Application of Oncolytic Poxviruses: An Emerging Paradigm in Cancer Therapy.

Despite the significant advancement of new tools and technology in the field of medical biology and molecular biology, the challenges in the treatment of most cancer types remain constant with the problem of developing resistance toward drugs and no substantial enhancement in the overall survival rate of cancer patients. Immunotherapy has shown the most promising results in different clinical and preclinical trials in the treatment of various cancer due to its higher efficacy and minimum collateral damage in many cancer patients as compared to conventional chemotherapy and radiotherapy. An oncolytic virus is a new class of immunotherapy that can selectively replicate in tumor cells and destroy them by the process of cell lysis while exerting minimum or no effect on a normal cell. Besides this, it can also activate the host's innate immune system, which generates an anti-tumor immune response to eliminate the tumor cells. Several wild types and genetically modified viruses have been investigated to show oncolytic behavior. Vaccinia virus has been studied extensively and tested for its promising oncolytic nature on various model systems and clinical trials. Recently, several engineered vaccinia viruses have been developed that express the desired genes encoded for selective penetration in tumor cells and enhanced activation of the immune system for generating anti-tumor immunity. However, further investigation is required to prove their potential and enhance their therapeutic efficacy.

Poxviruses as Agents of Biological Warfare: The Importance of Ensuring Ethical Standards for Research with Viruses.

Historically, biological agents have been used to target various populations. One of the earliest examples could be the catastrophic effect of smallpox in Australia in the eighteenth century (as alleged by some historians). Modern biological techniques can be used to both create or provide protection against various agents of biological warfare. Any microorganism (viruses, bacteria, and fungi) or its toxins can be used as biological agents. Minnesota Department of Health has listed Smallpox (variola major) as a category A bioterrorism agent, even though it has been eradicated in 1980 through an extensive vaccination campaign. Category A agents are considered the highest risk to public health. Laboratory-associated outbreaks of poxviruses could cause unprecedented occupational hazards. Only two WHO-approved BSL-4 facilities in the United States and Russia are allowed to perform research on the variola virus. So, poxviruses present themselves as a classical case of a dual-use dilemma, since research with them can be used for both beneficial and harmful purposes. Although the importance of ethics in scientific research requires no further elaboration, ethical norms assume greater significance during experimentation with poxviruses. In this chapter, we will update the readers on the sensitive nature of conducting research with poxviruses, and how these viruses can be a source of potential biological weapons. Finally, specified ethical guidelines are explored to ensure safe research practices in virology.

The long-term impact of coronavirus disease 2019 on environmental health: a review study of the bi-directional effect.

Background: When health systems worldwide grapple with the coronavirus disease 2019 (COVID-19) pandemic, its effect on the global environment is also a significant consideration factor. It is a two-way process where the pre-COVID climate factors influenced the landscape in which the disease proliferates globally and the consequences of the pandemic on our surroundings. The environmental health disparities will also have a long-lasting effect on public health response. Main body: The ongoing research on the novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and COVID-19 must also include the role of environmental factors in the process of infection and the differential severity of the disease. Studies have shown that the virus has created positive and negative ramifications on the world environment, especially in countries most critically affected by the pandemic. Contingency measures to slow down the virus, such as self-distancing and lockdowns have shown improvements in air, water, and noise quality with a concomitant decrease in greenhouse gas emissions. On the other hand, biohazard waste management is a cause for concern that can result in negative effects on planetary health. At the peak of the infection, most attention has been diverted to the medical aspects of the pandemic. Gradually, policymakers must shift their focus to social and economic avenues, environmental development, and sustainability. Conclusion: The COVID-19 pandemic has profoundly impacted the environment, both directly and indirectly. On the one hand, the sudden halt in economic and industrial activities led to a decrease in air and water pollution, as well as a reduction in greenhouse gas emissions. On the other hand, the increased use of single-use plastics and a surge in e-commerce activities have had negative effects on the environment. As we move forward, we must consider the pandemic's long-term impacts on the environment and work toward a more sustainable future that balances economic growth and environmental protection. The study shall update the readers on the various facets of the interaction between this pandemic and environmental health with model development for long-term sustainability.

Sorptive and microbial riddance of micro-pollutant ibuprofen from contaminated water: A state of the art review.

Continuous discharge of ibuprofen, a pharmaceutical compound in local water systems is becoming a budding concern as seen from data procured from the past few decades. Increased concentrations of the compound in water reservoirs resulted in adverse effects on the environment. In order to prevent the deleterious impacts of increasing ibuprofen concentration in water bodies, application of cost effective and energy efficient elimination of ibuprofen (IBP) is needed. As a result, various techniques over time have been tested for IBP expulsion from aqueous media. However, adsorption and bioremediation are still the most realistic approaches to remove ibuprofen than conventional methods, like precipitation, reverse osmosis, ion exchange, nano-filtration etc., because of their lower initial cost, reduced electricity consumption, minimized sludge generation, local availability of precursor material etc. Various researchers have reported the applicability of the adsorption and bioremediation process in remediation of ibuprofen from water. Therefore, the present review article confers both the biosorption and bioremediation process towards IBP removal from water bodies and explicates the performances of various adsorbents and microorganisms derived from various sources. The presented review also substantially emphasizes on the effect of different parameters on sorptive uptake of ibuprofen, various isotherms and kinetic models, sorption mechanism and assessment of costs, which could enable future researchers to determine widespread use of reported adsorbents and microbes towards effective elimination of IBP from aqueous media.

N-protein-RNA interaction is a drug target in a negative strand RNA virus.

The negative strand RNA virus family contains many human pathogens. Finding new antiviral drug targets against this class of human pathogens is one of the significant healthcare needs. Nucleocapsid proteins of negative strand RNA viruses wrap the viral genomic RNA and play essential roles in gene transcription and genome replication. Chandipura virus, a member of the Rhabdoviridae family, has a negative strand RNA genome. In addition to wrapping the genomic RNA, its nucleocapsid protein interacts with the positive strand leader RNA and plays a vital role in the virus life-cycle. We have designed a peptide, based on prior knowledge and demonstrated that the peptide is capable of binding specifically to the positive strand leader RNA. When the peptide was transported inside the cell, it inhibited viral growth with IC50 values in the low micromolar range. Given the widespread occurrence of leader RNAs in negative strand RNA viruses and its interaction with the nucleocapsid protein, it is likely that this interaction could be a valid drug target for other negative strand RNA viruses.

Density of CD3+ and CD8+ cells in gingivo-buccal oral squamous cell carcinoma is associated with lymph node metastases and survival.

The tumor immune microenvironment is emerging as a critical player in predicting cancer prognosis and response to therapies. However, the prognostic value of tumor-infiltrating immune cells in Gingivo-Buccal Oral Squamous Cell Carcinoma (GBOSCC) and their association with tumor size or lymph node metastases status require further elucidation. To study the relationship of tumor-infiltrating immune cells with tumor size (T stage) and lymph node metastases (N stages), we analyzed the density of tumor-infiltrating immune cells in archived, whole tumor resections from 94 patients. We characterized these sections by immune-histochemistry using 12 markers and enumerated tumor-infiltrating immune cells at the invasive margins (IM) and centers of tumors (CT). We observed that a higher density of CD3+ cells in the IM and CT was associated with smaller tumor size (T1-T2 stage). Fewer CD3+ cells was associated with larger tumor size (T3-T4 stage). High infiltration of CD3+and CD8+ cells in IM and CT as well as high CD4+ cell infiltrates in the IM was significantly associated with the absence of lymph node metastases. High infiltrates of CD3+ and CD8+ cells in CT was associated with significantly improved survival. Our results illustrate that the densities and spatial distribution of CD3+ and CD8+ cell infiltrates in primary GBOSCC tumors is predictive of disease progression and survival. Based on our findings, we recommend incorporating immune cell quantification in the TNM classification and routine histopathology reporting of GBOSCC. Immune cell quantification in CT and IM may help predict the efficacy of future therapies.

Ibuprofen sorptive efficacy of zirconium caged date seed derived steam activated alginate beads in a static bed column.

The sorption capability of zirconium coupled sodium alginate beads of steam activated biochar derived from date seed (Zr(DSPB)Al) was explored towards Ibuprofen (IBP) removal from simulated water solution in a static bed column. The impact of governing variables viz. column bed height (5-25 cm), influent (IBP) concentration (10-30 mg L-1) and inflow rate (2-6 mL min-1) was investigated in the present study. The column experimentation reflected that with an increase in column bed length, the breakthrough curve height was increased. The maximum sorbent uptake was found to be 23.33 mg g-1 from an optimal column bed height of 20 cm, influent (IBP) concentration of 30 mg L-1 and inflow rate of 2 mL min-1 with the achievement of 94.86% of IBP removal. The bed depth service model (BDST) was studied to examine the sorbent's efficacy and it was observed that column bed height was one of the effective factors towards effective IBP sorption. The Yoon-Nelson model and Thomas model corroborated extremely well with the experimental findings. The desorption study presented a sorbent efficiency up to 5 cycles for IBP exclusion with 37.59% regeneration of the column. The investigation indicated that the novel sorbent Zr(DSPB)Al with proficient performance could be successfully applied for IBP elimination from aqueous solutions.

Immune and genomic signatures in oral (head and neck) cancer.

Head and neck squamous cell carcinoma (HNSCC) is responsible for a large number of deaths each year. Oral cancer is the most frequent subtype of HNSCC. Historically, oral cancer has been associated with an increase in the consumption of tobacco and alcohol products, seen especially in the Asian subcontinent. It has also been associated with infection by the human papilloma virus (HPV), particularly strain HPV16. Treatment usually involves a multidisciplinary approach of surgery combined with chemotherapy and radiation. The advent of immunotherapy has broadened the scope for treatment. A better immune response to the tumour can also elicit the action of other therapeutic approaches. A heightened immune response, on the other hand, can lead to resistant tumour formation through the process of immunoediting. Molecular profiling of the tumour microenvironment (TME) can provide us with better insight into the mechanism and progression of the disease, ultimately opening up new therapeutic options. High-throughput molecular profiling techniques over the past decade have enabled us to appreciate the heterogeneity of the TME. In this review, we will be describing the clinicopathological role of the immune and genomic landscape in oral cancer. This study will update readers on the several immunological and genetic factors that can play an important function as predictive and prognostic biomarkers in various forms of head and neck cancer, with a special emphasis on oral carcinoma.

Enhanced basepair dynamics pre-disposes protein-assisted flips of key bases in DNA strand separation during transcription initiation.

Localized separation of strands of duplex DNA is a necessary step in many DNA-dependent processes, including transcription and replication. Little is known about how these strand separations occur. The strand-separated E.coli RNA polymerase-promoter open-complex structure showed four bases of the non-template strand, the master base -11A, -7, -6 and +2, in a flipped state and inserted into protein pockets. To explore whether any property of these bases in the duplex state pre-disposes them to flipping, NMR studies were performed on a wild-type promoter in the duplex state. Measurement of relaxation times indicates that a limited number of base pairs, including the flipped ones, have faster opening rates than the rest. Molecular dynamics studies also show an inherently high dynamic character of the -11A:T base pair in the wild-type strand-paired state. In order to explore the role of the RNA polymerase in the flipping process, we have used 2-aminopurine as a fluorescent probe. Slower kinetics of the increase of 2-aminopurine fluorescence was observed with RNA polymerases containing several mutant σ70s. This may be interpreted as the protein playing an important role in enhancing the flipping rate. These results suggest that flipping of -11A, and perhaps other flipped bases observed in the open-complex, is facilitated by its inherent proclivity to open-up with further assistance from the protein, thus leading to a strand-open state. Other DNA-based processes that require strand-separation may use similar pathways for strand separation. We conclude that not only basepair stability, but also dynamics may play an important role in the strand-separation.

Construction & establishment of two minigenome rescue systems for Chandipura virus driven by recombinant vaccinia virus expressing T7 polymerase.

BACKGROUND & OBJECTIVES: Chandipura virus (CHPV) is an emerging pathogenic rhabdovirus with a high case fatality rate. There are no reports of a minigenome system for CHPV, which could help its study without having to use the infectious agent. This study was, therefore, undertaken for the establishment of T7 polymerase-driven minigenome system for CHPV. METHODS: The minigenome rescue system for CHPV consists of three helper plasmids expressing the nucleocapsid protein (N), phosphoprotein (P) and large protein (L) based on a recombinant vaccinia virus expressing bacteriophage T7 polymerase (vTF7-3). The minigenome construct is composed of a reporter gene, flanked by the non-coding regions of CHPV. Two minigenomes were constructed in an antigenome or complimentary sense, expressing luciferase or green fluorescent protein (GFP). The minigenome system was evaluated by co-transfection of the minigenome construct and three helper plasmids into CV-1 cells and analysis of the reporter gene activity. RESULTS: All the helper proteins were expressed from the helper plasmids confirmed by Western blotting. Expression of reporter genes was observed from both the GFP and luciferase-based minigenomes. Green fluorescence could be visualized directly in live CV-1 cells. Luciferase activity was found to be significantly different from control. INTERPRETATION & CONCLUSIONS: The results showed that the helper plasmids provided all the necessary viral structural proteins required for the production of minigenome mRNA template, which in turn could rescue the expression of reporter genes. Thus, these minigenomes can be applied to mimic the manifestation of CHPV life cycle.

A peptide targeted against phosphoprotein and leader RNA interaction inhibits growth of Chandipura virus -- an emerging rhabdovirus.

The fatal illness caused by Chandipura virus (CHPV), an emerging pathogen, presently lacks any therapeutic option. Previous research suggested that interaction between the virally encoded phosphoprotein (P) and the positive sense leader RNA (le-RNA) may play an important role in the viral lifecycle. In this report, we have identified a ß-sheet/loop motif in the C-terminal domain of the CHPV P protein as essential for this interaction. A synthetic peptide encompassing this motif and spanning a continuous stretch of 36 amino acids (Pep208-243) was found to bind the le-RNA in vitro and inhibit CHPV growth in infected cells. Furthermore, a stretch of three amino acid residues at position 217-219 was identified as essential for this interaction, both in vitro and in infected cells. siRNA knockdown-rescue experiments demonstrated that these three amino acid residues are crucial for the leader RNA binding function of P protein in the CHPV life cycle. Mutations of these three amino acid residues render the peptide completely ineffective against CHPV. Effect of inhibition of phosphoprotein-leader RNA interaction on viral replication was assayed. Peptide Pep208-243 tagged with a cell penetrating peptide was found to inhibit CHPV replication as ascertained by real time RT-PCR. The specific inhibition of viral growth observed using this peptide suggests a new possibility for designing of anti-viral agents against Mononegavirale group of human viruses.