Pre-operative predicting factor in visual outcome after macular hole surgery.

OBJECTIVE: To determine the effectiveness of macular hole index (MHI) as a predicting factor of visual outcome after full thickness macular hole surgery. METHODS: This quasi-experimental study was conducted at LRBT Free Base Eye Hospital, Karachi from January 2018 to March 2019. Total 45 eyes of 45 patients with full thickness macular hole (FTMH) underwent preoperative Best Corrected Visual Acuity (BCVA) assessment with logMar chart and Optical Coherence Tomography (OCT) scanning, with measurement of base diameter and macular hole height. Values were calculated for the macular hole index (MHI), which was taken as the predictive factor. All patients had undergone 25+G trans-conjunctival three ports pars plana vitrectomy, internal limiting membrane peeling, and endo-tamponade of C3F8(14%). The final visual outcome of all the patients was noted. RESULTS: Forty-five patients were included for the study, out of which 10 (22.2%) were male and 35 (77.7%) were female. Age ranged from 45-70 years (mean age 57.20±6.47 years). The mean pre-operative visual acuity was 2.46±1.15 logMar and was 3.88±2.00 logMar, post-operatively. Moreover, 27(60.0%) out of 45 patients achieved BCVA (gain of 2 lines of the logMar chart). The average macular hole index was 1.55±0.50 and out of 45, 25 patients had MHI ≥0.5. It was found that patients with macular hole index ≥0.50 showed clinically significant improvement in BCVA in comparison to those who have macular hole index <0.50. CONCLUSION: Macular hole index can be used to predict functional success in macular hole surgery.

Faecalibacterium prausnitzii Strain HTF-F and Its Extracellular Polymeric Matrix Attenuate Clinical Parameters in DSS-Induced Colitis.

A decrease in the abundance and biodiversity of intestinal bacteria within the Firmicutes phylum has been associated with inflammatory bowel disease (IBD). In particular, the anti-inflammatory bacterium Faecalibacterium prausnitzii, member of the Firmicutes phylum and one of the most abundant species in healthy human colon, is underrepresented in the microbiota of IBD patients. The aim of this study was to investigate the immunomodulatory properties of F. prausnitzii strain A2-165, the biofilm forming strain HTF-F and the extracellular polymeric matrix (EPM) isolated from strain HTF-F. For this purpose, the immunomodulatory properties of the F. prausnitzii strains and the EPM were studied in vitro using human monocyte-derived dendritic cells. Then, the capacity of the F. prausnitzii strains and the EPM of HTF-F to suppress inflammation was assessed in vivo in the mouse dextran sodium sulphate (DSS) colitis model. The F. prausnitzii strains and the EPM had anti-inflammatory effects on the clinical parameters measured in the DSS model but with different efficacy. The immunomodulatory effects of the EPM were mediated through the TLR2-dependent modulation of IL-12 and IL-10 cytokine production in antigen presenting cells, suggesting that it contributes to the anti-inflammatory potency of F. prausnitzii HTF-F. The results show that F. prausnitzii HTF-F and its EPM may have a therapeutic use in IBD.

Functional metabolic map of Faecalibacterium prausnitzii, a beneficial human gut microbe.

The human gut microbiota plays a central role in human well-being and disease. In this study, we present an integrated, iterative approach of computational modeling, in vitro experiments, metabolomics, and genomic analysis to accelerate the identification of metabolic capabilities for poorly characterized (anaerobic) microorganisms. We demonstrate this approach for the beneficial human gut microbe Faecalibacterium prausnitzii strain A2-165. We generated an automated draft reconstruction, which we curated against the limited biochemical data. This reconstruction modeling was used to develop in silico and in vitro a chemically defined medium (CDM), which was validated experimentally. Subsequent metabolomic analysis of the spent medium for growth on CDM was performed. We refined our metabolic reconstruction according to in vitro observed metabolite consumption and secretion and propose improvements to the current genome annotation of F. prausnitzii A2-165. We then used the reconstruction to systematically characterize its metabolic properties. Novel carbon source utilization capabilities and inabilities were predicted based on metabolic modeling and validated experimentally. This study resulted in a functional metabolic map of F. prausnitzii, which is available for further applications. The presented workflow can be readily extended to other poorly characterized and uncharacterized organisms to yield novel biochemical insights about the target organism.

Antioxidants keep the potentially probiotic but highly oxygen-sensitive human gut bacterium Faecalibacterium prausnitzii alive at ambient air.

The beneficial human gut microbe Faecalibacterium prausnitzii is a 'probiotic of the future' since it produces high amounts of butyrate and anti-inflammatory compounds. However, this bacterium is highly oxygen-senstive, making it notoriously difficult to cultivate and preserve. This has so far precluded its clinical application in the treatment of patients with inflammatory bowel diseases. The present studies were therefore aimed at developing a strategy to keep F. prausnitzii alive at ambient air. Our previous research showed that F. prausnitzii can survive in moderately oxygenized environments like the gut mucosa by transfer of electrons to oxygen. For this purpose, the bacterium exploits extracellular antioxidants, such as riboflavin and cysteine, that are abundantly present in the gut. We therefore tested to what extent these antioxidants can sustain the viability of F. prausnitzii at ambient air. The present results show that cysteine can facilitate the survival of F. prausnitzii upon exposure to air, and that this effect is significantly enhanced the by addition of riboflavin and the cryoprotectant inulin. The highly oxygen-sensitive gut bacterium F. prausnitzii can be kept alive at ambient air for 24 h when formulated with the antioxidants cysteine and riboflavin plus the cryoprotectant inulin. Improved formulations were obtained by addition of the bulking agents corn starch and wheat bran. Our present findings pave the way towards the biomedical exploitation of F. prausnitzii in redox-based therapeutics for treatment of dysbiosis-related inflammatory disorders of the human gut.

How can Faecalibacterium prausnitzii employ riboflavin for extracellular electron transfer?

Faecalibacterium prausnitzii is one of the most abundant commensal microbes in the human gut. It is an important supplier of butyrate to the colonic epithelium, and low numbers of faecalibacteria have been associated with severe inflammatory bowel disease. Previous studies revealed that F. prausnitzii shuttles electrons extracellularly to oxygen in systems containing flavins and thiols. Since this electron shuttling to oxygen strongly stimulates growth, the present studies were aimed at elucidating the role of riboflavin as an extracellular electronophore of F. prausnitzii. We show that F. prausnitzii can use riboflavin as a mediator for extracellular electron transfer (EET) to the anode of microbial fuel cell systems. However, this bacterium relies on exogenous riboflavin, since it does not secrete this compound as shown by the analysis of a spent growth medium using cyclic voltammetry (CV). Importantly, CV showed that riboflavin can undergo fully reversible redox cycling under physiologically relevant conditions. Lastly, riboflavin is shown to mediate the electrochemical oxidation of the main bacterial reducing equivalent NADH. Based on our present observations, we hypothesize that riboflavin is of major importance as a redox mediator for bacterial EET and growth in the human gut.

The gut anaerobe Faecalibacterium prausnitzii uses an extracellular electron shuttle to grow at oxic-anoxic interphases.

Faecalibacterium prausnitzii is one of the most abundant bacteria in the human gut ecosystem and it is an important supplier of butyrate to the colonic epithelium. Low numbers of faecalibacteria have been associated with inflammatory bowel disease. Despite being extremely oxygen sensitive, F. prausnitzii is found adherent to the gut mucosa where oxygen diffuses from epithelial cells. This paradox is now explained on the basis of gas tube experiments, flavin-dependent reduction of 5,5'-dithiobis-2-nitrobenzoate and microbial fuel cell experiments. The results show that F. prausnitzii employs an extracellular electron shuttle of flavins and thiols to transfer electrons to oxygen. Both compounds are present in the healthy human gut. Our observations may have important implications for the treatment of patients with Crohn's disease, for example, with flavin- or antioxidant rich diets, and they provide a novel key insight in host-microbe interactions at the gut barrier.