Gut Microbiota Composition Correlates with Disease Severity in Myelodysplastic Syndrome.

The myelodysplastic syndrome (MDS) is a heterogeneous group of clonal disorders of hematopoietic progenitor cells related to ineffective hematopoiesis and an increased risk of transformation to acute myelogenous leukemia. MDS is divided into categories, namely lineage dysplasia (MDS-SLD), MDS with ring sideroblasts (MDS-RS), MDS with multilineage dysplasia (MDS-MLD), MDS with excess blasts (MDS-EB). The International Prognostic Classification System (IPSS) ranks the patients as very low, low, intermediate, high, and very high based on disease evolution and survival rates. Evidence points to toll-like receptor (TLR) abnormal signaling as an underlying mechanism of this disease, providing a link between MDS and immune dysfunction. Microbial signals, such as lipopolysaccharides from gram-negative bacteria, can activate or suppress TLRs. Therefore, we hypothesized that MDS patients present gut microbiota alterations associated with disease subtypes and prognosis. To test this hypothesis, we sequenced the 16S rRNA gene from fecal samples of 30 MDS patients and 16 healthy elderly controls. We observed a negative correlation between Prevotella spp. and Akkermansia spp. in MDS patients compared with the control group. High-risk patients presented a significant increase in the genus Prevotella spp. compared to the other risk categories. There was a significant reduction in the abundance of the genus Akkermansia spp. in high-risk patients compared with low- and intermediate-risk. There was a significant decrease in the genus Ruminococcus spp. in MDS-EB patients compared with controls. Our findings show a new association between gut dysbiosis and higher-risk MDS, with a predominance of gram-negative bacteria.

Hypomethylation at H19DMR in penile squamous cell carcinoma is not related to HPV infection.

Penile squamous cell carcinoma (SCC) is a rare and aggressive tumour mainly related to lifestyle behaviour and human papillomavirus (HPV) infection. Environmentally induced loss of imprinting (LOI) at the H19 differentially methylated region (H19DMR) is associated with many cancers in the early events of tumorigenesis and may be involved in the pathogenesis of penile SCC. We sought to evaluate the DNA methylation pattern at H19DMR and its association with HPV infection in men with penile SCC by bisulfite sequencing (bis-seq). We observed an average methylation of 32.2% ± 11.6% at the H19DMR of penile SCC and did not observe an association between the p16INK4a+ (p = 0.59) and high-risk HPV+ (p = 0.338) markers with methylation level. The average methylation did not change according to HPV positive for p16INK4a+ or hrHPV+ (35.4% ± 10%) and negative for both markers (32.4% ± 10.1%) groups. As the region analysed has a binding site for the CTCF protein, the hypomethylation at the surrounding CpG sites might alter its insulator function. In addition, there was a positive correlation between intense polymorphonuclear cell infiltration and hypomethylation at H19DMR (p = 0.035). Here, we report that hypomethylation at H19DMR in penile SCC might contribute to tumour progression and aggressiveness regardless of HPV infection.

Synbiotic Sapota-do-Solimões (Quararibea cordata Vischer) Juice Improves Gut Microbiota and Short-Chain Fatty Acid Production in an In Vitro Model.

Sapota-do-Solimões (Quararibea cordata Vischer) is Amazon South América fruit found in Brazil, Colombia, Ecuador, and Peru. The orange-yellow fruit is usually eaten out of hand or as juice. Despite being a source of carotenoids and dietary fibers (pectin) that can reach the colon and act as an energy source for intestinal microbiota, the fruit is rarely known outside of South America. The symbiotic juice was prepared by fermenting the fruit juice with Lacticaseibacillus casei B-442 and adding prebiotic fructooligosaccharides (FOS, 7% w/v). This study evaluated the functional juice immediately after L. casei fermentation (SSJ0) and after 30 days of cold storage (SSJ30) regarding its effect on human colonic microbiota composition after in vitro fermentation. Fecal samples were collected from two healthy female volunteers, and the 16s rRNA gene sequencing analyzed the fecal microbiota composition. In vitro, colonic fermentation was performed using a batch bioreactor to simulate gastrointestinal conditions. The L. casei viability did not change significantly after 30 days of the synbiotic juice cold storage (4 °C). After the colonic fermentation, the relative abundance of Firmicutes decreased while Proteobacteria and Actinobacteria increased. Regarding short-chain fatty acid (SCFA) production by fecal colonic microbiota, the butyric acid was higher after sample SSJ0 fecal fermentation. In contrast, propionic, isobutyric, and acetic acids were higher after SSJ30 sample fecal fermentation. This study contributes to understanding the interactions between specific foods and the gut microbiota, which can affect human health and well-being.

Microbiota of healthy and bleached corals of the species Siderastrea stellata in response to river influx and seasonality in Brazilian northeast.

Although coral bleaching is increasing worldwide due to warming oceans exacerbated by climate change, there has been a growing recognition that local stressors may play an additional role. Important stressors include the physicochemical and microbiological influences that are related to river runoff. Here, we investigated the microbiota associated to mucus and tissue of endemic coral Siderastrea stellata, collected from Brazilian northeast coral reefs of Barra de Santo Antônio (subject to river runoff) and Maragogi (minimal river runoff) during both the rainy and dry seasons. We sequenced the V4 region of 16S rDNA and used multiple R packages to process raw data and performed statistical analysis to reveal the microbial community structure composition and functional predictions. Major dissimilarities between microbial communities were related to seasonality, while healthy and bleached specimens were mainly associated with the enrichment of several less abundant taxa involved in specific metabolic functions, mainly related to the nitrogen cycle. We were not able to observe the dominance of groups that has been previously associated with bleachings, such as Vibrionaceae or Burkholderiaceae. The influx of freshwater appears to increase the homogeneity between individuals in Barra de Santo Antonio, especially during the rainy season. By contrast, we observed an increased homogeneity between samples in Maragogi during the dry season. Understanding the dynamics of the coral microbiota and how bleaching appears in response to specific environmental variables, in addition to determining the conditions that lead to a more robust coral microbiota, is essential for choosing the most appropriate area and conservation methods, for example.

Impact of orange juice containing potentially prebiotic ingredients on human gut microbiota composition and its metabolites.

Orange juice containing oligosaccharides and dextran was assessed, after in vitro digestion, in a fermentative system with fecal microbiota from two donors (A and B) in anaerobiosis (37 °C/48 h). Microbiota analysis was performed using 16S rRNA sequencing, and HPLC quantified the metabolites. Oligosaccharides and dextran were resistant to digestion and were consumed during colonic fermentation. Lactate and acetate were the main metabolites, followed by butyrate, propionate, and isobutyrate. High propionate accumulation and fast consumption of oligosaccharides were observed for donor B. Lactobacillus ruminis (44.96 %) presented an increased relative abundance for donor A. Bifidobacterium adolescentis (41.73 %) was remarkable for donor B after fermentation. In addition, the functional orange juice promoted the growth of emerging probiotics such as Bacteroides xylanisolvens and reduced some strains, such as Clostridia sp. Thus, the potentially prebiotic orange juice enhances gut microbiota composition and might be a suitable product for the functional food market.

Multi-omic Analysis of the Gut Microbiome in Rats with Lithium-Pilocarpine-Induced Temporal Lobe Epilepsy.

Evidence supports that the gut microbiota and bacteria-dependent metabolites influence the maintenance of epileptic brain activity. However, the alterations in the gut microbiota between epileptic versus healthy individuals are poorly understood. We used a multi-omic approach to evaluate the changes in the composition of gut metagenome as well in the fecal metabolomic profile in rats before and after being submitted to status epilepticus (SE)-induced temporal lobe epilepsy (TLE). The 16S ribosomal RNA (rRNA) sequencing of fecal samples coupled to bioinformatic analysis revealed taxonomic, compositional, and functional shifts in epileptic rats. The species richness (Chao1 index) was significantly lower in the post-TLE group, and the ß-diversity analysis revealed clustering separated from the pre-TLE group. The taxonomic abundance analysis showed a significant increase of phylum Desulfobacterota and a decrease of Patescibacteria in the post-TLE group. The DESEq2 and LEfSe analysis resulted in 18 genera significantly enriched between post-TLE and pre-TLE groups at the genus level. We observed that epileptic rats present a peculiar metabolic phenotype, including a lower concentration of D-glucose and L-lactic acid and a higher concentration of L-glutamic acid and glycine. The microbiota-host metabolic correlation analysis showed that the genera differentially abundant in post-TLE rats are associated with the altered metabolites, especially the proinflammatory Desulfovibrio and Marvinbryantia, which were enriched in epileptic animals and positively correlated with these excitatory neurotransmitters and carbohydrate metabolites. Therefore, our data revealed a correlation between dysbacteriosis in epileptic animals and fecal metabolites that are known to be relevant for maintaining epileptic brain activity by enhancing chronic inflammation, an excitatory-inhibitory imbalance, and/or a metabolic disturbance. These data are promising and suggest that targeting the gut microbiota could provide a novel avenue for preventing and treating acquired epilepsy. However, the causal relationship between these microbial/metabolite components and the SRS occurrence still needs further exploration.

Contrasting modes of mitochondrial genome evolution in sister taxa of wood-eating marine bivalves (Teredinidae and Xylophagaidae).

The bivalve families Teredinidae and Xylophagaidae include voracious consumers of wood in shallow and deep-water marine environments, respectively. The taxa are sister clades whose members consume wood as food with the aid of intracellular cellulolytic endosymbionts housed in their gills. This combination of adaptations is found in no other group of animals and was likely present in the common ancestor of both families. Despite these commonalities, the two families have followed dramatically different evolutionary paths with respect to anatomy, life history and distribution. Here we present 42 new mitochondrial genome sequences from Teredinidae and Xylophagaidae and show that distinct trajectories have also occurred in the evolution and organization of their mitochondrial genomes. Teredinidae display significantly greater rates of amino acid substitution but absolute conservation of protein-coding gene order, whereas Xylophagaidae display significantly less amino acid change but have undergone numerous and diverse changes in genome organization since their divergence from a common ancestor. As with many bivalves, these mitochondrial genomes encode two ribosomal RNAs, 12 protein coding genes, and 22 tRNAs; atp8 was not detected. We further show that their phylogeny, as inferred from amino acid sequences of 12 concatenated mitochondrial protein-coding genes, is largely congruent with those inferred from their nuclear genomes based on 18S and 28S ribosomal RNA sequences. Our results provide a robust phylogenetic framework to explore the tempo and mode of mitochondrial genome evolution and offer directions for future phylogenetic and taxonomic studies of wood-boring bivalves.

True or false: what are the factors that influence COVID-19 diagnosis by RT-qPCR?

INTRODUCTION: The Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) disease has had a catastrophic impact on the world resulting in several deaths. Since World Health Organization declared the pandemic status of the disease, several molecular diagnostic kits have been developed to help the tracking of viruses spread. AREAS COVERED: This review aims to describe and evaluate the currently reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) diagnosis kit. Several processes used in COVID-19 diagnostic procedures are detailed in further depth to demonstrate optimal practices. Therefore, we debate the main factors that influence the viral detection of SARS-COV-2 and how they can affect the diagnosis of patients. EXPERT OPINION: Here is highlighted and discussed several factors that can interfere in the RT-PCR analysis, such as the viral load of the sample, collection site, collection methodology, sample storage, transport, primer, and probe mismatch/dimerization in different brand kits. This is a pioneer study to discuss the factor that could lead to the wrong interpretation of RT-qPCR diagnosis of SARS-CoV-2. This study aimed to help the readers to understand what very likely is behind a bad result of SARS-CoV-2 detection by RT-PCR and what could be done to reach a reliable diagnosis.

Effects of the antibiotics trimethoprim (TMP) and sulfamethoxazole (SMX) on granulation, microbiology, and performance of aerobic granular sludge systems.

This work assessed the effect of the antibiotics trimethoprim (TMP) and sulfamethoxazole (SMX) on the granulation process, microbiology, and organic matter and nutrient removal of an aerobic granular sludge (AGS) system. In addition, after the maturation stage, the impact of the redox mediator anthraquinone-2,6-disulfonate (AQDS) (25 µM) on the biotransformation of the antibiotics was evaluated. The reactor R1 was maintained as a control, and the reactor R2 was supplemented with TMP and SMX (200 µg L-1). The ability to remove C, N, and P was similar between the reactors. However, the structural integrity of the AGS was impaired by the antibiotics. Low TMP (∼30%) and SMX (∼60%) removals were achieved when compared to anaerobic or floccular biomass aerobic systems. However, when the system was supplemented with AQDS, an increase in the removal of TMP (∼75%) and SMX (∼95%) was observed, possibly due to the catalytic action of the redox mediator on cometabolic processes. Regarding the microbial groups, whereas Proteobacteria and Bacterioidetes increased, Planctomycetes decreased in both reactors. However, TMP and SMX presence seemed to inhibit or favor some genera during the formation of the granules, possibly due to their bactericidal action.