Search results for: human herpes virus
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 8491

Search results for: human herpes virus

8491 Inhibition of Mixed Infection Caused by Human Immunodeficiency Virus and Herpes Virus by Fullerene Compound

Authors: Dmitry Nosik, Nickolay Nosik, Elli Kaplina, Olga Lobach, Marina Chataeva, Lev Rasnetsov

Abstract:

Background and aims: Human Immunodeficiency Virus (HIV) infection is very often associated with Herpes Simplex Virus (HSV) infection but HIV patients are treated with a cocktail of antiretroviral drugs which are toxic. The use of an antiviral drug which will be active against both viruses like ferrovir found in our previous studies is rather actual. Earlier we had shown that Fullerene poly-amino capronic acid (FPACA) was active in case of monoinfection of HIV-1 or HSV-1. The aim of the study was to analyze the efficiency of FPACA against mixed infection of HIV and HSV. Methods: The peripheral blood lymphocytes, CEM, MT-4 cells were simultaneously infected with HIV-1 and HSV-1. FPACA was added 1 hour before infection. Cells viability was detected by MTT assay, virus antigens detected by ELISA, syncytium formation detected by microscopy. The different multiplicity of HIV-1/HSV-1 ratio was used. Results: The double viral HIV-1/HSV-1 infection was more cytopathic comparing with monoinfections. In mixed infection by the HIV-1/HSV-1 concentration of HIV-1 antigens and syncytium formations increased by 1,7 to 2,3 times in different cells in comparison with the culture infected with HIV-1 alone. The concentration of HSV-1 increased by 1,5-1,7 times, respectively. Administration of FPACA (1 microg/ml) protected cells: HIV-1/HSV-1 (1:1) – 80,1%; HIV-1/HSV-1 (1:4) – 57,2%; HIV-1/HSV-1 (1:8) – 46,3 %; HIV-1/HSV-1 (1:16) – 17,0%. Virus’s antigen levels were also reduced. Syncytium formation was totally inhibited in all cases of mixed infection. Conclusion: FPACA showed antiviral activity in case of mixed viral infection induced by Human Immunodeficiency Virus and Herpes Simplex Virus. The effect of viral inhibition increased with the multiplicity of HIV-1 in the inoculum. The mechanism of FPACA action is connected with the blocking of the virus particles adsorption to the cells and it could be suggested that it can have an antiviral activity against some other viruses too. Now FPACA could be considered as a potential drug for treatment of HIV disease complicated with opportunistic herpes viral infection.

Keywords: antiviral drug, human immunodeficiency virus (hiv), herpes simplex virus (hsv), mixed viral infection

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8490 Isolation and Elimination of Latent and Productive Herpes Simplex Virus from the Sacral and Trigeminal Ganglions

Authors: Bernard L. Middleton, Susan P. Cosgrove

Abstract:

There is an immediate need for alternative anti-herpetic treatment options effective for both primary infections and reoccurring reactivations of herpes simplex virus types 1 (HSV-1) and 2 (HSV-2). Alternatives currently approved for the purposes of clinical administration includes antivirals and a reduced set of nucleoside analogues. The present article tests a treatment based on a systemic understanding of how the herpes virus affects cell inhibition and breakdown and targets different phases of the viral cycle, including the entry stage, reproductive cross mutation, and cell-to-cell infection. The treatment consisted of five immunotherapeutic core compounds (5CC), which were hypothesized to be capable of neutralizing human monoclonal antibodies. The tested 5CC were noted as being functional in the application of eliminating the DNA synthesis of herpes viral interferon (IFN) - induced cellular antiviral response. They were here found to neutralize antiviral reproduction by blocking cell-to-cell infection. The activity of the 5CC was tested on RC-37 in vitro using an assay plaque reduction and in vivo against HSV-1 and HSV-2. The 50% inhibitory concentration (IC50) of 5CC was 0.0009% for HSV-1 plaque formation and 0.0008% for HSV-2 plaque formation. Further tests were performed to evaluate the susceptibility of HSV-1 and HSV-2 to anti-herpetic drugs in Vero cells after virus entry. There were high-level markers of the 5CC virucidal activity in the viral suspension of HSV-1 and HSV-2. These concentrations of the 5CC are nontoxic and reduced plaque formation by 98.2% for HSV-1 and 93.0% for HSV-2. Virus HSV-1 and HSV-2 titers were reduced significantly by 5CC to the point of being negative, ranging 0.01–0.09 in 72%. The results concluded the 5CC as being an effective treatment option for the herpes simplex virus.

Keywords: synergy pharmaceuticals, herpes treatment, herpes cure, synergy pharmaceuticals treatment

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8489 Mathematical Model of the Spread of Herpes Simplex Virus Type-2 in Heterosexual Relations with and without Condom Usage in a College Population

Authors: Jacob A. Braun

Abstract:

This paper uses mathematical modeling to show the spread of Herpes Simplex type-2 with and without the usage of condoms in a college population. The model uses four differential equations to calculate the data for the simulation. The dt increment used is one week. It also runs based on a fixated period. The period chosen was five years to represent time spent in college. The average age of the individual is 21, once again to represent the age of someone in college. In the total population, there are almost two times as many women who have Herpes Simplex Type-2 as men. Additionally, Herpes Simplex Type-2 does not have a known cure. The goal of the model is to show how condom usage affects women’s chances of receiving the virus in the hope of being able to reduce the number of women infected. In the end, the model demonstrates that condoms offer significant protection to women from the virus. Since fewer women are infected with the virus when condoms are used, in turn, fewer males are infected. Since Herpes Simplex Type-2 affects the carrier for their whole life, a small decrease of infections could lead to large ramifications over time. Specifically, a small decrease of infections at a young age, such as college, could have a very big effect on the long-term number of people infected with the virus.

Keywords: college, condom, Herpes, mathematical modelling

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8488 Anterior Uveitis Caused by Infection with Cytomegalovirus and Herpes Simplex Virus Type I at Cicendo Eye Hospital Bandung

Authors: Shinta Stri Ayuda Nur Setyaningsih

Abstract:

Anterior uveitis is often triggered by viral infections such as herpes simplex virus (HSV) and cytomegalovirus (CMV). This study aims to provide an overview of the demographic and clinical characteristics of patients with anterior uveitis caused by CMV and HSV infection at PMN Cicendo Eye Hospital Bandung. This study used a retrospective observational method. Data were collected from the medical records of patients who visited the PMN Infection and Immunology Polyclinic at Cicendo Eye Hospital between February and July 2023. The results showed that anterior uveitis associated with HSV and CMV viruses often occurs in the elderly and more in women. The most common clinical symptoms are red eyes and decreased visual acuity, with a gradual onset of symptoms. Complications that often arise are cataracts and glaucoma. This study provides a deeper understanding of anterior uveitis caused by infection with HSV and CMV viruses.

Keywords: uveitis anterior, cytomegavirus, herpes simplex virus type I ELISA

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8487 In vitro Antiviral Activity of Ocimum sanctum against Animal Viruses

Authors: Anjana Goel, Ashok Kumar Bhatia

Abstract:

Ocimum sanctum, a well known medicinal plant is used for various alignments in Ayurvedic medicines. It was found to be effective in treating the humans suffering from different viral infections like chicken pox, small pox, measles and influenza. In addition, curative effect of the plant in malignant patients was also reported. In the present study, leaves of this plant were screened against animal viruses i.e. Bovine Herpes Virus-type-1 (BHV-1), Foot and Mouth disease virus (FMDV) and Newcastle Disease Virus (NDV). BHV-1 and FMDV were screened in MDBK and BHK cell lines respectively using cytopathic inhibition test. While NDV was propagated in chick embryo fibroblast culture and tested by haemagglutination inhibition test. Maximum non toxic dose of aqueous extract of Ocimum sanctum leaves was calculated by MTT assay in all the cell cultures and nontoxic doses were used for antiviral activity against viruses. 98.4% and 85.3% protection were recorded against NDV and BHV-1 respectively. However, Ocimum sanctum extract failed to show any inhibitory effect on the cytopathic effect caused by FMD virus. It can be concluded that Ocimum sanctum is a very effective remedy for curing viral infections in animals also.

Keywords: bovine herpes virus-type-1, foot and mouth disease virus, newcastle disease virus, Ocimum sanctum

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8486 Diverse Sensitivity to Ultraviolet Radiation of DNA and RNA Viruses

Authors: Nickolay Nosik, Dmitry Nosik, Marina Bochkova, Nina Kondrashina, Olga Lobach

Abstract:

The bactericidal effect of UV radiation is known for long time and widely used for inactivation of pathogens but for viruses it is not so uniform. Due to a wide variety of viruses their sensitivity to UV radiation is quite different and not quite predictable. The goal of the study was to determine the inactivation kinetics of UV radiation ( 254 nm) of the viruses of social importance (HIV), as well as test-viruses (poliovirus, adenovirus) used for the evaluation of the viral inactivation efficacy of germicides. Methods: DNA viruses- adenovirus, type 5; Herpes simplex virus (HSV), type 1, and RNA viruses–human immunodeficiency virus (HIV), type 1 and poliovirus, type 1 (Sabin strain) were obtained from State collection of viruses ( The D.I. Ivanovsky Institute of Virology). The source of UV radiation was a 15-watt low-pressure mercury vapor lamp (over 60% 254nm). The samples of 5cm2 were placed direct under the UV lamp flow (h-0.3m). Log reduction value was used as a marker for the rate of virus inactivation. Results: The data obtained indicate that poliovirus (one of the viruses most resistant to chemical germicides) and HSV are rather sensitive to UV radiation ( D90 =250-311 J/m2). Adenovirus is much more resistant to UV radiation (750 J/m2 ). The kinetics of adenovirus inactivation : 0 min- 5.0 lg TCID50, 10 min - 5,0, 15 min -4,0, 30 min – 3.5, 60 min – 1,0, 75 min -0,5 lg TCID50, 90 min –virus not detectable. HIV is most resistant to UV radiation among the studied viruses. It takes more than 4 hrs to inactivate the virus on the surface. D90 = 2000 J/m2 Conclusion: The results of the study show that there is no direct dependence between sensitivity to UV light and the size of the virion or presence\absence of the envelope of the virus. Poliovirus and adenovirus are small viruses (20-30nm poliovirus and 70-90nm adenovirus) and both are non-enveloped viruses but adenovirus 3-fold more resistant to UV radiation than poliovirus. It can be expected that viruses with more complicate structure, like Herpes virus (200nm) or HIV (80-100 nm), would be more sensitive to UV light. However, the very high resistance of HIV to UV radiation needs further investigation. The diverse resistance of the different viruses to UV radiation should be taken into the account when UV light is used to inactivate infectious viruses in hospitals and other public environments.

Keywords: HIV, HSV, inhibition of viruses, UV radiation

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8485 Medicinal Plants: An Antiviral Depository with Complex Mode of Action

Authors: Daniel Todorov, Anton Hinkov, Petya Angelova, Kalina Shishkova, Venelin Tsvetkov, Stoyan Shishkov

Abstract:

Human herpes viruses (HHV) are ubiquitous pathogens with a pandemic spread across the globe. HHV type 1 is the main causative agent of cold sores and fever blisters around the mouth and on the face, whereas HHV type 2 is generally responsible for genital herpes outbreaks. The treatment of both viruses is more or less successful with antivirals from the nucleoside analogues group. Their wide application increasingly leads to the emergence of resistant mutants In the past, medicinal plants have been used to treat a number of infectious and non-infectious diseases. Their diversity and ability to produce the vast variety of secondary metabolites according to the characteristics of the environment give them the potential to help us in our warfare with viral infections. The variable chemical characteristics and complex composition is an advantage in the treatment of herpes since the emergence of resistant mutants is significantly complicated. The screening process is difficult due to the lack of standardization. That is why it is especially important to follow the mechanism of antiviral action of plants. On the one hand, it may be expected to interact with its compounds, resulting in enhanced antiviral effects, and the most appropriate environmental conditions can be chosen to maximize the amount of active secondary metabolites. During our study, we followed the activity of various plant extracts on the viral replication cycle as well as their effect on the extracellular virion. We obtained our results following the logical sequence of the experimental settings - determining the cytotoxicity of the extracts, evaluating the overall effect on viral replication and extracellular virion.During our research, we have screened a variety of plant extracts for their antiviral activity against both virus replication and the virion itself. We investigated the effect of the extracts on the individual stages of the viral replication cycle - viral adsorption, penetration and the effect on replication depending on the time of addition. If there are positive results in the later experiments, we had studied the activity over viral adsorption, penetration and the effect of replication according to the time of addition. Our results indicate that some of the extracts from the Lamium album have several targets. The first stages of the viral life cycle are most affected. Several of our active antiviral agents have shown an effect on extracellular virion and adsorption and penetration processes. Our research over the last decade has shown several curative antiviral plants - some of which are from the Lamiacea family. The rich set of active ingredients of the plants in this family makes them a good source of antiviral preparation.

Keywords: human herpes virus, antiviral activity, Lamium album, Nepeta nuda

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8484 Human Endogenous Retrovirus Link With Multiple Sclerosis Disease Progression

Authors: Sina Mahdavi

Abstract:

Background and Objective: Multiple sclerosis (MS) is an inflammatory autoimmune disease of the CNS that affects the myelination process in the central nervous system (CNS). Complex interactions of various "environmental or infectious" factors may act as triggers in autoimmunity and disease progression. The association between viral infections, especially human endogenous retrovirus (HERV) and MS is one potential cause that is not well understood. This study aims to summarize the available data on HERV infection in MS disease progression. Materials and Methods: For this study, the keywords "Multiple sclerosis", "Human endogenous retrovirus", and "central nervous system" in the databases PubMed, Google Scholar, Sid, and MagIran between 2016 and 2022 were searched and 14 articles chosen, studied, and analyzed. Results: In the leptomeningeal cells of MS patients, a retrovirus-like element associated with reverse transcriptase (RT) activity called multiple sclerosis-associated retroviruses (MSRV) has been identified. HERVs are expressed in the human CNS despite mechanisms to suppress their expression. External factors, especially viral infections such as influenza virus, Epstein-Barr virus, and herpes simplex virus type 1, can activate HERV gene expression. The MSRV coat protein is activated by activating TLR4 at the brain surface, particularly in oligodendroglial progenitor cells and macrophages, leading to immune cascades followed by the downregulation of myelin protein expression. The HERV-K18 envelope gene (env) acts as a superantigen and induces inflammatory responses in patients with MS. Conclusion: There is a high expression of endogenous retroviruses during the course of MS, which indicates the relationship between HERV and MS, that this virus can play a role in the development of MS by creating an inflammatory state. Therefore, measures to modulate the expression of endogenous retroviruses may be effective in reducing inflammatory processes in demyelinated areas of MS patients.

Keywords: multiple sclerosis, human endogenous retrovirus, central nervous system, MSRV

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8483 Detection of Elephant Endotheliotropic Herpes Virus in a Wild Asian Elephant Calf in Thailand by Using Real-Time PCR

Authors: Bopit Puyati, Anchittha Kaewchana, Nuntita Ruksachat

Abstract:

In January 2018, a male wild elephant, approximately 2 years old, was found dead in Phu Luang Wildlife Sanctuary, Loei province. The elephant was likely to die around 2 weeks earlier. The carcass was decayed without any signs of attack or bullet. No organs were removed. A deadly viral disease was suspected. Different organs including lung, liver, intestine and tongue were collected and submitted to the veterinary research and development center, Surin province for viral detection. The samples were then examined with real-time PCR for detecting U41 Major DNA binding protein (MDBP) gene and with conventional PCR for the presence of specific polymerase gene. We used tumor necrosis factor (TNF) gene as the internal control. In our real-time PCR, elephant endotheliotropic herpesvirus (EEHV) was recovered from lung, liver, and tongue whereas only tongue provided a positive result in the conventional PCR. All samples were positive with TNF gene detection. To our knowledge, this is the first report of EEHV detection in wild elephant in Thailand. EEHV surveillance in this wild population is strongly suggested. Linkage between EEHV in wild and domestic elephants should be further explored.

Keywords: elephant endotheliotropic herpes virus, PCR, Thailand, wild Asian elephant

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8482 The Link of the Human Immunodeficiency Virus With the Progression of Multiple Sclerosis Disease

Authors: Sina Mahdavi

Abstract:

Multiple sclerosis (MS) is a progressive inflammatory autoimmune disease of the CNS that affects the myelination process in the central nervous system (CNS). Complex interactions of various "environmental or infectious" factors may act as triggers in autoimmunity and disease progression. The association between viral infections, especially human immunodeficiency virus (HIV) and MS is one potential cause that is not well understood. This study aims to summarize the available data on human HIV infection in MS disease progression. In this study, the keywords "Multiple sclerosis", "Human immunodeficiency virus ", and "Central nervous system" in the databases PubMed, and Google Scholar between 2017 and 2022 were searched and 15 articles were chosen, studied, and analyzed. Revealed histologic signs of "MS-like illness" in the setting of HIV, which comprised widespread demyelination with reactive astrocytes, foamy macrophages, and perivascular infiltration with inflammatory cells, all of which are compatible with MS lesions. Human immunodeficiency virus causes dysfunction of the immune system, especially characterized by hypergammaglobulinemia and chronic activation of B cells. Activation of B cells leads to increased synthesis of immunoglobulin and finally to an excess of free light chains. Free light chains may be involved in autoimmune responses against neurons. There is a high expression of HIV during the course of MS, which indicates the relationship between HIV and MS, that this virus can play a role in the development of MS by creating an inflammatory state. Therefore, measures to modulate the expression of HIV may be effective in reducing inflammatory processes in demyelinated areas of MS patients.

Keywords: multiple sclerosis, human immunodeficiency virus, central nervous system, autoimmunity

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8481 Seroprevalence of Herpes Simplex Virus and Rubella Confection in Tropical Regions in Bihar, India

Authors: Bhawana, Roshan Kamal Topno, Maneesh Kumar, Major Madhukar, Krishna Pandey, Ganesh Chandra Sahoo, Manas Ranjan Dikhit, Surya Suman, Devendra Prasad Yadav, Rishikesh Kumar, Pradeep Das

Abstract:

Viral co-infection is now very common across taxa and environments that are involved in congenital infections. Herpes simplex virus (HSV) and Rubella are the two serious viral infections, well categorized in TORCH Syndrome. Here we had endeavoured the seroprevalence of co-infection of HSV and Rubella. Systematic tests have been performed to check the virulence pattern of the co-infection. The study was conducted at Department of Virology, Rajendra Memorial Research Institute of Medical Sciences (ICMR), Patna, Bihar, India during January 2018-July 2018. 299 newly cases were attended with the sign and symptoms of HSV and Rubella. After taking written consent forms from all the subjects, blood samples were collected for serological detection. ELISA was performed to detect the presence of IgM antibody level. 12 patients were found to be IgM positive from each HSV and Rubella infection. The findings of our study showed that 6 patients were positive for both HSV and rubella and hence were co-infected. Such co-infection causes severe health problems as it leads to the mortality rate of the patients during viral infectivity. Epidemiologically, proper screening should be needed to check any chance of occurrence of such co-infection in the affected regions in large scale and take suitable preventive approach to decrease the case totality. Concern has to be given to aid proper diagnosis and treatment in order to decrease the spread of HSV and Rubella co-infection.

Keywords: HSV, Rubella, seroprevalence, co-infection, ELISA, viral infectivity

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8480 Blindness and Deafness, the Outcomes of Varicella Zoster Virus Encephalitis in HIV Positive Patient

Authors: Hadiseh Hosamirudsari, Farhad Afsarikordehmahin, Pooria Sekhavatfar

Abstract:

Concomitant cortical blindness and deafness that follow varicella zoster virus (VZV) infection is rare. We describe a case of ophthalmic zoster that caused cortical blindness and deafness after central nervous system (CNS) involvement. A 42-year old, HIV infected woman has developed progressive blurry vision and deafness, 4 weeks after ophthalmic zoster. A physical examination and positive VZV polymerase chain reaction (PCR) of cerebrospinal fluid (CSF) suggested VZV encephalitis. Complication of VZV encephalitis is considered as the cause of blindness and deafness. In neurological deficit patient especially with a history of herpes zoster, VZV infection should be regarded as the responsible agent in inflammatory disorders of nervous system. The immunocompromised state of patient (including HIV) is as important an agent as VZV infection in developing the disease.

Keywords: blindness, deafness, hiv, VZV encephalitis

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8479 Double-Spear 1-H2-1 Oncolytic-Immunotherapy for Refractory and Relapsing High-Risk Human Neuroblastoma and Glioma

Authors: Lian Zeng

Abstract:

Double-Spear 1-H2-1 (DS1-H2-1) is an oncolytic virus and an innovative biological drug candidate. The chemical composition of the drug product is a live attenuated West Nile virus (WNV) containing the human T cell costimulator (CD86) gene. After intratumoral injection, the virus can rapidly self-replicate in the injected site and lyse/kill the tumor by repeated infection among tumor cells. We also established xenograft tumor models in mice to evaluate the drug candidate's efficacy on those tumors. The results from preclinical studies on transplanted tumors in immunodeficient mice showed that DS1-H2-1 had significant oncolytic effects on human-origin cancers: it completely (100%) shrieked human glioma; limited human neuroblastoma growth reached as high as 95% growth inhibition rate (%TGITW). The safety data of preclinical animal experiments confirmed that DS1-H2-1 is safe as a biological drug for clinical use. In the preclinical drug efficacy experiment, virus-drug administration with different doses did not show abnormal signs and disease symptoms in more than 300 tested mice, and no side effects or death occurred through various administration routes. Intravenous administration did not cause acute infectious disease or other side effects. However, the replication capacity of the virus in tumor tissue via intravenous administration is only 1% of that of direct intratumoral administration. The direct intratumoral administration of DS1-H2-1 had a higher rate of viral replication. Therefore, choosing direct intratumoral injection can ensure both efficacy and safety.

Keywords: oncolytic virus, WNV-CD86, immunotherapy drugs, glioma, neuroblastoma

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8478 Diffraction-Based Immunosensor for Dengue NS1 Virus

Authors: Harriet Jane R. Caleja, Joel I. Ballesteros, Florian R. Del Mundo

Abstract:

The dengue fever belongs to the world’s major cause of death, especially in the tropical areas. In the Philippines, the number of dengue cases during the first half of 2015 amounted to more than 50,000. In 2012, the total number of cases of dengue infection reached 132,046 of which 701 patients died. Dengue Nonstructural 1 virus (Dengue NS1 virus) is a recently discovered biomarker for the early detection of dengue virus. It is present in the serum of the dengue virus infected patients even during the earliest stages prior to the formation of dengue virus antibodies. A biosensor for the dengue detection using NS1 virus was developed for faster and accurate diagnostic tool. Biotinylated anti-dengue virus NS1 was used as the receptor for dengue virus NS1. Using the Diffractive Optics Technology (dotTM) technique, real time binding of the NS1 virus to the biotinylated anti-NS1 antibody is observed. The dot®-Avidin sensor recognizes the biotinylated anti-NS1 and this served as the capture molecule to the analyte, NS1 virus. The increase in the signal of the diffractive intensity signifies the binding of the capture and the analyte. The LOD was found to be 3.87 ng/mL while the LOQ is 12.9 ng/mL. The developed biosensor was also found to be specific for the NS1 virus.

Keywords: avidin-biotin, diffractive optics technology, immunosensor, NS1

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8477 Prevalence and Associated Factors of Chronic Energy Malnutrition among Human Immune Deficiency Virus Infected Pregnant Women in Health Centers of Addis Ababa, Ethiopia

Authors: Getachew Adugna

Abstract:

Background: Chronic energy malnutrition and human immune deficiency virus among pregnant women are highly prevalent in Sub-Saharan Africa, and they are interrelated in a vicious cycle. However, the prevalence of chronic energy malnutrition and its determinant factors among human immune deficiency virus-positive pregnant women is not well studied in Ethiopia and Addis Ababa in particular. Objective: To determine the prevalence & associated factors of chronic energy malnutrition among human immune deficiency virus-positive pregnant women in health centres of Addis Ababa Ethiopia. Methods: An institution-based cross-sectional study was conducted and a systematic random sampling technique was used to select study subjects. A total of 253 study subjects were enrolled in the study—a structured and pre-tested questionnaire collected sociodemographic, maternal health-related, and nutritional-related variables. MUAC measurements were taken and medical charts were reviewed. Bi-variable and multi-variable logistic regression analyses were used to assess the effect of different factors on chronic energy malnutrition. Result: The overall prevalence of chronic energy malnutrition was 32.0%. It was significantly associated with dietary counselling (AOR: 0.062; 95%CI: 0.007, 0.549), CD4 level (AOR: 0.219; 95%CI: 0.025, 1.908), and clinical stage (AOR: 0.127; 95%CI: 0.053, 0.305). Conclusions: The prevalence of chronic energy malnutrition among Human Immune deficiency virus-infected pregnant women in Addis Ababa was high and Nutritional Intervention should be an integral part of the HIV care program.

Keywords: chronic energy malnutrition, HIV, MUAC, Addis Ababa

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8476 Trends in the Incidence of Bloodstream Infections in Patients with Hematological Malignancies in the Period 1991–2012

Authors: V. N. Chebotkevich, E. E. Schetinkina, V. V. Burylev, E. I. Kaytandzhan, N. P. Stizhak

Abstract:

Objective: Blood stream infections (BSI) are severe, life-threatening illness for immuno compromised patients with hematological malignancies. We report the trend in blood-stream infections in this group of patients in the period 1991-2013. Methods: A total of 4742 blood samples investigated. All blood cultures were incubated in a continuous monitoring system for 7 days before discarding negative. On signaled positive, organism was identified by conventional methods. The Real-time polymerase chain reaction (PCR) was used for the indication of human herpes virus 6 (HHV-6), Cytomegalovirus (CMV) and Epstein-Barr virus (EBV). Results: Between 1991 and 2001 the incidence of Gram-positive bacteria (Staphylococcus epidermidis, Staphylococcus aureus) being the most common germs isolated (70,9%) were as Gram-negative rods (Escherichia coli, Klebsiella spp., Pseudomonas spp.) – 29,1%. In next decade 2002-2012 the number of Gram-negative bacteria was increased up to 40.2%. It is shown that the incidence of bacteremia was significantly more frequent at the background of detectable Cytomegalovirus and Epstein-Barr virus-specific DNA in blood. Over recent years, an increased frequency of micro mycetes was registered in blood of the patients with hematological malignancies (Candida spp. was predominant). Conclusion: Accurate and timely detection of BSI is important in determining appropriate treatment of infectious complications in patients with hematological malignancies. The isolation of Staphylococcus epidermidis from blood cultures remains a clinical dilemma for physicians and microbiologists. But in many cases this agent is of the clinical significance in immunocompromised patients with hematological malignancies. The role of CMV and EBV in development of bacteremia was demonstrated.

Keywords: infectious complications, blood stream infections, bacteremia, hemoblastosis

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8475 Development of Biosensor Chip for Detection of Specific Antibodies to HSV-1

Authors: Zatovska T. V., Nesterova N. V., Baranova G. V., Zagorodnya S. D.

Abstract:

In recent years, biosensor technologies based on the phenomenon of surface plasmon resonance (SPR) are becoming increasingly used in biology and medicine. Their application facilitates exploration in real time progress of binding of biomolecules and identification of agents that specifically interact with biologically active substances immobilized on the biosensor surface (biochips). Special attention is paid to the use of Biosensor analysis in determining the antibody-antigen interaction in the diagnostics of diseases caused by viruses and bacteria. According to WHO, the diseases that are caused by the herpes simplex virus (HSV), take second place (15.8%) after influenza as a cause of death from viral infections. Current diagnostics of HSV infection include PCR and ELISA assays. The latter allows determination the degree of immune response to viral infection and respective stages of its progress. In this regard, the searches for new and available diagnostic methods are very important. This work was aimed to develop Biosensor chip for detection of specific antibodies to HSV-1 in the human blood serum. The proteins of HSV1 (strain US) were used as antigens. The viral particles were accumulated in cell culture MDBK and purified by differential centrifugation in cesium chloride density gradient. Analysis of the HSV1 proteins was performed by polyacrylamide gel electrophoresis and ELISA. The protein concentration was measured using De Novix DS-11 spectrophotometer. The device for detection of antigen-antibody interactions was an optoelectronic two-channel spectrometer ‘Plasmon-6’, using the SPR phenomenon in the Krechman optical configuration. It was developed at the Lashkarev Institute of Semiconductor Physics of NASU. The used carrier was a glass plate covered with 45 nm gold film. Screening of human blood serums was performed using the test system ‘HSV-1 IgG ELISA’ (GenWay, USA). Development of Biosensor chip included optimization of conditions of viral antigen sorption and analysis steps. For immobilization of viral proteins 0.2% solution of Dextran 17, 200 (Sigma, USA) was used. Sorption of antigen took place at 4-8°C within 18-24 hours. After washing of chip, three times with citrate buffer (pH 5,0) 1% solution of BSA was applied to block the sites not occupied by viral antigen. It was found direct dependence between the amount of immobilized HSV1 antigen and SPR response. Using obtained biochips, panels of 25 positive and 10 negative for the content of antibodies to HSV-1 human sera were analyzed. The average value of SPR response was 185 a.s. for negative sera and from 312 to. 1264 a.s. for positive sera. It was shown that SPR data were agreed with ELISA results in 96% of samples proving the great potential of SPR in such researches. It was investigated the possibility of biochip regeneration and it was shown that application of 10 mM NaOH solution leads to rupture of intermolecular bonds. This allows reuse the chip several times. Thus, in this study biosensor chip for detection of specific antibodies to HSV1 was successfully developed expanding a range of diagnostic methods for this pathogen.

Keywords: biochip, herpes virus, SPR

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8474 The Prevalence of Blood-Borne Viral Infections among Autopsy Cases in Jordan

Authors: Emad Al-Abdallat, Faris G. Bakri, Azmi Mahafza, Rayyan Al Ali, Nidaa Ababneh, Ahmed Idhair

Abstract:

Background: Morgues are high-risk areas for the spread of infection from the cadavers to the staff during the postmortem examination. Infection can spread from corpses to workers by the airborne route, by direct contact, or from needle and sharp object injuries. Objective: Knowledge about the prevalence of these infections among autopsies is prudent to appreciate any risk of transmission and to further enforce safety measures. Method: A total of 242 autopsies were tested. Age ranged from 3 days to 94 years (median 75.5 years, mean 45.3 (21.9 ± SD)). There were 172 (71%) males. Results: The cause of death was considered natural in 137 (56.6%) cases, accidental in 89 (36.8%), homicidal in 9 (3.7%), suicidal in 4 (1.7%), and unknown in 3 (1.2%). Hepatitis B surface antigen was positive in 5 (2.1%) cases. Hepatitis C virus antibody was detected in 5 (2.1%) cases and the hepatitis C virus polymerase chain reaction was positive in 2 of them (0.8%). HIV antibody was not detected in any of the cases. Conclusions: Autopsies can be associated with exposure to blood borne viruses. Autopsies performed during the study period were tested for hepatitis B surface antigen, hepatitis C virus antibody, and human immunodeficiency virus antibody. Positive tests were subsequently confirmed by polymerase chain reaction. There is low prevalence of infections with these viruses in our autopsy cases. However, the risk of transmission remains a threat. Healthcare workers in the forensic departments should adhere to standard precautions.

Keywords: autopsy, hepatitis B virus, hepatitis C virus, human immunodeficiency virus, Jordan

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8473 A Comparative Study of Virus Detection Techniques

Authors: Sulaiman Al amro, Ali Alkhalifah

Abstract:

The growing number of computer viruses and the detection of zero day malware have been the concern for security researchers for a large period of time. Existing antivirus products (AVs) rely on detecting virus signatures which do not provide a full solution to the problems associated with these viruses. The use of logic formulae to model the behaviour of viruses is one of the most encouraging recent developments in virus research, which provides alternatives to classic virus detection methods. In this paper, we proposed a comparative study about different virus detection techniques. This paper provides the advantages and drawbacks of different detection techniques. Different techniques will be used in this paper to provide a discussion about what technique is more effective to detect computer viruses.

Keywords: computer viruses, virus detection, signature-based, behaviour-based, heuristic-based

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8472 Addressing Factors Associated with Vertical HIV Transmission among Pregnant Women in Rwanda

Authors: Murorunkwere Marie Claire

Abstract:

Introduction: In Sub-Saharan Africa and specifically in Rwandan rural areas, mother-to-Child human immunodeficiency virus transmission remains a big challenge. This is mainly due to lack of awareness and ignorance among pregnant rural women, leading to neglect regular taking of prophylactic antiretroviral treatment and to persistently beliefs in traditional healers and home deliveries. This paper explores the factors associated with stagnant reduction in human immunodeficiency virus vertical transmission among pregnant rural women and provides solutions to tackle it. Methodology: The first phase of this research will be a qualitative survey was conducted to assess the knowledge, attitudes and practices towards vertical human immunodeficiency virus transmission among pregnant women in one rural district in Rwanda. The data generated from phase one of this research will be used to address the main factors revealed through community mobilization and motivation on attending required antenatal consultations and hospital deliveries, proper and regular antiretroviral treatment taking, and discouraging beliefs in traditional healers and home deliveries. Refresher training seminars will also be organized for healthcare providers qualified on conducting deliveries about current measures to maximize the reduction of chances that can lead to mother -child contamination (to avoid early rupture of membranes and to prevent any source of contamination). Results: This paper is expected to contribute in a significant reduction of the vertical human immunodeficiency virus transmission burden among pregnant rural women. Conclusion: Strong campaigns on prevention of mother- to-child human immunodeficiency virus transmission and community mobilization of pregnant rural women, and house to house education and continuous reminders as well as training seminars to health care personnel on updated measures is, key in addressing vertical human immunodeficiency virus transmission.

Keywords: attitudes transformation, community mobilisation, pregnant rural women, vertical HIV transmission

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8471 The Discussion of Peritoneal Dialysis Patients Taking Proper Portion of Valacyclovir

Authors: Wan Shan Chiang, Charn Ting Wang, Wei-Chih Kan, Hui-Chen Huang

Abstract:

Dialysis patients have risk in Zoster virus because of low immune. Valacyclovir (product name: Valtex) 500mg/tab, an anti-zoster virus medicine, is digested in kidney and it has side-effect of nervous system in patients with malfunction kidneys. Although the clinical basis of the proposed administration, we found that patients still have side effects. So we want to explore the appropriate dose of peritoneal dialysis patients. We read small samples of case reports and analyze 8 cases in our hospital, some patients’ Kt/v, match the standard of dialysis, and still go to the toilet, they still have side effect seriously with 500mg portion. The solution to this includes stopping medicine, reduction of medicine, increase of liquid change and timely hemodialysis and all of them speed up the recovery. The safety of medication needs extra attention of medical care employee. If they can tell the doctor if the patient has urine or not in his or her Kt/v, the doctor can prescribe the medicine accordingly. About the limitation, due to the lack of cases and related pharmacokinetics numbers. Therefore, for peritoneal patients, we think 500mg/48hoursis the saves. We also want to remind pharmaceuticals to revise the portion taken by patients, so that the doctor may judge the use.

Keywords: herpes zoster, Valacyclovir, peritoneal dialysis, health education

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8470 Facile Synthetic Process for Lamivudine and Emtricitabine

Authors: Devender Mandala, Paul Watts

Abstract:

Cis-Nucleosides mainly lamivudine (3TC) and emtricitabine (FTC) are an important tool in the treatment of Human immune deficiency virus (HIV), Hepatitis B virus (HBV) and Human T-Lymotropoic virus (HTLV). Lamivudine and emtricitabine are potent nucleoside analog reverse transcriptase inhibitors (nRTI). These two drugs are synthesized by a four-stage process from the starting materials: menthyl glyoxylate hydrate and 1,4-dithane-2,5-diol to produce the 5-hydroxy oxathiolane which upon acetylation with acetic anhydride to yield 5-acetoxy oxathiolane. Then glycosylation of this acetyl product with silyl protected nucleoside to produce the intermediate. The reduction of this intermediates can provide the final targets. Although there are several different methods reported for the synthesis of lamivudine and emtricitabine as a single enantiomer, we required an efficient route, which was suitable for large-scale synthesis to support the development of these compounds. In this process, we successfully prepared the intermediates of lamivudine and emtricitabine without using any solvents and catalyst, thus promoting the green synthesis. All the synthesized compound were confirmed by TLC, GC, Mass, NMR and 13C NMR spectroscopy.

Keywords: emtricitabine, green synthesis, lamivudine, nucleoside

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8469 Zika Virus NS5 Protein Potential Inhibitors: An Enhanced in silico Approach in Drug Discovery

Authors: Pritika Ramharack, Mahmoud E. S. Soliman

Abstract:

The re-emerging Zika virus is an arthropod-borne virus that has been described to have explosive potential as a worldwide pandemic. The initial transmission of the virus was through a mosquito vector, however, evolving modes of transmission has allowed the spread of the disease over continents. The virus already been linked to irreversible chronic central nervous system (CNS) conditions. The concerns of the scientific and clinical community are the consequences of Zika viral mutations, thus suggesting the urgent need for viral inhibitors. There have been large strides in vaccine development against the virus but there are still no FDA-approved drugs available. Rapid rational drug design and discovery research is fundamental in the production of potent inhibitors against the virus that will not just mask the virus, but destroy it completely. In silico drug design allows for this prompt screening of potential leads, thus decreasing the consumption of precious time and resources. This study demonstrates an optimized and proven screening technique in the discovery of two potential small molecule inhibitors of Zika virus Methyltransferase and RNA-dependent RNA polymerase. This in silico “per-residue energy decomposition pharmacophore” virtual screening approach will be critical in aiding scientists in the discovery of not only effective inhibitors of Zika viral targets, but also a wide range of anti-viral agents.

Keywords: NS5 protein inhibitors, per-residue decomposition, pharmacophore model, virtual screening, Zika virus

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8468 Nutritional Status of People Living with Human Immuno Virus/Acquired Immune Deficiency Syndrome Attending Anti-Retro Viral Treatment Clinic of BP Koirala Institute of Health Sciences, Nepal

Authors: Ghimire K., Mehta R. S., Parajuli P., Chettri R.

Abstract:

Background: Malnutrition is a common hallmark of Human Immuno Virus (HIV) disease. It plays a synergistic role in immunosuppression which is initiated by Human Immuno Virus itself, and malnutrition forms an independent risk factor for disease progression. Objectives: The objective of the study is to assess the nutritional status of the people living with Human Immuno Virus/Acquired Immune Deficiency Syndrome attending the Anti-Retro viral Treatment Clinic and find the association of nutritional status with different socio-demographic variables. Methods: A total of 101 people living with HIV/AIDS (PLWHA) were selected by convenient sampling technique. The study was conducted at the ART clinic of BPKIHS. A subjective global assessment tool was used for data collection. Descriptive and inferential statistics were used for data analysis. Results: The study demonstrated that the mean age of the respondents was 40.97+8.650 years. 65.3% were well-nourished, and 34.7% of the participants were mildly/moderately malnourished, whereas none of them were severely malnourished. BMI was statistically significant with education status, family income, and duration of illness of the participants, and nutritional status was statistically significant with gender, marital status, education status, and family history of HIV. Conclusion: On the basis of the result, it can be concluded that more than half of the respondents were well nourished. Gender, marital status, and education are associated with nutritional status.

Keywords: nutritional status, people living with HIV/AIDS, ART treatment, Nepal

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8467 Biopsy Proven Polyoma (BK) Virus in Saudi Kidney Recipients – Prevalence, Clinicopathological Features and Clinico-Pathological Correlations

Authors: Sarah Hamdan Al-Jahdali, Khaled Alsaad, Abdullah Al-Sayyari

Abstract:

Objectives: To study the prevalence, clinicopathological features, risk factors and outcome of biopsy proven polyoma (BK) virus infection among Saudi kidney transplant recipients and compare them to negative BK virus group. Methods: We retrospectively reviewed the charts of all the patients with biopsy-proven polyoma (BK) virus infection in King Abdulaziz Medical City in Riyadh between 2005 and 2011. The details of clinical presentation, the indication for kidney biopsy, the laboratory findings at presentation, the natural history of the disease, thepathological findings, the prognosis as well as the response to therapy were all recorded. Results: Kidney biopsy was performed in 37 cases of unexplained graft dysfunction. BK virus was found in 10 (27%). Out of those 10, 3 (30%) ended with graft failure. BK virus occurred in all patients who received ATG induction therapy 100% versus 59.3% in the non BK virus patients (p=0.06). Furthermore, the risk of BK virus was much less in those who received acyclovir as an anti-viral prophylaxis as compared to those who did not receive it (p=0.01). Also, patients with BK virus weighed much less (mean 46.7±20.6 Kgs) than those without BK virus at time of transplantation (mean 64.3±12.1). Graft survival was better among deceased donor kidneys compared to living ones (P=0.016) and with older age (P=0.005). Conclusion: Our findings suggest the involvement of ATG induction therapy, the lack of antiviral prophylaxis therapy and lower weight at transplant as significant risk factors for the development of BK virus infection.

Keywords: BKVAN, BKV, kidney transpant, Saudi Arabia

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8466 Household Low Temperature MS2 (ATCC15597-B1) Virus Inactivation Using a Hot Bubble Column Evaporator

Authors: Adrian Garrido Sanchis, Richard Pashley

Abstract:

The MS2 (ATCC15597-B1) virus was used as a surrogate to estimate the inactivation rates for enteric viruses when using a hot air bubble column evaporator (HBCE) system in the treatment of household wastewater. In this study, we have combined MS2 virus surface charging properties with thermal inactivation rates, using an improved double layer plaque assay technique, in order to assess the efficiency of the HBCE process for virus removal in water. When bubbling a continuous flow of dry air, at 200°C, only heats the aqueous solution in the bubble column to about 50°C. Viruses are not inactivated by this solution temperature, as confirmed separately from water bath heating experiments. Hence, the efficiency of the HBCE process for virus removal in water appeared to be caused entirely by collisions between the hot air bubbles and the virus organisms. This new energy efficient treatment for water reuse applications can reduce the thermal energy required to only 25% (about 113.7 kJ/L) of that required for boiling (about 450 kJ/L).

Keywords: MS2 virus inactivation, water reuse, hot bubble column evaporator, water treatment

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8465 Prevalence of High Risk Human Papillomavirus in Cervical Dysplasia and Cancer Samples from Twin Cities in Pakistan

Authors: Sana Gul, Sheeba Murad, Aneela Javed

Abstract:

Introduction: Human Papilloma Virus (HPV) is small DNA virus mostly infecting mucosa and cutaneous keratinocytes. So far, more than 200 Human papillomaviruses are known. HPV have been divided into high- and low-risk on the basis of their oncogenic potential. High risk HPV is considered to be the main etiological cause for cervical cancer. Objective: Current study was designed to screen the local cervical cancer patients from the twin cities of Pakistan for the occurance of high risk HPV. Methodology: A total of 67 formalin fixed paraffin-embedded samples of cervical cancer biopsies were obtained from the government hospitals in Islamabad and Rawalpindi. Cervical cancer biopsies were examined for the presence of HPV DNA. Polymerase chain reaction (PCR) was used for the amplification of a region in the HPV-L1 gene for the general detection of the Papilloma virus and for the genotype specific detection of high risk HPV 16 and 18 using the GP5/GP6 primers and genotype specific primers respectively. Results: HPV DNA was detected in 59 out of 67 samples analyzed. 30 samples showed the presence of HPV16 while 22 samples were positive for HPV 18 . HPV subtype could not be determined in 7 samples. Conclusion: Our results show a strong association between HPV infection and cervical cancer among women in twin cities of Pakistan. One way to minimize the disease burden in relation to HPV infection in Pakistani population is the use of prophylactic vaccines and routine screening. An early diagnosis of HPV infection will allow better health management to reduce the risk of developing cervical cancer.

Keywords: cervical cancer, Pakistan, human papillomavirus, HPV 16

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8464 Survey of Potato Viral Infection Using Das-Elisa Method in Georgia

Authors: Maia Kukhaleishvili, Ekaterine Bulauri, Iveta Megrelishvili, Tamar Shamatava, Tamar Chipashvili

Abstract:

Plant viruses can cause loss of yield and quality in a lot of important crops. Symptoms of pathogens are variable depending on the cultivars and virus strain. Selection of resistant potato varieties would reduce the risk of virus transmission and significant economic impact. Other way to avoid reduced harvest yields is regular potato seed production sampling and testing for viral infection. The aim of this study was to determine the occurrence and distribution of viral diseases according potato cultivars for further selection of virus-free material in Georgia. During the summer 2015- 2016, 5 potato cultivars (Sante, Laura, Jelly, Red Sonia, Anushka) at 5 different farms located in Akhalkalaki were tested for 6 different potato viruses: Potato virus A (PVA), Potato virus M (PVM), Potato virus S (PVS), Potato virus X (PVX), Potato virus Y (PVY) and potato leaf roll virus (PLRV). A serological method, Double Antibody Sandwich-Enzyme linked Immunosorbent Assay (DASELISA) was used at the laboratory to analyze the results. The result showed that PVY (21.4%) and PLRV (19.7%) virus presence in collected samples was relatively high compared to others. Researched potato cultivars except Jelly and Laura were infected by PVY with different concentrations. PLRV was found only in three potato cultivars (Sante, Jelly, Red Sonia) and PVM virus (3.12%) was characterized with low prevalence. PVX, PVA and PVS virus infection was not reported. It would be noted that 7.9% of samples were containing PVY/PLRV mix infection. Based on the results it can be concluded that PVY and PLRV infections are dominant in all research cultivars. Therefore significant yield losses are expected. Systematic, long-term control of potato viral infection, especially seed-potatoes, must be regarded as the most important factor to increase seed productivity.

Keywords: virus, potato, infection, diseases

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8463 Eradication of Apple mosaic virus from Corylus avellana L. via Cryotherapy and Confirmation of Virus-Free Plants via Reverse Transcriptase Polymerase Chain Reaction

Authors: Ergun Kaya

Abstract:

Apple mosaic virus (ApMV) is an ilarvirus causing harmful damages and product loses in many plant species. Because of xylem and phloem vessels absence, plant meristem tissues used for meristem cultures are virus-free, but sometimes only meristem cultures are not sufficient for virus elimination. Cryotherapy, a new method based on cryogenic techniques, is used for virus elimination. In this technique, 0.1-0.3mm meristems are excised from organized shoot apex of a selected in vitro donor plant and these meristems are frozen in liquid nitrogen (-196 °C) using suitable cryogenic technique. The aim of this work was to develop an efficient procedure for ApMV-free hazelnut via cryotherapy technique and confirmation of virus-free plants using Reverse Transcriptase-PCR technique. 100% virus free plantlets were obtained using droplet-vitrification method involved cold hardening in vitro cultures of hazelnut, 24 hours sucrose preculture of meristems on MS medium supplemented with 0.4M sucrose, and a 90 min PVS2 treatment in droplets.

Keywords: droplet vitrification, hazelnut, liquid nitrogen, PVS2

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8462 Oncolytic Efficacy of Thymidine Kinase-Deleted Vaccinia Virus Strain Tiantan (oncoVV-TT) in Glioma

Authors: Seyedeh Nasim Mirbahari, Taha Azad, Mehdi Totonchi

Abstract:

Oncolytic viruses, which only replicate in tumor cells, are being extensively studied for their use in cancer therapy. A particular virus known as the vaccinia virus, a member of the poxvirus family, has demonstrated oncolytic abilities glioma. Treating Glioma with traditional methods such as chemotherapy and radiotherapy is quite challenging. Even though oncolytic viruses have shown immense potential in cancer treatment, their effectiveness in glioblastoma treatment is still low. Therefore, there is a need to improve and optimize immunotherapies for better results. In this study, we have designed oncoVV-TT, which can more effectively target tumor cells while minimizing replication in normal cells by replacing the thymidine kinase gene with a luc-p2a-GFP gene expression cassette. Human glioblastoma cell line U251 MG, rat glioblastoma cell line C6, and non-tumor cell line HFF were plated at 105 cells in a 12-well plates in 2 mL of DMEM-F2 medium with 10% FBS added to each well. Then incubated at 37°C. After 16 hours, the cells were treated with oncoVV-TT at an MOI of 0.01, 0.1 and left in the incubator for a further 24, 48, 72 and 96 hours. Viral replication assay, fluorescence imaging and viability tests, including trypan blue and crystal violet, were conducted to evaluate the cytotoxic effect of oncoVV-TT. The finding shows that oncoVV-TT had significantly higher cytotoxic activity and proliferation rates in tumor cells in a dose and time-dependent manner, with the strongest effect observed in U251 MG. To conclude, oncoVV-TT has the potential to be a promising oncolytic virus for cancer treatment, with a more cytotoxic effect in human glioblastoma cells versus rat glioma cells. To assess the effectiveness of vaccinia virus-mediated viral therapy, we have tested U251mg and C6 tumor cell lines taken from human and rat gliomas, respectively. The study evaluated oncoVV-TT's ability to replicate and lyse cells and analyzed the survival rates of the tested cell lines when treated with different doses of oncoVV-TT. Additionally, we compared the sensitivity of human and mouse glioma cell lines to the oncolytic vaccinia virus. All experiments regarding viruses were conducted under biosafety level 2. We engineered a Vaccinia-based oncolytic virus called oncoVV-TT to replicate specifically in tumor cells. To propagate the oncoVV-TT virus, HeLa cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 10 MOI virus was added. After 48 h, cells were harvested by scraping, and viruses were collected by 3 sequential freezing and thawing cycles followed by removal of cell debris by centrifugation (1500 rpm, 5 min). The supernatant was stored at −80 ◦C for the following experiments. To measure the replication of the virus in Hela, cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 5 MOI virus or equal dilution of PBS was added. At the treatment time of 0 h, 24 h, 48 h, 72 h and 96 h, the viral titers were determined under the fluorescence microscope (BZ-X700; Keyence, Osaka, Japan). Fluorescence intensity was quantified using the imagej software according to the manufacturer’s protocol. For the isolation of single-virus clones, HeLa cells seeded in six-well plates (5×105 cells/well). After 24 h (100% confluent), the cells were infected with a 10-fold dilution series of TianTan green fluorescent protein (GFP)virus and incubated for 4 h. To examine the cytotoxic effect of oncoVV-TT virus ofn U251mg and C6 cell, trypan blue and crystal violet assay was used.

Keywords: oncolytic virus, immune therapy, glioma, vaccinia virus

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