In vitro adhesion and anti-inflammatory properties of Limosilactobacillus fermentum FS-10 isolated from infant fecal sample / Adhesión in vitro y propiedades antiinflamatorias de Limosilactobacillus fermentum FS-10 aislado de muestra fecal infantil

In this study, seven strains of Limosilactobacillus fermentum were isolated from an infant fecal sample and characterized using in vitro studies. Lactobacillus rhamnosus GG was used as a comparison because it is a well-documented commercial probiotic. The isolates were tested for attributes such as acid and phenol tolerance, bile salt hydrolase (BSH) activity, and antibiotic sensitivity. One isolate, L. fermentum FS-10, displayed enhanced cell surface hydrophobicity (> 85%) and mucin adhesion. Mucin-binding helps colonization in the gut. The immunomodulatory property of L. fermentum FS-10 was evaluated by determining the modulation of pro- and anti-inflammatory factors such as tumor necrosis factor-alpha (TNF-α), interleukin (IL)-10, and nitric oxide (NO) in human acute monocytic leukemia (THP-1) cells under inflammatory conditions induced by lipopolysaccharide (LPS). L. fermentum FS-10 potently downregulated the expression of TNF-α and nitric oxide and upregulated IL-10 levels, indicating an anti-inflammatory response. Safety assessment of the strain revealed the absence of genes for virulence factors, toxin production, and antibiotic resistance, potentiating application as a probiotic strain.(AU)

In vitro adhesion and anti-inflammatory properties of Limosilactobacillus fermentum FS-10 isolated from infant fecal sample.

In this study, seven strains of Limosilactobacillus fermentum were isolated from an infant fecal sample and characterized using in vitro studies. Lactobacillus rhamnosus GG was used as a comparison because it is a well-documented commercial probiotic. The isolates were tested for attributes such as acid and phenol tolerance, bile salt hydrolase (BSH) activity, and antibiotic sensitivity. One isolate, L. fermentum FS-10, displayed enhanced cell surface hydrophobicity (> 85%) and mucin adhesion. Mucin-binding helps colonization in the gut. The immunomodulatory property of L. fermentum FS-10 was evaluated by determining the modulation of pro- and anti-inflammatory factors such as tumor necrosis factor-alpha (TNF-α), interleukin (IL)-10, and nitric oxide (NO) in human acute monocytic leukemia (THP-1) cells under inflammatory conditions induced by lipopolysaccharide (LPS). L. fermentum FS-10 potently downregulated the expression of TNF-α and nitric oxide and upregulated IL-10 levels, indicating an anti-inflammatory response. Safety assessment of the strain revealed the absence of genes for virulence factors, toxin production, and antibiotic resistance, potentiating application as a probiotic strain.

Retinal Hemangioblastoma with Extraocular Extension: Report of Three Cases.

Retinal hemangioblastoma (RH) is the earliest and most common clinical manifestation in Von Hippel-Lindau (VHL) disease. RH can also present in isolation without any evidence of VHL. Clinical course of RH can be stationary or progress to exudation and chronic retinal detachment requiring surgical intervention. We report 3 cases of aggressive RH with extraocular extension in young males causing painful blind eye requiring enucleation. Two of our cases were bilateral involvement and had systemic manifestations of VHL. The third patient had unilateral involvement with no systemic evidence of VHL. This manifestation of RH is rarely reported. Two of our patients with VHL had early manifestations of RH and had undergone multiple cryotherapy sessions as well as retinal detachment surgery for exudative retinal detachment. This differential should be considered in vascular lesion arising from intraocular structures especially in diagnosed patients of VHL. The cases also highlight the aggressive behaviour and long-term progression of RH in some patients despite early treatment.

Acute glycemic and insulin response of Fossence™ alone, or when substituted or added to a carbohydrate challenge: A three-phase, acute, randomized, cross-over, double blind clinical trial.

Short chain fructo-oligosaccharides (scFOS) are well-recognized prebiotic fibers. Fossence™ (FOSS) is a scFOS that has been produced from sucrose via a proprietary fermentation process and has not been tested for its digestibility or glucose/insulin response (GR and IR, respectively). The present randomized, controlled, cross-over study was conducted in 3 phases to explore GR and IR to ingestion of FOSS, when replaced by/added to available-carbohydrates (avCHO) among 25 healthy adults (40 ± 14years). In each phase GR and IR elicited by 3-4 test-meals were measured among the fasted recruited subjects. The interventional test meals were as follows: Phase-1, water alone or 10g FOSS or 10g Dextrose in 250ml water; Phase-2, 250ml water containing Dextrose:FOSS (g:g) in the content as 50:0 or 50:15 or 35:0 or 35:15; Phase-3 portions of white-bread (WB) containing avCHO:FOSS (g:g) in the content as 50:0 or 50:15 or 35:0 or 35:15. Blood samples (finger prick method) were collected at fasting and 15, 30, 45, 60, 90 and 120 min after start of test meal ingestion. Plasma glucose and serum insulin were analyzed utilizing standard methods. The primary endpoint was differences in glucose IAUC. All subjects provided their written consent to participate in the study (ClinicalTrials.gov: NCT03755232). The results demonstrated that FOSS, when consumed alone, showed no raise in glycaemia or insulinemia and was statistically equivalent to response of water alone. GR and IR elicited by dextrose:FOSS and WB:FOSS test-meals of Phase 2 and Phase 3, were statistically equivalent to the respective test-meals without FOSS. Result of the 3 phases support the hypothesis that FOSS is resistant to breakdown and is indigestible in the human small-intestine, and therefore can be classified as an unavailable carbohydrate that does not raise post prandial blood glucose or insulin. FOSS, being sweet to taste, may be an acceptable sugar replacer in beverages without compromising their taste and sensory qualities.

Malignant tumors of the eyelid in India: A multicenter, multizone study on clinicopathologic features and outcomes.

PURPOSE: To analyze data on eyelid malignancy in India, clinical and pathologic features, and outcomes. METHODS: A multicentre study, from oculoplastic practices in four geographic zones in India. The centers perform similar documentation and protocol-based management for eyelid tumors. Clinical features, pathology, American Joint Committee on Cancer (AJCC) class, management, and the outcomes were analyzed. RESULTS: The study included 129 patients, with slight female preponderance and mean age 62.7 years. The median delay to the consultation was 9 months. Rural patients traveled a mean distance of 115.2 km; there was no difference between the city and outstation patients in the delay to consultation or follow up. Pathology included 55/129 (42.6%) sebaceous gland carcinoma (SGC), 47/129 (36.4%) basal cell carcinoma (BCC), squamous cell carcinoma (SCC) in 15 (11.6%), and 12 (9.3%) other tumors. Commonest AJCC class was T2b/T3a in 80/111 (72%), invasion of the orbit was present in 16 (12.4%). Surgery with margin clearance was performed in 103. With a mean follow-up of 21.44 months, local recurrence and/or metastasis were seen in 12%. The diagnosis of SGC was strongly associated with adverse outcomes (odds ratio: 7.36). On multiple logistic regression analysis, diagnosis of SGC (P = 0.011) was significant in having adverse outcomes. CONCLUSION: The multicenter Indian data shows the highest prevalence of SGC, with the commonest AJCC class T2b. Most tumors were locally resectable at presentation. The histopathologic diagnosis of SGC is the factor strongly associated with adverse outcomes.

Provitamin D3 modulation through prebiotics supplementation: simulation based assessment.

Vitamin D is important in multiple health conditions. Vitamin D deficiency is prevalent globally even with exposure to adequate sunlight. Reduction in provitamin D3 (7-dehydrocholesterol, 7-DHC) is an important cause of vitamin D3 deficiency. Vitamin supplementation, food fortification, and use of probiotics are some approaches to reduce vitamin D3 deficiency. This study investigates plausibility of 7-DHC biosynthesis through dietary prebiotics supplementation. Furthermore, it reports mechanistic details and constraints for the biosynthesis using flux balance analysis (FBA) simulations. The FBA simulations using co-metabolism models comprising human host and a resident bacterium (Faecalibacterium prausnitzii or Bacteroides thetaiotamicron) indicated increased flux of 7-DHC with short-chain fructooligosaccharide (scFOS) or inulin supplementation. We observed around 2-fold increase in flux compared to the baseline. Biosynthesis of 7-DHC was primarily modulated through acetate, pyruvate and lactate secreted by the bacterium. We observed diverse mechanisms and dose dependent responses. We extended this assessment to 119 resident bacteria and investigated the metabolites profiles with prebiotics supplementation. In summary, the current study suggests the potential use of applying prebiotics in enhancing 7-DHC biosynthesis. Furthermore, performance of the different gut bacteria with prebiotic supplementation for secreted metabolites profile is reported. These results may be useful to design future clinical studies.

Biosynthesis of Hyaluronic acid polymer: Dissecting the role of sub structural elements of hyaluronan synthase.

Hyaluronic acid (HA) based biomaterials have several biomedical applications. HA biosynthesis is catalysed by hyaluronan synthase (HAS). The unavailability of 3-D structure of HAS and gaps in molecular understanding of HA biosynthesis process pose challenges in rational engineering of HAS to control HA molecular weight and titer. Using in-silico approaches integrated with mutation studies, we define a dictionary of sub-structural elements (SSE) of the Class I Streptococcal HAS (SeHAS) to guide rational engineering. Our study identifies 9 SSE in HAS and elucidates their role in substrate and polymer binding and polymer biosynthesis. Molecular modelling and docking assessment indicate a single binding site for two UDP-substrates implying conformationally-driven alternating substrate specificities for this class of enzymes. This is the first report hypothesizing the involvement of sites from SSE5 in polymer binding. Mutation at these sites influence HA production, indicating a tight coupling of polymer binding and synthase functions. Mutation studies show dispensable role of Lys-139 in substrate binding and a key role of Gln-248 and Thr-283 in HA biosynthesis. Based on the functional architecture in SeHAS, we propose a plausible three-step polymer extension model from its reducing end. Together, these results open new avenues for rational engineering of Class I HAS to study and regulate its functional properties and enhanced understanding of glycosyltransferases and processive enzymes.

The ocular surface after simple limbal epithelial transplant (SLET): A high-resolution OCT study of the early postoperative period.

Simple limbal epithelial transplantation (SLET) is an emerging technique for treating unilateral limbal stem cell deficiency. We report the high-resolution, anterior segment optical coherence tomography (OCT) features of the first 2 weeks of a patient undergoing SLET for an old acid injury of the right eye, repeatedly from postoperative day 1 through day 14. Three out of 11 explants with the subjacent human amniotic membrane (hAM) and the overlaid bandage contact lens were imaged. The hAM was intact and of the same thickness throughout the study period; the sub-hAM space increased from day 3 to 9 and disappeared by day 10; the explants started thinning from day 3 with the fibrin around them starting to decrease from day 2 and completely disappeared by day 4. Epithelialization occurred between day 8 and 14 and proceeded more rapidly towards the limbus than centrally. There was no change of the corneal stromal thickness or reflectivity. This case report uses high-definition, spectral-domain OCT to document the events on the ocular surface after a successful SLET surgery and opens up an avenue to study epithelialization in a convenient and noninvasive manner.

A prospective randomized, double-blind, placebo-controlled, dose-response relationship study to investigate efficacy of fructo-oligosaccharides (FOS) on human gut microflora.

Fructo-oligosaccharides (FOS), a prebiotic supplement, is known for its Bifidogenic capabilities. However, aspects such as effect of variable quantities of FOS intake on gut microbiota, and temporal dynamics of gut microbiota (transitioning through basal, dosage, and follow-up phases) has not been studied in detail. This study investigated these aspects through a randomized, double-blind, placebo-controlled, dose-response relationship study. The study involved 80 participants being administered FOS at three dose levels (2.5, 5, and 10 g/day) or placebo (Maltodextrin 10 g/day) during dosage phase. Microbial DNA extracted from fecal samples collected at 9 intervening time-points was sequenced and analysed. Results indicate that FOS consumption increased the relative abundance of OTUs belonging to Bifidobacterium and Lactobacillus. Interestingly, higher FOS dosage appears to promote, in contrast to Maltodextrin, the selective proliferation of OTUs belonging to Lactobacillus. While consumption of prebiotics increased bacterial diversity, withdrawal led to its reduction. Apart from probiotic bacteria, a significant change was also observed in certain butyrate-producing microbes like Faecalibacterium, Ruminococcus and Oscillospira. The positive impact of FOS on butyrate-producing bacteria and FOS-mediated increased bacterial diversity reinforces the role of prebiotics in conferring beneficial functions to the host.

LogMPIE, pan-India profiling of the human gut microbiome using 16S rRNA sequencing.

The "Landscape Of Gut Microbiome - Pan-India Exploration", or LogMPIE study, is the first large-scale, nationwide record of the Indian gut microbiome. The primary objective of the study was to identify and map the Indian gut microbiome baseline. This observational study was conducted across 14 geographical locations in India. Enrolled subjects were uniformly distributed across geographies (north, east, west and south) and body mass index (obese and non-obese). Furthermore, factors influencing the microbiome, such as age and physical activity, were also considered in the study design. The LogMPIE study recorded data from 1004 eligible subjects and reported 993 unique microorganisms across the Indian microbiome diaspora. The data not only map the Indian gut microbiome baseline but also function as a useful resource to study, analyse and identify signatures characterizing the physiological dispositions of the subjects. Furthermore, they provide insight into the unique features describing the Indian microbiome. The data are open and may be accessed from the European Nucleotide Archive (ENA) portal of the European Bioinformatics Institute (https://www.ebi.ac.uk/ena/data/view/PRJEB25642).

Primary limbal stem cell transplantation in the surgical management of extensive ocular surface squamous neoplasia involving the limbus.

PURPOSE: To determine the role of primary limbal stem cell transplantation (LSCT) in managing extensive ocular surface squamous neoplasia (OSSN) with more than 3 quadrants of limbal involvement to prevent manifestations of limbal stem cell deficiency. METHODS: A multi-center, comparative, and interventional clinical study of cases and historical controls was done. Patients with a limbal mass suggestive of OSSN involving more than 3 quadrants of limbus (> 9 clock hours) and no local, regional or systemic metastasis, who underwent primary surgical excision with or without adjuvant chemotherapy and had a minimum follow-up of 12 months were included in this study. Two historical controls underwent tumor excision with only amniotic membrane grafting. Three cases underwent tumor excision along with LSCT in the form of conjunctival limbal autograft (CLAG) or simple limbal epithelial transplant (SLET). RESULTS: None of the eyes had any recurrences of OSSN during the follow-up period. Cases with primary LSCT with tumor excision showed better ocular surface stability in the long term. CONCLUSION: Concomitant LSCT either in the form of CLAG or SLET appears to provide better long-term outcomes and is advocated while excision of extensive OSSN involving the limbus.

SimCAL: a flexible tool to compute biochemical reaction similarity.

BACKGROUND: Computation of reaction similarity is a pre-requisite for several bioinformatics applications including enzyme identification for specific biochemical reactions, enzyme classification and mining for specific inhibitors. Reaction similarity is often assessed at either two levels: (i) comparison across all the constituent substrates and products of a reaction, reaction level similarity, (ii) comparison at the transformation center with various degrees of neighborhood, transformation level similarity. Existing reaction similarity computation tools are designed for specific applications and use different features and similarity measures. A single system integrating these diverse features enables comparison of the impact of different molecular properties on similarity score computation. RESULTS: To address these requirements, we present SimCAL, an integrated system to calculate reaction similarity with novel features and capability to perform comparative assessment. SimCAL provides reaction similarity computation at both whole reaction level and transformation level. Novel physicochemical features such as stereochemistry, mass, volume and charge are included in computing reaction fingerprint. Users can choose from four different fingerprint types and nine molecular similarity measures. Further, a comparative assessment of these features is also enabled. The performance of SimCAL is assessed on 3,688,122 reaction pairs with Enzyme Commission (EC) number from MetaCyc and achieved an area under the curve (AUC) of > 0.9. In addition, SimCAL results showed strong correlation with state-of-the-art EC-BLAST and molecular signature based reaction similarity methods. CONCLUSIONS: SimCAL is developed in java and is available as a standalone tool, with intuitive, user-friendly graphical interface and also as a console application. With its customizable feature selection and similarity calculations, it is expected to cater a wide audience interested in studying and analyzing biochemical reactions and metabolic networks.

ReactPRED: a tool to predict and analyze biochemical reactions.

MOTIVATION: Biochemical pathways engineering is often used to synthesize or degrade target chemicals. In silico screening of the biochemical transformation space allows predicting feasible reactions, constituting these pathways. Current enabling tools are customized to predict reactions based on pre-defined biochemical transformations or reaction rule sets. Reaction rule sets are usually curated manually and tailored to specific applications. They are not exhaustive. In addition, current systems are incapable of regulating and refining data with an aim to tune specificity and sensitivity. A robust and flexible tool that allows automated reaction rule set creation along with regulated pathway prediction and analyses is a need. ReactPRED aims to address the same. RESULTS: ReactPRED is an open source flexible and customizable tool enabling users to predict biochemical reactions and pathways. The tool allows automated reaction rule creation from a user defined reaction set. Additionally, reaction rule degree and rule tolerance features allow refinement of predicted data. It is available as a flexible graphical user interface and a console application. AVAILABILITY AND IMPLEMENTATION: ReactPRED is available at: https://sourceforge.net/projects/reactpred/ CONTACT: anirban.b@samsung.com or ty76.kim@samsung.comSupplementary information: Supplementary data are available at Bioinformatics online.

Quantitative Assessment of Heart Rate Dynamics during Meditation: An ECG Based Study with Multi-Fractality and Visibility Graph.

The cardiac dynamics during meditation is explored quantitatively with two chaos-based non-linear techniques viz. multi-fractal detrended fluctuation analysis and visibility network analysis techniques. The data used are the instantaneous heart rate (in beats/minute) of subjects performing Kundalini Yoga and Chi meditation from PhysioNet. The results show consistent differences between the quantitative parameters obtained by both the analysis techniques. This indicates an interesting phenomenon of change in the complexity of the cardiac dynamics during meditation supported with quantitative parameters. The results also produce a preliminary evidence that these techniques can be used as a measure of physiological impact on subjects performing meditation.

RxnSim: a tool to compare biochemical reactions.

UNLABELLED: : Quantitative assessment of chemical reaction similarity aids database searches, classification of reactions and identification of candidate enzymes. Most methods evaluate reaction similarity based on chemical transformation patterns. We describe a tool, RxnSim, which computes reaction similarity based on the molecular signatures of participating molecules. The tool is able to compare reactions based on similarities of substrates and products in addition to their transformation. It allows masking of user-defined chemical moieties for weighted similarity computations. AVAILABILITY AND IMPLEMENTATION: RxnSim is implemented in R and is freely available from the Comprehensive R Archive Network, CRAN (http://cran.r-project.org/web/packages/RxnSim/). CONTACT: anirban.b@samsung.com or ty76.kim@samsung.com SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Enhancement of the substrate scope of transketolase.

To enhance the activity of transketolase towards nonphosphorylated substrates and enlarge the scope of its substrates, notably to long polyol aldehyde acceptors (D-ribose or D-glucose), a rational design-supported evolution strategy was applied. By using docking experiments, an in silico library, and iterative mutagenesis, libraries of single- and double-point mutants were designed and generated. A double-screening approach was implemented, coupling a preselection activity assay (HPLC method) and a selective assay (GC method) to find the best enzymes. Several mutants (R526N, R526Q, R526Q/S525T, R526K/S525T) showed improved activities towards nonphosphorylated substrates as the coupled products of lithium hydroxypyruvate (HPA) with glycolaldehyde (GO), D-ribose or D-glucose. These mutated enzymes were further characterised. They were shown to be up to four times more active than the wild-type (mutant R526Q/S525T) for nonphosphorylated substrates LiHPA/GO (V(m) /K(m) for LiHPA = 92.4 instead of 28.8×10(-3) min(-1) for the wild-type) and 2.6 times more active for substrates LiHPA/rib.

Custom ocular prosthesis in children: how often is a change required?

PURPOSE: To evaluate the indications, outcome and replacement schedule of custom ocular prostheses (COP) in children. METHODS: Retrospective review evaluating indications, outcome and replacement schedule of COP in children (≤16 years). RESULTS: Three hundred and thirty (330) children were fitted with COP. With average follow-up of 18.05 months (range 1-50 months), 136(41.2%) children needed replacement/modification of the prosthesis. Information regarding indications for replacement/modification of prosthesis was available in 125 cases. Indications included enophthalmic prosthesis in 34(27.2%), prosthesis rotation within the socket in 25(20%), loose fit in 16(12.8%), decentration of the cornea in 15(12%), color touch up in 13(10.4%), lost prosthesis in 8(6.4%), cosmetically significant ptosis in 2(1.6%), replacement following implant exchange surgery in 2(1.6%) and combination of above factors in 10(8%). Time for replacement/modification of prosthesis was assessed in 3 age groups. In ≤3 years, 47% (36 out of 76) children underwent replacement/modification at a mean duration of 18 months (range 3-39 months) from the date of prosthesis fitting. In the 3-12 years age group, 43% (80 out of 184) underwent replacement/modification at 21 months (range 2-48 months); and in the 12-16 year age group, 29% (20 out of 70) underwent replacement/modification of prosthesis at a mean duration of 26 months (range 3-50 months). CONCLUSION: A change of prosthesis is required between 18-26 months following prosthesis placement in children. Over an average of 18 months, a change in the prosthesis was required in 41%, with the youngest age group having the highest exchange rate (47%), and the oldest group the lowest (29%).

Centurion syndrome: clinical presentation and surgical outcome.

PURPOSE: To describe the clinical signs and outcome of surgery in cases with Centurion syndrome and highlight the clinical presentation of this syndrome. METHODS: A retrospective review of records of patients with Centurion syndrome who underwent surgery between January 2002 and July 2007. The position of the punctum, nasal bridge prominence, Hertel exophthalmometry and the presence of inferiorly directed sharp angulation of the medial canthus was noted. The patients underwent anterior canthal tendon release, punctoplasty or medial conjunctivoplasty or a combination of these. RESULTS: Of the 13 cases, 10 had been treated medically elsewhere. The mean age of onset was 20.7 years. Most of the patients were male 10 (76.9%). A prominent nasal bridge was demonstrable in 9 (69.2%) patients and sharp inferior angulation of the medial canthus, the 'beak' sign was present in 9 (69.2%) cases. Anterior canthal tendon release alone was performed in 4 patients, in combination with punctoplasty in 5 and with conjunctivoplasty in 4. Watering resolved in 10 patients. 3 patients who had a combination of anterior canthal tendon release and punctoplasty had persistent but asymptomatic epiphora. DISCUSSION: Centurion syndrome is characterised by the abnormal anterior insertion of the medial canthal tendon with displacement of the punctum out of the lacrimal lake. Most cases present with unexplained watering. Surgical management by anterior canthal tendon release with or without conjunctivoplasty or lower lid retractor plication is usually successful.