RHS6 coordinately regulates the Th2 cytokine genes by recruiting GATA3, SATB1, and IRF4.

BACKGROUND: Asthma is a Th2 cell-driven inflammatory disease and a major public health concern. The cis-acting element Rad50 hypersensitive site 6 (RHS6) in the Th2 locus control region is essential for regulation of the Th2 cytokine genes; however, its role in allergic airway inflammation and underlying molecular mechanisms of the regulation by RHS6 are poorly understood. OBJECTIVE: We sought to understand the role of RHS6 in the development of allergic airway inflammation and its molecular mechanism for Th2 cytokine expression. METHODS: We used an ovalbumin-induced allergic inflammation model with RHS6-deficient mice to examine the role of RHS6 in this process. To examine molecular mechanism of RHS6 for Th2 cytokine expression, we used DNA affinity chromatography and mass spectrometry, quantitative RT-PCR, ELISA, intracellular cytokine staining, chromatin immunoprecipitation, and co-immunoprecipitation. RESULTS: Deletion of RHS6 caused a dramatic resistance to allergic airway inflammation. RHS6 recruited transcription factors GATA3, SATB1, and IRF4, which play important roles in expression of all three Th2 cytokine genes. RHS6 deficiency caused inhibition of transcription factor-induced Th2 cytokine gene expression. CONCLUSION: RHS6 is a critical regulatory element for allergic airway inflammation and for coordinate regulation of Th2 cytokine genes by recruiting GATA3, SATB1, and IRF4.

Indicative factors for surgical or angiographic intervention in hemodynamically stable patients with blunt abdominal trauma: A retrospective cohort study.

INTRODUCTION: The standard of care for intraperitoneal injury in hemodynamically stable patients after blunt abdominal trauma has been replaced by non-operative management (NOM). However, selective NOM, depending on the situation, seems necessary in determining the treatment plan. In this study, we attempted to identify risk factors for surgical or angiographic intervention (SAI) in hemodynamically stable blunt abdominal trauma patients. METHODS: This retrospective study which included adult patients who were brought to a regional trauma center was conducted from March 2015 to October 2019. We evaluated the characteristics of blunt abdominal trauma patients and analyzed factors that were related to the requirement of SAI in these patients. Patients were divided into SAI and conservative management (CM) groups. RESULTS: We reviewed 1,176 patients, and after exclusions, of whom 248 blunt abdominal trauma and free fluid observed on CT were identified. The mean pulse rate was higher in the SAI than in the CM (P=0.025). Laboratory findings showed that lactate and delta neutrophil index (DNI) levels were higher in the SAI than in the CM (P=0.002 and 0.026 respectively). Additionally, the mean free fluid size in the SAI (85.69mm) was significantly larger than that in the CM (68.12mm; P=0.001), and blush was more frequently observed in the SAI (P<0.001). In multivariate analysis, only blush was an independent prognostic factor for SAI (OR 11.7, 95% CI, 5.1-30.8, P<0.001). CONCLUSION: In hemodynamically stable patients with blunt abdominal trauma, blush but also high lactate and DNI are associated with the requirement of interventional radiology and/or surgery.

Are the Oxford(®) medial unicompartmental knee arthroplasty new instruments reducing the bearing dislocation risk while improving components relationships? A case control study.

INTRODUCTION: The recently introduced Microplasty(®) system is an upgrade of conventional phase III instrumentation. However, little is known of its impact on the position of the implant following the Oxford(®) mobile-bearing unicompartmental knee arthroplasty (UKA). This study investigated whether the Microplasty(®) instrumentation system can improve the positioning of the implant and reduce the rate of early failure. HYPOTHESIS: Microplasty(®) provides a better positioning and decreases the rate of dislocation. MATERIALS AND METHODS: The medical records and radiographs of 82 consecutive Oxford(®) UKAs were reviewed retrospectively. The radiographic parameters and prevalence of early failure of 41 UKAs performed with the Microplasty(®) system and 41 UKAs using the conventional instrumentation system were compared. Both groups were comparable in terms of demographics and preoperative status. RESULTS: The femoral components in the Microplasty(®) group were more contiguously placed and more convergent in relation to the tibial components compared to the conventional instrumentation system (P<0.01). The frequency of bearing dislocation was lower in the Microplasty(®) group (P=0.04). A wide gap and the angle between components were associated with an increased risk of bearing dislocation. CONCLUSION: The Microplasty(®) instrumentation system consistently placed the femoral and tibial components in more contiguous and convergent positions. Such changes in position decreased the risk of bearing dislocations by reducing the space available for bearing rotation. LEVEL OF EVIDENCE: Level III, case control study.

The fouling characterization and control in the high concentration PAC membrane bioreactor HCPAC-MBR.

This study focuses on the experimental investigation to identify the effect of PAC at high concentrations on the fouling of membranes. A pilot-scale experimental apparatus was installed at a water treatment plant located downstream of Nakdong river basin, Korea. Effluent of rapid sand filter was used as influent of the system, which consists of PAC bio-reactor, submerged membrane module (hollow fiber with pore size 0.1 m) and air supply facility. PAC was dosed at 40 g/L initially and it was not replaced during the operation period. Suction type filtration was carried out at intervals of 12 min. suction and 3 min. idling. At the initial flux 0.36 m/d, the system could be operated stably for around 90 days at target trans-membrane pressure (TMP) of 40 kPa. Among total resistance of membrane filtration, cake and gel layer resistance, Rc+Rg, was the dominant fraction (more than 90% of the total) to increase the filtration pressure, which means that the filtration resistance could be controlled by the PAC cake layer and then irreversible membrane fouling could be prevented. Three minutes air backwashing every 3 days could extend the operation period to 127 days. Organics were analyzed in terms of molecular weight structure. The influent of the system consists of 15.0% and 74.4% of hydrophobic and hydrophilic natural organic matter (NOM), respectively. Hydrophobic and hydrophilic (electrostatic) interaction was the main factor on fouling of the membrane in the reactor. Hydrophobic fraction decreased slightly in the effluent, which means hydrophobic NOM removal in the reactor by adsorption. Organics accumulated in the membrane were extracted for analysis after a certain period of operation. The fraction of hydrophobic and hydrophilic organics was 41.4% and 38.9%, respectively. On the basis of the experimental results, the hydrophobic organics were the major materials causing the fouling of the membrane, which should be changed to other types of material.

The role of sLZIP in cyclin D3-mediated negative regulation of androgen receptor transactivation and its involvement in prostate cancer.

Androgen and the androgen receptor (AR) have important roles in prostate cancer (PCa) development, and androgen ablation has been the main therapeutic option for the treatment of PCa. However, the transition mechanism from androgen-dependent to -independent PCa after androgen depletion remains unclear. We investigated the distinct roles of small leucine zipper protein (sLZIP) in proliferation of androgen-dependent and -independent PCa cells. Cyclin D3 is known to interact with AR and attenuates the ligand-dependent function of AR in PCa cells. sLZIP regulates the transcription of cyclin D3 by binding directly to the AP-1 region in the cyclin D3 promoter. sLZIP represses AR transcriptional activity by interaction with AR that is phosphorylated by cyclin D3/cyclin-dependent kinase11(p58), leading to the suppression of androgen-dependent proliferation of PCa cells. The expression level of sLZIP is elevated in androgen-independent PCa cells and advanced human prostate tumors. Knockdown of endogenous sLZIP suppresses proliferation of androgen-independent PCa cells. LNCaP cells transformed to androgen-independent PCa cells exhibit increased expressions of sLZIP and cyclin D3. Tumor formation is inhibited in nude mouse xenografts from two androgen-independent PCa cells that are stably transfected with sh-sLZIP. Our findings indicate that sLZIP negatively regulates AR transactivation in androgen-dependent PCa cells and functions as a positive regulator in tumor progression of androgen-independent PCa. sLZIP contributes to the malignant phenotype of PCa and constitutes a novel therapeutic target for human PCa.

Comparative bioavailability of silibinin in healthy male volunteers.

AIM: To study a comparative bioavailability of Liverman capsule to Legaion capsule and Silymarin tablet (which contain silibinin) in 24 healthy volunteers. VOLUNTEERS AND METHODS: Twenty-four healthy male Korean volunteers received each medicine at the silibinin dose of 120 mg in a 3 x 3 crossover study. There was a 1-week washout period among the doses. Plasma concentrations of silibinin were monitored by a high-performance liquid chromatography for over a period of 12 hours after the administration. AUCinf (the area under the plasma concentration-time curve from time zero to time infinity) was calculated by the trapezoidal rule extrapolation method. Cmax (maximum plasma drug concentration) and tmax (time to reach a Cmax) were compiled from the plasma concentration-time data. Analysis of variance was carried out using logarithmically transformed AUCinf, AUC(0-12h), and Cmax and untransformed tmax. RESULTS: After an oral administration of Liverman capsule, the pharmacokinetic parameters of silibinin, such as AUC(0-12h) (5.59, 4.24 and 13.9 microg/ml x h for Legalon capsule, Silymarin tablet and Liverman capsule, respectively) and AUCinf (6.00, 4.63 and 15.1 microg/ml x h) were significantly greater, Cmax (1.33, 1.13 and 6.04 microg/ml) was significantly higher and tmax (1.83, 2.10 and 0.875 h) was significantly faster than those after Legalon capsule and Silymarin tablet. CONCLUSION: These results indicate that the absorption and the extent of relative oral bioavailability of silibinin after Liverman capsule were significantly faster and greater, respectively, than those after Legalon capsule and Silymarin tablet.

Design and fabrication of a superconducting magnet for an 18 GHz electron cyclotron resonance ion∕photon source NFRI-ECRIPS.

A superconducting magnet was designed and fabricated for an 18 GHz ECR ion∕photon source, which will be installed at National Fusion Research Institute (NFRI) in South Korea. The magnetic system consists of a set of four superconducting coils for axial mirror field and 36 pieces of permanent magnets for hexapolar field. The superconducting coils with a cryocooler (1.5 W @ 4.2 K) allow one to reach peak mirror fields of 2.2 T in the injection and those of 1.5 T in the extraction regions on the source axis, and the resultant hexapolar field gives 1.35 T on the plasma chamber wall. The unbalanced magnetic force between the coils and surrounding yoke has been minimized to 16 ton by a coil arrangement and their electrical connection, and then was successfully suspended by 12 strong thermal insulating supports made of large numbers of carbon fibers. In order to block radiative thermal losses, multilayer thermal insulations are covered on the coil windings as well as 40-K aluminum thermal shield. Also new schemes of quench detection and safety system (coil divisions, quench detection coils, and heaters) were employed. For impregnation of the windings a special epoxy has been selected and treated to have a higher breaking strength and a higher thermal conductivity, which enables the superconductors to be uniformly and rapidly cooled down or heated during a quench.

Ebp1 sumoylation, regulated by TLS/FUS E3 ligase, is required for its anti-proliferative activity.

Ebp1, an ErbB3 receptor-binding protein, inhibits cell proliferation and acts as a putative tumor suppressor. Ebp1 translocates into the nucleus and functions as a transcription co-repressor for E2F-1. Here, we show that Ebp1 p42 isoform can be sumoylated on both K93 and K298 residues, which mediate its nuclear translocation and are required for its anti-proliferative activity. We find that translocation in liposarcoma (TLS)/FUS, an RNA-binding nuclear protein that is involved in pre-mRNA processing and nucleocytoplasmic shuttling, has Sumo1 E3 ligase activity for Ebp1 p42. Ebp1 directly binds TLS/FUS, which is regulated by genotoxic stress-triggered phosphorylation on Ebp1. Ebp1 sumoylation facilitates its nucleolar distribution and protein stability. Overexpression of TLS enhances Ebp1 sumoylation, whereas depletion of TLS abolishes Ebp1 sumoylation. Moreover, unsumoylated Ebp1 mutants fail to suppress E2F-1-regulated transcription, resulting in loss of its anti-proliferation activity. Hence, TLS-mediated sumoylation is required for Ebp1 transcriptional repressive activity.

NGF inhibits human leukemia proliferation by downregulating cyclin A1 expression through promoting acinus/CtBP2 association.

Cyclin A1 is essential for leukemia progression, and its expression is tightly regulated by acinus, a nuclear speckle protein. However, the molecular mechanism of how acinus mediates cyclin A1 expression remains elusive. Here we show that transcription corepressor CtBP2 directly binds acinus, which is regulated by nerve growth factor (NGF), inhibiting its stimulatory effect on cyclin A1, but not cyclin A2, expression in leukemia. NGF, a cognate ligand for the neurotrophic receptor TrkA, promotes the interaction between CtBP2 and acinus through triggering acinus phosphorylation by Akt. Overexpression of CtBP2 diminishes cyclin A1 transcription, whereas depletion of CtBP2 abolishes NGF's suppressive effect on cyclin A1 expression. Strikingly, gambogic amide, a newly identified TrkA agonist, potently represses cyclin A1 expression, thus blocking K562 cell proliferation. Moreover, gambogic amide ameliorates the leukemia progression in K562 cells inoculated nude mice. Hence, NGF downregulates cyclin A1 expression through escalating CtBP2/acinus complex formation, and gambogic amide might be useful for human leukemia treatment.

Proteomic analysis of differential protein expression in atherosclerosis.

Although recent studies have shown that several pro-inflammatory proteins can be used as biomarkers for atherosclerosis, the mechanism of atherogenesis is unclear and little information is available regarding proteins involved in development of the disease. Atherosclerotic tissue samples were collected from patients in order to identify the proteins involved in atherogenesis. The protein expression profile of atherosclerosis patients was analysed using two-dimensional electrophoresis-based proteomics. Thirty-nine proteins were detected that were differentially expressed in the atherosclerotic aorta compared with the normal aorta. Twenty-seven of these proteins were identified in the MS-FIT database. They are involved in a number of biological processes, including calcium-mediated processes, migration of vascular smooth muscle cells, matrix metalloproteinase activation and regulation of pro-inflammatory cytokines. Confirmation of differential protein expression was performed by Western blot analysis. Potential applications of the results include the identification and characterization of signalling pathways involved in atherogenesis, and further exploration of the role of selected identified proteins in atherosclerosis.

Electrocardiographic findings of organophosphate intoxication in emergency department as predictors of prognosis: a retrospective analysis.

To investigate the clinical significance of electrocardiographic (ECG) findings in patients with organophosphate intoxication and to find predictors for prognosis, we reviewed 170 patients whose ECG was taken in the emergency department (ED) from 1981 to 1989. There were 67 cases whose ECG showed prolongation of corrected Q-T interval (QTc). In this group, the mortality rate, respiratory failure rate and frequency of ventricular premature contraction (VPC) were significantly higher than those of patients without QTc prolongation. In the group with presence of VPC, the overall mortality rate and respiratory failure rate were also significantly higher when compared with those without VPC. We concluded that QTc prolongation and the presence of VPCs in patients with organophosphate intoxication may predict their clinical respiratory failure and mortality.

Identification of Cdc6 protein domains involved in interaction with Mcm2 protein and Cdc4 protein in budding yeast cells.

The Cdc6 protein (Cdc6p) has essential roles in regulating initiation of DNA replication. Cdc6p is recruited to origins of replication by the origin recognition complex (ORC) late in mitosis; Cdc6p in turn recruits minichromosome maintenance (Mcm) proteins to form the pre-replicative complex. Cdc6p is thought to interact with one or more Mcm proteins but this point has not yet been demonstrated. In the present study we observed that Cdc6p interacted significantly only with Mcm2p out of six Mcm proteins in yeast two-hybrid cells. Our results indicate that the interaction of Cdc6p with Mcm2p is specific, although we cannot exclude the possibility that the interaction might not be direct. In attempts to identify domains of Cdc6p important for interaction with Mcm2p, we tested interactions of various deleted versions of Cdc6p with Mcm2p and also with Cdc4p, which was previously known to interact with Cdc6p. The portion of Cdc6p from amino acid residues 51 to 394 was able to interact with Mcm2p. During the course of the studies we also discovered a previously undetected Cdc4p interaction domain between residues 51 and 394. Interestingly, when all six putative Cdc28 phosphorylation sites in Cdc6p were changed to alanine, a 6-7-fold increase in binding to Mcm2p was observed. This result suggests that unphosphorylated Cdc6p has higher affinity than phosphorylated Cdc6p for Mcm2p; this might partly explain the previous observation that Cdc6p failed to load Mcm proteins on replication origins during S phase when the cyclin-dependent protein kinase was active, thus helping to prevent the reinitiation of activated replicons.

Stability of Octastatin, a somatostatin analog cyclic octapeptide, in aqueous solution.

In this study, the degradation of Octastatin, a cyclic octapeptide analog of somatostatin, was examined as a function of pH, temperature, buffer, and ionic strength by reversed-phase gradient high-performance liquid chromatography. Degradation of Octastatin followed a first-order kinetics. Various buffer species such as acetate, ammonium acetate, citrate, glutamate, phosphate, and borate showed differing effects on the degradation of the octapeptide. Good stability was found in glutamate and acetate buffer of pH 4.0. Degradation of Octastatin was greater in citrate- or phosphate-containing buffers than in glutamate or acetate buffers. With phosphate buffer, higher buffer concentration caused greater degradation, while in acetate buffer, the effect of buffer concentration and ionic strength was negligible. In addition, the degradation of Octastatin was markedly inhibited by increasing the concentration of glutamate buffer. This study allows the prediction of good stability in acetate buffer (0.01 M, pH 4.0) with a t90% of 84.1 days at 20 degrees C.

QTc prolongation indicates a poor prognosis in patients with organophosphate poisoning.

This study reviewed emergency department electrocardiograms of 223 patients with organophosphate poisoning from January 1982 to June 1994: 97 (43.5%) had QTc prolongation and were placed in group A; 126 patients without QTc prolongation were designated as group B. Compared with group B, group A patients had a higher mortality (19.6% v 4.8%, P < .001) and a higher incidence of respiratory failure (56.7% v 20.6%, P < .001). Serum cholinesterase levels were determined in the 223 patients: 92 (41.3%) were classified as severe poisoning. 32 (14.3%) as moderate, 41 (18.1%) as mild, and 58 (25.7%) as very mild. The severe group had a high incidence of QTc prolongation (P < .001), a high incidence of respiratory failure (P < 0.001), and a higher mortality rate (P < 0.001) than the other groups. Of the QTc prolongation patients, 59.8% (55/92) had a high incidence of respiratory failure (78.2% v 35.1%, P < .0001) and a higher mortality rate (29.1% v 8.1%, P < 0.05) compared with 40.2% (37/92) of the patients without QTc prolongation in the severe group. In conclusion, a complete electrocardiogram at the emergency department is important and of prognostic value.

[Clinical experience of automated double filtration plasmapheresis].

Double filtration plasmapheresis, one kind of fractionation plasmapheresis, was developed from membrane type plasmapheresis to remove only the pathogen and return the normal protein back to the patient. We started our automated double filtration plasmapheresis since December 1993. There were 13 patients who received one hundred treatments totally during one year period. And they are myasthenia gravis (8 patients); acute inflammatory demyelinating polyneuropathy (1 patient), multiple myeloma (1 patient); acquired factor VIII inhibitor (1 patient); autoimmune hemolytic anemia (1 patient); systemic lupus erythematous (1 patient). Technically double filtration plasmapheresis is easy to perform and time-saving. It also makes necessity of replacement fluid less frequent. Incidence of complication is rare, and this includes hypotension 2%, palpitation 1%, headache 1%, hemolysis 4%, air emboli 1%, high secondary pressure 2%, and no motality during our treatment. Clinical response is documented in cases of myasthenia gravis; acute inflammatory demyelinating polyneuropathy and acquired factor VIII inhibitor in our study. In conclusion, double filtration plasmapheresis is a time-saving, convenient, and safe therapeutic modality with rare complication. Because its effectiveness on limited kinds of diseases and costs relatively high price, thus plasmapheresis should be used in selected cases and treat aggressively if indicated.