Biography:

Attila Szucs graduated in Biology and defended his PhD thesis at the University of Szeged, Faculty of Science. His area of research is in the analysis of high-throughput DNA/RNA data. His focus is on characterization of mRNAs using both quantitative and qualitative methods and combining data from various source into databases. He has published 36 papers on these subjects in reputed journals up to now.

Abstract:

The Oxford Nanopore Technologies MinION and the Pacific Biosciences Sequel, RSII long-read sequencing platforms are capable to sequence full length transcripts, although several limitations are still occurred. The sequencing depths are much lower compared to short-read sequencing methods. One of the main problem is to distinguish between the degraded mRNAs molecules and full-length transcripts. This problem can be relatively easily solved for the abundant genes, but it is difficult for low abundant ones. Furthermore, not a trivial task to distinguish between the sequences which conatin false and true intronic sites. The false introns are generated by the strand switching  effect of the reverse transcriptase. False prime sites in the genome are another source of  the sequencing error. The currently available mapping softwares not always provide the most significant matches. To circumvent the above mentioned problems, we have written several routines. These routine scripts are able to correct the alignments at the end of mapped reads and remove extra false exons. Our scripts add extra parameters to the BAM files, which contain information about the positions and similarity of adaptor sequence. These routins use statistic methods to distinguish between the real and the false transcription start and end sites (TSS and TES). The program removes the potential TSS and TES sites derived from false priming. Moreover, the reads which contain false introns are also removed by our routines. Finally, our programs are able to collect the “real” transcripts and their abundance.   

Biography:

Antonio Alberto Rodríguez Sousa has an extensive experience in systems dynamics and modeling. With biological training, he develops part of his personal work at Complutense University of Madrid on the simulation of the effects presented by different treatments on alveolar development in people with cystic fibrosis, a disease in which he has a high degree of specialization on the lung involvement in adults. In this sense, he has evaluated how the application of different drugs contributes to improve the hope and quality of life of patients. Specifically, and under the appropriate assumptions, it tries to make the scientific and medical sector aware that the application of a gene therapy using a virus as a transmission vector would contribute to a reversion of cystic fibrosis when the affected cellular processes are resumed. Finally, research in this area is essential to be able to propose curative treatments for diseases of genetic transmission.

Abstract:

Statement of the Problem: Cystic fibrosis is the autosomal recessive genetic disorder with the highest incidence in the Caucasian population, being caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Although the current treatments focus on increasing the quality of life of the patient, a gene therapy of viral origin that could involve the regeneration of damaged alveolar parenchyma is located in the experimental phase. The purpose of the present work is to establish a theoretical framework of the disease from which clinical work can be carried out aimed at the development of new therapeutic techniques.

Methodology & Theoretical Orientation: A dynamic simulation model was developed on patients' hope and quality of life, taking the number of pulmonary alveoli as an estimator of respiratory function. The model was calibrated in face of different clinical situations: healthy individual; untreated sick individual; sick individual with conventional treatments; and sick individual treated with gene therapy.

Findings: The study showed that the life expectancy of sick individuals was significantly reduced when compared with healthy individuals. On the other hand, while the application of conventional treatments reflected an improvement in the quality of life of the simulated patients, the administration of a viral-type gene therapy was consolidated as an option in which the treated individual’s hope of life was not affected due to the disease.

Conclusion & Significance: By using viruses as a biotechnological tool, being transmission vectors of functional copies of the CFTR gene that would be inserted into the host's DNA, it would be possible to correctly resume the cellular processes altered by the disease. Therefore, the research in virology and its application in gene therapies are essential to develop curative treatments that suppose a new clinical horizon.

Biography:

Abstract:

Background: Before the era of combination therapy (c-ART) more than 90% of the patients with HIV-infection died of one or more opportunistic infections (OI). We and others noted early on that cytomegalovirus (CMV) was an important pathogen in these patients. We investigated: all OIs and opportunistic cancers (OCs) in patients who died with CD4+ counts below 100x106/mL; the prevalence of CMV encephalitis (CMV-E) and Korsakoff syndrome; CMV retinitis (CMV-R) in relation to CMV-E; CMV adrenalitis (CMV-A) and its relation to CMV-R and CMV-E; the correlation between CMV disease and other OIs and OC and; the interaction between CMV, Epstein Barr virus (EBV) and other human herpes viruses in a case of anaplastic large cell lymphoma (ALCL).

Material & Methods: We followed all patients died at Venhälsan from 1989-1996, with intensive blood testing, X-rays, CT scans, Synacthen tests, neurological examinations, and ophthalmologic examinations to.

Results: Of all 219 patients died with CD4+ < 100x106/mL, 87% showed signs of reactivated CMV-infection. CMV-R was found in 84, CMV-E in 65, CMV-A in 41 and CMV in the gastrointestinal tract in 21. Mycobacterial infection was found in 87 and toxoplasmosis in 29. Kaposi’s sarcoma was the most common tumor (68 cases) followed by 22 patients with malignant lymphoma and 20 with CNS-lymphoma. CMV-reactivation was seen in most. A case of primary CMV-infection leading to a malignant lymphoma by interaction with two other herpes viruses (EBV and HHV-8) was also seen.

Conclusion: CMV-infection was the main OI in AIDS-patients during the pre-c-ART era and the main cause of death by itself or together with other OIs. Reactivation of CMV was found in 87%. The most important CMV manifestations were CMV-R, CMV-E and CMV-A that seemed to occur at the same time. This is still today important in patients with CD4+ <100x106/mL without access to modern HIV-treatment. These findings reveal the intimate interaction between HIV and CMV which should be considered in all co-infected patients also today.

Biography:

Abstract:

AIDS is an ever-growing public health concern in China. Some HIV-infected patients get admitted because of severe opportunistic infections (OIs) which are the significant complication of HIV infection. By the end of 2014, there were 501,000 reported cases of people living with HIV/AIDS across China, with 66,035 people in Sichuan province alone. However, the prevalence and spectrum of OIs among Chinese HIV-infected patients are poorly understood. In the present study, 2298 cases of HIV infection in Sichuan were retrospectively investigated at the Public Health Clinical Center of Chengdu. We found that bacterial pneumonia (25.8%) was the most common OIs, followed by candida (18.3%), PCP (11.9%), tuberculosis (11.5%), infectious diarrhoea (9.3%), cryptococcus (7.3%), CMV (4.9%), toxoplasmosis (4.6%), HCV (4.0%), NTM (2.2%), and PM (0.3%). A noteworthy observation in this study is that CD4+ T cell count was found to be a predictor of some OIs. The specific pathogens causing bacterial pneumonia and/or candida infections and an effect of TB on CD4+ T cell count were also analysed between HIV-infected and non-HIV-infected patients. Understanding the spectrum of OIs in Sichuan could help us develop successful and efficient public health strategies. Such information could also help clinicians diagnose and initiate proper treatment more rapidly in hard-hit areas with limited resources.

Biography:

Abstract:

Noroviruses (NoVs) are major causal agents of acute gastroenteritis in humans. NoV GII.4 is the predominant genotype globally. However, uncommon and minor types of NoVs are consistently detected and some have been shown to dominate over GII.4. Therefore, the prevalence of dominant and uncommon NoVs makes the identification of these viruses important for the prediction and prevention of pandemics. In this study, the full-genome sequence of a NoV (strain JW) detected in Korea was extensively characterized. The full-length genome was 7510 nucleotides long, and phylogenetic analysis based on the whole-genome sequences, including open reading frame (ORF)1, ORF2, and ORF3, indicated that it belonged to the GII.21 genotype. Strain JW showed maximum identity with strain YO284; however, comparison of the amino acid sequence of ORF2, which functions as an antigen, showed substitutions in several amino acids. GII.21 is not a prevalent epidemiological agent of acute gastroenteritis in humans, but it is consistently found in gastroenteritis patients from several countries. The present study provides the first full-genome sequence analysis of NoV GII.21 isolated from a patient in Korea. Our findings provide not only valuable genome information but also data for epidemiology studies, epidemic prevention, and vaccine development strategies.

Biography:

Remziye Nalcacioglu has completed her PhD in 2003 from Karadeniz Technical University, in Turkey. She has been working in the same universirty as an academician. She is working on insect viruses including baculoviruses and iridoviruses.

Abstract:

The genome of Chilo iridescent virus (CIV) has two open reading frames (ORFs) with matrix metalloproteinase (MMP) domains. The proteins encoded by 136R and 165R ORFs contain 178 amino acids with over 40% amino acid sequence identity to hypothetical metallopeptidases of other viruses and 264 amino acids with over 40% amino acid sequence identity to metallopeptidases of a large group of organisms including primarily variety of Drosophila species, respectively. These proteins possess conserved zinc-binding motifs in their catalytic domains. In this study, we focused on the functional analysis of these ORFs. These ORFs were cloned into the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) Bac-to-Bac baculovirus expression-vector system, expressed in insect Sf9 cells with an N-terminal His tag and purified after 96 hours post infection to homogeneity by using Ni-NTA affinity chromatography. Western blot analyses of purified 136R and 165R proteins with histidine tags resulted in 24 and 34 kDa protein bands, respectively. Biochemical assays with the purified proteins, performed using dye-impregnated collagen (Azocoll) and Azo-casein as substrates, showed that both proteins have protease activity. The enzymatic activities were inhibited by metalloproteinase inhibitor EDTA. Effects of these proteins were also investigated on Galleria mellonella larvae. Insecticidal activities were carried out by injecting the larvae with the AcMNPV carrying 136R and 165R ORFs. Results showed that the baculoviruses harboring the iridoviral metallopeptidases caused early death of the larvae compared to control group which was performed with only wild type AcMNPV.  All these data suggest that the CIV 136R and 165R ORFs encode functional metalloproteinases which can be utilized in biological control of lepidopteran pests.

Biography:

Larry Martínez is a doctor specialized in internal medicine and infectious diseases, with experience in the management of vulnerable population since he was a medical student as a volunteer of the Colombian Red Cross. He is currently number member of de Infectology Colombian Asociation, working in third level hospitals ans clinics in the city of Medellín of Colombia through the HIV / AIDS and Tuberculosis program. Focused on advancing studies and observations regarding the incidence of viral agents and other groups of opportunists in HIV / AIDS patients. It aims to demonstrate the real incidence and prevalence of some germs and their role as coinfectants in patients with HIV / AIDS.

Abstract:

Parvovirus B19 is a global infection that can cause serious and life threatening disorders in susceptible patient groups. [1]  Viruses of the family Parvoviridae, are among the smallest viruses described. The first pathogenic human parvovirus was discovered and named B19 from the coding of a serum sample, number 19 in panel B, that gave anomalous results during testing for hepatitis B. [2]. Parvovirus B19 genotype 1, has a worldwide distribution. Genotypes 2 and 3 tend to be found in Europe and Africa.[3]. We present the clinical and epidemiological description of cases of aplasia of the red series without affecting hematimetric indices in HIV positive patients with positive serology to Parvovirus B19 admitted to hospitalization between April 2016 to April 2017. Sixteen cases were documented, 11 men (69%) and 5 women (31%) with an average age of 40.7 and 44.4 years respectively, 5 patients (31.25%) had positive IgM levels without IgG activity documented in the same sample and 4 of these 5 patients had abandoned treatment for their HIV (80%), the average in grams per deciliter of hemoglobin and hematocrit at the time of sampling was 8.92 g / dl  and 28.6 g / dl respectively. All the patients included had IgG titles but only 6 had positive titles with a positive reference value> 11 (37.5%) and an average CD4 / ul cells count of 115 for men and 187.2 for women. Draws attention the most prevalent opportunist in the sampling is mycobacterium tuberculosis. High viremia could represent a great risk in plasma derivatives, [4-6] and all our patients required transfusions of red blood cells units (1.4 and 4.7 times, for women and men respectively). In patients with a clearly disturbed immunity, its relevant a deep molecular investigation to define real implications, epidemiology and distribution of this agent in our country

Biography:

Salik Nazki is currently pursuing PhD from Chonbuk National University, South Korea. He has completed DVM and Master’s in Veterinary Microbiology and Immunology from SKUAST-Kashmir, India. The major focus of his research is on PRRSV and is well versed with the virological and immunological techniques. Previously, during his Master’s degree, his research work was based on anaerobic bacteria and also has experience on other viruses. He has recently published some research articles in reputed journals

Abstract:

Porcine reproductive and respiratory syndrome (PRRS) virus causes significant economic losses to swine industries. To develop an effective vaccine against PRRSV is a major challenge due to huge genetic variation in PRRSV isolates. However, modified live virus (MLV) vaccines are widely used for homologous protection though there have been safety issues as vaccine viruses reverted to virulent. Besides, the existence of both genotypes together in field facilitated to emerge new strains. Therefore, regular characterization of prevalent isolates is essential to enhance protective-efficacy against PRRSV isolates. In the present study, seven type 2 PRRSV strains (NA4, NA8, NA10, NA31, NA45, NA73 and 10D415) prevalent in Korea were characterized for their immunomodulatory effects and pathogenicity in pigs. Eight piglets were infected with each strain including VR2332 (prototype of type 2 PRRSV) and kept upto 28 dpi. Serum viremia and body weight were measured weekly. Pigs from each group were euthanized at 14 and 28 dpi. Pathological evaluation was conducted and various samples were collected. Various T cell responses were analyzed in peripheral blood mononuclear cells (PBMCs) and tissues collected from the pigs. In results, NA10 and 10D415 showed highest levels of virulence as they induced high body temperature, reduced weight gain and showed high mortality in challenged pigs though 10D415 induced lower viral loads in serum and nasal fluids. On the other hand, NA8 which shares highest sequence homology with VR2332 showed lowest virulence. Similarly, the viruses evaluated in the current study showed various levels of γδ-T, Th1, Th17 and CTL responses in PBMCs or tissues. In conclusion, the present study suggested that type 2 PRRSVs in Korea showed a wide range of pathogenicity and immune responses and the information might be helpful in designing efficient vaccine platform against PRRSV infection.

Biography:

Chang Gi Jeong has completed his MS in Veterinary Medicine (Microbiology) from Chonbuk National University (CBNU), South Korea. During his Master’s course, he worked on the Clostridium novyi and studied the genome characteristics of isolates using Next Generation Sequencing (NGS). Currently, he is pursuing PhD course at the same university. His major research focuses on PRRSV and Japanese encephalitis virus. He is well-adapted in animal studies mostly dealing with farm animals.

Abstract:

Porcine reproductive and respiratory syndrome (PRRS) is a challenging threat to the swine industries caused by PRRS virus and characterized by reproductive failures in pregnant and respiratory distress in piglets. As an RNA virus, PRRSV mutated quickly and evolved continuously, which caused huge genetic and antigenic variation within genotypes or even in the same viruses. Furthermore, the biological properties of PRRSV responsible for viral pathogenesis and host-immune responses have not been characterized clearly. Many previous studies have been conducted by using a respiratory disease model in weaned piglets for convenience but the reproductive disease caused by PRRSV is still poorly understood. Therefore, the present study was aimed to demonstrate the pathogenicity and immune response following infections with both type 1 (D40 and CBNU0495) and type 2 (K07-2273 and K08-1054) PRRSV strains in pregnant sows. Two pregnant sows were infected with each virus at 93 days of gestation. At 21 days after infection, all sows and their fetuses were euthanized for pathological evaluation. In addition, viral loads were measured in serum samples from the sows and various tissues collected from the sows and their fetuses and fetal weights were recorded. In results, CBNU-0495 and K08-1054 showed higher virulence as compared with other viruses as they exhibited higher levels of viral loads in sera and tissues of sows and fetuses and also caused higher levels of weight losses in fetuses.

Biography:

Abstract:

The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is an insect virus belonging to the Baculoviridae family. The 134 kbp long double-stranded circular DNA of the virus encompasses 150 tightly packed open reading frames. Genes of the AcMNPV are expressed in three phases: early, late and very late. Promoters of early genes are recognized by the host RNA polymerase, and usually consist of a canonical TATA motif located upstream of the transcriptional start site. At the same time some early transcripts start from the arthropod initiator element (CAGT). Late and very late genes are transcribed by the viral RNA polymerase, which recognizes a late initiator sequence (TAAG). A short-read sequencing technique was used previously to elucidate the structure of the AcMNPV transcripts; however short-read sequencing cannot tackle the highly complex, overlapping nature of the viral transcriptome. In this study we found and annotated four novel putative protein coding, four non-coding transcripts and forty-seven novel length isoforms of previously annotated transcripts using the Oxford Nanopore Technologies’ MinION and the Pacific Biosciences Sequel platforms. We demonstrated that the canonical promoters and initiators of a transcript are in concordance with the motifs found upstream of its isoforms. We also discovered the extensive overlapping nature of the viral transcriptome. Additionally we determined the expression characteristics of novel transcript isoforms and revised it for the already annotated transcripts using long-read real-time sequencing of the AcMNPV transcriptome.

Biography:

Remziye Nalçacioglu has completed her PhD in 2003 from Karadeniz Technical University, in Turkey. She has been working in the same universirty as an academician. She is working on insect viruses including baculoviruses, iridoviruses and Bacillus thuringiensis bacteria.

Abstract:

A novel group of insecticidal proteins named vegetative insecticidal proteins (Vıp) are produced by Bacillus thuringiensis (Bt) bacteria during its vegetative stage. In this study we characterised the vip3 genes of two local Bt isolates (BnBt, MnD). Firstly, partial-purified Vip3 proteins of some local Bt isolates were tested against the Spodoptera littoralis larvae. After obtaining good insecticidal activity with BnBt and MnD isolates, Vip3 proteins of these bacteria were purified from supernatants of bacterial cultures by ion exchange chromatography. Purified proteins were subjected to SDS-PAGE analysis and 90 kDa band of proteins were determined. These purified proteins were tested against S. littoralis larvae. Results showed that Vip3 proteins of BnBt and MnD produced 86.66% and 83.33% insecticidal activity on S. littoralis larvae, respectively. The lethal concentrations (LC₅₀) of BnBt and MnD were determined as 41.860 ng and 55.154 ng, respectively. These results suggest that vip3 genes of our local isolates may be alternatives for preventing resistance in various insect–pest species. Also these proteins may be used at developing bio-pesticides.

Biography:

Abstract:

According to results another researches in this study mesial temporal lobe epilepsy (MTLE) is shown to be associated with human herpes virus 6 and 7 types (HHV6/HHV7) neuroinfections. We also demonstrate that the epileptic process is associated with an systemic inflammatory reaction, and that the proinflammatory cytokine, the tumor necrosis factor-alpha (TNF-α), is able to potentiate the reproduction of the herpes viruses. The study group (SG) included 43 patients between 16 and 60 years with MTLE and HHV neuroinfections, diagnosed according to the PCR of the cerebrospinal fluid (CSF), serum or abnormal serum/CSF IgG ratio. The control group (CG) included 30 patients of similar age with MTLE, but without the HHV neuroinfections. The concentration of TNF-α in the serum was determined by enzyme-linked immunosorbent assay ("VektorBEST" RF; N=0-50 pg/ml). Patients of the SG had high concentrations of TNF-α in serum (288±44.7 pg/ml), that were significantly higher than in the CG (p<0.05; Z<Z_0.05). Serum concentrations of TNF-α greater than 100 pg/ml were associated with the severe general condition of the patients, more severe epileptic syndrome, a long history of illness, deep organic brain damage, low sensitivity to anticonvulsant drugs, overall with a poor prognosis. In patients with MTLE and HHV6/HHV7 neuroinfections marked systemic inflammatory response syndrome was noted, which affected the severity of the symptoms in the patient. TNF-α, therefore, can be used as a biomarker for an objective assessment of the severity and prognosis of the disease in patients with MTLE induced by HHV6/HHV7.

Biography:

Izabela Serafinska is a PhD candidate in the Division of the Veterinary Medicine in Warsaw University of Life Sciences, Poland. Her main research topic is autophagy in neurons during EHV-1 infection. Her scientific interests include neuroinfections and microscopy.

Abstract:

Autophagy is an evolutionary conservative, intracellular process. It plays an important role in maintaining homeostasis in cell through degradation of their own components such as proteins or organelles via lysosomes. Viral infection can modulate this process. During infection autophagy may be stimulated or inhibited, because virions can be degraded in autophagolysosomes. Some viruses can utilize this process for replication. To investigate the influence of this process on the level of viral replication, stimulators or inhibitors of autophagy can be applied. Primary cultures of murine neurons were treated with autophagy inductors (rapamycin and temozolomide) and inhibitors (wortmannin and chloroquine) for 24 h. After incubation neurons were infected with equine herpesvirus type 1 (EHV-1) for 2, 24 and 48 h. In experiments non-neuropathogenic Jan-E EHV-1 strain was used. Level of viral replication was analyzed using real-time PCR. Presence and localization of viral antigens and LC3B protein (autophagy marker) were detected using confocal microscopy. Results obtained from real-time PCR showed an increase of level of viral DNA in control. At 2 h p.i., after chemical treatment, increase in EHV-1 DNA amount was observed. At 24 h p.i. this level was decreased and then at 48 h p.i. increased again, except sample treated with rapamycin. In this cells level of viral DNA also decreased at 48 h p.i. Immunofluorescence staining showed that LC3B protein was presented inside neurons during EHV-1 infections. In neurons at 2 and 48 h p.i. higher number of viral particles was observed in comparison to 24 h p.i. Results showed changes in replication kinetics during EHV-1 infection after using autophagy modulators which can be useful in antiviral therapy.