HMGB1, a potential regulator of tumor microenvironment in KSHV-infected endothelial cells

High-mobility group box 1 (HMGB1) is a protein that binds to DNA and participates in various cellular processes, including DNA repair, transcription, and inflammation. It is also associated with cancer progression and therapeutic resistance. Despite its known role in promoting tumor growth and immune evasion in the tumor microenvironment, the contribution of HMGB1 to the development of Kaposi’s sarcoma (KS) is not well understood. We investigated the effect of HMGB1 on KS pathogenesis using immortalized human endothelial cells infected with Kaposi’s sarcoma-associated human herpes virus (KSHV). Our results showed that a higher amount of HMGB1 was detected in the supernatant of KSHV-infected cells compared to that of mock-infected cells, indicating that KSHV infection induced the secretion of HMGB1 in human endothelial cells. By generating HMGB1 knockout clones from immortalized human endothelial cells using CRISPR/Cas9, we elucidated the role of HMGB1 in KSHV-infected endothelial cells. Our findings indicate that the absence of HMGB1 did not induce lytic replication in KSHV-infected cells, but the cell viability of KSHV-infected cells was decreased in both 2D and 3D cultures. Through the antibody array for cytokines and growth factors, CXCL5, PDGF-AA, G-CSF, Emmprin, IL-17A, and VEGF were found to be suppressed in HMGB1 KO KSHV-infected cells compared to the KSHV-infected wild-type control. Mechanistically, phosphorylation of p38 would be associated with transcriptional regulation of CXCL5, PDGF-A and VEGF. These observations suggest that HMGB1 may play a critical role in KS pathogenesis by regulating cytokine and growth factor secretion and emphasize its potential as a therapeutic target for KS by modulating the tumor microenvironment

Smart contact lens and transparent heat patch for remote monitoring and therapy of chronic ocular surface inflammation using mobiles

Wearable electronic devices that can monitor physiological signals of the human body to provide biomedical information have been drawing extensive interests for sustainable personal health management. Here, we report a human pilot trial of a soft, smart contact lens and a skin-attachable therapeutic device for wireless monitoring and therapy of chronic ocular surface inflammation (OSI). As a diagnostic device, this smart contact lens enables real-time measurement of the concentration of matrix metalloproteinase-9, a biomarker for OSI, in tears using a graphene field-effect transistor. As a therapeutic device, we also fabricated a stretchable and transparent heat patch attachable on the human eyelid conformably. Both diagnostic and therapeutic devices can be incorporated using a smartphone for their wireless communications, thereby achieving instantaneous diagnosis of OSI and automated hyperthermia treatments. Furthermore, in vivo tests using live animals and human subjects confirm their good biocompatibility and reliability as a noninvasive, mobile health care solution

Chemical crosslinking and mass spectrometry studies of the structure and dynamics of membrane proteins and receptors.

Membrane proteins make up a diverse and important subset of proteins for which structural information is limited. In this study, chemical cross-linking and mass spectrometry were used to explore the structure of the G-protein-coupled photoreceptor bovine rhodopsin in the dark-state conformation. All experiments were performed in rod outer segment membranes using amino acid 'handles' in the native protein sequence and thus minimizing perturbations to the native protein structure. Cysteine and lysine residues were covalently cross-linked using commercially available reagents with a range of linker arm lengths. Following chemical digestion of cross-linked protein, cross-linked peptides were identified by accurate mass measurement using liquid chromatography-fourier transform mass spectrometry and an automated data analysis pipeline. Assignments were confirmed and, if necessary, resolved, by tandem MS. The relative reactivity of lysine residues participating in cross-links was evaluated by labeling with NHS-esters. A distinct pattern of cross-link formation within the C-terminal domain, and between loop I and the C-terminal domain, emerged. Theoretical distances based on cross-linking were compared to inter-atomic distances determined from the energy-minimized X-ray crystal structure and Monte Carlo conformational search procedures. In general, the observed cross-links can be explained by re-positioning participating side-chains without significantly altering backbone structure. One exception, between C3 16 and K325, requires backbone motion to bring the reactive atoms into sufficient proximity for cross-linking. Evidence from other studies suggests that residues around K325 for a region of high backbone mobility. These findings show that cross-linking studies can provide insight into the structural dynamics of membrane proteins in their native environment

Effect of corn gluten and its hydrolysate consumptions on weight reduction in rats fed a high-fat diet

This study examined the effects of corn gluten (CG) and its hydrolysate consumptions on weight reduction in rats fed a high-fat diet. Eight-month-old male Sprague-Dawley rats (n=40) were fed a high-fat diet (40% calorie as fat) for 4 weeks. They were then randomly divided into four groups and fed the isocaloric diets with different protein sources for 8 weeks. The protein sources were casein (control group), intact CG (CG group), CG hydrolysate A (CGHA group, 30% of protein as peptides and 70% as free amino acids) and CG hydrolysate P (CGHP group, 93% of protein as peptides and 7% as free amino acids). Body weight gain, adipose tissue weights, nitrogen balance, absorptions of energy, protein and fat, lipid profiles in plasma, liver and feces and hepatic activities of carnitine palmitoyl transferase (CPT), fatty acid synthase (FAS), malic enzyme (ME) and glucose-6-phosphate dehydrogenase (G6PDH) were assessed. The CGHA diet had the highest amount of BCAAs, especially leucine, and most of them existed as free amino acid forms. The CGHA group showed significant weight reduction and negative nitrogen balance. Protein absorption and apparent protein digestibility in the CGHA group were significantly lower than those in other groups. Adipose tissue weights were the lowest in the CGHA group. Activity of CPT tended to be higher in the CGHA group than in other groups and those of FAS, ME and G6PDH were significantly lower in the CGHA group than in other groups. In conclusion, the CGHA diet which had relatively high amounts of free amino acids and BCAAs, especially leucine, had a weight reduction effect by lowering adipose tissue weight and the activities of FAS, ME and G6PDH in experimental animals, but it seemed to be a negative result induced by lowering protein absorption, increasing urinary nitrogen excretion and protein catabolism

The interfacial bioscience grand challenge.

This report is broken down into the following 3 sections: (1) Chemical Cross-linking and Mass Spectrometry Applied to Determination of Protein Structure and Dynamics; (2) Computational Modeling of Membrane Protein Structure and Dynamics; and (3) Studies of Toxin-Membrane Interactions using Single Molecule Biophysical Methods

Evaluation of Antioxidant Effects of Pumpkin (<i>Cucurbita pepo</i> L.) Seed Extract on Aging- and Menopause-Related Diseases Using Saos-2 Cells and Ovariectomized Rats

Aging and menopause are associated with oxidative stress and inflammation. Here, we evaluated the antioxidant properties of pumpkin (Cucurbita pepo L.) seed extract and assessed its ameliorative effects on aging- and menopause-related diseases using Saos-2 cells and ovariectomized rats. The seed extract had bioactive components that exhibited antioxidant activity. The extract increased the alkaline phosphatase (ALP) activity of Saos-2 cells. The oral administration of the extract to ovariectomized rats for 12 weeks decreased their body weight, fat weight, and cardiac risk indices. It also contributed to reductions in the levels of reactive oxygen species, oxidative stress, and inflammation, as assessed by measuring the serum levels of malondialdehyde and analyzing gene expression in rats. Furthermore, the administration of the extract also promoted an enhancement of the transcription of nuclear factor erythroid 2-related factor (Nrf2), heme oxygenase-1 (Ho-1), and catalase (Cat), involved in antioxidant activity; endothelial nitric oxide synthase (eNos), involved in vasculoprotective activity; and PR/SET domain 16 (Prdm16) and peroxisome proliferator-activated receptor-gamma coactivator (Pgc1α), involved in brown adipogenesis and thermogenesis. Our results using ovariectomized rats show that pumpkin seed extract may have ameliorative effects on menopause-related diseases by increasing ALP activity, evaluating the antioxidant system, ameliorating oxidative stress and thermogenesis, and enhancing lipid profiles

The Attitudes of Medical Students for Their Roles and Social Accountability in the COVID-19 Pandemic Era

Introduction In this study, we aimed to explore the attitude of medical students toward their roles and social accountability in this pandemic era. An online survey asked questions covering the topics of 1) the role of medical students in the pandemic era; 2) Medical education in the ‘new normal,’ and 3) the impact of COVID-19 on medical students. Methods The online survey, developed by a team consisting of 3 medical students, 3 psychiatric residents, and 3 psychiatric professors, was distributed to medical students, graduates, and professors in a single South Korean medical school. Anxiety symptom rating scales, including the Stress and Anxiety to Viral Epidemic - 6 (SAVE-6) scale and the Generalized Anxiety Disorder - 7 (GAD-7) scale, were also applied to measure participant anxiety level. Results All of the 213 medical students, 180 graduates, and 181 professors that participated in this online survey were at the same medical school. Medical students indicated their willingness to join the healthcare response to the COVID-19 pandemic if requested; however, graduates and professors recommended that medical students continue their medical school curriculum rather than join the response. In the new normal era, medical education was considered to be changed appropriately. Moreover, adequate knowledge of COVID-19 infection and spread must be considered for the continuation of clinical clerkships during the pandemic. Overall, medical students who indicated anxiety about treating possible or confirmed cases rated higher on the SAVE-6 scale. Finally, medical students who reported that COVID-19 had an influence on their studies and daily life rated higher on the general anxiety scale (GAD-7). Conclusion Social accountability is an important issue for medical students in the pandemic era. At the same time, cultivating professionalism is also important for the readiness for the future healthcare responses

Comparison of the FLUKA, MCNPX, and PHITS Codes in Yield Calculation of Secondary Particles Produced by Intermediate Energy Proton Beam

The accurate estimation of secondary particle production from thick targets by intermediate energy proton or heavy ion is very important factor to determine source term in radiation shielding design. Three well-proved Monte Carlo codes of the FLUKA, MCNPX, and PHITS were reviewed and benchmarking calculations for proton-induced neutrons and photons have been carried out. Neutron yields from Be, C, Al, and Fe target for 113 and 256 MeV protons were calculated and compared with Meier&apos;s experimental data. From those comparisons, the property of above Monte Carlo codes could be observed for application of shielding design. The proton energies of 100, 150, 200 and 230 MeV were reviewed for thick Al, Fe, Cu, and Pb targets to develop the source term. Dependence properties of neutron and photon production yields were found for target materials and target thickness. In this paper, a part of comparison results are presented and the discrepancies and agreements between each code are discussed for various target materials and proton energies

Immiscible Oil–Water Interface: Dual Function of Electrokinetic Concentration of Charged Molecules and Optical Detection with Interfacially Trapped Gold Nanorods

In this paper, we report that an immiscible oil–water interface can achieve the dual function of electrokinetic molecular concentration without external electric fields and sensitive optical detection without a microscope. As a proof-of-concept, we have shown that the concentration of positively charged molecules at the oleic acid–water interface can be increased significantly simply by controlling the pH. Three-dimensional phase field simulation suggests that the concentration of positively charged rhodamine 6G can be increased by about 10-fold at the interface. Surface-enhanced Raman spectroscopy (SERS) is utilized for label-free detection by taking advantage of this molecular accumulation occurring at the interface, since gold nanorods can be spontaneously trapped at the interface via electrostatic interaction. SERS measurements suggest that the immiscible oleic acid–water interface allows the limit of detection to be improved by 1–3 orders of magnitude