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  • Klintman, Daniel, et al. (författare)
  • Important role of p-selectin for leukocyte recruitment, hepatocellular injury, and apoptosis in endotoxemic mice.
  • 2004
  • Ingår i: Clinical and Diagnostic Laboratory Immunology. - : American Society for Microbiology. - 1071-412X. ; 11:1, s. 56-62
  • Tidskriftsartikel (refereegranskat)abstract
    • Leukocyte recruitment in the liver includes a two-step procedure in which selectin-dependent leukocyte rolling is a prerequisite for subsequent CD18-dependent leukocyte firm adhesion in postsinusoidal venules. However, the roles of the individual selectins in leukocyte rolling and adhesion, hepatocellular injury, and apoptosis remain elusive. Therefore, we examined the pathophysiological role of P-, E-, and L-selectin in male C57BL/6 mice challenged with lipopolysaccharide (LPS) and D-galactosamine (Gal) by use of intravital microscopy of the liver microcirculation. In control animals, administration of LPS-Gal provoked reproducible hepatic damage, including marked increases of leukocyte recruitment, liver enzymes, and hepatocyte apoptosis and reduced sinusoidal perfusion. Interestingly, pretreatment with an anti-P-selectin antibody (RB40.34) markedly reduced leukocyte rolling and firm adhesion by 65 and 71%, respectively. Moreover, interference with P-selectin function significantly improved sinusoidal perfusion and reduced the increase in liver enzymes by 49 to 84% in endotoxemic mice. Moreover, the activity of caspase-3 and the number of apoptotic hepatocytes were significantly reduced by 55 and 54%, respectively, in RB40.34-treated animals. In contrast, administration of an anti-E-selectin antibody (10E9.6) and an anti-L-selectin antibody (Mel-14) did not protect against endotoxin-induced leukocyte responses or hepatic injury. In conclusion, our novel findings document a principal role of P-selectin in mediating leukocyte rolling, a precondition to the subsequent firm adhesion of leukocytes in liver injury. Furthermore, our novel data demonstrate that inhibition of P-selectin function reduces hepatocellular injury and apoptosis, suggesting a causal relationship between leukocyte recruitment on one hand and hepatocellular injury and apoptosis on the other hand. Based on these findings, it is suggested that P-selectin may be an important therapeutic target in endotoxin-induced liver injury.
  • Klintman, Daniel, et al. (författare)
  • Leukocyte recruitment in hepatic injury: selectin-mediated leukocyte rolling is a prerequisite for CD18-dependent firm adhesion.
  • 2002
  • Ingår i: Journal of Hepatology. - : Elsevier. - 0168-8278. ; 36:1, s. 53-59
  • Tidskriftsartikel (refereegranskat)abstract
    • Background/Aims: This study was designed to examine the role of selectins and CD18 in leukocyte recruitment in hepatic injury induced by tumor necrosis factor-alpha (TNF-alpha) and galactosamine (Gal) in vivo.Methods: Intravital fluorescence microscopy of the hepatic microcirculation was used to quantify leukocyte-endothelium interactions provoked by 24 h of systemic TNF-alpha/Gal challenge in rats. Hepatic injury was evaluated with liver enzymes.Results: When administered after 24 h of TNF-alpha/Gal challenge, fucoidan, a selectin-function inhibitor, reduced leukocyte rolling by 69%, whereas firm adhesion was unaltered. In contrast, passive immunization against CD18 decreased leukocyte adhesion by 60%, whereas rolling remained unchanged. Notably, when administered prior to TNF-alpha/Gal, fucoidan attenuated both leukocyte rolling and adhesion, by 57 and 69%, respectively. Pretreatment with an anti-CD18 antibody decreased TNF-alpha/Gal-induced rolling and firm adhesion by 25 and 90%, respectively. Moreover, pretreatment with fucoidan and the anti-CD18 antibody both protected against TNF-alpha/Gal-induced increases in liver enzymes. For example, the pretreatments reduced alanine aminotransferase by 59 and 87%, respectively.Conclusions: Our data suggest that TNF-alpha/Gal-induced leukocyte rolling is selectin-mediated and a precondition for CD18-dependent firm adhesion in hepatic venules. Thus, reducing leukocyte recruitment by inhibition of selectins or CD18 may be useful to control TNF-alpha-induced liver injury.
  • Klintman, Daniel, et al. (författare)
  • Protective effect of Linomide on TNF-alpha-induced hepatic injury.
  • 2002
  • Ingår i: Journal of Hepatology. - : Elsevier. - 0168-8278. ; 36:2, s. 226-232
  • Tidskriftsartikel (refereegranskat)abstract
    • Background/Aims: Linomide is an immunomodulator that ameliorates several autoimmune and inflammatory diseases. We assessed the effect of Linomide on microvascular perfusion failure, leukocyte recruitment and hepatocellular injury induced by tumor necrosis factor alpha (TNF-alpha) and D-Galactosamine (Gal).Methods: After 3 days of Linomide pretreatment (1, 10 and 100mg/kg/day), rats were challenged with TNF-alpha/Gal for 24h. Microvascular perfusion, leukocyte--endothelium interactions in hepatic postsinusoidal venules and leukocyte sequestration in sinusoids were evaluated using intravital microscopy. Liver enzymes were measured spectrophotometrically.Results: Challenge with TNF-alpha/Gal significantly reduced sinusoidal perfusion, and increased leukocyte rolling, adhesion and liver enzymes. Interestingly, pretreatment with Linomide (10 and 100mg/kg/day) significantly reduced TNF-alpha/Gal-induced leukocyte rolling by 65 and 63%, and leukocyte adhesion by 87 and 84%, respectively. Moreover, Linomide (10 and 100mg/kg/day) decreased sinusoidal sequestration of leukocytes by 71 and 51%, and markedly improved sinusoidal perfusion. Moreover, Linomide reduced aspartate aminotransferase by 87--97%, and alanine aminotransferase by 79--96%. However, Linomide had no protective effect when administered concomitantly with TNF-alpha/Gal.Conclusions: These data demonstrate a dose-dependent inhibitory effect of Linomide on perfusion failure, leukocyte recruitment and hepatocellular injury provoked by TNF-alpha. Indeed, these findings suggest that Linomide may be an effective substance for protection of the liver in sepsis.
  • Koulaouzidis, Anastasios, et al. (författare)
  • Association Between Fecal Calprotectin Levels and Small-bowel Inflammation Score in Capsule Endoscopy : A Multicenter Retrospective Study
  • Ingår i: Digestive Diseases and Sciences. - : Springer. - 1573-2568. ; 61:7, s. 2033-2040
  • Tidskriftsartikel (refereegranskat)abstract
    • BACKGROUND: Accurate inflammation reporting in capsule endoscopy (CE) is important for diagnosis and monitoring of treatment of inflammatory bowel disease (IBD). Fecal calprotectin (FC) is a highly specific biomarker of gut inflammation. Lewis score (LS) was developed to standardize quantification of inflammation in small-bowel (SB) CE images.GOALS: Multicenter retrospective study aiming to investigate correlation between LS and FC in a large group of patients undergoing CE for suspected or known small-bowel IBD, and to develop a model for prediction of CE results (LS) based on FC levels.STUDY: Five academic centers and a district general hospital offering CE in UK, Finland, Sweden, Canada, and Israel. In total, 333 patients were recruited. They had small-bowel CE and FC done within 3 months.RESULTS: Overall, correlation between FC and LS was weak (r s: 0.232, P < 0.001). When two clinically significant FC thresholds (100 and 250 μg/g) were examined, the r s between FC and LS was 0.247 (weak) and 0.337 (moderate), respectively (P = 0.307). For clinically significant (LS ≥ 135) or negative (LS < 135) for SB inflammation, ROC curves gave an optimum cutoff point of FC 76 μg/g with sensitivity 0.59 and specificity 0.41.LIMITATIONS: Retrospective design.CONCLUSIONS: LS appears to show low correlation with FC as well as other serology markers of inflammation. FC does not appear to be a reliable biomarker for significant small-bowel inflammation. Nevertheless, FC level ≥ 76 μg/g may be associated with appreciable visual inflammation on small-bowel CE in patients with negative prior diagnostic workup.
  • Koulaouzidis, Anastasios, et al. (författare)
  • KID Project : an internet-based digital video atlas of capsule endoscopy for research purposes
  • Ingår i: Endoscopy International Open. - : Georg Thieme Verlag KG. - 2364-3722. ; 5:6, s. 477-483
  • Tidskriftsartikel (refereegranskat)abstract
    • BACKGROUND AND AIMS: Capsule endoscopy (CE) has revolutionized small-bowel (SB) investigation. Computational methods can enhance diagnostic yield (DY); however, incorporating machine learning algorithms (MLAs) into CE reading is difficult as large amounts of image annotations are required for training. Current databases lack graphic annotations of pathologies and cannot be used. A novel database, KID, aims to provide a reference for research and development of medical decision support systems (MDSS) for CE.METHODS: Open-source software was used for the KID database. Clinicians contribute anonymized, annotated CE images and videos. Graphic annotations are supported by an open-access annotation tool (Ratsnake). We detail an experiment based on the KID database, examining differences in SB lesion measurement between human readers and a MLA. The Jaccard Index (JI) was used to evaluate similarity between annotations by the MLA and human readers.RESULTS: The MLA performed best in measuring lymphangiectasias with a JI of 81 ± 6 %. The other lesion types were: angioectasias (JI 64 ± 11 %), aphthae (JI 64 ± 8 %), chylous cysts (JI 70 ± 14 %), polypoid lesions (JI 75 ± 21 %), and ulcers (JI 56 ± 9 %).CONCLUSION: MLA can perform as well as human readers in the measurement of SB angioectasias in white light (WL). Automated lesion measurement is therefore feasible. KID is currently the only open-source CE database developed specifically to aid development of MDSS. Our experiment demonstrates this potential.
  • Koulaouzidis, Anastasios, et al. (författare)
  • Novel experimental and software methods for image reconstruction and localization in capsule endoscopy
  • Ingår i: Endoscopy International Open. - : Georg Thieme Verlag KG. - 2364-3722. ; 6:2, s. 205-210
  • Tidskriftsartikel (refereegranskat)abstract
    • Background and study aims : Capsule endoscopy (CE) is invaluable for minimally invasive endoscopy of the gastrointestinal tract; however, several technological limitations remain including lack of reliable lesion localization. We present an approach to 3D reconstruction and localization using visual information from 2D CE images.Patients and methods : Colored thumbtacks were secured in rows to the internal wall of a LifeLike bowel model. A PillCam SB3 was calibrated and navigated linearly through the lumen by a high-precision robotic arm. The motion estimation algorithm used data (light falling on the object, fraction of reflected light and surface geometry) from 2D CE images in the video sequence to achieve 3D reconstruction of the bowel model at various frames. The ORB-SLAM technique was used for 3D reconstruction and CE localization within the reconstructed model. This algorithm compared pairs of points between images for reconstruction and localization.Results: As the capsule moved through the model bowel 42 to 66 video frames were obtained per pass. Mean absolute error in the estimated distance travelled by the CE was 4.1 ± 3.9 cm. Our algorithm was able to reconstruct the cylindrical shape of the model bowel with details of the attached thumbtacks. ORB-SLAM successfully reconstructed the bowel wall from simultaneous frames of the CE video. The "track" in the reconstruction corresponded well with the linear forwards-backwards movement of the capsule through the model lumen.Conclusion: The reconstruction methods, detailed above, were able to achieve good quality reconstruction of the bowel model and localization of the capsule trajectory using information from the CE video and images alone.
  • Laschke, MW, et al. (författare)
  • New experimental approach to study host tissue response to surgical mesh materials in vivo
  • 2005
  • Ingår i: Journal of Biomedical Materials Research. Part A. - : John Wiley and Sons Inc.. - 1552-4965. ; 74A:4, s. 696-704
  • Tidskriftsartikel (refereegranskat)abstract
    • Implantation of surgical meshes is a common procedure to increase abdominal wall stability in hernia repair. To improve biocompatibility of the implants, sophisticated in vivo animal models are needed to study inflammation and incorporation of biomaterials. Herein, we have established a new model that allows for the quantitative analysis of host tissue response and vascular ingrowth into surgical mesh materials in vivo. Ultrapro meshes were implanted into dorsal skinfold chambers of Syrian golden hamsters. Angiogenesis, microhemodynamics, microvascular permeability, and leukocyte-endothelial cell interaction of the host tissue were analyzed in response to material implantation over a 2-week period using intravital fluorescence microscopy. Mesh implantation resulted in a short-term activation of leukocytes, reflected by leukocyte accumulation and adherence in postcapillary venules. This cellular inflammatory response was accompanied by an increase of mac-romolecular leakage, indicating loss of integrity of venular endothelial cells. Angiogenesis started at day 3 after implantation by protrusion of capillary sprouts, originating from the host microvasculature. Until day 10, these sprouts interconnected with each other to form a new microvascular network. At day 14, the inflammatory response had disappeared and the vascular ingrowth was completed. Histology confirmed the formation of granulation tissue with adequate incorporation of the mesh filaments within the host tissue. We conclude that this novel model of surgical mesh implantation is a useful experimental approach to analyze host tissue response and vascular ingrowth of newly devised materials for hernia repair.
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