PURPOSE: Aspirin(acetylsalicylic acid) has been used to treat unstable angina and acute myocardial infarction in adults and Kawasaki disease in children. The antithrombotic effect of aspirin was attributed to its ability to inhibit platelet aggregation by inhibiting platelet cyclooxygenase, which leads to decreased thromboxane synthesis. The purpose of this study was to evaluate the effect on the platelet aggregation by low dose aspirin in Kawasaki patients and to learn the side effects of low-dose aspirin. METHODS: Fifty patients with Kawasaki disease who were treated with low-dose aspirin, and 22 normal children were studied from Jan. 1996 to Dec. 1997. The platelet count, bleeding time, clotting time, platelet aggregation test(induced by ADP, epinephrine, collagen and ristocetin) and blood aspirin level by colorimetric method were checked. RESULTS: The platelet count, bleeding time, and clotting time in the patient group were not significantly different from the control group. The mean maximum platelet aggregation was 54.4+/-12.8% induced by ADP, 15.9+/-11.7% by epinephrine, 55.5+/-23.8% by collagen, 52.6+/-32.2% by ristocetin in the patient group. It was significantly lower than the control group(P<0.05). The mean blood aspirin level in the patient group was 5.4+/-3.7mg/dl. Side effects of low-dose aspirin were bruise, epistaxis and hematuria. CONCLUSION: Low-dose aspirin therapy in patients with Kawasaki disease inhibited platelet aggregation, but attention would be needed because of the tendency to bleed in these patients. Further investigations should be focused on the subject such as the onset of the maximal antiplatelet effect and time needed for the recovery of platelet function.
Adenosine Diphosphate ; Adult ; Angina, Unstable ; Aspirin* ; Bleeding Time ; Blood Platelets* ; Child ; Collagen ; Contusions ; Epinephrine ; Epistaxis ; Hematuria ; Humans ; Mucocutaneous Lymph Node Syndrome* ; Myocardial Infarction ; Platelet Aggregation* ; Platelet Count ; Prostaglandin-Endoperoxide Synthases ; Ristocetin

Both adaptive and deleterious responses of cells to ethanol are likely triggered by short-term interactions of the cells with ethanol. Many studies have demonstrated the direct effect of ethanol on growth factor-stimulated cell proliferation. Using Swiss 3T3 cells whose growth was inhibited by ethanol in a concentration-dependent manner, we further investigated the molecular mechanisms of acute ethanol treatment by examining its effect on EGF- and PDGF-mediated cellular signaling systems for the mitogenic function. Tyrosine autophosphorylation of the growth factor receptors was partially prevented by ethanol in intact cells. When ethanol was included before or after EGF stimulation, no effect on the receptor signaling was observed. Here we also report that ethanol inhibits activation of ERK induced by both EGF and PDGF. EGF-induced JNK activation was reduced but PDGF-induced rapid JNK activation was delayed by the addition of ethanol. The balance between its inhibitory and stimulatory effect on the signaling molecules might determine the rate of cell growth.
3T3 Cells ; Animal ; Cell Division/drug effects ; Drug Interactions ; Epidermal Growth Factor/pharmacology* ; Ethanol/pharmacology* ; Mice ; Mitogen-Activated Protein Kinases/metabolism ; Mitogens/pharmacology* ; Platelet-Derived Growth Factor/pharmacology* ; Signal Transduction/drug effects

In addition to its well-known glycolytic activity, GAPDH displays multiple functions, such as nuclear RNA export, DNA replication and repair, and apoptotic cell death. This functional diversity depends on its intracellular localization. In this study, we explored the signal transduction pathways involved in the nuclear translocation of GAPDH using confocal laser scanning microscopy of immunostained human diploid fibroblasts (HDFs). GAPDH was present mainly in the cytoplasm when cultured with 10% FBS. Serum depletion by culturing cells in a serum-free medium (SFM) led to a gradual accumulation of GAPDH in the nucleus, and this nuclear accumulation was reversed by the re-addition of serum or growth factors, such as PDGF and lysophosphatidic acid. The nuclear export induced by the re-addition of serum or growth factors was prevented by LY 294002 and SH-5, inhibitors of phosphoinositide 3-kinase (PI3K) and Akt/protein kinase B, respectively, suggesting an involvement of the PI3K signaling pathway in the nuclear export of GAPDH. In addition, 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR), an activator of AMP-activated protein kinase (AMPK), stimulated the nuclear translocation of GAPDH and prevented serum- and growth factor-induced GAPDH export. AMPK inhibition by compound C or AMPK depletion by siRNA treatment partially prevented SFM- and AICAR-induced nuclear translocation of GAPDH. Our data suggest that the nuclear translocation of GAPDH might be regulated by the PI3K signaling pathway acting mainly as a nuclear export signal and the AMPK signaling pathway acting as a nuclear import signal.

Lysophosphatidic acid (LPA) is a phospholipid growth factor that acts through G-protein-coupled receptors. Previously, we demonstrated an altered profile of LPA-dependent cAMP content during the aging process of human diploid fibroblasts (HDFs). In attempts to define the molecular events associated with the age-dependent changes in cAMP profiles, we determined the protein kinase A (PKA) activity, phosphorylation of cAMP-response element binding protein (CREB), and the protein expression of CRE-regulatory genes, c-fos and COX-2 in young and senescent HDFs. We observed in senescent cells, an increase in mRNA levels of the catalytic subunit a of PKA and of the major regulatory subunit Ia. Senescence-associated increase of cAMP after LPA treatment correlated well with increased CREB phosphorylation accompanying activation of PKA in senescent cells. In senescent cells, after LPA treatment, the expression of c-fos and COX-2 decreased initially, followed by an increase. In young HDFs, CREB phosphorylation decreased following LPA treatment, and both c-fos and COX-2 protein levels increased rapidly. CRE-luciferase assay revealed higher basal CRE-dependent gene expression in young HDFs compared to senescent HDFs. However, LPA-dependent slope of luciferase increased more rapidly in senescent cells than in young cells, presumably due to an increase of LPA-induced CREB phosphorylation. CRE-dependent luciferase activation was abrogated in the presence of inhibitors of PKC, MEK1, p38MAPK, and PKA, in both young and senescent HDFs. We conclude that these kinase are coactivators of the expression of CRE-responsive genes in LPA-induced HDFs and that their changed activities during the aging process contribute to the final expression level of CRE-responsive genes.
Time Factors ; Protein Kinase Inhibitors/pharmacology ; Phosphorylation ; Male ; Lysophospholipids/*pharmacology ; Luciferases/genetics/metabolism ; Humans ; Gene Expression/drug effects ; Fibroblasts/cytology/*drug effects/metabolism ; Diploidy ; Cyclic AMP-Dependent Protein Kinases/genetics/metabolism ; Cyclic AMP Response Element-Binding Protein/metabolism ; Cyclic AMP/*metabolism ; Cells, Cultured ; Cell Aging/physiology ; Catalytic Domain/genetics

The treatment of cystamine, a transglutaminase (TGase) inhibitor, has beneficial effects in several diseases including CAG-expansion disorders and cataract. We compared the inhibition characteristics of cystamine with those of cysteamine, a reduced form of cystamine expected to be present inside cells. Cystamine is a more potent inhibitor for TGase than cysteamine with different kinetics pattern in a non- reducing condition. By contrast, under reducing conditions, the inhibitory effect of cystamine was comparable with that of cysteamine. However, cystamine inhibited intracellular TGase activity more strongly than cysteamine despite of cytoplasmic reducing environment, suggesting that cystamine itself inhibits in situ TGase activity by forming mixed disulfides.
Cell Line, Tumor ; Comparative Study ; Cystamine/*pharmacology ; Cysteamine/*pharmacology ; Enzyme Inhibitors/*pharmacology ; Humans ; Research Support, Non-U.S. Gov't ; Transglutaminases/*antagonists & inhibitors

Baicalein is one of the major flavonoids in Scutellaria baicalensis Georgi and possesses various effects, including cytoprotection and anti-inflammation. Because endoplasmic reticulum (ER) stress has been implicated in neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, and cerebral ischemia, we investigated the effects of baicalein on apoptotic death of HT22 mouse hippocampal neuronal cells induced by thapsigargin (TG) and brefeldin A (BFA), two representative ER stress inducers. Apoptosis, reactive oxygen species (ROS) production, and mitochondrial membrane potential (MMP) were measured by flow cytometry. Expression level and phosphorylation status of ER stress-associated proteins and activation and cleavage of apoptosis-associated proteins were analyzed by Western blot. Baicalein reduced TG- and BFA-induced apoptosis of HT22 cells and activation and cleavage of apoptosis-associated proteins, such as caspase-12 and -3 and poly(ADP-ribose) polymerase. Baicalein also reduced the TG- and BFA-induced expression of ER stress-associated proteins, including C/EBP homologous protein (CHOP) and glucose-regulated protein 78, the cleavage of X-box binding protein-1 and activating transcription factor 6alpha, and the phosphorylation of eukaryotic initiation factor-2alpha and mitogen-activated protein kinases, such as p38, JNK, and ERK. Knock-down of CHOP expression by siRNA transfection and specific inhibitors of p38 (SB203580), JNK (SP600125), and ERK (PD98059) as well as anti-oxidant (N-acetylcysteine) reduced TG- or BFA-induced cell death. Baicalein also reduced TG- and BFA-induced ROS accumulation and MMP reduction. Taken together, these results suggest that baicalein could protect HT22 neuronal cells against ER stress-induced apoptosis by reducing CHOP induction as well as ROS accumulation and mitochondrial damage.
Animals ; *Apoptosis ; Brefeldin A/pharmacology ; Cell Line ; Cytoprotection ; DNA-Binding Proteins/metabolism ; Endoplasmic Reticulum/drug effects/*physiology ; Flavanones/*pharmacology ; Heat-Shock Proteins/biosynthesis ; Hippocampus/cytology ; Membrane Potential, Mitochondrial/drug effects ; Mice ; Mitogen-Activated Protein Kinases/metabolism ; Neurons/*drug effects/physiology ; Reactive Oxygen Species/*metabolism ; Signal Transduction/drug effects ; Thapsigargin/pharmacology ; Transcription Factor CHOP/*biosynthesis ; Transcription Factors/metabolism ; Unfolded Protein Response/drug effects

Purpose: In colorectal cancer surgery, it is important to have accurate resection margins. However, it is challenging to localize lesions during laparoscopy. Therefore, to reduce surgical errors, many preoperative localizing methods have been introduced. In this study, we aimed to assess the preoperative feasibility and safety of autologous blood tattooing. Methods: A total of 11 patients underwent preoperative colonoscopic autologous blood tattooing from August 2017 to February 2020. At the start of the surgery, the surgeon assessed the patients for the precision of visibility and other complications such as abscess or spillage. The patients’ characteristics, outcomes, and complications were collected retrospectively. Results: The study comprised 8 men and 3 women, with an average age of 63 years. Ten patients showed precise visibility, and no localization errors were observed during surgery. No complication was observed in all patients. Conclusion Preoperative autologous blood tattooing is a very useful and safe technique because it has high visibility with no complications. This method does not require additional agents or facilities. A large-scale study will be required to develop standard guidelines.