Atrial natriuretic peptide preconditioning protects against hepatic preservation injury by attenuating necrotic and apoptotic cell death

Size: px
Start display at page:

Download "Atrial natriuretic peptide preconditioning protects against hepatic preservation injury by attenuating necrotic and apoptotic cell death"

Transcription

1 Journal of Hepatology 39 (2003) Atrial natriuretic peptide preconditioning protects against hepatic preservation injury by attenuating necrotic and apoptotic cell death Tobias Gerwig 1,2, Herbert Meibner 3, Manfred Bilzer 2, Alexandra K. Kiemer 1, Hans Arnholdt 3, Angelika M. Vollmar 1, Alexander L. Gerbes 2, * 1 Department of Pharmacy, Center of Drug Research, University of Munich, Klinikum Grosshadern, Marchioninistrasse 15, D Munich, Germany 2 Department of Medicine II, University of Munich, Klinikum Grosshadern, Marchioninistrasse 15, D Munich, Germany 3 Institute of Pathology, University of Munich, Klinikum Grosshadern, Marchioninistrasse 15, D Munich, Germany See Editorial, pages Background/Aims: Preconditioning of livers with the atrial natriuretic peptide (ANP) markedly reduces hepatic ischemia-reperfusion injury. Aim of this study was to characterize the influence of ANP preconditioning on necrotic and apoptotic cell death and on proliferation. Methods: Rat livers were perfused with Krebs-Henseleit buffer with or without ANP or its second messenger analogue 8-Bromo cyclic guanosine monophosphate (8-Br cgmp) for 20 min, stored in cold University of Wisconsin solution (24 h), and reperfused for up to 120 min. Apoptosis and necrosis were determined using biochemical and morphological criteria, proliferation was assessed by Ki67 histochemistry. Results: Apoptosis peaked after 24 h of cold ischemia. Preconditioning with both ANP and 8-Br-cGMP significantly reduced caspase-3-like activity and the number of triphosphate nick-end labelling-positive cells. Reduction of apoptosis was significant for hepatocytes, but not for endothelial cells. After ischemia, degenerative cell changes were clearly reduced in ANP pretreated livers. After reperfusion, ANP preconditioning led to a significant reduction of necrotic hepatocytes and endothelial cells in periportal zones. Cell proliferation was not affected by preconditioning. Conclusions: ANP reduces necrotic and apoptotic cell death without affecting the proliferation status. The protection takes place mainly in the periportal area and seems to be most prominent against necrosis of hepatocytes and endothelial cells during reperfusion. q 2003 European Association for the Study of the Liver. Published by Elsevier Science B.V. All rights reserved. Keywords: Hormonal preconditioning; Liver transplantation; cgmp; Ischemia-reperfusion 1. Introduction Ischemia-reperfusion injury (IRI) is the main cause of primary dysfunction or non-function after liver resection or transplantation. Therefore, it is of great clinical interest to elucidate the underlying pathomechanisms and to develop protective strategies [1]. IRI is caused by a combination of cellular changes in the ischemic period and the consecutive reperfusion period Received 28 August 2002; received in revised form 18 March 2003; accepted 5 May 2003 * Corresponding author. Tel.: þ ; fax: þ address: gerbes@med2.med.uni-muenchen.de (A.L. Gerbes). where blood flow and oxygen supply is restored. In the ischemic phase, reduced energy status leads to a breakdown of energy-dependent metabolic pathways and transport processes, resulting in perturbation of ion homeostasis and activation of proteases [2 5]. In organ transplantation, hypothermic storage itself causes negative effects like cell swelling and calcium alterations despite the beneficial properties of low temperatures to prolong the possible storing time by reducing the metabolic rate [6,7]. Sinusoidal endothelial cells (SEC) seem to be more susceptible to cold ischemia than hepatocytes [8] and contribute to reduced organ viability by detaching from their cellular matrix at the beginning of reperfusion [9]. When oxygen supply is restored in the reperfusion /03/$30.00 q 2003 European Association for the Study of the Liver. Published by Elsevier Science B.V. All rights reserved. doi: /s (03)00240-x

2 342 T. Gerwig et al. / Journal of Hepatology 39 (2003) period, Kupffer cells are rapidly activated and are regarded as central mediators of hepatic IRI [1,10,11]. They release reactive oxygen species and activate a variety of mediators including platelet activating factor, interleukins, tumor necrosis factor (TNF)-a, transcription factors, and adhesion molecules [12 15], leading to microcirculatory failure and manifestation of IRI. IRI was originally considered to be due to a necrotic type of cell death. However, there is increasing evidence for a role of apoptotic cell death in IRI of heart and brain [16,17]. Apoptosis is defined by morphological criteria [18]. These comprise nuclear fragmentation with chromatin condensation and cell shrinkage followed by fragmentation into membrane bound apoptotic bodies. Recent studies reported apoptotic cell death of SEC and hepatocytes during reperfusion of the liver [19,20]. The significance of apoptosis for warm or cold [21,22] hepatic IRI, however, has been questioned. Recently, we demonstrated that hormonal preconditioning with the atrial natriuretic peptide (ANP) protects the liver from IRI after both warm and cold [23,24] ischemia. ANP mediates its hepatoprotective action via the guanylatecyclase coupled A-receptor, attenuating the inflammatory response of IRI [14]. The activation of a heat shock response might contribute to its anti-inflammatory potential [25]. Stimulated by the increasing discussion of the role of apoptosis for hepatic IRI we here focussed on the occurrence of apoptotic compared to necrotic cell death using the model of the isolated perfused rat liver. We characterized apoptotic cell death in addition to necrotic cell death in hepatocytes and SEC and determined the effects of ANP. In addition, we assessed the hepatocyte proliferation state to investigate a potential effect of preconditioning on liver regeneration. 2. Materials and methods 2.1. Materials Rat ANP was obtained from Calbiochem/Novabiochem (Bad Soden, Germany), Narcoren w from Merial (Hallbergmoos, Germany), Complete w from Roche Diagnostics GmbH (Mannheim, Germany), N-acetyl-Asp-Glu-Val-Asp-7-amino-4-trifluoromethyl-coumarin (Ac-DEVD-AFC) from Biosource (Hamburg, Germany), anti-caspase-3 antibody (sc-7148, rabbit) from Santa Cruz Biotechnology (Heidelberg, Germany), and Pefabloc w SC from Biomol GmbH (Hamburg, Germany). All other materials were purchased from Sigma (Taufkirchen, Germany) Liver perfusion Male Sprague Dawley rats weighing g were purchased from Charles River GmbH (Sulzfeld, Germany) and housed in a climatized room with a 12-h light-dark cycle. The animals had free access to chow (Standard-Diet, Altromin 1314 Lage, Germany) and water. After anaesthetizing the animals with Narcoren w (50 mg/kg body weight, intraperitoneally), the livers were perfused via the portal vein in situ with a bicarbonate buffered Krebs-Henseleit (KH) solution (ph 7.4, 378C) gassed with 95% O 2 and 5% CO 2, as described previously [14,23,24]. The perfusion medium was pumped through the livers at a constant flow rate of ml min 21 g liver 21 in a non-recirculating fashion. After 30 min of perfusion with KH buffer, livers were perfused with cold (48C) University of Wisconsin (UW) solution (DuPont, Bad Homburg, Germany) for 1 min. The organs were then kept in UW solution at 48C for 24 h. Pretreatment was performed by adding ANP (200 nm) or 8-Br-cGMP (50 mm) to the pre-ischemic perfusate for 20 min until ischemia and to the storage solution. Livers were reperfused with KH buffer for up to 2 h. At the indicated times, livers were snap-frozen and stored at 2808C until further analysis. Unless stated otherwise, four to five independent experiments were performed. All animals received humane care. The study was registered with the local animal welfare committee Determination of apoptosis and necrosis To distinguish apoptotic from necrotic cell death, commonly used biochemical methods in combination with morphological evaluation were used [21,26]. As parameters for apoptotic cell death, caspase-3-like activity in combination with caspase-3 protein processing and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling (TUNEL) were determined. Morphological criteria were cell shrinkage, chromatin condensation and margination, and the occurrence of apoptotic bodies. Necrotic cell death was characterized morphologically by hematoxylin and eosin (HE) staining and trypan blue uptake into injured cells Measurement of caspase-3-like activity Cytosolic extracts from liver tissue were prepared by homogenization in hypotonic extraction buffer (25 mmol/l HEPES, ph 7.5; 5 mmol/l MgCl 2 ; 1 mmol/l EGTA; 1 mmol/l Pefablock w SC; and pepstatin, leupeptin, and aprotinin, 1 mg/ml each) in a Potter device (Braun, Melsungen, Germany.). Homogenates were subsequently centrifuged, and supernatants were stored at 2808C. The fluorometric Ac-DEVD-AFC cleavage assay was carried out on microtiter plates (Greiner, Nürtingen, Germany) according to the method originally described by Nicholson et al. [27]. Cytosolic extracts (10 ml,,1 mg/ml protein) were diluted 1:10 with substrate buffer (55 mmol/l of fluorogenic substrate N-acetyl-Asp-Glu-Val-Asp-7-amino- 4-trifluoromethyl-coumarin in 50 mmol/l HEPES, ph 7.4, 1% sucrose, 0.1% CHAPS, 10 mmol/l dithiothreitol). Blanks contained 10 ml of extraction buffer and 90 ml of substrate buffer. Generation of free 7-amino-4-trifluoro-methylcoumarin at 378C was kinetically determined using a fluorometer plate reader (excitation, 385 nm; emission, 505 nm) (Fluostar, BMG GmbH, Offenburg, Germany). Enzyme activity was calculated using an external AFC standard curve. The highest enzyme activity measured was set to 100%. Protein concentrations of the samples were estimated with the Pierce Assay (Pierce, Rockford, USA). Control experiments confirmed that the activity was linear with time and with protein concentration under the conditions described above Western blot analysis of caspase-3 processing Deep-frozen liver samples were lysed in 150 mm NaCl, 50 mm Tris HCl, ph 7.5, 1% Nonidet P-40, 0.25 deoxycholate, 0.1% sodium dodecyl sulfate (SDS) supplemented with a protease inhibitor cocktail (Complete w ). The protein concentration was determined as described by Lowry et al. [28]. UW solution in the dilution employed did not affect protein determination. Equal amounts of liver lysates (50 mg) were loaded onto an SDS/polyacrylamide gel electrophoresis (10%), electroblotted onto a PVDF membrane (Millipore, Eschwege, Germany), and caspase-3 precursor (CPP32) and proteolytic active subunit (p17) were detected using a specific rabbit polyclonal anti-caspase-3 antibody (sc-7148, rabbit) from Santa Cruz (Heidelberg, Germany) and the Renaissance A chemiluminescence reagent plus (NEN Lifescience, Cologne, Germany) with a KODAK Image Station 440CF (Eastman Kodak Company, Rocester, New York, USA) TUNEL analysis To determine strand brakes of genomic DNA cells were stained with the TUNEL-assay [29]: at indicated time points liver samples were taken and slices were immediately fixed in buffered formalin solution (4% formaldehyde). Samples were embedded in paraffin and cut into 6 mm sections. For evaluation of apoptotic cells liver sections were stained with

3 T. Gerwig et al. / Journal of Hepatology 39 (2003) Statistics All data are expressed as mean and standard error of mean. The significance of differences between experimental groups was determined by one-way ANOVA or with Student s t-test. P, 0:05 was considered statistically significant. Analyzes were performed with GraphPad Prism, San Diego, USA. 3. Results 3.1. ANP and 8-Br-cGMP preconditioning reduce caspase-3-like activity and caspase-3 processing A maximum of caspase-3 like activity was seen after 24 h of cold ischemia. During reperfusion, caspase-3 like activity decreased to the basal levels of blood-free perfused control livers. Preconditioning with ANP significantly reduced caspase-3 like activity by 53% after 24 h of cold ischemia and by 52% after 45 min of reperfusion in Fig. 1. ANP and 8-Br-cGMP preconditioning reduce caspase-3-like activity. At indicated time points (0 0 : shortly perfused, no ischemia; 24 h isch.: 24 h of cold ischemia; 45 0 rep.: 45 min of reperfusion; rep.: 120 min of reperfusion) control livers and pre-treated livers (ANP 200 nm: panel A, 8-Br-cGMP 50 mm: panel B) were snap frozen and caspase-3-like activity was measured as described under Section 2. Data are expressed as percentage of the highest activity. Columns show mean 6 SEM of two to three independent experiments with four to five rat livers. *P <0.05 versus control, # P<0.05 versus 0 0. the ApopTag w Peroxidase In Situ Apoptosis Detection Kit (Intergen Company, Purchase, New York, USA). 10 high-power fields were counted (1.96 mm 2, approximately 4000 hepatocytes) using a Zeiss Axiolab microscope. The ApopTag w staining results were evaluated in combination with morphological criteria. All histological evaluations were performed in a blinded fashion Histological analysis of liver tissue Paraffin embedded livers were cut into 6 mm sections. HE staining was performed using a standard protocol. Proliferating cells were identified by staining of liver sections with a monoclonal Ki67-antibody (Dianova, dia-5055, Hamburg, Germany), which were pretreated for 30 min with Protex I (Quartett, Berlin, Germany). Ten mid-power fields were counted (approximately 10,000 hepatocytes). Trypan blue uptake of necrotic cells was evaluated at the end of the 2-h reperfusion period. Livers were perfused with trypan blue (0.2 mmol/l) for 10 min and fixed with formalin (1.5%) during a 5-min perfusion. The liver tissue was embedded in paraffin and stained with eosin. Ten high power fields (1.5 mm 2, approximately 3100 hepatocytes) were counted. The percentage of trypan blue stained cells around randomly chosen central and portal veins was determined. All histological evaluations were performed without knowledge from which groups samples were taken from. Fig. 2. Reduced processing of caspase-3 in ANP pre-treated livers. At indicated time points (0 0 : shortly perfused, no ischemia; 24 h isch.: 24 h of cold ischemia; rep.: 120 min of reperfusion) control livers and pre-treated livers (ANP 200 nm) were snap frozen. Western blot with caspase-3 antibody was performed as described under Section 2. CPP32: caspase-3 precursor; p17: proteolytic active subunit. Data show one representative blot out of three independent experiments (mean 6 SEM) with two to three rat livers. Panel A: Western blot; panel B: densitometric analysis of CPP32; panel C: densitometric analysis of p17. *P <0.01 versus control, # P<0.05 versus 0 0.

4 344 T. Gerwig et al. / Journal of Hepatology 39 (2003) Fig. 3. (A) Morphological characteristics of apoptotic cell death. As described in detail under Section 2, apoptotic cell death was determined by biochemical and morphological techniques in order to clearly discriminate from necrotic cell death. The picture shows one representive TUNEL staining after 24 h of cold ischemia. Morphological criteria of apoptosis in TUNEL stained hepatocytes were cell shrinkage, chromatin condensation and margination, and the occurrence of apoptotic bodies. Original magnification: 400-fold. (B) ANP and 8-Br-cGMP preconditioning decrease proportion of apoptotic cells after ischemia. At indicated time points (0 0 : shortly perfused, no ischemia; 24 h isch.: 24 h of cold ischemia; 45 0 rep.: 45 min of reperfusion; rep.: 120 min of reperfusion) control livers and pre-treated livers (ANP 200 nm: panel A, 8-Br-cGMP 50 mm: panel B) were snap frozen and prepared for TUNEL analysis as described under Section 2. Columns show numbers of apoptotic cells (mean 6 SEM, with mean in numerical values) in four to five rat livers. Ten high power fields (1.96 mm 2, approx hepatocytes) were counted at a magnification of 400-fold. *P <0.05 versus control, # P<0.05 versus 0 0. comparison to control livers (Fig. 1, panel A). Perfusion of livers with 8-Br-cGMP, an analogue of the second messenger of ANP, showed a similar significant reduction of caspase-3 like activity after 24 h of cold ischemia. The extent of inhibition after ischemia and 45 min of reperfusion was 51 and 61%, respectively (Fig. 1, panel B). Western blot analysis was performed to detect caspase-3 protein cleavage. In pre-ischemic liver homogenates, caspase-3 precursor CPP32 was clearly detectable in contrast to its cleavage product p17, the proteolytic active subunit. After 24 h of cold ischemia both precursor and proteolytic active subunit increased to a maximum indicating highest enzyme activity. At 120 min of reperfusion, only small amounts of precursor and p17 cleavage product were detectable. Preconditioning with ANP (Fig. 2, panels A, B and C)

5 T. Gerwig et al. / Journal of Hepatology 39 (2003) of TUNEL positive hepatocytes after 24 h of cold ischemia by 37% and 28%, respectively. TUNEL staining of endothelial cells was not affected by ANP nor by 8-Br-cGMP (Fig. 3B) Decreased degenerative changes in ANP pre-treated livers HE staining of cold preserved livers displayed significant evidence of cell injury. Hepatocytes showed large, clear vacuoles within the cytoplasm. Moderate swelling and loosening of intercellular attachments was visible with predominant location in the periportal area. ANP pretreatment decreased degenerative changes with less vacuolization and improved cellular integrity (Fig. 4) ANP preconditioning attenuates necrosis Fig. 4. Degenerative changes of hepatocytes are decreased by ANP pretreatment. After 24 h of preservation in UW solution, liver slices were fixed in formalin solution (1.5%) and embedded in paraffin. HE staining was performed using a standard protocol. (A) Arrows show cytoplasmatic vacuolization as an indicator of degenerative cell damage in a representative control liver. (B) Representative picture of an ANP-pretreated liver (200 nm). Original magnification: 400-fold. diminished the amount of caspase-3 precursor CPP32 and the active p17 subunit compared to control livers after 24 h of cold ischemia ANP and 8-Br-cGMP preconditioning decrease the proportion of TUNEL-positive cells TUNEL staining of liver sections at the beginning of perfusion showed only few TUNEL positive hepatocytes and endothelial cells with characteristic morphology for apoptosis. After 24 h of cold ischemia proportion of TUNEL positive hepatocytes increased significantly. Endothelial cells showed a similar time pattern for TUNEL staining. Apoptotic cells accounted for ^ 0.05% in control livers after 24 h of cold ischemia. Preconditioning with ANP as well as with 8-Br-cGMP reduced the number Histological analysis of livers perfused with trypan blue at the end of the 2-h reperfusion period revealed increased staining of liver cells, demonstrating necrotic cell death. The extent of necrosis was calculated to be 10.9 ^ 2.4% for hepatocytes and 17.2 ^ 2.4% for endothelial cells in periportal liver areas. Pericentrally, 10.3 ^ 1.3% of hepatocytes and 13.1 ^ 1.2% of endothelial cells were necrotic. Pre-treatment with ANP significantly reduced trypan blue uptake in hepatocytes by about 75% in periportal but not in pericentral areas. Similarly, endothelial cells were markedly protected by ANP preconditioning by about 87% in periportal but not in pericentral areas (Fig. 5, panels A and B) No influence of ANP on liver proliferation Histochemical Ki67 antigen staining revealed a large increase of proliferation after cold storage. No changes of proliferation were detected in ANP pre-treated livers compared to control organs (Fig. 6). 4. Discussion To further elucidate the mechanisms of ANP mediated protection against IRI of the isolated perfused rat liver this work specifically investigated necrotic and apoptotic cell death. Our study provides the following novel findings: (1) ANP attenuates apoptotic cell damage. (2) Cold ischemia induces procaspase-3. (3) ANP reduces ischemic degenerative cell changes. (4) During reperfusion, necrotic cell damage is decreased by ANP predominantly in periportal areas. (5) Liver proliferation is not affected by ANP pretreatment. (6) In our model of IRI in isolated perfused rat liver necrotic cell damage prevails over apoptosis.

6 346 T. Gerwig et al. / Journal of Hepatology 39 (2003) Mode of cell death in this model of cold liver ischemia and reperfusion Fig. 5. Necrotic damage during reperfusion is reduced by ANP preconditioning in hepatocytes. After 24 h of preservation in UW solution and 2 h of reperfusion with KH buffer, livers were perfused with trypan blue (0.2 mmol/l) for 10 min and fixed with formalin (1.5%) during a 5-min perfusion as described under Section 2. Panel A: percentage of necrotic hepatocytes in periportal and pericentral areas, panel B: percentage of necrotic endothelial cells. Columns show the percentage of trypan blue stained cells around randomly selected central and portal veins. Magnification 400-fold (mean 6 SEM of four to five different rat livers). *P <0.001 versus control. Fig. 6. No influence of ANP on liver proliferation. At indicated time points (0 0 : shortly perfused, no ischemia; 24 h isch.: 24 h of cold ischemia; 45 0 rep.: 45 min of reperfusion; rep.: 120 min of reperfusion) control livers and pre-treated liver slices (ANP 200 nm) were fixed in formalin solution (1.5%) and embedded in paraffin. Histochemical detection of the proliferation marker Ki67 was performed as described under Section 2. Data (mean 6 SEM) show percentage of Ki67-positive cells per 10 midpower fields (approximately 10,000 hepatocytes). Four to five different rat livers were counted. There were no significant differences versus untreated control livers. # P<0.01 versus 0 0. We evaluated necrotic and apoptotic cell death by common biochemical and histological techniques [26], and regarding zonal and cellular aspects. In addition, morphological criteria were taken into account for a detailed identification of the respective kind of cell death [21]: cells had to show characteristic cell shrinkage, chromatin condensation and margination, and the occurrence of apoptotic bodies to be defined as apoptotic. Furthermore, conditions of the TUNEL assay were chosen with no background staining. In view of increasing controversy over the role of apoptosis in hepatic IRI [21], we aimed at unequivocal identification of apoptotic cells. Cells were counted as necrotic when showing increased eosinophilia, vacuolization, cell disruption, loss of architecture and karyolysis. These investigations were performed in a blinded fashion. Apoptotic cell death was highest after 24 h of cold ischemia. This demonstrates an activation of the apoptotic cascade already during the ischemic period supporting the observations of Rentsch et al. [30]. TUNEL analysis revealed that mainly hepatocytes were affected, whereas SEC showed only minor damage. Our finding is in contrast to other groups, who regard SEC as the main target of reperfusion injury [31], but results similar to ours were also found by others [30]. Apoptotic cell death comprised about 0.8% of total cells, less than one tenth of the proportion of necrotic cells. This points to a minor relevance of apoptosis in our model of IRI [22]. Again, our findings are in contrast to Claviens group [19]. This discrepancy could be due to differences between isolated perfused livers and in vivo experiments. However, also recent in-vivo observations support a minor role of apoptosis [22]. Finally, we decided to perform our study with the experimental conditions previously used to describe hepatoprotection by ANP. This model of isolated perfusion is not affected by systemic effects and can thus focus on intrahepatic mechanisms. To evaluate necrosis, trypan blue uptake into damaged cells was investigated. This dye is selectively taken up by injured cells and is commonly regarded as an indicator of necrotic cell death [32]. After 2 h of reperfusion both periportal and pericentral regions of trypan blue perfused livers showed necrotic changes in hepatocytes and SEC. The evaluation of the degree of necrotic cell death suggests its dominant role in liver injury: about 11% of hepatocytes and 17% of endothelial cells were damaged and therefore necrosis seems to be more relevant than apoptotic damage in this experimental model Reduction of apoptosis by ANP treatment Preconditioning with ANP led to a significant reduction of apoptosis that was most prominent after 24 h of cold

7 T. Gerwig et al. / Journal of Hepatology 39 (2003) ischemia. Protection affected predominantly hepatocytes, whereas apoptosis of SEC was not influenced by ANP. Interestingly, preconditioning of livers with 8-Br-cGMP resulted in a similar reduction of apoptosis. This analogue of ANP s second messenger cgmp has previously been shown to mimic ANP action on liver protection, suggesting a cgmp-mediated mechanism [24]. Preconditioning with either compound reduced apoptotic cell death after 24 h of ischemia by half Caspase-3 upregulation during cold ischemia Interestingly, Western blot analysis revealed a significant increase of caspase-3 precursor CPP32 during the ischemic period. Recently, a transcriptional [33] and translational [34] caspase-3 induction was demonstrated in a rat model of focal cerebral ischemia. The occurrence of such energydependent processes at 48C may be surprising. Indeed, the possibility of transcriptional induction in cold ischemia has been shown for other genes before: despite substrate depletion and reduced metabolism, endothelin-1 is upregulated in the preserved kidney [35]. In addition, there are clear evidences for metabolic activity in cold ischemia as demonstrated by stimulation of stress-activated protein kinases [36,37]. Surprisingly, the amount of caspase-3 precursor CPP32 declined during reperfusion period. This might be explained by the non-recirculating perfusion system employed, leading to removal of apoptotic cells by wash-out ANP reduces early degenerative cell changes HE staining of ischemic livers before reperfusion did not identify necrotic cells, but revealed loss of cell integrity and increased vacuolization. These reversible, predominantly periportal changes seem to progress necrotic cell death in the reperfusion period. ANP revealed its protective potential already after 24 h of cold ischemia by clearly reducing these features of cell degeneration Zonal distribution of hepatoprotection Preconditioning with ANP markedly reduced necrotic damage after 2 h of reperfusion. This protection may be mediated by the property of ANP to prevent cell damage caused by activated Kupffer cells (KC) [38]. Indeed, the zonal pattern of protection seems to depend on the regional distribution of Kupffer cells: these macrophages are predominantly localized in periportal liver areas and periportal KC show higher activity than KC from perivenous areas [39,40]. These findings can explain the zonal distribution of ANP mediated protection No influence of ANP on liver proliferation After orthotopic liver transplantation regenerative pathways are initiated in order to compensate the injury sustained from ischemia and reperfusion [41]. As yet, there is little information about proliferation after cold ischemia and during early reperfusion. We have observed a marked increase of proliferation after ischemia, accounting for 3 5% of hepatocytes. ANP preconditioning did not influence the proliferative response of the organ. Effects at intervals of reperfusion longer than investigated here cannot be excluded by our experiments. The finding that ANP reduces apoptotic together with necrotic cell death in the liver represents a novel aspect of ANP action. ANP has been reported to induce apoptosis in cardiac myocytes of rats [42], whereas ANP protected rat PC12 cells from apoptosis [43]. This suggests a highly celltype specific regulation of apoptotic processes exerted by ANP. Regarding hepatocytes induction of p38 mitogenactivated protein kinase (MAPK) attenuates apoptotic cell death [44]. Interestingly, we recently demonstrated a strong increase of p38 MAPK activity by preconditioning rat livers with ANP [37]. Furthermore, induction of heat shock protein hsp70, which suppresses apoptosis in hepatocytes [45] is another consequence of ANP preconditioning [25]. In conclusion, ANP preconditioning leads to a marked protection against hepatic IRI. Interestingly, apoptosis as well as necrosis are affected making this compound unique in modulating different modes of cell death. In isolated perfused liver, ANP protects most prominently against necrosis of hepatocytes and endothelial cells in periportal zones. Acknowledgements The excellent technical assistance of Ingrid Lib and Andrea Sendelhofert is gratefully acknowledged. This work was supported by the Deutsche Forschungsgemeinschaft (DFG: Ge 576/14-2 and FOR 440/1, TP2). References [1] Bilzer M, Gerbes AL. Preservation injury of the liver: mechanisms and novel therapeutic strategies. J Hepatol 2000;32: [2] Rosser BG, Gores GJ. Liver cell necrosis: cellular mechanisms and clinical implications. Gastroenterology 1995;108: [3] Takei Y, Gao WS, Hijioka T, Savier E, Lindert KA, Lemasters JJ, et al. Increase in survival of liver grafts after rinsing with warm Ringer s solution due to improvement of hepatic microcirculation. Transplantation 1991;52: [4] Upadhya AG, Harvey RP, Howard TK, Lowell JA, Shenoy S, Strasberg SM. Evidence of a role for matrix metalloproteinases in cold preservation injury of the liver in humans and in the rat. Hepatology 1997;26: [5] Calmus Y, Cynober L, Dousset B, Lim SK, Soubrane O, Conti F, et al. Evidence for the detrimental role of proteolysis during liver preservation in humans. Gastroenterology 1995;108: [6] Hansen TN, Dawson PE, Brockbank KG. Effects of hypothermia upon

8 348 T. Gerwig et al. / Journal of Hepatology 39 (2003) endothelial cells: mechanisms and clinical importance. Cryobiology 1994;31: [7] Marsh DC, Lindell SL, Fox LE, Belzer FO, Southard JH. Hypothermic preservation of hepatocytes. I. Role of cell swelling. Cryobiology 1989;26: [8] Otto G, Wolff H, David H. Preservation damage in liver transplantation: electron-microscopic findings. Transplant Proc 1984;16: [9] McKeown CM, Edwards V, Phillips MJ, Harvey PR, Petrunka CN, Strasberg SM. Sinusoidal lining cell damage: the critical injury in cold preservation of liver allografts in the rat. Transplantation 1988;46: [10] Mochida S, Arai M, Ohno A, Masaki N, Ogata I, Fujiwara K. Oxidative stress in hepatocytes and stimulatory state of Kupffer cells after reperfusion differ between warm and cold ischemia in rats. Liver 1994;14: [11] Wanner GA, Ertel W, Muller P, Hofer Y, Leiderer R, Menger MD, et al. Liver ischemia and reperfusion induces a systemic inflammatory response through Kupffer cell activation. Shock 1996;5: [12] Zhou W, McCollum MO, Levine BA, Olson MS. Inflammation and platelet-activating factor production during hepatic ischemia/ reperfusion. Hepatology 1992;16: [13] Le Moine O, Louis H, Stordeur P, Collet JM, Goldman M, Deviere J. Role of reactive oxygen intermediates in interleukin 10 release after cold liver ischemia and reperfusion in mice. Gastroenterology 1997; 113: [14] Kiemer AK, Vollmar AM, Bilzer M, Gerwig T, Gerbes AL. Atrial natriuretic peptide reduces expression of TNF-alpha mrna during reperfusion of the rat liver upon decreased activation of NF-kappaB and AP-1. J Hepatol 2000;33: [15] Jaeschke H, Smith CW, Clemens MG, Ganey PE, Roth RA. Mechanisms of inflammatory liver injury: adhesion molecules and cytotoxicity of neutrophils. Toxicol Appl Pharmacol 1996;139: [16] Gottlieb RA, Burleson KO, Kloner RA, Babior BM, Engler RL. Reperfusion injury induces apoptosis in rabbit cardiomyocytes. J Clin Invest 1994;94: [17] Edwards AD, Mehmet H. Apoptosis in perinatal hypoxic-ischaemic cerebral damage. Neuropathol Appl Neurobiol 1996;22: [18] Leist M, Jaattela M. Four deaths and a funeral: from caspases to alternative mechanisms. Nat Rev Mol Cell Biol 2001;2: [19] Gao W, Bentley RC, Madden JF, Clavien PA. Apoptosis of sinusoidal endothelial cells is a critical mechanism of preservation injury in rat liver transplantation. Hepatology 1998;27: [20] Kohli V, Selzner M, Madden JF, Bentley RC, Clavien PA. Endothelial cell and hepatocyte deaths occur by apoptosis after ischemiareperfusion injury in the rat liver. Transplantation 1999;67: [21] Gujral JS, Bucci TJ, Farhood A, Jaeschke H. Mechanism of cell death during warm hepatic ischemia-reperfusion in rats: apoptosis or necrosis? Hepatology 2001;33: [22] Redaelli CA, Tian YH, Schaffner T, Ledermann M, Baer HU, Dufour JF. Extended preservation of rat liver graft by induction of heme oxygenase-1. Hepatology 2002;35: [23] Bilzer M, Witthaut R, Paumgartner G, Gerbes AL. Prevention of ischemia/reperfusion injury in the rat liver by atrial natriuretic peptide. Gastroenterology 1994;106: [24] Gerbes AL, Vollmar AM, Kiemer AK, Bilzer M. The guanylate cyclase-coupled natriuretic peptide receptor: a new target for prevention of cold ischemia-reperfusion damage of the rat liver. Hepatology 1998;28: [25] Kiemer AK, Gerbes AL, Bilzer M, Vollmar AM. The atrial natriuretic peptide and cgmp: novel activators of the heat shock response in rat livers. Hepatology 2002;35: [26] Miyoshi H, Rust C, Roberts PJ, Burgart LJ, Gores GJ. Hepatocyte apoptosis after bile duct ligation in the mouse involves Fas. Gastroenterology 1999;117: [27] Nicholson DW, Ali A, Thornberry NA, Vaillancourt JP, Ding CK, Gallant M, et al. Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature 1995;376: [28] Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193: [29] Gavrieli Y, Sherman Y, Ben Sasson SA. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 1992;119: [30] Rentsch M, Beham A, Iesalnieks I, Mirwald T, Anthuber M, Jauch KW. Impact of prolonged cold ischemia and reperfusion on apoptosis, activation of caspase 3, and expression of bax after liver transplantation in the rat. Transplant Proc 2001;33: [31] Natori S, Selzner M, Valentino KL, Fritz LC, Srinivasan A, Clavien PA, et al. Apoptosis of sinusoidal endothelial cells occurs during liver preservation injury by a caspase-dependent mechanism. Transplantation 1999;68: [32] Belinsky SA, Popp JA, Kauffman FC, Thurman RG. Trypan blue uptake as a new method to investigate hepatotoxicity in periportal and pericentral regions of the liver lobule: studies with allyl alcohol in the perfused liver. J Pharmacol Exp Ther 1984;230: [33] Harrison DC, Davis RP, Bond BC, Campbell CA, James MF, Parsons AA, et al. Caspase mrna expression in a rat model of focal cerebral ischemia. Brain Res Mol Brain Res 2001;89: [34] Krupinski J, Lopez E, Marti E, Ferrer I. Expression of caspases and their substrates in the rat model of focal cerebral ischemia. Neurobiol Dis 2000;7: [35] Wilhelm SM, Simonson MS, Robinson AV, Stowe NT, Schulak JA. Cold ischemia induces endothelin gene upregulation in the preserved kidney. J Surg Res 1999;85: [36] Crenesse D, Gugenheim J, Hornoy J, Tornieri K, Laurens M, Cambien B, et al. Protein kinase activation by warm and cold hypoxiareoxygenation in primary-cultured rat hepatocytes-jnk(1)/sapk(1) involvement in apoptosis. Hepatology 2000;32: [37] Kiemer AK, Heinze SK, Gerwig T, Gerbes AL, Vollmar AM. Stimulation of p38 MAPK by hormonal preconditioning with atrial natriuretic peptide. World J Gastroenterol 2002;8: [38] Bilzer M, Jaeschke H, Vollmar AM, Paumgartner G, Gerbes AL. Prevention of Kupffer cell-induced oxidant injury in rat liver by atrial natriuretic peptide. Am J Physiol 1999;276:G1137 G1144. [39] Vollmar B, Glasz J, Post S, Menger MD. Depressed phagocytic activity of Kupffer cells after warm ischemia-reperfusion of the liver. J Hepatol 1994;20: [40] Sleyster EC, Knook DL. Relation between localization and function of rat liver Kupffer cells. Lab Invest 1982;47: [41] Olthoff KM. Molecular pathways of regeneration and repair after liver transplantation. World J Surg 2002;26: [42] Filippatos GS, Gangopadhyay N, Lalude O, Parameswaran N, Said SI, Spielman W, et al. Regulation of apoptosis by vasoactive peptides. Am J Physiol Lung Cell Mol Physiol 2001;281:L749 L761. [43] Fiscus RR, Tu AW, Chew SB. Natriuretic peptides inhibit apoptosis and prolong the survival of serum-deprived PC12 cells. Neuroreport 2001;12: [44] Roberts RA, James NH, Cosulich SC. The role of protein kinase B and mitogen-activated protein kinase in epidermal growth factor and tumor necrosis factor alpha-mediated rat hepatocyte survival and apoptosis. Hepatology 2000;31: [45] Ikeyama S, Kusumoto K, Miyake H, Rokutan K, Tashiro S. A nontoxic heat shock protein 70 inducer, geranylgeranylacetone, suppresses apoptosis of cultured rat hepatocytes caused by hydrogen peroxide and ethanol. J Hepatol 2001;35:53 61.

Expression of acid base transporters in the kidney collecting duct in Slc2a7 -/-

Expression of acid base transporters in the kidney collecting duct in Slc2a7 -/- Supplemental Material Results. Expression of acid base transporters in the kidney collecting duct in Slc2a7 -/- and Slc2a7 -/- mice. The expression of AE1 in the kidney was examined in Slc26a7 KO mice.

More information

Focus Application. Compound-Induced Cytotoxicity

Focus Application. Compound-Induced Cytotoxicity xcelligence System Real-Time Cell Analyzer Focus Application Compound-Induced Cytotoxicity Featured Study: Using the Time Resolving Function of the xcelligence System to Optimize Endpoint Viability and

More information

Western Immunoblotting Preparation of Samples:

Western Immunoblotting Preparation of Samples: Western Immunoblotting Preparation of Samples: Total Protein Extraction from Culture Cells: Take off the medium Wash culture with 1 x PBS 1 ml hot Cell-lysis Solution into T75 flask Scrap out the cells

More information

number Done by Corrected by Doctor Heyam Awad

number Done by Corrected by Doctor Heyam Awad number 4 Done by Waseem Abu Obeida Corrected by Saad Al-Hayek Doctor Heyam Awad Cell injury -in the previous lectures we talked about the causes (etiology) and the mechanism (pathogenesis) of cell injury.

More information

Focus Application. Compound-Induced Cytotoxicity

Focus Application. Compound-Induced Cytotoxicity xcelligence System Real-Time Cell Analyzer Focus Application Compound-Induced Cytotoxicity For life science research only. Not for use in diagnostic procedures. Featured Study: Using the Time Resolving

More information

(A) PCR primers (arrows) designed to distinguish wild type (P1+P2), targeted (P1+P2) and excised (P1+P3)14-

(A) PCR primers (arrows) designed to distinguish wild type (P1+P2), targeted (P1+P2) and excised (P1+P3)14- 1 Supplemental Figure Legends Figure S1. Mammary tumors of ErbB2 KI mice with 14-3-3σ ablation have elevated ErbB2 transcript levels and cell proliferation (A) PCR primers (arrows) designed to distinguish

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION FOR Liver X Receptor α mediates hepatic triglyceride accumulation through upregulation of G0/G1 Switch Gene 2 (G0S2) expression I: SUPPLEMENTARY METHODS II: SUPPLEMENTARY FIGURES

More information

Islet viability assay and Glucose Stimulated Insulin Secretion assay RT-PCR and Western Blot

Islet viability assay and Glucose Stimulated Insulin Secretion assay RT-PCR and Western Blot Islet viability assay and Glucose Stimulated Insulin Secretion assay Islet cell viability was determined by colorimetric (3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide assay using CellTiter

More information

Cathepsin K Activity Assay Kit

Cathepsin K Activity Assay Kit Cathepsin K Activity Assay Kit Catalog Number KA0769 100 assays Version: 03 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 General Information... 4 Materials

More information

PRODUCT INFORMATION & MANUAL

PRODUCT INFORMATION & MANUAL PRODUCT INFORMATION & MANUAL Nuclear Extraction Kit NBP2-29447 Research use only. Not for diagnostic or therapeutic procedures. www.novusbio.com - P: 888.506.6887 - technical@novusbio.com Novus kits are

More information

SUPPLEMENTAL MATERIAL. Supplementary Methods

SUPPLEMENTAL MATERIAL. Supplementary Methods SUPPLEMENTAL MATERIAL Supplementary Methods Culture of cardiomyocytes, fibroblasts and cardiac microvascular endothelial cells The isolation and culturing of neonatal rat ventricular cardiomyocytes was

More information

Supplementary material: Materials and suppliers

Supplementary material: Materials and suppliers Supplementary material: Materials and suppliers Electrophoresis consumables including tris-glycine, acrylamide, SDS buffer and Coomassie Brilliant Blue G-2 dye (CBB) were purchased from Ameresco (Solon,

More information

Impact factor: Reporter:4A1H0019 Chen Zi Hao 4A1H0023 Huang Wan ting 4A1H0039 Sue Yi Zhu 4A1H0070 Lin Guan cheng 4A1H0077 Chen Bo xuan

Impact factor: Reporter:4A1H0019 Chen Zi Hao 4A1H0023 Huang Wan ting 4A1H0039 Sue Yi Zhu 4A1H0070 Lin Guan cheng 4A1H0077 Chen Bo xuan Curcumin Protects Neonatal Rat Cardiomyocytes against High Glucose-Induced Apoptosis via PI3K/Akt Signalling Pathway Wei Yu,1,2 Wenliang Zha,1 Zhiqiang Ke,1 Qing Min,2 Cairong Li,1 Huirong Sun,3 and Chao

More information

Supporting Information

Supporting Information Supporting Information Pang et al. 10.1073/pnas.1322009111 SI Materials and Methods ELISAs. These assays were performed as previously described (1). ELISA plates (MaxiSorp Nunc; Thermo Fisher Scientific)

More information

20S Proteasome Activity Assay Kit

20S Proteasome Activity Assay Kit 20S Proteasome Activity Assay Kit For 100 Assays Cat. No. APT280 FOR RESEARCH USE ONLY NOT FOR USE IN DIAGNOSTIC PROCEDURES USA & Canada Phone: +1(800) 437-7500 Fax: +1 (951) 676-9209 Europe +44 (0) 23

More information

HCC1937 is the HCC1937-pcDNA3 cell line, which was derived from a breast cancer with a mutation

HCC1937 is the HCC1937-pcDNA3 cell line, which was derived from a breast cancer with a mutation SUPPLEMENTARY INFORMATION Materials and Methods Human cell lines and culture conditions HCC1937 is the HCC1937-pcDNA3 cell line, which was derived from a breast cancer with a mutation in exon 20 of BRCA1

More information

Caspase-3 Assay Cat. No. 8228, 100 tests. Introduction

Caspase-3 Assay Cat. No. 8228, 100 tests. Introduction Introduction Caspase-3 Assay Cat. No. 8228, 100 tests Caspase-3 is a member of caspases that plays a key role in mediating apoptosis, or programmed cell death. Upon activation, it cleaves a variety of

More information

Cellular responses to stress

Cellular responses to stress Cellular responses to stress (Adaptations, injury and death) (2 of 5) Most injurious stimuli are grouped into: Oxygen deprivation Chemical agents Infectious agents Immunologic reactions Genetic factors

More information

Introduction to pathology lecture 5/ Cell injury apoptosis. Dr H Awad 2017/18

Introduction to pathology lecture 5/ Cell injury apoptosis. Dr H Awad 2017/18 Introduction to pathology lecture 5/ Cell injury apoptosis Dr H Awad 2017/18 Apoptosis = programmed cell death = cell suicide= individual cell death Apoptosis cell death induced by a tightly regulated

More information

Anti-Apoptotic Effects of Cellular Therapy

Anti-Apoptotic Effects of Cellular Therapy Anti-Apoptotic Effects of Cellular Therapy Jason Lapetoda 1, Lee K Landeen, PhD 1, George K Michalopoulos, MD 2, Patricia W Bedard, PhD 1 1 Vital Therapies, Inc., San Diego, CA, USA 2 University of Pittsburgh

More information

TFEB-mediated increase in peripheral lysosomes regulates. Store Operated Calcium Entry

TFEB-mediated increase in peripheral lysosomes regulates. Store Operated Calcium Entry TFEB-mediated increase in peripheral lysosomes regulates Store Operated Calcium Entry Luigi Sbano, Massimo Bonora, Saverio Marchi, Federica Baldassari, Diego L. Medina, Andrea Ballabio, Carlotta Giorgi

More information

Supplemental Information

Supplemental Information Electronic Supplementary Material (ESI) for Food & Function. This journal is The Royal Society of Chemistry 2016 Supplemental Information Supplementary Materials and Methods Materials Assay kits of total

More information

Receptor-interacting Protein Kinases Mediate Necroptosis In Neural Tissue Damage After Spinal Cord Injury

Receptor-interacting Protein Kinases Mediate Necroptosis In Neural Tissue Damage After Spinal Cord Injury Receptor-interacting Protein Kinases Mediate Necroptosis In Neural Tissue Damage After Spinal Cord Injury Haruo Kanno, M.D., Ph.D., Hiroshi Ozawa, M.D., Ph.D., Satoshi Tateda, M.D., Kenichiro Yahata, M.D.,

More information

A263 A352 A204. Pan CK. pstat STAT3 pstat3 STAT3 pstat3. Columns Columns 1-6 Positive control. Omentum. Rectosigmoid A195.

A263 A352 A204. Pan CK. pstat STAT3 pstat3 STAT3 pstat3. Columns Columns 1-6 Positive control. Omentum. Rectosigmoid A195. pstat3 75 Pan CK A A263 A352 A24 B Columns 1-6 Positive control A195 A22 A24 A183 Rectal Nodule STAT3 pstat3 STAT3 pstat3 Columns 7-12 Omentum Rectosigmoid Left Ovary Right Ovary Omentum Uterus Uterus

More information

AMR in Liver Transplantation: Incidence

AMR in Liver Transplantation: Incidence AMR in Liver Transplantation: Incidence Primary AMR 1/3 to 1/2 of ABO-incompatible transplants Uncommon with ABO-compatible transplant Secondary AMR Unknown incidence: rarely tested Why is AMR uncommon

More information

Procaspase-3. Cleaved caspase-3. actin. Cytochrome C (10 M) Z-VAD-fmk. Procaspase-3. Cleaved caspase-3. actin. Z-VAD-fmk

Procaspase-3. Cleaved caspase-3. actin. Cytochrome C (10 M) Z-VAD-fmk. Procaspase-3. Cleaved caspase-3. actin. Z-VAD-fmk A HeLa actin - + + - - + Cytochrome C (1 M) Z-VAD-fmk PMN - + + - - + actin Cytochrome C (1 M) Z-VAD-fmk Figure S1. (A) Pan-caspase inhibitor z-vad-fmk inhibits cytochrome c- mediated procaspase-3 cleavage.

More information

PREPARED BY P.DHARANI PRASAD II YEAR B.PHARM II SEM SUB:PATHOPHYSIOLOGY

PREPARED BY P.DHARANI PRASAD II YEAR B.PHARM II SEM SUB:PATHOPHYSIOLOGY CELL INJURY UNIT I PREPARED BY P.DHARANI PRASAD II YEAR B.PHARM II SEM SUB:PATHOPHYSIOLOGY DETECTION OF CELLULAR CHANGES AFTER INJURY BY: LIGHT MICROSCOPY OR GROSS EXAMINATION DETECT CHANGES HOURS TO DAYS

More information

The Need for a PARP in vivo Pharmacodynamic Assay

The Need for a PARP in vivo Pharmacodynamic Assay The Need for a PARP in vivo Pharmacodynamic Assay Jay George, Ph.D. Chief Scientific Officer Trevigen, Inc. Gaithersburg, MD Poly(ADP-ribose) polymerases are promising therapeutic targets. In response

More information

Supplementary Figure 1: Hsp60 / IEC mice are embryonically lethal (A) Light microscopic pictures show mouse embryos at developmental stage E12.

Supplementary Figure 1: Hsp60 / IEC mice are embryonically lethal (A) Light microscopic pictures show mouse embryos at developmental stage E12. Supplementary Figure 1: Hsp60 / IEC mice are embryonically lethal (A) Light microscopic pictures show mouse embryos at developmental stage E12.5 and E13.5 prepared from uteri of dams and subsequently genotyped.

More information

Multi-Parameter Apoptosis Assay Kit

Multi-Parameter Apoptosis Assay Kit Multi-Parameter Apoptosis Assay Kit Catalog Number KA1335 5 x 96 assays Version: 05 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 Principle of the Assay...

More information

Deposited on: 25 July 2011

Deposited on: 25 July 2011 Morton, E., Macrae, I.M., McCabe, C., Brown, S.M. and White, F. (2006) Identification of the growth arrest and DNA damage protein GADD34 in the normal human heart and demonstration of alterations in expression

More information

Cathepsin K Activity Assay Kit (Fluorometric)

Cathepsin K Activity Assay Kit (Fluorometric) ab65303 Cathepsin K Activity Assay Kit (Fluorometric) Instructions for Use For the rapid, sensitive and accurate measurement of Cathepsin K activity in various samples. This product is for research use

More information

Do Histological Features Inform DILI mechanisms?

Do Histological Features Inform DILI mechanisms? Do Histological Features Inform DILI mechanisms? Drug-Induced Liver Injury (DILI) Conference XVII June, 2017 David Kleiner, M.D., Ph.D. NCI/Laboratory of Pathology Disclosures No financial or other conflicts

More information

The Groningen hypothermic liver perfusion system for improved preservation in organ transplantation Plaats, Arjan van der

The Groningen hypothermic liver perfusion system for improved preservation in organ transplantation Plaats, Arjan van der University of Groningen The Groningen hypothermic liver perfusion system for improved preservation in organ transplantation Plaats, Arjan van der IMPORTANT NOTE: You are advised to consult the publisher's

More information

Sestrin2 and BNIP3 (Bcl-2/adenovirus E1B 19kDa-interacting. protein3) regulate autophagy and mitophagy in renal tubular cells in. acute kidney injury

Sestrin2 and BNIP3 (Bcl-2/adenovirus E1B 19kDa-interacting. protein3) regulate autophagy and mitophagy in renal tubular cells in. acute kidney injury Sestrin2 and BNIP3 (Bcl-2/adenovirus E1B 19kDa-interacting protein3) regulate autophagy and mitophagy in renal tubular cells in acute kidney injury by Masayuki Ishihara 1, Madoka Urushido 2, Kazu Hamada

More information

Glutathione Peroxidase Assay Kit

Glutathione Peroxidase Assay Kit Glutathione Peroxidase Assay Kit Catalog Number KA0882 100 assays Version: 04 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 General Information... 4

More information

Reperfusion Injury: How Can We Reduce It?

Reperfusion Injury: How Can We Reduce It? MI/CAD: Practical Question in Management of AMI Patients Reperfusion Injury: How Can We Reduce It? Hyun-Jai Cho, M.D., Ph.D Cardiovascular Center & Department of Internal Medicine Seoul National University

More information

Organ preservation & transplantation: newest insights & perspectives. Ina Jochmans, MD, PhD Abdominal Transplant Surgery KU Leuven, Belgium

Organ preservation & transplantation: newest insights & perspectives. Ina Jochmans, MD, PhD Abdominal Transplant Surgery KU Leuven, Belgium Organ preservation & transplantation: newest insights & perspectives Ina Jochmans, MD, PhD Abdominal Transplant Surgery KU Leuven, Belgium 1. Expand the donor pool 2. Optimal preservation & resuscitation

More information

The liver in poisoning: what can we learn from animal models?

The liver in poisoning: what can we learn from animal models? The liver in poisoning: what can we learn from animal models? Stephan Krähenbühl Clinical Pharmacology & Toxicology University Hospital 4031 Basel/Switzerland Kraehenbuehl@uhbs.ch Outcome and causes of

More information

Chemical Chaperones Mitigate Experimental Asthma By Attenuating Endoplasmic

Chemical Chaperones Mitigate Experimental Asthma By Attenuating Endoplasmic Chemical Chaperones Mitigate Experimental Asthma By Attenuating Endoplasmic Reticulum Stress Lokesh Makhija, BE, Veda Krishnan, MSc, Rakhshinda Rehman, MTech, Samarpana Chakraborty, MSc, Shuvadeep Maity,

More information

Mass Histology Service

Mass Histology Service Mass Histology Service A complete anatomical pathology laboratory www.masshistology.com Telephone: (877) 286-6004 Report on Pathology A Time Course Study of the Local Effects of Intramuscular XXXXXXX Injection

More information

The Biochemistry of apoptosis

The Biochemistry of apoptosis The Biochemistry of apoptosis 1 1 The apoptosis is composed of multiple biochemical events 2 2 Biochemical, cellular, and molecular events in Apoptosis 1. Membrane blebbing; phosphatidyl serine exposure

More information

Total Histone H3 Acetylation Detection Fast Kit (Colorimetric)

Total Histone H3 Acetylation Detection Fast Kit (Colorimetric) Total Histone H3 Acetylation Detection Fast Kit (Colorimetric) Catalog Number KA1538 48 assays Version: 02 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use...

More information

Supplemental Table 1. Primers used for RT-PCR analysis of inflammatory cytokines Gene Primer Sequence

Supplemental Table 1. Primers used for RT-PCR analysis of inflammatory cytokines Gene Primer Sequence Supplemental Table 1. Primers used for RT-PCR analysis of inflammatory cytokines Gene Primer Sequence IL-1α Forward primer 5 -CAAGATGGCCAAAGTTCGTGAC-3' Reverse primer 5 -GTCTCATGAAGTGAGCCATAGC-3 IL-1β

More information

To determine the effect of over-expression and/or ligand activation of. PPAR / on cell cycle, cell lines were cultured as described above until ~80%

To determine the effect of over-expression and/or ligand activation of. PPAR / on cell cycle, cell lines were cultured as described above until ~80% Supplementary Materials and Methods Cell cycle analysis To determine the effect of over-expression and/or ligand activation of PPAR / on cell cycle, cell lines were cultured as described above until ~80%

More information

Prevention of Ischemia-Reperfusion Injury in the Rat Liver by Atrial Natriuretic Peptide: Insights into Mechanisms and Mode of Cell Death

Prevention of Ischemia-Reperfusion Injury in the Rat Liver by Atrial Natriuretic Peptide: Insights into Mechanisms and Mode of Cell Death Dissertation zur Erlangung des Doktorgrades der Fakultät für Chemie und Pharmazie der Ludwig-Maximilians-Universität München Prevention of Ischemia-Reperfusion Injury in the Rat Liver by Atrial Natriuretic

More information

HISTOPL4THOLOG1CAL STUDY OF LIVER

HISTOPL4THOLOG1CAL STUDY OF LIVER HISTOPL4THOLOG1CAL STUDY OF LIVER 6.1 Inti-oduction The structural and functional organization of the liver has been described by hepatic lobule and hepatic acinus models, respectively (Jarvelainen, 2000).

More information

C-Phycocyanin (C-PC) is a n«sjfc&c- waefc-jduble phycobiliprotein. pigment isolated from Spirulina platensis. This water- soluble protein pigment is

C-Phycocyanin (C-PC) is a n«sjfc&c- waefc-jduble phycobiliprotein. pigment isolated from Spirulina platensis. This water- soluble protein pigment is ' ^Summary C-Phycocyanin (C-PC) is a n«sjfc&c- waefc-jduble phycobiliprotein pigment isolated from Spirulina platensis. This water- soluble protein pigment is of greater importance because of its various

More information

c Ischemia (30 min) Reperfusion (8 w) Supplementary Figure bp 300 bp Ischemia (30 min) Reperfusion (4 h) Dox 20 mg/kg i.p.

c Ischemia (30 min) Reperfusion (8 w) Supplementary Figure bp 300 bp Ischemia (30 min) Reperfusion (4 h) Dox 20 mg/kg i.p. a Marker Ripk3 +/ 5 bp 3 bp b Ischemia (3 min) Reperfusion (4 h) d 2 mg/kg i.p. 1 w 5 w Sacrifice for IF size A subset for echocardiography and morphological analysis c Ischemia (3 min) Reperfusion (8

More information

Chapter 1 CELL INJURY CELL DEATH CELL ADAPTATIONS. M.G.Rajanandh, Dept. of Pharmacy Practice, SRM College of Pharmacy, SRM University.

Chapter 1 CELL INJURY CELL DEATH CELL ADAPTATIONS. M.G.Rajanandh, Dept. of Pharmacy Practice, SRM College of Pharmacy, SRM University. Chapter 1 CELL INJURY CELL DEATH CELL ADAPTATIONS M.G.Rajanandh, Dept. of Pharmacy Practice, SRM College of Pharmacy, SRM University. CONCEPTS IN CELL INJURY The clinical signs and symptoms are several

More information

SensoLyte 520 HDAC Activity Assay Kit *Fluorimetric*

SensoLyte 520 HDAC Activity Assay Kit *Fluorimetric* SensoLyte 520 HDAC Activity Assay Kit *Fluorimetric* Catalog # 72084 Kit Size 100 Assays (96-well plate) Optimized Performance: This kit is optimized to detect HDAC activity. Enhanced Value: It provides

More information

Histopathology: Cell necrosis and cytoplasmic accumulations

Histopathology: Cell necrosis and cytoplasmic accumulations Histopathology: Cell necrosis and cytoplasmic accumulations These presentations are to help you identify basic histopathological features. They do not contain the additional factual information that you

More information

Human Obestatin ELISA

Human Obestatin ELISA K-ASSAY Human Obestatin ELISA For the quantitative determination of obestatin in human serum and plasma Cat. No. KT-495 For Research Use Only. 1 Rev. 081309 K-ASSAY PRODUCT INFORMATION Human Obestatin

More information

shehab Moh Tarek ... ManarHajeer

shehab Moh Tarek ... ManarHajeer 3 shehab Moh Tarek... ManarHajeer In the previous lecture we discussed the accumulation of oxygen- derived free radicals as a mechanism of cell injury, we covered their production and their pathologic

More information

Future of liver transplantation

Future of liver transplantation Future of liver transplantation Transplant Atlantic November 4, 2016 Ian Alwayn, MD, PhD The problem: Population is becoming older, more obese and more comorbid Donor population is becoming older, more

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION Figure S1 Treatment with both Sema6D and Plexin-A1 sirnas induces the phenotype essentially identical to that induced by treatment with Sema6D sirna alone or Plexin-A1 sirna alone. (a,b) The cardiac tube

More information

Protocol for Gene Transfection & Western Blotting

Protocol for Gene Transfection & Western Blotting The schedule and the manual of basic techniques for cell culture Advanced Protocol for Gene Transfection & Western Blotting Schedule Day 1 26/07/2008 Transfection Day 3 28/07/2008 Cell lysis Immunoprecipitation

More information

NF-κB p65 (Phospho-Thr254)

NF-κB p65 (Phospho-Thr254) Assay Biotechnology Company www.assaybiotech.com Tel: 1-877-883-7988 Fax: 1-877-610-9758 NF-κB p65 (Phospho-Thr254) Colorimetric Cell-Based ELISA Kit Catalog #: OKAG02015 Please read the provided manual

More information

Bile acids initiate cholestatic liver injury by triggering a hepatic specific inflammatory response. Supplementary Results

Bile acids initiate cholestatic liver injury by triggering a hepatic specific inflammatory response. Supplementary Results Bile acids initiate cholestatic liver injury by triggering a hepatic specific inflammatory response Shi-Ying Cai 1, Xinshou Ouyang 1, Yonglin Chen 1, Carol J. Soroka 1, Juxian Wang 2, Albert Mennone 1,

More information

Lipid Peroxidation Assay

Lipid Peroxidation Assay Package Insert Lipid Peroxidation Assay 96 Wells For Research Use Only v. 1.0 Eagle Biosciences, Inc. 82 Broad Street, Suite 383, Boston, MA 02110 Phone: 866-419-2019 Fax: 617-419-1110 INTRODUCTION Lipid

More information

The Annexin V Apoptosis Assay

The Annexin V Apoptosis Assay The Annexin V Apoptosis Assay Development of the Annexin V Apoptosis Assay: 1990 Andree at al. found that a protein, Vascular Anticoagulant α, bound to phospholipid bilayers in a calcium dependent manner.

More information

Expression and clinical significance of ADAM17 protein in esophageal squamous cell carcinoma

Expression and clinical significance of ADAM17 protein in esophageal squamous cell carcinoma Expression and clinical significance of ADAM17 protein in esophageal squamous cell carcinoma H.B. Liu, Y. Zhu, Q.C. Yang, Y. Shen, X.J. Zhang and H. Chen Department of Pathology First People s Hospital

More information

renoprotection therapy goals 208, 209

renoprotection therapy goals 208, 209 Subject Index Aldosterone, plasminogen activator inhibitor-1 induction 163, 164, 168 Aminopeptidases angiotensin II processing 64 66, 214 diabetic expression 214, 215 Angiotensin I intrarenal compartmentalization

More information

The mounting number of patients awaiting liver. Improvement of Rat Liver Graft Quality by Pifithrin- Mediated Inhibition of Hepatocyte Necrapoptosis

The mounting number of patients awaiting liver. Improvement of Rat Liver Graft Quality by Pifithrin- Mediated Inhibition of Hepatocyte Necrapoptosis Improvement of Rat Liver Graft Quality by Pifithrin- Mediated Inhibition of Hepatocyte Necrapoptosis Amr M. El-Gibaly, 1 Claudia Scheuer, 1 Michael D. Menger, 1 and Brigitte Vollmar 2 Early graft dysfunction

More information

Proteasome Activity Assay Kit

Proteasome Activity Assay Kit Proteasome Activity Assay Kit Catalog Number KA1431 100 assays Version: 03 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 General Information... 4 Materials

More information

Table S1. Sequence of human and mouse primers used for RT-qPCR measurements.

Table S1. Sequence of human and mouse primers used for RT-qPCR measurements. Table S1. Sequence of human and mouse primers used for RT-qPCR measurements. Ca9, carbonic anhydrase IX; Ndrg1, N-myc downstream regulated gene 1; L28, ribosomal protein L28; Hif1a, hypoxia inducible factor

More information

Introduction: 年 Fas signal-mediated apoptosis. PI3K/Akt

Introduction: 年 Fas signal-mediated apoptosis. PI3K/Akt Fas-ligand (CD95-L; Fas-L) Fas (CD95) Fas (apoptosis) 年 了 不 度 Fas Fas-L 力 不 Fas/Fas-L T IL-10Fas/Fas-L 不 年 Fas signal-mediated apoptosis 度降 不 不 力 U-118, HeLa, A549, Huh-7 MCF-7, HepG2. PI3K/Akt FasPI3K/Akt

More information

APOPTOSIS, NECROSIS AND CANCER. Dr. S. P. Pattanayak

APOPTOSIS, NECROSIS AND CANCER. Dr. S. P. Pattanayak APOPTOSIS, NECROSIS AND CANCER Dr. S. P. Pattanayak LEARNING OBJECTIVES At the end of the lecture, students should be able to: Know the importance of cell death. Define various modes of cell death. Identify

More information

Instructions for Use. APO-AB Annexin V-Biotin Apoptosis Detection Kit 100 tests

Instructions for Use. APO-AB Annexin V-Biotin Apoptosis Detection Kit 100 tests 3URGXFW,QIRUPDWLRQ Sigma TACS Annexin V Apoptosis Detection Kits Instructions for Use APO-AB Annexin V-Biotin Apoptosis Detection Kit 100 tests For Research Use Only. Not for use in diagnostic procedures.

More information

Nuclear Extraction Kit

Nuclear Extraction Kit Nuclear Extraction Kit Catalog Number KA1346 50 assays Version: 07 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Principle of the Assay... 3 General Information... 4

More information

Plate-Based Assay Methods for the Assessment of Cellular Health

Plate-Based Assay Methods for the Assessment of Cellular Health Plate-Based Assay Methods for the Assessment of Cellular Health Andrew L. Niles, Senior Research Scientist 2012, Promega Corporation. Biological Outcomes in Cell Culture Treatment -Small molecule -Bio-molecule

More information

Collagenase Assay Kit

Collagenase Assay Kit Collagenase Assay Kit Catalog # 31 and 32 For Research Use Only - Not Human or Therapeutic Use INTRODUCTION Collagenases are members of the matrix metalloproteinase (MMP) family and degrade collagen types

More information

Supplementary Material for

Supplementary Material for Supplementary Material for Parathyroid Hormone Signaling through Low-density-lipoprotein-related Protein 6 Mei Wan, Chaozhe Yang, Jun Li, Xiangwei Wu, Hongling Yuan, Hairong Ma, Xi He, Shuyi Nie, Chenbei

More information

Types of insult - hypoxia

Types of insult - hypoxia Introduction This presentation will be a guide to cell injury and cell death outline causes and pathogenesis of cell injury/death describe the morphological changes of cell injury/death Describe the process

More information

Chromatin IP (Isw2) Fix soln: 11% formaldehyde, 0.1 M NaCl, 1 mm EDTA, 50 mm Hepes-KOH ph 7.6. Freshly prepared. Do not store in glass bottles.

Chromatin IP (Isw2) Fix soln: 11% formaldehyde, 0.1 M NaCl, 1 mm EDTA, 50 mm Hepes-KOH ph 7.6. Freshly prepared. Do not store in glass bottles. Chromatin IP (Isw2) 7/01 Toshi last update: 06/15 Reagents Fix soln: 11% formaldehyde, 0.1 M NaCl, 1 mm EDTA, 50 mm Hepes-KOH ph 7.6. Freshly prepared. Do not store in glass bottles. 2.5 M glycine. TBS:

More information

EpiQuik Total Histone H3 Acetylation Detection Fast Kit (Colorimetric)

EpiQuik Total Histone H3 Acetylation Detection Fast Kit (Colorimetric) EpiQuik Total Histone H3 Acetylation Detection Fast Kit (Colorimetric) Base Catalog # PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE The EpiQuik Total Histone H3 Acetylation Detection Fast Kit (Colorimetric)

More information

ab Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric)

ab Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) ab156064 Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) Instructions for Use For the quantitative measurement of Histone Deacetylase activity in cell lysates This product is for research

More information

Name Animal source Vendor Cat # Dilutions

Name Animal source Vendor Cat # Dilutions Supplementary data Table S1. Primary and Secondary antibody sources Devi et al, TXNIP in mitophagy A. Primary Antibodies Name Animal source Vendor Cat # Dilutions 1. TXNIP mouse MBL KO205-2 1:2000 (WB)

More information

Molecular biology :- Cancer genetics lecture 11

Molecular biology :- Cancer genetics lecture 11 Molecular biology :- Cancer genetics lecture 11 -We have talked about 2 group of genes that is involved in cellular transformation : proto-oncogenes and tumour suppressor genes, and it isn t enough to

More information

SensoLyte 520 Cathepsin K Assay Kit *Fluorimetric*

SensoLyte 520 Cathepsin K Assay Kit *Fluorimetric* SensoLyte 520 Cathepsin K Assay Kit *Fluorimetric* Catalog # 72171 Kit Size 100 Assays (96-well plate) Optimized Performance: This kit is optimized to detect Cathepsin K activity. Enhanced Value: Ample

More information

Use of a camp BRET Sensor to Characterize a Novel Regulation of camp by the Sphingosine-1-phosphate/G 13 Pathway

Use of a camp BRET Sensor to Characterize a Novel Regulation of camp by the Sphingosine-1-phosphate/G 13 Pathway Use of a camp BRET Sensor to Characterize a Novel Regulation of camp by the Sphingosine-1-phosphate/G 13 Pathway SUPPLEMENTAL DATA Characterization of the CAMYEL sensor and calculation of intracellular

More information

Aconitase Enzyme Activity Microplate Assay Kit

Aconitase Enzyme Activity Microplate Assay Kit ab109712 Aconitase Enzyme Activity Microplate Assay Kit Instructions for Use For the quantitative measurement of Aconitase activity in samples from all species This product is for research use only and

More information

Biodegradable Zwitterionic Nanogels with Long. Circulation for Antitumor Drug Delivery

Biodegradable Zwitterionic Nanogels with Long. Circulation for Antitumor Drug Delivery Supporting Information Biodegradable Zwitterionic Nanogels with Long Circulation for Antitumor Drug Delivery Yongzhi Men, Shaojun Peng, Peng Yang, Qin Jiang, Yanhui Zhang, Bin Shen, Pin Dong, *, Zhiqing

More information

A HISTOPATHOLOGICAL STUDY OF ANALGESIC HEPATOTOXICITY IN RABBIT

A HISTOPATHOLOGICAL STUDY OF ANALGESIC HEPATOTOXICITY IN RABBIT A HISTOPATHOLOGICAL STUDY OF ANALGESIC HEPATOTOXICITY IN RABBIT Pages with reference to book, From 41 To 43 M. Ashraf Qamar, S.M. Alam ( Basic Medical Sciences Institute, Jinnah Postgraduate Medical Centre,

More information

Kit for assay of thioredoxin

Kit for assay of thioredoxin FkTRX-02-V2 Kit for assay of thioredoxin The thioredoxin system is the major protein disulfide reductase in cells and comprises thioredoxin, thioredoxin reductase and NADPH (1). Thioredoxin systems are

More information

Ischemia-induced inflammation is increased and satellite-cell activation is decreased in TNFR2/P75 knockout hindlimb ischemia model

Ischemia-induced inflammation is increased and satellite-cell activation is decreased in TNFR2/P75 knockout hindlimb ischemia model Boston University OpenBU Theses & Dissertations http://open.bu.edu Boston University Theses & Dissertations 2014 Ischemia-induced inflammation is increased and satellite-cell activation is decreased in

More information

References Required document for Laboratory Accreditation by the College of American Pathologists.

References Required document for Laboratory Accreditation by the College of American Pathologists. Subject NT-proBNP Cobas e601 Index Number Lab-4011 Section Laboratory Subsection Chemistry Category Departmental Contact Benjamin Michel Last Revised 10/19/2016 References Required document for Laboratory

More information

Collagenase Assay Kit

Collagenase Assay Kit Collagenase Assay Kit Catalog # 31 and 32 For Research Use Only - Not Human or Therapeutic Use INTRODUCTION The collagenases are members of the matrix metalloproteinase (MMP) family and degrade collagen

More information

The Comet and TUNEL Assays

The Comet and TUNEL Assays Sperm DNA: organization, protection and vulnerability from basic science to clinical application Methods to assess the status of the sperm DNA: The Comet and TUNEL Assays Lars Björndahl Centre for Andrology

More information

27 part 2. Laith Abu Shekha. Mamoon Al-qatameen

27 part 2. Laith Abu Shekha. Mamoon Al-qatameen 27 part 2 Laith Abu Shekha Mamoon Al-qatameen Ebaa Alzayadneh In this sheet we will continue talking about second messengers for hormone that can t cross PM. D. Ca +2 as a second messenger: Another second

More information

SUPPLEMENTARY MATERIAL

SUPPLEMENTARY MATERIAL SUPPLEMENTARY MATERIAL Table S1. Primers and fluorescent probes used for qrt-pcr analysis of relative expression levels of PPP family phosphatases. gene name forward primer, 5-3 probe, 5-3 reverse primer,

More information

BRIDGE THE GAP A Human Pathways Approach to Disease Research

BRIDGE THE GAP A Human Pathways Approach to Disease Research BRIDGE THE GAP A Human Pathways Approach to Disease Research BIOMED 21 Brussels December 8 th, 2015 Brigitte Landesmann Systems Toxicology Unit and EURL ECVAM Institute for Health and Consumer Protection

More information

Total Phosphatidic Acid Assay Kit

Total Phosphatidic Acid Assay Kit Product Manual Total Phosphatidic Acid Assay Kit Catalog Number MET- 5019 100 assays FOR RESEARCH USE ONLY Not for use in diagnostic procedures Introduction Phosphatidic Acid (PA) is a critical precursor

More information

Orthotopic liver transplantation has emerged as the

Orthotopic liver transplantation has emerged as the Cold Preservation Warm Reperfusion Perturbs Cytosolic Calcium Ion Homeostasis in Rat Liver Sinusoidal Endothelial Cells Stéphanie Auger, Diane Vallerand, and Pierre Selim Haddad Increases in intracellular

More information

Pepsin Solution ready-to-use

Pepsin Solution ready-to-use SIE HABEN DIE VISION, WIR HABEN DIE SUBSTANZ. Pepsin Solution Single component Pepsin Solution: only one component refrigerator stable Pepsin is a commonly used digestive enzyme for immunohistochemical

More information

Supplementary Figure 1. Confocal immunofluorescence showing mitochondrial translocation of Drp1. Cardiomyocytes treated with H 2 O 2 were prestained

Supplementary Figure 1. Confocal immunofluorescence showing mitochondrial translocation of Drp1. Cardiomyocytes treated with H 2 O 2 were prestained Supplementary Figure 1. Confocal immunofluorescence showing mitochondrial translocation of Drp1. Cardiomyocytes treated with H 2 O 2 were prestained with MitoTracker (red), then were immunostained with

More information

Secondary Negative Effects of Isolation Enzyme (s) On Human Islets. A.N.Balamurugan

Secondary Negative Effects of Isolation Enzyme (s) On Human Islets. A.N.Balamurugan Secondary Negative Effects of Isolation Enzyme (s) On Human Islets A.N.Balamurugan Human Islets Functional Mass Preservation 18-25 min 10 min 8-12 min 10-15 min 45-60 min pancreas in chamber 37º Sub-Optimal

More information

PUBLICATIONS. cells. J. Physiol. (London) 517P:91P (Manchester, England, UK).

PUBLICATIONS. cells. J. Physiol. (London) 517P:91P (Manchester, England, UK). 277 PUBLICATIONS Abstracts Haddad JJ, Land SC (1999). Differential activation of oxygen-responsive transcription factors over fetal-to-neonatal alveolar oxygen tensions in rat fetal distal lung epithelial

More information

Human IL-2. Pre-Coated ELISA Kit

Human IL-2. Pre-Coated ELISA Kit Human IL-2 (Interleukin 2) Pre-Coated ELISA Kit Catalog No: 90-2083 1 96 well Format (96 tests) Detection Range: 31.2 2000 pg/ml Sensitivity: < 18.75 pg/ml This immunoassay kit allows for the in vitro

More information

Data sheet. TBARS Assay kit. (Colorimetric/Fluorometric) Kit Contents. MDA-TBA Adduct. 2-Thiobarbituric Acid. Cat. No: CA995.

Data sheet. TBARS Assay kit. (Colorimetric/Fluorometric) Kit Contents. MDA-TBA Adduct. 2-Thiobarbituric Acid. Cat. No: CA995. Data sheet Cat. No: CA995 TBARS Assay kit (Colorimetric/Fluorometric) Introduction Oxidative stress in the cellular environment results in the formation of highly reactive and unstable lipid hydroperoxides.

More information