In vitro anti-tumoral and immunomodulatory effects of acidic polysaccharides isolated from the fungus Gloeosoma mirabile.

Polysaccharides from wood-rooting fungi have attracted attention due to their broad pharmacological properties. Herein, we report the antitumor and immunomodulatory activities of acid polysaccharides isolated from fungi Gloeosoma mirabile. The polysaccharide extracts displayed significant antiproliferative activity against cancer cell lines (MCF-7, HCT-116, U-937) in a dose-dependent manner and induction of IL-6 in macrophage RAW 264.7. Furthermore, flow cytometry analysis showed that high polysaccharide concentrations induced apoptosis by 83% in HL-60 cells. Based on gas chromatography-mass spectrometry (GC-MS) and Fourier transform infra-red (FT-IR) spectroscopy studies, acidic polysaccharides from G. mirabile were mainly composed of arabinose, α-D-galactopyranose and methyl ß-D-galactopyranoside.

A Systematic Review of Gene Expression Studies in Critically Ill Patients with Sepsis and Community-Acquired Pneumonia.

(1) Background: Sepsis is present in nearly 90% of critically ill patients with community-acquired pneumonia (CAP). This systematic review updates the information on studies that have assessed gene expression profiles in critically ill septic patients with CAP. (2) Methods: We searched for studies that satisfied the following criteria: (a) expression profile in critically ill patients with sepsis due to CAP, (b) presence of a control group, and (c) adult patients. Over-representation analysis was performed with clusterProfiler using the Hallmark and Reactome collections. (3) Results: A total of 4312 differentially expressed genes (DEGs) and sRNAs were included in the enrichment analysis. In the Hallmark collection, genes regulated by nuclear factor kappa B in response to tumor necrosis factor, genes upregulated by signal transducer and activator of transcription 5 in response to interleukin 2 stimulation, genes upregulated in response to interferon-gamma, genes defining the inflammatory response, a subgroup of genes regulated by MYC-version 1 (v1), and genes upregulated during transplant rejection were significantly enriched in critically ill septic patients with CAP. Moreover, 88 pathways were identified in the Reactome database. (4) Conclusions: This study summarizes the reported DEGs in critically ill septic patients with CAP and investigates their functional implications. The results highlight the complexity of immune responses during CAP.

In vitro cytotoxic capacity against tumor cell lines and antioxidant activity of acidic polysaccharides isolated from the Andean Patagonian fungus Phylloporia boldo.

Fungal polysaccharides possess a broad biological activity, including cytotoxic and antioxidant activities. This work aimed to evaluate the cytotoxic and antioxidant activity of the acidic polysaccharides of Phylloporia boldo strain (named PBAP40). Cytotoxic activity of polysaccharide was evaluated determining the viability of three tumor cell lines by MTT assay. The effect of acidic polysaccharide on the cell cycle of HL-60 cell line was evaluated by flow cytometry, and the antioxidant activity was determined by DPPH and ABTS assays. PBAP40 showed cytotoxic effects in tumor cell lines. Results suggest that P. boldo acidic polysaccharides arrested tumor cells in the cell cycle Sub G1 phase. The acidic polysaccharides of PBAP40 strain were not cytotoxic for the non-tumor cell line. PBAP40 also showed excellent antioxidant activity. The FT-IR analysis of the acidic polysaccharides indicated the presence of glucans bearing α- and ß- type glycosidic bonds.

Sulfated Polysaccharide Extracted from the Green Algae Codium bernabei: Physicochemical Characterization and Antioxidant, Anticoagulant and Antitumor Activity.

Codium bernabei is a green alga that grows on Chilean coasts. The composition of its structural polysaccharides is still unknown. Hence, the aim of this work is to isolate and characterize the hot water extracted polysaccharide fractions. For this purpose, the water extracts were further precipitated in alcohol (TPs) and acid media (APs), respectively. Both fractions were characterized using different physicochemical techniques such as GC-MS, GPC, FTIR, TGA, and SEM. It is confirmed that the extracted fractions are mainly made of sulfated galactan unit, with a degree of sulfation of 19.3% (TPs) and 17.4% (ATs) and a protein content of 3.5% in APs and 15.6% in TPs. Other neutral sugars such as xylose, glucose, galactose, fucose, mannose, and arabinose were found in a molar ratio (0.05:0.6:1.0:0.02:0.14:0.11) for TPs and (0.05:0.31:1.0:0.03:0.1:0.13) for ATs. The molecular weight of the polysaccharide samples was lower than 20 kDa. Both polysaccharides were thermally stable (Tonset > 190 °C) and showed antioxidant activity according to the ABTS•+ and DPPH tests, where TPs fractions had higher scavenging activity (35%) compared to the APs fractions. The PT and APTTS assays were used to measure the anticoagulant activity of the polysaccharide fractions. In general, the PT activity of the TPs and APs was not different from normal plasma values. The exception was the TPs treatment at 1000 µg mL−1 concentration. The APTTS test revealed that clotting time for both polysaccharides was prolonged regarding normal values at 1000 µg mL−1. Finally, the antitumor test in colorectal carcinoma (HTC-116) cell line, breast cancer (MCF-7) and human leukemia (HL-60) cell lines showed the cytotoxic effect of TPs and APs. Those results suggest the potential biotechnological application of sulfate galactan polysaccharides isolated from a Chilean marine resource.

N6 -Methyladenosine Negatively Regulates Human Respiratory Syncytial Virus Replication.

N6-methyladenosine (m6A) is the most abundant internal modification described in eukaryotic mRNA and several viral RNA including human respiratory syncytial virus (HRSV). Here, we evaluated the impact of m6A writers, erasers and readers on HRSV genomic RNA accumulation and inclusion bodies assembly during viral replication. We observed that the METTL3/METTL14 m6A writer complex plays a negative role in HRSV protein synthesis and viral titers, while m6A erasers FTO and ALKBH5 had the opposite effect. We also observed that m6A readers YTHDF1-3 bind to the viral genomic RNA inducing a decrease in its intracellular levels and thus, inhibiting viral replication. Finally, we observed that overexpression of YTHDFs proteins caused a decrease in the size of inclusion bodies (IBs), accompanied by an increase in their number. METTL3 knockdown cells showed an opposite effect indicating that the dynamics of IBs assembly and coalescence are strongly affected by m6A readers in a mechanism dependent on m6A writers. Taken together, our results demonstrated that the m6A modification negatively affects HRSV replication, possibly through a mechanism involving the assembly of inclusion bodies, the main factories of viral genomic RNA synthesis.

Diarrheagenic Escherichia coli pathotypes isolated from a swine farm in a region of Morelos state, Mexico / Patotipos diarreicos de Escherichia coli aislados de una granja porcina en una región del estado de Morelos, México

Abstract: Objective: Determine the frequency of diarrheagenic Escherichia coli pathotypes colonizing swine. Materials and methods: E. coli strains isolated of fecal samples from 280 swine, produced for local consumption, in a semi-technical farm of Morelos state (central Mexico), were tested to identify the diarrheagenic E. coli pathotypes by multiplex PCR. Pulsed-field gel electrophoresis (PFGE) were determined for the study of genetic diversity between pathigenic E. coli strains. Results: Of the 521-diarrheagenic E. coli isolates examined, 50 (9.6%) were positive for at least one virulence gene in 42 different animals. Thus, 15% (42/280) of the swine in this farm were colonized with pathogenic E. coli. Among the E. coli isolates, the pathotype EPEC (6.5%) was the most frequently, followed by EHEC (2.3%), ETEC and EIEC (0.4%). The study of genetic diversity, carried out by PFGE of 40 representative isolates, revealed 25 distinct restriction profiles clustered in 21 groups (A-U). Conclusions: In this study, four different E. coli pathotypes were found among swine colonized by E. coli in this farm. Thus, these swine are reservoirs for these virulent bacteria and there is potential risk of causing diarrhea in swine and in the population consuming the meat.

Resumen: Objetivo: Determinar la frecuencia de patotipos diarreicos de Escherichia coli que colonizan cerdos. Material y métodos: Se analizaron cepas de E. coli aisladas de muestras fecales de 280 cerdos, producidas para consumo local, en una granja del estado de Morelos (centro de México) para identificar los patotipos diarreicos de E. coli por PCR multiplex. Se determinó la electroforesis en gel de campo pulsado (PFGE) para el estudio de la diversidad genética entre cepas patógenas de E. coli. Resultados: De los 521 aislados de E. coli con patotipos diarreicos examinados, 50 (9.6%) fueron positivos para al menos un gen de virulencia en 42 animales diferentes. Así, 15% (42/280 porcinos) fueron colonizados con E. coli patógeno. Entre los aislados de E. coli de porcinos, el patotipo EPEC (6.5%) fue el más frecuente, seguido por EHEC (2.3%), ETEC y EIEC (0.4%). El estudio de la diversidad genética, realizado por PFGE de 40 cepas representativas, reveló 25 perfiles de restricción distintos agrupados en 21 grupos (A-U). Conclusiones: En este estudio, se encontraron cuatro diferentes patotipos de E. coli entre los cerdos colonizados por E. coli. Estos cerdos son reservorios de estas bacterias virulentas y existe un riesgo potencial de causar diarrea en los cerdos y en la población que consume la carne.

Características nutricionales y efecto citotóxico de polisacáridos extraídos de los digüeñes Cyttaria berteroi y Cyttaria hariotii presentes en Chile / Nutritional characteristics and cytotoxic effect of polysaccharides extracted from the digüeñes Cyttaria berteroi and Cyttaria hariotii present in Chile

RESUMEN El género Cyttaria pertenece a la familia Cyttariaceae, existen 11 especies descritas a nivel mundial. En Chile y Argentina podemos encontrar sólo siete de ellas, conocidas como "Digüeñes" del Mapudungun "diweñ", hongos parásitos obligados de árboles de Nothofagus. Este género, se distribuye naturalmente en el hemisferio sur y tiene gran importancia desde el punto de vista alimenticio. A pesar del extenso conocimiento sobre la taxonomía, ecología y composición químico-nutricional de algunas especies de Cyttaria, su potencial nutracéutico ha recobrado interés recientemente. En esta investigación, se determinó la composición químicaproximal y actividad citotóxica de los polisacáridos de C. berteroi y C. hariotii. Para caracterizar la composición nutricional de las especies de Cyttaria, se utilizó la metodología de AOAC y se avaluó el potencial citotóxico de sus polisacáridos mediante el ensayo MTT (bromuro de 3- (4,5-dimetiltiazol-2-il) -2,5-difeniltetrazolio) frente a las líneas celulares de cáncer de colon humano (HCT-116), leucemia humana (U-937) y cáncer de mama (MCF-7). Los resultados evidenciaron que C. berteroi presentó un alto contenido de proteínas y lípidos en comparación con C. hariotii. Además, los polisacáridos de C. hariotii tienen un mayor efecto citotóxico frente a la línea celular de leucemia (IC50= 2100 µg/mL), cáncer de colon (IC50= 3700 µg/mL) y cáncer de mama (IC50=9470 µg/mL). En consecuencia, se concluye que los metabolitos de C. berteroi y la actividad citotóxica en líneas tumorales de los polisacáridos de C. hariotii podrían representar una oportunidad para la obtención de un potencial producto nutracéutico.

ABSTRACT The Cyttaria genus belongs to the Cyttariaceae family, with 11 species described worldwide. In Chile and Argentina, seven of them are found. They are known as "Digüeñes", from the Mapudungun "diweñ", and are parasitic fungi of Nothofagus trees. They are naturally distributed in the southern hemisphere and are of great importance from a nutritional point of view. Despite extensive knowledge about the taxonomy, ecology, and chemical-nutritional composition of some Cyttaria species, their nutraceutical potential has recently gained interest. In this investigation, the chemical-proximal composition and cytotoxic activity of the C. berteroi and C. hariotii polysaccharides were determined. To identify the nutritional composition of the Cyttaria species, the AOAC methodology was used and the cytotoxic potential of their polysaccharides was evaluated by means of the MTT test (3- (4,5-dimethylthiazol-2-yl) -2,5 bromide-diphenyltetrazolium) against the cell lines of human colon cancer (HCT-116), human leukemia (U-937) and breast cancer (MCF-7). The results showed that C. berteroi had a high protein and lipid content compared to C. hariotii. Also, C. hariotii polysaccharides had a greater cytotoxic effect against the colon cancer cell line (IC50= 3700 µg/mL), leukemia (IC50= 2100 µg/mL) and breast cancer (IC50= 9470 µg/mL). Therefore, it is concluded that C. berteroi metabolites and cytotoxic activity in C. hariotii polysaccharide tumor lines could represent an opportunity to obtain a potential nutraceutical product.

Diarrheagenic Escherichia coli pathotypes isolated from a swine farm in a region of Morelos state, Mexico.

OBJECTIVE: Determine the frequency of diarrheagenic Escherichia coli pathotypes colonizing swine. MATERIALS AND METHODS: E. coli strains isolated of fecal samples from 280 swine, produced for local consumption, in a semi-technical farm of Morelos state, (central Mexico) were tested to identify the diarrheagenic E. coli pathotypes by multiplex PCR. RESULTS: Of the 521-diarrheagenic E. coli isolates examined, 50 (9.6%) were positive for at least one virulence gene in 42 different animals. Thus, 15% (42/280) of the swine in this farm were colonized with pathogenic E. coli. Among the E. coli isolates, the pathotype EPEC (6.5%) was the most frequently, followed by EHEC (2.3%), ETEC and EIEC (0.4%). CONCLUSIONS: In this study, four different E. coli pathotypes were found among swine colonized by E. coli in this farm. Thus, these swine are reservoirs for these virulent bacteria and there is potential risk of causing diarrhea in swine and in the population consuming the meat.