Oral administration of 1,4-aryl-2-mercaptoimidazole inhibits T-cell proliferation and reduces clinical severity in the murine experimental autoimmune encephalomyelitis model.

T cells play a pivotal role in the initiation and progression of multiple sclerosis. We have found that 1,4-aryl-2-mercaptoimidazole (KRM-III) inhibited T-cell antigen receptor- and phorbol myristate acetate/ionomycin-induced activation of nuclear factor of activated T cells (NFAT) and T-cell proliferation with an IC(50) of 5 microM. The KRM-III-mediated inhibitory effect was specific for NFAT activation but not for nuclear factor kappaB. Oral administration of 90 mg/kg KRM-III resulted in complete abrogation of anti-CD3 antibody-induced T-cell activation and a 45.8% reduction in footpad swelling in bovine serum albumin-induced delayed-type hypersensitivity. In the murine experimental autoimmune encephalomyelitis (EAE) model, oral administration of KRM-III significantly attenuated the severity of disease when given before or after disease onset. Draining lymph node cells from KRM-III-treated mice showed markedly reduced proliferation in response to myelin oligodendrocyte glycoprotein peptide. Histological analysis indicated that KRM-III reduced the infiltration of inflammatory cells to the white matter of spinal lumbar cords. These results demonstrate that KRM-III efficiently inhibits T-cell activation and inflammatory responses and lessens EAE clinical signs, which suggest KRM-III as a potential lead compound for the treatment of T-cell-driven autoimmune diseases.

Reduced Tacrolimus Trough Level Is Reflected by Estimated Glomerular Filtration Rate (eGFR) Changes in Stable Renal Transplantation Recipients: Results of the OPTIMUM Phase 3 Randomized Controlled Study.

BACKGROUND Minimizing the tacrolimus dosage in patients with stable allograft function needs further investigation. MATERIAL AND METHODS We performed an open-label, randomized, controlled study from 2010 to 2016 in 7 tertiary teaching hospitals in Korea and enrolled 345 kidney transplant recipients with a stable graft status. The study group received reduced-dose tacrolimus, 1080-1440 mg/day of enteric-coated mycophenolate sodium (EC-MPS), and corticosteroids. The control group received the standard tacrolimus dosage and 540-720 mg/day of EC-MPS with steroids. The primary endpoint was the mean estimated glomerular filtration rate (eGFR) and change in the eGFR at 12 months after randomization. RESULTS The mean tacrolimus trough level of the study group was 4.51±1.62 ng/mL, which was lower than that of the control group, at 6.75±2.82 ng/mL (P<0.001). The primary endpoint was better in the study group in terms of change in eGFR (P<0.001). The month 12 eGFRs were 73.6±28.4 and 68.3±18.1 mL/min/1.73 m² in the study and the control groups, respectively, but the difference did not reach statistical significance (P=0.07). The incidence of adverse events was similar between the study and the control groups. CONCLUSIONS Minimizing tacrolimus to a trough level below 5 ng/mL combined with conventional EC-MPS can be considered in patients with a steady follow-up, as it was associated with small benefits in the changes of the eGFR (Clinicaltrials.gov number: NCT01159080).

Molecular epidemiology of community-associated antimicrobial-resistant Staphylococcus aureus in Seoul, Korea (2003): pervasiveness of multidrug-resistant SCCmec type II methicillin-resistant S. aureus.

There is an extremely high incidence of antimicrobial resistance of the clinical isolates of Staphylococcus aureus in Korea. This study carried out a molecular investigation to determine the prevalence of the community-associated antimicrobial-resistant S. aureus and methicillin-resistant S. aureus (MRSA). The percentage resistance from the nasal swabs of healthy volunteers in 2003 in Seoul is as follows: penicillin (91%), erythromycin (EM, 14%), gentamicin (GM, 9.3%), tetracycline (TE, 8.2%), cephalothin (4%), oxacillin (OX, MRSA; 3.8%), clindamycin (CC, 2.6%), ciprofloxacin (CIP, 0.8%), and sulfamethoxazole/trimethoprim (0.6%). The community-associated MRSA (C-MRSA) strains were examined by pulsed-field gel electrophoresis (PFGE) analysis of the SmaI macro-fragments, multilocus sequence typing (MLST), and staphylococcal cassette chromosome mec (SCCmec) typing using the PCR analysis. The Korean C-MRSA isolates were clustered into three distinct groups. One PFGE group containing the C-MRSA strains showed resistance to CC, EM, and GM, a high level (32-96 microg/ml) of resistance to methicillin, sequence type 5 (ST5), and SCCmec type II, which is the most common hospital associated-MRSA (H-MRSA) isolated in Korea. These results highlight the heterogeneous genetic background of the C-MRSA as well as the pervasiveness of the H-MRSA isolates in this community.

Physicochemical interactions of cycloamylose with phenolic compounds.

The complex formation capability of cycloamylose (CA), having a degree of polymerization of 23-45, with phenolic compounds (PCs) was investigated using various physicochemical techniques. The fluorescence intensity of PCs increased and then reached a plateau at 10-20mM cyclodextrin, while it continued to increase at up to 60mM CA. Thermodynamic data of CA complexes with PCs revealed that the binding process was primarily enthalpy-driven and spontaneous. CA favored to form the most stable complex with chlorogenic acid (CHA) among all PCs. Chemical shift changes for the protons in interior and exterior of CA, as well as in PCs suggested a possible formation of both inclusion and extramolecular interactions between CA and PCs. The ROESY spectrum confirmed that the aromatic moieties of CHA were partially interacted with CA molecules through relatively weak binding. XRD, DSC, and SEM results also supported the complex formation by intermolecular interaction between CA and CHA.

Novel formulation of low-fat spread using rice starch modified by 4-α-glucanotransferase.

Low-fat spreads were developed using a thermoreversible gelling agent, the 4-α-glucanotransferase (4αGT)-modified rice starch. The low-fat spreads consisted of the modified starch paste (or rice starch or maltodextrin), olive oil (0-30% w/w), egg yolk, salt, xanthan gum, and butter flavor, and were produced by homogenization, ultrasonic processing at 50% amplitude for 2min, and cold-gel setting at 4°C for 24h. Formulations with 15% and 20% of the modified starch paste resulted in highly stable oil-in-water low-fat spreads having varied textural properties and acceptable spreadability, whereas formulations with rice starch and maltodextrin did not yield enough stability and consistency. Moreover, the modified starch-based low-fat spreads showed high thermoreversibility. These results indicated that 4αGT-modified starch could be used in the preparation of low-fat spreads, allowing the formulation of functional products for healthy diets.

Physicochemical functionality of 4-α-glucanotransferase-treated rice flour in food application.

The physicochemical properties of 4-α-glucanotransferase (4αGTase)-modified rice flours were examined by measuring the molecular weight distribution, moisture sorption isotherm, and melting enthalpy of ice crystals. The results obtained by measuring the moisture sorption isotherm and melting enthalpy of ice crystals revealed that 4αGTase-modified rice flours had high water binding capacity than that of control rice flour. When the textural properties of noodles containing 4αGTase-treated rice flours after freeze-thaw cycling were measured by texture profile analysis, the textural properties of control noodle deteriorated. However, those of noodle with 4αGTase-modified rice flours were retained. For the melting enthalpy of ice crystals formed within cooked noodles, 4αGTase-treated rice flour showed similar effect to sucrose for reducing the melting enthalpy of ice crystals, however, the texture and taste of noodle with sucrose was undesirable for consuming. 4αGTase-treated rice flour appeared to have good potential as a non-sweet cryoprotectant of frozen product.

Development of reduced-fat mayonnaise using 4alphaGTase-modified rice starch and xanthan gum.

In this study a disproportionating enzyme, 4-alpha-glucanotransferase (4alphaGTase), was used to modify the structural properties of rice starch to produce a suitable fat substitute in reduced-fat (RF) mayonnaise. The mayonnaise fat was partially substituted with the 4alphaGTase-treated starch paste at levels up to 50% in combination with xanthan gum and the physical and rheological properties of the modified RF mayonnaise samples were investigated. All mayonnaises prepared in this study exhibited shear thinning behavior and yield stress. Viscoelastic properties of mayonnaise were characterized using dynamic oscillatory shear test and it was observed that mayonnaises exhibited weak gel-like properties. The magnitude of elastic and loss moduli was also affected by 4alphaGTase-treated starch concentration and presence of xanthan gum. In relation to microstructure, RF mayonnaise prepared with 3.8 or 5.6 wt% of 4alphaGTase-treated starch and xanthan gum showed smaller droplets. The use of 5.6 wt% of 4alphaGTase-treated starch and 0.1 wt% of xanthan gum produced a RF mayonnaise with similar rheological properties and appearances as FF mayonnaise with gum. This study demonstrated a high feasibility for using 4alphaGTase-treated rice starch as a viable fat replacer in mayonnaise.

Evaluation of viral clearance in the production of HPV-16 L1 virus-like particles purified from insect cell cultures.

Biopharmaceutical products produced from cell cultures have a potential for viral contamination from cell sources or from adventitious introduction during production. The objective of this study was to assess viral clearance in the production of insect cell-derived recombinant human papillomavirus (HPV)-16 type L1 virus-like particles (VLPs). We selected Japanese encephalitis virus (JEV), bovine viral diarrhea virus (BVDV), and minute virus of mice (MVM) as relevant viruses to achieve the aim of this study. A downstream process for the production of purified HPV-16 L1 VLPs consisted of detergent lysis of harvested cells, sonication, sucrose cushion centrifugation, and cesium chloride (CsCl) equilibrium density centrifugation. The capacity of each purification/treatment step to clear viruses was expressed as reduction factor by measuring the difference in log virus infectivity of sample pools before and after each process. As a result, detergent treatment (0.5% v/v, Nonidet P-40/phosphate-buffered saline) was effective for inactivating enveloped viruses such as JEV and BVDV, but no significant reduction (< 1.0 log(10)) was observed in the non-enveloped MVM. The CsCl equilibrium density centrifugation was fairly effective for separating all three relevant adventitious viruses with different CsCl buoyant density from that of HPV-16 L1 VLPs (JEV, BVDV, and MVM = 4.30, 3.10, > or = 4.40 log(10) reductions). Given the study conditions we used, overall cumulative reduction factors for clearance of JEV, BVDV, and MVM were > or = 10.50, > or = 9.20, and > or = 6.40 log(10) in 150 ml of starting cell cultures, respectively.

Two new neolignans from the aerial parts of Rodgersia podophylla.

Two new neolignans (1, 2) were isolated from the aerial parts of Rodgersia podophylla, along with four known neolignans, and their structures were elucidated using spectral experiments and comparison with literature data.

Two new acylated iridoid glucosides from the aerial parts of Paederia scandens.

Two new acylated iridoid glucosides were isolated from the aerial parts of Paederia scandens along with six known iridoid glucosides. The structures of two new compounds were elucidated as 6'-O-E-feruloylmonotropein (1) and 10-O-E-feruloylmonotropein (2) by spectroscopic methods.

Development of real-time RT-PCR for evaluation of JEV clearance during purification of HPV type 16 L1 virus-like particles.

Insect cell culture has greatly increased in part due to the widespread use of insect virus-based vectors for efficient expression of foreign proteins. Insect cells such as Sf9 cells are susceptible to arboviruses which may pose a safety concern by adventitious introduction during the production process. The objective of this study was to establish techniques for viral clearance validation of insect cell-derived biotechnological products using Japanese encephalitis virus (JEV) as a model, since JEV is a member of arthropod-borne flaviviruses that are known to be infectious in insect cells. Here we report the development of a quantitative assay for JEV RNA using real-time reverse transcription-polymerase chain reaction (RT-PCR). The assay was performed using LightCycler and RNA amplification kit SYBR Green I. The JEV specific primer was selected from the 3' untranslated region, and the expected band size was 323 base pairs (bp). The sensitivity of the assay was calculated to be approximately 15 TCID(50)per reaction. Highly reproducible standard curves were obtained from experiments performed on three different days. JEV clearance was determined during the purification process of rHPV-16 L1 VLPs by CsCl equilibrium density centrifugation. The comparative results obtained by real-time RT-PCR assay for JEV and infectivity titrations suggested that the real-time RT-PCR assay could have an additive effect on the interpretation and evaluation of virus clearance, especially during the virus removal process.