Effect of Ecuadorian natural zeolite on the performance of anaerobic digestion of swine waste in semicontinuous regime.

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.

Natural Ecuadorian zeolite: An effective ammonia adsorbent to enhance methane production from swine waste.

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.

RUNX1 gene expression changes in the placentas of women smokers.

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.

Anti-carbamylated protein and peptide antibodies as potential inflammatory joint biomarkers in the relatives of rheumatoid arthritis patients.

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.

Immunodeficiency in a Patient with 22q11.2 Distal Deletion Syndrome and a p.Ala7dup Variant in the MAPK1 Gene.

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.

LAP+ Cells Modulate Protection Induced by Oral Vaccination with Rhesus Rotavirus in a Neonatal Mouse Model.

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.

Dendritic cells generated in the presence of vitamin D3 and stimulated with lipopolysaccharide secrete IL-8, IL-1ß, IL-10 and induce relatively low levels of CD4+CD25hiFoxp3+ T cells.

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.

Total and Envelope Protein-Specific Antibody-Secreting Cell Response in Pediatric Dengue Is Highly Modulated by Age and Subsequent Infections.

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.

Las células dendríticas generadas en presencia de vitamina D3 y activadas con lipopolisacáridos incrementan la producción de IL-1ß, IL-8 e IL-10 y disminuyen su capacidad de inducir LT CD4+CD25hiFoxp3+ / Dendritic cells generated in the presence of vitamin D3 and stimulated with lipopolysaccharide secrete IL-8, IL-1ß, IL-10 and induce relatively low levels of CD4+CD25hiFoxp3+ T cells

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.

La sertralina incluida en la ß-ciclodextrina no induce un perfil tolerogénico en células dendríticas derivadas de monocitos / Title: Sertraline Included in ß-ciclodextrin does not Induce a Tolero- genic Profile in Monocyted-Derived Dendritic Cells

Objetivo: Evaluar el efecto inmunomodulador de la sertralina (SRT) y la sertralinaincluida en la ß-ciclodextrina (LF) en células dendríticas humanas (CD) generadas invitro a partir de monocitos sobre marcadores de fenotipo y la producción de citocinas.Materiales y métodos: Se aislaron monocitos CD14+ de células mononucleares desangre periférica y se diferenciaron a CD; posteriormente, se pretrataron con SRT oLF durante una hora. Finalmente, las CD se maduraron con lipopolisacárido (LPS)durante 24 horas y se analizó el fenotipo de las CD y las concentraciones de citocinasen los sobrenadantes de cultivo. Resultados: No se observaron cambios al compararel fenotipo de las CD maduradas con LPS en ausencia o presencia de la SRT. Asímismo, no hubo variación en cuanto a la producción de citocinas. Conclusión: LaSRT incluida o no en la ß-ciclodextrina no afecta el fenotipo y la secreción de las CDtratadas con LPS.

To evaluate the immunomodulatory effect of sertraline (SRT) and sertraline inclu - ded in ß-cyclodextrin (LF) in human dendritic cells (DCs) generated in vitro from monocytes on phenotype markers and cytokine produc - tion. Materials and Methods: CD14 + mono - cytes from peripheral blood mononuclear cells were isolated and differentiated to DCs. DCs were subsequently pretreated with LF or SRT for one hour. Finally DCs were matured with lipopolysaccharide (LPS) for 24 hours and the CDs phenotype and the levels of cytokines in the culture supernatants were analyzed. Results: No change on the phenotype of the DCs matured with LPS in the absence or presence of the SRT was observed. Likewise, there was no variation in cytokine production. Conclusion : SRT or not including ß-cyclodextrin does not affect the phenotype and secretion of LPS-treated DCs.

Caco-2 cells infected with rotavirus release extracellular vesicles that express markers of apoptotic bodies and exosomes.

Previously, we showed that infecting human intestinal epithelial cells (Caco-2) with rotavirus (RV) increases the release of extracellular vesicles (EVs) with an immunomodulatory function that, upon concentration at 100,000×g, present buoyant densities on a sucrose gradient of between 1.10 to 1.18 g/ml (characteristic of exosomes) and higher than 1.24 g/ml (proposed for apoptotic bodies). The effect of cellular death induced by RV on the composition of these EV is unknown. Here, we evaluated exosome (CD63, Hsc70, and AChE) and apoptotic body (histone H3) markers in EVs isolated by differential centrifugation (4000×g, 10,000×g, and 100,000×g) or filtration/ultracentrifugation (100,000×g) protocols. When we infected cells in the presence of caspase inhibitors, Hsc70 and AChE diminished in EVs obtained at 100,000×g, but not in EVs obtained at 4000×g or 10,000×g. In addition, caspase inhibitors decreased CD63 and AChE in vesicles with low and high buoyant densities. Without caspase inhibitors, RV infection increased exosome markers in all of the EVs obtained by differential centrifugation. However, CD63 preferentially localized in the 100,000×g fraction and H3 only increased in EVs concentrated at 100,000×g and with high buoyant densities on a sucrose gradient. Thus, RV infection increases the release of EVs that, upon concentration at 100,000×g, are composed by exosomes and apoptotic bodies, which can partially be separated using sucrose gradients.

Circulating rotavirus-specific T cells have a poor functional profile.

Frequencies of circulating T cells producing IFN-γ, TNF-α, and IL-2, and percentages of T cells proliferating after stimulation with rotavirus (RV), tetanus toxoid, and influenza were evaluated in PBMC derived from healthy adults and children. In addition, the potential anergic state of RV-specific T cells was analyzed by stimulation of PBMC with RV antigen in the presence of three anergy inhibitors (rIL-2, rIL-12, or DGKα-i). The quality and magnitude of RV-T cell responses were significantly lower than those of tetanus toxoid and influenza antigens. RV-CD4 T cell response was enriched in monofunctional IFN-γ(+) cells, while influenza-CD4 and tetanus toxoid-CD4 T cell responses were enriched in multifunctional T cells. Moreover, rIL-2--unlike rIL-12 or DGKα-i--increased the frequencies of RV-CD4 TNF-α(+), CD4 IFN-γ(+), and CD8 IFN-γ(+) cells. Thus, circulating RV-T cells seem to have a relatively poor functional profile that may be partially reversed in vitro by the addition of rIL-2.

Production of volatile fatty acids from wastewater screenings using a leach-bed reactor.

Screenings recovered from the inlet works of wastewater treatment plants were digested without pre-treatment or dilution using a lab-scale, leach-bed reactor. Variations in recirculation ratio of the leachate of 4 and 8 l/lreactor/day and pH values of 5 and 6 were evaluated in order to determine the optimal operating conditions for maximum total volatile fatty acids (VFA) production. By increasing the recirculation ratio of the leachate from 4 to 8 l/lreactor/day it was possible to increase VFA production (11%) and soluble COD (17%) and thus generate up to 264 g VFA/kg-dry screenings. These VFA were predominantly acetic acid with some propionic and butyric acid. The optimum pH for VFA production was 6.0, when the methanogenic phase was inhibited. Below pH 5.0, acid-producing fermentation was inhibited and some alcohols were produced. Ammonia release during the hydrolysis of screenings provided adequate alkalinity; consequently, a digestion process without pH adjustment could be recommended. The leach-bed reactor was able to achieve rapid rates of screenings degradation with the production of valuable end-products that will reduce the carbon footprint associated with current screenings disposal techniques.

High plasma levels of soluble ST2 but not its ligand IL-33 is associated with severe forms of pediatric dengue.

Identification of early determinants of dengue disease progression, which could potentially enable individualized patient care are needed at present times. Soluble ST2 (sST2) has been recently reported to be elevated in the serum of children older than 2 years old and adults with dengue infection and it was correlated with secondary infections as well as with severe presentations of the disease. The mechanism by which secreted ST2 is linked to severe dengue and plasma leakage remains unclear. One possibility is that IL-33 ligand may be elevated, contributing to membrane bound ST2 as part of the immune activation in dengue infection. We determined plasma levels of sST2 and the ligand IL-33 in 66 children with acute secondary dengue infections clinically classified using the guidelines of the World Health Organization, 2009. Dengue infection showed significant increases in cytokines IL-12p70, IL-10, IL-8, IL-6, IL-1ß and TNFα measured by flow cytometry based assay compared to uninfected individuals. In contrast, IL-33 levels remained unchanged between infected and uninfected individuals. The levels of sST2 positively correlated with values of IL-6 and IL-8 and inversely correlated with number of median value of platelet levels. In addition to circulating cytokine positive correlations we found that sST2 and isoenzyme creatine kinase-MB (CK-MB), a marker of myocardial muscle damage present in severe dengue cases were associated. Our pediatric study concluded that in dengue infections sST2 elevation does not involve concomitant changes of IL-33 ligand. We propose a study to assess its value as a predictor factor of disease severity.

Human myeloid dendritic cells treated with supernatants of rotavirus infected Caco-2 cells induce a poor Th1 response.

We have previously shown that human myeloid dendritic cells treated with purified rotavirus induce an allogenic Th1 response. To determine if rotavirus in the context of an intestinal microenvironment modulates the function of dendritic cells, we treated these cells with supernatants from non-infected or infected Caco-2 cells and studied their capacity to promote Th1 or Th2 responses. Dendritic cells treated with supernatants from rotavirus-infected Caco-2 cells promoted a significantly lower Th1 response, in comparison with those treated with purified rotavirus. We wanted to establish if TGF-ß1, induced, or TSLP, not induced, during rotavirus infection, could mediate this effect. Neutralization of TGF-ß but not TSLP in the supernatant prior to treatment of dendritic cells increased their capacity to promote a Th1 response. The results suggest that the TGF-ß1 induced by rotavirus could be an immune evasion mechanism, and may partially explain the poor rotavirus-specific T cell response we have previously evidenced.

Membrane vesicles released by intestinal epithelial cells infected with rotavirus inhibit T-cell function.

Rotavirus (RV) predominantly replicates in intestinal epithelial cells (IEC), and "danger signals" released by these cells may modulate viral immunity. We have recently shown that human model IEC (Caco-2 cells) infected with rhesus-RV release a non-inflammatory group of immunomodulators that includes heat shock proteins (HSPs) and TGF-ß1. Here we show that both proteins are released in part in association with membrane vesicles (MV) obtained from filtrated Caco-2 supernatants concentrated by ultracentrifugation. These MV express markers of exosomes (CD63 and others), but not of the endoplasmic reticulum (ER) or nuclei. Larger quantities of proteins associated with MV were released by RV-infected cells than by non-infected cells. VP6 co-immunoprecipitated with CD63 present in these MV, and VP6 co-localized with CD63 in RV-infected cells, suggesting that this viral protein is associated with the MV, and that this association occurs intracellularly. CD63 present in MV preparations from stool samples from 36 children with gastroenteritis due or not due to RV were analyzed. VP6 co-immunoprecipitated with CD63 in 3/8 stool samples from RV-infected children, suggesting that these MV are released by RV-infected cells in vivo. Moreover, fractions that contained MV from RV-infected cells induced death and inhibited proliferation of CD4(+) T cells to a greater extent than fractions from non-infected cells. These effects were in part due to TGF-ß, because they were reversed by treatment of the T cells with the TGF-ß-receptor inhibitor ALK5i. MV from RV-infected and non-infected cells were heterogeneous, with morphologies and typical flotation densities described for exosomes (between 1.10 and 1.18 g/mL), and denser vesicles (>1.24 g/mL). Both types of MV from RV-infected cells were more efficient at inhibiting T-cell function than were those from non-infected cells. We propose that RV infection of IEC releases MV that modulate viral immunity.

Análisis de la justicia restaurativa para atender casos de violencia intrafamiliar en el Centro de Atención Integral a Víctimas de Violencia Intrafamiliar (CAVIF) de la Fiscalía General de la Nación, Colombia / Analysis of restorative justice attending cases of Intrafamiliar Violence in the Integral Centre for Victims of Domestic Violence (CAVIF) - Fiscalía General de la Nación, Colombia

La presente investigación hace un análisis de la Justicia Restaurativa orientado a la atención de casos de violencia intrafamiliar en el Centro de Atención Integral a Víctimas de Violencia Intrafamiliar (CAVIF) de la Fiscalía General de la Nación. Este análisis es el producto de un trabajo conceptual y un ejercicio metodológico aplicado a un proceso de recolección, sistematización y análisis de la información, basado en un diseño de investigación exploratoria. Para el estudio se tuvo una muestra intencional, integrada por 20 usuarias y 18 profesionales del CAVIF, a quienes se les administraron entrevistas a profundidad, diseñadas a partir de categorías previamente definidas. De esta manera, se describe una serie de necesidades del contexto institucional, profesional y de los propios usuarios y funcionarios del sistema, las cuales permiten comprender las condiciones necesarias, para proponer unos lineamientos generales para la formulación de un programa de justicia restaurativa, acorde con la ley vigente en Colombia. Los resultados del estudio permiten señalar la necesidad de llevar a cabo procesos de capacitación en justicia restaurativa con los funcionarios responsables de dar trámite a los conflictos de violencia intrafamiliar en el CAVIF. Como aporte final se proponen criterios para ejecutar un programa de justicia restaurativa.

The actual investigation makes an emphasis about restorative justice, its oriented to catch the attention for those cases of inner familiar violence, cases that are being taking care of the integral centre attention for victims of domestic violence (CAVIF). Existence of the following analysis appears on the basis of conceptual work and methodological exercise, all applied from a process of collecting, systematizing and analyzing information based on a design of exploratory investigation. Study had an intentional sample, 20 female users and 18 professionals from (CAVIF) proceeded with a series of profound interviews designed based on categories previously defined. This way it's described all kind of context needs, institutional, professional and own users and functionaries of the system in order to comprehend the necessary conditions, proposing general alignments for the formulation of a restorative justice on accordance of actual by law in Colombia. Results of the study are permitting point to the necessity of carry on different process of capacitating the restorative justice functionaries, responsible for allowing the necessary steps to the domestic violence conflicts in the (CAVIF). As a last it is a criteria proposed in order to carry on a justice restorative program.

Immunomodulators released during rotavirus infection of polarized caco-2 cells.

Rotavirus preferentially replicates in enterocytes and "danger signals" released by these cells are likely to modulate viral immunity. As a model of these events, we studied selected immunomodulators released during rotavirus infection of polarized Caco-2 cells grown in transwell cultures (TW). At early time points post-infection the virus was detected mainly in the apical side of the TWs, but this tendency was progressively lost concomitantly with disruption of the cell monolayer and cell death. Rotavirus-infected cells released IL-8, PGE(2), small quantities of TGF-beta1, and the constitutive and inducible heat shock proteins HSC70 and HSP70, but not IL-1beta, IL-6, IL-10, IL-12p70, or TNF-alpha. This set of immunomodulators is known to induce a non-inflammatory (non-Th-1) immune response, and may be determining, in part, the relatively low T-cell immune response observed in blood samples after RV infection.

Paraganglioma maligno retroperitoneal en el adulto / Malignant retroperitoneal paraganglioma in adults

Se reporta el caso de un paraganglioma retroperitoneal maligno de tipo no familiar, debido a su baja incidencia, y, en este caso en particular, por su comportamiento biológico agresivo con extenso compromiso abdominal y metastásico. Debido a que el diagnóstico histopatológico de malignidad es difícil, las características de infiltración local o las metástasis a distancia demostradas por imágenes —especialmente por CT y RM— son útiles para su diagnóstico. La gammagrafía con MIBG y el PET con nuevos radiotrazadores no sólo detectan correctamente la enfermedad metastásica, sino que, también, identifican candidatos para tratamiento con radiofármacos. Debe considerarse el diagnóstico de paraganglioma ante la presencia de una masa hipervascularizada ubicada en áreas anatómicas donde existe paraganglia, en un paciente con catecolaminas elevadas y glándulas adrenales normales.