Proteinuria has been recognized as a common feature in many forms of chronic kidney disease (CKD). As traditional medications for proteinuric nephropathy, such as blockade of the renin-angiotensin system (RAS), has only achieved limited clinical success, more effective renoprotective strategies need to be explored. Bone marrow-derived mesenchymal stem/stromal cells (BM-MSCs) have recently shown promise as a therapeutic tool in acute kidney injury (AKI) models. The therapeutic potential of bone marrow-derived mesenchymal stem cells (BM-MSCs) in proteinuric nephropathy models is unknown. Using a co-culture model of human proximal tubular epithelial cells (PTECs) and BM-MSCs, I first examined the potential effect of BM-MSCs in albumin-induced pro-inflammatory response and epithelial-to-mesenchymal transition (EMT) in PTECs. The unstimulated BM-MSCs exerted moderate suppressive effect on tubular inflammation as only albumin-induced CCL-2 and CCL-5 expression was attenuated in PTECs. When concomitantly stimulated by albumin excess, however, BM-MSCs remarkably suppressed albumin-induced tubular IL-6, IL-8, TNF-α, CCL-2, and CCL-5 expression, suggesting albumin overloaded milieu to be a prerequisite for them to fully exhibit their anti-inflammatory effects. This effect was mediated via deactivation of tubular NF-κB signaling as BM-MSCs prevented the overexpression of p-IκB and nuclear translocation of NF-κB. In addition, albumin-induced tubular EMT, as shown by the loss of E-cadherin and induction of α-SMA, FN-1 and collagen IV in PTECs, was also prevented by BM-MSC co-culture. To dissect the mechanism of action, I next explored the paracrine factors secreted by BM-MSCs under an albumin-overloaded condition and studied their contribution to the protective effect on tubular inflammation and EMT. Albumin-overloaded BM-MSCs per se overexpressed 34 paracrine factors, of which hepatocyte growth factor (HGF) and TNFα-stimulating gene (TSG)-6 were regulated by P38 and NF-κB signaling. These paracrine factors suppressed both the proinflammatory and profibrotic phenotypes in albumin-induced PTECs. Neutralizing HGF and TSG-6 abolished the anti-inflammatory and anti-EMT effects of BM-MSC co-culture in albumin-induced PTECs, respectively. Finally, in albumin-overloaded mice, a well established murine model reminiscent of human CKD, treatment with mouse BM-MSCs markedly reduced BUN, tubular CCL-2 and CCL-5 expression, interstitial macrophage, α-SMA and collagen IV accumulation independent of changes in proteinuria, together with upregulated renal cortical expression of HGF. Exogenous BM-MSCs were detected in their kidneys by PKH-26 staining. Collectively, these in vitro and in vivo data suggest a modulatory effect of BM-MSCs on albumin-induced tubular inflammation and fibrosis and underscore a therapeutic potential of BM-MSCs for CKD in the future. / published_or_final_version / Medicine / Doctoral / Doctor of Philosophy

This item was digitized as part of the Million Books Project led by Carnegie Mellon University and supported by grants from the National Science Foundation (NSF). Cornell University coordinated the participation of land-grant and agricultural libraries in providing historical agricultural information for the digitization project; the University of Arizona Libraries, the College of Agriculture and Life Sciences, and the Office of Arid Lands Studies collaborated in the selection and provision of material for the digitization project.

This item was digitized as part of the Million Books Project led by Carnegie Mellon University and supported by grants from the National Science Foundation (NSF). Cornell University coordinated the participation of land-grant and agricultural libraries in providing historical agricultural information for the digitization project; the University of Arizona Libraries, the College of Agriculture and Life Sciences, and the Office of Arid Lands Studies collaborated in the selection and provision of material for the digitization project.

Cathepsin, matrix metalloproteinase (MMP) enzyme and tissue inhibitor of MMP (TIMP) distribution in J774 mouse macrophages has not been comprehensively studied. The distribution and vesicle regulation, trafficking and release of these is important, possibly suggesting drug targets for the therapeutic regulation of inflammatory disease and phagosomal killing of pathogenic organisms in J774 and other macrophages. Percentage immunofluorescence and ultrastructural enzyme and inhibitor distribution, together with LysoTracker (acidity) and lysosome-associated membrane proteins (LAMPs) colocalisation (both indicating late endosome or “lysosomal” association), western blot estimates of percentage processed- and unprocessed intracellular and secreted enzyme and inhibitor, and vesicle size was used to assign enzyme and inhibitor to “classical” vesicle types. Antibodies against TIMP-1 and TIMP-2 were raised and all antibodies characterised for this purpose. Together these were used to assign cathepsins H, S, D, B and L to possible secretory vesicles (±20 nm, non-acidic, LAMPs-negative, containing precursor enzymes) and identify at least 6 other endosome-“lysosome-like” vesicles. Cathepsin H appears to be present in classical early endosomes (±100 nm, non-acidic, LAMPs-negative) and cathepsin S in late endosomes(±50 nm, acidic, LAMPs-positive) and possibly “lysosomal” (“hybrid” or digestive organelles) (±150-200 nm, acidic, LAMPs-positive). Both cathepsins H and S, however, seem only reliable markers if used together with additional markers. Cathepsin D appearsmainly associated with “lysosomal” (“hybrid” or digestive organelles) (±150-200 nm, acidic, LAMPs-positive), possibly consisting of further subpopulations which requires furtherinvestigation e.g. labelling for LAMP-1 and LAMP-2 and cathepsin D. Cathepsins B and Lmay occur in late endosomes and/or hybrid organelles and “secretory lysosomes” containing cathepsins B, D and L may also exist (±30-50 nm, acidic, LAMPs-positive). The distribution of MMP-9, TIMP-1 and -2 in vesicles (non-acidic, LAMP-2-negative) thatappear novel and distinct from late endosome-“lysosome” vesicles were also demonstrated. In LPS-stimulated cells, the identity of the large (±450 nm), possible recycling endosomes (Rab11-positive, LAMPs-negative), containing colocalised MMP-9 and TIMP-1, needs investigation i.e., requires further verification with triple labelling and EM. Possible cell membrane and recycling endosome localisation of TIMP-2 needs confirmation with labelling of non-permeabilised cells and labelling for MT1-MMP and proMMP-2, respectively. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2007.

Purpose: To compare the safety and efficacy of intravitreal (IVT) and sub-Tenon (ST) triamcinolone acetonide for the treatment of refractory diffuse diabetic macular oedema. Method: 29 eyes of 22 patients with long-standing, diffuse diabetic macular oedema refractory to argon laser treatment were randomly assigned to a single 4mg injection of IVT triamcinolone acetonide or a 40mg sub-Tenon injection. Patients were subsequently monitored for six to nine months. Outcome measures were visual acuity, intraocular pressure, macular thickness on optical coherence tomography and adverse effects. Results: There was no significant improvement in visual acuity in either group. A transient decrease in macular thickness was found in the IVT group but not in the ST group. There were no significant adverse effects apart from a mild to moderate intra-ocular pressure rise found more frequently in the IVT group. Conclusion: IVT and ST triamcinolone acetonide injections for refractory diffuse diabetic macular oedema appear relatively safe and well-tolerated. IVT injection produces a significant temporary decrease in macular thickness in patients with long-standing diffuse diabetic macular oedema while ST injection does not. Neither intervention was shown to significantly improve visual acuity in this group of patients.

No description available.

Reduction of milk production following acute bovine mastitis causes important economic losses. In this study, two experiments were conducted to asses the ability of different antioxidants to prevent neutrophil (PMN)-induced mammary damage in acute bovine mastitis. First, a co-culture model composed of bovine mammary epithelial cell line (MAC-T cells) and bovine PMN activated by phorbol myristate acetate was used. Activated PMN release reactive oxygen species that are cytotoxic for bovine epithelial cells. Addition of dimethylthiourea or bathocuproinic acid did not induce any protective effect. On the other hand, addition of catechin, deferoxamine or glutathione ethyl ester (GEE) significantly reduced PMN-induced cytotoxicity in a dose-dependent manner as demonstrated by lower levels of released lactate dehydrogenase (LDH). The second experiment was undertaken with the last three antioxidants to evaluate their protective effects in vivo. A model of LPS-induced mastitis on dairy cows was used. The extent of cell damages was evaluated by measuring quarter milk levels of LDH and 4-methylumbelliferyl N-acetyl beta-D-glucosaminidase ( NAGase) at varying intervals before and after intramammary infusions of LPS, with or without antioxidants. Milk levels of haptoglobin and bovine serum albumin were also analysed. Catechin and GEE did not induce any protective effect whereas infusions of deferoxamine, a chelator of iron, decreased milk levels of LDH, NAGase and haptoglobin hence suggesting a protective effect against PMN-induced damage. Deferoxamine did not interfere with PMN migration into the mammary gland. Additionally, deferoxamine inhibited bacterial growth in vitro but did not affect PMN's ability to phagocytize live Escherichia coli. Overall, our results suggest that local infusion of deferoxamine may be an effective tool to protect mammary tissue against PMN-induced oxidative stress during bovine mastitis.

The purpose of this study was to determine duration time, breakage and apical displacement, whilst using six different endodontic filing systems to prepare molar teeth. A total of 96 molar teeth were used in the study, divided equally, ie 16 teeth per system selected randomly, totalling 48 canals per system. A standardised access cavity was prepared for all the teeth before selection. The canals were filed according to the manufacturers&rsquo / guidelines. The result showed that PROTAPER&reg / , K3&trade / and the combination of: HERO Shaper&reg / , HERO Apical&reg / and Endoflare&reg / (Referred from hereon as HERO System for convenience) were statistically faster than PROFILE&reg / and FlexMaster&reg / , which were in turn faster than AETTM. Although breakage did occur in K3&trade / and HERO System this was not deemed statistically significant. Apical displacement occurred in the form of Type 1 in the AETTM, PROFILE&reg / and HERO System, but once again this was not statistically significant. It was concluded that more aggressive cutting features such as a positive rake<br /> angle, pyramidal shaped tip, progressive taper and absence of radial lands, if present, could have enabled K3&trade / , HERO System and PROTAPER&reg / to have faster times, and in addition these features did not compromise these systems with regard to apical foramina transportation and breakage.

University of Macau / Institute of Chinese Medical Sciences

University of Macau / Institute of Chinese Medical Sciences