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1.
PLoS One ; 19(1): e0294494, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38170702

RESUMEN

Establishing the infant's gut microbiota has long-term implications on health and immunity. Breastfeeding is recognized as the best practice of infant nutrition in comparison with formula feeding. We evaluated the effects of the primary feeding practices by analyzing the infant growth and the potential association with gut diseases. A cross-sectional and observational study was designed. This study included 55 mothers with infants, who were divided according to their feeding practices in breastfeeding (BF), formula feeding (FF), and combined breast and formula feeding (CF). Anthropometric measurements of the participants were recorded. Additionally, non-invasive fecal samples from the infants were collected to analyze the microbiota by sequencing, immunoglobulin A (IgA) concentration (ELISA), and volatile organic compounds (gas chromatography with an electronic nose). Results showed that the microbiota diversity in the BF group was the highest compared to the other two groups. The IgA levels in the BF group were twice as high as those in the FF group. Moreover, the child´s growth in the BF group showed the best infant development when the data were compared at birth to the recollection time, as noted by the correlation with a decreased concentration of toxic volatile organic compounds. Interestingly, the CF group showed a significant difference in health status when the data were compared with the FF group. We conclude that early health practices influence children's growth, which is relevant to further research about how those infants' health evolved.


Asunto(s)
Microbioma Gastrointestinal , Compuestos Orgánicos Volátiles , Recién Nacido , Lactante , Femenino , Niño , Humanos , Estudios Transversales , Lactancia Materna , Inmunoglobulina A , Fórmulas Infantiles
2.
PLoS One ; 18(5): e0286079, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37224160

RESUMEN

CONTEXT: Periodontitis is a chronic multifactorial inflammatory disease linked to oral microbiota dysbiosis. This disease progresses to infection that stimulates a host immune/inflammatory response, with progressive destruction of the tooth-supporting structures. OBJECTIVE: This systematic review aims to present a robust critical evaluation of the evidence of salivary protein profiles for identifying oral diseases using proteomic approaches and summarize the use of these approaches to diagnose chronic periodontitis. DATA SOURCES: A systematic literature search was conducted from January 1st, 2010, to December 1st, 2022, based on PICO criteria following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines and by searching the three databases Science Direct, Scopus, and Springer Link. STUDY SELECTION: According to the inclusion criteria, eight studies were identified to analyze the proteins identified by proteomics. RESULTS: The protein family S100 was identified as the most abundant in patients with chronic periodontitis. In this family, an increased abundance of S100A8 and S100A9 from individuals with the active disease was observed, which strongly relates to the inflammatory response. Moreover, the ratio S100A8/S100A9 and the metalloproteinase-8 in saliva could differentiate distinct periodontitis groups. The changes in protein profile after non-surgical periodontal therapy improved the health of the buccal area. The results of this systematic review identified a set of proteins that could be used as a complementary tool for periodontitis diagnosis using salivary proteins. CONCLUSION: Biomarkers in saliva can be used to monitor an early stage of periodontitis and the progression of the disease following therapy.


Asunto(s)
Periodontitis Crónica , Humanos , Periodontitis Crónica/diagnóstico , Periodontitis Crónica/terapia , Proteómica , Saliva , Periodoncio , Ligamento Periodontal , Calgranulina A , Calgranulina B
3.
Plants (Basel) ; 12(3)2023 Jan 21.
Artículo en Inglés | MEDLINE | ID: mdl-36771585

RESUMEN

Plant-associated bacteria in heavy-metal-contaminated environments could be a biotechnological tool to improve plant growth. The present work aimed to isolate lead- and cadmium-tolerant endophytic bacteria from the roots of Typha latifolia growing in a site contaminated with these heavy metals. Endophytic bacteria were characterized according to Pb and Cd tolerance, plant-growth-promoting rhizobacteria activities, and their effect on T. latifolia seedlings exposed and non-exposed to Pb and Cd. Pb-tolerant isolates were identified as Pseudomonas azotoformans JEP3, P. fluorescens JEP8, and P. gessardii JEP33, while Cd-tolerant bacteria were identified as P. veronii JEC8, JEC9, and JEC11. They all exert biochemical activities, including indole acetic acid synthesis, siderophore production, and phosphate solubilization. Plant-bacteria interaction assays showed that P. azotoformans JEP3, P. fluorescens JEP8, P. gessardii JEP33, and P. veronii JEC8, JEC9, JEC11 promote the growth of T. latifolia seedlings by increasing the root and shoot length, while in plants exposed to either 5 mg/L of Pb or 10 mg/L of Cd, all bacterial isolates increased the shoot length and the number of roots per plant, suggesting that they are plant-growth-promoting rhizobacteria that could contribute to T. latifolia adaptation to the heavy metal polluted site.

4.
3 Biotech ; 12(9): 210, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35945985

RESUMEN

In this work, we evaluated the removal efficiency of diclofenac by Chlorella vulgaris OW-01, Nannochloropsis oculata CCAP 849/7, Scenedesmus acutus UTEX 72, and Scenedesmus obliquus CCAP 276/2. Each microalga was grown in media with different concentrations (50 and 100% of the original formulation) of carbon, nitrogen, and phosphorus, to evaluate their effect on the removal of diclofenac. We also evaluated the photodegradation of diclofenac under the same conditions. The diclofenac removed from the media ranged from 59 to 92%, obtaining the highest removal with S. obliquus. The diclofenac adsorbed on the cell walls ranged from 12.2 to 26.5%, obtaining the highest adsorption with S. obliquus. The diclofenac degraded by light ranged from 15 to 28%. The nutrient deficit showed no influence on the removal of diclofenac in any of the microalgae under study. These results indicate that S. obliquus is the best alternative for the bioremediation of diclofenac. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03268-2.

5.
Plants (Basel) ; 11(11)2022 May 29.
Artículo en Inglés | MEDLINE | ID: mdl-35684220

RESUMEN

The Typha genus comprises plant species extensively studied for phytoremediation processes. Recently, Pseudomonas rhodesiae GRC140, an IAA-producing bacterium, was isolated from Typha latifolia roots. This bacterium stimulates the emergence of lateral roots of Arabidopsis thaliana in the presence and absence of cadmium. However, the bacterial influence on cadmium accumulation by the plant has not been determined. Moreover, the P. rhodesiae GRC140 effect in Cd phytoextraction by T. latifolia remains poorly understood. In this work, an axenic hydroponic culture of T. latifolia was established. The plants were used to evaluate the effects of cadmium stress in axenic plants and determine the effects of P. rhodesiae GRC140 and exogenous indole acetic acid (IAA) on Cd tolerance and Cd uptake by T. latifolia. Biomass production, total chlorophyll content, root electrolyte leakage, catalase activity, total glutathione, and Cd content were determined. The results showed that Cd reduces shoot biomass and increases total glutathione and Cd content in a dose-dependent manner in root tissues. Furthermore, P. rhodesiae GRC140 increased Cd translocation to the shoots, while IAA increased the Cd accumulation in plant roots, indicating that both treatments increase Cd removal by T. latifolia plants. These results indicate that axenic plants in hydroponic systems are adequate to evaluate the Cd effects in plants and suggest that T. latifolia phytoextraction abilities could be improved by P. rhodesiae GRC140 and exogenous IAA application.

6.
Arch Microbiol ; 204(7): 364, 2022 Jun 04.
Artículo en Inglés | MEDLINE | ID: mdl-35661269

RESUMEN

The effect that the fructans of Cichorium intybus and Agave salmiana have on health, as well as on the growth of some Lactobacillus species, has been demonstrated. The aim of this work was to evaluate the effect of linear and branched fructans on the growth of seven strains and some probiotic characteristics. The molecular identification of seven strains was performed. Moreover, the growth, resistance to antibiotics and simulated gastrointestinal conditions were also evaluated when these microorganisms were grown in a culture medium containing agave and chicory fructans. The strains were identified as Lactiplantibacillus plantarum, Lactiplantibacillus pentosus, Lactiplantibacillus fabifermentans and Lactiplantibacillus paraplantarum. The results suggest that the seven Lactobacillus strains were able to grow using agave (branched) and chicory (linear) fructans. The linear and branched fructans statistically influenced the kinetic parameters. The specific growth rate varied between 0.270 and 0.573 h-1 and the generation time between 1.21 and 2.45 h for all strains and culture media. All strains showed a growth of 9 Log CFU/mL in all the culture media. Production of lactic, acetic, propionic, butyric, formic and succinic acid was influenced by linear and branched fructans (p < 0.05). All the strains survived simulated gastrointestinal conditions greater than 83%. The resistance of Lactobacillus against ciprofloxacin and rifaximin was significantly affected by linear and branched fructans, but survival to gastrointestinal conditions was not affected by the type of substrate. These results highlight the use of the seven strains, which have probiotic potential; therefore, these could be applied in several biotechnological products.


Asunto(s)
Agave , Probióticos , Agave/química , Bebidas , Medios de Cultivo , Fructanos/química , Lactobacillus , México
7.
J Gen Intern Med ; 37(3): 624-631, 2022 02.
Artículo en Inglés | MEDLINE | ID: mdl-34993853

RESUMEN

BACKGROUND: Coronavirus disease 2019 (COVID-19) causes a mild illness in most cases; forecasting COVID-19-associated mortality and the demand for hospital beds and ventilators are crucial for rationing countries' resources. OBJECTIVE: To evaluate factors associated with the severity of COVID-19 in Mexico and to develop and validate a score to predict severity in patients with COVID-19 infection in Mexico. DESIGN: Retrospective cohort. PARTICIPANTS: We included 1,435,316 patients with COVID-19 included before the first vaccine application in Mexico; 725,289 (50.5%) were men; patient's mean age (standard deviation (SD)) was 43.9 (16.9) years; 21.7% of patients were considered severe COVID-19 because they were hospitalized, died or both. MAIN MEASURES: We assessed demographic variables, smoking status, pregnancy, and comorbidities. Backward selection of variables was used to derive and validate a model to predict the severity of COVID-19. KEY RESULTS: We developed a logistic regression model with 14 main variables, splines, and interactions that may predict the probability of COVID-19 severity (area under the curve for the validation cohort = 82.4%). CONCLUSIONS: We developed a new model able to predict the severity of COVID-19 in Mexican patients. This model could be helpful in epidemiology and medical decisions.


Asunto(s)
COVID-19 , Hospitalización , Humanos , Masculino , México/epidemiología , Pronóstico , Estudios Retrospectivos , Factores de Riesgo , SARS-CoV-2
8.
Front Plant Sci ; 12: 641420, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34054890

RESUMEN

This year, a respiratory virus caused an emergency pandemic alert in health services around the world, showing the need for biotechnological approaches to fight these diseases. The influenza virus is one of the main viral agents that generate pandemic outbreaks. Currently, the majority of co-circulating influenza A virus (IAV) strains are adamantine- and oseltamivir-resistant strains, and the challenge is to find new antivirals for more efficient treatments. The antiviral entry blocker (EB) peptide is a promising candidate for blocking the virus entry into cells. The aim of this research was to express the EB peptide in the microalgae Chlamydomonas reinhardtii and test its antiviral activity against IAV in vitro. The EB peptide nucleotide sequence was introduced into the nuclear genome of microalgae using Agrobacterium tumefaciens transformation. The EB peptide amount produced in transformed microalgae was 4.99 ± 0.067% of the total soluble protein. In hemagglutination inhibition assays using influenza A/H1N1 pdm and influenza A H1N1/Virginia/ATCC/2009 strains, we reported that the EB peptide extract from the microalgae showed 100-fold higher efficiency than the EB synthetic peptide. In addition, both the EB peptide extract and synthetic peptide inhibited viral replication in MDCK cells (IC50 = 20.7 nM and IC50 = 754.4 nM, respectively); however, the EB peptide extract showed a 32-fold higher antiviral effectiveness than the synthetic peptide against influenza A/H1N1 pdm. Extracts from untransformed and transformed microalgae and synthetic peptide did not show cytotoxic effect on MDCK cell monolayers. Thus, C. reinhardtii may be a fast, safe, and effective expression platform for production of peptides with significant antiviral activity and can be used as a prophylactic treatment to reduce viral propagation.

9.
Biotechnol Prog ; 35(6): e2891, 2019 11.
Artículo en Inglés | MEDLINE | ID: mdl-31374159

RESUMEN

pH variations influence the delivery of essential nutrients and CO2 solubility, which impact algae metabolism. In this study the microalgal growth and chlorophyll, lipid, and fatty acids content; along with the expression of some genes implicated in the biosynthesis of lipids were examined in Chlamydomonas reinhardtii subjected to pH values of 7.0, 7.8, and 8.5. At pH 7.8 an increase in cell growth was observed with a significant accumulation of chlorophyll (1.75-fold) when compared with growth at pH 7, while at pH 8.5 a sharp decrease in both parameters was observed when compared with the other pH values tested. Lipid content increased 3.0 (14.81% of dry cell weight, dcw) and 2.3 times (11.43% dcw) at pH 7.8 and 8.5, respectively, when compared with the experiment at pH 7 (4.97% dcw). The compositions of major fatty acids in the strains growing at pH 7.0, 7.8, or 8.5 were 25.7, 28.0, and 32.1% for palmitic acid; 17.3, 14.7, and 25.7% for oleic acid; and 9.8, 12.1, and 4.6% for linoleic acid; respectively. qRT-PCR analysis showed that the transcripts of ß-carboxyltransferase, Acyl carrier protein 1, acyl-ACP thiolase 1, acyl-sn-glycerol-3-phosphate acyltransferase, and diacylglycerol acyl transferase isoform 3 were significantly induced at pH 7.8 when compared with the other two pH conditions. These results indicate that the induction of genes implicated in the early and final steps of lipid biosynthesis contributes to their accumulation in the stationary phase. Our research suggests that a pH of 7.8 might be ideal to maximize growth and lipid accumulation.


Asunto(s)
Chlamydomonas reinhardtii/metabolismo , Ácidos Grasos/metabolismo , Metabolismo de los Lípidos , Adenosina Trifosfato/biosíntesis , Chlamydomonas reinhardtii/crecimiento & desarrollo , Clorofila/análisis , Ácidos Grasos/análisis , Concentración de Iones de Hidrógeno
10.
J Biotechnol ; 240: 76-84, 2016 Dec 20.
Artículo en Inglés | MEDLINE | ID: mdl-27816654

RESUMEN

In this study, we developed a transplastomic C. reinhardtii strain that accumulates anti-hypertensive peptides. Tandem repeats of VLPVP peptide were included. PCR analysis confirmed the presence of the transgene in the modified strains. After in vitro digestion of biomass of a recombinant C. reinhardtii strain the VLVPV peptide was identified and quantified by HPLC. The highest expression line produced 0.292mg of recombinant protein per mg of freeze-dried biomass. Intragastric administration of the genetically modified strain to spontaneous hypertensive rats at a dose of 30mg/kg of body weight of recombinant protein significantly reduced systolic blood pressure. At the same dose, the recombinant protein exerts an ACE-inhibitory effect. This is the first study that indicates the potential of this microalga producing an antihypertensive peptide as a dietary supplement for hypertension patients.


Asunto(s)
Antihipertensivos/farmacología , Chlamydomonas reinhardtii , Hipertensión/tratamiento farmacológico , Oligopéptidos/farmacología , Proteínas Recombinantes/farmacología , Inhibidores de la Enzima Convertidora de Angiotensina , Animales , Presión Sanguínea/efectos de los fármacos , Hipertensión/fisiopatología , Masculino , Oligopéptidos/biosíntesis , Oligopéptidos/genética , Oligopéptidos/aislamiento & purificación , Ratas , Ratas Endogámicas SHR , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación
11.
World J Microbiol Biotechnol ; 32(4): 55, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26925617

RESUMEN

Chlamydomonas reinhardtii is able to accumulate large amounts of triacylglycerides, the major feedstock for biodiesel production, when grown under stress conditions. In order to characterize gene transcripts induced under nitrogen, iron, and sulfur deprivation in C. reinhardtii; 583 expressed sequence tags (ESTs) were generated through a cDNA library. These sequences were subjected to contig assembly resulting in 30 contigs and 76 singletons. The comparison of the ESTs obtained with public databases allowed to assign putative functions to 66.7 % of the sequences. An important group of the identified genes are related to the lipid metabolic process. A phylogenetic analysis of these sequences identified five isoforms of diacylglycerol O-acyltransferase type 2 (DGAT-2). These genes were selected to measure their relative expression under these stress conditions by means of qRT-PCR. According to the results, the accumulation of DGTT1 mRNA increases considerably under nitrogen and iron inanition when compared to the other isoforms, which indicated that each isoform participates at different levels under each stress condition. These results can help to identify potential genes to be overexpressed by genetic engineering in C. reinhardtii.


Asunto(s)
Chlamydomonas reinhardtii/genética , Diacilglicerol O-Acetiltransferasa/genética , Proteínas de Plantas/genética , Estrés Fisiológico , Triglicéridos/biosíntesis , Chlamydomonas reinhardtii/enzimología , Regulación de la Expresión Génica de las Plantas , Biblioteca de Genes , Hierro/metabolismo , Nitrógeno/metabolismo , Filogenia , ARN Mensajero/genética , ARN de Planta/genética , Azufre/metabolismo
12.
Micron ; 78: 33-39, 2015 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-26218801

RESUMEN

AIM: To determinate the significance of risk factors with the presence of biofilm on catheters of patients attended at tertiary hospital cares. MATERIAL AND METHODS: A total of 126 patients were included, data collection by observing the handling of the CVC, clinical history and microbiological isolation methods of CVCs tips (Roll-plate, sonication and scanning electron microscopy) were evaluated. RESULTS: Certain factors, such as the lack of proper hand washing, the use of primary barriers and preparing medications in the same hospital service, showed an important relationship between biofilm formation in CVCs. The sonication method presented that most of the samples had isolation of multispecies 29 samples (64%); in contrast with the roll-plate method, just one sample (3%) was isolated. CONCLUSIONS: The importance of the strict aseptic techniques of insertion and of the handlings of CVC was highlighted, the failure of both techniques was related to the biofilm formation and was evidenced using the scanning electron microscopy. Since this tool is not available in most hospitals, we present the correlation of those evidences with other standard microbiological methods and risk factors, which are necessary for the sensible detection of the different steps of the biofilm formation on CVC and their correct interpretation with clinical evidences.


Asunto(s)
Biopelículas , Catéteres Venosos Centrales/microbiología , Centros de Atención Terciaria , Biopelículas/crecimiento & desarrollo , Femenino , Humanos , Masculino , Técnicas Microbiológicas , Microscopía Electrónica de Rastreo , Persona de Mediana Edad , Estudios Prospectivos , Factores de Riesgo , Sonicación
13.
Biotechnol Lett ; 37(1): 41-54, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25214222

RESUMEN

Elevated concentrations of cholesterol in plasma are associated with increased risk for heart diseases in humans. Bioactive peptides can be considered as an option to prevent or treat this condition. Currently, there are wide sources of bioactive peptides with hypocholesterolemic activities; however, most researches are focused in bioactive peptides derived from soybean and milk protein. Although there are several preparation methods for these peptides, it is a novel process to prepare bioactive peptides by genetic engineering techniques. In this review, after a general introduction on approaches and advances in bioactive peptides, recombinant strategies to generate hypocholesterolemic peptides and their purification are discussed as well as their application in food and drug design.


Asunto(s)
Anticolesterolemiantes , Péptidos , Proteínas Recombinantes , Anticolesterolemiantes/química , Anticolesterolemiantes/aislamiento & purificación , Anticolesterolemiantes/metabolismo , Proteínas en la Dieta , Escherichia coli , Ingeniería Genética , Péptidos/química , Péptidos/aislamiento & purificación , Péptidos/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo
14.
J Biomed Inform ; 53: 405-14, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25464113

RESUMEN

Exploitation of recombinant DNA and sequencing technologies has led to a new concept in vaccination in which isolated epitopes, capable of stimulating a specific immune response, have been identified and used to achieve advanced vaccine formulations; replacing those constituted by whole pathogen-formulations. In this context, bioinformatics approaches play a critical role on analyzing multiple genomes to select the protective epitopes in silico. It is conceived that cocktails of defined epitopes or chimeric protein arrangements, including the target epitopes, may provide a rationale design capable to elicit convenient humoral or cellular immune responses. This review presents a comprehensive compilation of the most advantageous online immunological software and searchable, in order to facilitate the design and development of vaccines. An outlook on how these tools are supporting vaccine development is presented. HIV and influenza have been taken as examples of promising developments on vaccination against hypervariable viruses. Perspectives in this field are also envisioned.


Asunto(s)
Biología Computacional/métodos , Mapeo Epitopo/métodos , Vacunas/química , Algoritmos , Sistemas de Computación , ADN/química , Epítopos/química , Genoma , Infecciones por VIH/diagnóstico , Humanos , Vacunas contra la Influenza/inmunología , Gripe Humana/diagnóstico , Gripe Humana/inmunología , Informática Médica , Proteínas/química , Proteínas Recombinantes/química , Programas Informáticos , Linfocitos T/inmunología
15.
J Biotechnol ; 184: 27-38, 2014 Aug 20.
Artículo en Inglés | MEDLINE | ID: mdl-24844864

RESUMEN

The high demand for less polluting, newer, and cheaper fuel resources has increased the search of the most innovative options for the production of the so-called biofuels. Chlamydomonas reinhardtii is a photosynthetic unicellular algae with multiple biotechnological advantages such as easy handling in the laboratory, a simple scale-up to industrial levels, as well as a feasible genetic modification at nuclear and chloroplast levels. Besides, its fatty acids can be used to produce biofuels. Previous studies in plants have found that the over expression of DOF-type transcription factor genes increases the synthesis and the accumulation of total lipids in seeds. In this context, the over-expression of a DOF-type transcription factor in C. reinhardtii was applied as approach to increase the amount of lipids. The results indicate higher amounts (around 2-fold) of total lipids, which are mainly fatty acids, in the genetically C. reinhardtii modified strains when compared with the non-genetically modified strain. In order to elucidate the possible function of the introduced Dof-type transcription factor, we performed a transcription profile of 8 genes involved in fatty acid biosynthesis and 6 genes involved in glycerolipid biosynthesis, by quantitative real time (qRT-PCR). Differential expression profile was observed, which can explain the increase in lipid accumulation. However, these strains did not show notable changes in the fatty acid profile. This work represents an early effort in generating a strategy to increase fatty acids production in C. reinhardtii and their use in biofuel synthesis.


Asunto(s)
Biocombustibles , Cloroplastos/metabolismo , Metabolismo de los Lípidos , Lípidos/biosíntesis , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/metabolismo , Cloroplastos/genética , Regulación de la Expresión Génica de las Plantas , Lípidos/genética , Fotosíntesis/genética , Semillas/genética , Semillas/metabolismo , Factores de Transcripción/biosíntesis
16.
Plant Mol Biol ; 78(4-5): 337-49, 2012 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-22228408

RESUMEN

Although the human immunodeficiency virus (HIV) causes one of the most important infectious diseases worldwide, attempts to develop an effective vaccine remain elusive. Designing recombinant proteins capable of eliciting significant and protective mammalian immune responses remain a priority. Moreover, large-scale production of proteins of interest at affordable cost remains a challenge for modern biotechnology. In this study, a synthetic gene encoding a C4V3 recombinant protein, known to induce systemic and mucosal immune responses in mammalian systems, has been introduced into tobacco chloroplasts to yield high levels of expression. Integration of the transgene into the tobacco plastome has been verified by Southern blot hybridization. The recombinant C4V3 protein is also detected in tobacco chloroplasts by confocal microscopy. Reactivity of the heterologous protein with both an anti-C4V3 rabbit serum as well as sera from HIV positive patients have been assayed using Western blots. When administered by the oral route in a four-weekly dose immunization scheme, the plant-derived C4V3 has elicited both systemic and mucosal antibody responses in BALB/c mice, as well as CD4+ T cell proliferation responses. These findings support the viability of using plant chloroplasts as biofactories for HIV candidate vaccines, and could serve as important vehicles for the development of a plant-based candidate vaccine against HIV.


Asunto(s)
Fármacos Anti-VIH/inmunología , Cloroplastos/genética , Proteína gp120 de Envoltorio del VIH/inmunología , Fragmentos de Péptidos/inmunología , Péptidos/administración & dosificación , Péptidos/inmunología , Vacunas Sintéticas/administración & dosificación , Administración Oral , Animales , Fármacos Anti-VIH/administración & dosificación , Cloroplastos/inmunología , Femenino , Proteína gp120 de Envoltorio del VIH/genética , Seropositividad para VIH , Humanos , Inmunidad Mucosa/inmunología , Inmunización , Ratones , Ratones Endogámicos BALB C , Fragmentos de Péptidos/genética , Péptidos/genética , Plantas Modificadas Genéticamente , Conejos , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacología , Nicotiana/genética
17.
Plant Cell Rep ; 31(3): 479-94, 2012 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-22080228

RESUMEN

Chlamydomonas reinhardtii has many advantages compared with traditional systems for the molecular farming of recombinant proteins. These include low production costs, rapid scalability at pilot level, absence of human pathogens and the ability to fold and assemble complex proteins accurately. Currently, the successful expression of several proteins with pharmaceutical relevance has been reported from the nuclear and the chloroplastic genome of this alga, demonstrating its usefulness for biotechnological applications. However, several factors affect the level of recombinant protein expression in Chlamydomonas such as enhancer elements, codon dependency, sensitivity to proteases and transformation-associated genotypic modification. The present review outlines a number of strategies to increase protein yields and summarizes recent achievements in algal protein production including biopharmaceuticals such as vaccines, antibodies, hormones and enzymes with implications on health-related approaches. The current status of bioreactor developments for algal culture and the challenges of scale-up and optimization processes are also discussed.


Asunto(s)
Chlamydomonas reinhardtii/metabolismo , Industria Farmacéutica/métodos , Microalgas/metabolismo , Proteínas Recombinantes/biosíntesis , Reactores Biológicos , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/crecimiento & desarrollo , Enzimas/biosíntesis , Hormonas/biosíntesis , Humanos , Microalgas/genética , Microalgas/crecimiento & desarrollo , Agricultura Molecular , Planticuerpos/metabolismo , Plantas Modificadas Genéticamente , Vacunas
18.
Plant Cell Rep ; 30(8): 1367-82, 2011 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-21505834

RESUMEN

Genetic engineering revolutionized the concept of traditional vaccines since subunit vaccines became reality. Additionally, over the past two decades plant-derived antigens have been studied as potential vaccines with several advantages, including low cost and convenient administration. More specifically, genetic fusions allowed the expression of fusion proteins carrying two or more components with the aim to elicit immune responses against different targets, including antigens from distinct pathogens or strains. This review aims to provide an update in the field of the production of plant-based vaccine, focusing on those approaches based on the production of chimeric proteins comprising antigens from human pathogens, emphasizing the case of cholera toxin/E. coli enterotoxin fusions, chimeric viruses like particles approaches as well as the possible use of adjuvant-producing plants as expression hosts. Challenges for the near future in this field are also discussed.


Asunto(s)
Ingeniería Genética/métodos , Plantas Modificadas Genéticamente/metabolismo , Vacunas/biosíntesis , Adyuvantes Inmunológicos/química , Toxina del Cólera/biosíntesis , Toxina del Cólera/inmunología , Enterotoxinas/biosíntesis , Enterotoxinas/inmunología , Ingeniería Genética/tendencias , Plantas Modificadas Genéticamente/inmunología , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/inmunología
19.
Plant Cell Rep ; 30(6): 1145-52, 2011 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-21318355

RESUMEN

Enterotoxigenic Escherichia coli (ETEC) is one of the main causative agents of diarrhea in infants and for travelers. Inclusion of a heat-stable (ST) toxin into vaccine formulations is mandatory as most ETEC strains can produce both heat-labile (LT) and ST enterotoxins. In this study, a genetic fusion gene encoding for an LTB:ST protein has been constructed and transferred into tobacco via Agrobacterium tumefaciens-mediated transformation. Transgenic tobacco plants carrying the LTB:ST gene are then subjected to GM1-ELISA revealing that the LTB:ST has assembled into pentamers and displays antigenic determinants from both LTB and ST. Protein accumulation of up to 0.05% total soluble protein is detected. Subsequently, mucosal and systemic humoral responses are elicited in mice orally dosed with transgenic tobacco leaves. This has suggested that the plant-derived LTB:ST is immunogenic via the oral route. These findings are critical for the development of a plant-based vaccine capable of eliciting broader protection against ETEC and targeting both LTB and ST. Features of this platform in comparison to transplastomic approaches are discussed.


Asunto(s)
Toxinas Bacterianas/metabolismo , Núcleo Celular/metabolismo , Enterotoxinas/metabolismo , Escherichia coli/metabolismo , Nicotiana/genética , Subunidades de Proteína/metabolismo , Proteínas Recombinantes de Fusión/inmunología , Administración Oral , Secuencia de Aminoácidos , Animales , Formación de Anticuerpos/inmunología , Antígenos/inmunología , Secuencia de Bases , Proteínas de Escherichia coli , Ratones , Datos de Secuencia Molecular , Plantas Modificadas Genéticamente , Reacción en Cadena de la Polimerasa , Proteínas Recombinantes de Fusión/administración & dosificación , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética
20.
J Plant Physiol ; 168(2): 174-80, 2011 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-20655621

RESUMEN

Expression of the protective F1 and V antigens of Yersinia pestis, as a fusion protein, in carrot was pursued in an effort to develop an alternative vaccine production system against the serious plague disease. Transgenic carrot plants carrying the F1-V encoding gene were developed via Agrobacterium-mediated transformation. Presence, integration, and expression of the F1-V encoding gene were confirmed by polymerase chain reaction (PCR), DNA gel blot analysis, and reverse-transcriptase (RT)-PCR analyses, respectively. An ELISA assay confirmed the antigenicity of the plant-derived F1-V fusion protein. Immunogenicity was evaluated subcutaneously in mice using a soluble protein extract of freeze-dried transgenic carrot. Significant antibody levels were detected following immunization. These results demonstrated that the F1-V protein could be expressed in carrot tap roots, and that the carrot F1-V recombinant protein retained its antigenicity and immunogenicity.


Asunto(s)
Antígenos Bacterianos/metabolismo , Proteínas Bacterianas/metabolismo , Daucus carota/metabolismo , Raíces de Plantas/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Proteínas Citotóxicas Formadoras de Poros/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Yersinia pestis/metabolismo , Animales , Antígenos Bacterianos/genética , Proteínas Bacterianas/genética , Daucus carota/genética , Ensayo de Inmunoadsorción Enzimática , Ratones , Ratones Endogámicos BALB C , Peste/inmunología , Vacuna contra la Peste/genética , Vacuna contra la Peste/inmunología , Vacuna contra la Peste/metabolismo , Raíces de Plantas/genética , Plantas Modificadas Genéticamente/genética , Proteínas Citotóxicas Formadoras de Poros/genética , Proteínas Recombinantes de Fusión/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Yersinia pestis/genética
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