hm2095  (Zhongshan Golden Bridge Company)

Journal:

Article Title: Pharmacological targeting of C5a receptors during organ preservation improves kidney graft survival

doi: 10.1111/j.1365-2249.2008.03678.x

Figure Lengend Snippet: Impact of pharmacological complement 5a receptor (C5aR) targeting on kidney graft survival. Percentage survival rate of mice in each group as a function of post-transplantation time. Group A: 30 min cold ischaemia (CI) in University of Wisconsin (UW) solution (n = 25); group B: 2 h in UW solution without C5aRA (n = 21); group C: 2 h in UW solution with the addition of C5aRA (10−6 M) (n = 21). Pharmacological C5aR targeting improves graft survival rates significantly after 2 h of CI (P = 0·038; log-rank test).

Article Snippet: Complement 5a receptor immunohistochemistry on human biopsy material Paraffin-embedded sections were deparaffinized and rehydrated through two changes of xylene and graded alcohols, fixed with fresh 4% formaldehyde in PBS for 30 min at 4°C, blocked with normal goat serum and incubated with anti-human C5aR mAb (clone W17/1; Hycult Biotechnology) for 1 h at room temperature.

Techniques: Transplantation Assay

Journal:

Article Title: Pharmacological targeting of C5a receptors during organ preservation improves kidney graft survival

doi: 10.1111/j.1365-2249.2008.03678.x

Figure Lengend Snippet: Impact of complement 5a receptor (C5aR) targeting on ischaemia reperfusion injury-induced kidney damage. (a–e) Haematoxylin and eosin staining in transplanted (left panels) and non-transplanted kidneys (right panels). (a) 30 min cold ischaemia (CI), (b) 2 h CI without C5aRA and (c) 2 h CI in the presence of C5aRA. Small arrows depict glomeruli and large arrows indicate regions of haemorrhage. Figures are reduced from an original magnification of 20×. (d) Quantification of tissue damage in transplanted (left panel) and non-transplanted (right panel) kidneys. A tissue damage score was determined on a scale of 0–3 (none, mild, moderate and severe) as outlined in Materials and methods, with a maximum possible score of 9 = severest damage being attainable. (e) Spatial distribution of damage in transplanted (left panel) and non-transplanted kidneys (right panel). (f) Relative kidney damage shown as a composite damage score calculating the product of each individual damage score with its corresponding area of damage. All values were determined 72 h post-transplantation (n = 6–12).

Article Snippet: Complement 5a receptor immunohistochemistry on human biopsy material Paraffin-embedded sections were deparaffinized and rehydrated through two changes of xylene and graded alcohols, fixed with fresh 4% formaldehyde in PBS for 30 min at 4°C, blocked with normal goat serum and incubated with anti-human C5aR mAb (clone W17/1; Hycult Biotechnology) for 1 h at room temperature.

Techniques: Staining, Transplantation Assay

Journal:

Article Title: Pharmacological targeting of C5a receptors during organ preservation improves kidney graft survival

doi: 10.1111/j.1365-2249.2008.03678.x

Figure Lengend Snippet: Impact of complement 5a receptor (C5aR) targeting on ischaemia reperfusion injury-induced tubular apoptosis. (a–c) Transferase-mediated dUTP nick-end labelling (TUNEL)-positive tubular epithelial cells in transplanted (left panels) and non-transplanted kidneys (right panels). (a) 30 min cold ischaemia (CI). (b) 2 h CI without C5aRA. (c) 2 h CI in the presence of C5aRA. TUNEL-positive cells in the transplanted kidneys of groups a–c animals are depicted by arrows (left panels). Figures are reduced from an original magnification of 20×. (c, d) Quantification of apoptosis in tubular epithelial cells of transplanted (c) and non-transplanted (d) kidneys (number of TUNEL-positive cells/100 tubular cells in each of five high-power fields). Groups were compared by one way analysis of variance to determine statistical differences between treatment groups (see Material and methods). All values were determined 72 h post transplantation (n = 6–12).

Article Snippet: Complement 5a receptor immunohistochemistry on human biopsy material Paraffin-embedded sections were deparaffinized and rehydrated through two changes of xylene and graded alcohols, fixed with fresh 4% formaldehyde in PBS for 30 min at 4°C, blocked with normal goat serum and incubated with anti-human C5aR mAb (clone W17/1; Hycult Biotechnology) for 1 h at room temperature.

Techniques: TUNEL Assay, Transplantation Assay

Journal:

Article Title: Pharmacological targeting of C5a receptors during organ preservation improves kidney graft survival

doi: 10.1111/j.1365-2249.2008.03678.x

Figure Lengend Snippet: Impact of complement 5a receptor (C5aR) targeting on ischaemia reperfusion injury-induced C5aR expression and tissue inflammation. (a–c) C5aR expression in transplanted (left panels) and non-transplanted kidneys (right panels). (a) 30 min cold ischaemia (CI). Black arrows indicate C5aR positive macrophages; white arrows indicate C5aR positive tubular epithelial cells. (b) 2 h CI without C5aRA. (c) 2 h CI in the presence of C5aRA. Figures are reduced from an original magnification of 20×. (d) C5aR; (e) tumour necrosis factor-α; and (f) macrophage inflammatory protein-2/CXCL2 mRNA expression levels, quantified by real-time polymerase chain reaction in transplanted (left panel) and non-transplanted (right panel) kidneys respectively. The mRNA expression between the indicated treatment groups was compared. All values were determined 72 h post-transplantation (n = 6–12).

Article Snippet: Complement 5a receptor immunohistochemistry on human biopsy material Paraffin-embedded sections were deparaffinized and rehydrated through two changes of xylene and graded alcohols, fixed with fresh 4% formaldehyde in PBS for 30 min at 4°C, blocked with normal goat serum and incubated with anti-human C5aR mAb (clone W17/1; Hycult Biotechnology) for 1 h at room temperature.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Transplantation Assay

Journal:

Article Title: Pharmacological targeting of C5a receptors during organ preservation improves kidney graft survival

doi: 10.1111/j.1365-2249.2008.03678.x

Figure Lengend Snippet: Complement 5a receptor (C5aR) expression in cadaveric donor (CAD) and living-related donor (LRD) and its correlation with cold ischaemia time, kidney function and tubular cell apoptosis. (a, b) C5aR expression in glomeruli, tubules and the interstitium of human allografts from LRD (a, upper panels) or CAD (b, lower panels). C5aR staining is depicted by the white arrowheads in each panel. (c) Quantitative evaluation of C5aR expression in grafts from CAD (n = 13) and LRD (n = 12). Groups were compared by analysis of variance to determine statistical differences between treatment groups (see Material and methods). (d) Correlation between cold ischaemia time (CIT) and peak serum creatinine (sCr) levels (r = 0·97, P < 0·001). (e–g) Correlations between C5aR staining intensity (number of C5aR positive tubules/field) and peak sCr levels (r = 0·96, P < 0·001) (e), duration of CIT (r = 0·82, P < 0·001) (f) and the frequency of apoptotic cells in the allograft (r = 0·78, P = 0·001) (g).

Article Snippet: Complement 5a receptor immunohistochemistry on human biopsy material Paraffin-embedded sections were deparaffinized and rehydrated through two changes of xylene and graded alcohols, fixed with fresh 4% formaldehyde in PBS for 30 min at 4°C, blocked with normal goat serum and incubated with anti-human C5aR mAb (clone W17/1; Hycult Biotechnology) for 1 h at room temperature.

Techniques: Expressing, Staining