Early degradation of type IX and type II collagen with the onset of experimental inflammatory arthritis

OBJECTIVE: To determine whether following the onset of intraarticular inflammation, there is early damage to articular cartilage, specifically to types II and IX collagen, and the proteoglycan (PG) aggrecan, and whether measurement of the degradation products of these molecules in synovial fluid (SF) and serum may permit the detection of cartilage damage. METHODS: A rabbit model of rheumatoid arthritis, antigen (ovalbumin)-induced arthritis, was studied. Articular cartilage samples were analyzed by immunoassays for total type II collagen content, its denaturation and cleavage by collagenases, and for type IX collagen content. PG content was determined by colorimetric assay. In serum and SF, total PG content and collagenase-generated peptides of type II collagen were measured. RESULTS: After 6 days, both the PG content and the NC4 domain of type IX collagen were reduced in femoral and tibial cartilage, concomitant with the onset of arthritis. In only the tibial cartilage did this reduction in PG persist up to day 20. However, denatured type II collagen was increased in all cartilage samples, but only on day 20. In SF, the PG content was significantly reduced on day 20, and products of type II collagen cleavage by collagenase were significantly increased on both day 6 and day 20. CONCLUSION: This study, which is the first of its kind examining changes in both types II and IX collagen and PG content, reveals early damage to both types of collagen as well as to PG in articular cartilage samples following induction of joint inflammation. SF analyses reveal this early damage and may be of value in the study and treatment of inflammatory arthritic diseases such as rheumatoid arthritis.     

Animal models for studying the etiology and treatment of low back pain

Chronic low back pain is a major cause of disability and health care costs. Effective treatments are inadequate for many patients. Animal models are essential to further understanding of the pain mechanism and testing potential therapies. Currently, a number of preclinical models have been developed attempting to mimic aspects of clinical conditions that contribute to low back pain (LBP). This review focused on describing these animal models and the main behavioral tests for assessing pain in each model. Animal models of LBP can be divided into the following five categories: Discogenic LBP, radicular back pain, facet joint osteoarthritis back pain, muscle‐induced LBP, and spontaneous occurring LBP models. These models are important not only for enhancing our knowledge of how LBP is generated, but also for the development of novel therapeutic regimens to treat LBP in patients. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1305–1312, 2018.

     

Gender differences in relation to sputum submission and smear-positive pulmonary tuberculosis in Malawi.

OBJECTIVE: To examine gender differences in sputum submission and sputum smear positivity. METHODS: Laboratory registers in all diagnostic units in eight districts in Malawi were examined for the years 1995 and 1996. RESULTS: During a 12-month period (averaged between 1995 and 1996), 26,624 new TB suspects submitted sputum samples, 3282 of which (12.3%) were smear-positive. Significantly more males submitted sputum (52%) compared with females (48%), and significantly more males (53%) were smear-positive compared with females (47%, P < 0.05). Rates of sputum submission per 100,000 adults were also significantly higher for males (1203) than females (1032). CONCLUSION: In Malawi, fewer females are submitting sputum samples and are being diagnosed with smear-positive TB compared with males.     

Limitations of using aggrecan and type X collagen as markers of chondrogenesis in mesenchymal stem cell differentiation.

The study was initially designed to differentiate human bone marrow-derived mesenchymal stem cells (MSC) into chondrocyte-like cells, for use in tissue engineering. We cultured MSCs in defined chondrogenic medium as pellet cultures supplemented with transforming growth factor (TGF)-beta1 or -beta3 and dexamethazone, as they are commonly used to promote in vitro chondrogenesis. Markers of chondrogenesis used were type II collagen and aggrecan, with type X collagen being used as a marker of late-stage chondrocyte hypertrophy (associated with endochondral ossification). Our results show that aggrecan is constitutively expressed by MSCs and that type X collagen is expressed as an early event. Furthermore, we found that type X collagen was expressed before type II collagen in some cases. This is surprising because it is understood that stem cells have to be differentiated into chondrocytes before they can become hypertrophic. Thus, caution must be exercised when using aggrecan and type X collagen as markers for chondrogenesis and chondrocyte hypertrophy, respectively, in association with stem cell differentiation from this source.     

Changes in Escherichia coli resistance to co-trimoxazole in tuberculosis patients and in relation to co-trimoxazole prophylaxis in Thyolo,Malawi

In Thyolo district, Malawi, an operational research study is being conducted on the efficacy and feasibility of co-trimoxazole prophylaxis in preventing deaths in HIV-positive patients with tuberculosis (TB). A series of cross-sectional studies were carried out to determine i) whether faecal Escherichia coli (E.coli) resistance to co-trimoxazole in TB patients changed with time and ii) whether the resistance pattern was different in HIV positive TB patients who were taking co-trimoxazole prophylaxis. Co-trimoxazole resistance among E.coli isolates in TB patients at the time of registration was 60% in 1999 and 77% in 2001 (p<0.01). Resistance was 89% among HIV-infected TB patients (receiving co-trimoxazole), while in HIV negative patients (receiving anti-TB therapy alone) it was 62% (p<0.001). The study shows a significant increase of E.coli resistance to co-trimoxazole in TB patients which is particularly prominent in HIV infected patients on co-trimoxazole prophylaxis. Since a high degree of plasmid-mediated transfer of resistance exists between E.coli and the Salmonella species, these findings could herald limitations on the short and long term benefits to be anticipated from the use of co-trimoxazole prophylaxis in preventing non-typhoidal salmonella bacteraemia and enteritis in HIV infected TB patients in Malawi.     

Group reorientation and migration of amphiphilic polymer bearing phosphorylcholine functionalities on surface of cellular membrane mimicking coating

Amphiphilic polymers bearing phosphorylcholine (PC) groups can form films of interfacial structure similar to that of the outer membrane of living cells. The films, as prepared, present PC groups to the external aqueous environment and exhibit good biocompatibility. However, under certain conditions, the surface structure can change irreversibly due to the reorientation and deep migration of the surface groups. X-ray photoelectron spectroscopy (XPS), dynamic contact angle measurements, and cell culture experiments were used to investigate the reorientation and migration of the surface groups of an amphiphilic PC-polymer coating. When the polymer surface is immersed into or drawn out of water, significant reorientation and group migration occurs, as suggested by the large difference between the advancing and receding contact angles. Angle-resolved XPS measurements indicate that the hydrophobic groups move to the air/film interface while the hydrophilic groups migrate towards the bulk of the polymer coating. Long periods of aging may result in irreversible changes of the surface structure and decrease the biocompatibility of the materials.     

Effect of cobalt and chromium ions on bcl-2, bax, caspase-3, and caspase-8 expression in human U937 macrophages

The bcl-2 and caspase families of proteins play a central role in the modulation of apoptosis. The purpose of this study was to analyze the effect of Co(2+) and Cr(3+) ions on the expression of bcl-2, bax, caspase-3 and caspase-8 to better understand the mechanisms leading to ion-induced apoptosis in macrophages. U937 human macrophages were exposed to Co(2+) and Cr(3+) ions. The expression of proteins was measured by Western blot while caspase activities were measured by colorimetric assay. Results show that Co(2+) ions inhibited bcl-2 expression with significant effect (p<0.05) after 16 h and a maximal 52% inhibitory effect after 24 h. Co(2+) stimulated bax expression with a significant stimulation (p<0.05) after 8 h and a maximal 1.75-fold increase after 16 h. Co(2+) also stimulated the expression of the active fragment of caspase-3 as well as caspase-3 activity maximal increase after 24 h. Co(2+) ions had no effect on caspase-8 expression or activity.Cr(3+) ions inhibited bcl-2 expression with significant effect (p<0.05) after 16 h and a maximal 43% inhibitory effect after 24 h. Cr(3+) stimulated bax expression with significant stimulation (p<0.01) after 8h and a maximal 2.25-fold increase after 24 h. Cr(3+) ions also stimulated the expression of the active fragments of caspase-3 and -8, as well as the activities of both proteases. The effect of Cr(3+) ions on the expression of both caspase active fragments was maximal after 16 h incubation. In conclusion, our results suggest that the modulation of the expression of proteins from the bcl-2 and the caspase families of proteins are implicated in the induction of macrophage apoptosis by Co(2+) and Cr(3+) ions.     

Promotion of U937 cell adhesion on polypropylene surfaces bearing phosphorylcholine functionalities

Phosphorylcholine (PC) groups were grafted onto ammonia plasma-treated biaxially-oriented polypropylene (BOPP) surfaces, via (a) reductive amination of phosphorylcholine glyceraldehyde and (b) a two-step procedure involving the chemical amplification of surface amine groups with tris(2-aminoethyl amine) and subsequent reductive amination of phosphorylcholine glyceraldehyde. The occurrence of grafting was ascertained by X-ray photoelectron spectroscopy (XPS) and attenuated total reflectance Fourier-transform infrared (ATR-FT-IR) spectroscopy. The wettability of PC-modified surfaces was assessed by dynamic contact-angle measurements using the Wilhelmy plate method. Human U937 macrophages adhered and proliferated to a significantly larger extent on PC-modified surfaces, compared to unmodified or ammonia plasma-modified BOPP.