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.

Efecto aditivo in vitro de Chlorella sorokiniana en combinación con Vincristina sobre la inhibición del crecimiento de células de cáncer de colon HT-29 / In vitro additive effect of Chlorella sorokiniana in combination withVincristine on HT-29 colon cancer cells growth inhibition

Objetivo: Evaluar el efecto in vitro de la combinación deChlorella sorokiniana con Vincristina contra el crecimiento decélulas de cáncer de colon HT-29.Material y método:Chlorella sorokiniana se cultivó enmedio López-Chuken. El efecto inhibitorio de la microalga solay en combinación con Vincristina en el crecimiento tumoral seevaluó mediante la técnica de MTT, contra células de cáncerde colon humano HT-29, y se analizó mediante el softwareSynergyFinder 2.0.Resultados: El crecimiento Chlorella sorokiniana fue cons-tante al día 28 a una temperatura de 34 oC ± 3 oC. El efectoinhibitorio de Vincristina sobre células HT-29 fue del 60% apartir de 0.0037μg/mL. La inhibición por Chlorella sorokinianafue del 60% al 80% a las concentraciones de 106-108.Además, la combinación de Vincristina/Chlorella inhibió el cre-cimiento tumoral entre 70% y 90%, siendo la concentraciónmenor de Chlorella la que mostró un mejor efecto en combi-nación con Vincristina. El análisis de los resultados enSynergyFinder mostró un score de -0.708, determinando unefecto aditivo. Conclusión:Chlorella sorokiniana presenta un efecto adi-tivo en combinación con Vincristina contra la línea de cáncerde colon humano HT-29. La suplementación de C. sorokinianaen la dieta de pacientes con cáncer de colon podría mejorarsu tratamiento y por consecuencia su recuperación (AU)

Objective:To evaluate in vitro the effect of the combina-tion of Chlorella sorokiniana with vincristine on HT-29 coloncancer cells.Material and method:Chlorella sorokininana growth wasconstant on day 28 at a temperature of 34 oC ± 3 oC. Chlorellasorokiniana was cultured in López-Chuken medium. HT-29 cellsgrowth inhibition by the microalga alone or in combination withvincristine was evaluated by the colorimetric reduction MTT as-say, and analyzed using the SynergyFinder 2.0 software. Results:The inhibitory effect of Vincristine on HT-29 cellswas 60% from 0.0037μg/mL. Tumor cells growth inhibition by106 to 108 Chlorella sorokiniana cells ranged from 60% to80%. The combination of vincristine and Chlorella inhibitedtumor cells growth from 70% to 90%, being the lower con-centration of Chlorella the one that showed a better effect incombination with vincristine. The analysis of the results inSynergyFinder showed a score of -0.708, determining an ad-ditive effect.Conclusion: Chlorella sorokiniana has an additive effect incombination with vincristine against the human colon cancerline HT-29. Supplementation of C. sorokiniana in the diet ofpatients with colon cancer may improve their treatment andrecovery (AU)

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.

Molecular detection of equine infectious anemia virus in clinically normal, seronegative horses in an endemic area of Mexico.

Equine infectious anemia (EIA) is a highly infectious disease in members of the Equidae family, caused by equine infectious anemia virus (EIAV). The disease severity ranges from subclinical to acute or chronic, and causes significant economic losses in the equine industry worldwide. Serologic tests for detection of EIAV infection have some concerns given the prolonged seroconversion time. Therefore, molecular methods are needed to improve surveillance programs for this disease. We attempted detection of EIAV in 6 clinical and 42 non-clinical horses in Nuevo Leon State, Mexico, using the agar gel immunodiffusion (AGID) test for antibody detection, and nested and hemi-nested PCR for detection of proviral DNA. We found that 6 of 6, 5 of 6, and 6 of 6 clinical horses were positive by AGID, nested PCR, and hemi-nested PCR, respectively, whereas 0 of 42, 1 of 42, and 9 of 42 non-clinical horses were positive by these tests, respectively. BLAST analysis of the 203-bp 5'-LTR/tat segment of PCR product revealed 83-93% identity with EIAV isolates in GenBank and reference strains from other countries. By phylogenetic analysis, our Mexican samples were grouped in a different clade than other sequences reported worldwide, indicating that the LRT/tat region represents an important target for the detection of non-clinical horses.

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.

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.

Bioactive Products From Plant-Endophytic Gram-Positive Bacteria.

Endophytes constitute plant-colonizing microorganisms in a mutualistic symbiosis relationship. They are found in most ecosystems reducing plant crops' biotic and abiotic stressors by stimulating immune responses, excluding plant pathogens by niche competition, and participating in antioxidant activities and phenylpropanoid metabolism, whose activation produces plant defense, structural support, and survival molecules. In fact, metabolomic studies have demonstrated that endophyte genes associated to specific metabolites are involved in plant growth promotion (PGP) by stimulating plant hormones production such as auxins and gibberellins or as plant protective agents against microbial pathogens, cancer, and insect pests, but eco-friendly and eco-safe. A number of metabolites of Gram-positive endophytes isolated from agriculture, forest, mangrove, and medicinal plants, mainly related to the Firmicutes phyla, possess distinctive biocontrol and plant growth-promoting activities. In general, Actinobacteria and Bacillus endophytes produce aromatic compounds, lipopeptides, plant hormones, polysaccharides, and several enzymes linked to phenylpropanoid metabolism, thus representing high potential for PGP and crop management strategies. Furthermore, Actinobacteria have been shown to produce metabolites with antimicrobial and antitumor activities, useful in agriculture, medicine, and veterinary areas. The great endophytes diversity, their metabolites production, and their adaptation to stress conditions make them a suitable and unlimited source of novel metabolites, whose application could reduce agrochemicals usage in food and drugs production.

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.

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.

Betaine Aldehyde Dehydrogenase expression during physiological cardiac hypertrophy induced by pregnancy.

Betaine Aldehyde Dehydrogenase (betaine aldehyde: NAD(P)+ oxidoreductase, (E.C. 1.2.1.8; BADH) catalyze the irreversible oxidation of betaine aldehyde (BA) to glycine betaine (GB) and is essential for polyamine catabolism, γ-aminobutyric acid synthesis, and carnitine biosynthesis. GB is an important osmolyte that regulates the homocysteine levels, contributing to a vascular risk factor reduction. In this sense, distinct investigations describe the physiological roles of GB, but there is a lack of information about the GB novo synthesis process and regulation during cardiac hypertrophy induced by pregnancy. In this work, the BADH mRNA expression, protein level, and activity were quantified in the left ventricle before, during, and after pregnancy. The mRNA expression, protein content and enzyme activity along with GB content of BADH increased 2.41, 1.95 and 1.65-fold respectively during late pregnancy compared to not pregnancy, and returned to basal levels at postpartum. Besides, the GB levels increased 1.53-fold during pregnancy and remain at postpartum. Our results demonstrate that physiological cardiac hypertrophy induced BADH mRNA expression and activity along with GB production, suggesting that BADH participates in the adaptation process of physiological cardiac hypertrophy during pregnancy, according to the described GB role in cellular osmoregulation, osmoprotection and reduction of vascular risk.

HIF-1α and PPARγ during physiological cardiac hypertrophy induced by pregnancy: Transcriptional activities and effects on target genes.

Hypoxia inducible factor 1-α (HIF-1α) and peroxisome proliferator-activated receptor γ (PPARγ) are transcription factors that activate genes involved in cellular metabolism. Physiological cardiac hypertrophy induced by pregnancy initiates compensatory changes in metabolism. However, the contributions of HIF-1α and PPARγ to this physiological status and to its reversible, metabolic process (postpartum) in the heart are not well-defined. Therefore, the aim of the present study was to evaluate the transcriptional activities of HIF-1α and PPARγ in the left ventricle of rats before, during, and after pregnancy. Furthermore, the effects of pregnancy on target genes of glycolysis and glycerol-lipid biosynthesis, key regulatory enzymes, and metabolic intermediates were evaluated. The activities of HIF-1α and PPARγ increased 1.2- and 1.6-fold, respectively, during pregnancy, and decreased to basal levels during postpartum. Expressions of mRNA for glucose transport 1 (GLUT1), enzymes of glycolysis (HK2, PFKM, and GAPDH) and glycerol-lipid biosynthesis (GPAT and GPD1) increased 1.6- to 14-fold during pregnancy and returned to basal levels postpartum. The increase in GPD1 expression translated to an increase in its activity, but such was not the case for GAPDH suggesting that post-translational regulation of these proteins is differential during pregnancy. Glycolytic (glucose, lactate, and DHAP) and glycerol-lipid biosynthesis (G3P and FFA) intermediates increased with pregnancy and were maintained postpartum. The results demonstrate that pregnancy-induced, physiological cardiac hypertrophy activates the expression of genes involved in glycolytic and glycerol-lipid biosynthesis suggesting that the shift in cardiac metabolism is mediated by the activation of HIF-1α and PPARγ.

Mutation distribution in the NSP4 protein in rotaviruses isolated from Mexican children with moderate to severe gastroenteritis.

The NSP4 protein is a multifunctional protein that plays a role in the morphogenesis and pathogenesis of the rotavirus. Although NSP4 is considered an enterotoxin, the relationship between gastroenteritis severity and amino acid variations in NSP4 of the human rotavirus remains unclear. In this study, we analyzed the sequence diversity of NSP4 and the severity of gastroenteritis of children with moderate to severe gastroenteritis. The rotavirus-infected children were hospitalized before the rotavirus vaccine program in Mexico. All children had diarrhea within 1-4 days, 44 (88%) were vomiting and 35 (70%) had fevers. The severity analysis showed that 13 (26%) cases had mild gastroenteritis, 23 (46%) moderate gastroenteritis and 14 (28%) severe. NSP4 phylogenetic analysis showed three clusters within the genotype E1. Sequence analysis revealed similar mutations inside each cluster, and an uncommon variation in residue 144 was found in five of the Mexican NSP4 sequences. Most of the amino acid variations were located in the VP4 and VP6 binding site domains, with no relationship to different grades of gastroenteritis. This finding indicates that severe gastroenteritis caused by the rotavirus appears to be related to diverse viral or cellular factors instead of NSP4 activity as a unique pathogenic factor.