Development of an analytical platform for delivery of recombinant oncolytic viruses

Over the past two decades, a number of modified DNA and RNA viruses have shown great promise in cancer therapy. Their tumor specific cytotoxic activity involves several mechanisms. These include preferential uptake and replication of the virus in cancer cells, followed by infection and lysis of these cells (oncolysis). In addition, oncolytic viruses (OV), especially those engineered with appropriate transgenes, have been shown to induce tumor specific immune responses, thereby opening up an exciting area of cancer immunotherapy. One OV, containing a recombinant Herpes Simplex Virus (HSV) with a Granulocyte Macrophage Colony Stimulating Factor (GMCSF) transgene insert, was approved in 2015 for treatment of refractory melanoma. While this drug, known as T-Vec or Imlygic, is administered intratumorally, several other OVs are in pre-clinical and clinical development with intravenous (IV) administration as an objective. The IV route potentially offers the advantages of accessing metastatic sites and eliciting systemic immunotherapeutic response. A recombinant form of the Newcastle Disease Virus (rNDV) has been recently developed that also incorporates GMCSF as a transgene. NDV is a single stranded, negative sense, enveloped RNA virus and, in native form, is not a human but an avian pathogen. Genetic engineering was performed to reduce avian pathogenicity and increase immunotherapeutic potential against cancer [1]. In addition to lysis of cancer cells produced by viral infection, rNDV has demonstrated induction of cytokine and chemokine responses, cell infiltration and long - term suppression of tumor growth in animal models. Production of recombinant viruses requires selection of appropriate host cells, based on considerations of yield and quality of the product. In recent years, regulatory agencies have been open to use of human cell lines as host, including some tumor derived cells, such as HeLa. An advantage offered by HeLa cells is potential incorporation into the virus of host proteins known as regulators of complement activation (RCA). This allows a longer half-life of the virus in circulation. However, impurities derived from host cells as well as process conditions must be kept to the lowest levels possible. A thorough analytical and biophysical evaluation of the purified virus particles includes measurements of (a) infectivity, (b) morphology, (c) size distribution, (c) genetic integrity, and (d) major viral proteins. In addition, levels of contaminants such as residual host cell proteins and DNA must be evaluated, including sizes of residual DNA fragments. Robust and accurate measurement of infectious titer is especially important as this determines doses of an OV to be delivered to patients. It also provides a measure of virus stability and is critical to supporting process development. We have optimized a Fluorescent Focus Assay (FFA) that produces concordant results with Plaque Assay (PA) over several orders of magnitude of infectious titers [2]. FFA provides a faster turn-around in a higher throughput format compared to PA. These and additional structure-function assays were developed to support process development and deliver a well-characterized OV for cancer immunotherapy. Acknowledgment: Contributions of Benjamin S Rush, Melissa L Coughlin and Jenny Sun of MedImmune to analytical development of rNDV are gratefully acknowledged. References: [1]. Cheng, X., Wang, W., Xu, Q., Harper, J., Carroll, D., Galinski, M.S., Jin, H*. 2016. Generation of a potent nonvirulent Newcastle disease virus for cancer therapy. J. Virol. 90, 5343–5352. [2]. Rush, B.S., Coughlin, M.L., Sanyal, G*. 2018. In vitro infectivity of oncolytic Newcastle Disease Virus: Correlation between plaque and fluorescent focus assays. J. Virol. Methods 251, 69–74. 1Includes work performed at Biopharmaceutical Development, MedImmune LLC, Gaithersburg, MD, US

Biometric Steganography using Variable Length Embedding

Recent growth in digital multimedia technologies has presented a lot of facilities in information transmission, reproduction and manipulation. So the concept of information security is one of the superior articles in the present day situation. The biometric information security is one of the information security mechanisms. It has the advantages and disadvantages also. The biometric system is at risk to a range of attacks. These attacks are anticipated to bypass the security system or to suspend the normal functioning. Various hazards have been discovered while using biometric system. Proper use of steganography greatly reduces the risks in biometric systems from the hackers. Steganography is one of the fashionable information hiding technique. The goal of steganography is to hide information inside a cover medium like text, image, audio, video etc. through which it is not possible to detect the existence of the secret information. Here in this paper a new security concept has been established by making the system more secure with the help of steganography along with biometric security. Here the biometric information has been embedded to a skin tone portion of an image with the help of proposed steganographic technique

Biometric Steganography using Variable Length Embedding

Recent growth in digital multimedia technologies has presented a lot of facilities in information transmission, reproduction and manipulation. So the concept of information security is one of the superior articles in the present day situation. The biometric information security is one of the information security mechanisms. It has the advantages and disadvantages also. The biometric system is at risk to a range of attacks. These attacks are anticipated to bypass the security system or to suspend the normal functioning. Various hazards have been discovered while using biometric system. Proper use of steganography greatly reduces the risks in biometric systems from the hackers. Steganography is one of the fashionable information hiding technique. The goal of steganography is to hide information inside a cover medium like text, image, audio, video etc. through which it is not possible to detect the existence of the secret information. Here in this paper a new security concept has been established by making the system more secure with the help of steganography along with biometric security. Here the biometric information has been embedded to a skin tone portion of an image with the help of proposed steganographic technique

Biometric Steganography using Variable Length Embedding

Recent growth in digital multimedia technologies has presented a lot of facilities in information transmission, reproduction and manipulation. So the concept of information security is one of the superior articles in the present day situation. The biometric information security is one of the information security mechanisms. It has the advantages and disadvantages also. The biometric system is at risk to a range of attacks. These attacks are anticipated to bypass the security system or to suspend the normal functioning. Various hazards have been discovered while using biometric system. Proper use of steganography greatly reduces the risks in biometric systems from the hackers. Steganography is one of the fashionable information hiding technique. The goal of steganography is to hide information inside a cover medium like text, image, audio, video etc. through which it is not possible to detect the existence of the secret information. Here in this paper a new security concept has been established by making the system more secure with the help of steganography along with biometric security. Here the biometric information has been embedded to a skin tone portion of an image with the help of proposed steganographic technique

Community structure of Coleoptera in Bethuadahari Wildlife Sanctuary, West Bengal, India

We focused on the coleopteran species assemblage in a tropical deciduous forest in the Bethuadahari Wildlife Sanctuary, West Bengal, India. During a 2-year survey, we collected 56 species belonging to 13 families of Coleoptera, in varying relative abundance. Among the species, 15 belong to the family Chrysomelidae, nine to the Staphyllinidae, and four to the Coccinellidae. Our results substantiate the importance of the Bethuadahari Wildlife Sanctuary for the conservation of coleopteran insects

Ordered kinetochore assembly in the human-pathogenic basidiomycetous yeast Cryptococcus neoformans

Kinetochores facilitate interaction between chromosomes and the spindle apparatus. The formation of a metazoan trilayered kinetochore is an ordered event in which inner, middle, and outer layers assemble during disassembly of the nuclear envelope during mitosis. The existence of a similar strong correlation between kinetochore assembly and nuclear envelope breakdown in unicellular eukaryotes is unclear. Studies in the hemiascomycetous budding yeasts Saccharomyces cerevisiae and Candida albicans suggest that an ordered kinetochore assembly may not be evolutionarily conserved. Here, we utilized high-resolution time-lapse microscopy to analyze the localization patterns of a series of putative kinetochore proteins in the basidiomycetous budding yeast Cryptococcus neoformans, a human pathogen. Strikingly, similar to most metazoa but atypical of yeasts, the centromeres are not clustered but positioned adjacent to the nuclear envelope in premitotic C. neoformans cells. The centromeres gradually coalesce to a single cluster as cells progress toward mitosis. The mitotic clustering of centromeres seems to be dependent on the integrity of the mitotic spindle. To study the dynamics of the nuclear envelope, we followed the localization of two marker proteins, Ndc1 and Nup107. Fluorescence microscopy of the nuclear envelope and components of the kinetochore, along with ultrastructure analysis by transmission electron microscopy, reveal that in C. neoformans, the kinetochore assembles in an ordered manner prior to mitosis in concert with a partial opening of the nuclear envelope. Taken together, the results of this study demonstrate that kinetochore dynamics in C. neoformans is reminiscent of that of metazoans and shed new light on the evolution of mitosis in eukaryotes

Effects of antiplatelet therapy on stroke risk by brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases: subgroup analyses of the RESTART randomised, open-label trial

Background Findings from the RESTART trial suggest that starting antiplatelet therapy might reduce the risk of recurrent symptomatic intracerebral haemorrhage compared with avoiding antiplatelet therapy. Brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases (such as cerebral microbleeds) are associated with greater risks of recurrent intracerebral haemorrhage. We did subgroup analyses of the RESTART trial to explore whether these brain imaging features modify the effects of antiplatelet therapy

Comparison of the Morphology Development of Polymer-Fullerene and Polymer-Polymer Solar Cells during Solution-Shearing Blade Coating

In this work, the detailed morphology studies of polymer poly(3‐hexylthiophene‐2,5‐diyl) (P3HT):fullerene(PCBM) and polymer(P3HT):polymer naphthalene diimide thiophene (PNDIT) solar cell are presented to understand the challenge for getting high performance all‐polymer solar cells. The in situ X‐ray scattering and optical interferometry and ex situ hard and soft X‐ray scattering and imaging techniques are used to characterize the bulk heterojunction (BHJ) ink during drying and in dried state. The crystallization of P3HT polymers in P3HT:PCBM bulk heterojunction shows very different behavior compared to that of P3HT:PNDIT BHJ due to different mobilities of P3HT in the donor:acceptor glass. Supplemented by the ex situ grazing incidence X‐ray diffraction and soft X‐ray scattering, PNDIT has a lower tendency to form a mixed phase with P3HT than PCBM, which may be the key to inhibit the donor polymer crystallization process, thus creating preferred small phase separation between the donor and acceptor polymer