In this study, the trypsin gene (bgtryp-1) from the German cockroach, Blattella germanica, was cloned via the immunoscreening of patients with allergies to cockroaches. Nucleotide sequence analysis predicted an 863 bp open reading frame which encodes for 257 amino acids. The deduced amino acid sequence exhibited 42-57% homology with the serine protease from dust mites, and consisted of a conserved catalytic domain (GDSGGPLV). bgtryp-1 was determined by both Northern and Southern analysis to be a 0.9 kb, single-copy gene. SDS-PAGE and Western blotting analyses of the recombinant protein (Bgtryp-1) over-expressed in Escherichia coli revealed that the molecular mass of the expressed protein was 35 kDa, and the expressed protein was capable of reacting with the sera of cockroach allergy patients. We also discussed the possibility that trypsin excreted by the digestive system of the German cockroach not only functions as an allergen, but also may perform a vital role in the activation of PAR-2.
Allergens/analysis/chemistry/*genetics ; Amino Acid Sequence ; Base Sequence ; Blattellidae/*genetics ; Blotting, Western ; Catalytic Domain/genetics ; DNA, Complementary/genetics ; Escherichia coli/metabolism ; Female ; *Genes, Insect ; Humans ; Male ; Molecular Sequence Data ; Molecular Weight ; Receptor, PAR-2/metabolism ; Recombinant Proteins/analysis/biosynthesis/chemistry ; Research Support, Non-U.S. Gov't ; Sequence Alignment ; Sequence Homology, Amino Acid ; Trypsin/analysis/chemistry/*genetics

OBJECTIVES: Recently, recombinant FSH (rFSH) has been manufactured using a Chinese hamster ovary cell line transfected with the gene encoding human FSH. Both rFSH and urinary gonadotropin (uFSH) could be used for controlled ovarian hyperstimulation (COH). However, uFSH implies a number of disadvantages, such as batch-to-batch inconsistency, no absolute source control, dependence on large amounts of urine, low specific activity, and low purity. The purpose of this study was to evaluate the efficacy of rFSH in human IVF-ET program. MATERIALS AND METHODS: A total of 508 infertile women was enrolled in this study. They are classified into rFSH group (n=177) or uFSH group (n=331), and all of them were matched by age and cause of infertility in same period. The Puregon(R) (Organon, Holland) was used as rFSH, and the Metrodin-HP(R) (Serono, Switzeland) and Humegon(R) (Organon, Holland) was used as uFSH. We subdivided the patients into three age groups. The outcomes of IVF-ET program were analyzed using the statistical package for social sciences (SPSS). RESULTS: There was no significant differences in the level of estradiol on hCG injection day, the numbers of retrieved oocytes, matured oocytes, fertilized oocytes, transferred embryos, frozen embryos between the two groups. The total dose (IU) of gonadotropin for COH was significantly lower in the rFSH group compared to uFSH group (1339+/-5491.1 vs 2527.8+/-1075.2 IU, p<0.001). Clinical pregnancy rate per embryo transfer in the rFSH group showed increasing tendency, compared to the uFSH group, but there was no statistical significance (35.2% vs 29.3%). Our results demonstrated that the relative efficiency of rFSH compared with uFSH is higher in older patients. CONCLUSIONS: The ovarian stimulatory effect and clinical outcome of recombinant FSH was similar to that of the urinary gonadotropin. The IVF-ET cycles with significantly lower dose of gonadotropin in rFSH group showed comparable results. Therefore, we suggest that recombinant FSH is more potent and effective than urinary gonadotropin.
Animals ; Cell Line ; Cricetinae ; Cricetulus ; Embryo Transfer ; Embryonic Structures ; Estradiol ; Female ; Follicle Stimulating Hormone, Human* ; Gonadotropins ; Humans* ; Infertility ; Oocytes ; Ovary ; Pregnancy Rate ; Social Sciences

Antirheumatic gold compounds have been shown to inhibit NF-kB activation by blocking IkB kinase (IKK) activity. To examine the possible inhibitory mechanism of gold compounds, we expressed wild type and mutant forms of IKk alpha and beta subunits in COS-7 cells and determined the effect of gold on the activity of these enzymes both in vivo and in vitro. Substitution of Cys-179 of IKK beta with alanine (C179A) rendered the enzyme to become resistant to inhibition by a gold compound auranofin, however, similar protective effect was not observed with an equivalent level of IKK alpha (C178A) mutant expressed in the cells. Auranofin inhibited constitutively active IKK alpha and beta and variants; IKK alpha (S176E, S180E) or IKK beta (S177E, S181E), suggesting that gold directly cause inhibition of activated enzyme. The different inhibitory effect of auranofin on IKK alpha (C178A) and IKK beta (C179A) mutants indicates that gold could inhibit the two subunits of IKK in a different mode, and the inhibition of NF- kB and IKK activation induced by inflammatory signals in gold-treated cells appears through its interaction with Cys-179 of IKK beta.
Amino Acid Substitution ; Animals ; Auranofin/*pharmacology ; COS Cells ; Cysteine/genetics/*metabolism ; Enzyme Activation/drug effects ; Gold Compounds/*pharmacology ; Protein Subunits/chemistry ; Protein-Serine-Threonine Kinases/*antagonists & inhibitors/chemistry/genetics/*metabolism ; Sulfhydryl Compounds/pharmacology

I kappa B kinase beta (IKK beta) subunit of IKK complex is essential for the activation of NF-kappa B in response to various proinflammatory signals. Cys-179 in the activation loop of IKK beta is known to be the target site for IKK inhibitors such as cyclopentenone prostaglandins, arsenite, and antirheumatic gold compounds. Here we show that a mutant IKK beta in which Cys-179 is substituted with alanine had decreased activity when it was expressed in HEK-293 cells, and TNF stimulation did not restore the activity. Phosphorylation of activation loop serines (Ser-177 and Ser-181) which is required for IKK beta activation was reduced in the IKK beta (C179A) mutant. The activity of IKK beta (C179A) was partially recovered when its phosphorylation was enforced by coexpression with mitogen-activated protein kinase kinase kinases (MAPKKK) such as NF-kappa B inducing kinase (NIK) and MAPK/extracellular signal-regulated kinase kinase kinase 1(MEKK1) or when the serine residues were replaced with phospho-mimetic glutamate. The IKK beta (C179A) mutant was normal in dimer formation, while its activity abnormally responded to the change in the concentration of substrate ATP in reaction mixture. Our results suggest that Cys-179 of IKK beta plays a critical role in enzyme activation by promoting phosphorylation of activation-loop serines and interaction with ATP.
Transfection ; Serine/*metabolism ; Protein Binding ; Phosphorylation ; Mutant Proteins/chemistry ; MAP Kinase Kinase Kinases/metabolism ; I-kappa B Kinase/*chemistry ; Humans ; Hela Cells ; Enzyme Activation/*physiology ; Cysteine/*physiology ; Cells, Cultured ; Catalytic Domain ; Amino Acid Substitution/physiology ; Adenosine Triphosphate/metabolism

PURPOSE: Arachidonic acid is known to have effects on cell growth as a controller of cell proliferation and a stimulator of platelet derived growth factor. We investigated the correlations among birth weight, placental weight and free arachidonic acid level in cord blood. METHODS: Twenty full-term well babies were included in this study. Those who had sepsis, IUGR, twin that may affect the level of arachidonic acid, birth weight, and/or placental weight were excluded. Free arachidonic acid level were measured by gas chromatographic-mass spectrometry. (GC-MS) Correlation Pearson Coefficients were used for statistical analyses. RESULTS: The correlation coefficients between birth weight and placental weight, birth weight and arachidonic acid level, and placental weight and arachidonic acid level were 0.727 (P<0.001), 0.440 (P>0.05), 0.407 (P>0.05) respectively. CONCLUSION: There was statistically significant positive correlation between birth weight and placental weight. But since cord blood free arachidonic acid levels were not correlated with placental weight and birth weight, we conclude that cord blood free arachidonic acid level has no effects on fetal growth and placental growth.
Arachidonic Acid* ; Birth Weight* ; Cell Proliferation ; Fetal Blood* ; Fetal Development ; Fetal Growth Retardation ; Humans ; Parturition* ; Platelet-Derived Growth Factor ; Sepsis ; Spectrum Analysis

PURPOSE: Arachidonic acid is known to have effects on cell growth as a controller of cell proliferation and a stimulator of platelet derived growth factor. We investigated the correlations among birth weight, placental weight and free arachidonic acid level in cord blood. METHODS: Twenty full-term well babies were included in this study. Those who had sepsis, IUGR, twin that may affect the level of arachidonic acid, birth weight, and/or placental weight were excluded. Free arachidonic acid level were measured by gas chromatographic-mass spectrometry. (GC-MS) Correlation Pearson Coefficients were used for statistical analyses. RESULTS: The correlation coefficients between birth weight and placental weight, birth weight and arachidonic acid level, and placental weight and arachidonic acid level were 0.727 (P<0.001), 0.440 (P>0.05), 0.407 (P>0.05) respectively. CONCLUSION: There was statistically significant positive correlation between birth weight and placental weight. But since cord blood free arachidonic acid levels were not correlated with placental weight and birth weight, we conclude that cord blood free arachidonic acid level has no effects on fetal growth and placental growth.
Arachidonic Acid* ; Birth Weight* ; Cell Proliferation ; Fetal Blood* ; Fetal Development ; Fetal Growth Retardation ; Humans ; Parturition* ; Platelet-Derived Growth Factor ; Sepsis ; Spectrum Analysis

OBJECTIVES: Psychotic Symptom Rating Scale (PSYRATS) is an assessment tool to measure the severity of different dimensions of auditory hallucinations and delusions. The reliability and validity of the Korean version of PSYRATS (K-PSYRATS) were examined in Korean patients with major psychosis. METHODS: The inter-rater reliability of the K-PSYRATS was determined from the videotaped interviews of the five schizophrenic patients. To measure validity and internal consistency reliability, the 109 patients with auditory hallucinations or delusions were assessed using the K-PSYRATS, Positive and Negative Syndrome Scale (PANSS) and Clinical Global Impression-Severity (CGI-S) scale. RESULTS: K-PSYRATS was found to have excellent inter-rater reliability (intra-class correlation coefficient of auditory hallucination= 0.81, p<.001, intra-class correlation coefficient of delusion=0.97, p<.001) and internal consistency reliability (Cronbach's alpha of auditory hallucination=0.77, Cronbach's alpha of delusion=0.76). Significant correlation was found between K-PSYRATS and positive syndrome subscale of PANSS and CGI. CONCLUSION: K-PSYRATS is a useful assessment instrument for psychotic symptoms in Korea.
Delusions ; Hallucinations ; Humans ; Korea ; Psychotic Disorders ; Reproducibility of Results*

Differentiation of lymph node from vessels or bowel wall with similar clearity is often difficult in conventional contrast-enhanced computed tomography (CCCT). For optimal differentiation of these structures, arterial-phase dominent CT images by IV bolus injection and rapid scan were obtained. The images were compared from those by CCCT in evaluation of lymphadenopathy. Seventy patients, diagnosed as stomach cancer by pathology, were selected for this study. There were 14 cases of equivocal lymphadenopathy on CCCT while IV bolus CT revealed lymphadenopathy in 9 of there and no lymphadenopathy in the rest. Among the 38 cores without lymphadenopathy on CCCT, IV bolus CT detected 4 lymphadenopathy. In 30 node dissection cases, when we decided 1cm as CT criterion for lymphatic enlargement, the sensitivity and the specificity that there were lymphatic enlargement in gross of lymphatic infiltration above one fourth in pathology even though no lymphatic enlargement in gross were 100%. On conclusion, IV bolus CT is more helpful method to evaluate lymphadenopathy than CCCT and compatible with pathologic findings when we decide 1cm as CT criterion for lymphadenopathy. In conclusion, IV bolus CT is superior to CCCT in evaluation of metastatic Lymphadenopathy when using CT Criferion of 1cm as the diameter of enlarged lymph node.
Humans ; Lymph Nodes ; Lymphatic Diseases* ; Methods ; Pathology ; Sensitivity and Specificity ; Stomach Neoplasms* ; Stomach*

Reactive oxygen species (ROS) has been implicated as an inducer of NF-kappaB activity in numbers of cell types where exposure of cells to ROS such as H2O2 leads to NF-kappaB activation. In contrast, exposure to oxidative stress in certain cell types induced reduction of tumor necrosis factor (TNF)-induced NF-kappaB activation. And various thiol-modifying agents including gold compounds and cyclopentenone prostaglandins inhibit NF-kappaB activation by blocking IkappaB kinase (IKK). To understand such conflicting effect of oxidative stress on NF-kappaB activation, HeLa cells were incubated with H2O2 or diamide and TNF-induced expression of NF-kappaB reporter gene was measured. NF-kappaB activation was significantly blocked by these oxidizing agents, and the inhibition was accompanied with reduced nuclear NF-kappaB and inappropriate cytosolic IkappaB degradation. H2O2 and diamide also inhibited IKK activation in HeLa and RAW 264.7 cells stimulated with TNF and lipopolysaccharide, respectively, and directly blocked IKK activity in vitro. In cells treated with H2O2 alone, nuclear NF-kappaB was induced after 2 h without detectible degradation of cytosolic IkBa or activation of IKK. Our results suggest that ROS has a dual effect on NF-kappaB activation in the same HeLa cells: it inhibits acute IKK-mediated NF-kappaB activation induced by inflammatory signals, while longer-term exposure to ROS induces NF-kappaB activity through an IKK-independent pathway.
Cell Nucleus/drug effects/metabolism ; Cytosol/drug effects/metabolism ; Diamide/pharmacology ; Hela Cells/drug effects/metabolism ; Human ; Hydrogen Peroxide/pharmacology ; I-kappa B/drug effects/metabolism ; NF-kappa B/drug effects/genetics/*metabolism ; Oxidants/pharmacology ; *Oxidative Stress ; Protein-Serine-Threonine Kinases/metabolism ; Signal Transduction/drug effects ; Time Factors ; Transcription, Genetic ; Tumor Necrosis Factor/pharmacology

Direct puncture and embolization of the transverse sinus (TS) for treatment of dural arteriovenous fistula (DAVF) is typically performed with coils with or without glue. We report a case of DAVF at the left TS that was treated with Onyx embolization via direct puncture of the TS. A 75-year-old woman presented with tremor, festinating gait, and dysarthria. A left TS-DAVF with retrograde superior sagittal sinus and cortical venous reflux (Cognard type IIa+b) was identified on cerebral angiography, and both TSs were occluded with thrombi. We considered that achieving complete cure by transvenous embolization via the femoral vein or transarterial embolization via occipital feeders would be difficult. Thus, we performed a small craniotomy at the occipital bone to puncture the TS. The midportion of the TS was directly punctured with a 21-G microneedle under fluoroscopic guidance. We inserted a 5-F sheath into the TS. A microcatheter was then navigated into the affected sinus. Coils were placed through the microcatheter to support Onyx formation by reducing the pressure of shunting flow. Onyx embolization was performed with the same microcatheter. The DAVF was almost completely occluded except for the presence of minimal shunting flow to the proximal TS. After 1 week, time-of-flight magnetic resonance angiography showed complete resolution of DAVF. The patient showed resolved tremor and markedly improved mental status at 1-month follow up. Direct puncture and embolization of the TS using coils and Onyx is effective and feasible method for the treatment of DAVF when other approaches seem difficult.
Adhesives ; Aged ; Central Nervous System Vascular Malformations ; Cerebral Angiography ; Craniotomy ; Dysarthria ; Female ; Femoral Vein ; Follow-Up Studies ; Gait Disorders, Neurologic ; Humans ; Magnetic Resonance Angiography ; Methods ; Occipital Bone ; Punctures ; Superior Sagittal Sinus ; Tremor