Madridge Journal of Cancer Study & Research

ISSN: 2640-5180

5th International Conference on Oncology & Virology

July 25-26, 2019, Holiday Inn Rome Aurelia, Rome, Italy
Poster Session Abstracts
DOI: 10.18689/2640-5180.a4.008

Cowpea Aphid-Borne Mosaic Virus does not seen to be transmitted by Hand Pollination of Passion Flower

David Marques de Almeida Spadotti* and Jorge Alberto Marques Rezende

ESALQ/University of São Paulo, Brazil

To maximize pollination, which has direct effect on the increase of fruit yield, Brazilian passionfruit producers adopt artificial hand pollination in the orchards. This is done with the tips of the fingers, transferring pollen among flowers. Passionflower orchards are severely affected by the potyvirus Cowpea aphid-borne mosaic virus (CABMV), which is naturally transmitted by aphids. As this virus is easily and efficiently transmitted mechanically, the question arises whether there is a possibility of being transmitted through small wounds that can be produced in the flowers during hand pollination. Therefore, the aim of this work was to evaluate the transmission of CABMV by means of hand pollinating flowers of cultivar FB-200 and mechanically transmit the virus to passion fruit test-plants with extracts of anther and pollen collected from flowers of infected plants. Infection was assessed by means of symptoms and virus detection by PTA-ELISA and RT-PCR. Flowers of eight healthy plants, grown in a greenhouse, were pollinated for 60 days with pollen from CABMV-infected flowers. Two hundred and one flowers were pollinated, 68 fruits were obtained, and no plants were infected with the potyvirus. Extracts from CABMV-infected anthers and pollen, separately inoculated mechanically on 35 passionfruit test-plants resulted in infection of three and zero plants, respectively. Studies are under way to detect CABMV in anther and pollen samples by RT-PCR and RT-qPCR. In conclusion, CABMV hand pollination does not appear to be efficient for CABMV transmission in passionflower orchards.

Biography:
David Marques de Almeida Spadotti was Bachelorʼs degree in Agronomy, UNESP (2010). Masterʼs degree in Plant Pathology, USP (2012), working on biological, molecular and serological studies of Zucchini yellow mosaic virus. PhD in Plant Protection, UNESP (2016) focusing on tospo virus plant diseases. Currently as postdoctoral fellow at USP, working in the management of viruses of passion flower.

Study of HLA-C Binding Stability in HIV-1 Infection and in Cognitive Disorders

Chiara Stefani*, Simona Mutascio, Stefania Fochi, Maria Grazia Romanelli and Donato Zipeto

University of Verona, Italy

MHC-I is an heterotrimeric complex composed by HLA-C/β2-microglobulin/peptide. The different HLA-C variants can be grouped into stable and unstable clusters based on their binding stability to β2-microglobulin. The presence of unstable HLA-C molecules increases β2-microglobulin release. It is known that patients affected by AIDS Dementia Complex (ADC), a very severe neurological condition of HIV-1 infection, present high level of β2-microglobulin in the cerebrospinal fluid. We observed a higher frequency of unstable HLA-C alleles in ADC patients. We demonstrated that, upon HIV-1 infection, HLA-C molecules associate with HIV-1 virions, increasing viral infectivity. In addition, HIV-1 virions produced in the presence of unstable HLA-C variants are more infectious.We aimed to evaluate how each HLA-C variant affects the ADC onset in HIV-1 infected patients. To assess the contribution of each HLA-C variant in the modulation of HIV-1 infection, CRISPR/Cas9 was used to generate 293T HLA-C-/-packaging cells. The different HLA-C alleles were transfected in 293T HLA-C-/- cells, to develop different cell lines expressing a specific HLA-C allotype. The different cell lines will be used to produce HIV-1 pseudotyped viruses to be tested in infectivity assays conducted on TZm-bl cells. Furthermore, to assign stability score to each HLA-C variant, based on their dissociation rate from β2-microglobulin, each HLA-C expressing cell line will be treated with an acid wash, to study the kinetic of β2-microglobulin dissociation from HLA-C. These analyses will be fundamental to clarify the relationship between HLA-C binding stability, HIV-1 infection progression and the development of HIV-1 related neurocognitive diseases.

Porcine Reproductive and Respiratory Syndrome Viruses (PRRSVs) Attenuated by Synthetic-Attenuated Virus Engineering (SAVE) in NSP1 Induced Protective Immunity in Pigs

Sang-Ho Cha1* and Changhoon Park2

1APQA, South Korea
2BioPOA, South Korea

A computer-based codon-pairs deoptimization technology, synthetic attenuated virus engineering (SAVE), is utilized to lower virulence of various viruses. The rapid viral attenuation concept by the codon pair deoptimization is to decrease codon pair bias (CPB), which demonstrated the potential usefulness as a technology to develop live attenuated vaccine. For this, we produced field PRRSVs attenuated by codon-pair deoptimization. The affection of SAVE on virulence of the attenuated strains was confirmed in pigs. Type 2 porcine reproductive and respiratory syndrome viruses (PRRSVs) isolated from PRRS-affected swine farms in South Korea were attenuated by de-optimization of codon pair bias in NSP1. In animal infection using 3 weeks-old pigs, the attenuated viruses showed significantly lowered replication ability than the parental viruses without distinct clinical sign and pathological lesions, which were observed in pig infected with the parental viruses. Regarding induction of PRRSV specific immunity, the level of the neutralizing antibodies as well as secretion of IFN-γ-SCs in PBMCs was not different between the attenuated viruses and the parent viruses. Conclusively, the attenuated viruses generated by SAVE in this study demonstrated the potential usefulness as vaccine strains to provide immune protection against diverse PRRSVs

Biography:
Dr. Cha has his expertise in Animal virology and passion in developing vaccine and diagnostics against animal viral diseases to improve economy of livestock industry as well as welfare of pet animals. He has many experiences in research about developing measures to minimize outbreak of animal diseases. Also, he has a big interest in combining high advanced technology on the research area.

Structure and Oligomerization State of the C-Terminal Region of the Middle East Respiratory Syndrome Coronavirus Nucleoprotein

Thi Hong Van Nguyen1,2*, Julie Lichière1,2, Bruno Canard1,2, Nicolas Papageorgiou1,2, Sarah Attoumani1,2, François Ferron1,2 and Bruno Coutard1,2

1Aix-Marseille Université, France
2CNRS, France

Middle East respiratory syndrome (MERS) is a positive single-stranded RNA virus belonging to lineage C of the genus Beta-Coronavirus within the Corona viridae family and the order Nidovirales (Chan et al., 2015). In 2015-2016, there was a largest outbreak seen in Middle East, and the Republic of Korea with the mortality rate up to 35%. MERS-CoV is a very dangerous respiratory disease, but currently, there is no vaccine or specific treatment to be available. Treatment is supportive and based on the patientʼs clinical condition (WHO, Jan 2018). Therefore, it is necessary to have more researches on it to find an antiviral drug for treatment. The nucleoprotein (N) structure of virus in the genus alpha-CoV, gamma-CoV and beta-CoV lineage B have identified, but the N structure of MERS-CoV has not identified yet. The Coronavirus nucleoprotein (CoV N) includes two folded domains called N-terminal domain (NTD) and C-terminal domain (CTD), separated by a linker region. CoV N has many functions, such as interacting with structural proteins; packaging the viral genome with structural proteins to form ribonucleo protein complexes; stopping host translation and making immune system interference. The main function of CTD nucleoprotein (N CTD) is a strong candidate for design of diagnostic tools (Ruth McBride et al., 2014; Chan et al., 2015).The structure of NTD was identified, but there is no structure for CTD and full length N MERS-CoV in order to support this functional hypothesis. Therefore, the aim of project is to characterize the N CTD structure of MERS-CoV using X-ray crystallography.

Biography:
Nguyen Thi Hong Van is a Vietnamese student. In 2014, she performed the 1st Master Internship in Hanoi University of Pharmacy, Vietnam. In 2015, she carried out the 2nd Master Thesis in Institute of Sophia Agrobiotech, Nice Sophia-Antipolis University, France. Then, she completed her Masters at the University of Sciences and Technologies of Hanoi, Vietnam and at the Co-University of Montpellier II, France. From 2016 until now, she has been a Doctoral student who has worked in Architecture et Fonction des Macromolécules Biologiques (AFMB), Marseille, France.

Investigation of Hemorrhagic Fever Viruses inside Wild Populations of Ticks: One of the Pioneer Studies in Saudi Arabia

Areej Abdulkareem Al Khalaf1*, Rania Ali Mohamed1, Nahla Mohamed1, Fadilah Sfouq Aleanizy2, Fulwah Yahya Alqahtani1 and Lamya Ahmed AlKeridis1

1Princess Nourahbint Abdulrahman University, Kingdom of Saudi Arabia
2King Saud University, Kingdom of Saudi Arabia

Objective: To screen hemorrhagic fever viruses inside wild populations of ticks collected from Riyadh, Saudi Arabia between January and March 2016.

Methods: Ticks were identified depending on their morphological features using classical keys then grouped into pools. Ticks in each pool were processed separately using the sterile pestles and mortars. Viral RNA was extracted using Qiagen RNeasy Mini Kit and Qiagen RNAeasy Columns (Qiagen, Hilden, Germany) according to the instructions of manufacturers. A total number of 1, 282 hard ticks were collected, and 582 of them were precisely identified then screened for the presence of arboviruses using quantitative real-time PCR. The four species were screened for six viruses: Rift Valley fever virus (RVFV), Chikungunya virus (CHIKV), Crimean-Congo hemorrhagic fever virus (CCHFV), Alkhurma virus (INKV), Sindbis virus (SINV), and Pan Hanta virus (HANTA). CT value for the negative control (RNA free water) was zero. Negative and positive controls were tested for each test to confirm the specificity of the selected primer pairs. SYBR Green One step RT-PCR Master Mix (KAPA Biosystems, Boston, MA) was tested along with primers.

Results: Ticks identification resulted into four species: Hyalommaschulzei, Hyalommaonatoli, Boophiluskdhlsi and Hyalommdromedarii. All the ticksʼ species (except Boophiluskdhlsi) were positive for the following viruses: SINV, RVFV, CHIKV, and CCHFV. While HANTA viruses have been detected in a single species (Hyalommdromedarii).

Conclusions: According to our knowledge this research may be one of the pioneer studies in Kingdom of Saudi Arabia. Incrimination of the above-mention edticks species as well as their vectorial capacity is highly recommended for investigation in the upcoming researches.

Biography:
Dr. Areej Abdulkareem Alkhalaf is associate professor of entomology at Princess Nourahbint Abdulrahman University (Biology department) Riyadh, Saudi Arabia. She is a member in some scientific societies such as Entomological Society of America. She has more than 20 published scientific researches. Her research focuses on insectʼs biology, histology, behavior, molecular, classification, ecology microbial control and pest management. She participated and attended many of the internal and external conferences in the area of specialization, bio nanotechnology, e-learning, quality, planning, leadership and management. She has held several managerial positions during her career and research. She was the Dean of Scientific Research at PNU. Head of Biology Dept. Vice Head of Zoology Dept. Quality Control Officer. Chairperson of the Environment Committee, Scientific Research Co-coordinator at College of Science, Chief of the Biology Department laboratories.

Development of IFN-Deficient System to Manufacture IFN-Sensitive Influenza Vaccine Virus

Lei Sun*, Can Chen and Wenjun Liu

CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, China

Interferon (IFN)-sensitive influenza viruses are likely to be the alternatives to inactivated and attenuated virus vaccines. Some IFN-sensitive influenza vaccine candidates with modified non-structural protein 1 (NS1) are highly attenuated in IFN-competent hosts but induce robust antiviral immune responses. However, little research has been done on the manufacturability of these IFN-sensitive vaccine viruses.

In this study, RIG-I-knockout 293T cells were used to package the IFN-sensitive influenza A virus expressing the mutant NS1 R38A/K41A. We found that the packaging efficiency of the NS1 R38A/K41A virus in RIG-I-knockout 293T cells was much higher than that in 293T cells. Moreover, the NS1 R38A/K41A virus could no longer replicate in A549, MDCK, and Vero cells after 3-6 passages, indicating that the replication of NS1 R38A/K41A virus is limited in conventional cells. Therefore, we further established a stable Vero cell line expressing the wild-type (WT) NS1based on the Tet-On 3G system. The NS1 R38A/K41A virus was able to steadily propagate in the NS1-overexpressing Vero cell line with high titers for at least 20 passages. Additionally, the replication of NS1 R38A/K41A virus was limited in mice but triggered strong antiviral immune responses.

In conclusion, we presented a promising IFN-deficient system, involving a RIG-I-knockout 293T cell line to package the IFN-sensitive vaccine virus and a stable NS1-overexpressing Vero cell line to propagate the IFN-sensitive vaccine virus. The IFN-deficient system is applicable for the manufacture of IFN-sensitive and replication-limited vaccine virus.

Biography:
Lei Sun obtained her doctoral degree from the Yangzhou University. Dr. Sun is an associate professor working at CAS Key Laboratory of Pathogenic Microbiology of Immunology, Institute of Microbiology, Chinese Academy of Science. Her research focuses on the replication and pathogenesis of influenza virus, the post-translational modification of viral protein, and the antiviral immunity. The research works are designed to increase fundamental knowledge as well as to facilitate the development of new approaches to control of viral infection.

Identification and Characterization of Nuclear Export Signals in Influenza B Virus Nuclear Export Protein

Wenhui Fan1*, Pengtao Jiao1,2, Lei Sun1 and Wenjun Liu1

1Institute of Microbiology, Chinese Academy of Sciences, China
2College of Animal Sciences and Veterinary Medicine, Guangxi University, China

The nuclear export signal (NES) of influenza A virus nuclear export protein (NEP) has been proposed to mediate the nuclear export of NEP and viral ribo nucleo protein (vRNP). However, much less is known about the nuclear export mechanism of influenza B virus NEP. In the present study, we identified three leucine-rich NESs (NES1, NES2, and NES3) in influenza B virus NEP. The intracellular distributions of green fluorescent protein (EGFP)-tagged wild type and NESs-mutant NEPs were determined. The results showed that the nuclear export of the NES1/NES2/NES3-mutant NEP was significantly decreased compared with that of the wild type NEP, suggesting that all three NESs are critical for NEP nuclear export. Furthermore, these three NESs regulated NEP nuclear export through CRM1-dependent manner and were necessary for influenza B virus replication. This study expands our understanding of the life cycle of influenza B virus by defining the nuclear export mechanism of influenza B virus NEP and could provide a scientific basis for the development of anti-influenza drugs.

Biography:
Dr. Wenhui Fan is an Assistant Professor at CAS Key Laboratory of Pathogenic Microbiology of Immunology, Institute of Microbiology, Chinese Academy of Science. She obtained her master degree from Ocean University of China in 2005 and got her doctoral degree from Guang Xi University in 2015. Her research focuses on the pathogenic mechanism of influenza A and B viruses, virus-host interaction, and development of antiviral drugs and biological products.

Anti-Rota Virus PEGylated Proteins from Bifidobacterium adolescentis Secretome do not Affect Viability, Tight Junction Integrity and Cytoskeleton Organization of Human Intestinal Cells - C2BBe1

Juan Carlos Ulloa Rubiano1*, Martin Sieger1, Alfonso Barreto2 and María Hernandez1

1Laboratorio de Virología, Facultad de Ciencias, Grupo de Investigación en Enfermedades Infecciosas, Pontifical Javeriana University, Colombia
2Grupo de Inmunobiología y Biología Celular, Facultad de Ciencias, Pontificia Universidad Javeriana, Colombia

Background: Acute diarrheal disease (ADD) remains a public health problem worldwide, causing 525.000 deaths of children under 5years of age and 1.7 billion cases childhood diarrheal disease every year. Rotavirus is the most common etiological agents and causes approximately 5% of the deaths of children under 5years of age. The treatment used to control ADD produced by rotavirus is not specific and although two vaccines are available, these do not prevent the viral infection. Other important problems include lack of access in low income countries and genetic re assortment between vaccine strains and wild-type strains. As an alternative, probiotics bacteria have been used to prevent the rotavirus infection and reduce the duration and severity of diarrhea. The probiotic Bifidobacterium adolescentis produces some proteins in the extra cellular secretome which exert in vitro anti-rotavirus activity when they are PEGylated. The potential use of these PEGylated proteins for oral administration in humans should be established using initially in vitro models of human intestinal cells.

Main Goal: To evaluate the effects exerted by anti-rotavirus PEGylated proteins produced in the B. adolescentis extra cellular secretome on the cellular viability, tight junction integrity and cytoskeleton organization of human intestinal cells - C2BBe1.

Methods: The effect exerted on the viability of C2BBe1 cells by 14 concentrations of PEGylated proteins previously obtained from the extra cellular secretome of B. adolescentis was measured 24 hours post exposure by the MTT method at 540 nm. Likewise, the tight junctions (TJ) integrity of polarized C2BBe1 cells cultured in Trans Well plates after exposure to five concentrations of PEGylated proteins was monitoring daily by measuring the Trans epithelial Electrical Resistance (TEER) for 3 days. Finally, the structural stability of TJ and cytoskeleton was evaluated by immune staining and colocalization of Occludin and F-actin proteins respectively, using confocal microscopy.

Results: It was observed that none of the concentrations of PEGylated proteins evaluated was able to reduce the viability of C2BBe1cells. In contrast, cell viability increased up to 124% compared with the control without treatment (100%). Likewise, it was observed that the TEER values did not have statistically significant differences between the treatments evaluated and the negative control. On the other hand, the PEGylated proteins did not produce structural changes in the TJ and cytoskeleton of polarized C2BBe1 cells. Interestingly, there was an apparent increase in the amount of occlude in that must be confirmed.

Conclusions: Anti-rotavirus PEGylated proteins obtained from the extracellular secretome of B. adolescentis do not affect the structural integrity of human intestinal C2BBe1 cells. This findings predict safety for in vivo tests.

Keywords: Bifidobacterium adolescentis, secretome proteins, Acute Diarrheal Disease, anti-rotavirus.

Biography:
Juan Carlos Ulloa Rubiano is a professor and Co-ordinator of Virology Laboratory, Faculty of Sciences of the Pontificia Universidad Javeriana, Bogotα, Colombia. Dr. Ulloa has worked in virology for 17 years, mainly with viruses that produce diarrhea (Astrovirus and Rotavirus) in infants under 5 years old. The projects in which he has been involved are related to molecular virology and antivirals development from diferent sources such as probiotics bacteria and medicinal plants. In addition to scientific publications, in 2017 he patented a bioactive antiviral fraction from a medicinal plant.

A New Software for the Prompt Identification of Infectious Diseases: Preliminary Findings

Federico Baldassi*, O. Cenciarelli, A. Malizia and P. Gaudio

University of Rome Tor Vergata, Italy

The prompt recognition of an outbreak is pivotal for the early management of a possible public health threat. Nowadays, several technologies for the rapid molecular identification of pathogens are available; together with monitoring tools (i.e. web-based surveillance tools, infectious diseases modellers and epidemic intelligence methods) – that represent important components for timely outbreak detection, these techniques contribute to the surveillance system for infectious diseases. Although these methods work well for scientific officers (i.e. Public Health specialists) and policy makers, a prompt, user-friendly and accessible tool that can support first responders, health care workers and even decision makers has not been developed yet. In this study, some preliminary findings regarding the development of a user-friendly software able to rapidly and carefully recognize a possible infectious disease are presented. This recognizing tool is being developed in the MATLAB® environment, basing the preliminary script on a regressive analysis. Because the tool has been built by integrating an infectious disease database containing at the moment 35 different agents, it will be able to be ran in an off-line mode.

Biography:
Federico Baldassi is a PhD student of Industrial Engineering department from University of Rome Tor Vergata, Italy. He is a cellular and molecular biologist and he got two master degrees in Complex Organizations Management and in Protection against CBRN events. His research interests include mathematical epidemiology, modeling of infectious diseases and Chemical Biological Radiological Nuclear (CBRN) defence.

Sensitive Virus Detection and Genotyping by Nanoparticle-based Enzyme-Amplification in a Microfluidic Device

Mengsu Yang1,2*, He Zhang1, Tao Xu1,2, Heng Zou2 and Huayang Fu1,2

1City University of Hong Kong Shenzhen Research Institute, China
2Department of Biomedical Sciences, City University of Hong Kong, China

We have developed a novel microfluidic device with microbeads array for sensitive detection and genotyping of human papillomavirus (HPV). Polystyrene microbeads were modified with primary DNA probes and electron-rich proteins and docked in the microfluidic channels in an array format. Gold nanoparticles were functionalized with horseradish peroxidase (HRP) and secondary DNA probes. Hybridization of target sequences with the DNA probes conjugated the enzyme-functionalized Au nanoparticles to the surface of microbeads, where the oxidation of biotin–tyramine by hydrogen peroxide resulted in the deposition of multiple biotin moieties onto the surface of beads, catalyzed by the immobilized electron rich proteins. Streptavidin-labeled quantum dots were then allowed to bind to the deposited biotin moieties and further amplifies the signal, leading to enhanced sensitivity with the limit of detection for specific HPV sequences at 1 fmol/L (S/R>3). The microfluidic chip-based assay was three magnitudes higher in sensitivity than that of an off-chip test. The protocol could discriminate and genotype 10 copies/L HPV genomic DNA in combination with PCR.

Biography:
Prof. Yang obtained his PhD from University of Toronto, Canada and received postdoctoral training in the Scripps Research Institute in USA. He is currently the Head of Department of Biomedical Sciences, and Yeung Kin-Man Chair Professor of Biomedical Sciences in City University of Hong Kong. The research interest of Yangʼs group focuses on the development of biochip technology and nanotechnology for molecular diagnostics and therapeutic applications. Prof. Yang has published over 280 peer-reviewed scientific papers and received 23 USA/China patents.

Understanding of Virus-Tumor Cell Interactions

Klara Valyi-Nagy*, Brian Fredericks and Tibor Valyi-Nagy

University of Illinois, College of Medicine, USA

Herpes simplex virus type 1 (HSV-1) infection of tumor cells may occur during natural viral infections of tumor patients and following the use of HSV-1-based vectors as oncolytic virotherapy agents. Three-dimensional (3D) tumor cell cultures provide a useful in vitro experimental platform to study many aspects of tumor growth and cancer therapy. During the past several years we have established a number of experimental platforms that involve inoculation of 3D melanoma cultures with HSV-1. In this presentation we will review observations made with the use of these 3D experimental platforms and demonstrate that certain melanoma cell subpopulations have increased resistance against HSV-1 infection and that HSV-1 infection may promote the growth of some melanoma cells. These findings are highly relevant to the use of HSV-1 vectors for oncolytic therapy and for a better understanding of the potential role of natural HSV-1 infections in modulating the pathogenesis of neoplasms.

Biography:
Klara Valyi-Nagy, MD is Research Associate Professor in the Department of Pathology and Associate Director of the UI Biorepository at the University of Illinois at Chicago.

Histopathologic Changes Associated with Latent Herpes Simplex Virus Type 1 Infection in Human Trigeminal Ganglia

Tibor Valyi-Nagy* and Brian Fredericks

University of Illinois, College of Medicine, USA

Herpes simplex virus type 1 (HSV-1) latent infection of human trigeminal ganglia (TG) is common and associated histopathologic changes are incompletely understood. To better understand histopathologic changes associated with HSV-1 latent infection human cadaveric TG tissues derived from six adult patients were examined for evidence of latent HSV-1 infection and associated histopathologic changes. TG tissues from three patients were found to harbor latent HSV-1 infection as demonstrated by detection of HSV-1 latency-associated transcripts by in situ hybridization and no evidence of HSV-1 protein expression by immunohistochemistry. Most TG latently infected with HSV-1 demonstrated foci of chronic inflammation. Morphometric studies indicated that the meandiameter of neuronal nuclei in latently infected TG was decreased relative to controls. These findings were similar to histopathologic changes we previously detected in a murine model of HSV-1 latent infection and provide novel information about the pathologic consequences of HSV-1 latency in the human nervous system.

Biography:
Tibor Valyi-Nagy, MD, PhD is a Professor, Director of Neuropathology and Interim Director of Anatomic Pathologyin the Department of Pathology, University of Illinois at Chicago.

H5N1-Pseudoviruses Infection In Vitro Shows Viral Interference

Anna S. Kolyasnikova1,2*, Yuliya V. Razumova1,2, Marina V. Khrapova2,4 and Tatiana V. Lipina2,3

1Novosibirsk State University, Russia
2State Scientific-Research Institute of Physiology and Basic Medicine, Russia
3Department of Pharmacology & Toxicology, University of Toronto, Canada
4RIECM FRC FTM, Russia

The current study has described features of co-infections in vitro with H5N1-pseudo lentiviruses, containing two different reporter genes: eGFP (H5N1-eGFP-plv) and mCherry (H5N1-mCherry-plv. These plvs express HA (hemagglutinin) and NA (neuraminidase), originated from influenza virus strains A/Thailand/KAN-1/04(H5N1) and A/Russia/01/2009(H1N1), respectively. Cells (MCF-7 and U87-MG) were inoculated either individually with H5N1-mCherry-plv and H5N1-eGFP-plv (mono infection) or in their combination (co-infection). The infection rates of cells after 5 days of inoculation with co-infection were lower than those with mono-infections. Furthermore co-infection occurred less frequently than predicted based on random events of infection. These data indicate the presence of viral interference when cells are infected with H5N1-pseudo typed lentiviruses at the stage of adsorption/entry of pseudo lentiviruses, into the cell.

However, when cells were infected at + 4°C (under conditions with the inhibited NA), the infection rates of co-transduced cells was near a random distribution, suggesting the important role of NA in the viral interference.

In addition, we evaluated the effect of 2 pseudo lentiviruses, containing shRNA target sequences against Phosphodiesterase 7A (PDE7A) gene, with various surface proteins (H5N1 — from influenza A virus and G — from vesicular stomatitis virus) on viability of MCF-7 cells. These pseudo lentiviruses, significantly increased cellʼ viability in a dose-dependent manner. Notably, the increment of viability was much slower in case of the H5N1-pseudo lentiviruses, These findings further support the existence of H5N1-pseudo lentiviruses, interference.

Disclaimer: This work has been supported by Russian Scientific Foundation grant 17-15-01294.

Biography:
Anna S. Kolyasnikova was graduated in Zelman Institute for the Medicine and Psychology, Novosibirsk State University, Russia.

Studies on the Interaction of Rotavirus Non Structural Proteins NSP5, NSP2 with Host Cellular Factors and their Role in Virus Replication

Varsha Narayan Tandra1* and C Durga Rao

Indian Institute of Science Bangalore, India

Rotavirus (RV) still remains the single most important etiological agent of severe diarrhea in infants and young children worldwide, with an annual mortality of over 200,000 children. It is non-enveloped, segmented, double-stranded RNA virus and belongs to the family Reoviridae. The viral genome consists of 11 segments of double-stranded RNA, which encodes 6 structural and six non-structural proteins. In infected cells, rotavirus replicates in the cytoplasm in virus induced inclusion bodies called viroplasms (VMs) which are formed by two essential viral non structural proteins NSP5 and NSP2. VMs are the sites where RNA replication and new immature double layered particle (DLP) assembly take place. Till date, the association of host cellular proteins in rotavirus with NSP5 and NSP2 has remained unexplored. By mass spectrometry analysis we have identified several host proteins showing interaction with NSP5 and NSP2, which were further confirmed by pull down assay using uninfected MA104 cell extracts. Next, we selected three host nuclear proteins hnRNP K, hnRNP D and Fibrillarin for our detailed studies and confirmed their interactions by Co-immunoprecipitation using RNase treated infected cell lysate suggesting the interaction is RNA independent. Immunofluorescence confocal microscopy revealed that these nuclear/nucleolar proteins undergo relocalization to the cytoplasm during virus infection and were sequestered into the VMs where they colocalize with NSP5 and NSP2 through direct interactions. We also observed time dependent nuclear-cytoplasmic redistribution of these proteins by western blot analysis in cytoplasmic and nuclear extracts from infected cells in comparison to that in uninfected cells. The significance of these host proteins in rotavirus infection was elucidated by siRNA-mediated knockdown and over expression in HEK293T cells. Knockdown of hnRNP D, hnRNP K and fibrillarin decreased the expression of NSP5, NSP2 and structural protein VP6 and over expression of hnRNP K and fibrillarin proteins increased the expression of viral NSP5, NSP2 and VP6.

Biography:
Varsha Narayan Tandra is a Ph.D. from Indian Institute of Science working on Molecular virology. Varsha is an enthusiastic and have keen interest in learning and gaining knowledge.

Gender Specific Medicine and Clinical Trials on Colorectal Cancer: Are Women Less Enrolled than Men or are they More Reluctant to Enlist?

Patrizia Tempia1*, Dania Brioschi2*, Francesca Crivelli3 and Francesco Leone4

1Responsible of the Hospital Psychology Unit, ASLBi, Italy
2Coordinator of the Gender Specific Medicine, Projetct, ASLBi, Italy
3Clinical Research Coordinator, Fondazione Edo ed Elvo Tempia, Italy
4Director of Oncology Department, ASLBi, Italy

Since May 30th 2019 the Gender Specific Medicine has become part of the National Health Plan (2019-2021) in Italy. This new point of view in all medical specialties stems from the growing awareness of the gender differences with the ultimate aim of ensuring the best care to each person, both male and female, further strengthening the “patient centrality” and “therapies customization” concepts. Both in international and national cancer clinical trials the stratification is not expected, then this does not allow us to measure differences in the mechanism of action, metabolism, drug effectiveness and potential adverse events in man and woman. The lower women representation compared to men is also a selection bias in itself. Among the oncological pathologies we have selected colorectal cancer because in this pathology the incidence and prevalence data are comparable in both sexes. From the review of colorectal cancer studies conducted from 2000 to 2019 at the ASLBI, Local Health Unit of Biella, the data confirm that men participate more than women in clinical trials: while 57% of the enrolled population are male, 43% female. The same inequality applies to screened and unrolled patients (58.5% over 41.5%). Thus the analysis of the international literature confirms the figure and it stimulates us to deepen the research on the reasons for this difference. Why are women less enrolled than men? Are women more reluctant to enlist? These questions suggest approaching the problem with a new perspective and analyzing both the experimenter and the enlisted subjects psychological aspects, as well as the social context in which the proposal to participate in the studies takes place. Therefore in order to ensure equal opportunities for participation in studies, the gender-specific approach becomes necessary as new evidence suggesting that the effectiveness, tolerability and toxicity of treatments are different depending on gender.

Biography:
Dr. Patrizia Tempia Valenta graduated in Clinical Psychology at the University of Turin and specialised in Clinical Psychology at Medical School. Currently she holds the position of Director of Psychological Services at the hospital of Biella (ASLBi). In 2007 she founded the Association of Volunteers “Emanuele Lomonaco- Far Pensare” and organized a literary contest “Storie di Guarigione” (Recovery stories”) on psychiatric disease, and “Contaci” a convention on Oncology and correct lifestyle. She promoted a series of plays with some artists, musicians, photographers and theatre-plays on health-care subjects. She was a teacher in several Masters in Psycho-oncology, End-life e Palliative care, and in Life-style coaching at the University of Turin and Novara. She is the author of many articles and books about Psycho-oncology, Chronicle diseases and Narrative Medicine. In particular she is co-author of “Le emozioni dei malati e dei curanti” Centro Scientifico ed. 2004; chapter “Le malattie neoplastiche e le malattie croniche ad alto impatto emotivo: La presa in carico globale” with G Ferrrandez in Psicologia e Medicina: vantaggi e prospettive- F Angeli ed 2015; she edited “Storie di Guarigione” Eventi Progetti ed. 2015; “Storie di Guarigione, dalla malattia psichiatrica grave: Una esperienza collaudata nellʼambito della medicina narrativa” in Pensieri Circolari, Narrazione, Formazione e Cura – Pensamultimedia ed. 2015; “La Psiconcologia in Musicoterapia e Oncologia” F Angeli 2015. Dania Brioschi is an experienced Healthcare Professional with a demonstrated history of working in the hospital & health care industry. She is skilled in Clinical Research, Nursing Education, Medicine, Patient Safety, and Healthcare Information Technology (HIT). She is a strong professional with an Executive Master in Management delle Aziende Sanitarie e Socio Assistenziali focused in Health care management and organisation from SDA Bocconi.

17α-Estradiol Inhibits Osteoblast Stimulation of Prostate Cancer Cell Migration and Invasion through Blockade of TGF-β1/SMAD2 Signal Pathway

Jian Shi1,2*, Brittany Duncan1, Jie Su3, Jale Manzo1, Lian Zhao1,2, He Liu4, Yuan-Shan Zhu1 and Chaoyue Zhang2

1Department of Medicine, Weill Cornell Medicine, USA
2The Third Xiangya Hospital, Xiangya School of Medicine, Central South University, China
3Cancer Biology & Genetics Program, Memorial Sloan Kettering Cancer Center, USA
4Department of Urology, Weill Cornell Medicine, USA

Prostate cancer (PC) is curable if it is diagnosed and treated in localized and regional stage. However, PC outcome is poor once it has distant metastasis. Approximately 70% to 100% of PC deaths have bone metastasis, presumably due to a specific bone microenvironment. In this study, we investigated the role of osteoblast cells in PC cell migration and invasion and revealed the effect and mechanism of 17 alpha-estradiol (αE2) on osteoblast-stimulated PC cell migration and invasion using cell culture analysis. Transwell experiments with PC and osteoblast h.FOB cell co-culture showed that PC cell migration and invasion were specifically promoted by osteoblast cells, but not cells originated from mammary gland, kidney and liver. Moreover, PC cell migration and invasion was specifically stimulated by h.FOB condition media in transwell and wound-healing assay. Multiplex immunoassays revealed that the concentration of TGF-β1 was markedly higher in the h.FOB condition media compared to other cell condition media. Treatment with TGF-β1 produced a time- and dose-dependent induction of PC cell migration and invasion and SMAD2 phosphorylation. Both the h.FOB and TGF-β1 effects were effectively suppressed by a specific TGFβ receptor inhibitor LY2157299 as well as by αE2. Most intriguing this αE2 inhibitory effect was observed at very low nanomolar concentrations and presumably mediated through estrogen receptor. These findings suggest that TGF-β1 is a major factor in mediating h.FOB cell stimulation of PC cell migration and invasion and αE2 is a potential agent to block PC cell bone metastasis, probably through inhibition of TGF-β1/SMAD2 signal pathway.

Keywords: Prostate cancer, Bone metastasis, h.FOB Cells, TGF-β/SMAD, 17α-Estradiol, Cell migration and invasion.

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