Bacterial lipopolysaccharides form procollagen-endotoxin complexes that trigger cartilage inflammation and degeneration: implications for the development of rheumatoid arthritis

Introduction

We have previously reported that bacterial toxins, especially endotoxins such as lipopolysaccharides (LPS), might be important causative agents in the pathogenesis of rheumatoid arthritis (RA) in an in vitro model that simulates the potential effects of residing in damp buildings. Since numerous inflammatory processes are linked with the nuclear factor-κB (NF-κB), we investigated in detail the effects of LPS on the NF-κB pathway and the postulated formation of procollagen-endotoxin complexes.

Methods

An in vitro model of human chondrocytes was used to investigate LPS-mediated inflammatory signaling.

Results

Immunoelectron microscopy revealed that LPS physically interact with collagen type II in the extracellular matrix (ECM) and anti-collagen type II significantly reduced this interaction. BMS-345541 (a specific inhibitor of IκB kinase (IKK)) or wortmannin (a specific inhibitor of phosphatidylinositol 3-kinase (PI-3K)) inhibited the LPS-induced degradation of the ECM and apoptosis in chondrocytes. This effect was completely inhibited by combining BMS-345541 and wortmannin. Furthermore, BMS-345541 and/or wortmannin suppressed the LPS-induced upregulation of catabolic enzymes that mediate ECM degradation (matrix metalloproteinases-9, -13), cyclooxygenase-2 and apoptosis (activated caspase-3). These proteins are regulated by NF-κB, suggesting that the NF-κB and PI-3K pathways are involved in LPS-induced cartilage degradation. The induction of NF-κB correlated with activation of IκBα kinase, IκBα phosphorylation, IκBα degradation, p65 phosphorylation and p65 nuclear translocation. Further upstream, LPS induced the expression of Toll-like receptor 4 (TLR4) and bound with TLR4, indicating that LPS acts through TLR4.

Conclusion

These results suggest that molecular associations between LPS/TLR4/collagen type II in chondrocytes upregulate the NF-κB and PI-3K signaling pathways and activate proinflammatory activity.     

The K-Segments of the Wheat Dehydrin DHN-5 are Essential for the Protection of Lactate Dehydrogenase and β-Glucosidase Activities In Vitro

The wheat dehydrin DHN-5 has been previously shown to exhibit heat protecting effect on enzymatic activities. In order to understand the molecular mechanism by which DHN-5 exerts its protective function, we performed an approach to dissect the functional domains of DHN-5 responsible for this feature. In two distinct enzymatic assays, we found that the truncated forms of DHN-5 containing only one K- or two K-segments are able to protect albeit to less extent than the wild type protein, lactate dehydrogenase and β-glucosidase against damage induced by various stresses in vitro. However, the YS- and Φ-segments alone have no protective effects on these enzymes. Therefore, our study provides the evidence that the protective function of DHN-5 seems to be directly linked to its K-segments which through their amphipatic α-helical structure, may act to prevent protein aggregation.     

A Dermal Gel Made of Rutilus Kutum Skin Collagen-Chitosan for Deep Burn Healing

International Journal of Peptide Research and Therapeutics - Natural compounds extracted from marine organisms consisting of biological active materials like collagen provide a major source of...     

Phylogenetic Analysis of Clinically Relevant Fusarium Species in Iran

Mycopathologia - Fungi of the genus Fusarium are well known as major plant pathogens but also cause a broad spectrum of human infections. Sixty-three clinical isolates, collected during...     

Characterization of Aqueous Dispersions and Gels Made of Sodium Caseinate and Basil Seed Gum: Phase Behavior,Rheology, and Microstructure

The interactions between sodium caseinate (NaCas) and basil seed gum (BSG) in the presence of calcium chloride (CaCl₂) were investigated. The phase behavior of the mixed aqueous dispersions and their gels revealed a homogeneous mixture, obtained at the higher concentrations of both CaCl₂ and BSG. The Herschel-Bulkley model sufficiently fitted the flow behavior of the mixture solution data. Apparent viscosity increased significantly (p < 0.05) by increasing the concentration of BSG, where the addition of CaCl₂ had no significant effect on the viscosity of the samples (p > 0.05). Furthermore, there was an increase in thixotropy due to the higher concentrations of BSG and CaCl₂. Based on the frequency sweep test, at the low frequencies, a more gel-like behavior was observed in the case of the higher concentrations of either BSG or CaCl₂. The rheological and SEM data suggested that the stronger structure of NaCas-BSG gel in the presence of the higher concentrations of CaCl₂ was related to the induction of complex formation between the two biopolymers.

     

Countrywide Survey for MERS-Coronavirus Antibodies in Dromedaries and Humans in Pakistan

Middle East Respiratory Syndrome Coronavirus (MERS-CoV) is a zoonotic pathogen capable of causing severe respiratory disease in humans. Although dromedary camels are considered as a major reservoir host, the MERS-CoV infection dynamics in camels are not fully understood. Through surveillance in Pakistan, nasal (n = 776) and serum (n = 1050)samples were collected from camels between November 2015 and February 2018. Samples were collected from animal markets, free-roaming herds and abattoirs. An in-house ELISA was developed to detect IgG against MERS-CoV. A total of 794 camels were found seropositive for MERS-CoV. Prevalence increased with the age and the highest seroprevalence was recorded in camels aged [ 10 years (81.37%) followed by those aged 3.1–10 years (78.65%) and B 3 years (58.19%).Higher prevalence was observed in female (78.13%) as compared to male (70.70%). Of the camel nasal swabs, 22 were found to be positive by RT-qPCR though with high Ct values. Moreover, 2,409 human serum samples were also collected from four provinces of Pakistan during 2016–2017. Among the sampled population, 840 humans were camel herders.Although we found a high rate of MERS-CoV antibody positive dromedaries (75.62%) in Pakistan, no neutralizing antibodies were detected in humans with and without contact to camels.     

Approximation of ground water quality for microbial and chemical contamination

Present study was designed to obtain estimation about ground water quality of Bhimber, Azad Jammu and Kashmir (AJK), Pakistan. A total of 12 water samples were collected from different localities of study area to analyze for various physicochemical and biological parameters i.e. namely temperature, pH, turbidity, color, odor, taste, electric conductivity (EC), total dissolved solids (TDS), total hardness (Calcium + Magnesium), chloride, arsenic, phosphate, lead, ammonium ion, nitrite, Fecal coliform and Escherichia coli. Results exposed that all ground water samples of study area were grossly contaminated with pathogenic microorganisms like E. coli and Fecal coliform except one water sample that was obtained from community filter plant Samahni Chowk site. Besides it, values of some physicochemical water quality determining parameters also deviated from recommended limits of World Health Organization (WHO). Chloride ion concentration was little below the prescribed limits in almost all water samples. It has been proven that consumption of un-safe drinking water is one of the major cause of prevalence of water borne diseases like diarrhea, gastroenteritis, typhoid fever and malaria etc. in study area. Community water supply and sanitation projects should be encouraged; government should provide filter plants in all regions of the country so that people could have easy approach to safe drinking water.