BACKGROUND: Kojic acid was used for decades in the cosmetic industry as an antimelanogenic agent. However, there are two major drawbacks of Kojic acid, one is cytotoxicity and second are instability on storage. These limitations led the scientist to synthesize the active Kojic acid peptides. OBJECTIVE: In the present study, we synthesize and investigate the effect of five Kojic acid peptides to overcome the limitation of Kojic acid. METHODS: The peptide was analyzed and purified by high-performance liquid chromatography and matrix-assisted laser desorption ionization time of flight mass spectroscopy. Further, the tyrosinase activities of the Kojic acid and Kojic acid peptides were compared. The toxicity was measured and the melanin content is recorded in B16F10 mouse melanoma cells. RESULTS: Maximum tyrosinase activity was measured by Kojic acid peptides. Therefore, Kojic acid peptides were subjected to melanin assay and cytotoxicity assay and finally the stability of the Kojic acid peptide was measured. CONCLUSION: It was observed that this newly synthesized Kojic acid peptide is stable and potent to inhibit the tyrosinase activity and melanin content of B16F10 mouse melanoma cells without exhibiting cell toxicity. Together, these preliminary results suggest that a further exploration is being needed to establish Kojic acid peptide as antimelanogenic agent.
Animals ; Chromatography, Liquid ; Mass Spectrometry ; Melanins ; Melanoma ; Mice ; Monophenol Monooxygenase ; Peptides

OBJECTIVE: To evaluate their roles in the pathophysiology of preterm premature rupture of membranes (PPROM), we checked interleukin (IL)-6, lipid peroxide, oxygen-radical absorbance capacity (ORAC) and antioxidant vitamin in the venous plasma and amniotic fluid of women with PPROM. METHODS: Venous plasma and amniotic fluid was taken from 20 normal pregnant women and 20 PPROM pregnancy women. IL-6 levels was determined by enzyme-linked immunosorbent assay. Lipid peroxide levels were measured by thiobarbituric acid reaction. The ORAC levels and the antioxidant levels were measured by Cao's method and by high performance liquid chromatography. RESULTS: The IL-6 levels in the venous plasma and amniotic fluid of PPROM pregnancy women were significantly higher than normal pregancy (P<0.01, P<0.01). The lipid peroxide levels in the venous plasma and amniotic fluid of PPROM pregnancy women were significantly higher than normal pregancy (P<0.01, P<0.01). The ORAC values in the venous plasma and amniotic fluid of PPROM pregnancy women were significantly higher than normal pregancy (P<0.01, P<0.01). The ORAC values/lipid peroxide levels in the venous plasma and amniotic fluid of PPROM pregnancy women were significantly higher than normal pregancy (P<0.01, P<0.01). The ascorbic acid levels in the venous plasma and amniotic fluid of PPROM pregnancy women were significantly higher than normal pregancy (P<0.05, P<0.01). CONCLUSION: This result suggest that the increased inflammatory bioactivity, increased lipid peroxidation and decreased antioxidant activity may be involved in the pathophysiology of PPROM. Low levels of ascorbic acid appears to be an important determinant of PPROM.
Amniotic Fluid ; Ascorbic Acid ; Chromatography, Liquid ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Interleukin-6 ; Interleukins ; Lipid Peroxidation ; Membranes ; Plasma ; Pregnancy ; Pregnant Women ; Rupture ; Thiobarbiturates ; Vitamins

OBJECTIVE: To investigate the lipid peroxide levels and protein carbonyls levels in the amniotic fluid of pregnant women with preterm premature rupture of membranes (PPROM). METHODS: The lipid peroxide levels in the amniotic fluid of normal pregnancy (n=20) and pregnant women with PPROM (n=20) were measured by thiobarbituric acid reaction. The protein carbonyl contents in the amniotic fluid of normal pregnancy (n=20) and pregnant women with PPROM (n=20) were determined by the 2,4-dinitrophenylhydrazine method. After amniotic fluid of them were mixed and incubated up to 5 hours with 0.2 mL of 1mM moxalactam, cefodizime, amoxacillin, erythromycin, the lipid peroxide levels and protein carbonyl contents in them were measured. RESULTS: 1. The lipid peroxide levels in the amniotic fluid of pregnant women with PPROM was significantly higher than that of normal pregnancy (9.74+/-0.48 vs. 7.20+/-0.38 nmol/mg protein, P<0.01). 2. The protein carbonyl levels in the amniotic fluid of pregnant women with PPROM was significantly higher than that of normal pregnancy (13.0+/-0.33 vs. 11.27+/-0.17 nmol/mg protein P<0.01). 3. The lipid peroxide levels and protein carbonyls formation by moxalactam in the amniotic fluid of pregnant women with PPROM was significantly higher than basal level (12.08+/-0.81 vs. 9.74+/-0.48 nmol/mg protein, 20.08+/-0.66 vs. 13.0+/-0.33 nmol/mg protein, P<0.01). 4. The lipid peroxide levels and protein carbonyls formation by cefodizime in the amniotic fluid of pregnant women with PPROM was significantly lower than basal level (5.04+/-0.33 vs. 9.74+/-0.48 nmol/mg protein, 9.76+/-0.35 vs. 13.0+/-0.33 nmol/mg protein, P<0.01). 5. There were no significant differences in the levels of lipid peroxide and protein carbonyls by amoxacillin and erythromycin in the amniotic fluid of pregnant women with PPROM between antibiotics-induced and basal levels. CONCLUSION: The lipid peroxidation and the protein carbonyls formation were increased in the amniotic fluid of pregnant women with PPROM. Antibiotics-induced lipid peroxide and protein carbonyl levels were changed in the amniotic fluid of pregnant women with PPROM. Further studies on our results may be beneficial in the selection of antibiotics for pregnant women with PPROM.
Amniotic Fluid ; Anti-Bacterial Agents ; Cefotaxime ; Erythromycin ; Female ; Humans ; Lipid Peroxidation ; Membranes ; Moxalactam ; Phenylhydrazines ; Pregnancy ; Pregnant Women ; Protein Carbonylation ; Rupture ; Thiobarbiturates

Lysophosphatidic acid (LPA) is a bioactive phospholipids and involves in various cellular events, including tumor cell migration. In the present study, we investigated LPA receptor and its transactivation to EGFR for cyclooxygenase-2 (COX-2) expression and cell migration in CAOV-3 ovarian cancer cells. LPA induced COX-2 expression in a dose-dependent manner, and pretreatment of the cells with pharmacological inhibitors of Gi (pertussis toxin), Src (PP2), EGF receptor (EGFR) (AG1478), ERK (PD98059) significantly inhibited LPA- induced COX-2 expression. Consistent to these results, transfection of the cells with selective Src siRNA attenuated COX-2 expression by LPA. LPA stimulated CAOV-3 cell migration that was abrogated by pharmacological inhibitors and antibody of EP2. Higher expression of LPA2 mRNA was observed in CAOV-3 cells, and transfection of the cells with a selective LPA2 siRNA significantly inhibited LPA-induced activation of EGFR and ERK, as well as COX-2 expression. Importantly, LPA2 siRNA also blocked LPA-induced ovarian cancer cell migration. Collectively, our results clearly show the significance of LPA2 and Gi/Src pathway for LPA-induced COX-2 expression and cell migration that could be a promising drug target for ovarian cancer cell metastasis.
Butadienes/pharmacology ; Cell Line, Tumor ; Cell Movement/drug effects/*physiology ; Cyclooxygenase 2/*biosynthesis ; Extracellular Signal-Regulated MAP Kinases/antagonists & inhibitors/metabolism ; Female ; Flavonoids/pharmacology ; GTP-Binding Protein alpha Subunits, Gi-Go/antagonists & inhibitors/*metabolism ; Humans ; Lysophospholipids/pharmacology ; Nitriles/pharmacology ; Ovarian Neoplasms/metabolism/*pathology ; Pertussis Toxin/pharmacology ; Protein-Tyrosine Kinases/antagonists & inhibitors/*metabolism ; Proto-Oncogene Proteins/antagonists & inhibitors/*metabolism ; Pyrimidines/pharmacology ; Receptor, Epidermal Growth Factor/antagonists & inhibitors/metabolism ; Receptors, Lysophosphatidic Acid/*metabolism ; Receptors, Prostaglandin E/metabolism ; Signal Transduction ; Transcriptional Activation ; Tyrphostins/pharmacology

PURPOSE: This study evaluated the influence of bone marrow cell collection techniques and donor site locations on the in-vitro growth of bone-forming cells. METHODS: Sixty six samples of bone marrow cell collections (BMCC) or bone marrow aspirates (BMA) from 15 patients were obtained. Thirty eight samples for culture were composed of 23 BMA from 7 tibial condyles and 16 ilia, with the other 15 BMCC from the contralateral ilia. The other 28 samples were used for the analysis of alkaline phosphatase activities. After counting total cell number, mesenchymal stem cells (MSC) obtained from samples were incubated for 14 days. Alkaline phosphatase staining was used to count the number of stained colonies to show osteogenic differentiation. RESULTS: The average MSC counts of BMA from tibial condyles and ilia were 1.42x10(6) and 7.35x10(6) respectively, with 4.80x10(6) from ilial BMCC (p=0.010). MSC cultures could not be produced from tibial condyles in all 7 samples. However, 9 of 15 BMCC samples and 9 of 16 ilial BMA samples were successfully cultured (p=0.018). The average of cell counts in the successful cultures was 7.92x10(6), whereas that in the failed cultures was 2.85x10(6) (p=0.000). Multiple regression analysis showed that colony count was associated with the patient's age and total cell numbers, but not with collection methods such as BMCC or BMA (p=0.000, R=0.648, beta; age=-0.405, cell number=0.356). The discriminating formula indicated that more than 5.25x10(6) cells were needed for successful culture. CONCLUSIONS: For successful cultures in vitro and for grafts, the total number of collected bone forming cells is more important than donor sites or collection methods. For young patients, grafting of bone-marrow-derived osteoprogenitor cells is promising.
Alkaline Phosphatase ; Bone Marrow ; Bone Marrow Cells ; Cell Count ; Durapatite ; Humans ; Mesenchymal Stromal Cells ; Tissue Donors ; Transplants

OBJECTIVE: Our purpose was to investigate Interleukin-6 (IL-6), tumor necrotic factor-alpha (TNF-alpha), lipid peroxide levels, oxygen-radical absorbance capacity (ORAC), and antioxidant levels in umbilical venous blood plasma and to evaluate the roles of them in the pathophysiology of preeclampsia. STUDY DESIGN: Samples of umbilical venous plasma were obtained from 20 normal and 20 preeclamptic women between 33 and 40 weeks gestation. IL-6 and TNF-alpha was assayed by an enzyme-linked immunoassay. Lipid peroxide levels were measured by thiobarbituric acid reaction. The ORAC values were measured by Cao's method. Ascorbic acid, retinol, alpha-tocopherol, and gamma-tocopherol were measured by high performance liquid chromatography. RESULTS: There was no significant differences of IL-6 levels in umbilical venous plasma between women with normal and preeclampsia (2.79+/-0.21 vs. 2.94+/-0.17 ng/ml). TNF-alpha levels in umbilical venous plasma of women with preeclampsia were significantly higher than that of women with preeclampsia (3.04+/-0.01 vs. 1.40+/-0.01 ng/ml, p<0.01). Lipid peroxide levels in umbilical venous plasma of women with preeclampsia were significantly higher than that of women with normal pregnancy (7.32+/-0.09 vs. 5.18+/-0.14, p<0.01). The ORAC values in umbilical venous plasma of women with preeclampsia were significantly lower than that of women with normal pregnancy (12,836.5+/-249.4 vs. 10,490.2+/-276.9 U/ml, p<0.05). Ascorbic acid levels in umbilical venous plasma of women with preeclampsia were significantly lower than those of women with normal pregnancy (320.2+/-48.5 vs. 538.5+/-68.2 nmol/ml, p<0.05). CONCLUSION: The above results in umbilical venous plasma suggest that the imbalance of lipid peroxidation and antioxidant activity in placenta is involved in the pathophysiology of preeclampsia. Increased TNF-alpha in the umbilical venous plasma showed inflammatory reaction in the placenta would be one of the cause of preclampsia. An antioxidant vitamin, ascorbic acid, may act an important antioxidant factor in preeclampsia.
alpha-Tocopherol ; Ascorbic Acid ; Chromatography, Liquid ; Female ; gamma-Tocopherol ; Humans ; Immunoassay ; Interleukin-6* ; Lipid Peroxidation ; Placenta ; Plasma* ; Pre-Eclampsia* ; Pregnancy ; Tumor Necrosis Factor-alpha* ; Vitamin A ; Vitamins

BACKGROUND: Essential thrombocythemia (ET) is thought to reflect transformation of a multipotent hematopoietic stem cell, but its molecular pathogenesis remains obscure. But tyrosine kinase, especially Janus kinase 2 (JAK2), has been implicated in myeloproliferative disorders other than chronic myeloid leukemia. We investigated the frequency of JAK2 mutation and its correlation with other clinicopathologic variables in Korean patients with ET and reactive thrombocytosis (RT). METHODS: JAK2 mutation analysis was performed on genomic DNA from bone marrow aspirates of 24 patients with ET and peripheral blood in 36 patients with RT using allele-specific PCR. RESULTS: JAK2 mutation was detected in 11 patients (46%) among the 24 patients with ET and was not found in 36 patients with RT. In patients with ET, older age and leukocytosis were related with JAK2 mutation without statistical significance (P=0.172 and 0.094, respectively). But this mutation was not correlated with sex, hemoglobin, platelet count, splenomegaly, increased cellularity of bone marrow, bone marrow fibrosis and vascular complications. CONCLUSIONS: The current observation strengthens the specific association between JAK2 mutation and ET. At the diagnosis of ET in Korean patients, identification of JAK2 mutation should be incorporated in the basis for new approaches.
Aged ; Alleles ; Amino Acid Substitution ; Blood Platelets/metabolism ; Female ; Humans ; Janus Kinase 2/*genetics ; Korea ; Male ; Middle Aged ; Polymerase Chain Reaction ; Thrombocytosis/blood/*diagnosis/genetics

BACKGROUND: We investigated the Janus kinase 2 (JAK2) mutation and its diagnostic value in patients suffering with non BCR/ABL myeloproliferative diseases (nMPD) or other reactive conditions. METHODS: We reviewed the clinical records of 83 patients who underwent bone marrow (BM) examinations with suspect of nMPD. The diagnoses of nMPD were made based on the WHO criteria since 2001 and the PVSG criteria before 2001. The JAK2 mutation was examined by PCR in 54 patients whose BM samples were available. RESULTS: The JAK2 mutation was detected in 25 patients (46%); 12 of 26 patients with essential thrombocythemia (ET), 9 of 12 patients with polycyhtemia vera (PV), one of 7 patients with chronic idiopathic myelofibrosis (CIM) and one patient with unclassifiable MPD. Additionally, JAK2 mutation was detected in each one patient with secondary polycythemia and reactive thrombocytosis. These two patients and two other patients among the JAK2 mutated ET did not meet the WHO PV criteria due to their initial low hemoglobin levels. These patients had liver cirrhosis and hypersplenism due to Budd-Chiari syndrome (1), gastrointestinal bleeding (1) or the initial hemoglobin level was slightly below the level as provided by the criteria, but the level showed a rising pattern despite cytoreductive therapy (2). With the results of the JAK2 mutation available, 4 patients' disease could be re-diagnosed as PV. Finally, the positive rate of the JAK2 mutation was 81% in PV, 48% in ET and 14% in CIM. The presence of JAK2 mutation closely correlated with PV (p=0.001), leukocytosis (p=0.001) and an increased cellularity of BM (p=0.024). CONCLUSIONS: The JAK2 mutation may help differentiate nMPD from secondary cytosis. Therefore, it should be incorporated into the guidelines for the nMPD work-up for making a more accurate diagnosis and administering proper treatment.
Retrospective Studies ; Proto-Oncogene Proteins c-bcr ; Polymerase Chain Reaction ; Myeloproliferative Disorders/*diagnosis/genetics/metabolism ; *Mutation ; Middle Aged ; Male ; Janus Kinase 2/*genetics ; Humans ; Genes, abl ; Female ; Diagnosis, Differential ; DNA/*genetics ; Biological Markers/metabolism ; Aged, 80 and over ; Aged ; Adult

OBJECTIVE: This study was performed to compare the prooxidative activity stimulating the protein carbonyl formation by 3rd generation cephalosporin (moxalactam) and amoxacillin in the uterine venous, umbilical venous, and umbilical arterial plasma of preeclampsia with that of normal pregnancy. METHODS: Lipid peroxide levels in the uterine venous, umbilical venous, and umbilical arterial plasma of normal pregnancy (n=16) and preeclampsia (n=16) were measured by thiobarbituric acid reaction. The basal protein carbonyl contents in the uterine venous, umbilical venous, and umbilical arterial plasma of normal pregnancy (n=16) and preeclampsia (n=16) were determined by the 2,4-dinitrophenylhydrazine (DNPH) method. After plasma of them were mixed and incubated up to 5 hours with 0.2 mL of 1 mM moxalactam or amoxacillin, the protein carbonyl contents in them were measured by DNPH. RESULTS: Lipid peroxide levels in the uterine venous plasma, umbilical venous plasma, and umbilical arterial plasma of preeclampsia were significantly higher than those of normal pregnancy (3.11+/-1.21 vs. 2.18+/-1.16 nmol/mg protein, p<0.05, 5.85+/-1.67 vs. 3.79+/-1.66 nmol/ mg protein, p<0.01, 6.00+/-1.91 vs. 4.99+/-1.78 nmol/mg protein, p<0.01). Protein carbonyls formation by moxalactam in the uterine venous plasma, umbilical venous plasma, and umbilical arterial plasma of preeclampsia were signigicant higher than those of normal pregnancy (19.69+/-8.43 vs. 10.84+/-3.00 nmol/mg protein, p<0.01, 18.94+/-6.96 vs. 10.63+/-1.81 nmol/mg protein, p<0.01, 14.62+/-5.77 vs. 11.21+/-2.08 nmol/mg protein, p<0.05). There were significant positive correlations between lipid peroxide and moxalactam-induced protein carbonyls levels of the uterine venous plasma, umbilical venous plasma, and umbilical arterial plasma (p<0.01). CONCLUSION: These results suggest that increase in the prooxidative activity stimulating the oxidative modification of proteins in utero-placental unit may be involved in the pathogenesis of preeclampsia.
Moxalactam ; Plasma* ; Pre-Eclampsia* ; Pregnancy ; Protein Carbonylation

BACKGROUND: The previous studies suggested that the rate of the people who reported having sensitivity to cold and Raynaud's disease were 12% and 5~10% of general population respectively. There are surprisingly few data on 'cold hands and feet syndrome', the conventional circulatory disorder in far-eastern Asia in spite of the assumption that it is a common disorder among Korean population. Besides, the prevalence of Raynaud's disease has not been studied for Korean complaint of the syndrome. Therefore, we conducted the survey regarding the frequency of Baynaud's disease and phenomenon among the people appealed abnormal sensory symptoms on hands and feet, and 'cold hands and feet syndrome'. METHODS: The questionnaires were asked to 1,008 out- patients who complained abnormal sensory symptoms on hands and feet. The subjects were screened among the patients who visited the clinics, mainly department of family medicine in Uijongbu St. Mary's hospital from January to July 2003. RESULTS: The most common abnormal sensory symptoms were tingling feeling and coldness, pain in the descending order. The mean age of the patients is 47 years, the average age at the onset of the symptoms is 40 years, and the mean duration they had been suffered by the symptoms is 7 years. During interview, 289 patients voluntarily appealed 'cold hands and feet syndrome' in expression of "I have cold extremities" or "I have the syndrome". Among the screened patients, 510 patients answered, "yes" to the question "Are your fingers unusually sensitive to cold?" Two hundred and nineteen patients (43%) of them showed Raynaud's phenomenon. One hundred and sixty patients were diagnosed as Raynaud's disease among 219 patients who had Raynaud's phenomenon. The rest 59 patients were diagnosed as secondary Raynaud's phenomenon. Eighty-seven patients (30%) of 289 patients who voluntarily reported 'cold hands and feet syndrome' were diagnosed as Raynaud's disease. The average age of Raynaud's disease patients is 37 years old, average first attack age is 24 years, and average suffering period is 14 years. Only 10% of cases experienced the first attacks after 40 years old. The attack rates are different by gender. The ratio is 2.3 and higher in female. Only six patients (4%) of Raynaud's disease had been diagnosed accurately. Fifty-three patients (33%) have family history and 41 of them are the first-degree relatives. The causes of secondary Raynaud's phenomenon (59 patients) are connective tissue disorders, neurologic diseases, carpal tunnel syndrome, hypothyroidism, vascular disorders, etc. The statistically significant factors that influence on Raynaud's disease are female, below 40 years old, stress, family history, hypertension, diabetes, etc. The statistical analysis in this study does not prove the association between the disease and the factors such as physical labor, manual labor using hands heavily, smoking, drinking, thyroid diseases, joint disorders, ischemic heart disease, depression. CONCLUSION: Raynaud's disease has been underlooked even though it is not uncommon and can be easily diagnosed and treated. In conclusion, the result of our study suggests that abnormal sensory symptoms on hands and feet, especially 'cold hands and feet syndrome', the conventional circulatory disorder, are the medical condition where special concern on Raynaud's disease is required.
Adult ; Asia ; Carpal Tunnel Syndrome ; Connective Tissue ; Depression ; Diagnosis* ; Drinking ; Female ; Fingers ; Foot* ; Hand* ; Humans ; Hypertension ; Hypothyroidism ; Joints ; Myocardial Ischemia ; Nervous System Diseases ; Prevalence ; Raynaud Disease* ; Sensation* ; Smoke ; Smoking ; Thyroid Diseases ; Surveys and Questionnaires