Senait Assefa

Probiotic Interference of Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 with the Opportunistic Fungal Pathogen Candida albicans

Candida albicans is the most important Candida species causing vulvovaginal candidiasis (VVC). VVC has significant medical and economical impact on womens health and wellbeing. While current antifungal treatment is reasonably effective, supportive and preventive measures such as application of probiotics are required to reduce the incidence of VVC. We investigated the potential of the probiotics Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 towards control of C. albicans. In vitro experiments demonstrated that lactic acid at low pH plays a major role in suppressing fungal growth. Viability staining following cocultures with lactobacilli revealed that C. albicans cells lost metabolic activity and eventually were killed. Transcriptome analyses showed increased expression of stress-related genes and lower expression of genes involved in fluconazole resistance, which might explain the increased eradication of Candida in a previous clinical study on conjoint probiotic therapy. Our results provide insights on the impact of probiotics on C. albicans survival.

Status of bacterial colonization, Toll-like receptor expression and nuclear factor-kappa B activation in normal and diseased human livers.

Epidemiological data on bacterial translocation (BT), colonization and inflammation in normal human livers is lacking. In this study we investigated the status of bacterial colonization and inflammation in the normal, cirrhotic primary biliary cirrhosis (PBC), and nonalcoholic steatohepatitis (NASH) human liver tissues. Comparatively normal livers showed increased bacterial colonization than PBC and NASH. We analyzed mRNA levels of Toll-like receptors (TLR) 2 and TLR4, and protein levels of TLR4. Phosphorylated IKKα (pIKKα) protein estimation served as a marker for nuclear factor-kappa B (NF-κB) activation. In spite of the increased bacterial colonization in normal liver tissues, lower levels of TLR2/4 mRNA and TLR4 and pIKKα proteins were found compared to PBC and NASH indicating the maintenance of suppressed inflammation and immune tolerance in normal livers. To our knowledge, this is the first clinical evidence showing suppressed inflammation despite bacterial colonization in normal human livers thus maintaining liver immune homeostasis.

The effect of hyaluronic acid on biofunctionality of gelatin-collagen intestine tissue engineering scaffolds

The creation of engineered intestinal tissue has recently stimulated new endeavors with the ultimate goal of intestinal replacement for massive resections of bowel. In this context, we investigated the effect of hyaluronic acid (HA) on the physicochemical characteristics of gelatin-collagen scaf- folds and its cytocompatibilty to the human intestinal epithe- lial Caco-2 cell line in vitro. Gelatin/collagen hybrid scaffolds with different concentrations of HA were prepared by solvent casting and freeze-drying techniques and subsequent chemi- cal crosslinking by genipin. The morphologies of the scaffolds were characterized by scanning electron microscopy and Fou- rier transform infrared spectroscopy. In vitro tests were carried out in phosphate-buffered saline (PBS) solution to study the swelling ratio and the biostability of the scaffolds. It was found that the porous structure of the scaffolds could be tailored by further addition of HA. Moreover, both the swelling ratio and the degradation rate of the scaffold increased by addition of HA. A resazurin-based cell viability assay was employed to determine the viability and estimate the number of scaffold- adherent Caco-2 cells. The assay indicated that the scaffolds were all cytocompatible. We concluded that addition of less than 15% HA to scaffolds with a composition of 9:1 gelatin:col- lagen results only in incremental improvement in the struc- tural characteristics and cytocompatibility of the gelatin- collagen scaffolds. However, the scaffolds with 25% HA exhib- ited remarkable enhancement in physicochemical characteris- tics of the scaffolds including cell viability, growth, and attachment as well as their physical structure. V C 2013 Wiley

The effect of hyaluronic acid on biofunctionality of gelatin-collagen intestine tissue engineering scaffolds: Gelatin-Collagen Tissue Engineering Scaffolds

The creation of engineered intestinal tissue has recently stimulated new endeavors with the ultimate goal of intestinal replacement for massive resections of bowel. In this context, we investigated the effect of hyaluronic acid (HA) on the physicochemical characteristics of gelatin-collagen scaffolds and its cytocompatibilty to the human intestinal epithelial Caco-2 cell line in vitro. Gelatin/collagen hybrid scaffolds with different concentrations of HA were prepared by solvent casting and freeze-drying techniques and subsequent chemical crosslinking by genipin. The morphologies of the scaffolds were characterized by scanning electron microscopy and Fourier transform infrared spectroscopy. In vitro tests were carried out in phosphate-buffered saline (PBS) solution to study the swelling ratio and the biostability of the scaffolds. It was found that the porous structure of the scaffolds could be tailored by further addition of HA. Moreover, both the swelling ratio and the degradation rate of the scaffold increased by addition of HA. A resazurin-based cell viability assay was employed to determine the viability and estimate the number of scaffold-adherent Caco-2 cells. The assay indicated that the scaffolds were all cytocompatible. We concluded that addition of less than 15% HA to scaffolds with a composition of 9:1 gelatin:collagen results only in incremental improvement in the structural characteristics and cytocompatibility of the gelatin-collagen scaffolds. However, the scaffolds with 25% HA exhibited remarkable enhancement in physicochemical characteristics of the scaffolds including cell viability, growth, and attachment as well as their physical structure.

Lactobacilli with probiotic potential in the prairie vole (Microtus ochrogaster)

AbstractBackgroundRecent research suggests integration of the intestinal microbiota in gut-brain communication which could lead to new approaches to treat neurological disorders. The highly social prairie voles are an excellent model system to study the effects of environmental factors on social behavior. For future studies on the role of probiotics in ameliorating disorders with social withdrawal symptoms, we report the characterization of intestinal Lactobacillus isolates with probiotic potential from voles.Methods and results30 bacterial strains were isolated from the vole intestine and found to be distinct but closely related to Lactobacillus johnsonii using 16S rRNA gene sequencing and Random Amplification of Polymorphic DNA fingerprinting. In vitro characterizations including acid and bile tolerance, antimicrobial effects, antibiotic susceptibility, and adherence to intestinal epithelial cells were performed to assess the probiotic potential of selected strains. Since previous studies revealed that mercury ingestion triggers social deficits in voles, mercury resistance of the probiotic candidates was evaluated which could be an important factor in preventing/treating these behavioral changes.ConclusionsThis study demonstrates that lactobacilli with probiotic potential are present in the vole intestine. The Lactobacillus isolates identified in this study will provide a basis for the investigation of probiotic effects in the vole behavioral model system.

Fecal microbiota in the female prairie vole (Microtus ochrogaster)

We examined the fecal microbiota of female prairie voles. This species is socially and, likely, sexually monogamous, and thus serves as a valuable model in which to examine the interaction between the microbiota-gut-brain axis and social behavior. At present, little is known about the gastrointestinal microbiota of prairie voles; therefore, we performed a first characterization of the fecal microbiota using 16S rRNA gene amplicon sequencing. Semiconductor sequencing technology on an Ion Torrent PGM platform was used to assess the composition of fecal microbiotas from twelve female prairie voles. Following quality filtering, 1,017,756 sequencing reads were classified from phylum to genus level. At the phylum level, Firmicutes, Bacteroidetes, and Saccharibacteria were the predominant taxa, while the Bacteriodales, Erysipelotrichaceae, Ruminococcaceae, and Lachnospiraceae contributed the most dominant microbial groups and genera. Microbial community membership was most similar between vole sibling pairs, but consideration of taxon abundances weakened these associations. The interdependence of host factors such as genetics and behavior with the gastrointestinal microbiota is likely to be particularly pronounced in prairie voles. Our pilot characterization of the prairie vole intestinal microbiota revealed a microbial community composition remarkably consistent with the monogastric alimentary system of these rodents and their diet rich in complex plant carbohydrates. The highly social nature of these animals poses specific challenges to microbiome analyses that nonetheless are valuable for advancing research on the microbiota-gut-brain-behavior axis. Our study provides an important basis for future microbiome research in this emerging model organism for studying social behavior.