a lower body weight starting from the sixth week on. Western blotting analysis showed that leptin content in the visceral tissue was higher in the LP rats (p<0.01) and it was reversed in LP+T. No difference was found in the subcutaneous tissue. Moderate physical training attenuated the effects of a perinatal low-protein diet on the leptin content in visceral adipose tissue in adult offspring.
Asunto(s)
Ejercicio Físico , Grasa Intraabdominal/metabolismo , Leptina/biosíntesis , Animales , Dieta con Restricción de Proteínas , Femenino , Desarrollo Fetal , Humanos , Lactancia/metabolismo , Masculino , Fenómenos Fisiologicos Nutricionales Maternos , Atención Perinatal , Ratas , Ratas WistarRESUMEN
To determine the hemodynamic mechanisms responsible for the attenuated blood pressure response to mental stress after exercise, 26 healthy sedentary individuals (age 29 ± 8 years) underwent the Stroop color-word test before and 60 min after a bout of maximal dynamic exercise on a treadmill. A subgroup (N = 11) underwent a time-control experiment without exercise. Blood pressure was continuously and noninvasively recorded by infrared finger photoplethysmography. Stroke volume was derived from pressure signals, and cardiac output and peripheral vascular resistance were calculated. Perceived mental stress scores were comparable between mental stress tests both in the exercise (P = 0.96) and control (P = 0.24) experiments. After exercise, the blood pressure response to mental stress was attenuated (pre: 10 ± 13 vs post: 6 ± 7 mmHg; P < 0.01) along with lower values of systolic blood pressure (pre: 129 ± 3 vs post: 125 ± 3 mmHg; P < 0.05), stroke volume (pre: 89.4 ± 3.5 vs post: 76.8 ± 3.8 mL; P < 0.05), and cardiac output (pre: 7.00 ± 0.30 vs post: 6.51 ± 0.36 L/min; P < 0.05). Except for heart rate, the hemodynamic responses and the mean values during the two mental stress tests in the control experiment were similar (P > 0.05). In conclusion, a single bout of maximal dynamic exercise attenuates the blood pressure response to mental stress in healthy subjects, along with lower stroke volume and cardiac output, denoting an acute modulatory action of exercise on the central hemodynamic response to mental stress.