scholarly journals Pre-Growth Culture Conditions Affect Type 1 Fimbriae-Dependent Adhesion of Salmonella

2020 ◽  
Vol 21 (12) ◽  
pp. 4206 ◽  
Author(s):  
Beata Klasa ◽  
Anna Ewa Kędzierska ◽  
Krzysztof Grzymajło
Keyword(s):  
Growth Conditions ◽  
Culture Conditions ◽  
Infection Process ◽  
Intestinal Epithelial ◽  
Exact Role ◽  
Type 1 Fimbriae ◽  
Wide Range ◽  
Host Cell Interactions ◽  
Host Tissues

Among various fimbrial structures used by Salmonella enterica to colonize host tissues, type 1 fimbriae (T1F) are among the most extensively studied. Although some experiments have shown the importance of T1F in the initial stages of Salmonella infection, their exact role in the infection process is not fully known. We suggested that different outcomes of T1F investigations were due to the use of different pre-infection growth conditions for the induction of the T1F. We utilized qPCR, flow cytometry, and a wide range of adhesion assays to investigate Salmonella Choleraesuis and Salmonella Typhimurium adhesion in the context of T1F expression. We demonstrated that T1F expression was highly dependent on the pre-infection growth conditions. These growth conditions yielded T1F+ and T1F- populations of Salmonella and, therefore, could be a factor influencing Salmonella-host cell interactions. We supported this conclusion by showing that increased levels of T1F expression directly correlated with higher levels of Salmonella adherence to the intestinal epithelial IPEC-J2 cell line.

scholarly journals HilE is required for synergistic activation of SPI-1 gene expression in Salmonella enterica serovar Typhimurium

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Selwan Hamed ◽  
Riham M. Shawky ◽  
Mohamed Emara ◽  
James M. Slauch ◽  
Christopher V. Rao
Keyword(s):  
Gene Expression ◽  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Growth Conditions ◽  
Intestinal Epithelial ◽  
Synergistic Activation ◽  
Wide Range ◽  
Serovar Typhimurium ◽  
In The Wild ◽  
Intestinal Pathogen

Abstract Background Salmonella enterica serovar Typhimurium is an intestinal pathogen capable of infecting a wide range of animals. It initiates infection by invading intestinal epithelial cells using a type III secretion system encoded within Salmonella pathogenicity island 1 (SPI-1). The SPI-1 genes are regulated by multiple interacting transcription factors. The master regulator is HilD. HilE represses SPI-1 gene expression by binding HilD and preventing it from activating its target promoters. Previous work found that acetate and nutrients synergistically induce SPI-1 gene expression. In the present study, we investigated the role of HilE, nominally a repressor of SPI-1 gene expression, in mediating this response to acetate and nutrients. Results HilE is necessary for activation of SPI-1 gene expression by acetate and nutrients. In mutants lacking hilE, acetate and nutrients no longer increase SPI-1 gene expression but rather repress it. This puzzling response is not due to the BarA/SirA two component system, which governs the response to acetate. To identify the mechanism, we profiled gene expression using RNAseq in the wild type and a ΔhilE mutant under different growth conditions. Analysis of these data suggested that the Rcs system, which regulates gene expression in response to envelope stress, is involved. Consistent with this hypothesis, acetate and nutrients were able to induce SPI-1 gene expression in mutants lacking hilE and the Rcs system. Conclusions While the exact mechanism is unknown, these results demonstrate the HilE, nominally a repressor of SPI-1 gene expression, can also function as an activator under the growth conditions investigated. Collectively, these results provide new insights regarding SPI-1 gene regulation and demonstrate that HilE is more complex than initially envisioned.

scholarly journals Over 2000-fold increased production of the leaderless bacteriocin garvicin KS by genetic engineering and optimization of culture conditions

2018 ◽  
Author(s):  
Amar A. Telke ◽  
Kirill V. Ovchinnikov ◽  
Kiira S. Vuoristo ◽  
Geir Mathiesen ◽  
Tage Thorstensen ◽  
...  
Keyword(s):  
Growth Conditions ◽  
Culture Conditions ◽  
Research Field ◽  
Bacteriocin Production ◽  
Gene Dose ◽  
Constant Ph ◽  
Wide Range ◽  
Bottle Neck ◽  
High Production ◽  
The Impact

AbstractThe leaderless bacteriocin Garvicin KS (GarKS) is a potent antimicrobial, being active against a wide range of important pathogens. GarKS production by the native producerLactococcus garvieaeKS1546 was however relatively low (80 BU/ml) under standard laboratory growth conditions (batch culture in GM17 at 30°C). To improve the production of GarKS, we systematically evaluated the impact of different media and media components on bacteriocin production. Based on the outcomes a new medium formulation was made to greatly improve bacteriocin production. The new medium composed of pasteurized milk and tryptone (PM-T), increased GarKS production about 60-fold compared to that achieved in GM17. GarKS production was increased further 4-fold (i.e., to 20,000 BU/ml) by increasing gene dose of the bacteriocin gene cluster (gak) in the native producer. Finally, a combination of the newly composed medium (PM-T), an increased gene dose and a cultivation at a constant pH 6 and a 50-60% dissolved oxygen level in growth medium, gave rise to a GarKS production of 164,000 BU/ml. This high production, which is about 2000-fold higher compared to that initially achieved in GM17, corresponds to a GarKS production of 1.2 g/L. To our knowledge, this is one of the highest bacteriocin production reported hitherto.ImportanceLow bacteriocin production is a well-known bottle-neck in developing bacteriocins into large-scaled and useful applications. The present study shows different approaches that significantly improve bacteriocin production. This is an important research field to better exploit the antimicrobial potential of bacteriocins, especially with regard to the decreasing effect of antibiotics in infection treatments due to the global emergence of antibiotic resistance.

scholarly journals Type 1 Fimbriation and Phase Switching in a NaturalEscherichia coli fimB Null Strain, Nissle 1917

1999 ◽  
Vol 181 (24) ◽  
pp. 7470-7478 ◽  
Author(s):  
Bodil Stentebjerg-Olesen ◽  
Trinad Chakraborty ◽  
Per Klemm
Keyword(s):  
Low Frequency ◽  
Growth Conditions ◽  
Current Evidence ◽  
Phase Variable ◽  
Insertion Element ◽  
Phase Switching ◽  
Type 1 Fimbriae ◽  
Tyrosine Recombinases ◽  
Static Conditions

ABSTRACT Escherichia coli Nissle 1917 has been used as a probiotic against intestinal disorders for many decades. It is a good colonizer of the human gut and has been reported to be able to express type 1 fimbriae. Type 1 fimbriae are surface organelles which mediate α-d-mannose-sensitive binding to various host cell surfaces. The expression is phase variable, and two tyrosine recombinases, FimB and FimE, mediate the inversion of the fimbrial phase switch. Current evidence suggests that FimB can carry out recombination in both directions, whereas FimE-catalyzed switching is on to off only. We show here that under liquid shaking growth conditions, Nissle 1917 did not express type 1 fimbriae, due to a truncation of the fimB gene by an 1,885-bp insertion element. Despite its fimB null status, Nissle 1917 was still capable of off-to-on switching of the phase switch and expressing type 1 fimbriae when grown under static conditions. This phase switching was not catalyzed by FimE, by truncated FimB, or by information residing within the insertion element. No further copies offimB seemed to be present on the chromosome of Nissle 1917, suggesting that another tyrosine recombinase in Nissle 1917 is responsible for the low-frequency off-to-on inversion of the phase switch that is strongly favored under static growth conditions. This is the first report documenting the non-FimB- or non-FimE-catalyzed inversion of the fim switch.

scholarly journals UropathogenicEscherichia coliExpress Type 1 Fimbriae Only in Surface Adherent Populations Under Physiological Growth Conditions

2015 ◽  
Vol 213 (3) ◽  
pp. 386-394 ◽  
Author(s):  
Kristian Stærk ◽  
Surabhi Khandige ◽  
Hans Jørn Kolmos ◽  
Jakob Møller-Jensen ◽  
Thomas Emil Andersen
Keyword(s):  
Growth Conditions ◽  
Type 1 Fimbriae

Glycans in scaffold design in tissue reconstruction

2021 ◽  
pp. 088391152199784
Author(s):  
Nipun Jain ◽  
Shashi Singh
Keyword(s):  
Cell Attachment ◽  
Low Cost ◽  
Growth Conditions ◽  
Downstream Signaling ◽  
Cell Carrier ◽  
Wide Range ◽  
Proliferation And Differentiation ◽  
Different Types ◽  
Tissue Reconstruction ◽  
A Cell

Development of an artificial tissue by tissue engineering is witnessed to be one of the long lasting clarified solutions for the damaged tissue function restoration. To accomplish this, a scaffold is designed as a cell carrier in which the extracellular matrix (ECM) performs a prominent task of controlling the inoculated cell’s destiny. ECM composition, topography and mechanical properties lead to different types of interactions between cells and ECM components that trigger an assortment of cellular reactions via diverse sensing mechanisms and downstream signaling pathways. The polysaccharides in the form of proteoglycans and glycoproteins yield better outcomes when included in the designed matrices. Glycosaminoglycan (GAG) chains present on proteoglycans show a wide range of operations such as sequestering of critical effector morphogens which encourage proficient nutrient contribution toward the growing stem cells for their development and endurance. In this review we discuss how the glycosylation aspects are of considerable importance in everyday housekeeping functions of a cell especially when placed in a controlled environment under ideal growth conditions. Hydrogels made from these GAG chains have been used extensively as a resorbable material that mimics the natural ECM functions for an efficient control over cell attachment, permeability, viability, proliferation, and differentiation processes. Also the incorporation of non-mammalian polysaccharides can elicit specific receptor responses which authorize the creation of numerous vigorous frameworks while prolonging the low cost and immunogenicity of the substance.

AGN1 vs AGN2 dichotomy as seen from the point of view of ionized outflows

2019 ◽  
Vol 15 (S356) ◽  
pp. 96-96
Author(s):  
Eleonora Sani
Keyword(s):  
Accretion Rate ◽  
Point Of View ◽  
Emission Lines ◽  
Black Hole Mass ◽  
X Ray ◽  
Bolometric Luminosity ◽  
Wide Range ◽  
Strong Contrast

AbstractI present a detailed study of ionized outflows in a large sample of 650 hard X-ray detected AGN. Taking advantage of the legacy value of the BAT AGN Spectroscopic Survey (BASS, DR1), we are able to reveal the faintest wings of the [OIII] emission lines associated with outflows. The sample allows us to derive the incidence of outflows covering a wide range of AGN bolometric luminosity and test how the outflow parameters are related with various AGN power tracers, such as black hole mass, Eddington ratio, luminosity. I’ll show how ionized outflows are more frequently found in type 1.9 and type 1 AGN (50% and 40%) with respect to the low fraction in type 2 AGN (20%). Within such a framework, I’ll demonstrate how type 2 AGN outflows are almost evenly balanced between blue- and red-shifted winds. This, in strong contrast with type 1 and type 1.9 AGN outflows which are almost exclusively blue-shifted. Finally, I’ll prove how the outflow occurrence is driven by the accretion rate, whereas the dependence of outflow properties with respect to the other AGN power tracers happens to be quite mild.

scholarly journals A global perspective on the issue of access to insulin

Diabetologia ◽  
2021 ◽  
Author(s):  
David Beran ◽  
Maria Lazo-Porras ◽  
Camille M. Mba ◽  
Jean Claude Mbanya
Keyword(s):  
Type 1 Diabetes ◽  
Diabetes Management ◽  
Multinational Companies ◽  
Global Perspective ◽  
Patient Access ◽  
Middle Income ◽  
Middle Income Countries ◽  
Wide Range ◽  
Pricing And Reimbursement

AbstractThe discovery of insulin in 1921 changed the prognosis for people with type 1 diabetes. A century later, availability and affordability of insulin remain a challenge in many parts of the globe. Using the WHO’s framework on understanding the life cycle of medicines, this review details the global and national challenges that affect patients’ abilities to access and afford insulin. Current research and development in diabetes has seen some innovations, but none of these have truly been game-changing. Currently, three multinational companies control over 95% of global insulin supply. The inclusion of insulin on the WHO’s Prequalification Programme is an opportunity to facilitate entry of new companies into the market. Many governments lack policies on the selection, procurement, supply, pricing and reimbursement of insulin. Moreover, mark-ups in the supply chain also affect the final price to the consumer. Whilst expenses related to diabetes are mostly covered by insurance in high-income countries, many patients from low- and middle-income countries have to pay out of their own pockets. The organisation of diabetes management within the healthcare system also affects patient access to insulin. The challenges affecting access to insulin are complex and require a wide range of solutions. Given that 2021 marks the centenary of the discovery of insulin, there is need for global advocacy to ensure that the benefits of insulin and innovations in diabetes care reach all individuals living with diabetes. Graphical abstract

scholarly journals Activation of the receptor tyrosine kinase RET improves long-term hematopoietic stem cell outgrowth and potency

Blood ◽  
2020 ◽  
Vol 136 (22) ◽  
pp. 2535-2547 ◽  
Author(s):  
W. Grey ◽  
R. Chauhan ◽  
M. Piganeau ◽  
H. Huerga Encabo ◽  
M. Garcia-Albornoz ◽  
...  
Keyword(s):  
Intracellular Signaling ◽  
Culture Conditions ◽  
Cellular Growth ◽  
Great Promise ◽  
Bone Marrow Niche ◽  
Hematopoietic Stem ◽  
Intracellular Signaling Pathway ◽  
Wide Range

Abstract Expansion of human hematopoietic stem cells (HSCs) is a rapidly advancing field showing great promise for clinical applications. Recent evidence has implicated the nervous system and glial family ligands (GFLs) as potential drivers of hematopoietic survival and self-renewal in the bone marrow niche; how to apply this process to HSC maintenance and expansion has yet to be explored. We show a role for the GFL receptor, RET, at the cell surface of HSCs in mediating sustained cellular growth, resistance to stress, and improved cell survival throughout in vitro expansion. HSCs treated with the key RET ligand/coreceptor complex, glial-derived neurotrophic factor and its coreceptor, exhibit improved progenitor function at primary transplantation and improved long-term HSC function at secondary transplantation. Finally, we show that RET drives a multifaceted intracellular signaling pathway, including key signaling intermediates protein kinase B, extracellular signal-regulated kinase 1/2, NF-κB, and p53, responsible for a wide range of cellular and genetic responses that improve cell growth and survival under culture conditions.

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