14 results on '"Juan C, Cancino-Diaz"'
Search Results
2. Assessment of the tolerance to Fe, Cu and Zn of a sulfidogenic sludge generated from hydrothermal vents sediments as a basis for its application on metals precipitation
- Author
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Alberto Ordaz, Janet Jan-Roblero, Claudia Guerrero-Barajas, Khemlal Nirmalkar, Paola Zárate-Segura, Juan C. Cancino-Diaz, and Jaime García-Mena
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0301 basic medicine ,Geologic Sediments ,Sulfide ,Iron ,03 medical and health sciences ,chemistry.chemical_compound ,Hydrothermal Vents ,0302 clinical medicine ,Bioremediation ,Genetics ,Desulfovibrio desulfuricans ,Sulfate ,Sulfate-reducing bacteria ,Molecular Biology ,Biological Oxygen Demand Analysis ,chemistry.chemical_classification ,Bacteria ,Sewage ,biology ,Chemistry ,Chemical oxygen demand ,General Medicine ,biology.organism_classification ,Desulfovibrio ,Zinc ,030104 developmental biology ,Peptococcaceae ,030220 oncology & carcinogenesis ,Environmental chemistry ,Desulfotomaculum ,Microcosm ,Copper - Abstract
A paramour factor limiting metal-microorganism interaction is the metal ion concentration, and the metal precipitation efficiency driven by microorganisms is sensitive to metal ion concentration. The aim of the work was to determine the tolerance of the sulfidogenic sludge generated from hydrothermal vent sediments at microcosms level to different concentrations of Fe, Cu and Zn and the effect on the microbial community. In this study the chemical oxygen demand (COD) removal, sulfate-reducing activity (SRA) determination, inhibition effect through the determination of IC50, and the characterization of the bacterial community´s diversity were conducted. The IC50 on SRA was 34 and 81 mg/L for Zn and Cu, respectively. The highest sulfide concentration (H2S mg/L) and % of sulfate reduction obtained were: 511.30 ± 0.75 and 35.34 ± 0.51 for 50 mg/L of Fe, 482.48 ± 6.40 and 33.35 ± 0.44 for 10 mg/L of Cu, 442.26 ± 17.1 and 30.57 ± 1.18 for 10 mg/L of Zn, respectively. The COD removal rates were of 71.81 ± 7.6, 53.92 ± 1.07 and 57.68 ± 10.2 mg COD/ L d for Fe (50 mg/L), Cu (40 mg/L) and Zn (20 mg/L), respectively. Proteobacteria, Firmicutes, Chloroflexi and Actinobacteria were common phyla to four microcosms (stabilized sulfidogenic and added with Fe, Cu or Zn). The dsrA genes of Desulfotomaculum acetoxidans, Desulfotomaculum gibsoniae and Desulfovibrio desulfuricans were expressed in the microcosms supporting the SRA results. The consortia could be explored for ex-situ bioremediation purposes in the presence of the metals tested in this work.
- Published
- 2020
3. Genotypic and phenotypic changes of Staphylococcus epidermidis during relapse episodes in prosthetic joint infections
- Author
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Alejandra Aquino-Andrade, Ashutosh Sharma, Gabriel Betanzos-Cabrera, Rafael Franco-Cendejas, Mario E. Cancino-Diaz, Juan C. Cancino-Diaz, Sandra Rodríguez-Martínez, and Silvestre Ortega-Peña
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Adult ,Male ,medicine.medical_specialty ,Prosthesis-Related Infections ,Genotype ,Virulence Factors ,Joint Prosthesis ,Clone (cell biology) ,Microbial Sensitivity Tests ,Microbiology ,03 medical and health sciences ,Antibiotic resistance ,Medical microbiology ,Recurrence ,Staphylococcus epidermidis ,Drug Resistance, Multiple, Bacterial ,Media Technology ,medicine ,Pulsed-field gel electrophoresis ,Humans ,Aged ,030304 developmental biology ,0303 health sciences ,biology ,030306 microbiology ,Correction ,Middle Aged ,Staphylococcal Infections ,biology.organism_classification ,Antimicrobial ,Anti-Bacterial Agents ,Cross-Sectional Studies ,Phenotype ,Biofilms ,Vancomycin ,medicine.drug - Abstract
Staphylococcus epidermidis is a coagulase-negative bacterium capable of causing recurrent relapses in prosthetic joint infection (PJI). The aim of this study was to determine if Staphylococcus epidermidis isolates from patients with recurrent relapses of prosthetic joint infection (PJI) changed genotypically (pulsed-field gel electrophoresis (PFGE) pattern analysis and genes involved in biofilm formation) and phenotypically (antimicrobial resistance, biofilm formation) during the different episodes. Four patients with PJI recurrent relapses were evaluated clinically and microbiologically. Genotypic and phenotypic characteristics of 31 S. epidermidis isolates were determined. In all cases, PJI was treated with antimicrobial therapy and resection of the prosthesis without reimplantation. Months later, all patients had a relapse episode and treated with rifampin plus vancomycin and surgical debridement. Changes in the antibiotics resistance profile in isolates from patients 1 and 2 were observed in the two episodes. Patient 1 had four clones A, B, C, and D that were distributed differentially in the two episodes. Similarly, patients 2 and 3 had two clones and subclones (E-E1 and F-F1, respectively), and patient 4 had only the clone G in both episodes. The clone F formed small-colony variants (SCVs). High level of biofilm formation was found in all clones, except for clones D and G. Clones/subclones showed a genotypic variation in icaA, sdrF, bap, sesI, and embp genes. The principal coordinate analysis showed that all clones/subclones were different. These results showed that the initial infective clone of S. epidermidis from PJI, changed genotypically and phenotypically after a second relapse as a response to the treatment.
- Published
- 2019
4. Overview of Staphylococcus epidermidis cell wall-anchored proteins: potential targets to inhibit biofilm formation
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Silvestre Ortega-Peña, Mario E. Cancino-Diaz, Sandra Rodríguez-Martínez, Sergio Martínez-García, and Juan C. Cancino-Diaz
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0301 basic medicine ,medicine.drug_class ,medicine.medical_treatment ,Antibiotics ,Microbiology ,Cell wall ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Bacterial Proteins ,Cell Wall ,Staphylococcus epidermidis ,Drug Discovery ,Genetics ,medicine ,Humans ,Molecular Biology ,biology ,Chemistry ,Biofilm ,General Medicine ,Immunotherapy ,Staphylococcal Infections ,biochemical phenomena, metabolism, and nutrition ,Antimicrobial ,biology.organism_classification ,Anti-Bacterial Agents ,030104 developmental biology ,Biofilms ,030220 oncology & carcinogenesis - Abstract
Currently, the treatment of infections by Staphylococcus epidermidis (S. epidermidis) represents a challenge because some strains have multidrug-resistance to antimicrobial products (antibiotic and biocides) and can produce biofilms. These biofilms protect bacterial cells from both antimicrobials and the host immune response. Therefore, it is crucial to encourage research on the development of new treatments. One method is immunotherapy, targeting components of S. epidermidis, such as S. epidermidis surface (Ses) proteins. Ses is expressed constitutively in most strains, and they participate in biofilm formation. This review is an update on Ses, regarding their structure, biological function, their relationship with S. epidermidis biofilm formation, and its possible role as therapeutic targets to develop immunotherapeutic treatments to prevent infections by S. epidermidis.
- Published
- 2019
5. Differential Expression of the apsXRS System by Antimicrobial Peptide LL-37 in Commensal and Clinical Staphylococcus epidermidis Isolates
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Miguel Ibáñez-Hernández, Sergio Martínez-García, Erika T. Quintana, Mario E. Cancino-Diaz, Rodolfo Marsch-Moreno, Marisa Cruz-Aguilar, Juan Carlos Zenteno, Cipriano Chávez-Cabrera, Sandra Rodríguez-Martínez, Norma Velázquez-Guadarrama, Juan C. Cancino-Diaz, and Gabriel Betanzos-Cabrera
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0106 biological sciences ,0303 health sciences ,medicine.medical_specialty ,Strain (chemistry) ,biology ,030306 microbiology ,Operon ,Promoter ,Antimicrobial ,biology.organism_classification ,01 natural sciences ,Microbiology ,Genome ,03 medical and health sciences ,Medical microbiology ,Staphylococcus epidermidis ,010608 biotechnology ,medicine ,Gene - Abstract
The three-component apsXRS system senses and responds to cationic antimicrobial peptides (CAMPs), which induces the expression of the dlt operon and the genes mprF and vrafG, modifying the surface net charge in Staphylococcus epidermidis, resulting in the repulsion of CAMPs. The apsXRS system has been only studied in the S. epidermidis 1457 strain, and there are no studies of prevalence and level of expression of apsXRS in commensal and clinical isolates. From 60 isolates, those selected from commensal healthy skin (n = 20), commensal healthy conjunctive (n = 10), and clinical ocular infection (n = 30) presented the apsX, apsR, and apsS genes in their genomes. Constitutive expression of apsX, apsR, and apsS genes was determined by RT-qPCR in all isolates. It was found that expression of apsX, apsR, and apsS was 3.3–5.9-fold higher in commensal isolates stimulated with LL-37 (15 µg/mL) than in clinical isolates. Similarly, expression of the dlt operon and the genes mprF, and vraFG was 8–10-fold higher in commensal isolates than in clinical. However, LL-37 did not increase the addition of lysine in the phospholipids of the cytoplasmic membrane in any of the isolates. Mutations in the apsS loop region, apsR, and their promoter sequence were not found. These results demonstrated that apsXRS system is essential in all isolates for its constitutive expression; however, LL-37 caused an increase of apsXRS expression in commensal isolates, suggesting that S. epidermidis isolates do not respond in the same way to the presence of LL-37.
- Published
- 2019
6. Construction of a synthetic protein using PCR with a high essential amino acid content for nutritional purposes
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Ma I Sánchez-Crisóstomo, Juan C. Cancino-Diaz, Eduardo Madrigal-Santillán, M I Rojo-López, Ashutosh Sharma, Gabriel Betanzos-Cabrera, H Jaimes-Díaz, and José Alberto Ariza-Ortega
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0301 basic medicine ,Recombinant Fusion Proteins ,Protein Engineering ,medicine.disease_cause ,Polymerase Chain Reaction ,Chromatography, Affinity ,03 medical and health sciences ,0302 clinical medicine ,Affinity chromatography ,Escherichia coli ,Genetics ,medicine ,Overlap extension polymerase chain reaction ,Cloning, Molecular ,Molecular Biology ,Essential amino acid ,chemistry.chemical_classification ,biology ,Chemistry ,Proteins ,Biological value ,General Medicine ,Protein engineering ,Recombinant Proteins ,Amino acid ,Ovalbumin ,030104 developmental biology ,Biochemistry ,030220 oncology & carcinogenesis ,Dietary Supplements ,biology.protein ,Amino Acids, Essential - Abstract
Ovalbumin is considered a protein of high nutritional value because it contains essential amino acids and is highly digestible. Therefore, it has a high biological value. Currently, the high food demand requires worldwide attention because food production is insufficient. Therefore, other alternatives are necessary to satisfy food demands, such as protein engineering. In this work, a protein with a high essential amino acid content similar to ovalbumin was synthesized by protein engineering, expressed, and digested in vitro. The assembly and sequential overlap extension PCR strategy was used to synthesize a 345-bp gene that encodes a high essential amino acid content protein (HEAAP). The 345-bp product was cloned into the vector pBAD TOPO®, and expressed in Escherichia coli BL21. PCR reactions and sequencing demonstrated the presence, orientation, and correct sequence of the insert. HEAAP expression was induced by L-arabinose and then purified using Ni-NTA affinity chromatography. The expression in E. coli was low and barely detected by Western blot assay. The in vitro multienzyme digestibility of HEAAP was around 79%, which suggests that the protein is potentially nutritious. Virtual analysis classifies the protein as unstable and hydrophilic, with a half-life in E. coli of 10 h. The recombinant HEAAP was successfully synthesized, but it is necessary to improve the digestibility and to optimize expression including selecting other expression models.
- Published
- 2019
7. Competition/antagonism associations of biofilm formation among Staphylococcus epidermidis Agr groups I, II, and III
- Author
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José Martin Murrieta-Coxca, Juan C. Cancino-Diaz, César I. Ortiz-García, Sergio Martínez-García, Marisa Cruz-Aguilar, Juan Carlos Zenteno, Sonia Mayra Pérez-Tapia, Mario E. Cancino-Diaz, and Sandra Rodríguez-Martínez
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DNA, Bacterial ,Genotype ,Operon ,Virulence ,Biology ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,Microbiology ,Mice ,03 medical and health sciences ,Bacterial Proteins ,In vivo ,Staphylococcus epidermidis ,medicine ,Animals ,Humans ,Skin ,030304 developmental biology ,Mice, Inbred BALB C ,0303 health sciences ,030306 microbiology ,Biofilm ,Quorum Sensing ,Gene Expression Regulation, Bacterial ,General Medicine ,Staphylococcal Infections ,biochemical phenomena, metabolism, and nutrition ,bacterial infections and mycoses ,biology.organism_classification ,In vitro ,Disease Models, Animal ,Genes, Bacterial ,Biofilms ,Catheter-Related Infections ,Trans-Activators ,bacteria ,Female ,Antagonism ,Staphylococcus ,Multilocus Sequence Typing - Abstract
Staphylococci have quorum-sensing (QS) systems that enable cell-to-cell communication, as well as the regulation of numerous colonization and virulence factors. The accessory gene regulator (Agr) operon is one of the Staphylococcus genus QS systems. Three groups (I, II, and III) are present in Staphylococcus epidermidis Agr operon. To date, it is unknown whether Agr groups can interact symbiotically during biofilm development. This study analyzed a symbiotic association among Agr groups during biofilm formation in clinical and commensal isolates. Different combinations among Agr group isolates was used to study biofilm formation in vitro and in vivo (using a mouse catheter-infection model). The analysis of Agr groups were also performed from samples of human skin (head, armpits, and nostrils). Different predominant coexistence was found within biofilms, suggesting symbiosis type. In vitro, Agr I had a competition with Agr II and Agr III. Agr II had a competition with Agr III, and Agr II was an antagonist to Agr I and III when the three strains were combined. In vivo, Agr II had a competition to Agr I, but Agr I and II were antagonists to Agr III. The associations found in vitro and in vivo were also found in different sites of the skin. Besides, other associations were observed: Agr III antagonized Agr I and II, and Agr III competed with Agr I and Agr II. These results suggest that, in S. epidermidis, a symbiotic association of competition and antagonism occurs among different Agr groups during biofilm formation.
- Published
- 2019
8. Daily supplementation with fresh pomegranate juice increases paraoxonase 1 expression and activity in mice fed a high-fat diet
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Guadalupe López-Rodríguez, E. Martínez-Hinojosa, Juan C. Cancino-Diaz, Helen Belefant-Miller, Diego Estrada-Luna, and Gabriel Betanzos-Cabrera
- Subjects
0301 basic medicine ,Antioxidant ,medicine.medical_treatment ,Saturated fat ,Gene Expression ,Medicine (miscellaneous) ,030204 cardiovascular system & hematology ,Diet, High-Fat ,Arylesterase ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,medicine ,Animals ,RNA, Messenger ,Food science ,Sodium Cholate ,Triglycerides ,Lythraceae ,Nutrition and Dietetics ,biology ,Aryldialkylphosphatase ,Chemistry ,Cholesterol ,Body Weight ,Paraoxonase ,medicine.disease ,PON1 ,Obesity ,Fruit and Vegetable Juices ,030104 developmental biology ,Liver ,Fruit ,Dietary Supplements ,biology.protein - Abstract
Studies have found that pomegranate juice (PJ) consumption increases the binding of high-density lipoproteins (HDL) to paraoxonase 1 (PON1), thus increasing the catalytic activity of this enzyme. PON1 is an antioxidant arylesterase synthesized in the liver and transported in plasma in association with HDL. Decreased levels of PON1 are associated with higher levels of cholesterol. We determined the effects of PJ on body weight, cholesterol, and triacylglycerols through 5 months of supplementation. In addition, the effect of PJ on pon1 gene expression in the liver was also measured by RT-qPCR as well as the activity in serum by a semiautomated method using paraoxon as a substrate. CD-1 mice were either fed a control diet or were fed a high-fat diet 1.25% (wt/wt) cholesterol, 0.5% (wt/wt) sodium cholate, and 15% (wt/wt) saturated fat. 300 μL of PJ containing 0.35 mmol total polyphenols was administered by oral gavage to half of the high fat mice daily. The rest of the high fat mice and the control mice were administered with 300 μL of water. PJ-supplemented animals had significantly higher levels of expression of pon1 compared to the unsupplemented group. PJ-supplemented animals had twice the PON1 activity of the unsupplemented group. In addition, PJ-supplemented animals had the lowest body weight and significantly reduced cholesterol and triacylglycerol levels, although the tricylglycerol levels were not consistently decreased. These results suggest that PJ protects against the effects of a high-fat diet in body weight, and cholesterol levels.
- Published
- 2017
9. Correction to: Genotypic and phenotypic changes of Staphylococcus epidermidis during relapse episodes in prosthetic joint infections
- Author
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Ashutosh Sharma, Silvestre Ortega-Peña, Sandra Rodríguez-Martínez, Gabriel Betanzos-Cabrera, Juan C. Cancino-Diaz, Mario E. Cancino-Diaz, and Alejandra Aquino-Andrade
- Subjects
medicine.medical_specialty ,Clinical Microbiology - Research Paper ,biology ,Prosthetic joint ,biology.organism_classification ,Microbiology ,Medical microbiology ,Microbial ecology ,Staphylococcus epidermidis ,Genotype ,Media Technology ,medicine ,Food microbiology - Abstract
Staphylococcus epidermidis is a coagulase-negative bacterium capable of causing recurrent relapses in prosthetic joint infection (PJI). The aim of this study was to determine if Staphylococcus epidermidis isolates from patients with recurrent relapses of prosthetic joint infection (PJI) changed genotypically (pulsed-field gel electrophoresis (PFGE) pattern analysis and genes involved in biofilm formation) and phenotypically (antimicrobial resistance, biofilm formation) during the different episodes. Four patients with PJI recurrent relapses were evaluated clinically and microbiologically. Genotypic and phenotypic characteristics of 31 S. epidermidis isolates were determined. In all cases, PJI was treated with antimicrobial therapy and resection of the prosthesis without reimplantation. Months later, all patients had a relapse episode and treated with rifampin plus vancomycin and surgical debridement. Changes in the antibiotics resistance profile in isolates from patients 1 and 2 were observed in the two episodes. Patient 1 had four clones A, B, C, and D that were distributed differentially in the two episodes. Similarly, patients 2 and 3 had two clones and subclones (E-E1 and F-F1, respectively), and patient 4 had only the clone G in both episodes. The clone F formed small-colony variants (SCVs). High level of biofilm formation was found in all clones, except for clones D and G. Clones/subclones showed a genotypic variation in icaA, sdrF, bap, sesI, and embp genes. The principal coordinate analysis showed that all clones/subclones were different. These results showed that the initial infective clone of S. epidermidis from PJI, changed genotypically and phenotypically after a second relapse as a response to the treatment.
- Published
- 2020
10. Evaluation of the Removal of Pyrene and Fluoranthene by Ochrobactrum anthropi, Fusarium sp. and Their Coculture
- Author
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Juan A. Cruz-Maya, Cesar M. Flores-Ortiz, Janet Jan-Roblero, Diana K. Ortega-González, Eliseo Cristiani-Urbina, and Juan C. Cancino-Diaz
- Subjects
Fusarium ,Ochrobactrum anthropi ,Microorganism ,Bioengineering ,Applied Microbiology and Biotechnology ,Biochemistry ,Microbiology ,chemistry.chemical_compound ,Molecular Biology ,Fluoranthene ,Fluorenes ,Pyrenes ,biology ,General Medicine ,biology.organism_classification ,Coculture Techniques ,Culture Media ,Kinetics ,Biodegradation, Environmental ,chemistry ,Pyrene ,Environmental Pollutants ,Antagonism ,Energy source ,Bacteria ,Biotechnology - Abstract
Fluoranthene and pyrene are polycyclic aromatic hydrocarbons of high molecular weight that are recalcitrant and toxic to humans; therefore, their removal from the environment is crucial. From hydrocarbon-contaminated soil, 25 bacteria and 12 filamentous fungi capable of growth on pyrene and fluoranthene as the sole carbon and energy source were isolated. From these isolates, Ochrobactrum anthropi BPyF3 and Fusarium sp. FPyF1 were selected and identified because they grew quickly and abundantly in both hydrocarbons. Furthermore, O. anthropi BPyF3 and Fusarium sp. FPyF1 were most efficient at removing pyrene (50.39 and 51.32 %, respectively) and fluoranthene (49.85 and 49.36 %, respectively) from an initial concentration of 50 mg L(-1) after 7 days of incubation. Based on this and on the fact that there was no antagonism between the two microorganisms, a coculture composed of O. anthropi BPyF3 and Fusarium sp. FPyF1 was formed to remove fluoranthene and pyrene at an initial concentration of 100 mg L(-1) in a removal kinetic assay during 21 days. Fluoranthene removal by the coculture was higher (87.95 %) compared with removal from the individual cultures (68.95 % for Fusarium sp. FPyF1 and 64.59 % for O. anthropi BPyF3). In contrast, pyrene removal by the coculture (99.68 %) was similar to that obtained by the pure culture of Fusarium sp. FPyF1 (99.75 %). The kinetics of removal for both compounds was adjusted to a first-order model. This work demonstrates that the coculture formed by Fusarium sp. FPyF1 and O. anthropi BPyF3 has greater potential to remove fluoranthene than individual cultures; however, pyrene can be removed efficiently by Fusarium sp. FPyF1 alone.
- Published
- 2014
11. Roles of bacterial membrane vesicles
- Author
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Stephen M. Boyle, Minerva Georgina Araiza-Villanueva, Eric Daniel Avila-Calderón, Juan C. Cancino-Diaz, Edgar Oliver López-Villegas, Nammalwar Sriranganathan, and Araceli Contreras-Rodríguez
- Subjects
Virulence Factors ,Antigenic protein ,Virulence ,Gram-Positive Bacteria ,Cell Membrane Structures ,Biochemistry ,Microbiology ,Cell Wall ,Gram-Negative Bacteria ,Genetics ,Animals ,Humans ,Membrane vesicle ,Molecular Biology ,Gram-Positive Bacterial Infections ,biology ,Vesicle ,Cell Membrane ,General Medicine ,biology.organism_classification ,Cell biology ,Acellular vaccines ,Cell envelope ,Gram-Negative Bacterial Infections ,Bacterial outer membrane ,Bacteria - Abstract
Outer membrane vesicles (OMVs) are released from the outer membrane of Gram-negative bacteria. Moreover, Gram-positive bacteria also produce membrane-derived vesicles. As OMVs transport several bacterial components, especially from the cell envelope, their interaction with the host cell, with other bacteria or as immunogens, have been studied intensely. Several functions have been ascribed to OMVs, especially those related to the transport of virulence factors, antigenic protein composition, and development as acellular vaccines. In this work, we review some of the recent findings about OMVs produced by specific pathogenic bacterial species.
- Published
- 2014
12. Adsorption and native microbiota of an agricultural soil are involved in the removal of fluoranthene
- Author
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Juan C. Cancino-Diaz, Alejandro Ponce-Mendoza, Juan A. Cruz-Maya, O. J. Hernández-Hernández, L. R. Evaristo-Vázquez, Héctor Flores-Herrera, Janet Jan-Roblero, Cesar M. Flores-Ortiz, and María Soledad Vásquez-Murrieta
- Subjects
Fluoranthene ,Ecology ,business.industry ,Soil Science ,Soil classification ,Plant Science ,Soil contamination ,chemistry.chemical_compound ,Adsorption ,chemistry ,Agriculture ,Environmental chemistry ,Soil pH ,Cation-exchange capacity ,business ,Microcosm ,Agronomy and Crop Science - Published
- 2016
13. Peptidoglycan and muramyl dipeptide from Staphylococcus aureus induce the expression of VEGF-A in human limbal fibroblasts with the participation of TLR2-NFκB and NOD2-EGFR
- Author
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Sandra Rodríguez-Martínez, Mario E. Cancino-Diaz, Juan C. Cancino-Diaz, and Marco A. Juárez-Verdayes
- Subjects
Vascular Endothelial Growth Factor A ,Staphylococcus aureus ,Time Factors ,Nod2 Signaling Adaptor Protein ,Enzyme-Linked Immunosorbent Assay ,Peptidoglycan ,Limbus Corneae ,Biology ,Microbiology ,Cellular and Molecular Neuroscience ,chemistry.chemical_compound ,NOD2 ,parasitic diseases ,NOD1 ,Humans ,RNA, Messenger ,Eye Proteins ,Receptor ,Cells, Cultured ,Dose-Response Relationship, Drug ,Reverse Transcriptase Polymerase Chain Reaction ,NF-kappa B ,Fibroblasts ,Flow Cytometry ,Molecular biology ,Toll-Like Receptor 2 ,Sensory Systems ,ErbB Receptors ,body regions ,Vascular endothelial growth factor ,Ophthalmology ,TLR2 ,Gene Expression Regulation ,chemistry ,Lipoteichoic acid ,Acetylmuramyl-Alanyl-Isoglutamine ,Muramyl dipeptide - Abstract
Keratitis caused by Staphylococcus aureus often leads to Vascular Endothelial Growth Factor (VEGF)-dependent neovascularization, but contribution of peptidoglycan (PGN), muramyl dipeptide (MDP) and lipoteichoic acid (LTA) from S. aureus to VEGF-dependent neovascularization has not been well-studied. This work was focused on the analysis of S. aureus cell wall components in the production of VEGF family members (VEGF-A, VEGF-B, VEGF-C and VEGF-D) in ocular limbal fibroblasts. Primary culture of human limbal fibroblasts (PCHLFs) were stimulated with PGN, MDP, and LTA, and VEGF family; toll-like receptor 2 (TLR2), nucleotide-binding oligomerization domain 1 (NOD1), and NOD2 expression were determined by RT-PCR. Anti-TLR2 antibody, epidermal growth factor receptor (EGFR) signaling inhibitors (AG1478 and PD98059), and NFκB activation were used to analyze VEGF-A by ELISA. TLR2 and NOD1 expression were analyzed by flow cytometry. The stimulation of PCHLFs with PGN and MDP increased the levels of VEGF-A expression (mRNA and protein) in a time-dependent and dose-dependent manner. VEGF-B, VEGF-C and VEGF-D were expressed constitutively, and no further induction was observed in stimulated PCHLFs. LTA did not increase the expression levels of the VEGF family. TLR2 mRNA and protein were increased only when PCHLFs were stimulated with PGN. Treatment with an anti-TLR2 antibody blocked the interaction of PGN with the receptor, inhibiting VEGF-A over-expression; the presence of anti-TLR2 antibodies did not affect the over-production of VEGF-A after MDP treatment. PCHLFs stimulated with PGN and MDP, but not with LTA, activated NFκB. MDP stimulated the production of NOD1 and NOD2 mRNAs in a time-dependent and dose-dependent manner, and NOD2 protein was only increased by MDP. Treatment of PCHLFs with AG1478 and PD98059 inhibitors prior to stimulation with MDP resulted in the inhibition of VEGF-A over-production, compared with PCHLFs stimulated with MDP alone. Taken together, these results suggest that limbal fibroblasts produce VEGF-A through PGN-TLR2-NFκB and MDP-NOD2-EGFR.
- Published
- 2012
14. ATP-citrate lyase activity and carotenoid production in batch cultures of Phaffia rhodozyma under nitrogen-limited and nonlimited conditions
- Author
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Luis B. Flores-Cotera, Zoila R. Flores-Bustamante, Cipriano Chávez-Cabrera, Rodolfo Marsch, Juan C. Cancino-Diaz, Sergio Sánchez, and María del Carmen Montes
- Subjects
chemistry.chemical_classification ,Fungal protein ,ATP citrate lyase ,Nitrogen ,Basidiomycota ,food and beverages ,Fatty acid ,ATP Citrate (pro-S)-Lyase ,General Medicine ,Biology ,musculoskeletal system ,Lyase ,Carotenoids ,Applied Microbiology and Biotechnology ,Yeast ,Fungal Proteins ,Quaternary Ammonium Compounds ,chemistry.chemical_compound ,chemistry ,Biochemistry ,Acetyl Coenzyme A ,Astaxanthin ,Carotenoid ,Biotechnology - Abstract
ATP-citrate lyase (ACL) is the key cytoplasmic enzyme which supplies acetyl-CoA for fatty acids in oleaginous yeast. Although it has been suggested that fatty acid and carotenoid biosynthesis may have a common source of acetyl-CoA in Phaffia rhodozyma, the source for carotenoids is currently unknown. The purpose of this work was to analyze the development of ACL activity during batch cultures of P. rhodozyma under ammonium-limited and nonammonium-limited conditions and study its possible relationship with carotenoid synthesis. Every experiment showed carotenoid accumulation linked to an increasing ACL activity. Moreover, the ACL activity increased with dissolved oxygen (DO), i.e., ACL responded to DO in a similar way as carotenoid synthesis. Additionally, in the ammonium-limited culture, ACL activity increased upon ammonium depletion. However, the contribution to carotenoid accumulation in that case was negligible. This suggests that P. rhodozyma has developed two components of ACL, each one responsive to a different environmental stimulus, i.e., DO and ammonium depletion. The role of each component is still unknown; however, considering that the former responds to DO and the known role of carotenoids as antioxidants, it may be a provider of acetyl-CoA for carotenoid synthesis.
- Published
- 2009
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