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This study explores the potential of zeolite as an amendment to mitigate ammonium inhibition in the anaerobic digestion of swine waste. Two 50 L reactors, one with and one without zeolite amendment were operated at an OLR of 3.0 g VS L-1d-1 for 130 days, and fed with swine waste from a full-scale pig farm. Under these conditions, zeolite doses of 4 g L-1 allowed total ammonia nitrogen (TAN) concentrations to be kept below 1000 mgNH3-N L-1. The zeolite-amended reactor not only showed an average increase of 8% in methane production under stable conditions but also exhibited 34% reduction in H2S concentrations in the biogas, compared to the reactor without zeolite. The community of archaea originating from the inoculum was conserved in the reactor with zeolite amendment, particularly the acetoclastic methanogens of the genus Methanosaeta. On the other hand, in the reactor without zeolite addition, the microbial community went from being dominated by the acetoclastic methanogen Methanosaeta to having a high relative abundance of hydrogenotrophic methanogens. The zeolite addition also favoured the reactor stability, prevented foaming, and produced an enriched natural zeolite with N, P and K. However, additional studies on the potential of enriched zeolite as a fertilizer are required, which could make the use of zeolite in Anaerobic Digestion of swine waste not only energetically favourable but also economically feasible.
Assuntos
Zeolitas , Animais , Suínos , Anaerobiose , Reatores Biológicos , Equador , MetanoRESUMO
Anaerobic digestion (AD) of swine waste allows obtaining renewable energy, biofertilizer and the reduction of environmental impacts. However, the low C:N ratio of pig manure generates high concentrations of ammonia nitrogen in the digestion process, reducing methane production. Zeolite is an effective ammonia adsorbent; thus, in this research the ammonia adsorption capacity of natural Ecuadorian zeolite was studied under different operating conditions. Subsequently, its effect on methane production from swine waste was evaluated using three doses of zeolite, 1.0, 4.0 and 8.0 g, in 1 L batch bioreactors. The results showed that the Ecuadorian natural zeolite has an adsorption capacity of around 19 mgNH3-N gZ-1 when using ammonium chloride solution and, an adsorption capacity between 37 and 65 mgNH3-N gZ-1 using swine waste. On the other hand, the addition of zeolite had a significant effect on methane production (p < 0.01). The zeolite doses that provided the highest methane production were 4.0 and 8.0 g L-1, which led to values of 0.375 and 0.365 Nm3CH4 kgVS-1, compared to the values of 0.350 and 0.343 Nm3CH4 kgVS-1 that were obtained for the treatments without addition of zeolite and using a dose of 1.0 g L-1, respectively. Addition of natural Ecuadorian zeolite meant not only a significant increase on methane production in the AD of swine waste, but also a better quality of the biogas with higher percentages of methane and lower concentrations of H2S.
Assuntos
Zeolitas , Animais , Suínos , Anaerobiose , Amônia , Equador , Reatores Biológicos , Biocombustíveis , Esterco , MetanoRESUMO
The placenta can be affected by environmental factors, such as exposure to cigarette smoke. This exposure in the fetal context is considered a risk factor for the development of short-term postnatal diseases, such as asthma. Asthma is an inflammatory disease characterized by predominant acquisition of CD4 T lymphocytes (TLs) of the Th2 type. Transcription factors such as GATA binding protein 3 (GATA3) and STAT6 actively participate in the differentiation of virgin TLs towards the Th2 profile, while transcription factors such as STAT1, T-Box transcription factor 21 (T-BET), RUNX1 and RUNX3 participate in their differentiation towards the Th1 profile. The objective of the current study was to evaluate the impact of exposure to cigarette smoke on the gene expression of STAT1, T-BET, GATA3, IL-4, RUNX1 and RUNX3 during the gestation period, and to determine whether the expression levels of these genes are associated with changes in global methylation. STAT1, GATA3, RUNX1 and RUNX3 protein and mRNA expression levels in the placental tissue of women smokers and non-smoking women were determined via immunohistochemistry and quantitative PCR (qPCR) respectively. Additionally, T-BET and IL-4 mRNA expression levels were determined by qPCR. On the other hand, global methylation was determined via ELISA. In the present study, significant increases were observed in RUNX1 transcription factor expression in placentas from women smokers when compared with placentas of non-smoking women. Similarly, significant increases in the expression of GATA3, IL-4 and RUNX3 mRNA were observed. The changes in gene expression were not associated with changes in the global methylation levels. Finally, a higher frequency of low-birth-weight infants were identified in cases of exposure to cigarette smoke during pregnancy when compared with infants not exposed to cigarette smoke during pregnancy. Thus, the data of the present study contributed to the understanding of the genetic and clinical impacts of exposure to cigarette smoke during pregnancy and its importance in maternal and fetal health.
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OBJECTIVE: Antibodies against carbamylated proteins/peptide (CarP) have been associated with severity in rheumatoid arthritis (RA) patients. However, their role in risk groups, specific targets and relation with periodontal disease (PD) is uncertain yet. The aim of this study was evaluated the association between the levels of anti-CarP with clinical manifestation, human leukocyte antigen (HLA) alleles, periodontal activity markers, PD diagnosis, PD severity, and presence of Porphyromonas gingivalis (P gingivalis) in relatives of patients with RA. METHODS: One hundred and twenty-four individuals with a family history of RA in first-degree relatives (FDR) and 124 healthy individuals gender- and age-matched, RA activity was assessed. Antibodies against carbamylated protein anti-FCS-Carp and 2 carbamylated peptides of fibrinogen were selected (anti-Ca-Fib2, anti-Ca-Fib3). RESULTS: Anti-FCS-Carp-positive, anti-Ca-Fib2 and anti-Ca-Fib3 were more frequent in FDR than controls (25.0% vs 14.5%, 34.7% vs 15.3% and 33.1% vs 11.3%, respectively). Anti-FCS-CarP were associated with the HLA-DRB1-SE* 1402 allele (P = .035) and highly sensitive C-reactive protein levels (P = .016), the anti-Ca-Fib2 antibodies were associated with the HLA-DRB1-SE* 1501 allele (P = .03), with non-SE* 0901 allele (P = .01), the anti-Ca-Fib3 was associated with positive rheumatoid factor (P = .0012). The FDR condition was associated with the presence of anti-Ca-Fib3 (odds ratio [OR] =4.7; 95% CI = 1.8-11.7; P = .001) and painful joints (OR = 2.2; 95% CI = 1.01-4.68; P = .045); we also detected an important trend toward the presence of P gingivalis (OR = 1.9; 95% CI = 0.9-3.7; P = .062). CONCLUSION: The presence of anti-FCS-Carp, anti-Ca-Fib3 and anti-Ca-Fib2 antibodies may have a role for these antibodies as early biomarkers in the development of RA, probably including additional mechanisms related with other non-SE alleles; the anti-peptide antibodies proposed in the present study may represent a simpler way to identify antibodies directed to a specific target.
Assuntos
Artrite Reumatoide/imunologia , Autoanticorpos/imunologia , Carbamatos/imunologia , Adulto , Artrite Reumatoide/diagnóstico , Artrite Reumatoide/metabolismo , Biomarcadores/metabolismo , Carbamatos/metabolismo , Estudos Transversais , Feminino , Humanos , Inflamação/imunologia , Inflamação/metabolismo , Masculino , Carbamilação de ProteínasRESUMO
The genetic basis for sporadic immunodeficiency in patients with 22q11.2 distal deletion syndrome is unknown. We report an adult with a type 1 (D-F) 22q11.2 distal deletion syndrome and recurrent severe infections due to herpes zoster virus, presenting mild T cell lymphopenia and diminished frequency of naive CD4+ T cells, but increased frequencies of central, effector, and terminally differentiated memory T cells. Antigen-specific CD4+ and CD8+ T cells to influenza, rotavirus, and SEB were conserved in the patient, but responses to tetanus toxoid were temporarily undetectable. Exomic sequencing identified the c.20_22dupCGG (NM_002745.4) variant in the remaining MAPK1 gene of the patient, which adds 1 alanine to the polyalanine amino-terminal tract of the protein (p.Ala7dup). The mother, unlike the father, was heterozygote for the variant. Western blot analysis with the patient's activated PBMCs showed a 91% reduction in the MAPK1 protein. Further studies will be necessary to determine whether or not the variant present in the remaining MAPK1 gene of the patient is pathogenic.
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Transforming growth factor ß (TGF-ß) has been shown to play a role in immunity against different pathogens in vitro and against parasites in vivo However, its role in viral infections in vivo is incompletely understood. Using a neonatal mouse model of heterologous rhesus rotavirus (RV) vaccination, we show that the vaccine induced rotavirus-specific CD4 T cells, the majority of which lacked expression of KLRG1 or CD127, and a few regulatory rotavirus-specific CD4 T cells that expressed surface latency-associated peptide (LAP)-TGF-ß. In these mice, inhibiting TGF-ß, with both a neutralizing antibody and an inhibitor of TGF-ß receptor signaling (activin receptor-like kinase 5 inhibitor [ALK5i]), did not change the development or intensity of the mild diarrhea induced by the vaccine, the rotavirus-specific T cell response, or protection against a subsequent challenge with a murine EC-rotavirus. However, mice treated with anti-LAP antibodies had improved protection after a homologous EC-rotavirus challenge, compared with control rhesus rotavirus-immunized mice. Thus, oral vaccination with a heterologous rotavirus stimulates regulatory RV-specific CD4 LAP-positive (LAP+) T cells, and depletion of LAP+ cells increases vaccine-induced protection.IMPORTANCE Despite the introduction of several live attenuated animal and human rotaviruses as efficient oral vaccines, rotaviruses continue to be the leading etiological agent for diarrhea mortality among children under 5 years of age worldwide. Improvement of these vaccines has been partially delayed because immunity to rotaviruses is incompletely understood. In the intestine (where rotavirus replicates), regulatory T cells that express latency-associated peptide (LAP) play a prominent role, which has been explored for many diseases but not specifically for infectious agents. In this paper, we show that neonatal mice given a live oral rotavirus vaccine develop rotavirus-specific LAP+ T cells and that depletion of these cells improves the efficiency of the vaccine. These findings may prove useful for the design of strategies to improve rotavirus vaccines.
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Linfócitos T CD4-Positivos/imunologia , Infecções por Rotavirus/prevenção & controle , Vacinas contra Rotavirus/imunologia , Subpopulações de Linfócitos T/imunologia , Fator de Crescimento Transformador beta/análise , Administração Oral , Animais , Animais Recém-Nascidos , Linfócitos T CD4-Positivos/química , Diarreia/prevenção & controle , Modelos Animais de Doenças , Imunidade Heteróloga , Camundongos , Vacinas contra Rotavirus/administração & dosagem , Subpopulações de Linfócitos T/química , Resultado do TratamentoRESUMO
INTRODUCTION: Vitamin D3 (VD3) has been described as a modulator of immune system cells, including dendritic cells (DC). Previous studies have shown its importance in in vitro generation of tolerogenic DC, which have a similar function and phenotype to that of CD141 dermal DCs that produce IL-10 and induce (LTreg) CD4+ T regulator cells. OBJECTIVE: This paper presents a study that compares the phenotype and cytokines produced by DC generated in presence and absence of VD3, which were matured with lipopolysaccharide (LPS), and their ability to induce LTreg from naïve allogeneic CD4+ T cells. MATERIALS AND METHODS: In order to compare them, peripheral blood mononuclear cells were isolated to select monocytes CD14+ T cells and differentiate them in vitro from DC in the presence and absence of VD3, and to mature them with LPS. Phenotype and cytokine levels were also analyzed in the culture supernatants. Dendritic cells were then co-cultured with naïve allogeneic CD4+ T cells and the frequencies of LTreg were determined (naïve-activated). RESULTS: The results showed that unstimulated DC generated with VD3 kept the CD14. When activated with LPS, they expressed lower levels of C83, CD83 and CD86; HLA-DR; higher amounts of IL-1ß, IL-8, IL-10, and tended to lessen IL-6, IL-12p70 and TGF-ß1, compared to DCs not treated with VD3. The frequency of naïve LTreg was similar, although immature DC generated with VD3 tended to induce activated LTregs. CONCLUSION: Based on these results, it is possible to conclude that DCs generated with VD3 and treated with LPS presented a 'semi-mature' phenotype, and were able to secrete pro-inflammatory and anti-inflammatory cytokines. Besides, they did not increase their capacity to promote the polarization of naïve allogenic CD4+ T cells towards LTregs.
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Colecalciferol/farmacologia , Citocinas/biossíntese , Citocinas/imunologia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Interleucina-10/metabolismo , Interleucina-8/metabolismo , Leucócitos Mononucleares/imunologia , Lipopolissacarídeos/química , Monócitos/efeitos dos fármacos , Monócitos/imunologia , Linfócitos T Reguladores/imunologia , Células Cultivadas , Colecalciferol/metabolismo , Células Dendríticas/citologia , Humanos , Interleucina-10/imunologia , Interleucina-10/fisiologia , Interleucina-8/imunologia , Interleucina-8/fisiologia , Leucócitos Mononucleares/química , Leucócitos Mononucleares/fisiologia , Lipopolissacarídeos/metabolismo , Linfócitos T Reguladores/fisiologiaRESUMO
The response of antibody-secreting cells (ASC) induced by dengue has only recently started to be characterized. We propose that young age and previous infections could be simple factors that affect this response. Here, we evaluated the primary and secondary responses of circulating ASC in infants (6-12 months old) and children (1-14 years old) infected with dengue showing different degrees of clinical severity. The ASC response was delayed and of lower magnitude in infants, compared with older children. In primary infection (PI), the total and envelope (E) protein-specific IgM ASC were dominant in infants but not in children, and a negative correlation was found between age and the number of IgM ASC (rho = -0.59, P = 0.03). However, infants with plasma dengue-specific IgG detectable in the acute phase developed an intense ASC response largely dominated by IgG and comparable to that of children with secondary infection (SI). IgM and IgG produced by ASC circulating in PI or SI were highly cross-reactive among the four serotypes. Dengue infection caused the disturbance of B cell subsets, particularly a decrease in the relative frequency of naïve B cells. Higher frequencies of total and E protein-specific IgM ASC in the infants and IgG in the children were associated with clinically severe forms of infection. Therefore, the ASC response induced by dengue is highly influenced by the age at which infection occurs and previous immune status, and its magnitude is a relevant element in the clinical outcome. These results are important in the search for correlates of protection and for determining the ideal age for vaccinating against dengue.
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Anticorpos Antivirais/imunologia , Células Produtoras de Anticorpos/imunologia , Vírus da Dengue/imunologia , Dengue/imunologia , Proteínas do Envelope Viral/imunologia , Adolescente , Fatores Etários , Anticorpos Antivirais/sangue , Células Produtoras de Anticorpos/virologia , Subpopulações de Linfócitos B/imunologia , Subpopulações de Linfócitos B/virologia , Criança , Pré-Escolar , Reações Cruzadas/imunologia , Dengue/sangue , Dengue/virologia , Vírus da Dengue/genética , Vírus da Dengue/fisiologia , ELISPOT , Feminino , Interações Hospedeiro-Patógeno/imunologia , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoglobulina M/sangue , Imunoglobulina M/imunologia , Lactente , Masculino , SorogrupoRESUMO
Introducción. La vitamina D3 actúa como modulador de algunas células del sistema inmunitario, incluidas las células dendríticas. Varios estudios han reportado su importancia en la generación in vitro de células dendríticas tolerogénicas, similares en cuanto a fenotipo y función a las células dendríticas dérmicas CD141 productoras de IL-10 e inductoras de linfocitos T reguladores CD4+. Objetivo. Se compararon el fenotipo y las citocinas producidas por las células dendríticas generadas en ausencia o en presencia de la vitamina D3, y maduradas con lipopolisacáridos, así como su habilidad de inducir linfocitos T reguladores a partir de linfocitos T CD4+ vírgenes alogénicos. Materiales y métodos. Se aislaron células mononucleares de sangre periférica para seleccionar monocitos CD14+ y diferenciarlos in vitro de las células dendríticas en presencia o en ausencia de vitamina D3, y madurarlas con lipopolisacáridos. Se analizaron el fenotipo y los niveles de las citocinas en los sobrenadantes de cultivo. Se hizo un cocultivo de las células dendríticas con linfocitos T CD4+ vírgenes alogénicos y se determinaron las frecuencias de LTreg (vírgenes activados). Resultados. Las células dendríticas no estimuladas generadas con la vitamina D3 conservaron el CD14. Al activarlas con lipopolisacáridos, expresaron bajos niveles de C83, CD83 y CD86, HLA-DR, cantidades elevadas de IL-1ß, IL-8 e IL-10, y una tendencia a la disminución de IL-6, IL-12p70 y TGF-ß1 con respecto a las que no habían sido tratadas con la vitamina. La frecuencia de los LTreg vírgenes fue similar, aunque se observó una tendencia de las células dendríticas inmaduras generadas con la vitamina a inducir LTreg activados. Conclusión. Las células dendríticas generadas con vitamina D3 y tratadas con lipopolisacáridos presentaron un fenotipo 'semimaduro', así como la capacidad de secretar citocinas antiinflamatorias y citocinas promotoras de la reacción inflamatoria. Además, no se aumentó su capacidad de promover la polarización de LTCD4+ vírgenes alogénicos hacia LTreg.
Introduction: Vitamin D3 (VD3) has been described as a modulator of immune system cells, including dendritic cells (DC). Previous studies have shown its importance in in vitro generation of tolerogenic DC, which have a similar function and phenotype to that of CD141 dermal DCs that produce IL-10 and induce (LTreg) CD4+ T regulator cells. Objective: This paper presents a study that compares the phenotype and cytokines produced by DC generated in presence and absence of VD3, which were matured with lipopolysaccharide (LPS), and their ability to induce LTreg from naïve allogeneic CD4+ T cells. Materials and methods: In order to compare them, peripheral blood mononuclear cells were isolated to select monocytes CD14+ T cells and differentiate them in vitro from DC in the presence and absence of VD3, and to mature them with LPS. Phenotype and cytokine levels were also analyzed in the culture supernatants. Dendritic cells were then co-cultured with naïve allogeneic CD4+ T cells and the frequencies of LTreg were determined (naïve-activated). Results: The results showed that unstimulated DC generated with VD3 kept the CD14. When activated with LPS, they expressed lower levels of C83, CD83 and CD86; HLA-DR; higher amounts of IL-1ß, IL-8, IL-10, and tended to lessen IL-6, IL-12p70 and TGF-ß1, compared to DCs not treated with VD3. The frequency of naïve LTreg was similar, although immature DC generated with VD3 tended to induce activated LTregs. Conclusion: Based on these results, it is possible to conclude that DCs generated with VD3 and treated with LPS presented a 'semi-mature' phenotype, and were able to secrete pro-inflammatory and anti-inflammatory cytokines. Besides, they did not increase their capacity to promote the polarization of naïve allogenic CD4+ T cells towards LTregs.