Resveratrol inhibits rhinovirus replication and expression of inflammatory mediators in nasal epithelia

Significance Statement

Human rhinoviruses (HRV) are responsible for at least 50% of the common colds, a mild, self-limiting upper respiratory tract illness, nonetheless with major economic impact through loss of productivity. HRV infection is also associated with acute otitis media, sinusitis, bronchiolitis, pneumonia, and severe infections, especially in immunocompromised patients and is also the major cause of exacerbations in both asthma and chronic obstructive pulmonary disease.

There are no approved antiviral agents for the prevention or treatment of HRV infection and vaccination is not feasible since more than 100 different rhinovirus types with low antibody cross-reactivity have been described.

Natural bioactive compounds could represent an attractive therapeutic tool to counteract infectious agents. Among natural polyphenols, Resveratrol is of particular interest. It is produced by several plants in response to stress or injury induced by microorganisms or environmental hazards and protects fruits and vegetables against infections. Resveratrol displays a wide range of biological and pharmacological activities including anti-inflammatory, antioxidative, anticancer, antibacterial, antiviral, cardioprotective and neuroprotective effects. Several studies have demonstrated that resveratrol exerts antiviral effect against different viruses, including herpes simplex virus, human cytomegalovirus, varicella- zoster virus, Epstein Barr virus, influenza A virus, respiratory syncytial virus, and HIV, but to date no studies demonstrated an effect of resveratrol on HRV. The study presented here provides evidence for a high, dose-dependent, antiviral activity against HRV replication in human nasal epithelia. Resveratrol also strongly suppresses HRV-induced inflammatory mediators and ICAM-I expression. Carboxymethylated glucan improves the stability of resveratrol and further increases the anti-inflammatory effect of the polyphenol. The possibility that a natural compound can be used to affect viral replication and pro-inflammatory cytokine production represents an ideal therapeutic approach. Such therapy, particularly if topical, could have obvious implications for the treatment of rhinovirus infections and the prevention of complications.

Journal Reference

Antiviral Res. 2015 Nov;123:15-21.

Mastromarino P¹, Capobianco D², Cannata F², Nardis C², Mattia E², De Leo A², Restignoli R³, Francioso A⁴, Mosca L⁴.

Show Affiliations

1. Department of Public Health and Infectious Diseases, Section of Microbiology, Sapienza University, I-00185 Rome, Italy. Electronic address: paola.mastromarino@uniroma1.it.
2. Department of Public Health and Infectious Diseases, Section of Microbiology, Sapienza University, I-00185 Rome, Italy.
3. NOOS s.r.l., I-00181 Rome, Italy.
4. Department of Biochemical Sciences, Sapienza University, I-00185 Rome, Italy.

Abstract

Human rhinoviruses (HRV), the cause of common colds, are the most frequent precipitants of acute exacerbation of asthma and chronic obstructive pulmonary disease, as well as causes of other serious respiratory diseases. No vaccine or antiviral agents are available for the prevention or treatment of HRV infection. Resveratrol exerts antiviral effect against different DNA and RNA viruses. The antiviral effect of a new resveratrol formulation containing carboxymethylated glucan was analyzed in H1HeLa cell monolayers and ex vivo nasal epithelia infected with HRV-16. Virus yield was evaluated by plaque assay and expression of viral capsid proteins by Western blot. IL-10, IFN-β, IL-6, IL-8 and RANTES levels were evaluated by ELISA assay. ICAM-1 was assessed by Western blot and immunofluorescence. Resveratrol exerted a high, dose-dependent, antiviral activity against HRV-16 replication and reduced virus-induced secretion of IL-6, IL-8 and RANTES to levels similar to that of uninfected nasalepithelia. Basal levels of IL-6 and RANTES were also significantly reduced in uninfected epithelia confirming an anti-inflammatory effect of the compound. HRV-induced expression of ICAM-1 was reversed by resveratrol. Resveratrol may be useful for a therapeutic approach to reduce HRV replication and virus-induced cytokine/chemokine production.

Copyright © 2015 Elsevier B.V. All rights reserved.

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Oct-2 forms a complex with Oct-1 on the iNOS promoter and represses transcription by interfering with recruitment of RNA PolII by Oct-1

Significance Statement

Transcriptional regulation is the fundamental process that modulates cell behaviour in response to environmental cues as well as intracellular requirements ultimately leading to the control of cellular responses in the organism. One of these processes is innate immunity, the first line defence against infectious microorganisms and neoplastic cells. A vital component of this system is the inducible nitric oxide synthase (iNOS) that produces high levels of nitric oxide (NO) in response to immune stimuli which is able to kill neighbouring cells and microbes in a non-specific manner. Due to the toxicity of these levels of NO, expression of iNOS is tightly regulated, mainly at the level of transcription. We showed previously that Oct-1 (POU2f1) is an essential transcription factor that binds to the proximal promoter and regulates iNOS, by recruiting RNA Pol II, leading to initiation and pausing. The relevance of this regulatory mechanism is that iNOS is primed for transcription so that when a stimulus triggers activation of transcription factors such as NFkB, transcription elongation can proceed swiftly to generate a rapid response.

Oct-1 is a member of the POU family of transcription factors that includes seven classes of regulators with distinct function and expression patterns. A common feature of this family is a central DNA-binding domain which in many members has affinity for the same octamer element ATGCAAAT. The flanking N- and C- terminus domains confer each member their specific activity. Oct-2 (POU2f2) is another member of this family which is closest to Oct-1 and whose expression is confined to B cells and neurones. Functionally, it has been implicated in B cell development and regulation of B lymphocyte-specific genes.

One of the many puzzles of transcriptional regulation is how different transcription factors with the same DNA binding domain can have different effects on gene expression. Though many of these differences can be explained by cell type-specific expression of these factors, this is not always clear. For example, though Oct-2 is normally restricted to B lymphocytes and neurones, we found it expressed in some cancer cell lines. We observed that in the cell lines that express Oct-2, iNOS is repressed. The mechanism of Oct-2 repression of iNOS consists in the formation of a complex with Oct-1 on the proximal promoter of the iNOS gene. The presence of Oct-2 interferes with Oct-1 activity to recruit and activate PolII thereby precluding transcription.

These findings help understand the mechanisms by which transcription factors control protein expression in a cell-specific manner and how the interplay between them achieves a fine tuning in response to particular circumstances. The wider interaction of the regulators includes the basal transcriptional machinery as well as the chromatin itself. Furthermore, though these mechanisms allow a differential regulation of proteins in specific cell types, for example avoiding the expression of the potentially harmful iNOS in sensitive cells such as neurones, they can also be hijacked by cancer cells to evade death signalling. The mechanisms that control gene expression are varied and versatile and a better understanding of the molecular basis for them will allow us to modulate them in health and disease. 

Journal Reference

Nucleic Acids Res. 2015 Nov 16;43(20):9757-65.

Bentrari F¹, Chantôme A¹, Knights A², Jeannin JF¹, Pance A³. 

Show Affiliations

1. EPHE Laboratory, Faculty of Medicine, University of Bourgogne, 7 Boulevard Jeanne D’Arc, 21033 Dijon, France.
2. The Wellcome Trust Sanger Institute, Genome Campus, Hinxton, Cambridge CB10 1SA, UK.
3. EPHE Laboratory, Faculty of Medicine, University of Bourgogne, 7 Boulevard Jeanne D’Arc, 21033 Dijon, France The Wellcome Trust Sanger Institute, Genome Campus, Hinxton, Cambridge CB10 1SA, UK ap9@sanger.ac.uk.

Abstract

Oct-1 (POU2f1) and Oct-2 (POU2f2) are members of the POU family of transcription factors. They recognize the same DNA sequence but fulfil distinct functions: Oct-1 is ubiquitous and regulates a variety of genes while Oct-2 is restricted to B-cells and neurones. Here we examine the interplay and regulatory mechanisms of these factors to control the inducible nitric oxide synthase (iNOS, NOS2). Using two breast cancer cell lines as a comparative model, we found that MCF-7 express iNOS upon cytokine stimulation while MDA-MB-231 do not. Oct-1 is present in both cell lines but MDA-MB-231also express high levels of Oct-2. Manipulation of Oct-2 expression in these cell lines demonstrates that it is directly responsible for the repression of iNOS in MDA-MB-231. In MCF-7 cells Oct-1 binds the iNOS promoter, recruits RNA PolII and triggers initiation of transcription. In MDA-MB-231 cells, both Oct-1 and Oct-2 bind the iNOS promoter, forming a higher-order complex which fails to recruit RNA PolII, and as a consequence iNOS transcription does not proceed. Unravelling the mechanisms of transcription factor activity is paramount to the understanding of gene expression patterns that determine cell behaviour.

© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

Go ToNucleic Acids Res.  

Figure Legend

BLUE: POU2f1 (Oct-1) binds to the proximal promoter, recruits RNAPolII and triggers initiation of transcription. Upon immune stimulation, NFkB is activated and triggers elongation for rapid expression of iNOS

RED: When POU2f2 (Oct-2) is present in the cell, it forms a heterodimer with Oct-1 on the proximal promoter, Interfering with the recruitment of RNAPolII and thereby repressing transcription.

A fetal human heart cardiac-inducing RNA (CIR) promotes the differentiation of stem cells into cardiomyocytes

 Summary

We have discovered a unique RNA that has the ability to transform non-muscle cells into contracting cardiac muscle tissue using cardiac non-function mutant Mexican axolotl (salamander) embryos as “tools” in the study.  In the present study, RNAs derived by molecular cloning of commercially-obtained human fetal heart RNA were tested to determine if human RNA also might have the ability to promote cardiomyocyte differentiation from non-muscle cells. The human fetal heart RNA was cloned, sequenced and synthesized using standard laboratory protocols and then tested initially using the cardiac non-function axolotl embryonic heart bioassay system. This human-derived RNA was able to rescue the non-functional mutant hearts by converting non-muscle cells in their heart walls into contracting myocardial tissue with well-organized sarcomeric myofibrils. Similar results were obtained using undifferentiated mouse embryonic and human-induced pluripotent stem cells in culture.  Both mouse- and human-derived pluripotent stem cells transfected with the Cardiac Inducing RNA ( CIR) form into shapes characteristic of early cardiac myocytes in culture and express the cardiac specific contractile protein marker, troponin-T, in addition to tropomyosin and α-actinin as detected by immunohistochemical staining. Expression of these contractile proteins also shows organization into sarcomeric myofibrils characteristic of striated cardiac muscle cells. The active clone producing the cardiac-inducing RNA (CIR) was found to be a fragment of the N-sulfoglucosaminesulfohydrolase and the caspase recruitment domain family member 14 percursor.   Computer analyses of the RNA secondary structures of this active CIR using the Genebee Computer Structural Analysis Program developed at the Belozersky Research Institute of Moscow State University, Russia, show significant similarities to the Myocardial Cell Inducing RNA (MIR) from axolotl which also promotes non-muscle cells to differentiate into cardiac muscle. Thus, these two RNAs, salamander MIR and the newly-discovered human-derived Cardiac Inducing RNA reported here, appear to have evolutionarily conserved secondary structures suggesting that both play major roles in vertebrate heart development and, particularly, in the differentiation of cardiomyoctyes from non-muscle cells.

Significance Statement

Of the hundreds of thousands of people who suffer heart attacks each year, in spite of current medical treatments that improve heart function and quality of life for some, many do not significantly recover because connective scar tissue, rather than new muscle, replaces the cardiac muscle in the infarcted area. The research reported here may very well lead to better treatments in the future for patients recovering from myocardial infarctions or other heart diseases that adversely affect the myocardial muscle tissue. Being able to use the Cardiac Inducing RNA (CIR) itself, or in combination with induced pluripotent stem cells derived from that same patient to repair the damaged heart muscle tissue or replace the scar tissue with vigorously contracting normal myocardial cells, could lead to a much better prognosis for  cardiac patients and may very well allow them to completely recover and return to pre-heart-attack activity levels.

 

About The Author

Larry F. Lemanski, Ph.D.

Larry F. Lemanski completed his B.S. degree with honors, in Biology and Chemistry, from the University of Wisconsin-Platteville and his M.S. and Ph.D. degrees from Arizona State University, Tempe, Arizona, U.S.A.  After four years as an NIH and MDAA Postdoctoral fellow at the University of Pennsylvania, Philadelphia, he joined the medical faculty at the University of California, San Francisco, as an Assistant Professor in residence and then moved to the faculty of the College of Medicine at the University of Wisconsin, Madison, where he went up through the ranks to Full Professor of Anatomy. He then joined the Upstate Medical University in Syracuse, New York, as Professor and Chairman of the Department of Anatomy and Cell Biology. He moved to Texas A&M University as an Associate Vice President for Research, then to Florida Atlantic University as a Vice President for Research, to Temple University as Senior Vice president for Research and Strategic Initiatives, and to Texas A&M University-Commerce as Provost of the University. He has now transitioned to the position of Distinguished Research Professor at Texas A&M University-Commerce. His research interests throughout his career have involved embryonic heart development and cardiac cell differentiation using a variety of morphological, cellular, biochemical, and molecular biology approaches. He and his associates have published numerous papers in the field of embryonic heart development and cardiac cell differentiation. His current work centers around his earlier pioneering discovery that selected RNAs can promote the differentiation of non-muscle cells to form into functional cardiac tissue.

About The Author

Ashley Moses-Arms, B.S., M.S.

Ashley Moses-Arms completed her B.S degree with highest honors (summa cum laude) from Texas A&M University-Commerce Honors College majoring in the Biological Sciences. Her honors thesis research was done under the mentorship of Professor Larry Lemanski and involved studies on a human-derived RNA that promotes cardiac muscle differentiation in cardiac mutant non-function salamanders. She completed a M.S. degree in the Biological Sciences Program also from Texas A&M University-Commerce in Dr. Venugopalan Cheriyath’s laboratory. Her interests have been in the areas of regenerative medicine and cancer research. Currently she is a first year medical student in the College of Medicine at the University of Texas Medical Branch in Galveston.

 

About The Author

Andrei Kochegarov, Ph.D.

Dr. Andrei Kochegarov completed his undergraduate and Master’s degrees in Biology at Saratov State University, Russia and his Ph.D. in Molecular and Cellular Biology at Pushchino State University, Russia. He worked as a Postdoctoral Research Associate for ten years at major research Universities in California, and the last five years, he has been a Research Assistant Professor working on stem cell research and cardiac regeneration at Texas A&M University, Commerce, as well as a lecturer for several Biology courses. He is interested in Regenerative Biology, Cardiac and Neuronal Regeneration, Oxidative Stress, Cellular Aging and Cancer Research.

Journal Reference

In Vitro Cell Dev Biol Anim. 2015; 51(7):739-48.

Kochegarov A, Moses-Arms A, Lemanski LF.

Department of Biological and Environmental Sciences, Texas A&M University-Commerce, Commerce, TX, 75429-3011, USA.

Abstract

A specific human fetal heart RNA has been discovered, which has the ability to induce myocardial cell formation from mouse embryonic and human-induced pluripotent stem cells in culture. In this study, commercially obtained RNA from human fetal heart was cloned, sequenced, and synthesized using standard laboratory approaches. Molecular analyses of the specific fetal cardiac-inducing RNA (CIR), revealed that it is a fragment of N-sulfoglucosaminesulfohydrolase and the caspase recruitment domain family member 14 precursor. Stem cells transfected with Cardiac Inducing RNA often form into spindle-shaped cells characteristic of cardiomyocytes,and express the cardiac-specific contractile protein marker, troponin-T, in addition to tropomyosin and α-actinin as detected by immunohistochemical staining. Expression of these contractile proteins showed organization into sarcomeric myofibrils characteristic of striated cardiac muscle cells. Computer analyses of the RNA secondary structures of the active Cardiac Inducing RNA show significant similarities to a RNA from salamander or myofibril-inducing RNA (MIR), which also promotes non-muscle cells to differentiate into cardiac muscle. Thus, these two RNAs, salamander MIR and the newly discovered human-cloned cardiac inducing RNA reported here, appear to have evolutionarily conserved secondary structures suggesting that both play major roles in vertebrate heart development and, particularly, in the differentiation of cardiomyocytes from non-muscle cells during development.

Go To In Vitro Cell Dev Biol Anim. Acknowledgments:  This work is supported by NIH grant (HL061246), NSF grant (1121151), and an American Heart Association grant (10GRNT4530001) awarded to LFL.

Figure Legends

Figure 1. Mouse embryonic stem cells transfected with Cardiac Inducing RNA (CIR), cultured for 8 days and immunostained with antibody against cardiac-specific troponin-T.  These CIR-treated cells show significant staining for cardiac troponin-T, indicating they have differentiated into cardiac lineages.  The DAPI-stained blue nuclei are difficult to visualize in these cells due to the heavy FITC-staining for cardiac specific troponin-T.

Figure 2. Mouse embryonic stem cell controls which have been cultured for 8 days, stained with antibodies against cardiac-specific troponin-T but not transfected with the cardiac Inducing RNA (CIR).  Without the CIR treatment, these cells do not express significant amounts of cardiac troponin-T.  Mostly, only the DAPI –stained blue nuclei are visible.

Figure 3. High magnification fluorescent micrograph of a human induced pluripotent stem cell (iPSC) treated with Cardiac Inducing RNA (CIR), cultured for 8 days and stained with cardiac-specific anti-troponin-T antibodies.  This spindle-shaped cell, shows staining for cardiac-specific troponin-T which is organized into myofibrillar- like structures characteristic of early developing cardiomyocytes.   (From:  Kochegarov A., Moses-Arms, A., Lemanski, L.F.  2015 A fetal human heart cardiac inducing RNA (CIR)  promotes the differentiation of stem cells into cardiomyocytes.  In Vitro Cell Dev Biol-Animal  DOI 10.1007/s11626-015-9880-4.  With permission of the authors).

 

 

 

 

 

Low-Density Lipoprotein Apheresis Ameliorates Renal Prognosis of Cholesterol Crystal Embolism

Journal Reference

Ther Apher Dial. 2015;19(4):355-60.

Ishiyama K, Sato T, Taguma Y. 

Department of Nephrology, Japan Community Health Care Organization Sendai Hospital, Sendai, Japan.

Abstract

Drugs such as corticosteroids and statins have been used to treat cholesterol crystal embolism (CCE), but the prognosis remains poor. This study evaluated the efficacy of low-density lipoprotein apheresis (LDL-A) in patients with cholesterol crystal embolism. Patients with cholesterol crystal embolism who showed renal deterioration after vascular interventions were studied retrospectively. Information on demographic variables, clinical measurements, and medication use was collected. The outcomes were incidence of maintenance dialysis and mortality at 24 weeks. A total of 49 patients with cholesterol crystal embolism were included, among whom 37 (76%) were diagnosed pathologically and the remainder  were diagnosed clinically. The median estimated GFR at baseline and at diagnosis were 40.5 and 13.4 mL/min per 1.73 m(2) , respectively. Corticosteroids were used in 42 patients (86%), statins in 30 patients (61%), and angiotensin-converting enzyme inhibitors and angiotensin receptor blockers in 29 patients (59%). LDL-A was performed in 25 patients (LDL-A group), and not in 24 patients (control group). Smoking (100% vs. 72%, P = 0.02), white blood cell count (8900/mm(3) vs. 7000/mm(3) ) and corticosteroid use (96% vs. 75%) were higher in the LDL-A group compared with the control group, but there were no differences in other demographic and clinical parameters between the groups. Patients in the LDL-A group had a lower incidence of maintenance dialysis (2/25 (8%) vs. 8/24 (33%), P < 0.05), and a trend towards lower mortality (2/25 (8%) vs. 7/24 (29%), P = 0.074). These results suggest that LDL-A decreases the risk of maintenance dialysis in severe renal cholesterol crystal embolism patients after vascular interventions.

© 2015 International Society for Apheresis, Japanese Society for Apheresis, and Japanese Society for Dialysis Therapy.

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Figure: the picture shows typical findings of renal cholesterol crystal embolism in biopsy specimen.

 

Quantitative analysis of cadherin-11 and β-catenin signalling during proliferation of rheumatoid arthritis-derived synovial fibroblast cells

Journal Reference

J Pharm Pharmacol. 2015;67(8):1075-82.

Yoshioka R¹, Kita Y¹, Nagahira A², Manno A¹, Makita N¹, Tomita U¹, Murakawa M¹.

Show Affiliations

1. Faculty of Exploratory Pharmacology, Asubio Pharma Co., Ltd., Kobe, Japan.
2. Faculty of Drug Discovery Technology, Asubio Pharma Co., Ltd., Kobe, Japan.

Abstract

OBJECTIVES:

Cadherin-11 (CDH11) is an adhesion molecule that anchors β-catenin and is involved with various functions of synovial fibroblast cells (SFCs) during the development of rheumatoid arthritis (RA). However, the mechanism of Cadherin-11 during RA-SFC proliferation is unclear. The aim of our study was to clarify the involvement of Cadherin-11 and β-catenin signalling during proliferation.

METHODS:

IL-1β-induced and tumour necrosis factor-α (TNF-α)-induced cell proliferation, with Cadherin-11 siRNAs, β-catenin-specific siRNAs and a Cadherin-11-neutralizing antibody, were assessed by 5-Bromo-2′-deoxy-uridine ELISA.

KEY FINDINGS:

Using Cadherin-11 siRNAs, there were a 42% reduction in IL-1β-induced proliferation and a 64% reduction in β-catenin protein. When β-catenin siRNAs were applied, there was a 63% reduction in IL-1β-induced proliferation. The median effective concentration (EC50 ) values for IL-1β-induced proliferation via CDH11-mediated β-catenin-dependent, total β-catenin-dependent and β-catenin-independent signalling were 0.0015, 0.016 and 0.18 ng/ml, respectively. Blocking CDH11 ligation with a CDH11-neutralizing antibody did not decrease IL-1β-induced proliferation.

CONCLUSIONS:

CDH11-mediated β-catenin signalling was 42% involved in IL-1β-induced proliferation and had the highest susceptibility to IL-1β among the proliferative signallings analysed in this study. The mode of action for Cadherin-11 during the cell proliferation was likely associated with a pool of β-catenin protein. In contrast, Cadherin-11 and β-catenin were not involved in TNF-α-induced RA-SFC proliferation.

© 2015 Royal Pharmaceutical Society.

Go To J Pharm Pharmacol.

 

Conjugated Polythiophene for Rapid, Simple, and High-Throughput Screening of Antimicrobial Photosensitizers

Significance Statement

  Photodynamic antimicrobial chemotherapy (PACT) is effective in killing microbial cells with resistance to antibiotics and can be designed to target localized infections and offers little possibility to produce photoresistant species. Since the behavior of photosensitizers (PS) is crucial for enhancing the efficiency of PACT, accessing and finding suitable and efficient PS are becoming extremely important. Researchers from Hebei University of Technology, P.R. China reported a new, rapid, simple and high-throughput method with only a single cationic conjugated polythiophene derivative (PMNT) to realize the screening of photosensitizer activity. This new strategy has three significant characteristics. First, the PMNT can be simply used to quantitatively measure the amounts of bacteria in high sensitivity. Second, no expensive materials and instruments were required, leading to the low cost of our method (~$1.0 for one high-throughput screening of six photosensitizers ). Third, the high-throughput screening of PACT efficacy only takes 4.5 hours to complete the assay including incubation, irradiation, culture and detection. This new platform is efficient and promising for discovering new kinds of PACT photosensitizers. 

About The Author

Chengfen Xing is a full professor of chemistry at School of Sciences, Hebei University of Technology. She earned her Ph.D. in Chemistry from the Institute of Chemistry, Chinese Academy of Sciences in 2011. She was a postdoctoral researcher at MESA⁺ Institute of Nanotechnology, University of Twente and was a postdoctoral researcher at Institute of Molecules and Materials, University of Nijmegen from 2011 to 2012. From 2013, she began her independent faculty career as a full professor of Chemistry, Hebei University of Technology. Her current research interest is focused on design, synthesis of conjugated polymers-based hybrid materials for biosensing and building stimuli-responsive architectures.

Journal Reference

ACS Appl Mater Interfaces. 2015 Jul 15;7(27):14569-72.

Li R¹, Niu R², Qi J², Yuan H², Fan Y², An H², Yan W¹, Li H¹, Zhan Y², Xing C^1,2. 

Show Affiliations

1. School of Chemical Engineering and Technology, Hebei University of Technology, Tianjin 300130, P. R. China.
2. Key Laboratory of Hebei Province for Molecular Biophysics, Institute of Biophysics, Hebei University of Technology, Tianjin 300401, P. R. China.

Abstract

The cationic conjugated poly[3-(3′-N,N,N-triethylamino-1′-propyloxy)-4-methyl-2,5-thiophene hydrochloride] (PMNT) has been developed for high-throughput screening of photodynamic antimicrobial  chemotherapy photosensitizers (PSs). The bacterial number can be detected quantitatively by PMNT via various fluorescence quenching efficiencies. The photosensitized inactivation  of bacteria is not efficient with ineffective PSs, and thus the bacteria grow exponentially and can   be coated tightly by PMNT through electrostatic and hydrophobic interactions, resulting in aggregates  and fluorescence quenching of PMNT, whereas, conversely, effective photosensitizers lead to original and strong fluorescence of PMNT. This new platform of high-throughput screening is promising for discovering  new photosensitizers.

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Noncationic Rigid and Anisotropic Coiled-Coil Proteins Exhibit Cell-Penetration Activity

Significance Statement

The use of cationic polymers and cationic cell-penetrating peptides as non-viral carriers in cellular delivery systems has been studied, but polycations are usually faced with the problems such as inflammatory nature and cytotoxicity. Researchers in Japan, Nippon Institute of Technology, RIKEN, and Hokkaido University have previously reported that artificial proteins having rigid and anisotropic structure and surface cationic property showed superior cell-penetrating activities. In this report, the authors designed novel artificial protein variants of rigid and anisotropic structure but noncationic surface to avoid problems related to inflammation and cytotoxicity. These artificial proteins showed less cell-penetrating activity than previous cationic artificial protein, however, they were able to penetrate cells comparable to cationic cell-penetrating peptides. Further, one variant did not show short-term cytotoxicity in all the cells tested. These findings provide new insights for the design of non-viral and non-cationic carriers in cellular delivery systems with excellent efficacy and safety.

About The Author

Ken-Ichi Sano is an associate professor of Innovative Systems Engineering at Nippon Institute of Technology. He received his PhD from Nagoya University and was a postdoctoral fellow at RIKEN in SPring-8, Cancer Institute Japanese Foundation for Cancer Research, and was a Deputy Unit Leader at RIKEN in Wako. He was the recipient of the Lewis Dienes Award of the 10th International Organization of Mycoplasmology, and the Young Investigator Award of the 4th International Peptide Meeting. His current research interests include cellular drug delivery systems, evaluation of antidepressant agents using novel model systems, and creation of functional artificial proteins.

Journal Reference

Langmuir. 2015;31(30):8218-23.

Nakayama N¹, Hagiwara K^1,2, Ito Y¹, Ijiro K^1,3, Osada Y¹, Sano K¹. 

Show Affiliations

1. Nano Medical Engineering Laboratory, RIKEN, Wako, Saitama 351-0198, Japan.
2. Tokyo Metropolitan Institute of Medical Science, Setagaya, Tokyo 156-8506, Japan.
3. Research Institute for Electronic Science, Hokkaido University, Sapporo, Hokkaido 001-0021, Japan

.

Abstract

Numerous cationic peptides that penetrate cells have been studied intensively as drug delivery system carriers for cellular delivery. However, cationic molecules tend to be cytotoxic and cause inflammation, and their stability in the blood is usually low. We have previously demonstrated that a rigid and fibrous cationic coiled-coil protein exhibited cell-penetrating ability superior to that of previously reported cell-penetrating peptides. Making use of structural properties, here we describe the cell-penetrating activity of a rigid and fibrous coiled-coil protein with a noncationic surface. A fibrous coiled-coil protein of pI 6.5 penetrated 100% of the cells tested in vitro at a concentration of 500 nM, which is comparable to that of previously reported cell-penetrating peptides. We also investigated the effect of cell-strain dependency and short-term cytotoxicity.

Go To Langmuir.

Figure Legend: Summary of cell-penetrating activity of artificial protein having rigid and anisotropic structure with various surface properties.

 

 

 

 

 

Application of clotrimazole via a novel controlled release device provides potent retinal protection

Journal Reference

J Mater Sci Mater Med. 2015; 26(9):230.

Nezhad ZK, Nagai N, Yamamoto K, Kaji H, Nishizawa M, Saya H, Nakazawa T, Abe T.

Division of Clinical Cell Therapy, United Centers for Advanced Research and Translational Medicine (ART), Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, 980-8575, Japan.

Abstract

Age-related macular degeneration is the leading cause of legal blindness among older individuals. Therefore, the development of new therapeutic agents and optimum drug delivery systems for its treatment are crucial. In this study, we investigate whether clotrimazole (CLT) is capable of protecting retinal cells against oxidative-induced injury and the possible inhibitory effect of a sustained clotrimazole-release device against light-induced retinal damage in rats. In vitro results indicated pretreatment of immortalized retinal pigment epithelium cells (RPE-J cells) with 10-50 µM clotrimazole before exposure to oxygen/glucose deprivation conditions for 48 h decreased the extent of cell death, attenuated the percentage of reactive oxygen species-positive cells, and decreased the levels of cleaved caspase-3. The device consists of a separately fabricated reservoir, a clotrimazole formulation, and a controlled  release cover, which are made of poly(ethyleneglycol) dimethacrylate (PEGDM) and tri(ethyleneglycol) dimethacrylate (TEGDM). The release rate of clotrimazole was successfully tuned by changing the ratio of PEGDM/TEGDM in the cover. In vivo results showed that use of a CLT-loaded device lessened the reduction of electroretinographic amplitudes after light exposure. These findings indicate that the application of a polymeric clotrimazole-loaded device may be a promising method for the treatment of some retinal disorders.

Go To J Mater Sci Mater Med.

 

 

 

 

 

Crocetinic acid inhibits hedgehog signaling to inhibit pancreatic cancer stem cells

Journal Reference

Oncotarget. 2015;6(29):27661-73.

Rangarajan P¹, Subramaniam D¹, Paul S², Kwatra D¹, Palaniyandi K², Islam S², Harihar S², Ramalingam S¹, Gutheil W³, Putty S³, Pradhan R⁴, Padhye S⁴,Welch DR^2,1, Anant S^2,1, Dhar A^2,1.

Show Affiliations

1. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS, USA.
2. Department of Cancer Biology, University of Kansas Medical Center, Kansas City, KS, USA.
3. Department of Pharmaceutical Sciences, University of Missouri at Kansas City, Kansas City, MO, USA.
4. Interdisciplinary Science and Technology Research Academy, Abeda Inamdar College, University of Pune, Pune, India.

Abstract

Pancreatic cancer is the fourth leading cause of cancer deaths in the US and no significant treatment is currently available. Here, we describe the effect of crocetinic acid, which we purified from commercial saffron compound crocetin using high performance liquid chromatography. Crocetinic acid  inhibits proliferation of pancreatic cancer cell lines in a dose- and time-dependent manner. In addition, it induced apoptosis. Moreover, the compound significantly inhibited epidermal growth factor receptor and Akt phosphorylation. Furthermore, crocetinic acid decreased the number and   size of the pancospheres in a dose-dependent manner, and suppressed the expression of the marker protein DCLK-1 (Doublecortin Calcium/Calmodulin-Dependent Kinase-1) suggesting that crocetinic acid  targets cancer stem cells (CSC). To understand the mechanism of CSC inhibition, the signaling pathways affected by purified crocetinic acid were dissected. Sonic hedgehog (Shh) upon binding to its cognate receptor patched, allows smoothened to accumulate and activate Gli transcription factor. Crocetinic acid inhibited the expression of both Shh and smoothened. Finally, these data were confirmed in vivo where the compound at a dose of 0.5 mg/Kg bw suppressed growth of tumor xenografts. Collectively, these data suggest that purified crocetinic acid inhibits pancreatic CSC, thereby inhibiting pancreatic tumorigenesis.

=4871&pubmed-linkout=1″ target=”blank” ]Go To Oncotarget

 

Differential Expression of Sonic Hedgehog Protein in Human Hepatocellular Carcinoma and Intrahepatic Cholangiocarcinoma

Journal Reference

Pathol Oncol Res. 2015;21(4):901-8.

Al-Bahrani R1, Nagamori S, Leng R, Petryk A, Sergi C.

Department of Laboratory Medicine and Pathology, University of Alberta, 8440-112 Street, Edmonton, T6G 2B7, AB, Canada.

Abstract

Hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (CCA) are the two most common primary liver malignancies in adult patients. The molecular mechanisms underlying the pathogenesis of HCC and CCA are still poorly understood. Sonic hedgehog (SHH) signaling plays an essential role during mammalian development, i.e., promoting organ growth, tissue differentiation, and cell polarity. The upregulation of sonic hedgehog has been observed during carcinogenesis, including colorectal carcinoma. Our aim was to investigate the expression pattern of Sonic hedgehog in HCC and CCA. We investigated 40 malignant tumors of the liver, including 21 HCC and 19 of intrahepatic CCA cases by immunohistochemistry (IHC) using a polyclonal antibody against sonic hedgehog and Avidin-Biotin Complex method. We also investigated the co-localization of sonic hedgehog and Bone morphogenetic protein 4 (BMP4) in CCA using indirect double IHC. Moreover, we examined whether sonic hedgehog is expressed in two HCC cell lines HepG2 and HuH-7 and three CCA cell lines OZ, HuCCT1 and HuH28. We found that sonic hedgehog was expressed in 15 out of 21 cases (71.4 %) of HCC and 100 % of CCA cases by immunohistochemistry. Sonic hedgehog expression showed a positive trend in liver tumors (HCC, CCA) with high grade (G2-G3). Sonic hedgehog localized to the epithelial cells, while BMP4 was expressed in the stromal cells in CCA by double IHC. However, both HCC and CCA cell lines showed Sonic hedgehog expression by Western blot analysis. In conclusion, sonic hedgehog seems to be an interesting marker of de-differentiation  in liver tumors and the simultaneous epithelial-mesenchymal expression may be an intriguing prompt to investigate cross-talks between sonic hedgehog and BMP4.

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PPM1G Binds 7SK RNA and Hexim1 To Block P-TEFb Assembly into the 7SK snRNP and Sustain Transcription Elongation

Significance Statement

The transition from transcription initiation into productive elongation is a rate-limiting, highly regulated step essential for gene activation. One key regulatory factor in this transition is the positive transcription elongation factor b (P-TEFb), which phosphorylates RNA Polymerase II and negative elongation factors at promoters to relieve the block in elongation. The role of P-TEFb in activating transcription elongation programs is undisputed, yet the mechanisms through which it aintains the active transcriptional state are poorly understood. Recent work by the D’Orso lab described a detailed molecular mechanism in which a novel transcriptional co-activator, the PPM1G phosphatase, regulates the assembly of a non-coding ribonucleoprotein complex (referred to as 7SK snRNP), which recruits P-TEFb and inactivates its kinase function. Although mechanisms of P-TEFb release from the snRNP are becoming clearer, how P-TEFb remains in the 7SK-unbound state to sustain transcription elongation programs remains unknown. In response to DNA damage signaling, nuclear factor (NF)-kB rapidly recruits PPM1G to its target genes where it dephosphorylates the P-TEFb kinase thereby promoting its release from the inactive 7SK snRNP complex and promoting transcription elongation. Subsequently, PPM1G binds 7SK RNA and the P-TEFb kinase inhibitor Hexim1 to prevent reassembly of P-TEFb onto the snRNP and sustain transcription. Once the damage is resolved, PPM1G is dislodged from the snRNP and P-TEFb recycled back to form an inactive kinase complex with the 7SK snRNP at the promoter thus preventing further transcription elongation. This study provides an unprecedented example of an enzyme that regulates transcriptional activation and maintenance in response to DNA damage signaling by formation of a protein-RNA complex. 

About The Author

Dr. Aravind Gudipaty obtained her Ph.D. at the Department of Biochemistry at the University of Arizona in 2012. She then joined the laboratory of Dr. Iván D’Orso at the Department of Microbiology at the UT Southwestern Medical Center in Dallas, TX as a postdoctoral scholar from 2012-2014. In the lab of Dr. D’Orso she focused on the molecular mechanisms that control transcription elongation in response to DNA damage signaling in human cells. She is currently a postdoctoral fellow at the Huntsman Cancer Institute at the University of Utah. Her research focuses on molecular mechanisms of extrusion, which controls epithelial cell numbers during homeostasis, and how its misregulation can lead to a slew of diseases, from asthma to cancer.  

 

About The Author

Dr. D’Orso received his B.S. and Master in Biological Sciences, from the National University of Mar del Plata (Buenos Aires, Argentina) in 1998. Dr. D’Orso then obtained his Ph.D. in Biochemistry and Molecular Biology from the National University of Buenos Aires (Argentina) in 2003 under the tutelage of the U.S. National Academy Member Dr. Alberto C. Frasch. He continued his training as a post-doctoral fellow with Dr. Alan D. Frankel at the Department of Biochemistry and Biophysics at the University of California, San Francisco, during which time he was supported by fellowships from the Fundación Antorchas (Argentina), Human Frontier Science Program and amfAR. During his postdoctoral position, Dr. D’Orso elucidated key features about the mechanisms of HIV transcriptional regulation. Following receipt of a NIH Pathway to Independence Award (K99/R00) from the NIAID in 2009, Dr. D’Orso began his independent career in November 2010 at the Department of Microbiology at the UT Southwestern Medical Center in Dallas, TX. The major theme of his lab is to characterize the molecular basis of transcriptional regulation in eukaryotic cells in normal and disease states (including cancer). Dr. D’Orso’s research program is currently supported by grants from the National Institutes of Health, The Welch Foundation and an Institutional grant from the American Cancer Society.

Journal Reference

Mol Cell Biol. 2015 Nov 15;35(22):3810-28.

Gudipaty SA¹, McNamara RP¹, Morton EL¹, D’Orso I². 

Show Affiliations

1. Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
2. Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, USA Ivan.Dorso@utsouthwestern.edu.

Abstract

Transcription elongation programs are vital for the precise regulation of several biological processes. One key regulator of such programs is the P-TEFb kinase, which phosphorylates RNA polymerase II (Pol II) once released from the inhibitory 7SK small nuclear ribonucleoprotein (snRNP) complex. Although mechanisms of P-TEFb release from the snRNP are becoming clearer, how P-TEFb remains in the 7SK-unbound state to sustain transcription elongation programs remains unknown. Here we report that the PPM1G phosphatase (inducibly recruited by nuclear factor κB [NF-κB] to target promoters) directly binds 7SK RNA and the kinase inhibitor Hexim1 once P-TEFb has been released from the 7SK snRNP. This dual binding activity of PPM1G blocks P-TEFb reassembly onto the snRNP to sustain NF-κB-mediated Pol II transcription in response to DNA damage. Notably, the PPM1G-7SK RNA  interaction is direct, kinetically follows the recruitment of PPM1G to promoters to activate NF-κB transcription, and is reversible, since the complex disassembles before resolution of the program. Strikingly, we found that the ataxia telangiectasia mutated (ATM) kinase regulates the interaction between PPM1G and the 7SK snRNP through site-specific PPM1G phosphorylation. The precise and temporally regulated interaction of a cellular enzyme and a noncoding RNA provides a new paradigm for simultaneously controlling the activation and maintenance of inducible transcription  elongation programs.

Copyright © 2015, American Society for Microbiology. All Rights Reserved.

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Estrogen mimics induce genes encoding chemical efflux proteins in gram-negative bacteria

Journal Reference

Chemosphere. 2015;128:327-31.

Li X¹, Teske S¹, Conroy-Ben O².

Show Affiliations

1. Department of Civil and Environmental Engineering, 110 S. Central Campus Dr., Room 2000, Salt Lake City, UT 84112, United States.
2. Department of Civil and Environmental Engineering, 110 S. Central Campus Dr., Room 2000, Salt Lake City, UT 84112, United States. Electronic address: Otakuye.conroy@utah.edu.

Abstract

Escherichia coli and Pseudomonas aeruginosa are gram-negative bacteria found in wastewater and biosolids. Spanning the inner and outer membrane are resistance-nodulation-cell division superfamily (RND) efflux pumps responsible for detoxification of the cell, typically in response to antibiotics and other toxicity inducing substrates. Here, we show that estrogenic endocrine disruptors, common wastewater pollutants, induce genes encoding  chemical efflux proteins. Bacteria were exposed to environmental concentrations of the synthetic estrogen 17α-ethynylestradiol, the surfactant nonylphenol, and the plasticizer bisphenol-A, and analyzed for RND gene expression via q-PCR. Results showed that the genes acrB and yhiV were over-expressed in response to the three chemicals in E. coli, and support previous findings that these two transporters export hormones. P. aeruginosa contains 12 RND  efflux pumps, which were differentially expressed in response to the three chemicals: 17α-ethynylestradiol, bisphenol-A, and nonylphenol up-regulated mexD and mexF, while nonylphenol and bisphenol-A positively affected transcription of mexK, mexW, and triC. Gene expression via q-PCR of RND genes may be used to predict the interaction of estrogen mimics with RND genes. One bacterial response to estrogen mimic exposure is to induce gene expression of chemical efflux proteins, which leads to the expulsion of the contaminant from the cell.

Copyright © 2015 Elsevier Ltd. All rights reserved.

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Prevention of allergic rhinitis by ginger and the molecular basis of immunosuppression by 6-gingerol through T cell inactivation

Journal Reference

J Nutr Biochem. 2015. pii: S0955-2863(15)00226-0.

Kawamoto Y¹, Ueno Y², Nakahashi E³, Obayashi M³, Sugihara K³, Qiao S³, Iida M⁴, Kumasaka MY⁴, Yajima I⁴, Goto Y⁵, Ohgami N⁴, Kato M⁴, Takeda K³. 

Show Affiliations

1. College of Life and Health Sciences, Chubu University, Kasugai, Aichi, Japan. Electronic address: ykawa@isc.chubu.ac.jp.
2. Department of Health and Nutrition, Faculty of Psychological and Physical Science, Aichi Gakuin University, Nisshin, Aichi, Japan.
3. College of Life and Health Sciences, Chubu University, Kasugai, Aichi, Japan.
4. Department of Occupational and Environmental Health, Nagoya University Graduate School of Medicine, Nagoya, Japan.
5. Department of Biology, Faculty of Science, Toho University, Funabashi, Chiba, Japan.

Abstract

The incidence of allergies has recently been increasing worldwide. Immunoglobulin E (IgE)-mediated hypersensitivity is central to the pathogenesis of asthma, hay fever and other allergic diseases.  Ginger (Zingiber officinale Roscoe) and its extracts have been valued for their medical properties including antinausea, antiinflammation, antipyresis and analgesia properties. In this study, we investigated the antiallergic effects of ginger and 6-gingerol, a major compound of ginger, using a mouse allergy model and primary/cell line culture system. In mice with ovalbumin (OVA)-induced allergic rhinitis, oral administration of 2% ginger diet reduced the severity of sneezing and nasal rubbing by nasal sensitization of OVA and suppressed infiltration of mast cells in nasal mucosa and secretion of OVA-specific IgE in serum. 6-Gingerol inhibited the expression of not only Th2 cytokines but also Th1 cytokines in OVA-sensitized spleen cells. Accordingly, 6-gingerol suppressed in vitro differentiation of both Th1 cells and Th2 cells from naïve T cells. In addition, 6-gingerol suppressed both superantigen staphylococcal enterotoxin B (SEB)- and anti-CD3-induced T cell proliferation. 6-Gingerol also abrogated PMA plus ionomycin- and SEB-induced IL-2 production in T cells, suggesting that 6-gingerol affected T cell receptor-mediated signal transduction rather than the antigen-presentation process. Indeed, 6-gingerol inhibited the phosphorylation of MAP kinases, calcium release and nuclear localization of c-fos and NF-κB by PMA and ionomycin stimulation. Thus, our results demonstrate that 6-gingerol suppresses cytokine production for T cell activation and proliferation, thereby not causing B cell and mast cell activation and resulting in prevent ionor alleviation of allergic rhinitis symptoms.

Copyright © 2015 Elsevier Inc. All rights reserved.

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Designing Synthetic Microcapsules That Undergo Biomimetic Communication and Autonomous Motion

Journal Reference

Langmuir. 2015, 31 (44), pp 11951–11963

Yashin VV¹, Kolmakov GV², Shum H1, Balazs AC¹.

Show Affiliations

1. Chemical Engineering Department, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, United States.
2. Physics Department, New York City College of Technology, Brooklyn, New York 11201, United States.

Abstract

Inspired by the collective behavior of slime molds and amoebas, we designed synthetic cell-like objects that move and self-organize in response to self-generated chemical gradients, thereby exhibiting autochemotaxis. Using computational modeling, we specifically focused on microcapsules that encompass a permeable shell and are localized on an adhesive surface in solution. Lacking any internal machinery, these spherical, fluid-filled shells might resemble the earliest protocells. Our  microcapsules do, however, encase particles that can diffuse through the outer shell and into the surrounding fluid. The released particles play two important, physically realizable roles: (1) they affect the permeability of neighboring capsules and (2) they generate adhesion gradients on the underlying surface. Due to feedback mechanisms provided by the released particles, the self-generated adhesion gradients, and hydrodynamic interactions, the capsules undergo collective, self-sustained motion and even exhibit antlike tracking behavior. With the introduction of a chemically patterned stripe on the surface, a triad of capsules can be driven to pick up four-capsule cargo, transport this cargo, and drop off this payload at a designated site. We also modeled a system where the released particles give rise to a particular cycle of negative feedback loops (mimicking the “repressilator” network), which regulates the production of chemicals within the capsules and hence their release into the solution. By altering the system parameters, three capsules could be controllably driven to self-organize into a stable, close-packed triad that would either translate as a group or remain stationary. Moreover, the stationary triads could be made to switch off after assembly and thus produce minimal quantities of chemicals. Taken together, our models allow us to design a rich variety of self-propelled structures that achieve complex, cooperative behavior through fundamental physicochemical phenomena. The studies can also shed light on factors that enable individual protocells to communicate and self-assemble into larger communities.

Go To Langmuir.

Figure credit: Langmuir 2015.

 

 

Quantitative O-Glycomics by Microwave-Assisted β-Elimination in the Presence of Pyrazolone Analogues

Significance Statement

Quantification of O-glycomics found on the cell surface is important in physiology and pathology. Quantification will aid in understanding role of glycans in bacterial and viral recognition, cellular signaling pathways, innate immunity, cancer development. Additionally there are many glycan-specific diseases which . This study describe an improved method in quantification of o-glycomics using pyrazolone analogues.

Journal Reference

Anal Chem. 2015;87(15):7524-8.

Furukawa J¹, Piao J¹, Yoshida Y, Okada K¹, Yokota I¹, Higashino K, Sakairi N², Shinohara Y¹. 

Show Affiliations

1. Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan.
2. Graduate School of Environmental Science, Hokkaido University, Sapporo 060-0810, Japan.

Abstract

O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. The characterization of differently structured O-linked glycans (O-glycans) is particularly challenging because there is no known endoglycosidase for such groups. Therefore, chemical digestion approaches have been widely used; however, it is sometimes difficult to suppress unwanted side reactions. Recently, we reported a novel O-glycomics procedure using β-elimination in the presence of pyrazolone  analogues (BEP). In the present study, we describe a microwave (MW)-assisted pyrazolone analogues  procedure for rapid and quantitative O-glycomic analysis. Following optimization of the reaction conditions, the MW-assisted pyrazolone analogues  reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted pyrazolone analogues  procedure enabled O-glycomic analyses of various biological samples.

Go To Anal Chem.

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For research to be discovered and acknowledged, it must be widely accessible and cited in a consistent and clear manner in the scientific literature. It’s a competitive environment for researchers today. Scientists and professors are constantly asked to create lots of supporting evidence that helps explain their work and its significance and impact to granting agencies, research institutions, peers and students. Indeed, funding agencies now often ask including references to public engagement (non-scientific audience). Their desire to measure and to improve the returns on their investments emphasizes accountability and dissemination

Global Medical Discovery evaluates and selects papers as key scientific articles on evidence of contribution, impact, peer-review and creativity. Global Medical Discovery can provide medical researchers with the tools to share the published research more widely and thus maximizing visibility. The featured authors will contribute a “significance Statement” which summarize the importance of their work in an understandable way to a large audience. An eye catching image can speak a thousand words is usually included. The image, usually never been published before, but it is possible to re-use a previously published image after taking permission from the publisher. Global Medical Discovery encourages the featured authors to say something about themselves and their research interests as well as an author photo. Global Medical Discovery will disseminate the medical research news on its portal, and a whole range of communications including social networks such as Twitter, Facebook, LinkedIn, Pinterest, Instagram and Google+.

Prostanoids regulate angiogenesis acting primarily on IP and EP4 receptors

Significance Statement

Angiogenesis is the new vasculature formation occurring in adult tissues. It is estimated that growth of any tissue to sizes above 2-3 mm³ requires formation of functional blood vessel network. Angiogenesis is regulated through a complex interplay of many factors, which either activate or inhibit the process. Participation of eicosanoids (large family of lipid mediators derived from arachidonic acid) in such regulation, although recognised as important, was never comparatively investigated in order to quantitatively assess contribution of individual eicosanoid class. In this work, we have determined (using in vitro endothelial cell migration and tube formation assays) that two eicosanoids, prostacyclin (or PGI₂) and prostaglandin E₂ (or PGE₂) are the main activators of angiogenesis acting via their receptors IP or EP4 respectively. The main eicosanoid regulator of angiogenesis is found to be prostacyclin which contributes to overall activation by 80%, whereas PGE₂ participates by approximately 20%. Better understanding of angiogenesis regulation could help with design of new drugs to prevent pathological angiogenesis observed in many diseases including cancer. 

About The Author

Dr Petrovic is a Senior Lecturer in Pharmacology in School of Pharmacy and Medical Sciences at University of South Australia. He has obtained his PhD at University of New York (USA) and post-doctoral training at Roswell Park Cancer institute (USA), University of Connecticut (USA) and University of Sydney (Australia).

Khuyen Gia Hoang is Honours student at University of South Australia.

Dr Sarah Allison is a postdoctoral researcher and Dr Michael Murray is a Professor of Pharmacology at University of Sydney.

Journal Reference

Microvasc Res. 2015;101:127-34.

Hoang KG¹, Allison S², Murray M², Petrovic N³.

Show Affiliations

1. School of Pharmacy and Medical Sciences, University of South Australia, Australia.
2. Pharmacogenomics and Drug Development Group, Discipline of Pharmacology, School of Medical Sciences, University of Sydney, Australia.
3. School of Pharmacy and Medical Sciences, University of South Australia, Australia. Electronic address: Nenad.Petrovic@unisa.edu.au.

Abstract

Angiogenesis is regulated by numerous activators and inhibitors, including prostanoids. Although many studies have identified their roles in inflammation, regulatory functions of prostanoids in angiogenesis are poorly understood. Here, we compared the activation of angiogenesis in vitro by two prostanoids with important vascular roles: prostaglandin E2 (PGE2) – thought to be the most important prostanoid activator of angiogenesis – and prostaglandin I2 (prostacyclin or PGI2), whose receptors are predominantly expressed in endothelial cells. Both of these prostanoids activate G-protein coupled receptors: EP1, EP2, EP3 and EP4 by PGE2 and IP by prostacyclin. Human umbilical vein endothelial cells (HUVECs) were used to characterize two pivotal pro-angiogenic processes in vitro: cell migration (using the matrigel droplet assay developed in our laboratory) and “tube formation” (a widely accepted method of assessing formation of blood vessel precursors). The suppression of cell migration and tube formation by the IP-specific antagonist CAY10441 was more extensive (~80%) than by the EP4-specific antagonist L-161,982 (~20%). AH6809, an antagonist of EP1, EP2 and EP3 receptors did not significantly suppress angiogenesis. Expression of the pro-angiogenic receptors KDR and Tie-2 in HUVECs was preferentially suppressed by antagonism of IP and EP4 receptors, respectively. EP4 and IP receptor agonists elicited biphasic actions on angiogenic processes in which there was activation at low concentration, and rapid desensitization at high concentrations – a characteristic common to many G-protein coupled receptors. Together these findings suggest that the prostacyclin-IP pathway plays a major role in the regulation of pro-angiogenic processes in HUVECs.

Copyright © 2015. Published by Elsevier Inc.

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Figure Legend Human umbilical vein endothelial cell structures used for migration and communication (filopodia, stained by fluorescent cholesterol stain filipin). Unpublished data from the study by Petrovic N. et al. Blood. 2007, 110: 142-150.

 

 

Adipose Derived Stromal Cell (ADSC) Injections for Pain Management of Osteoarthritis in the Human Knee Joint

Significance Statement

Osteoarthritis of the knee is the most common form of arthritis that cause pain, stiffness, and decreased function, and one of leading causes of disability. Stromal vascular fraction containing large amount of stem cells and other regenerative cells, can be easily obtained from loose connective tissue that is associated with adipose tissue. The authors concluded from this clinical study that Adipose-derived stem cell therapy for patients with knee osteoarthritis  might be effective in reducing pain, and improving function. Therefore, adipose-derived stem cell treatment appears to be a good option for osteoarthritis treatment.

Journal Reference

Aesthet Surg J. 2015 Aug 3.

Fodor PB, Paulseth SG.

Dr Fodor is an Associate Clinical Professor of Plastic Surgery, UCLA Medical Center, Los Angeles, California.

Dr Paulseth is an Adjunct Instructor of Clinical Physical Therapy, Division of Biokinesiology and Physical Therapy, University of Southern California, Los Angeles, California.

Abstract

BACKGROUND:

This safety and feasibility study used autologous adipose-derived stromal vascular cells (the stromal vascular fraction [SVF] of adipose tissue), to treat 8 osteoarthritic knees in 6 patients of grade I to III (K-L scale) with initial pain of 4 or greater on a 10-point Visual Analog Scale (VAS).

OBJECTIVES:

The primary objective of the study was evaluation of the safety of intra-articular injection of SVF. The secondary objective was to assess initial feasibility for reduction of pain in osteoarthritic knees.

METHODS:

Adipose-derived SVF cells were obtained through enzymatic disaggregation of lipoaspirate, resuspension in 3 mL of Lactated Ringer’s Solution, and injection directly into the intra-articular space of the knee, with a mean of 14.1 million viable, nucleated SVF cells per knee. Metrics included monitoring of adverse events and preoperative to postoperative changes in the Western Ontario and McMaster Universities Arthritis Index (WOMAC), the VAS pain scale, range of motion (ROM), timed up-and-go (TUG), and MRI.

RESULTS:

No infections, acute pain flares, or other adverse events were reported. At 3-months postoperative, there was a statistically significant improvement in WOMAC and VAS scores (P < .02 and P < .001, respectively), which was maintained at 1 year. Physical therapy measurements for ROM and TUG both improved from preoperative to 3-months postoperative. Standard MRI assessment from preoperative to 3-months postoperative showed no detectable structural differences. All patients attained full activity with decreased knee pain.

CONCLUSIONS:

Autologous SVF was shown to be safe and to present a new potential therapy for reduction of pain for osteoarthritis of the knee.

LEVEL OF EVIDENCE:

4 Therapeutic.

© 2015 The American Society for Aesthetic Plastic Surgery, Inc.

Go To Aesthet Surg J.

CD44 variant 6 is correlated with peritoneal dissemination and poor prognosis in patients with advanced epithelial ovarian cancer

Journal Reference

Cancer Sci. 2015;106(10):1421-8.

Tjhay F¹, Motohara T¹, Tayama S¹, Narantuya D¹, Fujimoto K¹, Guo J¹, Sakaguchi I¹, Honda R¹, Tashiro H², Katabuchi H¹.

Show Affiliations

1. Department of Obstetrics and Gynecology, Faculty of Life Sciences, Kumamoto University, Kumamoto City, Kumamoto, Japan.
2. Department of Maternal-Newborn Nursing, Kumamoto University, Kumamoto City, Kumamoto, Japan

Abstract

Cancer stem cells (CSCs) drive tumor initiation and metastasis in several types of human cancer. However, the contribution of ovarian CSCs to peritoneal metastasis remains unresolved. The cell adhesion molecule CD44 has been identified as a major marker for CSCs in solid tumors, including epithelial ovarian cancer. CD44 exists as a standard form (CD44s) and also as numerous variant isoforms (CD44v) generated by alternative mRNA splicing. Here we show that disseminated ovarian tumors in the pelvic peritoneum contain highly enriched CD44v6-positive cancer cells, which drive tumor metastasis and are responsible for tumor resistance to chemotherapy. Clinically, an increased number of CD44v6-positive cancer cells in primary tumors was associated with a shortened overall survival in stage III-IV ovarian cancer patients. Furthermore, a subpopulation of CD44v6-positive cancer cells manifested the ability to initiate tumor metastasis in the pelvic peritoneum in an in vivo mouse model, suggesting that CD44v6-positive cells show the potential to serve as metastasis-initiating cells. Thus, the peritoneal disseminated metastasis of epithelial ovarian cancer is initiated by the CD44v6-positive subpopulation, and CD44v6 expression is a biomarker for the clinical outcome of advanced ovarian cancer patients. Given that a distinct subpopulation of CD44v6-positive cancer cells plays a critical role in peritoneal metastasis, definitive treatment should target this subpopulation of CD44v6-positive cells in epithelial ovarian cancer.

© 2015 The Authors. Cancer Science published by Wiley Publishing Asia Pty Ltd on behalf of Japanese Cancer Association.

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Recent advances in the study of live attenuated cell-cultured smallpox vaccine LC16m8

Significance Statement

Smallpox was one of the most feared infectious diseases in the world. It is caused by the variola virus, an orthopoxvirus, which results in high fever and progressive rashes all over the body, killing up to 30% of the infected patients.
In the 1960s, World Health Organization (WHO) launched a global eradication campaign using first-generation smallpox vaccines, consisting of vaccinia virus, and declared the eradication of smallpox in 1980. Consequently, routine vaccination programs were discontinued.However, owing to terrorist events in the USA in the beginning of this century, the need for the smallpox vaccine had to be reconsidered. Several countries have since stockpiled the smallpox vaccine as a measure against bioterrorism.
Since 2002, Japan has re-produced and stockpiled LC16m8, a live attenuated third-generation vaccine, which was developed as a safer alternative in the 1970s because of the adverse effects of the first-generation smallpox vaccines.

Repeated evaluations of LC16m8 have demonstrated an excellent safety profile, and an efficacy comparable to that of first-generation vaccines. Severe adverse events attributable to the vaccine have not been reported so far, and the vaccine has been safely administered even to immunodeficient animals. Additionally, LC16m8 has beneficial properties for practical use: induction of visible major cutaneous reactions, signs of successful vaccination, and easy administration using bifurcated needles.

Detailed information on the safety and efficacy of LC16m8 is critical for public health decision-makers to plan strategic vaccination programs in case of a possible emergent outbreak of smallpox. Even after eradication, possibilities of a re-occurrence of the disease exist, under intended or unintended circumstances. Therefore, a vaccine stockpile and updated research are necessary public health measures.

An established vaccine may also be useful against related diseases. The routine vaccination of smallpox vaccines were effective against monkeypox, another disease caused by orthopoxvirus in Africa. The discontinuation of the smallpox vaccination program has decreased the immunity of populations against both smallpox and monkeypox, leading to concerns regarding possible outbreaks of the disease; thus, preparedness for these possibilities is desirable.

Although LC16m8 conferred protective effects against monkeypox in macaques, its efficacy against monkeypox in humans has not been evaluated. Additionally, LC16m8 has not been evaluated in immunocompromised patients. Further studies are expected for the evaluation of LC16m8 in these applications.

 

Journal Reference

Vaccine. 2015. pii: S0264-410X(15)01179-2.

Eto A¹, Saito T¹, Yokote H², Kurane I³, Kanatani Y⁴.

Show Affiliations

1. Department of Health Crisis Management, National Institute of Public Health, 2-3-6 Minami, Wako-shi, 351-0197, Saitama, Japan.
2. Chemo-Sero-Therapeutic Research Institute (Kaketsuken), 1-6-1 Okubo, Kita-ku, Kumamoto-shi, 860-8568, Kumamoto, Japan.
3. National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, 162-8640, Tokyo, Japan.
4. Department of Health Crisis Management, National Institute of Public Health, 2-3-6 Minami, Wako-shi, 351-0197, Saitama, Japan. Electronic address: ykanatani@niph.go.jp.

Abstract

LC16m8 is a live, attenuated, cell-cultured smallpox vaccine that was developed and licensed in Japan in the 1970s, but was not used in the campaign to eradicate smallpox. In the early 2000s, the potential threat of bioterrorism led to reconsideration of the need for a smallpox vaccine. Subsequently, LC16m8 production was restarted in Japan in 2002, requiring re-evaluation of its safety and efficacy. Approximately 50,000 children in the 1970s and about 3500 healthy adults in the 2000s were vaccinated with LC16m8 in Japan, and 153 adults have been vaccinated with LC16m8 or Dryvax in phase I/II clinical trials in the USA. These studies confirmed the safety and efficacy of LC16m8, while several studies in animal models have shown that LC16m8 protects the host against viral challenge. The World Health Organization Strategic Advisory Group of Experts on Immunization recommended LC16m8, together with ACAM2000, as a stockpile vaccine in 2013. In addition, LC16m8 is expected to be a viable alternative to first-generation smallpox vaccines to prevent human monkeypox.

Copyright © 2015 Elsevier Ltd. All rights reserved.

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Neutrophil-to-lymphocyte ratio and overall survival in all sites of head and neck squamous cell carcinoma

Significance Statement

Biomarkers such as Neutrophil and lymphocyte counts, neutrophil-to-lymphocyte ratio (NLR) have been proposed as potential prognostic factors for cancer. There is accumulating evidence for the association of NLR with survival of patients with many kinds of cancers. In order to obtain an objective and consistent conclusion, The authors conducted this comprehensive analysis of the association between NLR and survival of oral, pharyngeal, and laryngeal squamous cell carcinomas.

Journal Reference

Head Neck. 2015 . doi: 10.1002/hed.24159.

Rachidi S^1,2, Wallace K^2,3, Wrangle JM^1,4, Day TA^2,5, Alberg AJ^2,3, Li Z^1,2,4.

Show Affiliations

1. Department of Microbiology and Immunology, Medical University of South Carolina, Charleston, South Carolina.
2. Hollings Cancer Center, Medical University of South Carolina, Charleston, South Carolina.
3. Department of Public Health Sciences, Medical University of South Carolina, Charleston, South Carolina.
4. Division of Hematology and Oncology-Department of Internal Medicine, Medical University of South Carolina, Charleston, South Carolina.
5. Head and Neck Tumor Center, Department of Otolaryngology-Head and Neck Surgery, Medical University of South Carolina, Charleston, South Carolina.

Abstract

BACKGROUND:

Current prognostic criteria are insufficient in predicting outcomes in head and neck cancer, necessitating new, readily available biomarkers.

METHODS:

Pretreatment neutrophil and lymphocyte counts and their ratio (NLR) were retrospectively investigated for correlation with overall survival while controlling for demographic and clinical confounders.

RESULTS:

Patients in the highest tertile of neutrophil counts and those in the lowest tertile of lymphocytes experienced shorter survival than the rest of the population. Patients in the highest tertile of the neutrophil-to-lymphocyte ratio were at a higher risk compared with those in the lowest tertile after multivariate analysis (hazard ratio [HR] = 2.39; p = .0001). Additionally, NLR was lower in patients with human papillomavirus (HPV)-positive tumors compared to HPV-negative tumors and predicted survival in both tumor types.

CONCLUSION:

Neutrophil and lymphocyte counts are strong biomarkers with opposing prognostic significance and the NLR is a robust predictor of overall survival in oral, pharyngeal, and laryngeal squamous cell carcinomas. © 2015 Wiley Periodicals, Inc. Head Neck, 2015.

© 2015 Wiley Periodicals, Inc.

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Visible assay for glycosylase based on intrinsic catalytic ability of graphene/gold nanoparticles hybrids

Journal Reference

Biosens Bioelectron. 2015; 68:7-13.

Yuan F¹, Zhao H², Liu M¹, Quan X¹.

Show Affiliations

1. Key Laboratory of Industrial Ecology and Environment Engineering (Ministry of Education, China), School of Environmental Science and Technology, Dalian University of Technology, Dalian 116024, China.
2. Key Laboratory of Industrial Ecology and Environment Engineering (Ministry of Education, China), School of Environmental Science and Technology, Dalian University of Technology, Dalian 116024, China. Electronic address: zhaohuim@dlut.edu.cn.

Abstract

A sensitive, rapid and label-free assay for colorimetric detection of human 8-hydroxyguanine glycosylase (hOGG1) was proposed based on the tunable catalytic ability of graphene/gold nanoparticles (graphene/Au-NPs) hybrids and the terminal protection of hOGG1. In presence of H2O2, the hybrids were capable of catalyzing the oxidation of color developing reagent, causing a concomitant change in color. Due to the excellent controllability, the capacity could be inhibited by adsorption of ssDNA onto the hybrids sheets and recovered when the adsorbed ssDNA was digested by exonuclease. The terminal protection of human 8-hydroxyguanine glycosylase could irreversibly interrupt the digestion of the captured ssDNA (containing the oxidative damage site) by the exonuclease, thus preventing the catalytic  ability  of graphene/Au-NPs from being recovered. The original color change which related to the concentration of the protected ssDNA facilitated quantitative detection of human 8-hydroxyguanine glycosylase activity. Compared with conventional methods for human 8-hydroxyguanine glycosylase detection, the presented assay without any labeling process greatly simplified the operation steps and reduced the analysis time. This approach performed a linear response for human 8-hydroxyguanine glycosylase activity from 0.02 to 0.11 U/μL with a detection limit of 0.0016 U/μL, and realized the quantification of human 8-hydroxyguanine glycosylase activity in real cell lines.

Copyright © 2015 Elsevier B.V. All rights reserved.

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Characterization of a psoriatic skin model produced with involved or uninvolved cells

Journal Reference

J Tissue Eng Regen Med. 2015;9(7):789-98.

Jean J^1,2, Leroy M^1,3, Duque-Fernandez A^1,2, Bernard G¹, Soucy J⁴, Pouliot R^1,2.

Show Affiliations

1. Laboratoire d’Organogénèse Expérimentale, Centre de Recherche FRSQ du CHU de Québec, Canada.
2. Faculté de Pharmacie, Université Laval, Québec, Canada.
3. Laboratoire d’Ingénierie de Surface (LIS), Département de Génie des Mines, de la Métallurgie et des Matériaux, Centre de Recherche sur les Matériaux Avancés, Université Laval, Québec, Canada.
4. Département de Dermatologie, Hôpital de l’Enfant-Jésus, Québec, Canada.

Abstract

Current knowledge suggests that uninvolved psoriatic skin could demonstrate characteristics associated with both normal and involved psoriatic skins. However, the triggering factor allowing the conversion of uninvolved skin into a psoriatic plaque is not fully understood. To counter this lack of information, we decided to develop pathological skin substitutes  produced with  uninvolved psoriatic cells, in order to better characterize the uninvolved psoriatic skin. Substitutes were produced using the self-assembly approach. Macroscopic, immunohistochemical, permeability and physicochemical results showed that involved substitutes had a thicker epidermis, higher cell proliferation, abnormal cell differentiation and a more permeable and disorganized stratum corneum compared with normal substitutes. Various results were observed using uninvolved cells, leading to two proposed profiles: profile 1 was suggested for  uninvolved skin substitutes mimicking the results obtained with normal skin substitutes; and profile 2 was dedicated to those mimicking  involved skin substitutes in all aspects that were analysed. In summary, uninvolved substitutes of profile 1 had a thin, well-organized epidermis with normal cell proliferation and differentiation, such as observed with normal substitutes, while uninvolved substitutes of profile 2 showed an inverse trend, i.e. a thicker epidermis, higher cell proliferation, abnormal cell differentiation and a more disorganized and more permeable stratum corneum, such as seen with involved substitutes. The results suggest that uninvolved substitutes could demonstrate characteristics associated with both normal or involved psoriatic skins.

Copyright © 2012 John Wiley & Sons, Ltd.

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