Proangiogenic functions of an RGD-SLAY-containing osteopontin icosamer peptide in HUVECs and in the postischemic brain

Osteopontin (OPN) is a phosphorylated glycoprotein secreted into body fluids by various cell types. OPN contains arginine-glycine-aspartate (RGD) and serine-leucine-alanine-tyrosine (SLAY) motifs that bind to several integrins and mediate a wide range of cellular processes. In the present study, the proangiogenic effects of a 20-amino-acid OPN peptide (OPNpt20) containing RGD and SLAY motifs were examined in human umbilical vein endothelial cells (HUVECs) and in a rat focal cerebral ischemia model. OPNpt20 exerted robust proangiogenic effects in HUVECs by promoting proliferation, migration and tube formation. These effects were significantly reduced in OPNpt20-RAA (RGD->RAA)-treated cells, but only slightly reduced in OPNpt20-SLAA (SLAY->SLAA)-treated cells. Interestingly, a mutant peptide without both motifs failed to induce these proangiogenic processes, indicating that the RGD motif is crucial and that SLAY also has a role. In OPNpt20-treated HUVEC cultures, AKT and ERK signaling pathways were activated, but activation of these pathways and tube formation were suppressed by anti-αvβ3 antibody, indicating that OPNpt20 stimulates angiogenesis via the αvβ3-integrin/AKT and ERK pathways. The proangiogenic function of OPNpt20 was further confirmed in a rat middle cerebral artery occlusion model. Total vessel length and vessel densities were markedly greater in OPNpt20-treated ischemic brains, accompanied by induction of proangiogenic markers. Together, these results demonstrate that the 20-amino-acid OPN peptide containing RGD and SLAY motifs exerts proangiogenic effects, wherein both motifs have important roles, and these effects appear to contribute to the neuroprotective effects of this peptide in the postischemic brain.

Osteopontin Peptide Icosamer Containing RGD and SLAYGLR Motifs Enhances the Motility and Phagocytic Activity of Microglia

Osteopontin (OPN) is a secreted glycoprotein that is expressed in various tissues, including brain, and mediates a wide range of cellular activities. In a previous study, the authors observed the robust neuroprotective effects of recombinant OPN and of RGD and SLAYGLR-containing OPN-peptide icosamer (OPNpt20) in an animal model of transient focal ischemia, and demonstrated anti-inflammatory and pro-angiogenic effects of OPNpt20 in the postischemic brain. In the present study, we investigated the effects of OPNpt20 on the motility and phagocytic activity of BV2 cells (a microglia cell line). F-actin polymerization and cell motility were significantly enhanced in OPNpt20-treated BV2 cells, and numbers of filopodia-like processes increased and lamellipodia-like structures enlarged and thickened. In addition, treatment of cells with either of three mutant OPN icosamers containing mutation within RGD, SLAY, or RGDSLAY showed that the RGD and SLAY motifs of OPNpt20 play critical roles in the enhancement of cell motility, and the interaction between exogenous OPNpt20 and endogenous αv and α4 integrin and the activations of FAK, Erk, and Akt signaling pathways were found to be involved in the OPNpt20-mediated induction of cell motility. Furthermore, phagocytic activity of microglia was also significantly enhanced by OPNpt20 in a RGD and SLAY dependent manner. These results indicate OPNpt20 containing RGD and SLAY motifs triggers microglial motility and phagocytic activity and OPNpt20-integrin mediated signaling plays a critical role in these activities.

Comparative assessment of physicochemical properties of unripe peach (Prunus persica) and Japanese apricot (Prunus mume)

<p><b>OBJECTIVE</b>To investigate the physicochemical properties of unripe peach-Prunus persica cv. Mibaekdo (Mibaekdo) and Prunus persica cv. Nagasawa Hakuho (Nagasawa Hakuho) as an alternative to food supplement while Japanese apricot (Prunus mume cv. Backaha) (Backaha) was used as a control sample.</p><p><b>METHODS</b>The unripe fruits were analyzed for soluble solid ( ˚Brix), titratable acidity, pH, total polyphenol content, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, amygdalin content, free amino acid content, organic acid content, free sugar content, and α-amylase activities.</p><p><b>RESULTS</b>Total polyphenol content of unripe peach ranged between 137.27-151.64 µg/g whereas that of apricot was 160.73 µg/g. DPPH radical scavenging activities of Backaha was the highest (89.16%) followed by Mibaekdo (85.05%) and Nagasawa Hakuho (41.50%). The highest amount of oxalic acid (612.8 mg/100 g) was observed in Mibaekdo while that of Nagasawa Hakuho and Backaha were (184.6±18.1) and (334.8±16.1) mg/100 g, respectively. Amygdalin contents of Mibaekdo, Nagasawa Hakuho and Backaha were 486.61, 548.60 and 174.28 µg/g, respectively.</p><p><b>CONCLUSIONS</b>The results suggest that the unripe fruit of peach has a significant biochemical potential of using as a food supplement with potential health benefit for human health.</p>

Glycyrrhizin Attenuates Kainic Acid-Induced Neuronal Cell Death in the Mouse Hippocampus

Glycyrrhizin (GL), a triterpene that is present in the roots and rhizomes of licorice (Glycyrrhiza glabra), has been reported to have anti-inflammatory and anti-viral effects. Recently, we demonstrated that GL produced the neuroprotective effects with the suppression of microglia activation and proinflammatory cytokine induction in the postischemic brain with middle cerebral artery occlusion (MCAO) in rats and improved motor impairment and neurological deficits. In the present study, we investigated whether GL has a beneficial effect in kainic acid (KA)-induced neuronal death model. Intracerebroventricular (i.c.v.) injection of 0.94 nmole (0.2 microg) of KA produced typical neuronal death in both CA1 and CA3 regions of the hippocampus. In contrast, administration of GL (10 mg/kg, i.p.) 30 min before KA administration significantly suppressed the neuronal death, and this protective effect was more stronger at 50 mg/kg. Moreover, the GL-mediated neuroprotection was accompanied with the suppression of gliosis and induction of proinflammatory markers (COX-2, iNOS, and TNF-alpha). The anti-inflammatory and anti-excitotoxic effects of GL were verified in LPS-treated primary microglial cultures and in NMDA- or KA-treated primary cortical cultures. Together these results suggest that GL confers the neuroprotection through the mechanism of anti-inflammatory and anti-excitotoxic effects in KA-treated brain.

HMGB1-Binding Heptamer Confers Anti-Inflammatory Effects in Primary Microglia Culture

High mobility group box 1 (HMGB1) is an endogenous danger signal molecule. In the postischemic brain, HMGB1 is massively released during NMDA-induced acute damage and triggers inflammatory processes. In a previous study, we demonstrated that intranasally delivered HMGB1 binding heptamer peptide (HBHP; HMSKPVQ) affords robust neuroprotective effects in the ischemic brain after middle cerebral artery occlusion (MCAO, 60 minutes). In the present study, we investigated HBHP-induced anti-inflammatory effects on microglia activation. In LPS-treated primary microglia culture, HMGB1 was rapidly released and accumulated in culture media. Furthermore, LPS-conditioned media collected from primary microglia cultures (LCM) activated naive microglia and markedly induced NO and proinflammatory cytokines. However, the suppression of HMGB1 by siRNA-HMGB1, HMGB1 A box, or anti-HMGB1 antibody significantly attenuated LCM-induced microglial activation, suggesting that HMGB1 plays a critical role in this process. A pull-down assay using biotin-labeled HBHP showed that HBHP binds directly to HMGB1 (more specifically to HMGB1 A box) in LCM. In addition, HBHP consistently inhibited LCM-induced microglial activation and suppressed the inductions of iNOS and proinflammatory cytokines. Together these results suggest that HBHP confers anti-inflammatory effects in activated microglia cultures by forming a complex with HMGB1.

Gene Knockdown in the Olfactory Bulb, Amygdala, and Hypothalamus by Intranasal siRNA Administration / 대한해부학회지

Intranasal administration provides a method of bypassing the blood brain barrier, which separates the systemic circulating system and central interstitial fluid, and directly delivering drugs to the central nervous system. This method also circumvents first-pass elimination by the liver and gastrointestinal tract. In the present study, the authors investigated intranasal siRNA delivery efficiency by using FITC-labeled transfection control siRNA and a genespecific siRNA. The localization of fluorescence-tagged siRNA revealed that siRNA was delivered to cells in the olfactory bulb and that the level of the siRNA target gene (alpha B-crystallin) was significantly reduced in the same area. siRNA was delivered to processes as well as nuclei and cytoplasm. At 12 hrs after intranasal delivery, siRNA-mediated target gene reduction was observed in other more distally located brain regions, for example, in the amygdala, entorhinal cortex, and hypothalamus. Target gene knockdown was demonstrated by double immunohistochemistry, which demonstrated alpha B crystallin expression depletion in more than 70% of cells at 12 hrs after the intranasal delivery. siRNA-mediated target gene suppression was detected not only in neurons but in glia, for example, astrocytes. These results indicate that intranasal siRNA delivery offers an efficient means of reducing specific target genes in certain regions of the brain and of performing gene knockdown-mediated therapy.

Risk Factors for Post-ERCP Pancreatitis in Patients Pretreated with Nafamostat Mesilate / 대한소화기내시경학회지

BACKGOUND/AIMS: Pancreatitis is the most common and important complication of an endoscopic retrograde cholangiopancreatography (ERCP). The aim of this study was to identify risk factors for post ERCP-pancreatitis in patients pretreated with nafamostat mesilate, a synthetic protease inhibitor. METHODS: A total of 247 patients who underwent an ERCP were evaluated prospectively. Potential risk factors of post-ERCP pancreatitis in patients pretreated with nafamostat mesilate were evaluated. RESULTS: Twenty-four patients (9.7%) and nine patients (3.6%) developed post-ERCP hyperamylasemia and pancreatitis, respectively. As determined by univariate analysis among the potential risk factors, we found a procedure time over 20 minutes, pancreatic duct cannulation over four times, prior post-ERCP pancreatitis and the absence of a common bile duct (CBD) stone as risk factors for post-ERCP hyperamylasemia. We also found a patient age under 60 years, a procedure time over 20 minutes, pancreatic duct cannulation over four times and the absence of a CBD stone as risk factors for post-ERCP pancreatitis (p<0.05). As determined by multivariate analysis, pancreatic cannulation over four times is independently associated with post-ERCP hyperamylasemia (p=0.038; OR, 5.165; 95% CI, 1.093~24.412) and post-ERCP pancreatitis (p=0.002; OR, 33.122; 95% CI, 3.526~311.138). CONCLUSIONS: A repeated pancreatic duct cannulation is the most important risk factor for post-ERCP pancreatitis in patients pretreated with nafamostat mesilate.

A Case of Hemophagocytic Lymphohistiocytosis Accompanied by Acute Hepatitis A: Review of the Literature / 대한혈액학회지

A previously healthy 26-year-old female was referred to our hospital because of fever and abnormalities of her blood biochemistry. Her laboratory results displayed leukopenia (1.79x103/microliter), thrombocytopenia (85x103/microliter), the serum aspartate aminotransferase and alanine aminotransferase levels were elevated to more than 2,000 IU/L, and the serum levels of lactate dehydrogenase and ferritin were markedly increased. Mild hepatosplenomegaly was reported on the abdomen-pelvis computed tomography. The bone marrow smears revealed proliferation of mature histiocytes that were ingesting platelets and erythrocytes, which is consistent with hemophagocytic lymphohistiocytosis. Although the other viral markers were all negative, the anti-hepatitis A IgM was positive and the anti-hepatitis A IgG was negative. Therefore, the patient was diagnosed as hepatitis A-associated hemophagocytic lymphohistiocytosis. Since a fulminant clinical course was suspected, 2 cycles of cyclosporine (3mg/kg iv from day 1 to day 5), dexamethasone (30mg iv qd from day1 to day 4) and immunoglobulin (500mg/kg iv day 1) therapy was started from the seventh day after onset and a favorable clinical outcome resulted.

SP1 Mediates IFNgamma-induced TIMP-1 Transctription in C6 Astroglioma Cells / 대한해부학회지

Tissue inhibitors of metalloproteinases (TIMPs) comprise a family of secreted multifunctional proteins that consists of four members (TIMP-1 to TIMP-4). TIMPs are all major inhibitors of most matrix metalloproteinases (MMPs). They are synthesized by a variety of different cells and regulated by a number of cytokines and by growth and differentiation factors. The balance between MMPs and TIMPs plays a crucial role in the turnover of extracellular matrix in normal and pathological conditions. Here, we report that the production of TIMP-1 was upregulated in interferon (IFNgamma-treated C6 astroglioma cells and that the proximal 226 bp region of the promoter of the TIMP-1 gene is responsible for IFNgamma-induced induction in C6 astroglioma cells. The induction of TIMP-1 production by IFNgamma was virtually abolished by introducing mutations into the putative SP1-response element in the promoter, indicating that the SP1 binding site conferred responsiveness onto a heterologous promoter. Together the results suggest that the IFNgamma-induced upregulation of TIMP-1 production in C6 astroglioma cells is mediated by the SP1 binding site localized in the TIMP-1 gene promoter.

A Case of Partial Agenesis of Dorsal Panacreas / 대한소화기내시경학회지

Complete or partial agenesis of dorsal pancreas has been reported in a small number of pediatric and adult patients. A case of partial agenesis of dorsal pancreas was reported. This case was not associated with diabetes mellitus and pancreatic exocrine dysfunction, or abdominal pain. A 37-year-old man with chronic hepatitis B visited to our hospital due to further examination for chronic hepatitis B. Abdominal ultrasonography showed enlarged pancreatic head. Diagnosis was made by endoscopic retrograde cholangiopancreatography and computed tomography. Explor-laparotomy was not done.

Two Cases of Ectopic Gallstones Treated by Endoscopic Therapy / 대한소화기내시경학회지

Two cases are herein reported involving patients with ectopic gallstones which were discharged into the stomach and duodenum through a cholecystoduodenal fistula and successfully removed by endoscopic therapy. In the first case, a 75-year-old man was admitted with epigastric pain. Simple abdomen film demonstrated a round laminated calcification and air biliarygram in the RUQ. Endoscopic examination revealed a fistula on the posterior wall of the duodenal bulb and a brown stone (about 5 cm in diameter) was found in the second portion of the duodenum, It was demolished through endoscopic electrohydraulic lithotripsy (EEH1) and discharged with the stool. In the second case, a 55-year-old man was admitted with epigastric pain. A CT scan revealed an ovoid laminated calcification in the dependent portion of the stomach. Endoscopic examination revealed a fistula on the anterior wa11 of' the duodenal bulb and a black pigmented stone (about 2.5 cm in diameter) was found in the stomach. This stone was removed orally by an endoscopic snare. These patients were discharged and remained asymptomatic.

A Case Rhabdomyolysis by Rare Causes: After Consumption of Common Doses of Acetaminophen and Sauna / 대한신장학회잡지

A 21-year-young man had an episode of myalgia and chilling 3 days prior to hospital admission. He had consumed common doses of acetaminophen for one day, and was presented in the sauna room for an hour. On the next morning, he complained of dyspnea and was admitted. He presented in rhabdo-myolysis and acute renal failure with increased up-take in the proximal muscles by (99m)Tc-MPD bone scan. He was treated by hemodialysis and discharged on the twenty-eighth hospital day. Rhabdomyolysis has the variable causes. The causes of this case are two, the first cause is common doses of acetaminophen. But, there is no reports for rhabdomyolysis by common doses of acetaminophen only. However, we should consider that acetaminophen is a contributing factor in this case. The second cause is viral infection. Our patient had myalgia and chilling prior to hospital admission. Heat- stroke is well known cause of rhabdomyolysis. The mechnisms for rhabdomyolysis in this disease are hypovolemia, total body potassium deficit, and increased variable cytokines. Sauna, the last cause of our rhabdomyolysis case may have the same mechanisms with heatstroke. Our case had two causative factors, common doses of acetaminophen and sauna. These factors might be cooperated in our case of rhabdomyolysis and acute renal failure.