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1.
Fish Shellfish Immunol ; 144: 109244, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38000653

RESUMEN

Cell-mediated cytotoxicity is a complex immune mechanism that involves the release of several killing molecules, being perforin (PRF) one of the most important effector players. Perforin is synthesized by T lymphocytes and natural killer cells in mammals and responsible for the formation of pores on the target cell membrane during the killing process. Although perforin has been extensively studied in higher vertebrates, this knowledge is very limited in fish. Therefore, in this study we have identified four prf genes in European sea bass (Dicentrarchus labrax) and evaluated their mRNA levels. All sea bass prf genes showed the typical and conserved domains of its human orthologue and were closely clustered by the phylogenetic analysis. In addition, all genes showed constitutive and ubiquitous tissular expression, being prf1.9 gene the most highly expressed in immune tissues. Subsequently, in vitro stimulation of head-kidney (HK) cells with phytohemagglutinin, a T-cell activator, showed an increase of all prf gene levels, except for prf1.3 gene. European sea bass HK cells increased the transcription of prf1.2 and prf1.9 during the innate cell-mediated cytotoxic activity against xenogeneic target cells. In addition, sea bass infected with nodavirus (NNV) showed a similar expression pattern of all prf in HK and brain at 15 days post-infection, except for prf1.3 gene and in the gonad. Finally, the use of a polyclonal antibody against PRF1.9 showed an increase of positive cells in HK, brain and gonad from NNV-infected fish. Taken together, the data seem to indicate that all prf genes, except prf1.3, appear to be involved in the European sea bass immunity, and probably in the cell-mediated cytotoxic response, with PRF1.9 playing the most important role against nodavirus. The involvement of the PRFs and the CMC activity in the vertical transmission success of the virus is also discussed.


Asunto(s)
Lubina , Enfermedades de los Peces , Humanos , Animales , Filogenia , Perforina/genética , Mamíferos
2.
J Dairy Sci ; 107(7): 4277-4287, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38395395

RESUMEN

Cheese is a food in which toxic concentrations of biogenic amines (BA) may be reached, mainly as a consequence of the decarboxylation of determined amino acids by certain lactic acid bacteria (LAB). To maintain the food safety of cheese, environmentally friendly strategies are needed that specifically prevent the growth of BA-producing LAB and the accumulation of BA. The bacteriocins produced by LAB are natural compounds with great potential as food biopreservatives. This work examines the antimicrobial potential of 7 bacteriocin-containing, cell-free supernatants (CFS: coagulin A-CFS, enterocin A-CFS, enterocin P-CFS, lacticin 481-CFS, nisin A-CFS, nisin Z-CFS and plantaricin A-CFS) produced by LAB against 48 strains of the LAB species largely responsible for the accumulation of the most important BA in cheese, that is, histamine, tyramine, and putrescine. Susceptibility to the different CFS was strain-dependent. The histamine-producing species with the broadest sensitivity spectrum were Lentilactobacillus parabuchneri (the species mainly responsible for the accumulation of histamine in cheese) and Pediococcus parvulus. The tyramine-producing species with the broadest sensitivity spectrum was Enterococcus faecium, and Enterococcus faecalis and Enterococcus hirae were among the most sensitive putrescine producers. Nisin A-CFS was active against 31 of the 48 BA-producing strains (the broadest antimicrobial spectrum recorded). Moreover, commercial nisin A prevented biofilm formation by 67% of the BA-producing, biofilm-forming LAB strains. These findings underscore the potential of bacteriocins in the control of BA-producing LAB and support the use of nisin A as a food-grade biopreservative for keeping BA-producing LAB in check and reducing BA accumulation in cheese.


Asunto(s)
Bacteriocinas , Biopelículas , Aminas Biogénicas , Queso , Lactobacillales , Nisina , Queso/microbiología , Bacteriocinas/farmacología , Bacteriocinas/metabolismo , Aminas Biogénicas/metabolismo , Nisina/farmacología , Biopelículas/efectos de los fármacos , Lactobacillales/metabolismo , Antiinfecciosos/farmacología , Microbiología de Alimentos
3.
Arch Microbiol ; 204(4): 220, 2022 Mar 25.
Artículo en Inglés | MEDLINE | ID: mdl-35333989

RESUMEN

Currently, consumption of spontaneously fermented milks is common in Algeria, making it a feasible source of diverse lactic acid bacteria (LAB) with the potential to be used as adjunct cultures to improve quality and safety of fermented dairy products. In this context, to select eligible indigenous strains which could be applied as bioprotective and/or starter cultures, the present study aimed to characterize the genomic variability, biotechnological potential, and safety of thirty-eight LAB isolated from Algerian dairy and farm sources of western Algeria. The isolates were unequivocally identified by 16S rRNA gene and fingerprint-based methods. The following species were identified: Enterococcus faecium (n = 15), Enterococcus durans (n = 2), Enterococcus hirae (n = 2), Enterococcus lactis (n = 1), Lactiplantibacillus plantarum (n = 6), Lactococcus lactis (n = 4), Levilactobacillus brevis (n = 3), Lacticaseibacillus paracasei (n = 3), Lacticaseibacillus rhamnosus (n = 1), and Pediococcus acidilactici (n = 1). Among the strains, three of them, L. lactis LGMY8, Lb. plantarum LGMY30 and Lb. paracasei LGMY31 were safe and showed some valuable biotechnological properties, such as high acidification, proteolytic activity, EPS production, and inhibition of undesirable bacteria that made them powerful candidates to be used as starter.


Asunto(s)
Lactobacillales , Argelia , Granjas , Microbiología de Alimentos , ARN Ribosómico 16S/genética
4.
Environ Res ; 200: 111750, 2021 09.
Artículo en Inglés | MEDLINE | ID: mdl-34303683

RESUMEN

The objective of this study was to analyze the influence of different operational variables (catalyst loading, initial EtP concentration, medium pH, the presence of anions and radical scavengers) on the performance of ethylparaben (EtP) photodegradation catalyzed with an rGO/TiO2 composite. EtP was selected for study after analyzing the effect of paraben chain length on its catalytic photodegradation, finding that the photodegradation rate constant values of methyl-, ethyl-, and butyl-paraben are 0.050, 0.096, and 0.136 min-1, respectively. This indicates that the photodegradation rate constant of parabens is higher with longer alkyl chain, which augments its oxidation capacity. The percentage removal of EtP at 40 min increases from 66.3 to 98.6 % when the composite dose rises from 100 to 700 mg/L; however, an additional increase in the composite dose to 1000 mg/L does not substantively improve the photodegradation rate or percentage EtP removal (98.9 %). A rise in the initial EtP concentration from 15 to 100 mg/L reduces the percentage of degradation from 100 to 76.4 %. The percentage EtP degradation is lower with higher solution pH. The presence of HCO3- or Cl- anions in the medium reduces the degradation performance. Results obtained using positive hole and hydroxyl radical scavengers demonstrate that positive holes play an important role in EtP degradation. No degradation product evidences toxicity against the cultured human embryonic kidney cell line HEK-293.


Asunto(s)
Parabenos , Contaminantes Químicos del Agua , Grafito , Células HEK293 , Humanos , Fotólisis , Titanio , Rayos Ultravioleta , Contaminantes Químicos del Agua/análisis
5.
J Environ Manage ; 281: 111871, 2021 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-33385896

RESUMEN

Manganese ferrite solid nanospheres (MSNs) were prepared by a solvothermal method and calcined at various temperatures up to 500 °C. Their surface area, morphology, particle size, weight change during calcination, surface coordination number of metal ions, oxidation state, crystal structure, crystallite size, and magnetic properties were studied. The MSNs were used as catalysts to activate potassium peroxymonosulfate (PMS) for the oxidative degradation of para-nitrophenol (PNP) from water and for the oxidation of n-C7 asphaltenes in flowing air at atmospheric (0.084 MPa) and high pressure (6 MPa). Mn was in oxidation states (II) and (III) at calcination temperature of 200 °C, and the crystalline structure corresponded to jacobsite. Mn was in oxidation states (III) and (IV) at 350 °C and in oxidation states (II), (III), and (IV) at 500 °C, and the crystalline structure was maghemite at both temperatures. MSN catalysts generated hydroxyl (HO·) and sulfate (SO4·-) radicals in the PMS activation and generated HO· radicals in the n-C7 asphaltene oxidation. In both reactions, the best catalyst was MSN calcined at 350 °C (MSN350), because it has the highest concentration of Mn(III) in octahedral B sites, which are directly exposed to the catalyst surface, and the largest total and lattice oxygen contents, favoring oxygen mobility for Mn redox cycles. The MSN350 sample reduces the decomposition temperature of n-C7 asphaltenes from 430 to 210 °C at 0.084 MPa and from 370 to 200 °C at 6.0 MPa. In addition, it reduces the effective activation energy by approximately 77.6% in the second combustion (SC) region, where high-temperature oxidation reactions take place.


Asunto(s)
Nanosferas , Catálisis , Nitrofenoles , Oxidación-Reducción , Peróxidos , Hidrocarburos Policíclicos Aromáticos
6.
BMC Nephrol ; 21(1): 227, 2020 06 15.
Artículo en Inglés | MEDLINE | ID: mdl-32539688

RESUMEN

BACKGROUND: Acute kidney injury (AKI) occurs in 12-20% of multiple myeloma (MM) patients. Several studies have shown a reduction of free light chains (FLC) using hemodialysis with High-Cut-Off membranes. However, this technique entails albumin loss. Hemodiafiltration with ultrafiltrate regeneration is a technique that includes a process of adsorption. The aim of this study was to evaluate the effectiveness of hemodiafiltration with ultrafiltrate regeneration in reducing FLC levels without causing albumin loss. METHODS: This is an observational study (2012 to 2018) including nine patients with MM (5 kappa, 4 lambda) and AKI. All patients were treated with chemotherapy and hemodiafiltration with ultrafiltrate regeneration. Blood Samples (pre and post-dialysis) and ultrafiltrate were collected pre and post-resin at 5 min after initiation of the session and 5 min before the end of the procedure. RESULTS: The serum levels of kappa and lambda were reduced by a 57.6 ± 10% and 33.5 ± 25% respectively. Serum albumin concentration remained unchanged after the procedure. In the ultrafiltrate, the mean FLC reduction ratio shortly after initiation of the dialysis procedure was: 99.2 and 97.06% for kappa and lambda respectively, and only 0.7% for albumin; and at the end of the session the percent reduction was: 63.7 and 33.62% for kappa and lambda respectively, and 0.015% for albumin. Patients clinical outcome was: 33.3% recovered renal function, 22.2% died during the first year and 44.4% required maintenance dialysis. CONCLUSIONS: Hemodiafiltration with ultrafiltrate regeneration reduces FLC levels without producing a significant loss of albumin; and, FLC removal is maintained throughout the session. Therefore, hemodiafiltration with ultrafiltrate regeneration may be considered an effective adjunctive therapy in patients with MM.


Asunto(s)
Lesión Renal Aguda/sangre , Hemodiafiltración/métodos , Cadenas kappa de Inmunoglobulina/sangre , Cadenas lambda de Inmunoglobulina/sangre , Mieloma Múltiple/sangre , Albúmina Sérica/análisis , Lesión Renal Aguda/etiología , Lesión Renal Aguda/terapia , Anciano , Anciano de 80 o más Años , Creatinina/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Mieloma Múltiple/complicaciones
7.
Molecules ; 25(24)2020 Dec 21.
Artículo en Inglés | MEDLINE | ID: mdl-33371436

RESUMEN

In the present study, the synthesis of gold nanoparticles (AuNPs) loaded with methotrexate (MTX) has been carried out in order to obtain controlled size and monodispersed nanocarriers of around 20 nm. The characterization study shows metallic AuNPs with MTX polydispersed on the surface. MTX is linked by the replacement of citrate by the MTX carboxyl group. The drug release profiles show faster MTX release when it is conjugated, which leads to the best control of plasma concentration. Moreover, the enhanced release observed at pH 5 could take advantage of the pH gradients that exist in tumor microenvironments to achieve high local drug concentrations. AuNP-MTX conjugates were tested by flow cytometry against lung (A-549) and colon (HTC-116) cancer cell lines. Results for A-549 showed a weaker dose-response effect than for colon cancer ones. This could be related to the presence of folate receptors in line HTC-116 in comparison to line A-549, supporting the specific uptake of folate-conjugated AuNP-MTX by folate receptor positive tumor cells. Conjugates exhibited considerably higher cytotoxic effects compared with the effects of equal doses of free MTX. Annexin V-PI tests sustained the cell death mechanism of apoptosis, which is normally disabled in cancer cells.


Asunto(s)
Antineoplásicos/farmacología , Neoplasias del Colon/tratamiento farmacológico , Oro/química , Neoplasias Pulmonares/tratamiento farmacológico , Nanopartículas del Metal/química , Metotrexato/farmacología , Células A549 , Anexina A5/metabolismo , Antineoplásicos/química , Apoptosis/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Línea Celular Tumoral , Colon/efectos de los fármacos , Colon/metabolismo , Neoplasias del Colon/metabolismo , Sistemas de Liberación de Medicamentos/métodos , Liberación de Fármacos/fisiología , Ácido Fólico/farmacología , Células HCT116 , Humanos , Concentración de Iones de Hidrógeno , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Neoplasias Pulmonares/metabolismo , Metotrexato/química , Microambiente Tumoral/efectos de los fármacos
8.
Food Technol Biotechnol ; 58(3): 260-272, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-33281482

RESUMEN

RESEARCH BACKGROUND: Consumption of spontaneously fermented camel´s milk is common in Algeria, making it a feasible source of diverse lactic acid bacteria (LAB) with the potential to be used as adjunct cultures to improve quality and safety of fermented dairy products. EXPERIMENTAL APPROACH: Twelve raw camel´s milk samples were used as a source of indigenous LAB, which were further characterised by examining39 phenotypic traits with technological relevance. RESULTS AND CONCLUSIONS: Thirty-five non-starter LAB (NSLAB) were isolated from 12 Algerian raw camel's milk samples and they were microbiologically, biochemically and genetically characterised. Some isolates showed proteolytic activity, acidifying capacity, the ability to use citrate, and to produce dextran and acetoin. Ethanol, acetaldehyde, methyl acetate, acetoin and acetic acid were the major volatile compounds detected. Cluster analysis performed using the unweighted group with arithmetic average (UPGMA) method, and based on the thirty-nine phenotypic characteristics investigated, reflected the microbial diversity that can be found in raw camel´s milk. NOVELTY AND SCIENTIFIC CONTRIBUTION: The isolated strains, from a non-typical source, showed interesting technological traits to be considered as potential adjunct cultures. Cluster analysis based on the examined phenotypic characteristics proved to be a useful tool for the typification of isolates when no genetic information is available. These findings may be of use towards an industrialised production of camel's milk dairy products.

9.
Anal Bioanal Chem ; 411(25): 6615-6624, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31359119

RESUMEN

Histamine, a biogenic amine, is abundant in fermented foods and beverages, notably wine. A high intake of this monoamine may produce adverse reactions in humans, which may be severe in individuals with a reduced capacity to catabolise extrinsic histamine. Thus, control of histamine concentration during wine production and before distribution is advisable. Simple, rapid, point-of-use bioanalytical platforms are needed because traditional methods for the detection and quantification of histamine are expensive and time-consuming. This work applies the lateral flow immunoassay technique to histamine detection. Superparamagnetic particle labels, and an inductive sensor designed to read the test line in the immunoassay, enable magnetic quantification of the molecule. The system is calibrated with histamine standards in the interval of interest for wine production. A commercial optical strip reader is used for comparison measurements. The lateral flow system has a limit of detection of 1.2 and 1.5 mg/L for the inductive and optical readers, respectively. The capability of the inductive system for histamine quantification is demonstrated for wine samples at different processing points (at the end of alcoholic fermentation, at the end of malolactic fermentation, in freshly bottled wine, and in reserve wine). The results are validated by ultra-high-performance liquid chromatography. Graphical abstract.


Asunto(s)
Histamina/análisis , Vino/análisis , Aminas Biogénicas/análisis , Diseño de Equipo , Inmunoensayo/métodos , Límite de Detección , Nanopartículas de Magnetita/química , Tiras Reactivas/análisis
10.
Fam Pract ; 36(6): 693-698, 2019 11 18.
Artículo en Inglés | MEDLINE | ID: mdl-31044230

RESUMEN

BACKGROUND: Although both hospitalization and mortality due to heart failure (HF) have been widely studied, less is known about the impact of HF on disability and quality of life. AIM: To assess the degree of disability and quality of life in HF patients attended at family medicine centres. DESIGN AND SETTING: Cross-sectional study of a cohort of HF patients attended at family medicine centres. METHODS: Disability was assessed with the WHODAS 2 questionnaire, which provides a global and six domain scores that is understanding and communication, getting around, self-care, getting along with people, life activities and participation in society. Quality of life was assessed with the Minnesota Living with Heart Failure Questionnaire, which furnishes a global and two domain scores, physical and emotional. RESULTS: A breakdown of the results showed that 28% of patients had moderate disability and 16.7% had severe disability, with the most important areas affected being: life activities, 8.9% extreme disability and 30.3% severe disability; getting around, 34.6% severe disability and 2% extreme disability; and participation in society, 53.3% moderate-severe disability. Quality of life was mildly affected. New York Heart Association (NYHA) Functional Classification and sex were the major determinants of disability and quality of life. Angiotensin-converting enzyme inhibitors and angiotensin II receptor antagonists were associated with better scores in the "getting around" and "life activity" domains. CONCLUSION: HF patients in primary care show an important degree of disability and an acceptable quality of life.


Asunto(s)
Evaluación de la Discapacidad , Personas con Discapacidad/estadística & datos numéricos , Insuficiencia Cardíaca/tratamiento farmacológico , Insuficiencia Cardíaca/fisiopatología , Calidad de Vida , Actividades Cotidianas , Anciano , Anciano de 80 o más Años , Antagonistas de Receptores de Angiotensina/uso terapéutico , Inhibidores de la Enzima Convertidora de Angiotensina/uso terapéutico , Estudios Transversales , Femenino , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Análisis Multivariante , Atención Primaria de Salud , Autocuidado/estadística & datos numéricos , España , Encuestas y Cuestionarios
11.
Can J Microbiol ; 62(5): 422-30, 2016 May.
Artículo en Inglés | MEDLINE | ID: mdl-27021684

RESUMEN

A selective culture medium containing acid-hydrolyzed gliadins as the sole nitrogen source was used in the search for sourdough-indigenous lactic acid bacteria (LAB) with gliadin-metabolizing activity. Twenty gliadin-degrading LAB strains were isolated from 10 sourdoughs made in different ways and from different geographical regions. Fifteen of the 20 isolated strains were identified as Lactobacillus casei, a species usually reported as subdominant in sourdough populations. The other 5 gliadin-degrading strains belonged to the more commonly encountered sourdough species Leuconostoc mesenteroides and Lactobacillus plantarum. All these strains were shown to be safe in terms of their resistance to antimicrobial agents. When individually incubated with the α2-gliadin-derived immunotoxic 33-mer peptide (97.5 ppm), half of the L. casei strains metabolized at least 50% of it within 24 h. One strain metabolized 82% of the 33-mer peptide within 8 h and made it fully disappear within 12 h. These results reveal for the first time the presence in sourdough of proteolytic L. casei strains with the capacity to individually metabolize the coeliac-disease-related 33-mer peptide.


Asunto(s)
Pan/microbiología , Gliadina/metabolismo , Lacticaseibacillus casei/metabolismo , Fragmentos de Péptidos/metabolismo , Fermentación , Hidrólisis , Lacticaseibacillus casei/aislamiento & purificación , Lactobacillus plantarum/metabolismo , Péptidos/metabolismo
12.
Food Microbiol ; 59: 85-91, 2016 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-27375247

RESUMEN

The consumption of food containing large amounts of histamine can lead to histamine poisoning. Cheese is one of the most frequently involved foods. Histamine, one of the biogenic amines (BAs) exhibiting the highest safety risk, accumulates in food contaminated by microorganisms with histidine decarboxylase activity. The origin of these microorganisms may be very diverse with contamination likely occurring during post-ripening processing, but the microorganisms involved during this manufacturing step have never been identified. The present work reports the isolation of 21 histamine-producing Lactobacillus parabuchneri strains from a histamine-containing grated cheese. PCR revealed that every isolate carried the histidine decarboxylase gene (hdcA). Eight lineages were identified based on the results of genome PFGE restriction analysis plus endonuclease restriction profile analysis of the carried plasmids. Members of all lineages were able to form biofilms on polystyrene and stainless steel surfaces. L. parabuchneri is therefore an undesirable species in the dairy industry; the biofilms it can produce on food processing equipment represent a reservoir of histamine-producing bacteria and thus a source of contamination of post-ripening-processed cheeses.


Asunto(s)
Biopelículas/crecimiento & desarrollo , Queso/microbiología , Microbiología de Alimentos , Histamina/biosíntesis , Lactobacillus/aislamiento & purificación , Lactobacillus/fisiología , Acero Inoxidable , Queso/análisis , Electroforesis en Gel de Campo Pulsado , Contaminación de Alimentos/prevención & control , Manipulación de Alimentos , Histidina Descarboxilasa/genética , Lactobacillus/genética , Lactobacillus/metabolismo , Reacción en Cadena de la Polimerasa , Poliestirenos
13.
Appl Environ Microbiol ; 81(18): 6145-57, 2015 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-26116671

RESUMEN

Dairy industry fermentative processes mostly use Lactococcus lactis as a starter. However, some dairy L. lactis strains produce putrescine, a biogenic amine that raises food safety and spoilage concerns, via the agmatine deiminase (AGDI) pathway. The enzymatic activities responsible for putrescine biosynthesis in this bacterium are encoded by the AGDI gene cluster. The role of the catabolic genes aguB, aguD, aguA, and aguC has been studied, but knowledge regarding the role of aguR (the first gene in the cluster) remains limited. In the present work, aguR was found to be a very low level constitutively expressed gene that is essential for putrescine biosynthesis and is transcribed independently of the polycistronic mRNA encoding the catabolic genes (aguBDAC). In response to agmatine, AguR acts as a transcriptional activator of the aguB promoter (PaguB), which drives the transcription of the aguBDAC operon. Inverted sequences required for PaguB activity were identified by deletion analysis. Further work indicated that AguR is a transmembrane protein which might function as a one-component signal transduction system that senses the agmatine concentration of the medium and, accordingly, regulates the transcription of the aguBDAC operon through a C-terminal cytoplasmic DNA-binding domain typically found in LuxR-like proteins.


Asunto(s)
Agmatina/metabolismo , Lactococcus lactis/genética , Lactococcus lactis/metabolismo , Transactivadores/genética , Transactivadores/metabolismo , Transcripción Genética , Vías Biosintéticas/genética , Perfilación de la Expresión Génica , Regulación Bacteriana de la Expresión Génica , Operón , Putrescina/biosíntesis
14.
Appl Environ Microbiol ; 81(17): 5784-93, 2015 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-26092449

RESUMEN

Rotavirus is the leading cause of infantile diarrhea in developing countries, where it causes a high number of deaths among infants. Two vaccines are available, being highly effective in developed countries although markedly less efficient in developing countries. As a complementary treatment to the vaccines, a Lactobacillus strain producing an anti-rotavirus antibody fragment in the gastrointestinal tract could potentially be used. In order to develop such an alternative therapy, the effectiveness of Lactobacillus rhamnosus GG to produce and display a VHH antibody fragment (referred to as anti-rotavirus protein 1 [ARP1]) on the surface was investigated. L. rhamnosus GG is one of the best-characterized probiotic bacteria and has intrinsic antirotavirus activity. Among four L. rhamnosus GG strains [GG (CMC), GG (ATCC 53103), GG (NCC 3003), and GG (UT)] originating from different sources, only GG (UT) was able to display ARP1 on the bacterial surface. The genomic analysis of strain GG (UT) showed that the genes welE and welF of the EPS cluster are inactivated, which causes a defect in exopolysaccharide (EPS) production, allowing efficient display of ARP1 on its surface. Finally, GG (UT) seemed to confer a level of protection against rotavirus-induced diarrhea similar to that of wild-type GG (NCC 3003) in a mouse pup model, indicating that the EPS may not be involved in the intrinsic antirotavirus activity. Most important, GG (EM233), a derivative of GG (UT) producing ARP1, was significantly more protective than the control strain L. casei BL23.


Asunto(s)
Proteínas Bacterianas/metabolismo , Fragmentos de Inmunoglobulinas/metabolismo , Lacticaseibacillus rhamnosus/metabolismo , Polisacáridos Bacterianos/deficiencia , Infecciones por Rotavirus/microbiología , Rotavirus/fisiología , Animales , Proteínas Bacterianas/genética , Modelos Animales de Enfermedad , Femenino , Humanos , Fragmentos de Inmunoglobulinas/genética , Lacticaseibacillus rhamnosus/genética , Masculino , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Probióticos/administración & dosificación , Infecciones por Rotavirus/prevención & control , Infecciones por Rotavirus/virología
15.
Microb Cell Fact ; 14: 208, 2015 Dec 30.
Artículo en Inglés | MEDLINE | ID: mdl-26715338

RESUMEN

BACKGROUND: Lactococcus lactis has been safely consumed in fermented foods for millennia. This Gram-positive bacterium has now become of industrial importance as an expression host for the overproduction of lipopolysaccharide-free recombinant proteins used as food ingredients, therapeutic proteins and biotechnological enzymes. RESULTS: This paper reports an agmatine-controlled expression (ACE) system for L. lactis, comprising the lactococcal agmatine-sensor/transcriptional activator AguR and its target promoter P(aguB). The usefulness and efficiency of this system was checked via the reporter gene gfp and by producing PEP (Myxococcus xanthus prolyl-endopeptidase), an enzyme of biomedical interest able to degrade the immunotoxic peptides produced during the gastrointestinal breakdown of gluten. CONCLUSION: The ACE system developed in this work was suitable for the efficient expression of the functional recombinant proteins GFP and PEP. The expression system was tightly regulated by the agmatine concentration and allowed high protein production without leakiness.


Asunto(s)
Agmatina/metabolismo , Regulación Bacteriana de la Expresión Génica/genética , Lactococcus lactis/genética , Lactococcus lactis/metabolismo
16.
Appl Microbiol Biotechnol ; 99(2): 897-905, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25341400

RESUMEN

Lactococcus lactis is the most important starter culture organism used in the dairy industry. Although L. lactis species have been awarded Qualified Presumption of Safety status by the European Food Safety Authority, and Generally Regarded as Safe status by the US Food and Drug Administration, some strains can produce the biogenic amine putrescine. One such strain is L. lactis subsp. cremoris CECT 8666 (formerly L. lactis subsp. cremoris GE2-14), which was isolated from Genestoso cheese. This strain catabolizes agmatine to putrescine via the agmatine deiminase (AGDI) pathway, which involves the production of ATP and two ammonium ions. The present work shows that the availability of agmatine and its metabolization to putrescine allows for greater bacterial growth (in a biphasic pattern) and causes the alkalinization of the culture medium in a dose-dependent manner. The construction of a mutant lacking the AGDI cluster (L. lactis CECT 8666 Δagdi) confirmed the latter's direct role in putrescine production, growth, and medium alkalinization. Alkalinization did not affect the putrescine production pattern and was not essential for increased bacterial growth.


Asunto(s)
Proteínas Bacterianas/metabolismo , Medios de Cultivo/química , Hidrolasas/metabolismo , Lactococcus lactis/crecimiento & desarrollo , Putrescina/biosíntesis , Compuestos de Amonio/metabolismo , Proteínas Bacterianas/genética , Queso/análisis , ADN Bacteriano/genética , Fermentación , Inocuidad de los Alimentos , Concentración de Iones de Hidrógeno , Hidrolasas/genética , Lactococcus lactis/genética , Familia de Multigenes , Mutación
17.
Appl Microbiol Biotechnol ; 99(8): 3547-58, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25529314

RESUMEN

Enterococcus faecalis is a commensal bacterium of the human gut that requires the ability to pass through the stomach and therefore cope with low pH. E. faecalis has also been identified as one of the major tyramine producers in fermented food products, where they also encounter acidic environments. In the present work, we have constructed a non-tyramine-producing mutant to study the role of the tyramine biosynthetic pathway, which converts tyrosine to tyramine via amino acid decarboxylation. Wild-type strain showed higher survival in a system that mimics gastrointestinal stress, indicating that the tyramine biosynthetic pathway has a role in acid resistance. Transcriptional analyses of the E. faecalis V583 tyrosine decarboxylase cluster showed that an acidic pH, together with substrate availability, induces its expression and therefore the production of tyramine. The protective role of the tyramine pathway under acidic conditions appears to be exerted through the maintenance of the cytosolic pH. Tyramine production should be considered important in the adaptability of E. faecalis to acidic environments, such as fermented dairy foods, and to survive passage through the human gastrointestinal tract.


Asunto(s)
Enterococcus faecalis/genética , Enterococcus faecalis/metabolismo , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Transcripción Genética/efectos de los fármacos , Tiramina/biosíntesis , Perfilación de la Expresión Génica , Técnicas de Inactivación de Genes , Humanos , Concentración de Iones de Hidrógeno , Viabilidad Microbiana/efectos de los fármacos , Familia de Multigenes , Tirosina Descarboxilasa/biosíntesis , Tirosina Descarboxilasa/genética
18.
Can J Microbiol ; 61(7): 517-9, 2015 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-25966618

RESUMEN

Enterococcus faecalis is one of the most controversial species of lactic acid bacteria. Some strains are used as probiotics, while others are associated with severe and life-threatening nosocomial infections. Their pathogenicity depends on the acquisition of multidrug resistance and virulence factors. Gelatinase, which is required in the first steps of biofilm formation, is an important virulence determinant involved in E. faecalis pathogenesis, including endocarditis and peritonitis. The gene that codes for gelatinase (gelE) is controlled by the Fsr quorum-sensing system, whose encoding genes (fsrA, fsrB, fsrC, and fsrD) are located immediately upstream of gelE. The integration of a DNA fragment into the fsr locus of a derived mutant of E. faecalis V583 suppressed the gelatinase activity and prevented biofilm formation. Sequence analysis indicated the presence of IS256 integrated into the fsrC gene at nucleotide position 321. Interestingly, IS256 is also associated with biofilm formation in Staphylococcus epidermidis and Staphylococcus aureus. This is the first description of an insertion sequence that prevents biofilm formation in E. faecalis.


Asunto(s)
Proteínas Bacterianas/metabolismo , Biopelículas , Infección Hospitalaria/microbiología , Elementos Transponibles de ADN , Enterococcus faecalis/enzimología , Enterococcus faecalis/fisiología , Gelatinasas/metabolismo , Infecciones por Bacterias Grampositivas/microbiología , Proteínas Bacterianas/genética , Enterococcus faecalis/genética , Gelatinasas/genética , Humanos , Mutagénesis Insercional , Percepción de Quorum
19.
Food Microbiol ; 48: 163-70, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25791004

RESUMEN

Lactococcus lactis is the lactic acid bacterial (LAB) species most widely used as a primary starter in the dairy industry. However, several strains of L. lactis produce the biogenic amine putrescine via the agmatine deiminase (AGDI) pathway. We previously reported the putrescine biosynthesis pathway in L. lactis subsp. cremoris GE2-14 to be regulated by carbon catabolic repression (CCR) via glucose but not lactose (Linares et al., 2013). The present study shows that both these sugars repress putrescine biosynthesis in L. lactis subsp. lactis T3/33, a strain isolated from a Spanish artisanal cheese. Furthermore, we demonstrated that both glucose and lactose repressed the transcriptional activity of the aguBDAC catabolic genes of the AGDI route. Finally, a screening performed in putrescine-producing dairy L. lactis strains determined that putrescine biosynthesis was repressed by lactose in all the L. lactis subsp. lactis strains tested, but in only one L. lactis subsp. cremoris strain. Given the obvious importance of the lactose-repression in cheese putrescine accumulation, it is advisable to consider the diversity of L. lactis in this sense and characterize consequently the starter cultures to select the safest strains.


Asunto(s)
Represión Catabólica , Queso/microbiología , Lactococcus lactis/metabolismo , Lactosa/metabolismo , Putrescina/biosíntesis , Animales , Bovinos , Glucosa/metabolismo , Leche/microbiología
20.
Water Sci Technol ; 71(5): 789-94, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25768228

RESUMEN

The objective of this study was to investigate the effect of the presence of an activated carbon cloth (ACC) during the degradation and removal of gallic acid (GA) and p-coumaric acid (pCA) by Fenton oxidation using H2O2 and FeSO4 as catalyst. Removal of GA or pCA by Fenton oxidation was much higher than that of total organic carbon (TOC), indicating that a large proportion of GA or pCA degradation products was not mineralized. The presence of ACC increased the concentration of hydroxyl radicals generated in the FeSO4 + H2O2 system. The presence of ACC during Fenton oxidation largely increased TOC and GA removal, attributable to the adsorption of GA and its degradation products and the increased generation of OH(•) radicals that mineralize them. In the Fenton oxidation of pCA, the presence of ACC produced the same effects as for GA, but now the increased removal of pCA was due to adsorption on the activated carbon and not to the increased generation of hydroxyl radicals, due to the greater affinity of pCA for the carbon surface and its more difficult mineralization in comparison to GA.


Asunto(s)
Carbón Orgánico , Ácidos Cumáricos/química , Ácido Gálico/química , Adsorción , Carbono , Catálisis , Compuestos Férricos/química , Peróxido de Hidrógeno/química , Radical Hidroxilo , Oxidación-Reducción , Propionatos , Agua
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