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1.
Arch Microbiol ; 206(1): 54, 2024 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-38180520

RESUMEN

Pseudomonas aeruginosa is an opportunistic bacterium that can form a biofilm with the ability to colonize different surfaces and for increasing resistance to antibiotics. An alternative to solve this problem may be the use of non-glucose/mannose glycosylated proteins from Melipona beecheii honey, which are capable of inhibiting the growth of this pathogen. In this work, the antibiofilm activity of the conA-unbound protein fraction (F1) from M. beecheii was evaluated. The crude protein extract (CPE) and the F1 fraction inhibited the P. aeruginosa biofilm growth above 80% at 4 and 1.3 µg/mL, respectively. These proteins affected the structure of the biofilm, as well as fleQ and fleR gene expressions involved in the formation and regulation of the P. aeruginosa biofilm. The results demonstrated that the F1 fraction proteins of M. beecheii honey inhibit and affect the formation of the P. aeruginosa biofilm.


Asunto(s)
Miel , Infecciones por Pseudomonas , Abejas , Animales , Pseudomonas aeruginosa/genética , Antibacterianos/farmacología , Biopelículas , Concanavalina A
2.
BMC Vet Res ; 20(1): 322, 2024 Jul 18.
Artículo en Inglés | MEDLINE | ID: mdl-39026205

RESUMEN

Total protein levels in fish are widely used in health and welfare studies, providing a simple and accessible measure. However, the multifaceted role of blood proteins makes it sometimes challenging to link total protein content to specific health issues, while specific protein fractions may offer more precise insights into fish biology and health, particularly in farmed fish species where such data is often lacking. Data were gathered from two experiments involving Dicentrarchus labrax and Sparus aurata, key species in European marine aquaculture. The aim was (1) to assess how different globulin fractions contribute to total protein content in blood and (2) how these contributions vary across different sampling times in healthy animals. In D. labrax, the beta1 globulin fraction emerged as the major contributor (34.16%), followed by albumin and alpha2 globulins (18.24% and 16.41%, respectively). In contrast, pre-albumins and alpha1 fractions had the least contribution (5.49% and 7.71%). S. aurata exhibited albumin as the primary contributor (23.39%), followed by beta1 and alpha2 globulins (19.71% and 19.15%, respectively), with gamma and alpha1 fractions contributing the least (5.34% and 8.63%). Notably, the study revealed relatively stable contributions of globulin fractions to total proteins within both species, albeit with minor variations over time, potentially linked to environmental and individual factors. Furthermore, larger fish displayed higher total protein levels. This research underscores the need for further investigation into the diverse factors influencing globulin contributions to total proteins, ultimately enhancing health and welfare monitoring for farmed fish species.


Asunto(s)
Lubina , Proteínas Sanguíneas , Dorada , Animales , Lubina/sangre , Dorada/sangre , Proteínas Sanguíneas/análisis , Acuicultura , Mar Mediterráneo
3.
Exp Parasitol ; 256: 108656, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38097154

RESUMEN

Infections with D. dendriticum are distributed worldwide and mostly associated with ruminant livestock. Depending on the length and strength of the infection it can be manifested with losses in milk production, reductions in milk and wool quality, decreased weight gains, reproductive performance and poor carcass quality. The objective of this study was to determine the efficacy of albendazole (ABZ) against the lancet liver fluke Dicrocoelium dendriticum in naturally infected sheep using parasitological methods. Twenty-four sheep were divided into four groups: two untreated control groups (C1, C2) and two treated groups (T1, T2), with six animals in each group. The sheep in the treated groups were administered a single oral dose (15 mg/kg bwt) of ABZ suspension. After ABZ treatment the animals were slaughtered on Day 14 (groups C1, T1) and Day 30 (groups C2, T2) and were necropsied. Coprological therapeutic ABZ efficacy reached 92.4% on Day 14 (P < 0.001) and 88.5% on Day 30 (P < 0.001). On Day 30, the serum activities of hepatic and cholestatic enzymes including serological analysis of total protein concentration (TP) and protein fractions were evaluated. Significant decrease of aspartate aminotransferase (AST) (P < 0.01) and gamma-glutamyltransferase (GGT) (P < 0.05) activity by 36.9% and 34.6%, respectively, were detected for sheep in T2 group. These enzymes showed a strong positive correlation to fluke burden: AST (r = 0.654) and GGT (r = 0.768), respectively (P < 0.05). Additionally, the electrophoretic analysis of serum total protein and protein fraction concentrations revealed minimal hypoproteinemia and hyperalbuminemia after ABZ treatment. The decrease of liver enzyme activities and their correlation with fluke burden may indicate recovery of hepatocellular and biliary damage following the reduction of fluke burdens after ABZ therapy. A decline in AST and GGT activity could serve as a valuable adjunct bioindicator of liver damage and fluke reduction after treatment of dicrocoeliosis in naturally infected sheep.


Asunto(s)
Dicrocoelium , Fasciola hepatica , Enfermedades de las Ovejas , Animales , Ovinos , Albendazol/farmacología , Albendazol/uso terapéutico , Dicrocoelium/metabolismo , Fasciola hepatica/metabolismo , Enfermedades de las Ovejas/tratamiento farmacológico , Hígado/metabolismo , gamma-Glutamiltransferasa
4.
Molecules ; 29(17)2024 Aug 26.
Artículo en Inglés | MEDLINE | ID: mdl-39274875

RESUMEN

The recovery of valuable nutritional compounds, like proteins, from waste streams and by-products is a key strategy for enhancing production sustainability and opening up new market potential. This research aimed to use high-intensity ultrasound as an innovative technique to extract the soluble proteins from the pumpkin leaves. The impact of various sonication amplitudes and duration periods on protein yield, functional properties, antioxidant qualities, and structural characteristics, were studied. Utilization of ultrasound technology significantly increased the yield of pumpkin leaf protein by up to 40%-six times higher than maceration. The ultrasound extraction provided a RuBisCO-rich protein fraction with high radical scavenging and chelating activities, especially at 40% amplitude. Cavitation modified the tertiary and secondary structures of leaf proteins: the amount of α-helix changed based on amplitude (12.3-37.7%), the amount of random coil increased to 20.4%, and the amount of ß-turn reduced from 31 to 18.6%. The alteration of the protein fluorescence spectrum (blue shift in spectrum) provides further evidence that ultrasound alters the proteins' molecular structure in comparation with maceration; the maximum tryptophan fluorescence intensity decreased from 22.000 to 17.096. The hydrophobicity values of 76.8-101.5 were substantially higher than the maceration value of 53.4, indicating that ultrasound improved the hydrophobicity of protein surfaces. Ultrasound resulted in a significant increase in solubility in an acidic environment with the increase in sonication amplitude. A 2.4-fold increase in solubility at pH 2 becomes apparent (20% amplitude; 43.1%) versus maceration (18.2%). The emulsifying ability decreases from 6.62 to 5.13 m2/g once the sonication amplitude increases by 20-70%. By combining the ultrasound periods and amplitudes, it is possible to create high-value protein leaf extracts with improved properties which can find real application as food additives and dietary supplements.


Asunto(s)
Cucurbita , Tecnología Química Verde , Hojas de la Planta , Proteínas de Plantas , Cucurbita/química , Hojas de la Planta/química , Proteínas de Plantas/química , Antioxidantes/química , Ondas Ultrasónicas , Sonicación/métodos , Interacciones Hidrofóbicas e Hidrofílicas
5.
J Sci Food Agric ; 2024 Jun 28.
Artículo en Inglés | MEDLINE | ID: mdl-38940359

RESUMEN

BACKGROUND: The increasing demand for sustainable alternatives to traditional protein sources, driven by population growth, underscores the importance of protein in a healthy diet. Pecan (Carya illinoinensis (Wangenh.) K. Koch) nuts are currently underutilized as plant-based proteins but hold great potential in the food industry. However, there is insufficient information available on pecan protein, particularly its protein fractions. This study aimed to explore the physicochemical and functional properties of protein isolate and the main protein fraction glutelin extracted from pecan nuts. RESULTS: The results revealed that glutelin (820.67 ± 69.42 g kg-1) had a higher crude protein content compared to the protein isolate (618.43 ± 27.35 g kg-1), while both proteins exhibited amino acid profiles sufficient for adult requirements. The isoelectric points of protein isolate and glutelin were determined to be pH 4.0 and pH 5.0, respectively. The denaturation temperature of the protein isolate (90.23 °C) was higher than that of glutelin (87.43 °C), indicating a more organized and stable conformation. This is further supported by the fact that the protein isolate had a more stable main secondary structure than glutelin. Both proteins demonstrated improved solubility, emulsifying, and foaming properties at pH levels deviating from their isoelectric points in U-shaped curves. Compared to the protein isolate, glutelin displayed superior water and oil absorption capacity along with enhanced gelling ability. CONCLUSION: The protein isolate and glutelin from pecan nuts exhibited improved stability and competitive functional properties, respectively. The appropriate utilization of these two proteins will support their potential as natural ingredients in various food systems. © 2024 Society of Chemical Industry.

6.
J Dairy Sci ; 106(3): 1853-1873, 2023 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-36710177

RESUMEN

In recent years, increasing attention has been focused on the genetic evaluation of protein fractions in cow milk with the aim of improving milk quality and technological characteristics. In this context, advances in high-throughput phenotyping by Fourier transform infrared (FTIR) spectroscopy offer the opportunity for large-scale, efficient measurement of novel traits that can be exploited in breeding programs as indicator traits. We took milk samples from 2,558 Holstein cows belonging to 38 herds in northern Italy, operating under different production systems. Fourier transform infrared spectra were collected on the same day as milk sampling and stored for subsequent analysis. Two sets of data (i.e., phenotypes and FTIR spectra) collected in 2 different years (2013 and 2019-2020) were compiled. The following traits were assessed using HPLC: true protein, major casein fractions [αS1-casein (CN), αS2-CN, ß-CN, κ-CN, and glycosylated-κ-CN], and major whey proteins (ß-lactoglobulin and α-lactalbumin), all of which were measured both in grams per liter (g/L) and proportion of total nitrogen (% N). The FTIR predictions were calculated using the gradient boosting machine technique and tested by 3 different cross-validation (CRV) methods. We used the following CRV scenarios: (1) random 10-fold, which randomly split the whole into 10-folds of equal size (9-folds for training and 1-fold for validation); (2) herd/date-out CRV, which assigned 80% of herd/date as the training set with independence of 20% of herd/date assigned as the validation set; (3) forward/backward CRV, which split the data set in training and validation set according with the year of milk sampling (FTIR and gold standard data assessed in 2013 or 2019-2020) using the "old" and "new" databases for training and validation, and vice-versa with independence among them; (4) the CRV for genetic parameters (CRV-gen), where animals without pedigree as assigned as a fixed training population and animals with pedigree information was split in 5-folds, in which 1-fold was assigned to the fixed training population, and 4-folds were assigned to the validation set (independent from the training set). The results (i.e., measures and predictions) of CRV-gen were used to infer the genetic parameters for gold standard laboratory measurements (i.e., proteins assessed with HPLC) and FTIR-based predictions considering the CRV-gen scenario from a bi-trait animal model using single-step genomic BLUP. We found that the prediction accuracies of the gradient boosting machine equations differed according to the way in which the proteins were expressed, achieving higher accuracy when expressed in g/L than when expressed as % N in all CRV scenarios. Concerning the reproducibility of the equations over the different years, the results showed no relevant differences in predictive ability between using "old" data as the training set and "new" data as the validation set and vice-versa. Comparing the additive genetic variance estimates for milk protein fractions between the FTIR predicted and HPLC measures, we found reductions of -19.7% for milk protein fractions expressed in g/L, and -21.19% expressed as % N. Although we found reductions in the heritability estimates, they were small, with values ranging from -1.9 to -7.25% for g/L, and -1.6 to -7.9% for % N. The posterior distributions of the additive genetic correlations (ra) between the FTIR predictions and the laboratory measurements were generally high (>0.8), even when the milk protein fractions were expressed as % N. Our results show the potential of using FTIR predictions in breeding programs as indicator traits for the selection of animals to enhance milk protein fraction contents. We expect acceptable responses to selection due to the high genetic correlations between HPLC measurements and FTIR predictions.


Asunto(s)
Proteínas de la Leche , Leche , Femenino , Bovinos , Animales , Proteínas de la Leche/análisis , Leche/química , Reproducibilidad de los Resultados , Espectrofotometría Infrarroja/veterinaria , Caseínas/análisis , Espectroscopía Infrarroja por Transformada de Fourier/veterinaria , Fenotipo
7.
Int J Mol Sci ; 24(13)2023 Jun 29.
Artículo en Inglés | MEDLINE | ID: mdl-37446034

RESUMEN

Tendinopathy, a prevalent overuse injury, lacks effective treatment options, leading to a significant impact on quality of life and socioeconomic burden. Mesenchymal stem/stromal cells (MSCs) and their secretome, including conditioned medium (CM) and extracellular vesicles (EVs), have shown promise in tissue regeneration and immunomodulation. However, it remains unclear which components of the secretome contribute to their therapeutic effects. This study aimed to compare the efficacy of CM, EVs, and the soluble protein fraction (PF) in treating inflamed tenocytes. CM exhibited the highest protein and particle concentrations, followed by PF and EVs. Inflammation significantly altered gene expression in tenocytes, with CM showing the most distinct separation from the inflamed control group. Treatment with CM resulted in the most significant differential gene expression, with both upregulated and downregulated genes related to inflammation and tissue regeneration. EV treatment also demonstrated a therapeutic effect, albeit to a lesser extent. These findings suggest that CM holds superior therapeutic efficacy compared with its EV fraction alone, emphasizing the importance of the complete secretome in tendon injury treatment.


Asunto(s)
Vesículas Extracelulares , Células Madre Mesenquimatosas , Humanos , Medios de Cultivo Condicionados/farmacología , Medios de Cultivo Condicionados/metabolismo , Tenocitos/metabolismo , Calidad de Vida , Vesículas Extracelulares/metabolismo , Inflamación/metabolismo , Proteínas/metabolismo , Células Madre Mesenquimatosas/metabolismo
8.
J Anim Breed Genet ; 139(5): 517-529, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35485246

RESUMEN

Considerable resources are required to routinely measure detailed milk compositional traits. Hence, an insufficient volume of phenotypic data can hinder genetic progress in these traits within dairy cow breeding programmes. The objective of the present study was to quantify the opportunities for breeding for improved milk protein and free amino acid (FAA) composition by exploiting mid-infrared spectroscopy (MIRS) predictions routinely recorded from milk samples. Genetic parameters for protein fractions and FAA composition were estimated using 134,546 test-day records from 16,166 lactations on 9,572 cows using linear mixed models. Heritability of MIRS-predicted protein fractions ranged from 0.19 (α-lactalbumin) to 0.55 (ß-lactoglobulin A), while heritability of MIRS-predicted FAA ranged from 0.08 for glycine to 0.29 for glutamic acid. Genetic correlations among the MIRS-predicted FAA were moderate to strong ranging from -0.44 (aspartic acid and lysine) to 0.97 (glutamic acid and total FAA). Adjustment of the genetic correlations for the genetic merit of 24-h milk yield did not greatly affect the correlations. Results from the current study highlight the presence of exploitable genetic variation for both protein fractions and FAA in dairy cow milk. Besides, the direction of genetic correlations reveals that breeding programmes directly selecting for greater milk protein concentration carry with them favourable improvement in casein and whey fractions.


Asunto(s)
Aminoácidos , Ácido Glutámico , Aminoácidos/análisis , Animales , Bovinos/genética , Femenino , Ácido Glutámico/análisis , Ácido Glutámico/genética , Ácido Glutámico/metabolismo , Lactancia/genética , Leche/química , Proteínas de la Leche/genética , Proteínas de la Leche/metabolismo , Fenotipo
9.
Stomatologiia (Mosk) ; 100(6): 108-114, 2021.
Artículo en Ruso | MEDLINE | ID: mdl-34953198

RESUMEN

Sjogren's disease (BS) is a systemic autoimmune disease, the main symptoms of which are associated with dry eyes, mouth, skin, respiratory and gastrointestinal tract, etc., but there are a large number of extra-vascular symptoms that can accompany this disease: weakness, swelling of the lymph nodes, respiratory failure, pain and swelling of the joints, rash, dysphagia, gastro-esophageal reflux, lymphoma, etc. In addition, the etiology of this disease remains unknown, and the pathogenesis is partially studied. The diagnosis of Sjogren's disease is complicated by a variety of clinical manifestations, as well as subacute and chronic variants of the course of the disease, in addition, the diagnostic criteria are periodically changed. It is recommended to adhere to the domestic diagnostic criteria considering the presence of laboratory signs of an autoimmune disease. The article presents a clinical observation of a patient with the development of the main symptoms and signs of BS and autoimmune hepatitis for 20 years. Recurrent mumps or chronic bilateral enlargement of the parotid salivary glands may be the first symptom of Sjogren's disease, which is recommended to extend the examination in order to identify other, sometimes latent signs of the disease.


Asunto(s)
Síndrome de Sjögren , Humanos , Hígado , Síndrome de Sjögren/complicaciones , Síndrome de Sjögren/diagnóstico
10.
Molecules ; 25(8)2020 Apr 14.
Artículo en Inglés | MEDLINE | ID: mdl-32295155

RESUMEN

Algae are employed commonly in cosmetics, food and pharmaceuticals, as well as in feed production and biorefinery processes. In this study, post-fermentation leachate from a biogas plant which exploits stillage and maize silage was utilized as a culture medium for Chlorella vulgaris. The content of polyphenols in hydrophilic extracts of the Chlorella vulgaris biomass was determined, and the extracts were evaluated for their antioxidant activity (DPPH assay), antibacterial activity (against Escherichia coli, Lactobacillus plantarum, Staphylococcus aureus, Staphylococcus epidermidis) and antifungal activity (against Aspergillus niger, Candida albicans, Saccharomyces cerevisiae). The use of the post-fermentation leachate was not found to affect the biological activity of the microalgae. The aqueous extract of Chlorella vulgaris biomass was also observed to exhibit activity against nematodes. The results of this study suggest that Chlorella vulgaris biomass cultured on post-fermentation leachate from a biogas plant can be successfully employed as a source of natural antioxidants, food supplements, feed, natural antibacterial and antifungal compounds, as well as in natural methods of plant protection.


Asunto(s)
Chlorella vulgaris/química , Fermentación , Microalgas/química , Extractos Vegetales/química , Extractos Vegetales/farmacología , Zea mays/química , Antioxidantes/química , Antioxidantes/farmacología , Interacciones Hidrofóbicas e Hidrofílicas , Ultrafiltración
11.
J Sci Food Agric ; 100(3): 1072-1079, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31650550

RESUMEN

BACKGROUND: Acid-induced hydrolysis of proteins has been used to improve the solubility and functional properties of various proteins, and could be a promising tool to facilitate the use of currently underutilized insoluble microalgae protein-rich fractions in food applications. However, the results of a prior study showed an unusual resistance of an insoluble microalgae protein-rich fraction to acid hydrolysis at room temperature. RESULTS: In the present study, the insoluble protein-rich fraction extracted from microalgae Chlorella prothothecoides was treated with 0.5 mol L-1 hydrochloric acid at 25, 45, 65 or 85 °C for 0-4 h. The results showed that hydrolysis of the fraction at 85 °C for 4 h led to decreases in the amount of insoluble protein-rich aggregates and the formation of fragments with a lower molecular weight, as well as an increase in protein solubility by approximately 40%. Nevertheless, some aggregated insoluble protein-rich particles remained, even after hydrolysis at 85 °C for 4 h. CONCLUSION: The higher temperature improved the efficiency of the acid hydrolysis of the insoluble protein fraction from microalgae Chlorella prothothecoides, which is highly acid-resistant. Overall, an erosion-based mechanism was suggested for the acid hydrolysis of insoluble microalgae protein fraction. © 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Asunto(s)
Chlorella/química , Proteínas/química , Fraccionamiento Químico , Calor , Ácido Clorhídrico/química , Hidrólisis , Microalgas/química , Peso Molecular , Proteínas/aislamiento & purificación , Solubilidad
12.
J Dairy Sci ; 101(12): 11108-11119, 2018 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-30316608

RESUMEN

The aim of this study was to investigate in Holstein cows the genetic basis of blood serum metabolites [i.e., total protein, albumin, globulin, albumin:globulin ratio (A:G), and blood ß-hydroxybutyrate (BHB)], a set of milk phenotypes related to udder health, milk quality technological characteristics, and genetic relationships among them. Samples of milk were collected from 498 Holstein cows belonging to 28 herds. All animal welfare and milk phenotypes were assessed using standard analytical methodology. A set of Bayesian univariate and bivariate animal models was implemented via Gibbs sampling, and statistical inference was based on the marginal posterior distributions of parameters of concern. We observed a small additive genetic influence for serum albumin concentrations, moderate heritability (≥0.20) for total proteins, globulins, and A:G, and high heritability (0.37) for blood BHB. Udder health traits (somatic cell score, milk lactose, and milk pH) showed low or moderate heritabilities (0.15-0.20), whereas variations in milk protein fraction concentrations were confirmed as mostly under genetic control (heritability: 0.21-0.71). The moderate and high heritabilities observed for milk coagulation properties and curd firming modeling parameters provided confirmation that genetic background exerts a strong influence on the cheese-making ability of milk, largely due to genetic polymorphisms in the major milk protein genes. Blood BHB showed strong negative genetic correlations with globulins (-0.619) but positive correlations with serum albumin (0.629) and A:G (0.717), which suggests that alterations in the serum protein pattern and BHB blood levels are likely to be genetically related. Strong relationships were found between albumin and fat percentages (-0.894), between globulin and αS2-CN (-0.610), and, to a lesser extent, between serum protein pattern and milk technological characteristics. Genetic relationships between blood BHB and traits related to udder health and milk quality and technological characteristics were mostly weak. This study provides evidence that there is exploitable additive genetic variation for traits related to animal health and welfare and throws light on the shared genetic basis of these traits and the phenotypes related to the quality and cheese-making ability of milk.


Asunto(s)
Ácido 3-Hidroxibutírico/sangre , Bovinos/genética , Queso/análisis , Variación Genética , Glándulas Mamarias Animales/metabolismo , Proteínas de la Leche/metabolismo , Animales , Bovinos/sangre , Femenino , Lactosa/análisis , Leche/química , Proteínas de la Leche/química , Proteínas de la Leche/genética , Fenotipo , Carácter Cuantitativo Heredable
13.
Molecules ; 23(2)2018 Feb 12.
Artículo en Inglés | MEDLINE | ID: mdl-29439533

RESUMEN

Some molecular and functional properties of albumin (83.6% protein), globulin (95.5% protein), glutelin (81.3% protein) as well as protein isolate (80.7% protein) from cashew nut were investigated. These proteins were subjected to molecular (circular dichroism, gel electrophoresis, scanning electron microscopy) and functional (solubility, emulsification, foaming, water/oil holding capacity) tests. Cashew nut proteins represent an abundant nutrient with well-balanced amino acid composition and could meet the requirements recommended by FAO/WHO. SDS-PAGE pattern indicated cashew nut proteins were mainly composed of a polypeptide with molecular weight (MW) of 53 kDa, which presented two bands with MW of 32 and 21 kDa under reducing conditions. The far-UV CD spectra indicated that cashew proteins were rich in ß-sheets. The surface hydrophobicity of the protein isolate was higher than that of the protein fractions. In pH 7.0, the solubility of protein fractions was above 70%, which was higher than protein isolate at any pH. Glutelin had the highest water/oil holding capacity and foaming properties. Protein isolate displayed better emulsifying properties than protein fractions. In summary, cashew nut kernel proteins have potential as valuable nutrition sources and could be used effectively in the food industry.


Asunto(s)
Albúminas/química , Anacardium/química , Proteínas en la Dieta/química , Globulinas/química , Glútenes/química , Nueces/química , Proteínas de Plantas/química , Albúminas/aislamiento & purificación , Proteínas en la Dieta/aislamiento & purificación , Electroforesis en Gel de Poliacrilamida , Análisis de los Alimentos , Globulinas/aislamiento & purificación , Glútenes/aislamiento & purificación , Concentración de Iones de Hidrógeno , Peso Molecular , Proteínas de Plantas/aislamiento & purificación , Conformación Proteica en Lámina beta , Solubilidad
14.
Biochim Biophys Acta Gen Subj ; 1861(3): 615-623, 2017 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-28063984

RESUMEN

BACKGROUND: Peptide and protein toxins are essential tools to dissect and probe the biology of their target receptors. Venoms target vital physiological processes to evoke pain. Snake venoms contain various factors with the ability to evoke, enhance and sustain pain sensation. While a number of venom-derived toxins were shown to directly target TRPV1 channels expressed on somatosensory nerve terminals to evoke pain response, such toxins were yet to be identified in snake venoms. METHODS: We screened Echis coloratus saw-scaled viper venom's protein fractions isolated by reversed phase HPLC for their ability to activate TRPV1 channels. To this end, we employed heterologous systems to analyze TRPV1 and NGF pathways by imaging and electrophysiology, combined with molecular biology, biochemical, and pharmacological tools. RESULTS: We identified TRPV1 activating proteins in the venom of Echis coloratus that produce a channel-dependent increase in intracellular calcium and outwardly rectifying currents in neurons and heterologous systems. Interestingly, channel activation was not mediated by any of its known toxin binding sites. Moreover, although NGF neurotropic activity was detected in this venom, TRPV1 activation was independent of NGF receptors. CONCLUSIONS: Echis coloratus venom contains proteins with the ability to directly activate TRPV1. This activity is independent of the NGF pathway and is not mediated by known TRPV1 toxins' binding sites. GENERAL SIGNIFICANCE: Our results could facilitate the discovery of new toxins targeting TRPV1 to enhance current understanding of this receptor activation mechanism. Furthermore, the findings of this study provide insight into the mechanism through which snakes' venom elicit pain.


Asunto(s)
Proteínas/metabolismo , Canales Catiónicos TRPV/metabolismo , Venenos de Víboras/metabolismo , Viperidae/metabolismo , Animales , Sitios de Unión/fisiología , Calcio/metabolismo , Línea Celular , Células HEK293 , Humanos , Factor de Crecimiento Nervioso/metabolismo , Neuronas/metabolismo , Receptores de Factor de Crecimiento Nervioso/metabolismo , Toxinas Biológicas/metabolismo
15.
J Dairy Sci ; 100(7): 5526-5540, 2017 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-28478002

RESUMEN

The objectives of this study were to estimate, for the Italian Simmental cattle population, genetic parameters for 92 traits and their infrared predictions (IP) and to investigate the genetic relationship between measured traits (MT) and IP. Data for milk fat fatty acid composition (n = 1,040), detailed protein composition (n = 3,337), lactoferrin (n = 558), pH (n = 3,438), coagulation properties (n = 3,266), curd yield and composition obtained by a micro-cheese making procedure (n = 1,177), and content of Ca, P, Mg, and K (n = 689) were obtained using reference laboratory analysis. Infrared prediction for all the investigated traits was performed using 143,198 spectra records belonging to 17,619 Italian Simmental cows. (Co)variance components for MT and their IP were estimated in a set of bivariate animal model REML analyses and genetic correlations between MT and IP were estimated using all IP obtained at the population level. A significant positive relationship was observed between the coefficient of determination of the infrared prediction models and the phenotypic and genetic variation of the IP. The decrease in the estimated genetic variance of IP compared with MT was on average 64%. For traits exhibiting calibration models with coefficients of determination in cross-validation (R2CV) greater than 0.9, the decrease in the genetic variance ranged from approximately 20 to 50%. Most traits (88 out of 92) exhibited lower heritability estimates for IP than for the corresponding MT. The estimated genetic correlations between IP and MT (ra) were in general very high. A positive relationship (r = 0.57) between R2CV of calibration models and the estimated ra has been detected. For calibration models exhibiting R2CV higher than 0.75, ra were greater than 0.9. The variability in the estimated correlations increased when R2CV decreased, and for calibration models of moderate predictive ability, estimates of ra ranged from 0.2 to 1. Genetic parameter estimates suggested that IP can be used as indicator traits in breeding programs for the enhancement of fine composition and technological properties of milk. The genetic gain achievable selecting for IP is expected to be high for fatty acid composition, minerals, and for technological properties of milk, whereas it will be low for casein and whey protein composition and for the content of lactoferrin.


Asunto(s)
Glucolípidos/química , Glicoproteínas/química , Proteínas de la Leche/química , Leche/química , Fenotipo , Animales , Cruzamiento , Caseínas , Bovinos , Queso , Femenino , Variación Genética , Glucolípidos/genética , Glicoproteínas/genética , Italia , Gotas Lipídicas , Proteínas de la Leche/genética
16.
J Dairy Sci ; 100(9): 7306-7319, 2017 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-28647337

RESUMEN

The objective of this study was to compare the prediction accuracy of 92 infrared prediction equations obtained by different statistical approaches. The predicted traits included fatty acid composition (n = 1,040); detailed protein composition (n = 1,137); lactoferrin (n = 558); pH and coagulation properties (n = 1,296); curd yield and composition obtained by a micro-cheese making procedure (n = 1,177); and Ca, P, Mg, and K contents (n = 689). The statistical methods used to develop the prediction equations were partial least squares regression (PLSR), Bayesian ridge regression, Bayes A, Bayes B, Bayes C, and Bayesian least absolute shrinkage and selection operator. Model performances were assessed, for each trait and model, in training and validation sets over 10 replicates. In validation sets, Bayesian regression models performed significantly better than PLSR for the prediction of 33 out of 92 traits, especially fatty acids, whereas they yielded a significantly lower prediction accuracy than PLSR in the prediction of 8 traits: the percentage of C18:1n-7 trans-9 in fat; the content of unglycosylated κ-casein and its percentage in protein; the content of α-lactalbumin; the percentage of αS2-casein in protein; and the contents of Ca, P, and Mg. Even though Bayesian methods produced a significant enhancement of model accuracy in many traits compared with PLSR, most variations in the coefficient of determination in validation sets were smaller than 1 percentage point. Over traits, the highest predictive ability was obtained by Bayes C even though most of the significant differences in accuracy between Bayesian regression models were negligible.


Asunto(s)
Teorema de Bayes , Análisis de los Mínimos Cuadrados , Fenotipo , Espectroscopía Infrarroja Corta/veterinaria , Animales , Caseínas/química , Queso , Industria Lechera/estadística & datos numéricos , Espectroscopía Infrarroja Corta/métodos
17.
J Dairy Sci ; 100(3): 2057-2067, 2017 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-28109603

RESUMEN

The aim of this study was to compare the common method of exploiting infrared spectral data in animal breeding; that is, estimating the breeding values for the traits predicted by infrared spectroscopy, and an alternative approach based on the direct use of spectral information (direct prediction, DP) to predict the estimated breeding values (EBV). Traits were pH, milk coagulation properties, contents of the main casein and whey protein fractions, cheese yield measured by micro-cheese making, lactoferrin, Ca, and fat composition. For the DP method, the number of spectral variables was reduced by principal components analysis to 8 latent traits that explained 99% of the original spectral variation. Restricted maximum likelihood was used to estimate variance components of the latent traits. (Co)variance components of the original spectral traits were obtained by back-transformation and EBV of all derived milk traits were then predicted as traits correlated with the genetic information of the spectra. The rank correlation between the EBV obtained for the infrared-predicted traits and those obtained from the DP method was variable across traits. Rank correlations ranged from 0.07 (for the content of saturated fatty acids expressed as g/100 g of fat) to 0.96 (for dry matter cheese yield, %) and, for most traits, was <0.5. This result can be explained by the nature of the principal components analysis: it does not take into account the covariance between the spectral variables and the reference traits but produces latent traits that maximize the spectral variance explained. Thus, the direct approach is more likely to be effective for traits more related to the main sources of spectral variation (i.e., protein and fat). More research is required to study spectral genetic variation and to determine the best way to choose spectral regions and the type and number of considered latent traits for potential applications.


Asunto(s)
Análisis de Fourier , Leche/química , Animales , Cruzamiento , Queso , Espectroscopía Infrarroja por Transformada de Fourier
18.
J Food Sci Technol ; 54(7): 1866-1872, 2017 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-28720942

RESUMEN

In present study, three varieties (G 80, Ageta 112 and HG 365) of guar bean (Cyamopsis tetragonoloba) were analysed for proximate analysis and were processed by different methods (dehusking, soaking, autoclaving, extrusion and germination) to reduce its antinutritional factors. Processed guar flours were studied for antinutritional factors (tannins, phytic acid and polyphenols) and protein fractions. The highest protein, ash and polyphenols contents were observed in Ageta 112. G 80 contained the lowest tannin and phytic acid content. High temperature treatments (i.e. autoclaving at 110 °C/10 min, 120 psi and extrusion-Clextral, Twin screw extruder) were found to be most effective in reducing the tannin and polyphenol content. More than 90% reduction in tannins was observed with high temperature treatments in HG 365. Phytic acid fraction increased slightly on soaking, however, extensive reduction was observed with other treatments. Globulins formed the major protein fraction in guar bean and various processing treatments significantly affected the protein fractions. Autoclaving was observed to be the best treatment to reduce antinutritional factors in guar bean and thereafter, its utilization in food.

19.
J Dairy Sci ; 99(3): 1853-1858, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26774721

RESUMEN

The objective of this study was to investigate the ability of mid-infrared spectroscopy (MIRS) to predict protein fraction contents of bovine milk samples by applying uninformative variable elimination (UVE) procedure to select the most informative wavenumber variables before partial least squares (PLS) analysis. Reference values (n=114) of protein fractions were measured using reversed-phase HPLC and spectra were acquired through MilkoScan FT6000 (Foss Electric A/S, Hillerød, Denmark). Prediction models were built using the full data set and tested with a leave-one-out cross-validation. Compared with MIRS models developed using standard PLS, the UVE procedure reduced the number of wavenumber variables to be analyzed through PLS regression and improved the accuracy of prediction by 6.0 to 66.7%. Good predictions were obtained for total protein, total casein (CN), and α-CN, which included αS1- and αS2-CN; moderately accurate predictions were observed for κ-CN and total whey protein; and unsatisfactory results were obtained for ß-CN, α-lactalbumin, and ß-lactoglobulin. Results indicated that UVE combined with PLS is a valid approach to enhance the accuracy of MIRS prediction models for milk protein fractions.


Asunto(s)
Caseínas/análisis , Análisis de los Alimentos/métodos , Proteínas de la Leche/análisis , Espectrofotometría Infrarroja/métodos , Animales , Bovinos , Cromatografía Líquida de Alta Presión , Dinamarca , Lactalbúmina/análisis , Lactoglobulinas , Análisis de los Mínimos Cuadrados , Modelos Teóricos , Valor Predictivo de las Pruebas , Proteína de Suero de Leche
20.
Cent Eur J Immunol ; 40(4): 400-10, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26862303

RESUMEN

The aim of the study was to evaluate analgesic activity ("hot plate" test), anti-inflammatory activity (carrageenan-induced paw edema) and locomotor activity in rats under the influence of three fractions of Chelidonium majus herb extract: full water extract (FWE), protein enriched fraction (PEF), and non-protein fraction (NPF). Effects of the fractions on the level of chosen cytokines and their mRNA levels were also assessed using lipopolysaccharide (LPS) administration as a proinflammatory cue. All fractions and diclofenac did not affect the locomotor activity of rats in comparison with the control group. FWE and PEF three hours after administration showed statistically significant analgesic activities comparable to morphine (p < 0.05). A slight reduction in rat paw edema was observed after three (comparable with diclofenac) and six hours in the NPF group. FWE revealed a statistically significant pro-inflammatory effect after three hours in comparison with the control group. Peripheral IL-1 and IL-4 cytokine concentrations were reduced under FWE and NPF, PEF fractions. The combination of FWE, PEF and NPF together with LPS showed only the effects of LPS. We suggest that protein enriched fraction (PEF) produced centrally mediated (morphine-like) analgesic action, whereas the anti-inflammatory potential was shown only after LPS-induced inflammation. The precise mechanisms involved in the production of anti-nociceptive and anti-inflammatory responses of studied fractions are not completely understood, but they may be caused rather by the presence of protein more than alkaloids-enriched fraction. This fraction of the extract could be used as an alternative therapy for the prevention of inflammatory-related diseases in the future, but further studies are needed.

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