Optogenetics-Mediated Ca²⁺ Signaling in Regulation of Ca²⁺- Dependent Transcription Factor Activation

Wen-Tai Chiu^*, Ya-Han Chang and Yi-Shyun Lai

National Cheng Kung University, Taiwan

The ability of a simple ion such as Ca²⁺ to play an highly versatile intracellular signal results from the facility that cells have to shape Ca²⁺ signals in the dimensions of space, time and amplitude. Thus, spatial and temporal changes in intracellular Ca²⁺ concentration is important to determinate the cell fate. Optogenetics is the combination of genetics and optics to control well-defined events within specific cells of living tissue in real time. Optogenetic stimulation approach that uses channelrhodopsin-2 (ChR2) related proteins has been developed for providing more precise and targeted stimulation effect on cells in vivo and in vitro. We aimed to pose the formerly unaddressed fundamental question of how Ca²⁺ signals regulate transcription factor activation by manipulating intracellular Ca²⁺ through using optogenetic strategies. Three Ca²⁺-dependent transcription factors, NFAT, NFkB and CREB, were examined in this study. We found that (1) elevation of intracellular Ca²⁺ levels that correspond with the power, frequency and duty cycle of light illumination in a dose-dependent manner; (2) activation of Ca²⁺-dependent transcription factors is depends on Ca²⁺ oscillations with different frequency and amplitude in ChR2 overexpressed cells. Moreover, activation of NFκB required lowfrequency and low-amplitude Ca²⁺ oscillations, whereas high-frequency and high-amplitude Ca²⁺ oscillations tended to activate NFAT. On the other hand, CREB activation occurred regardless of the frequency and amplitude of Ca²⁺ oscillations. In summary, the delicate regulation and precise control of transcription factor activation can be achieved under the optogenetic platform. It will enable us to be confident in applying this advanced technique to manipulate the expression patterns of genes by desire. Such a process would be a potential platform in study of cell development and cancer.

Biography:
Dr. Wen-Tai Chiu received the Bachelorʼs, Masterʼs and Ph.D. degrees from National Cheng Kung University, Taiwan in 1997, 1999 and 2007, respectively. He has been a postdoctoral fellow at the University of Texas MD Anderson Cancer Center from 2008 to 2010. Prof. Chiu is currently an associate professor in the Department of Biomedical Engineering at National Cheng Kung University. His research interests lie in the area of Ca2+ signaling and molecular imaging of cancers. Much of his work has been on improving the understanding, design and performance of Ca2+ in focal adhesion dynamics, cell migration, metastasis and chemoresistance.

Long-Term Results of Anorectal Physiologic Changes and Recurrence between Transanal Repair and Transanal Repair with Posterior Colporrhaphy in Patients with Symptomatic Rectocele

Joo Hyung Kim^1*, Dae Hyun Kim² and Yong Pyo Lee³

¹Ajou University School of Medicine, South Korea
²University of Alabama, USA
³

Hanvit Hospital, South Korea

Background: Rectoceles are often associated with anorectal symptoms. Various surgical techniques have been described to repair rectoceles, but the surgical results vary. The aim of this study was to compare transanal repair (TAR) and transanal repair with posterior colporrhaphy (TAR+PC).

Methods: While 44 patients went through TAR, the other 49 patients went through TAR+PC. Patients were followed up with anorectal physiological studies three month post-surgeries. 22 patients who went through TAR and 25 patients who went through TAR+PC agreed to participate in a three year post-treatment check-up.

Results: Out of 22 patients that went through TAR, 3 of them (13.6%) scored higher than 15 on their constipation scoring system (CSS) 3-month post-treatment while 1 out of 25 patients that went through TAR+PC scored higher than 15 on their CSS 3-month post-treatment, which is considered as recurrence (p=0.237). With 7 patients from TAR treatment group (31.8%) and 2 patients from TAR+PC treatment group (8.0%) showing recurrence of rectocele at 3-year post-treatment observation, this research has found that TAR+PC resulted in much lower recurrence rate, when compared with TAR treatment method. In rectal sensation, sensory threshold (p=0.001) and early defecation urge (p=0.003), this research has shown that TAR+PC is more effective treatment method than TAR.

Conclusions: TAR+PC can help alleviate some symptoms through revitalizing the rectal sense and improving the rectovaginal septum. In other words, it is believed that addition of a simple treatment method can ultimately lead to lowering of a recurrence rate of rectocele.

Biography:
Dr. Joo Hyung Kim is a colorectal surgeon, graduated from Soonchunhyang University School of Medicine, South Korea in 1993. In 2006, he earned a Ph.D. from the Graduate School, Ajou University, South Korea. He is currently a professor at department of surgery, Ajou University School of Medicine. He prized Scientific Award of the Korean Society of Coloproctology in 2004 and 2008. He is currently active as a Member of American Society of Colorectal Surgeon (USA), Member of International Society of University Colon & Rectal Surgeons (USA) and Board member of directors of the Korean Society of Coloproctology (Republic of Korea).

Experience of Synthetic Analogues of Somatostatine from Esophageal Variceal Bleeding in Patients with Hepatic Cyrrosis with Syndrome of Portal Hypertension in the Multidisciplinary Surgical Center of Republic Sakha (Yakutia)

Mikhail Mikhailovich Vinokurov^*, V V Saveliev, V P Egorova, T V Yalynskaya and L M Vinokurova

North-Eastern Federal University, Russia

The aim of the study was to evaluate the effectiveness of vasoactive therapy with synthetic somatostatin analogues from acute esophageal variceal bleeding in patients with hepatic cirrhosis with portal hypertension syndrome.

Material and Methods: The study is based on a retrospective analysis of the results of a comprehensive treatment of 57 patients with cirrhosis and portal hypertension syndrome who were on treatment at the emergency surgical department of the Republican Hospital №2 − the Center for Emergency Medical Care of the Republic of Sakha (Yakutia) in the period from 2015 to 2017. All patients were divided into two groups. The first group consisted of 23 (40.4%) patients, who only used the Sengstaken-Blakemore probe to stop from acute esophageal variceal bleeding. The second group of the study comprised 34 (59.6%) patients who, together with the Sengstaken-Blakemore probe installation, used synthetic somatostatin analogues in complex intensive therapy.

Results: The presented clinical experience of the use of synthetic somatostatin analogues in complex therapy from acute esophageal variceal bleeding to recommend their wide use in the practice of urgent surgical clinics.

Conclusion: The introduction of complex intensive therapy from acute esophageal variceal bleeding in patients with hepatic cirrhosis with the syndrome of portal hypertension of synthetic somatostatin analogues allowed to reduce the overall lethality to 10.7% and to reduce the risk of rebleeding by 17%.

Keywords: Hepatic Cirrhosis, Portal Hypertension, Esophageal Variceal Bleeding, Vasoactive Therapy.

Biography:
Dr. Mikhail Mikhailovich Vinokurov is a Professor, Head at the Department of Faculty Surgery, Urology, Oncology and Otorhinolaryngology, Medical Institute of the Federal State University, North- Eastern Federal University named after M.K Ammosov, Russia.

Regeneration Strategy in Medicine using Whartonʼs Jelly Mesenchymal Stem Cells

Aleksandra Musiał-Wysocka^*, Maciej Sułkowski, Marta Kot and Marcin Majka

Jagiellonian University Medical College, Poland

Whartonʼs Jelly is a special type of connective, gelatinous tissue located in the umbilical cord. Mesenchymal Stem Cells isolated from Whartonʼs Jelly (WJ-MSCs) are advantageous for cell-based therapy because of their self-renewal capacity in vitro, high plasticity and low immunogenicity. In addition, WJ-MSCs could be very useful for clinical application because they inhibit the immunological response and does not need a major histocompatibility match for allogeneic transplantation. Due to their properties i.e., the secretion of many proangiogenic factors, WJ-MSCs present an opportunity for effective regeneration of ischemic muscles by promoting endogenous angiogenesis.

The WJ-MSCs show the minimal criteria outlined for MSC by the International Society for Cellular Therapy. More than 90% of the population expresses specific mesenchymal markers such as CD90, CD 105, CD 73. Furthermore, WJ-MSCs are negative for hematopoietic antigens- CD45, CD3. The expression levels of specific surfaces markers have been analyzed using flow cytometry technique. The expression levels of specific proangiogenic factors (VEGF A, ANGPT, MMP-1, HIF 1α, FGF 2, HGF) in WJ-MSCs was analyzed using Real-Time PCR.

In our study, we created a murine hindlimb ischemia model to test a new method of treating this disease and evaluate the regenerative capacity of Mesenchymal Stem Cells isolated from Whartonʼs Jelly. The vascular response in hindlimb ischemia model was evaluated by Laser Speckle-based perfusion measurements as well as histological quantification of arteriogenesis and angiogenesis. The results of our study have confirmed as well as the high differentiation potential of WJ-MSCs and their proangiogenic character that promote angiogenesis.

This work was supported by research grant (STRATEGMED2/265761/10/NCBR/2015) from the National Center for Research and Development.

Biography:
Dr. Aleksandra Musiał-Wysocka is a PhD student at Department of Transplantation Institute of Pediatrics Jagiellonian University Medical College in Cracow. The subject of her research is the characteristic of mesenchymal stem cells isolated from Whartonʼs Jelly and the possibility of using them in regenerative medicine. The main goal of her study is to develop a new therapy for cardiovascular diseases using stem cells.