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1.
Int J Food Sci ; 2022: 4289059, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36245563

RESUMO

Deproteinized sunflower meal (DSM) was obtained as waste from ethanol-treated sunflower meal after alkaline extraction of proteins. The study aimed at biochemically and functionally characterizing the material concerning its potential practical application and valuability. The DSM consisted mainly of proteins (19.88%) and dietary fibers (61.06%) the majority of which were insoluble (53.09%). Cellulose (30.87%) and lignin (21.79%) were the most contributing compounds to the total amount of dietary fibers. The DSM contained Fe (133.29 mg/kg), Zn (201.56 mg/kg), and Cu (31.87 mg/kg). The analyses defined the DSM as a fiber concentrate with relatively high thermal stability. The distraction of the material began at 170°Ð¡ with a maximum speed at 277°Ð¡. The highest water absorption capacity (WAC) of the DSM was observed at pH 6 and 7 (approximately 8 g H2O/g sample) under all studied conditions including pH from 3 to 10 and three levels of NaCl concentrations (0.00 M, 0.03 M, and 0.25 M). At pH 7, increasing temperature from 20°C to 60°C increased the WAC of the DSM from 8.13 g H2O/g sample to 9.80 g H2O/g sample, respectively. Further increase in the temperature diminished the WAC of the DSM. At pH 6, the increase in temperature did not influence positively the WAC of the DSM. The study demonstrated the potential of the DSM, a waste obtained from the protein isolation process, as a valuable ingredient/additive in the food industry.

2.
Foods ; 11(4)2022 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-35206016

RESUMO

Rapeseed meal is a by-product of the oil-producing industry with a currently underestimated application. Two protein isolates, PI2.5-8.5 or PI10.5-2.5, were obtained from industrial rapeseed meal after treatment with an aqueous ethanol solution. The alkaline-extracted proteins were sequentially precipitated by two different modes, from pH 10.5 to 2.5, and vice versa, from 2.5 to 8.5, with a step of 1 pH unit. The preparation approach influenced both the functional and antioxidant properties of the isolates. The PI10.5-2.5 exhibited higher water and oil absorption capacities than PI2.5-8.5, reaching 2.68 g H2O/g sample and 2.36 g oil/g sample, respectively. The emulsion stability of the PI2.5-8.5, evaluated after heating at 80 °C, was either 100% or close to 100% for all pH values studied (from 2 to 10), except for pH 6 where it reached 93.87%. For the PI10.5-2.5, decreases in the emulsion stability were observed at pH 8 (85.71%) and pH 10 (53.15%). In the entire concentration range, the PI10.5-2.5 exhibited a higher scavenging ability on 2,2-diphenyl-1-picryl hydrazyl (DPPH) and hydroxyl radicals than PI2.5-8.5 as evaluated by DPPH and 2-deoxyribose assays, respectively. At the highest concentration studied, 1.0%, the neutralization of DPPH radicals by PI10.5-2 reached half of that exhibited by synthetic antioxidant butylhydroxytoluene (82.65%). At the same concentration, the inhibition of hydroxyl radicals by PI10.5-2 (71.25%) was close to that achieved by mannitol (75.62%), which was used as a positive control. Established antioxidant capacities add value to the protein isolates that can thus be used as both emulsifiers and antioxidants.

3.
Foods ; 9(6)2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32492803

RESUMO

The solubility of plant protein isolates is a key determinant of their potential application. Two protein isolates (PI) from ethanol-treated industrial rapeseed meal, PI10.5-2.5 and PI2.5-8.5, were prepared by sequential isoelectric precipitation of alkali-extracted proteins (pH 12) starting from pH 10.5 to 2.5 or from pH 2.5 to 8.5, respectively. Biochemical analyses revealed that PI2.5-8.5 contained a higher amount of crude protein (72.84%) than PI10.5-2.5 (68.67%). In the same protein isolate, the level of total phenols (0.71%) was almost two-fold higher than that in PI10.5-2.5 (0.42%). No glucosinolates were established in both protein isolates. SDS-PAGE analysis demonstrated that PI10.5-2.5 contained 10 to 15 kDa protein fractions in a relatively higher amount, while PI2.5-8.5 was enriched in 18 to 29 kDa protein fractions. PI10.5-2.5 exhibited high solubility, varying from 41.74% at pH 4.5 to 65.13% at pH 6.5, while PI2.5-8.5 was almost two-fold less soluble under the same conditions. Up to pH 5.5, the addition of NaCl at 0.03 and 0.25 M diminished the solubility of PI2.5-8.5, while the solubility of PI10.5-2.5 was increased. The supplementation of PI10.5-2.5 with 0.25 M NaCl enhanced the protein solubility to 56.11% at pH 4.5 and 94.26% at pH 6.5. The addition of 0.03 M NaCl also increased the solubility of this protein isolate but to a lower extent. Overall, the approach for sequential precipitation of proteins influenced the biochemical characteristics, protein fractional profile and solubility of prepared protein isolates.

4.
Ecotoxicol Environ Saf ; 194: 110413, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32163775

RESUMO

Polymetal dust is a common industrial pollutant. While the use of remediation filters and equipment in lead smelters has reduced pollutant emission, surrounding areas remain contaminated due to the long-term transfer of heavy metals along the food chain. Here we assess the mutagenic potential of the lead-zinc smelter near Plovdiv (Bulgaria) situated in an area that has been contaminated with heavy metals for 60 years. We aimed to evaluate the genomic response of the yellow-necked mouse (A. flavicollis), a biomonitor species, in three sampling sites along the pollution gradient. Mice from Strandzha Natural Park were used as a negative control. The bioaccumulation rate of two non-essential heavy metals, lead (Pb) and cadmium (Cd), in liver tissues was determined by atomic absorption spectroscopy. Genetic alterations attributable to chronic exposure to trace levels of heavy metals were assessed in different blood cell populations using two independent methods: a micronucleus test was applied to evaluate the clastogenic and aneugenic alterations in erythrocytes, while a comet assay was used to assess DNA instability, as evidenced by single- and double-stranded breaks and alkali-labile sites, in leucocytes. We observed elevated levels of Pb and Cd in livers derived from mice from the impacted area: the mean Pb concentration (21.38 ± 8.77 µg/g) was two-fold higher than the lowest-observed-adverse-effect levels (LOAELs), while the mean Cd concentration (13.95 ± 9.79 µg/g) was extremely close to these levels. The mean levels of Pb and Cd in livers derived from mice from the impacted area were 31-fold and 63-fold higher, respectively, than the levels measured in mice from the control area. The mean frequency of micronuclei was significantly higher (four-fold) than that observed in the control animals. Furthermore, parameters measured by the comet assay, % tail DNA, tail length and tail moment, were significantly higher in the impact area, indicating the degree of genetic instability caused by exposure to heavy metals. In conclusion, this study shows that despite the reported reduction in lead and cadmium emissions in Bulgaria in recent years, A. flavicollis individuals inhabiting areas subject to long-term contamination exhibit significant signs of DNA damage.


Assuntos
Monitoramento Biológico/métodos , Cádmio/toxicidade , Dano ao DNA , Poluentes Ambientais/toxicidade , Instabilidade Genômica/efeitos dos fármacos , Chumbo/toxicidade , Animais , Bulgária , Cádmio/análise , Ensaio Cometa , Relação Dose-Resposta a Droga , Poeira/análise , Poluentes Ambientais/análise , Chumbo/análise , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Camundongos , Testes para Micronúcleos
5.
J Food Sci Technol ; 56(6): 3090-3098, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31205363

RESUMO

A protein isolate (ERPI) was prepared from ethanol-treated rapeseed meal and used as a stabilizing agent in sunflower and rapeseed oil-in-water emulsions. The aim of the current study was to explore the influence of protein and oil concentrations on initial stability of sunflower and rapeseed oil-in-water emulsions by evaluating Gibbs free energy (ΔG) and particle size distribution. The 7-day dynamics of emulsion stability was investigated by turbidity measurement as well. A 32 factorial design was applied to assess the significance of oil (5%, 10% and 15% w/w) and ERPI protein (0.25%, 0.5% and 1.0% w/w) addition on stability of the emulsions. The results demonstrated that the increase of oil concentrations from 5 to 15% positively influenced the initial stability of sunflower and rapeseed oil-in-water emulsions. In both oil types, ERPI protein supplementation at all levels resulted in significant differences in the stability of 5% and 10% oil emulsions but did not alter the initial stability of the emulsions prepared with either 15% sunflower or rapeseed oil. With a few exceptions, there was a good agreement between Gibbs free energy data and microstructural profiles of the emulsions. Overall, emulsions with all sunflower oil concentrations and 1.0% ERPI protein exhibited better initial and a 7-day stability dynamics compared to all rapeseed oil-based emulsions. The study demonstrated the potential of ethanol-treated rapeseed meal protein isolate to serve as an emulsifying agent in sunflower and rapeseed oil containing emulsions.

6.
J Food Sci Technol ; 55(9): 3792-3798, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30150839

RESUMO

The use of the rapeseed meal as a source for preparation of protein-rich ingredients for the food industry is an alternative to the current limited application as a feed additive. The aim of this study was to evaluate foaming properties of an acid-soluble protein-rich ingredient (ASP) obtained from industrial rapeseed meal as a co-product of a protein isolate. Foam capacity and stability over a period of 60 min were evaluated by using volumetric and image analyzing methods. The influence of NaCl at two boundary concentrations (0.03 and 0.25 M) was studied over a pH range from 2 to 10. The ASP exhibited high foamability (> 90%), not influenced by pH or salt addition. In contrast, foam stability, measured over a 60 min period, was pH and NaCl dependent. By the end of the observation period, the addition of 0.25 M NaCl reduced the foam volume by more than 70% at all pH values. After 30 min at pH values 4, 6 and 8, which are the most common for food products, the foams without NaCl retained 51, 38 and 41% of the initial foam volume, respectively. The results were in agreement with image analysis observations where microstructure of the foams with NaCl was more heterogeneous than that of the foams without salt addition. The high foamability and relatively high foam stability at pH from 4 to 8 without NaCl addition shows that ASP could be a potential alternative to plant proteins currently used as foaming agents in the food industry.

7.
Food Technol Biotechnol ; 55(3): 420-428, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29089856

RESUMO

The utilization of industrial sunflower meal to produce protein-rich products for the food industry is an alternative approach for better and more efficient use of this agricultural by-product. Sunflower meal proteins possess specific functional properties, which however need improvement to broaden their potential as supplements for delivering high--quality products for human nutrition. The aim of the study is to evaluate the combined influence of low-degree pepsin hydrolysis and transglutaminase (TG) modification on industrial sunflower meal protein isolate functionality at pH=2 to 10. Three TG-modified pepsin hydrolysates with the degree of hydrolysis of 0.48, 0.71 and 1.72% were produced and named TG-PH1, TG-PH2 and TG-PH3, respectively. All three TG-modified pepsin hydrolysates exhibited improved solubility at pH between 3.5 and 5.5 as the highest was observed of TG-PH3 at protein isoelectric point (pI=4.5). Sunflower meal protein isolate and TG-modified sunflower meal protein isolate had greater solubility than the three TG-modified hydrolysates at pH<3 and >7. Significant improvement of foam making capacity (p<0.05) was achieved with all three TG-modified pepsin hydrolysates in the entire pH area studied. Pepsin hydrolysis of the protein isolate with the three degrees of hydrolysis did not improve foam stability. Improved thermal stability was observed with TG-PH3 up to 80 °C compared to the protein isolate (pH=7). At 90 °C, TG modification of the protein isolate alone resulted in the highest thermal stability. Pepsin hydrolysis followed by a treatment with TG could be used to produce sunflower protein isolates with improved solubility, foam making capacity and thermal stability for use in the food industry.

8.
Osteoarthritis Cartilage ; 25(9): 1522-1530, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28583899

RESUMO

OBJECTIVE: Statins are widely used drugs for cholesterol lowering, which were recently found to counteract the effects of aberrant fibroblast growth factor receptor (FGFR3) signaling in cell and animal models of FGFR3-related chondrodysplasia. This opened an intriguing therapeutic possibility for human dwarfing conditions caused by gain-of-function mutations in FGFR3, although the mechanism of statin action on FGFR3 remains unclear. Here, we determine the effect of statins on FGFR signaling in chondrocytes. DESIGN: Cultured chondrocyte cell lines, mouse embryonic tibia cultures and limb bud micromasses were treated with FGF2 to activate FGFR signaling. The effects of atorvastatin, fluvastatin, lovastatin and pravastatin on FGFR3 protein stability and on FGFR-mediated chondrocyte growth-arrest, loss of extracellular matrix (ECM), induction of premature senescence and hypertrophic differentiation were evaluated. RESULTS: Statins did not alter the level of FGFR3 protein expression nor produce any effect on FGFR-mediated inhibition of chondrocyte proliferation and hypertrophic differentiation in cultured chondrocyte cell lines, mouse tibia cultures or limb bud micromasses. CONCLUSION: We conclude that statins do not inhibit the FGFR signaling in chondrocytes. Therefore the statin-mediated rescue of FGFR3-related chondrodysplasia, described before, is likely not intrinsic to the growth plate cartilage.


Assuntos
Condrócitos/efeitos dos fármacos , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Receptor Tipo 3 de Fator de Crescimento de Fibroblastos/antagonistas & inibidores , Animais , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Células Cultivadas , Condrócitos/metabolismo , Condrogênese/efeitos dos fármacos , Humanos , Botões de Extremidades/efeitos dos fármacos , Botões de Extremidades/metabolismo , Camundongos , Ratos , Receptor Tipo 3 de Fator de Crescimento de Fibroblastos/metabolismo , Transdução de Sinais/efeitos dos fármacos , Tíbia/efeitos dos fármacos , Tíbia/embriologia , Tíbia/crescimento & desenvolvimento , Técnicas de Cultura de Tecidos
9.
J Environ Sci Health B ; 51(4): 230-5, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26786395

RESUMO

Organic poultry is an alternative to conventional poultry which is rapidly developing as a response to customers' demand for better food and a cleaner environment. Although organic poultry manure can partially be utilized by organic horticultural producers, litter accumulation as well as excessive nitrogen still remains a challenge to maintain environment pureness, animal, and human health. Compared to conventional poultry, diet formulation without nitrogen overloading in organic poultry is even more complicated due to specific standards and regulations which limit the application of some supplements and imposes specific criteria to the ingredients in use. This is especially valid for methionine provision which supplementation as a crystalline form is only temporarily allowed. This review is focused on the utilization of various protein sources in the preparation of a diet composed of 100% organic ingredients which meet the avian physiology need for methionine, while avoiding protein overload. The potential to use unconventional protein sources such as invertebrates and microbial proteins to achieve optimal amino acid provision is also discussed.


Assuntos
Ração Animal , Fenômenos Fisiológicos da Nutrição Animal/fisiologia , Nitrogênio/metabolismo , Agricultura Orgânica/métodos , Aves Domésticas/fisiologia , Animais , Suplementos Nutricionais , Esterco , Metionina/farmacologia , Proteínas de Vegetais Comestíveis/administração & dosagem , Aves Domésticas/metabolismo
10.
Bone ; 76: 97-106, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25845979

RESUMO

The Myb locus encodes the c-Myb transcription factor involved in controlling a broad variety of cellular processes. Recently, it has been shown that c-Myb may play a specific role in hard tissue formation; however, all of these results were gathered from an analysis of intramembranous ossification. To investigate a possible role of c-Myb in endochondral ossification, we carried out our study on the long bones of mouse limbs during embryonic development. Firstly, the c-myb expression pattern was analyzed by in situ hybridization during endochondral ossification of long bones. c-myb positive areas were found in proliferating as well as hypertrophic zones of the growth plate. At early embryonic stages, localized expression was also observed in the perichondrium and interdigital areas. The c-Myb protein was found in proliferating chondrocytes and in the perichondrium of the forelimb bones (E14.5-E17.5). Furthermore, protein was detected in pre-hypertrophic as well as hypertrophic chondrocytes. Gain-of-function and loss-of-function approaches were used to test the effect of altered c-myb expression on chondrogenesis in micromass cultures established from forelimb buds of mouse embryos. A loss-of-function approach using c-myb specific siRNA decreased nodule formation, as well as downregulated the level of Sox9 expression, a major marker of chondrogenesis. Transient c-myb overexpression markedly increased the formation of cartilage nodules and the production of extracellular matrix as detected by intense staining with Alcian blue. Moreover, the expression of early chondrogenic genes such as Sox9, Col2a1 and activity of a Col2-LUC reporter were increased in the cells overexpressing c-myb while late chondrogenic markers such as Col10a1 and Mmp13 were not significantly changed or were downregulated. Taken together, the results of this study demonstrate that the c-Myb transcription factor is involved in the regulation and promotion of endochondral bone formation.


Assuntos
Condrogênese/fisiologia , Proteínas Proto-Oncogênicas c-myb/fisiologia , Animais , Biomarcadores/metabolismo , Diferenciação Celular , Extremidades/embriologia , Inativação Gênica , Hibridização In Situ , Camundongos , Proteínas Proto-Oncogênicas c-myb/genética
11.
Folia Biol (Praha) ; 57(4): 162-9, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21978758

RESUMO

Mouse embryonic carcinoma cells (P19 line) were studied for both their survival and developmental potential in the intact cerebellum of B6CBA mice. The P19 cells were cultured and labelled with green fluorescent protein using transfection. Cells were used for transplantation either in the undifferentiated stage or after 3 days of neurodifferentiation induced by retinoic acid. The intracerebellar application was performed in 43 mice: group A (N = 21) received neuroprogenitors and group B (N = 22) received undifferentiated cells. The morphology of transplanted cells within the context of the surrounding cerebellar tissue was evaluated after 3 weeks. Naive P19 cells engrafted and survived in the cerebellum of 7 of the 22 adult mice (survival rate 31.8 %). Neuroprogenitors survived in 13 of the 21 mice (survival rate was 61.9 %). Since the cut-off is P < 0.05, the difference is not statistically significant (P = 0.069). An expansive appearance of the graft was significantly more frequent (P = 0.0047) in naive P19 cells than in neuroprogenitors. In mice in which the grafts did not survive, no marks of grafted cells or only fluorescing detritus were found. In conclusion, this is the first study to track the fate and morphology of embryonic carcinoma cells transplanted into the cerebellum, confirming that neuroprogenitors derived from embryonic carcinoma cells can settle in the host tissue and differentiate according to the surrounding conditions. With further validation, the embryonic carcinoma cells could become a valuable model with which to study the impact of cell therapy on neurodegenerative diseases.


Assuntos
Sobrevivência Celular/fisiologia , Cerebelo/citologia , Neurônios/citologia , Células-Tronco/citologia , Animais , Western Blotting , Diferenciação Celular/fisiologia , Linhagem Celular Tumoral , Transplante de Células , Técnica Indireta de Fluorescência para Anticorpo , Camundongos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transplante de Células-Tronco
12.
Folia Biol (Praha) ; 56(4): 165-72, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20974049

RESUMO

The leukaemia inhibitory factor is a cytokine that exhibits pleiotropic activities in a wide range of cell types. There are evidences that leukaemia inhibitory factor-regulated signalling pathways are involved in cardiomyogesis and maintenance of cardiomyocytes. In the present work we studied the effect of leukaemia inhibitory factor on cardiomyogenesis of embryonic stem cells together with the role of serum-born factors. We showed that leukaemia inhibitory factor had an inhibitory effect during both the induction and progression phases of cardiomyogenesis of embryonic stem cells. The leukaemia inhibitory factor-mediated inhibition of cardiomyogenesis was abolished by inhibitors of STAT3 activity. These results suggest that leukaemia inhibitory factor- activated STAT3 is responsible for the inhibition of cardiomyogenesis in embryonic stem cells.


Assuntos
Diferenciação Celular , Células-Tronco Embrionárias/citologia , Fator Inibidor de Leucemia/fisiologia , Miócitos Cardíacos/citologia , Fator de Transcrição STAT3/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Embrião de Mamíferos , Indução Embrionária/efeitos dos fármacos , Células-Tronco Embrionárias/efeitos dos fármacos , Células-Tronco Embrionárias/metabolismo , Células-Tronco Embrionárias/fisiologia , Coração/embriologia , Fator Inibidor de Leucemia/metabolismo , Camundongos , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/fisiologia , Fator de Transcrição STAT3/farmacologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia
13.
J Environ Sci Health B ; 45(2): 162-6, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20390946

RESUMO

The presence of Maillard reaction products (MRP) in foods and food components is due to the non-enzymatic reaction between protein and carbohydrate residues triggered by thermal steps during food processing. The objective of this study was to assess the effect of MRPs and increasing lysine concentrations on S. Typhimurium growth and the expression of cadA which may be an indirect determinant of Salmonella virulence response. Variations in lysine concentrations (from 0 to 0.5 mM) did not exert any effect either on the final optical density after 6-hour incubation or the growth rates of S. Typhimurium in media containing MRPs. In contrast to the reduced final absorbancy of the bacterial cultures grown with histidine and arginine MRPs supplementations (0.1%), growth rates, in general, remained unaltered by all MRPs at each lysine concentration when compared to the control (M9 pH 5.8, no MRPs added). The induction levels of cadA in media containing 0.1% MRPs were close to cadA induction in the reference media (M9, pH 5.8 and no MRPs) and did not exceed the corresponding values by more than approximately 30%. Although the observed negligible induction of cadA under these conditions complies with the concept of its potential "anti-virulence" function, additional studies involving various concentrations and more refined MRPs are needed.


Assuntos
Carboxiliases/genética , Genes Bacterianos , Reação de Maillard , Salmonella typhimurium , Arginina/metabolismo , Carboxiliases/metabolismo , Manipulação de Alimentos , Microbiologia de Alimentos , Regulação Bacteriana da Expressão Gênica , Regulação Enzimológica da Expressão Gênica , Histidina/metabolismo , Concentração de Íons de Hidrogênio , Lisina/metabolismo , Salmonella typhimurium/genética , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/metabolismo
14.
J Sci Food Agric ; 90(5): 870-6, 2010 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-20355124

RESUMO

BACKGROUND: Due to their low solubility in water, oil-based bioactive compounds require dispersion in a surface-active agent or appropriate solvents to ensure maximum contact with microorganisms. These combinations, however, may change their physical and/or chemical characteristics and consequently alter the desired functionality. The objective of this study was to determine the impact of selected dispersing agents, ethanol, dimethyl sulfoxide (DMSO), and Tween-80, on cold-pressed terpeneless (CPT) Valencia orange oil to function as a free radical scavenger and an antimicrobial food additive. RESULTS: When dissolved in ethanol or DMSO, the orange oil fraction had similar minimum inhibitory concentrations (MIC) for Listeria monocytogenes ATCC 19 115 (0.3% and 0.25% v/v respectively), which were significantly lower (P

Assuntos
Anti-Infecciosos/farmacologia , Citrus sinensis/química , Emulsificantes/química , Sequestradores de Radicais Livres/farmacologia , Óleos de Plantas/química , Óleos de Plantas/farmacologia , Anti-Infecciosos/química , Compostos de Bifenilo/química , Proliferação de Células/efeitos dos fármacos , Cicloexenos/análise , Dimetil Sulfóxido/química , Relação Dose-Resposta a Droga , Etanol/química , Aditivos Alimentares/química , Aditivos Alimentares/farmacologia , Manipulação de Alimentos , Sequestradores de Radicais Livres/química , Frutas/química , Radical Hidroxila/química , Lactobacillus plantarum/efeitos dos fármacos , Limoneno , Listeria monocytogenes/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Picratos/química , Polissorbatos/química , Salmonella typhimurium/efeitos dos fármacos , Solubilidade , Terpenos/análise , Fatores de Tempo
15.
Sensors (Basel) ; 10(4): 3562-84, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-22319312

RESUMO

Methionine is an essential amino acid for animals and is typically considered one of the first limiting amino acids in animal feed formulations. Methionine deficiency or excess in animal diets can lead to sub-optimal animal performance and increased environmental pollution, which necessitates its accurate quantification and proper dosage in animal rations. Animal bioassays are the current industry standard to quantify methionine bioavailability. However, animal-based assays are not only time consuming, but expensive and are becoming more scrutinized by governmental regulations. In addition, a variety of artifacts can hinder the variability and time efficacy of these assays. Microbiological assays, which are based on a microbial response to external supplementation of a particular nutrient such as methionine, appear to be attractive potential alternatives to the already established standards. They are rapid and inexpensive in vitro assays which are characterized with relatively accurate and consistent estimation of digestible methionine in feeds and feed ingredients. The current review discusses the potential to develop Escherichia coli-based microbial biosensors for methionine bioavailability quantification. Methionine biosynthesis and regulation pathways are overviewed in relation to genetic manipulation required for the generation of a respective methionine auxotroph that could be practical for a routine bioassay. A prospective utilization of Escherichia coli methionine biosensor would allow for inexpensive and rapid methionine quantification and ultimately enable timely assessment of nutritional profiles of feedstuffs.


Assuntos
Técnicas Biossensoriais/métodos , Escherichia coli/metabolismo , Metionina/análise , Metionina/farmacocinética , Bioensaio , Metionina/biossíntese , Mutação , Organismos Geneticamente Modificados
16.
Int J Food Microbiol ; 129(1): 43-9, 2009 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-19070381

RESUMO

Seven orange oil fractions were screened for their ability to inhibit the growth of selected Campylobacter and Arcobacter spp. using the standard agar-disk diffusion assay. Cold pressed (CP) terpeneless Valencia orange oil was found to be the most inhibitory to both Campylobacter jejuni and Campylobacter coli, exhibiting maximum zones of inhibition up to 80+/-0.0 mm. Five-fold concentrated Valencia oil and distilled d-limonene resulted in Campylobacter inhibition zones ranging from 11.0+/-1.4 to 44+/-1.4 mm against both C. jejuni and C. coli. No inhibition of Arcobacter spp. was detected by 6 out of 7 orange fractions except CP terpeneless Valencia orange oil which produced inhibition zones varying from 9.5+/-0.7 to 29+/-1.4 mm. Naturally occurring C. jejuni UAF 244 was isolated from a whole retail chicken, confirmed by hippuricase gene PCR assay, and used to determine antimicrobial capacities of the CP terpeneless Valencia orange oil and limonene when applied on chicken legs and thighs. The two types of chicken parts did not influence the antimicrobial strength of both orange fractions. While the observed reduction of C. jejuni cells attached to the skin varied approximately 1.5 to 2 logarithms compared to the control, the growth inhibition of the bacterial cells by limonene in the rinse increased by 6-fold and complete inhibition without recovery of detectable viable cells occurred when CP Valencia orange oil was applied. The study demonstrated the potential of the selected commercial orange oil fractions to serve as natural antimicrobials against C. jejuni, C. coli, and Arcobacter spp.


Assuntos
Antibacterianos/farmacologia , Arcobacter/efeitos dos fármacos , Campylobacter/efeitos dos fármacos , Galinhas/microbiologia , Cicloexenos/farmacologia , Óleos de Plantas/farmacologia , Terpenos/farmacologia , Animais , Arcobacter/crescimento & desenvolvimento , Campylobacter/crescimento & desenvolvimento , Contagem de Colônia Microbiana , Qualidade de Produtos para o Consumidor , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Relação Dose-Resposta a Droga , Microbiologia de Alimentos , Humanos , Limoneno , Testes de Sensibilidade Microbiana
17.
Sensors (Basel) ; 9(9): 7038-57, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-22399985

RESUMO

In animal diets optimal amino acid quantities and balance among amino acids is of great nutritional importance. Essential amino acid deficiencies have negative impacts on animal physiology, most often expressed in sub-optimal body weight gains. Over supplementation of diets with amino acids is costly and can increase the nitrogen emissions from animals. Although in vivo animal assays for quantification of amino acid bioavailability are well established, Escherichia coli-based bioassays are viable potential alternatives in terms of accuracy, cost, and time input. E. coli inhabits the gastrointestinal tract and although more abundant in colon, a relatively high titer of E. coli can also be isolated from the small intestine, where primary absorption of amino acids and peptides occur. After feed proteins are digested, liberated amino acids and small peptides are assimilated by both the small intestine and E. coli. The similar pattern of uptake is a necessary prerequisite to establish E. coli cells as accurate amino acid biosensors. In fact, amino acid transporters in both intestinal and E. coli cells are stereospecific, delivering only the respective biological l-forms. The presence of free amino- and carboxyl groups is critical for amino acid and dipeptide transport in both biological subjects. Di-, tri- and tetrapeptides can enter enterocytes; likewise only di-, tri- and tetrapeptides support E. coli growth. These similarities in addition to the well known bacterial genetics make E. coli an optimal bioassay microorganism for the assessment of nutritionally available amino acids in feeds.

18.
Foodborne Pathog Dis ; 5(5): 695-9, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18851678

RESUMO

Due to increasing concerns about the development of antimicrobial resistance amongst pathogenic bacteria, alternative strategies have been sought that do not use antibiotics to reduce pathogenic bacteria from foods and patients. A natural compound that has potent antimicrobial properties is citrus peel, which contains a variety of essential oils that inhibit the growth of or kill pathogenic bacteria. In the present study, seven citrus-based natural antimicrobials were evaluated for their ability to inhibit the growth of the pathogen Escherichia coli O157:H7. Zones of inhibition of E. coli O157:H7 by the citrus-derived fraction (10 microL/6 mm disk) were determined by a disk-diffusion assay on Sorbitol-MacConkey agar. Inhibition zones were observed after 48 h lawn growth of E. coli O157:H7 cells at 37 degrees C. Two citrus-based fractions, orange CP VAL terpeneless FAB 968611 and Limonene 1x Dist FAB 955430, inhibited E. coli O157:H7 with inhibition zones of approx. 11-24 mm dia. The remaining other five citrus-derived extracts (orange oil FL VAL 1121 ARR 974760, Orange 5x Conc VAL 4121 ARR 968374, orange terpenes ESS 1120 ARR 986259, orange terpenes CP 1100 ARR 986255, and orange terpenes OEO HP 1100 ARR 986257) were noninhibitory to E. coli O157:H7, yielding no clear inhibition zones. These studies show that citrus-derived natural compounds differ in their inhibitory activity against E. coli O157:H7 and some have potential applications as inhibitory agents against E. coli O157:H7 in various pathogen reduction strategies.


Assuntos
Antibacterianos/farmacologia , Citrus , Escherichia coli O157/efeitos dos fármacos , Microbiologia de Alimentos , Análise de Variância , Cicloexenos/farmacologia , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Escherichia coli O157/genética , Escherichia coli O157/crescimento & desenvolvimento , Limoneno , Óleos Voláteis/farmacologia , Terpenos/farmacologia
19.
Bioorg Khim ; 28(3): 269-76, 2002.
Artigo em Russo | MEDLINE | ID: mdl-12077854

RESUMO

The effect of 42 steroids of the 20-ketopregnane series with heterocycles fused in positions 16 alpha and 17 alpha on the activity of Na+,K(+)-activated ATPase from pig kidney was studied. It was shown that the studied compounds could be divided into two groups. The compounds from the first group stimulate the ATPase at low concentrations (1 x 10(-8)-1 x 10(-7) M) and inhibit it at high concentrations (1 x 10(-4) M). The second group of compounds stimulated the sodium pump at either concentration. This is explained by the cooperative action of the ATPase tetramer: after the reaction of its first binding site with the ligand, the tetramer changes the conformation and specificity of its other binding sites. Computer analysis of this series of compounds was carried out and a mathematical model of the dependence of their activities on the structure of their substituents was obtained with a high correlation coefficient and a satisfactory predictive power. This confirmed the structural similarity of the studied compounds with respect to their interaction with the ATPase binding sites. The method of descriptor analysis that was applied in this study is a new variant of approximation; it is based on the use of symbol variables as descriptors.


Assuntos
Mineralocorticoides/química , ATPase Trocadora de Sódio-Potássio/química , Algoritmos , Animais , Ativação Enzimática , Inibidores Enzimáticos/química , Ligantes , Modelos Biológicos , Ligação Proteica , Relação Estrutura-Atividade , Suínos
20.
Biofizika ; 23(3): 432-5, 1978.
Artigo em Russo | MEDLINE | ID: mdl-208653

RESUMO

The binding of spin label progesterone to bovine serum albumin was studied by the spin-probe technique. The binding capacity of protein was established. It was shown that protein formed a rigid complex with steroid, the correlation time of this complex being 50 ns. In the complex the radical part of the steroidal molecule has a hydrophobic environment.


Assuntos
Progesterona , Soroalbumina Bovina , Espectroscopia de Ressonância de Spin Eletrônica , Ligação Proteica , Marcadores de Spin
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