Interictal metabolic changes in episodic migraine: a voxel-based FDG-PET study.

Whereas there are many H(2)(15)O-positron emission tomography (PET) studies demonstrating neuronal activation during acute migraine attacks, little information is available on the interictal (headache-free period) glucose metabolic changes in migraine. We therefore conducted voxel-based statistical parametric mapping analysis of (18)F-fluorodeoxyglucose-PET to evaluate interictal metabolic differences between 20 episodic migraine patients (four with aura; three men; mean age 34.0 +/- 6.4 years) and 20 control subjects. Separate correlation analyses were performed to delineate a possible relationship between regional glucose metabolism and disease duration or lifetime headache frequency in migraine patients. Group comparison showed that migraine patients had significant hypometabolism in several regions known to be involved in central pain processing, such as bilateral insula, bilateral anterior and posterior cingulate cortex, left premotor and prefrontal cortex, and left primary somatosensory cortex (uncorrected P < 0.001, corrected P < 0.05 with small volume corrections). Correlation analyses showed that regional metabolism of the insula and anterior cingulate cortex had significant negative correlations with disease duration and lifetime headache frequency (uncorrected P < 0.001, corrected P < 0.05 with small volume corrections). Our findings of progressive glucose hypometabolism in relation to increasing disease duration and increasing headache frequency suggest that repeated migraine attacks over time lead to metabolic abnormalities of selective brain regions belonging to the central pain matrix.

The diagnostic value of the sagittal multiplanar reconstruction CT images for nasal bone fractures.

AIM: To compare the diagnostic performance of sagittal multiplanar reconstruction (MPR) images and axial images for the detection of a nasal bone fracture. MATERIALS AND METHODS: This prospective study included 533 consecutive patients who underwent three-dimensional images with 64-section multidetector-row CT for the evaluation of a facial bone fracture between June 2007 and May 2008 (366 males; 167 females; mean age +/- standard deviation 31.1+/-21.2 years; age range 1-92 years). Two observers independently scored the possibility of a nasal bone fracture on axial and sagittal images. Receiver operating characteristic (ROC) curve analysis was performed. RESULTS: The Az values of the sagittal images were higher than those of the axial images for both observers (p=0.002 and 0.010, respectively) with higher accuracy (p<0.001 and 0.016, respectively). The sensitivities of sagittal images were superior to those of axial images, especially for type 1simple nasal bone fractures with no or minimal displacement (observer 1, 98.6 versus 72.8%; observer 2, 84.9 versus 71%). CONCLUSION: Sagittal MPR facial bone CT images provided superior diagnostic performance, and their addition to axial images is useful for the evaluation of nasal bone fractures.

Regional grey matter changes in patients with migraine: a voxel-based morphometry study.

We used voxel-based morphometry (VBM) to compare grey matter volume (GMV) between 20 migraine patients (five with aura and 15 without aura) with normal conventional magnetic resonance imaging findings and 33 healthy controls matched for age and sex. A separate analysis was also performed to delineate a possible correlation between the GMV changes and the headache duration or lifetime headache frequency. When compared with controls, migraine patients had significant GMV reductions in the bilateral insula, motor/premotor, prefrontal, cingulate cortex, right posterior parietal cortex, and orbitofrontal cortex (P < 0.001, uncorrected for multiple comparisons at a voxel level; corrected P < 0.05 after small volume corrections). All regions of the GMV changes were negatively correlated with headache duration and lifetime headache frequency (P < 0.05, Pearson's correlation test). We found evidence for structural grey matter changes in patients with migraine. Our findings of progressive GMV reductions in relation to increasing headache duration and increasing headache frequency suggest that repeated migraine attacks over time result in selective damage to several brain regions involved in central pain processing.

Effects of tacrolimus on antioxidant status and oxidative stress in glioma cells.

OBJECTIVES: After organ transplantation, some patients suffer from mild neurologic symptoms, ranging from tremor to severe complications, including seizures and encephalopathy. Among the immunosuppressants, tacrolimus can cause neurologic side effects. However, the mechanisms of encephalopathy by tacrolimus are not fully understood. We measured the antioxidant status, hydrogen peroxide level, and malondialdehyde level in glioma cells after tacrolimus treatment. METHODS: The production of hydrogen peroxide was determined by the modified xylenol orange method. The amount of malondialdehyde was measured by the thiobarbituric acid assay, which is based on malondialdehyde reaction with thiobarbituric acid to give a red species absorbing at 535 nm. Total antioxidant status (TAS) was measured using TAS kits (NX2332). RESULTS: Tacrolimus resulted in dose- and time-dependent increases in the production of hydrogen peroxide by glioma cells. The antioxidant status decreased in the glioma cells after tacrolimus treatment. Malondialdehyde level was unchanged in the glioma cells after tacrolimus treatment. CONCLUSIONS: Increased production of reactive oxygen species and decreased antioxidant status by tacrolimus in glioma cells may contribute to neurologic side effects.

The production of reactive oxygen species in tacrolimus-treated glial cells.

OBJECTIVE: After organ transplantation, some patients suffer from mild neurological symptoms, such as tremor, to severe complications, including seizures and encephalopathy. These neurological side effects can be caused by immunosuppressants such as tacrolimus. However, the mechanism of encephalopathy by tacrolimus is not fully understood. METHODS: We measured the production of reactive oxygen species (ROS) in glioma cells after tacrolimus treatment. Tacrolimus added to glioma cells was incubated for 60 minutes at 37 degrees C. The production of ROS was evaluated by measuring the fluorescent product from the oxidation of an oxidant-sensitive 2',7'-dichlorofluorescin using VICTOR3TM multilabel counter. RESULTS: Tacrolimus resulted in the production of the ROS in glioma cells. The production of the ROS was increased in time-dependent fashion. CONCLUSIONS: These findings indicated that the tacrolimus may contribute the neurological side effects by ROS production.

Cytokine array after cyclosporine treatment in rats.

OBJECTIVES: Long-term treatment with cyclosporine (CsA) results in chronic nephrotoxicity, which is known to be mediated by several cytokines including transforming growth factor-betal. Cytokines are known to play an important role in innate immunity, apoptosis, angiogenesis, cell growth, and differentiation. They are known to be involved in most disease processes, including cancer, cardiac disease, and nephrotoxicity. To evaluate changes of cytokines in a rat model of CsA-induced chronic nephrotoxicity, we performed a cytokine array. METHODS: Experiments were performed on two groups of rats; normal control group and CsA-treated group. Cytokine array in rat serum was performed using Cytokine Antibody Array I kit from RayBiotech. RESULTS: Serum creatinine, urine creatinine, and creatinine clearance increased in the CsA-treated group. Among the several cytokines, the expressions of the lipopolysaccharide-induced CXC chemokine (LIX), monocyte chemoattractant protein 1 (MCP-1), nerve growth factor (beta-NGF), and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the CsA-treated group were increased above that of cytokines in the control group. The density of the LIX in controls was 0.62, and in the CsA-treated group was 1.24. The density of the MCP-1 in controls was 0.68, and in CsA-treated, 1.43. The density of the beta-NGF in controls was 0.62, and that in CsA-treated, 1.24. The density of the TIMP-1 in controls 1.13, and in CsA-treated, 1.40. CONCLUSIONS: Our data suggested that among several cytokines elevated levels of the LIX, MCP-1, beta-NGF, and TIMP-1 are the contributing factors to CsA-induced nephropathy.

MR Imaging for Differentiating Contrast Staining from Hemorrhagic Transformation after Endovascular Thrombectomy in Acute Ischemic Stroke: Phantom and Patient Study.

BACKGROUND AND PURPOSE: Early differentiation of contrast staining from hemorrhagic transformation in patients with acute ischemic stroke who have undergone endovascular treatment is critical in preventing the delayed administration of antiplatelet agents. We aimed to demonstrate the usefulness of an immediate postinterventional DWI protocol including B0 and gradient recalled-echo sequences to discriminate those 2 conditions through phantom and preliminary retrospective patient studies. MATERIALS AND METHODS: On 3T MR imaging, the signal intensities of the phantom models consisting of iodinated contrast agents diluted with normal saline and arterial blood were compared using T1WI, T2WI, and gradient recalled-echo sequences. A total 17 patients (8 with hemorrhagic transformation and 9 with contrast staining; 8 men and 9 women; mean age, 72.00 ± 10.91 years; range, 52-90 years) who underwent mechanical thrombectomy for acute ischemic stroke and showed newly appearing hyperdense lesions on immediate (<24 hours) postinterventional nonenhanced CT scans were included in this study. Immediate postinterventional DWI of patients were compared. RESULTS: In the phantom study, iodinated contrast agents diluted with normal saline showed minimal signal drop, while those diluted with arterial blood demonstrated dark signal intensity in the T2WI and gradient recalled-echo sequences. In the patient study, all hemorrhagic transformations and none of the contrast staining demonstrated dark or low signal (

Evaluation of cardiopulmonary and inflammatory markers in dogs with heartworm infection treated using the slow kill method.

This study evaluated the changes in the levels of cardiac, hemostatic, and inflammatory biomarkers in 12 dogs with different severities of heartworm infection treated using the slow kill protocol, consisting of 6-10µg/kg of ivermectin and 10mg/kg of doxycycline combination. The serum levels of cardiac troponin-I, D-dimer, C-reactive protein, and interleukin-6 were measured on the day of diagnosis (D0), after termination of doxycycline administration (D30), after termination of the slow kill treatment (D180), and 10 months after the initiation of therapy (D300). Heartworm antigenemia was cleared in 4/4 class I dogs, 3/4 class II dogs, and 1/4 class III dogs at the end of the therapy (D180), and in 4/4 class I, 4/4 class II, and 1/4 class III dogs at the end of the study (D300). The serum levels of the markers in class I dogs on the day of diagnosis (D0) were within the reference range, while the levels in class II and III dogs were above the reference range. Further, the serum levels of the markers in all dogs decreased significantly at the end of the study (D300), although some markers in class III dogs remained at pathological levels. This study revealed that the slow kill method should be used only as an alternative therapeutic protocol for dogs with low worm burden (class I and II). As the slow kill method alone may not effectively reduce all pathological changes in dogs with heavy worm burden and severe clinical signs (class III), adjuvant therapies including steroids and anti-thromboembolics should be used to minimize the risk of complications.

Effects of cyclosporine and tacrolimus on the oxidative stress in cultured mesangial cells.

OBJECTIVES: Cyclosporine (CsA) and tacrolimus (Tac) are two primary immunosuppressive agents used for the prevention of graft rejection. However, their use is associated with significant side effects, most notably nephrotoxicity. The mechanisms of this toxicity are not fully understood, but they seem to be associated with increases in the production of oxygen free radicals (OFRs). This present work examined the effect of CsA and Tac on the production of OFRs in cultured rat renal mesangial cells (RMCs). METHODS: Varying concentrations of CsA and Tac (0 to 40 micromol/L) were added to RMCs and incubated for 60 minutes at 37 degrees C. The production of OFRs was evaluated by measuring the fluorescent product from the oxidation of an oxidant-sensitive 2', 7'-dichlorofluorescin. RESULTS: At 60 minutes, the relative fluorescence units (RFU) for OFRs production in RMCs exposure to CsA were increased by 2.5%, 11.5%, 22.5%, 57.2%, and 174% at 2.5, 5, 10, 20, and 40 micromol/L, respectively. Tac increased the RFU by 15.9%, 13.6%, 14.8%, 13.2%, 21.4%, 13.2%, and 28.1% at 0.1, 1, 2.5, 5, 10, 20, and 40 micromol/L, respectively. In RMCs, the RFU produced by CsA was higher than that by Tac. CONCLUSIONS: The results of this experiment suggest that CsA and Tac induced renal injury by OFRs.

Effect of tacrolimus on the production of oxygen free radicals in hepatic mitochondria.

OBJECTIVES: Cyclosporine (CsA) causes side effects that occur mainly in the kidney but also in the liver. Several reports have strongly suggested that the production of oxygen free radicals (OFRs) is a common mechanism of CsA toxicity. However, tacrolimus is believed to suppress the production of OFRs. METHODS: We obtained the mitochondrial fraction with 96% purity from rat liver using a sucrose density gradient solution. Zero to 100 micromol/L tacrolimus was incubated with the mitochondrial fraction for 6 hours at 37 degrees C. OFRs were evaluated by measuring the fluorescent product from the oxidation of an oxidant-sensitive 2,7-dichlorefluorescein using a VICTOR3 multilabel counter. RESULTS: The fluorescence units for OFR production were increased as the time of exposure to tacrolimus passed from 1 to 6 hours. The fluorescence units in 0.1 micromol/L tacrolimus were 6.0 x 10(5) at 1 hour, 7.8 x 10(5) at 2 hours, 9.0 x 10(5) at 3 hours, 10.0 x 10(5) at 4 hours, 11.1 x 10(5) at 5 hours, and 11.4 x 10(5) at 6 hours. However, the fluorescence units were similar although the tacrolimus concentration increases from 0.1 to 100 micromol/L. CONCLUSIONS: The results in this experiment suggested that tacrolimus induced the production of OFRs depending on the exposure time.

p21(WAF1) is associated with CDK2 and CDK4 protein during HL-60 cell differentiation by TPA treatment.

TPA-treated HL-60 cells are mainly arrested in G1 by p21(WAF1) accumulation. We investigate the downstream changes following such accumulation. Increased p21(WAF1) is associated with CDK2 and CDK4. pRb is dephosphorylated in the presence of p21-CDK2/4 complexes, and the Rb-E2F1 complex increases after TPA treatment, whereas the Rb-HDAC1 complex decreases slightly. Our results suggest that increased p21(WAF1) is associated with CDK2/4, and that these complexes induce pRb dephosphorylation. In turn, hypophosphorylated pRb are mainly complexed with E2F1, but HDAC1 appears not to be a key component in this process.

Immune response induced by immunization with Hepatitis B virus core DNA isolated from chronic active hepatitis patients.

There are many mutations in the gene encoding Hepatitis B virus (HBV) core antigen of chronic active hepatitis patients, and such mutations are most likely to be related to the severity of disease. Here, we constructed plasmids containing wild-type and deletion type of HBV core gene (HBc) to develop an experimental DNA vaccine and to compare immunogenicity of two types of HBc vaccine. Twenty-nine wild-types and seven deletion types of HBc were detected in sera of 32 Korean patients with chronic active hepatitis. Four wild-types (W1, W2, W4, W6) and two deletion types (D3, D4) of HBc were cloned into the pcDNA3 vector. Intramuscular immunization with wild-type HBc efficiently increased serum anti-HBc antibody response in a dose-dependent manner. Anti-HBc antibody response in mice injected with W6 increased 14 days after immunization, and peaked after 30 days and was maintained at least up to 50 days. W6 immunization induced a specific cytotoxic T lymphocyte response to W6-transfected 3LL (3LL-W6), and reduced the sizes of tumor mass of mice challenged with 3LL-W6 or 3LL transfected with D4. However, intramuscular immunization with D3 and D4 did not show antibody response at all. D3 and D4 have 157 bp (from 331 to 491 bp) and 122 bp (from 327 to 448 bp) gene deletion, respectively, and these encode class II MHC-restricted T-cell epitope. Altogether, these results suggest that mutant virus that has deleted HBc gene may evade immune systems due to loss of T-cell epitope.

Lack of mutation at p16INK4A gene but expression of aberrant p16INK4A RNA transcripts in human ovarian carcinoma.

Alterations of the p16INK4A gene are frequent in various human cancers. We investigated p16INK4A gene status in 20 ovarian carcinomas by PCR (polymerase chain reaction), PCR-SSCP (polymerase chain reaction-single strand conformation polymorphism) and sequencing techniques. None of the primary tumors showed any mutational or deletional events. However, 19 out of 20 tumors displayed both a methylated and an unmethylated p16INK4A promoter. In some of these samples, we detected aberrant p16INK4A transcripts, with partial deletions of both exons 1 and 2, which could not encode a functional p16INK4A protein. The sequences of the aberrant mRNA revealed common 4-7 nucleotide sequences before and after the deleted region, which might cause abnormal splicing of mRNA transcripts. These results suggest that both promoter methylation and aberrant mRNA processing may interfere with p16INK4A expression in ovarian tumors.

5-Aza-2'-deoxycytidine leads to down-regulation of aberrant p16INK4A RNA transcripts and restores the functional retinoblastoma protein pathway in hepatocellular carcinoma cell lines.

The inactivation of the cyclin-dependent kinase 4 and 6 (CDK4/6) inhibitor p16INK4A may be caused by gene deletion, mutation or promoter hypermethylation. We have previously reported that p16INK4A in hepatocellular carcinoma (HCC) tissues and cell lines is inactivated predominantly by promoter hypermethylation rather than genomic aberrations. In the present experiments, we have studied the effects of the demethylating agent, 5-aza-2'-deoxycytidine (5-AZA/decitabine), on the expression of aberrant p16INK4A RNA transcripts and the CDK-retinoblastoma gene pathway in HCC cell lines with p16INK4A promoter hypermethylation. The expression of aberrant p16INK4A RNA transcripts was down-regulated and p16INK4A protein was strongly re-expressed in the HCC cell lines, SNU 354, 398, 423 and 475 after 5-AZA/decitabine treatment for 5 days. The re-expressed p16INK4A was functional, because it bound to and inhibited CDK4 kinase activity, and increased the concentrations of the hypophosphorylated form of retinoblastoma protein (pRB) in cells with a wild type RB gene. Moreover, treatment with the demethylating agent led not only to G1 cell cycle arrest, but also to the increased expression of the senescence-associated marker beta-galactosidase. This up-regulation of p16INK4A mRNA and protein correlated with demethylation of the p16INK4A promoter, and with the down-regulation or disappearance of aberrant p16INK4A transcripts. These results suggest that the aberrant p16INK4A RNA transcript can be transcribed from the methylated p16INK4A gene, and endogenous reactivation of functional p16INK4A mRNA by a demethylating agent can restore the pRB pathway in HCC, and foster the terminal differentiation of the malignant cells. Therefore, demethylating agents, such as 5-AZA/decitabine, may have potential in the treatment of HCC.

Chemopreventive agent resveratrol, a natural product derived from grapes, reversibly inhibits progression through S and G2 phases of the cell cycle in U937 cells.

Resveratrol, a natural product derived from grapes, has been shown to prevent carcinogenesis in murine models. We report here that resveratrol induces antiproliferation and arrests the S phase in human histiocytic lymphoma U937 cells. Resveratrol induces arrest in the S phase at low concentrations (30-60 microM), but high concentrations do not induce S phase accumulation in U937 cells. Removal of resveratrol from the culture medium stimulates U937 cells to reenter the cell cycle synchronously, as judged by the expression patterns of cyclin E, A and by fluorescent activated cell sorting analysis. These data demonstrate that resveratrol causes S phase arrest and reversible cell cycle arrest. Thus, resveratrol provides an important new cell cycle blocker as well as a cancer chemopreventive agent.

Overexpression of short heterodimer partner recovers impaired glucose-stimulated insulin secretion of pancreatic beta-cells overexpressing UCP2.

The short heterodimer partner (SHP) (NR0B2) is an orphan nuclear receptor whose function in pancreatic beta-cells is unclear. Mitochondrial uncoupling protein (UCP2) in beta-cells is upregulated in obesity-related diabetes, causing impaired glucose-stimulated insulin secretion (GSIS). We investigated whether SHP plays a role in UCP2-induced GSIS impairment. We overexpressed SHP in normal islet cells and in islet cells overexpressing UCP2 by an adenovirus-mediated infection technique. We found that SHP overexpression enhanced GSIS in normal islets, and restored GSIS in UCP2-overexpressing islets. SHP overexpression increased the glucose sensitivity of ATP-sensitive K+ (KATP) channels and enhanced the ATP/ADP ratio. A peroxisome proliferator-activated receptor gamma (PPARgamma) antagonist, GW9662, did not block the SHP effect on GSIS. SHP overexpression also corrected the impaired sensitivity of UCP2-overexpressing beta-cells to methylpyruvate, another energy fuel that bypasses glycolysis and directly enters the Krebs cycle. KATP channel inhibition mediated by dihydroxyacetone, which gives reducing equivalents directly to complex II of the electron transport system, was similar in Ad-Null-, Ad-UCP2- and Ad-UCP2+Ad-SHP-infected cells. The mitochondrial metabolic inhibitor sodium azide totally blocked the effect of SHP overexpression on GSIS. These results suggest that SHP positively regulates GSIS in beta-cells and restores glucose sensitivity in UCP2-overexpressing beta-cells by enhancing mitochondrial glucose metabolism, independent of PPARgamma activation.

The effects of the melatonin on ultraviolet-B irradiated cultured dermal fibroblasts.

It has been reported that reactive oxygen species (ROS) and oxygen-derived free radicals are generated by ultraviolet (UV) radiation and various chemicals and their important roles in cellular damage and apoptosis are being increasingly recognized. Melatonin is a hormone with multiple functions in humans, produced by the pineal gland and stimulated by beta-adrenergic receptors. Melatonin has been shown to have photo protection properties, but there has been little progress toward identifying the specific mechanisms of its action. To clarify the role of melatonin as a free radical scavenger, in response to ultraviolet-B (UVB) irradiation, we investigated the effects of UVB and melatonin on cytotoxicity, lipid peroxidation, terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick end-labeling (TUNEL) assay and alteration of cell cycle in cultured skin fibroblast. Cell survival curves after UVB irradiation showed dose dependent decrement pattern by trypan blue exclusion assay. Only 56% of dermal fibroblasts were survived at 140 mJ/cm2 UVB irradiation. The damage was associated with cell membrane lipid peroxidation, as shown by accumulation malondialdehyde (MDA). By pre-cultivation with melatonin (10(-9) M), a significant preventive effect was noted on the increase in the absolute number of surviving cells (up to 92.5% of cells were survived) and the levels of MDA were markedly decreased. These finding suggest significant correlation between an increase of lipid peroxide and cell viability. Morphological changes associated with apoptotic cell death were easily distinguished by TUNEL stain. Quantitative analysis of DNA content of skin fibroblasts was evaluated by flow cytometric analysis performed after vital staining with propidium iodide. UVB suppresses the G1 progression induced pre-G1 arrest leading to apoptotic changes of dermal fibroblast and those are blocked by melatonin pre-treatment. The results show the photodynamic effects of UVB that supposes the production of ROS and arrest the cell cycle. Melatonin, which have newly accepted as a potential UV protection properties, is effective membrane peroxidation inhibitor and prevent the pre-G1 arrest when present in relevant concentration during UVB irradiation.

The green tea polyphenol (-)-epigallocatechin gallate attenuates beta-amyloid-induced neurotoxicity in cultured hippocampal neurons.

Previous evidence has indicated that the neuronal toxicity of amyloid beta (betaA) protein is mediated through oxygen free radicals and can be attenuated by antioxidants and free radical scavengers. Recent studies have shown that green tea polyphenols reduced free radical-induced lipid peroxidation. The purpose of this study was to investigate whether (-)-epigallocatechin gallate (EGCG) would prevent or reduce the death of cultured hippocampal neuronal cells exposed to betaA because EGCG has a potent antioxidant property as a green tea polyphenol. Following exposure of the hippocampal neuronal cells to betaA for 48 hours, a marked hippocampal neuronal injuries and increases in malondialdehyde (MDA) level and caspase activity were observed. Co-treatment of cells with EGCG to betaA exposure elevated the cell survival and decreased the levels of MDA and caspase activity. Proapoptotic (p53 and Bax), Bcl-XL and cyclooxygenase (COX) proteins have been implicated in betaA-induced neuronal death. However, in this study the protective effects of EGCG seem to be independent of the regulation of p53, Bax, Bcl-XL and COX proteins. Taken together, the results suggest that EGCG has protective effects against betaA-induced neuronal apoptosis through scavenging reactive oxygen species, which may be beneficial for the prevention of Alzheimer's disease.

Effects of polyhemoglobin-antioxidant enzyme complex on ischemia-reperfusion in kidney.

INTRODUCTION: The kidney suffers ischemia-reperfusion (I/R) injury during transplantation. The purpose of the present study was to investigate the therapeutic effect of artificials cells on renal I/R injury through biochemical assays and histological examination. METHODS: We prepared artificial cells using cross-linked hemoglobin (Hb), superoxide dismutase (SOD), and catalase. Normal male Sprague-Dawley rats were divided into 6 groups: the sham-operated control group, the group treated with polyHb,and the group treated with polyHb-SOD-catalase (PSC) (per groups were subjected to ischemia for 1 hour or 2 hours). After reperfusion for 4 hours, kidney and blood samples were obtained. RESULTS: The levels of SOD and catalase in the PSC group were 15 and 50 times higher than those of the control group, respectively. In the polyHb group, the levels of blood urea nitrogen (BUN), serum creatinine, renal hydrogen peroxide, and renal malondialdehyde were increased. However, their levels were significantly decreased by PSC administration. Renal SOD activity did not show any significant changes in the polyHb group, but renal catalase activity was decreased by polyHb treatment in comparison with the control group. The activities of renal SOD and catalase were increased using PSC treatment. In the histological findings, the PSC group showed no evidence of acute tubular necrosis in proximal convoluted tubules; their microvilli and cytoplasmic microorganelles were relatively well preserved. CONCLUSIONS: These results show that PSC effectively reduces renal damage via diminished oxygen free radical-mediated injury after I/R.

Effect of artificial cells on hepatic function after ischemia-reperfusion injury in liver.

BACKGROUND: The liver suffers from ischemia/reperfusion injury during transplantation. Reactive oxygen species generated by xanthine oxidase during reperfusion of the ischemic liver may be partially responsible for the hepatic injury. Oxygen free radicals are removed by antioxidant enzymes such as superoxide dismutase (SOD), catalase, and glutathione peroxidase. Using glutaraldehyde and lysine we constructed crosslinked hemoglobin, containing SOD and catalase, and assessed its ability to protect against ischemia/reperfusion injury during transplantation. METHODS: In contrast to the sham-operated control groups, blood was exchanged using crosslinked hemoglobin (polyHb) a PolyHb-SOD-catalase (PSC) group. After ischemia/reperfusion injury, several parameters of hepatic damage and oxygen free radicals were measured as well as microscopic examination. RESULTS: Alanine aminotransferase, aspartate aminotransferase, superoxide production, hydrogen peroxide, and malondialdehyde levels were higher among the PolyHb group than sham-operated controls. The PolyHb group revealed a few apoptotic bodies, some acute inflammatory infiltrates in the sinusoids, nuclear fragmentations, cell shrinkage, and chromatin clumping with formation of apoptotic bodies in the apoptotic cells under microscopic examination. Alanine aminotransferase, aspartate aminotransferase, superoxide production, and hydrogen peroxide levels were lower in the PSC than the PolyHb group. Hepatic structures were well preserved in the PSC group. CONCLUSIONS: Reactive oxygen species contribute to hepatic dysfunction with morphologic changes. PSC is effective to reduce hepatic damage by lowering oxygen free radical-mediated injury after ischemia/reperfusion in the liver.