Steric trapping reveals a cooperativity network in the intramembrane protease GlpG.

Membrane proteins are assembled through balanced interactions among proteins, lipids and water. Studying their folding while maintaining the native lipid environment is necessary but challenging. Here we present methods for analyzing key elements of membrane protein folding including thermodynamic stability, compactness of the unfolded state and folding cooperativity under native conditions. The methods are based on steric trapping, which couples the unfolding of a doubly biotinylated protein to the binding of monovalent streptavidin (mSA). We further advanced this technology for general application by developing versatile biotin probes possessing spectroscopic reporters that are sensitized by mSA binding or protein unfolding. By applying these methods to the Escherichia coli intramembrane protease GlpG, we elucidated a widely unraveled unfolded state, subglobal unfolding of the region encompassing the active site, and a network of cooperative and localized interactions to maintain stability. These findings provide crucial insights into the folding energy landscape of membrane proteins

Reduction of urate crystal-induced inflammation by root extracts from traditional oriental medicinal plants: elevation of prostaglandin D2 levels

Dried roots of the plants Acanthopanax senticosus, Angelica sinensis and Scutellaria baicalensis are used in traditional oriental medicine and reportedly possess anti-inflammatory properties. Using the murine air pouch model of inflammation, we investigated the efficacy and mode of action of an extract from these three plants in crystal-induced inflammation. Air pouches were raised on the backs of 8-week-old BALB/c mice. Mice were fed 100 mg/kg body weight of root extracts (A. senticosus:A. sinensis:S. baicalensis mixed in a ratio of 5:4:1 by weight) or vehicle only on days 3–6. Inflammation was elicited on day 6 by injecting 2 mg of monosodium urate (MSU) crystals into the pouch. Neutrophil density and IL-6 and TNF-α mRNA levels were determined in the pouch membrane, and the leukocyte count and IL-6, prostaglandin E2 (PGE2) and prostaglandin D2 (PGD2) levels were determined in the pouch exudate. Treatment with the root extracts led to a reduction in all inflammatory parameters: the leukocyte count in the pouch exudate decreased by 82%; the neutrophil density in the pouch membrane decreased by 68%; IL-6 and TNF-α mRNA levels in the pouch membrane decreased by 100%; the IL-6 concentration in the pouch fluid decreased by 50%; and the PGE2 concentration in the pouch fluid decreased by 69%. Remarkably, the concentration of the potentially anti-inflammatory PGD2 rose 5.2-fold in the pouch exudate (p < 0.005), which led to a normalization of the PGD2:PGE2 ratio. A 3.7-fold rise in hematopoietic PGD synthase (h-PGDS) mRNA paralleled this rise in PGD2 (p = 0.01)

Impact of monthly arteriovenous fistula flow surveillance on hemodialysis access thrombosis and loss

Arteriovenous fistula flow dysfunction is the leading cause of vascular access thrombosis and loss in patients undergoing hemodialysis. However, data regarding the influence of access flow rate measurements on the long-term outcomes of access are limited. This study aims to identify accesses at a high risk of thrombosis and loss among patients undergoing hemodialysis by measuring the access flow rate and exploring an optimal threshold value for predicting future access thrombosis. We enrolled 220 patients with arteriovenous fistula undergoing hemodialysis. The primary outcome was the occurrence of access thrombosis. Access flow rates were measured monthly using the ultrasound dilution method and were averaged using all measurements from patients with patent access. In patients experienced access thrombosis, those immediately before the thrombosis were selected. Using these data, we calculated the access flow rate threshold for thrombosis occurrence by analyzing the receiver operating characteristic curve, and the patients were divided into two groups according to whether access flow rates were higher or lower than 400 mL/min. During a median follow-up period of 3.1 years, 4,510 access flows were measured (median measurements per patient, 33 times; interquartile range, 11-54). A total of 65 access thromboses and 19 abandonments were observed. Access thrombosis and loss were higher in the lowflow group than in the high-flow group. This study revealed that low access flow rates are strongly associated with access thrombosis occurrence and subsequent loss of arteriovenous fistulas in patients undergoing hemodialysis

Interkulturelle Wirtschaftskommunikation zwischen Deutschland und Korea: Problematik im interkulturellen Verstehen in Bezug auf die Arbeitsweise von Deutschen und Koreanern

Economical relationships between German and South Korean companies are successfully growing. Nevertheless, difficulties in understanding each other in face-to-face-communication between managers of both countries become evident when management styles of the own culture are applied to multi-cultural business situations. Such conflicts lead to an increasing interest in theories of intercultural business communication in South Korea. The following article summarizes the outcome of a research project "A Transcultural Study for Solution of Trading Conflicts between Korea and the EU" which was conducted in 2003. The goal of the project was to make out culture-based differences in German and Korean management styles. The focus was put on three main domains which seem to be the most outstanding causes of conflicts: 1. valuation of the written and oral word in business communication; 2. concepts of time in mutual planing activities and 3. methods of operation in carrying out business plans. 30 managers of both countries were interviewed about their work experiences in the other country. Hofstede's modell of cultural dimensions and Hall's description of cultures provide the foundation for comparing and interpreting their answers. The article concludes with suggestions for improving future enterprises in the field of business between both countries

Diagnosis in a Preclinical Model of Bladder Pain Syndrome Using a Au/ZnO Nanorod-based SERS Substrate

To evaluate the feasibility of ZnO nanorod-based surface enhanced Raman scattering (SERS) diagnostics for disease models, particularly for interstitial cystitis/bladder pain syndrome (IC/BPS), ZnO-based SERS sensing chips were developed and applied to an animal disease model. ZnO nanorods were grown to form nano-sized porous structures and coated with gold to facilitate size-selective biomarker detection. Raman spectra were acquired on a surface enhanced Raman substrate from the urine in a rat model of IC/BPS and analyzed using a statistical analysis method called principal component analysis (PCA). The nanorods grown after the ZnO seed deposition were 30 to 50 nm in diameter and 500 to 600 nm in length. A volume of gold corresponding to a thin film thickness of 100 nm was deposited on the grown nanorod structure. Raman spectroscopic signals were measured in the scattered region for nanometer biomarker detection to indicate IC/BPS. The Raman peaks for the control group and IC/BPS group are observed at 641, 683, 723, 873, 1002, 1030, and 1355 cm(-1),which corresponded to various bonding types and compounds. The PCA results are plotted in 2D and 3D. The Raman signals and statistical analyses obtained from the nano-sized biomarkers of intractable inflammatory diseases demonstrate the possibility of an early diagnosis

A Case of Painful Hashimoto Thyroiditis that Mimicked Subacute Thyroiditis

Hashimoto thyroiditis (HT) is an autoimmune thyroid disorder that usually presents as a diffuse, nontender goiter, whereas subacute thyroiditis (SAT) is an uncommon disease that is characterized by tender thyroid enlargement, transient thyrotoxicosis, and an elevated erythrocyte sedimentation rate (ESR). Very rarely, patients with HT can present with painful, tender goiter or fever, a mimic of SAT. We report a case of painful HT in a 68-year-old woman who presented with pain and tenderness in a chronic goiter. Her ESR was definitely elevated and her thyroid laboratory tests suggested subclinical hypothyroidism of autoimmune origin. 99mTc pertechnetate uptake was markedly decreased. Fine needle aspiration biopsy revealed reactive and polymorphous lymphoid cells and occasional epithelial cells with Hürthle cell changes. Her clinical symptoms showed a dramatic response to glucocorticoid treatment. She became hypothyroid finally and is now on levothyroxine therapy

Development of theranostic dual-layered Au-liposome for effective tumor targeting and photothermal therapy

Abstract Background Photothermal therapy (PTT) is an emerging anti-cancer therapeutic strategy that generates hyperthermia to ablate cancer cells under laser irradiation. Gold (Au) coated liposome (AL) was reported as an effective PTT agent with good biocompatibility and excretory property. However, exposed Au components on liposomes can cause instability in vivo and difficulty in further functionalization. Results Herein, we developed a theranostic dual-layered nanomaterial by adding liposomal layer to AL (LAL), followed by attaching polyethylene glycol (PEG) and radiolabeling. Functionalization with PEG improves the in vivo stability of LAL, and radioisotope labeling enables in vivo imaging of LAL. Functionalized LAL is stable in physiological conditions, and 64Cu labeled LAL (64Cu-LAL) shows a sufficient blood circulation property and an effective tumor targeting ability of 16.4%ID g−1 from in vivo positron emission tomography (PET) imaging. Also, intravenously injected LAL shows higher tumor targeting, temperature elevation in vivo, and better PTT effect in orthotopic breast cancer mouse model compared to AL. The tumor growth inhibition rate of LAL was 3.9-fold higher than AL. Conclusion Based on these high stability, in vivo imaging ability, and tumor targeting efficiency, LAL could be a promising theranostic PTT agent. Graphic Abstrac

Connectivity between sea turtles off Jeju Island on the Korean Peninsula, and other populations in the western Pacific

The northwestern Pacific region is an important habitat for sea turtles, hosting five species out of seven. There is still limited information available about the sea turtle aggregations around the Korean Peninsula, which is the northern boundary for many sea turtle species in the western Pacific area. The present study aims to investigate the migratory route of sea turtles visiting Jeju Island. Five species of sea turtles were identified from by-catch and stranding data between 2013 and 2022 on Jeju Island in Korea: green (Chelonia mydas; 24 individuals), loggerhead (Caretta caretta; 9), hawksbill (Eretmochelys imbricata; 2), olive ridley (Lepidochelys olivacea; 2), and leatherback (Dermochelys coriacea; 1). Mixed stock analysis using mitochondrial DNA haplotypes revealed that Jeju green turtles primarily originate from the rookeries of the Japanese Archipelago. This connectivity between two regions was also supported by the similar genetic composition of loggerhead turtles. Similarly, satellite tracking data showed that several green turtles originating from Jeju Island migrated to waters near the Ryukyu Archipelago in Japan. Nevertheless, about 60% of the tracked green turtles stayed near Jeju Island, with most overwintering there, indicating the long residency in Jeju Island. This study also provides the genetic sequences of other three species including new orphan haplotypes of hawksbill and olive ridley turtles. Our findings suggest that Jeju Island serves as a stable foraging habitat and provide insight into understanding the habitat range of sea turtles in the western Pacific

Optimization of ZnO Nanorod-Based Surface Enhanced Raman Scattering Substrates for Bio-Applications

Nanorods based on ZnO for surface enhanced Raman spectroscopy are promising for the non-invasive and rapid detection of biomarkers and diagnosis of disease. However, optimization of nanorod and coating parameters is essential to their practical application. With the goal of establishing a baseline for early detection in biological applications, gold-coated ZnO nanorods were grown and coated to form porous structures. Prior to gold deposition, the grown nanorods were 30-50 nm in diameter and 500-600 nm in length. Gold coatings were grown on the nanorod structure to a series of thicknesses between 100 and 300 nm. A gold coating of 200 nm was found to optimize the Rhodamine B model analyte signal, while performance for rat urine depended on the biomarkers to be detected. These results establish design guidelines for future use of Au-ZnO nanorods in the study and early diagnosis of inflammatory diseases

Effects of stimulating interleukin -2/anti- interleukin -2 antibody complexes on renal cell carcinoma

Abstract Background Current therapies for advanced renal cell carcinoma (RCC) have low cure rates or significant side effects. It has been reported that complexes composed of interleukin (IL)-2 and stimulating anti-IL-2 antibody (IL-2C) suppress malignant melanoma growth. We investigated whether it could have similar effects on RCC. Methods A syngeneic RCC model was established by subcutaneously injecting RENCA cells into BALB/c mice, which were administered IL-2C or phosphate-buffered saline every other day for 4 weeks. RCC size was measured serially, and its weight was assessed 4 weeks after RENCA injection. Immune cell infiltration into RCC lesions and spleen was assessed by flow cytometry and immunohistochemistry. Results IL-2C treatment increased the numbers of CD8+ memory T and natural killer (NK) cells in healthy BALB/c mice (P < 0.01). In the spleen of RCC mice, IL-2C treatment also increased the number of CD8+ memory T, NK cells, and macrophages as compared to PBS-treated controls (P < 0.01). The number of interferon-γ- and IL-10-producing splenocytes increased and decreased, respectively after 4 weeks in the IL-2C-treated mice (P < 0.01). Tumor-infiltrating immune cells including CD4+ T, CD8+ T, NK cells as well as macrophages were increased in IL-2C-treated mice than controls (P < 0.05). Pulmonary edema, the most serious side effect of IL-2 therapy, was not exacerbated by IL-2C treatment. However, IL-2C had insignificant inhibitory effect on RCC growth (P = 0.1756). Conclusions IL-2C enhanced immune response without significant side effects; however, this activity was not sufficient to inhibit RCC growth in a syngeneic, murine model