Fecal microbiota transplantation: current challenges and future landscapes.

SUMMARYGiven the importance of gut microbial homeostasis in maintaining health, there has been considerable interest in developing innovative therapeutic strategies for restoring gut microbiota. One such approach, fecal microbiota transplantation (FMT), is the main "whole gut microbiome replacement" strategy and has been integrated into clinical practice guidelines for treating recurrent Clostridioides difficile infection (rCDI). Furthermore, the potential application of FMT in other indications such as inflammatory bowel disease (IBD), metabolic syndrome, and solid tumor malignancies is an area of intense interest and active research. However, the complex and variable nature of FMT makes it challenging to address its precise functionality and to assess clinical efficacy and safety in different disease contexts. In this review, we outline clinical applications, efficacy, durability, and safety of FMT and provide a comprehensive assessment of its procedural and administration aspects. The clinical applications of FMT in children and cancer immunotherapy are also described. We focus on data from human studies in IBD in contrast with rCDI to delineate the putative mechanisms of this treatment in IBD as a model, including colonization resistance and functional restoration through bacterial engraftment, modulating effects of virome/phageome, gut metabolome and host interactions, and immunoregulatory actions of FMT. Furthermore, we comprehensively review omics technologies, metagenomic approaches, and bioinformatics pipelines to characterize complex microbial communities and discuss their limitations. FMT regulatory challenges, ethical considerations, and pharmacomicrobiomics are also highlighted to shed light on future development of tailored microbiome-based therapeutics.

Fecal-adherent mucus is a non-invasive source of primary human MUC2 for structural and functional characterization in health and disease.

The O-glycoprotein Mucin-2 (MUC2) forms the protective colon mucus layer. While animal models have demonstrated the importance of Muc2, few studies have explored human MUC2 in similar depth. Recent studies have revealed that secreted MUC2 is bound to human feces. We hypothesized human fecal MUC2 (HF-MUC2) was accessible for purification and downstream structural and functional characterization. We tested this via histologic and quantitative imaging on human fecal sections; extraction from feces for proteomic and O-glycomic characterization; and functional studies via growth and metabolic assays in vitro. Quantitative imaging of solid fecal sections showed a continuous mucus layer of varying thickness along human fecal sections with barrier functions intact. Lectin profiling showed HF-MUC2 bound several lectins but was weak to absent for Ulex europaeus 1 (α1,2 fucose-binding) and Sambucus nigra agglutinin (α2,6 sialic acid-binding), and did not have obvious b1/b2 barrier layers. HF-MUC2 separated by electrophoresis showed high molecular weight glycoprotein bands (∼1-2 MDa). Proteomics and Western analysis confirmed the enrichment of MUC2 and potential MUC2-associated proteins in HF-MUC2 extracts. MUC2 O-glycomics revealed diverse fucosylation, moderate sialylation, and little sulfation versus porcine colonic MUC2 and murine fecal Muc2. O-glycans were functional and supported the growth of Bacteroides thetaiotaomicron (B. theta) and short-chain fatty acid (SCFA) production in vitro. MUC2 could be similarly analyzed from inflammatory bowel disease stools, which displayed an altered glycomic profile and differential growth and SCFA production by B. theta versus healthy samples. These studies describe a new non-invasive platform for human MUC2 characterization in health and disease.

Genetic architecture and correlations between the gut microbiome and gut gene transcription in Chinook salmon (Oncorhynchus tshawytscha).

Population divergence through selection can drive local adaptation in natural populations which has implications for the effective restoration of declining and extirpated populations. However, adaptation to local environmental conditions is complicated when both the host and its associated microbiomes must respond via co-evolutionary change. Nevertheless, for adaptation to occur through selection, variation in both host and microbiome traits should include additive genetic effects. Here we focus on host immune function and quantify factors affecting variation in gut immune gene transcription and gut bacterial community composition in early life-stage Chinook salmon (Oncorhynchus tshawytscha). Specifically, we utilized a replicated factorial breeding design to determine the genetic architecture (sire, dam and sire-by-dam interaction) of gut immune gene transcription and microbiome composition. Furthermore, we explored correlations between host gut gene transcription and microbiota composition. Gene transcription was quantified using nanofluidic qPCR arrays (22 target genes) and microbiota composition using 16 S rRNA gene (V5-V6) amplicon sequencing. We discovered limited but significant genetic architecture in gut microbiota composition and transcriptional profiles. We also identified significant correlations between gut gene transcription and microbiota composition, highlighting potential mechanisms for functional interactions between the two. Overall, this study provides support for the co-evolution of host immune function and their gut microbiota in Chinook salmon, a species recognized as locally adapted. Thus, the inclusion of immune gene transcription profile and gut microbiome composition as factors in the development of conservation and commercial rearing practices may provide new and more effective approaches to captive rearing.

Effect of Astigmatism Treatment With Three Excimer Lasers After Photorefractive Keratectomy.

OBJECTIVES: To compare astigmatism correction after photorefractive keratectomy (PRK) using three excimer laser devices, Alcon Wavelight EX500, Schwind Amaris 1,050, and Technolas Teneo317 M2. METHODS: This retrospective study included 414 eyes from 414 subjects with a history of PRK performed using Alcon Wavelight (n=172), Schwind Amaris (n=122), and Technolas (n=120). Uncorrected & corrected distance visual acuity (UDVA & CDVA), and refractive status (spherical equivalent (SE) and vector analysis (J0 and J45)) were postoperatively recorded at the 6-month and 12-month examinations. RESULTS: The mean CDVA and J45 at 6-month and 12-month showed no statistically significant difference among the three different excimer lasers ( P >0.05). There was a statistically significant difference in mean UDVA and J0 at 6-month and 12-month after PRK among the three groups, with no statistically significant difference between Alcon Wavelight and Schwind Amaris lasers, while both of the prior lasers showed a significant difference with the Technolas laser. The highest and lowest changes in the magnitude of J0 in 6-month and 12-month follow-ups were seen for the Alcon Wavelight and Technolas groups, respectively. Both Schwind Amaris and Technolas had a small hyperopic SE while the Alcon Wavelight's SE was minimally myopic. This difference in SE between Alcon Wavelight and both Schwind Amaris and Technolas reached statistical significance. CONCLUSION: While all three lasers performed well in reducing preoperative astigmatism; however, the Alcon Wavelight and Schwind Amaris were more effective in correcting astigmatism than the Technolas. The difference between the Alcon Wavelight and Schwind Amaris did not reach statistical significance.

Behavioral transcriptomic effects of triploidy and probiotic therapy (Bifidobacterium, Lactobacillus, and Lactococcus mixture) on juvenile Chinook salmon (Oncorhynchus tshawytscha).

Aquaculturists use polyploid fish to maximize production albeit with some unintended consequences including compromised behaviors and physiological function. Given benefits of probiotic therapies (e.g., improved immune response, growth, and metabolism), we explored probiotic supplementation (mixture of Bifidobacterium, Lactobacillus, and Lactococcus), to overcome drawbacks. We first examined fish gut bacterial community composition using 16S metabarcoding (via principal coordinate analyses and PERMANOVA) and determined probiotics significantly impacted gut bacteria composition (p = 0.001). Secondly, we examined how a genomic disruptor (triploidy) and diet supplements (probiotics) impact gene transcription and behavioral profiles of hatchery-reared Chinook salmon (Oncorhynchus tshawytscha). Juveniles from four treatment groups (diploid-regular feed, diploid-probiotic feed, triploid-regular feed, and triploid-probiotic feed; n = 360) underwent behavioral assays to test activity, exploration, neophobia, predator evasion, aggression/sociality, behavioral sensitivity, and flexibility. In these fish, transcriptional profiles for genes associated with neural functions (neurogenesis/synaptic plasticity) and biomarkers for stress response and development (growth/appetite) were (i) examined across treatments and (ii) used to describe behavioral phenotypes via principal component analyses and general linear mixed models. Triploids exhibited a more active behavioral profile (p = 0.002), and those on a regular diet had greater Neuropeptide Y transcription (p = 0.02). A growth gene (early growth response protein 1, p = 0.02) and long-term neural development genes (neurogenic differentiation factor, p = 0.003 and synaptysomal-associated protein 25-a, p = 0.005) impacted activity and reactionary profiles, respectively. Overall, our probiotic treatment did not compensate for triploidy. Our research highlights novel applications of behavioral transcriptomics for identifying candidate genes and dynamic, mechanistic associations with complex behavioral repertoires.

The effect of Belin-Ambrósio Deviation (BAD-D) Index on two-year refractive outcomes of Photorefractive Keratectomy (PRK).

PURPOSE: To assess the effect of preoperative BAD-D index on two-year visual, refractive, and tomographic outcomes, as well as the efficacy and safety of PRK surgery. SETTING: Eye clinic, Mashhad, Iran. DESIGN: Prospective cohort study. METHODS: This study included 66 patients (66 eyes) who underwent PRK surgery, with a minimum follow-up period of 2 years. Participants were divided into two groups: preoperative BAD-D≥1.60 (high BAD-D) and preoperative BAD-D<1.60 (low BAD-D). Pre- and postoperative visual, refractive, and tomographic parameters were evaluated, and the efficacy and safety of the procedure were compared between groups. RESULTS: Sixty-six patients with a mean age of 35.50 years±8.21 (range 22 to 55 years) were included. Post-operatively, the mean spherical equivalent (SE) of refractive error was +0.32±0.65D in the high BAD-D group and +0.18±0.66D in the low BAD-D group (p=0.40). Also, at two years of follow-up, the mean uncorrected visual acuity was 0.98±0.07 in high BAD-D group and 0.97±0.08 in low BAD-D group (p=0.905). Among the postoperative tomographic parameters, front elevation thickness (F.EIe.Th), maximum Ambrósio relational thickness (ARTmax), astigmatism, and central corneal thickness were significantly different between the two groups (all, p< 0.05). At the two-year follow-up, the mean safety Index was 1.02±0.04, and 1.01±0.04 in high and low BAD-D groups respectively (p=0.37) and the mean efficacy Index was 0.99±0.07 and 0.98±0.06 in high and low BAD-D groups respectively (p=0.40). CONCLUSION: The preoperative BAD-D index does not predict postoperative visual, refractive, and tomographic outcomes in patients with low-to-moderate myopia. However, in patients with normal preoperative BAD-D values, higher agreement was expected between the attempted and achieved SE.