Bifidobacterium longum and Chlorella sorokiniana Combination Modulates IFN-γ, IL-10, and SOCS3 in Rotavirus-Infected Cells.

Rotavirus is the main cause of acute diarrhea in children up to five years of age. In this regard, probiotics are commonly used to treat or prevent gastroenteritis including viral infections. The anti-rotavirus effect of Bifidobacterium longum and Chlorella sorokiniana, by reducing viral infectivity and improving IFN-type I response, has been previously reported. The present study aimed to study the effect of B. longum and/or C. sorokiniana on modulating the antiviral cellular immune response mediated by IFN-γ, IL-10, SOCS3, STAT1, and STAT2 genes in rotavirus-infected cells. To determine the mRNA relative expression of these genes, HT-29 cells were treated with B. longum and C. sorokiniana alone or in combination, followed by rotavirus infection. In addition, infected cells were treated with B. longum and/or C. sorokiniana. Cellular RNA was purified, used for cDNA synthesis, and amplified by qPCR. Our results demonstrated that the combination of B. longum and C. sorokiniana stimulates the antiviral cellular immune response by upregulating IFN-γ and may block pro-inflammatory cytokines by upregulating IL-10 and SOCS3. The results of our study indicated that B. longum, C. sorokiniana, or their combination improve antiviral cellular immune response and might modulate pro-inflammatory responses.

Prevalence of Adverse Reactions to Gluten and People Going on a Gluten-Free Diet: A Survey Study Conducted in Brazil.

BACKGROUND: The prevalence of gluten-related disorders (GRD) and adherence to a gluten-free diet (GFD) remains unknown in Brazilian population and there is no published information on the scientific literature about the proportion of Brazilians that were diagnosed with a gluten-related disorder. Thus, the aim of this work was to estimate the prevalence of GRDs and adherence to a GFD by self-report in adult Brazilian population. MATERIALS AND METHODS: A questionnaire-based cross-sectional study was conducted in two Brazilian cities. RESULTS: The response rate was 93.2% (1630/1749). The self-reported prevalence rates were (95% CI): adverse reactions to gluten 10.06% (8.64-11.62); gluten sensitivity 2.33% (1.65-3.18); physician-diagnosed celiac disease 0.3% (0.09-0.71); non-celiac gluten sensitivity 1.71% (1.14-2.47); wheat allergy 0.79% (0.42-1.36); adherence to gluten-free diet 7.48% (6.25-8.87); gluten avoiders 15.21% (13.5-17.05). Among those who were following a GFD (n = 122), 65.6% (n = 80) of them reported that they did not develop symptoms after wheat/gluten ingestion and 50% (n = 61) were following the diet without medical/dietitian advice. The main motivation for following a GFD in the self-reported and non-self-reported gluten sensitivity groups were the symptoms triggered after wheat/gluten ingestion (86.8%) and weight control (57.1%), respectively. CONCLUSIONS: Implementation of programs to increase awareness about GRDs among healthcare professionals and giving scientifically sound information to the general population about the risks and benefits for following a GFD are desirable actions in Brazil. The results also add to the growing body of evidence for highlighting the under-diagnosis of GRD and the trend for following a GFD in Latin America.

Modulation of rotavirus severe gastroenteritis by the combination of probiotics and prebiotics.

Annual mortality rates due to infectious diarrhea are about 2.2 million; children are the most vulnerable age group to severe gastroenteritis, representing group A rotaviruses as the main cause of disease. One of the main factors of rotavirus pathogenesis is the NSP4 protein, which has been characterized as a viral toxin involved in triggering several cellular responses leading to diarrhea. Furthermore, the rotavirus protein NSP1 has been associated with interferon production inhibition by inducing the degradation of interferon regulatory factors IRF3, IRF5, and IRF7. On the other hand, probiotics such as Bifidobacterium and Lactobacillus species in combination with prebiotics such as inulin, HMO, scGOS, lcFOS have been associated with improved generalized antiviral response and anti-rotavirus effect by the reduction of rotavirus infectivity and viral shedding, decreased expression of NSP4 and increased levels of specific anti-rotavirus IgAs. Moreover, these probiotics and prebiotics have been related to shorter duration and severity of rotavirus diarrhea, to the prevention of infection and reduced incidence of reinfections. In this review we will discuss in detail about the rotavirus pathogenesis and immunity, and how probiotics such as Lactobacillus and Bifidobacterium species in combination with prebiotics have been associated with the prevention or modulation of rotavirus severe gastroenteritis.

IL-10 and socs3 Are Predictive Biomarkers of Dengue Hemorrhagic Fever.

BACKGROUND: Cytokines play important roles in the physiopathology of dengue infection; therefore, the suppressors of cytokine signaling (socs) that control the type and timing of cytokine functions could be involved in the origin of immune alterations in dengue. OBJECTIVE: To explore the association of cytokine and socs levels with disease severity in dengue patients. METHODS: Blood samples of 48 patients with confirmed dengue infection were analyzed. Amounts of interleukins IL-2, IL-4, IL-6, and IL-10, interferon- (IFN-) γ, and tumor necrosis factor- (TNF-) α were quantified by flow cytometry, and the relative expression of socs1 and socs3 mRNA was quantified by real-time RT-PCR. RESULTS: Increased levels of IL-10 and socs3 and lower expression of socs1 were found in patients with dengue hemorrhagic fever (DHF) with respect to those with dengue fever (DF) (p < 0.05). Negative correlations were found between socs1 and both IL-10 and socs3 (p < 0.01). The cutoff values of socs3 (>199.8-fold), socs1 (<1.94-fold), and IL-10 (>134 pg/ml) have the highest sensitivity and specificity to discriminate between DF and DHF. CONCLUSION: Simultaneous changes in IL-10 and socs1/socs3 could be used as prognostic biomarkers of dengue severity.

Interferon lambda inhibits dengue virus replication in epithelial cells.

BACKGROUND: In viral disease, infection is controlled at the cellular level by type I interferon (IFN-I), but dengue virus (DENV) has the ability to inhibit this response. Type III interferon, also known as lambda IFN (IFN-III or IFN-λ), is a complementary pathway to the antiviral response by IFN-I. This work analyzed the IFN-λ (IFN-III) mediated antiviral response against DENV serotype 2 (DENV-2) infection. METHODS: Dengue fever patients were sampled to determine their IFN-λ levels by ELISA. To study the IFN-λ response during DENV infection we selected the epithelial cell line C33-A, and we demonstrated that it is permissive to DENV-2 infection. The effect of IFN-λ on virus replication was determined in these cells, in parallel to the expression of IFN-stimulated genes (ISGs), and Suppressor of Cytokine Signaling (SOCS), genes measured by RT-qPCR. RESULTS: We found increased (~1.8 times) serological IFN-λ in dengue fever patients compared to healthy blood donors. IFN-λ inhibited DENV-2 replication in a dose-dependent manner in vitro. The reduction of viral titer corresponded with increased ISG mRNA levels (MX1 and OAS1), with the highest inhibition occurring at ISG's peak expression. Presence of IFN-negative regulators, SOCS1 and SOCS3, during DENV-2 infection was associated with reduced IFN-λ1 expression. CONCLUSIONS: Evidence described here suggests that IFN-λ is a good candidate inhibitor of viral replication in dengue infection. Mechanisms for the cellular and organismal interplay between DENV and IFN- λ need to be further studied as they could provide insights into strategies to treat this disease. Furthermore, we report a novel epithelial model to study dengue infection in vitro.

Bifidobacterium longum and Chlorella sorokiniana Improve the IFN Type I-Mediated Antiviral Response in Rotavirus-Infected Cells.

Probiotics are effective to treat or prevent gastrointestinal infections, and microalgae have demonstrated important health-promoting effects and in some cases function as prebiotics. In this regard, the anti-rotavirus effect of Bifidobacterium longum and Chlorella sorokiniana by reducing viral infectivity is well known. However, their effect on immune response against rotavirus has not yet been investigated. Therefore, the aim of this study was to determine the role of Bifidobacterium longum and/or Chlorella sorokiniana in influencing an IFN type I-mediated antiviral response in rotavirus-infected cells. In pre-infection experiments, HT-29 cells were treated with B. longum and C. sorokiniana alone or in combination, followed by rotavirus infection, whereas in post-infection assays, HT-29 cells were treated after infection. The cells' mRNA was then purified to determine the relative expression level of IFN-α, IFN-ß, and precursors of interferons such as RIG-I, IRF-3, and IRF-5 by qPCR. We showed that combination of B. longum and C. sorokiniana significantly increased IFN-α levels in pre-infection and IFN-ß in post-infection assays, as compared with individual effects. Results indicate that B. longum, C. sorokiniana, or their combination improve cellular antiviral immune response.

A Food Matrix Triggers a Similar Allergic Immune Response in BALB/c Mice Sensitized with Native, Denatured, and Digested Ovalbumin.

The search for an animal model to evaluate the allergenic potential of processed food products is still ongoing. Both the sensitization to ovalbumin (OVA) in different structural states and the allergic response triggered after intragastric or food challenges were assessed. BALB/c mice were sensitized intraperitoneally to OVA (50 µg) in different structural states (native OVA, N-OVA; denatured OVA, D-OVA; formaldehyde- and lysine-treated OVA, FK-OVA; denatured OVA-FK, OVA-DFK; peptides from pepsin digestion, Pep-OVA). Anti-OVA-specific IgE responses were evaluated using ELISA. Anaphylactic signs and mMCP-1 serum levels were evaluated after intragastric (2.0 mg/OVA) and food (0.41 mg/OVA) challenges. IgE reactivities to N-OVA and D-OVA were similar among groups (p > 0.05). After the challenges, all OVA-sensitized mice developed mild to severe anaphylactic signs (p < 0.05 vs. control). Mice sensitized to N-OVA and D-OVA had the highest mMCP-1 serum levels after challenges (p < 0.05 vs. control). Allergic responses were similar despite the different OVA doses used for the challenges. The N-OVA-sensitized murine model of egg allergy proposed in the present study holds the potential for evaluating the impact of food matrix composition and processing on the threshold of egg-allergic responses.

Low Activation of CD8+ T Cells in response to Viral Peptides in Mexican Patients with Severe Dengue.

It is acknowledged that antiviral immune response contributes to dengue immunopathogenesis. To identify immunological markers that distinguish dengue fever (DF) and dengue hemorrhagic fever (DHF), 113 patients with confirmed dengue infection were analyzed at 6 or 7 days after fever onset. Peripheral blood mononuclear cells (PBMC) were isolated, lymphocyte subsets and activation biomarkers were identified by flow cytometry, and differentiation of T helper (Th) lymphocytes was achieved by the relative expression analysis of T-bet (Th1), GATA-3 (Th2), ROR-γ (Th17), and FOXP-3 (T regulatory) transcription factors quantified by real-time PCR. CD8+, CD40L+, and CD45+ cells show higher numbers in DF compared to DHF patients, whereas CD4+, CD19+, and CD25+ cells show higher numbers in DHF than DF patients. High expression of GATA-3 accompanied by low expression of T-bet indicates predominance of Th2 response. In addition, higher expression of FOXP-3 and reduced functional cytotoxic T cells (CD8+perforin+) were observed in DHF patients. In further experiments, PBMC were stimulated ex vivo with dengue virus E, NS3, NS4, and NS5 peptides, and proliferating T cell subsets were determined. Lower proliferative responses to NS3 and NS4 peptides and reduced CD8+ cytotoxic T cells were observed in DHF patients. Our results suggest that immune response to dengue is dysregulated with predominance of CD4+ T cells, low activation of Th1 cells, and downregulation of the antiviral cytotoxic activity during severe dengue, likely induced by regulatory T cells.

Prediction of ACE-I Inhibitory Peptides Derived from Chickpea (Cicer arietinum L.): In Silico Assessments Using Simulated Enzymatic Hydrolysis, Molecular Docking and ADMET Evaluation.

Chickpea (Cicer arietinum L.) peptides have shown in vitro potential to inhibit the angiotensin I-converting enzyme (ACE-I). However, the potential molecular interactions between chickpea peptides (CP) and ACE-I as well as their ADMET (absorption/distribution/metabolism/excretion/toxicity) characteristics remain unknown. Thus, our aim was to study the in silico interactions of CP with ACE-I and the CP ADMET characteristics. Legumin and provicilin sequences were submitted to in silico analysis to search for ACE-I inhibitory peptides. Simulated enzymatic hydrolysis was performed using the BIOPEP-UWM database, and the ACE-I inhibitory peptides generated (EC50 ≤ 200 µM) were selected to perform molecular docking and ADMET analysis. After hydrolysis, 59 out of 381 peptides with ACE-I inhibitory potential were released. Based on A and B parameters, the legumin peptides showed better ACE-I inhibitory potential than the provicilin ones. CP mainly interact with residues from pocket S1 (Ala354/Glu384) and S2 (His353/His513) through hydrogen bonds (distances < 3.0 Å) and hydrophobic interactions (binding energy from −5.7 to −9.2 kcal/mol). Through ADMET analysis, CP showed optimal values for inhibiting ACE-I in vivo. ACE-I inhibitory peptides from legumin and provicilin can bind strongly and tightly to the active site of ACE-I. Further studies to evaluate in vivo the antihypertensive effects of CP are warranted.

Assessment of the Route of Exposure to Ovalbumin and Cow's Milk Proteins on the Induction of IgE Responses in BALB/c Mice.

BALB/c mice can be orally sensitized to food proteins under acid suppressive medication, mimicking human exposure and triggering a human-like allergic immune response. However, the reproducibility of such an oral food allergy model remains questionable. Our aim was to evaluate the IgE responses triggered against ovalbumin (OVA) and cow's milk proteins (CMP) after intragastric (IG), either under gastric-acid suppression or not, or intraperitoneal (IP) sensitization in BALB/c mice. OVA (0.2 mg) and different concentrations of CMP were administered with/without the antacid sucralfate by the IG route. For IP sensitization, OVA or CMP (0.5 mg) were administered. ELISA was used to evaluate IgE responses. The IP sensitization protocols triggered more robust and consistent anti-OVA or anti-CMP IgE responses than the intragastric ones (with/without sucralfate) (p < 0.05). 2.7% (1/36), and 5.5% (3/54) of the mice that underwent the sucralfate-assisted IG protocol triggered IgE responses against OVA or CMP, respectively. All the mice were administered OVA or CMP via IP triggered detectable IgE responses. The IP sensitization model is more reliable than the IG one for evaluating the intrinsic sensitizing and/or allergenic potential of food proteins, even if IG immunizations are carried out under gastric-acid suppression.

Prevalence of Wheat/Gluten-Related Disorders and Gluten-Free Diet in Paraguay: An Online Survey-Based Study.

Gluten-related disorders (GRDs) are increasing around the world, but their magnitude and relevance remain unknown in most Latin American countries. Thus, an online survey was conducted to estimate the prevalence of GRDs as well as adherence to a gluten-free diet (GFD) in Paraguayan adult population. There were 1058 individuals surveyed using a validated questionnaire (response rate of 93.9%). The self-reported prevalence rates were as follows (95% CI): gluten sensitivity (GS), 10.30% (8.53-12.29); non-celiac GS (NCGS), 5.19% (3.94-6.71); physician-diagnosed celiac disease (PD-CD), 3.11% (2.15-4.35); wheat allergy (WA), 2.07% (1.30-3.13); and adherence to GFD, 15.69% (13.55-18.02). Excluding CD, more women than men met the criteria for GRDs, adverse food reactions, and GFD (p < 0.05). Eight respondents reported the coexistence of NCGS with PD-CD and/or WA. Most cases on a GFD indicated medical/dietitian advice for following the diet (68.07%). Non-self-reported GS individuals indicated weight control (46.4%) and the notion that the GFD is healthier (20.2%) as the main motivations for following the diet. GRDs are not uncommon in Paraguayan adult population. It seems that there is awareness about GRDs and the GFD, but training about the diagnosis of GRDs is desirable because of the informed overlapping diagnoses of CD or WA with NCGS. Future studies involving face-to-face interviews are necessary.

Th17 Lymphocytes in Children with Asthma: Do They Influence Control?

Background: Allergic asthma was considered as an inflammation mediated by specific CD4+ helper lymphocytes (Th2); however, this paradigm changed in 2005, when a third group of helper cells called Th17 cells were identified. Th17 lymphocytes are the main source of interleukin (IL)-17A-F, IL-21, and IL-22; however, their physiological role in children is unclear. This study aimed to determine the percentage of Th17 cells and IL-17A in pediatric patients diagnosed with asthma and to associate it with disease control using a validated questionnaire. Methods: This cross-sectional, prospective, comparative study included 92 asthma-diagnosed children 4-18 years of age. The Asthma Control Test was used as an assessment measure to classify patients as controlled (n = 30), partially controlled (n = 31), and uncontrolled (n = 31). Th17 cells and IL-17A were analyzed by flow cytometry. Patients receiving inhaled steroid therapy as monotherapy or associated with a long-acting bronchodilator were included. Results: The mean percentage of Th17 cells in the participants was 4.55 ± 7.34 (Controlled), 5.50 ± 8.09 (Partially Controlled), and 6.14 ± 7.11 (Uncontrolled). There was no significant difference between the 3 groups (P = 0.71). The mean percentage of IL-17A in all the participants was 9.84 ± 9.4 (Controlled), 10.10 ± 10.5 (Partially Controlled), and 11.42 ± 8.96 (Uncontrolled); no significant difference between the 3 groups (P = 0.79) was observed. Th17 lymphocyte levels were similar among the 3 groups and the same trend was observed with IL-17A. A significant correlation between Th17 or IL-17A and the degree of asthma control (Th17, P = 0.24; IL-17A, P = 0.23) was not found. Conclusions: The percentages of both Th17 lymphocytes and IL-17A found in children with asthma were not significantly different in the 3 groups, which suggests that they do not play an important role in asthma control. Our findings may contribute to the knowledge related to non-Th2 inflammation in children. Clinical-Trials.gov ID: 2015-2102-85.

Enhanced Viability and Anti-rotavirus Effect of Bifidobacterium longum and Lactobacillus plantarum in Combination With Chlorella sorokiniana in a Dairy Product.

Microalgae and probiotics such as Bifidobacterium and Lactobacillus genera are associated with human beneficial effects. The aim of this study was to evaluate the activity of Chlorella sorokiniana on Bifidobacterium longum and Lactobacillus plantarum viability in a dairy product (flan) and its microbial effect against rotavirus, which is one of the major diarrhea-causing pathogens worldwide. Microalge were isolated from a Mexican river and characterized by molecular tools. Their prebiotic activity was evaluated by determining Bifidobacterium longum and Lactobacillus plantarum shelf-life after incorporation in the food matrix. In addition, HT-29 cells were infected with rotavirus Wa and treated with 1 × 109 CFU/mL L. plantarum and B. longum metabolites alone or in combination with 1 × 109 cells/mL Chlorella sorokiniana; viral titers in probiotics- and/or microalgae-treated cells were evaluated for antiviral activity. Results indicated that C. sorokiniana not only significantly (p < 0.05) improved L. plantarum and B. longum viability in flan, but also increased their antiviral activity; potent anti-rotavirus effect of C. sorokiniana alone was observed. Although more studies are needed, results suggest that incorporation of this microalga into a dairy product confers enhanced viability and antiviral effects, which indicates that C. sorokiniana might be used as an ingredient to design products with additional health benefits.

Primary Dengue Infection in Patients Requiring Hospitalization During an Outbreak in a Low Incidence Mexican Region.

Background: Dengue manifestations can range from subclinical to fatal. The study of factors that influence dengue's clinical severity can provide information to potentially limit or predict severe cases. Secondary infection (SI) with a different dengue serotype has been recognized as an important determinant of severity. However, severe dengue (SD) manifestations, including shock, can happen during primary infection (PI) too and the mechanisms involved are less understood. To characterize the severe manifestations associated to PI, we distinguished between primary and secondary dengue cases in hospitalized patients from a region of low and recent dengue incidence in central Mexico. This region can serve as a model for dengue's behavior as it spreads to new areas worldwide. Methods: Dengue-specific immunoglobulin M (IgM) and IgG concentrations were measured in the serum of 78 hospitalized patients with dengue hemorrhagic fever, and their ratios were used to discriminate between PI and SI, as recommended by World Health Organization. Clinical and laboratory manifestations were compared between PI and SI. Results and Conclusions: PI was detected in 23% of hospitalized dengue cases, a proportion similar to that reported in high-incidence regions in Mexico. PI was more frequent in 16- to 40-year-olds, and was absent in patients older than 60 years. Only dengue with warning signs and SD were present in the studied population of hospitalized patients, and case frequency decreased with clinical severity both in PI and SI groups. No significant differences in demographics, laboratory tests, or symptoms were found between PI and SI, which illustrates that cases requiring hospitalization during outbreaks can be severe, even if they are PI. This information can help plan for sanitary contingencies in places where dengue is recently emergent and numerous PI cases are expected. The mechanisms involved in PI clinical severity need to be studied further.

Antiviral and immunomodulatory effects of polyphenols on macrophages infected with dengue virus serotypes 2 and 3 enhanced or not with antibodies.

Background: There is a lack of specific antiviral therapy against dengue virus (DENV) in current use. Therefore, a great proportion of dengue cases progress to severe clinical forms due to a complex interplay between virus and host immune response. It has been hypothesized that heterotypic non-neutralizing antibodies enhance DENV infection in phagocytic cells, and this induces an inflammatory response that is involved in the pathogenesis of severe dengue. Purpose: To identify the antiviral and immunomodulatory effects of polyphenols on dengue virus infection. Methods: Human U937-DC-SIGN macrophages were infected with DENV serotypes 2 or 3 in the presence or not of enhancing antibody 4G2. Viral titers and the secretion of tumor necrosis factor-alpha, IL-6, IL-10 and interferon-alpha were analyzed timely. Results: DENV infection alone induced high production of IL-6 and TNF-α, but in the presence of 4G2 antibody, viral titers and TNF-α secretion were potentiated. Based on anti-inflammatory antecedents, the polyphenols curcumin, fisetin, resveratrol, apigenin, quercetin and rutin were tested for antiviral and immunomodulatory properties. Only quercetin and fisetin inhibited DENV-2 and DENV-3 infection in the absence or presence of enhancing antibody (>90%, p<0.001); they also inhibited TNF-α and IL-6 secretion (p<0.001). Conclusion: Quercetin and fisetin down-regulate the production of proinflammatory cytokines induced by DENV infection enhanced by antibodies a mechanism involved in severe dengue.

Viral agents of gastroenteritis and their correlation with clinical symptoms in rotavirus-vaccinated children.

BACKGROUND AND OBJECTIVES: Enteric viral infections are among the leading causes of gastroenteritis in children up to five years of age worldwide. This study was aimed to determine the disease severity, incidence, and molecular genotyping of rotaviruses, noroviruses, astroviruses, and enteric adenoviruses as gastroenteritis agents among children up to five years old. MATERIALS AND METHODS: Gastroenteritis severity was determined by using the Ruuska and Vesikari score, whereas the incidence of enteric infections and their genotyping were determined by reverse transcription-polymerase chain reaction (RT-PCR) and sequence analysis. RESULTS: Rotaviruses were observed to possess the highest incidence with 10% (18/179) of the cases positives; nevertheless, noroviruses had the highest severe gastroenteritis score (13 ±â€¯3 points). Results indicated that 56% (10/18) of the detected rotavirus strains were genotype G12P[8], 50% (4/8) of noroviruses were GII.4 and 25% (2/8) were genotype GI.8. Out of the sapovirus positive samples, 30% (2/6) were genotyped as GI·I and GII·I. Sixty percent of the astrovirus strains (3/5) were genotype HAstV-2, and 20% (1/5) were genotype HAstV-6. Additionally, one of the adenovirus strains was identified as human mastadenovirus C type 6 specie. CONCLUSIONS: The diarrhea severity reduction in children provides evidence that the rotavirus vaccination program in the northwest of Mexico has been successful, even among children infected by the rotavirus emergent strain G12, however, norovirus resulted as the leading severe gastroenteritis-causing agent in children with rotavirus vaccine.

Assessment of the Sensitizing Potential of Proteins in BALB/c Mice: Comparison of Three Protocols of Intraperitoneal Sensitization.

Most food allergy cases are associated with a limited group of allergens. This could be attributed to an increased ability of some foods to sensitize and trigger allergic reactions. However, there are no validated animal models to evaluate the sensitizing or allergenic potentials of proteins. Our aim was to evaluate three protocols of adjuvant-free intraperitoneal sensitization that differ in the time points for sample collection (days 14, 28 and 35 from beginning of the sensitization) and also in the number of immunizations (2, 5 and 3, respectively). Ovalbumin (OVA; 0.05 mg), cow milk proteins (CMP; 0.025, 0.05 and 0.25 mg), and potato acid phosphatase (PAP; low allergenic protein; 250.0 mg) were administered intraperitoneally (ip) to BALB/c mice (n = 4⁻6) and the protein-specific IgE and IgG antibody responses were evaluated using ELISA. Additional serum protein-specific IgE antibodies evaluations were carried out after IgG depletion. Anti-OVA IgE antibodies were detected in mice from all three protocols. The responses were higher in the group of mice that underwent the 28-day protocol than in those that underwent the 14- or 35-day protocols (p < 0.01 and p < 0.05, respectively). Anti-CMP IgE antibodies were detected in both the 14- and 28-day protocols, but the response was higher in the group that underwent the 28-day protocol (p < 0.001). The anti-CMP IgE antibody response detection was improved after serum IgG depletion (p < 0.001). Anti-PAP IgE antibodies were not detected. Mice with undetectable serum levels of protein-specific IgE triggered anti-OVA, -CMP, and -PAP IgG responses. An adjuvant-free 28-day protocol with five ip immunizations seems appropriate for evaluation of the inherent sensitizing or allergenic capacity of the studied proteins. Reproducible results were obtained utilizing the BALB/c mouse strain. Inter-laboratory studies including a larger number of proteins should be carried out to validate this model.

La sucralosa promueve la polarización a macrófagos proinflamatorios M1

RESUMEN La sucralosa es un edulcorante no calórico de amplio consumo a nivel mundial, es considerado como un aditivo seguro, debido a que es eliminado en periodos cortos de tiempo. Recientemente se evidenció su bioacumulación en tejido adiposo, donde se encuentran inmersos macrófagos, células del sistema inmune involucradas en el desarrollo de la inflamación sistémica de bajo grado. A la fecha, no se cuenta con suficiente información para demostrar si los edulcorantes potencian los procesos inflamatorios alterando la función de células presentes en tejido y/o contribuyen en el desarrollo de patologías metabólicas. Por lo anterior, en nuestro trabajo se evaluó el efecto de la sucralosa en la viabilidad de los macrófagos diferenciados de la línea celular monocítica THP-1, por azul de tripán y ensayos de MTT, así como su efecto en la polarización M1/M2 por PCR según la expresión de IRF4, IRF5, STAT1, STAT6, perfil de expresión de IL-6, IL-12, TNF-α, TGF-β, IL-10 y SOCS3 por qPCR, y la cuantificación de la quimiocina IP-10 por ELISA. Los resultados indicaron que la sucralosa no tiene efectos citotóxicos, pero disminuye el número de células viables metabólicamente activas determinadas por MTT de manera dependiente de la concentración. La sucralosa incrementa la concentración de la quimiocina IP-10 y la expresión génica del factor de transcripción IRF5 y disminuye la expresión de IRF4 y STAT6, favoreciendo la polarización hacia poblaciones M1. La bioacumulación de sucralosa en tejido adiposo, y su interacción con macrófagos, podría inducir su polarización a M1.

ABSTRACT Sucralose is a non-nutritive sweetener widely consumed worldwide; it is considered a safe additive because it is eliminated quickly. Recently its bioaccumulation in adipose tissue was evidenced, where macrophages, cells of the immune system involved in developing low-grade systemic inflammation, are found. To date, there is a paucity of information regarding whether sweeteners potentiate inflammatory processes by altering the function of cells present in tissue and/or contribute to the development of metabolic pathologies. We evaluate the effect of sucralose on the viability of differentiated macrophages of the monocytic cell line THP-1, by trypan blue and MTT assays, respectively, as well as its effect on M1/ M2 by PCR according to the expression of IRF4, IRF5, STAT1, STAT6, expression profile of IL6, IL-12, TNF-α, TGF-β, IL-10 and SOCS3 by qPCR, and the quantification of the chemokine IP-10 by ELISE. The results indicated that sucralose has no cytotoxic effects but decreases the number of metabolically active viable cells determined by MTT of macrophages in a concentration-dependent manner. Sucralose increased the concentration of the chemokine IP-10 and the gene expression of the transcription factors IRF5 and decreased the expression of IRF4 and STAT 6 gene expression, favoring polarization towards M1 populations. The bioaccumulation of sucralose in adipose tissue, and its interaction with macrophages, could induce its polarization to M1.

Upregulation of the Suppressors of Cytokine Signaling 1 and 3 Is Associated with Arrest of Phosphorylated-STAT1 Nuclear Importation and Reduced Innate Response in Denguevirus-Infected Macrophages.

To clarify whether the suppressors of cytokine signaling (SOCS) are associated with denguevirus (DENV) evasion of the antiviral response, we analyzed the expression kinetics of SOCS1 and SOCS3 and of the antiviral genes MxA and OAS during DENV infection of U937 macrophages that were or not treated with interferon (IFN)-α. DENV infection produced a viral titer three times higher in untreated than in IFN-α-treated cells (p < 0.001 at 72 h postinfection [p.i.]). Partial inhibition of DENV replication was associated with reduced expression of MxA and OAS antiviral genes as well as higher SOCS1 and SOCS3 expression in DENV-infected cells than in cells treated only with IFN-α. Complete loss of phosphorylated-signal transducer and activator of transcription (p-STAT)2 and reduced nuclear importation of p-STAT1 were observed in DENV-infected cells compared to IFN-α treatment that induced p-STAT1 and p-STAT2. Our data thus suggest that overexpression of SOCS1 and SOCS3 induced by DENV infection leads to impairment of antiviral response through the inhibition of STAT functionality.

Female breast cancer incidence and mortality in Mexico, 2000-2010.

The objective of this study was to investigate the recent incidence and mortality trends for breast cancer in Mexican females. Data between 2000 and 2010 from the Department of Epidemiology of the Ministry of Health, and International Agency for Research on Cancer (IARC) were analyzed. Age-standardized rates (ASRs) and annual percent changes (APCs) were calculated. The absolute incidence and mortality rates of breast cancer increased: 3,726 and 4,615 in 2000 to 8,545 and 4,966 in 2010, respectively. Incidence increased over time in all age groups tested, the 60-64 age group had the highest ASR (57.4 per 100,000 women in 2010), while the 20-44 age group had the lowest ASR (12.3 in 2010). The results show that incidence of breast cancer has increased in Mexico during last one decade, especially among older women, while the downturn observed in mortality mainly reflects improved survival as a result of earlier diagnosis and better cancer treatment.