Therapeutic targeting of Sp1 transcription factor and survivin, are studied in various cancers due to their consistent overexpression. These markers result in poorer cancer prognoses and their downregulation has been investigated as an effective treatment approach. Mithramycin-A and Tolfenamic acid are two drugs with innate anti-cancer properties and are suggested to be able to target Sp1 through GC/GT DNA binding interference, however in-depth binding and mechanistic studies are lacking. Through docking analysis, we investigated Mithramycin-A and Tolfenamic acid in terms of their specific binding interactions with Sp1 and survivin. Through further molecular dynamics simulations including Root Mean Square (RMS) Fluctuation and RMS Deviation, rGYr, and H-bond analysis, we identified critical residues involved in drug interactions with each protein in question. We show Mithramycin-A as the superior binding candidate to each protein and found that it exhibited stronger binding with Sp1, and then survivin. Subsequent molecular dynamics simulations followed the same trend as initial binding energy calculations and showed crucial amino acids involved in each Mithramycin-A-protein complex. Our findings warrant further investigation into Mithramycin-A and its specific interaction with Sp1 and their downstream targets giving a better understanding of Mithramycin-A and its potential as an effective cancer treatment.

One of the outcomes from the global COVID-19 pandemic caused by SARS-CoV-2 has been an acceleration of development timelines to provide treatments in a timely manner. For example, it has recently been demonstrated that the development of monoclonal antibody therapeutics from vector construction to IND submission can be achieved in five to six months rather than the traditional ten-to-twelve-month timeline using CHO cells [1], [2]. This timeline is predicated on leveraging existing, robust platforms for upstream and downstream processes, analytical methods, and formulation. These platforms also reduce; the requirement for ancillary studies such as cell line stability, or long-term product stability studies. Timeline duration was further reduced by employing a transient cell line for early material supply and using a stable cell pool to manufacture toxicology study materials. The development of non-antibody biologics utilizing traditional biomanufacturing processes in CHO cells within a similar timeline presents additional challenges, such as the lack of platform processes and additional analytical assay development. In this manuscript, we describe the rapid development of a robust and reproducible process for a two-component self-assembling protein nanoparticle vaccine for SARS-CoV-2. Our work has demonstrated a successful academia-industry partnership model that responded to the COVID-19 global pandemic quickly and efficiently and could improve our preparedness for future pandemic threats.

Spike (S) protein, a homotrimeric glycoprotein, is the most important antigen target for SARS-CoV-2 vaccines. A complete simulation of the advanced structure of this homotrimer during subunit vaccine development is the most likely method to improve its immunoprotective effects. In this study, preparation strategies for the S protein receptor-binding domain, S1 region, and ectodomain trimer nanoparticles were designed using ferritin nanoparticle self-assembly technology. The Bombyx mori baculovirus expression system was used to prepare three nanoparticle vaccines with high expression levels recorded in silkworms. The results in mice showed that the nanoparticle vaccine prepared using this strategy could induce immune responses when administered via both the subcutaneous administration and oral routes. Given the stability of these ferritin-based nanoparticle vaccines, an easy-to-use and low-cost oral immunization strategy can be employed in vaccine blind areas attributed to shortages of ultralow-temperature equipment and medical resources in underdeveloped areas. Oral vaccines are also promising candidates for limiting the spread of SARS-CoV-2 in domestic and farmed animals, especially in stray and wild animals.

The pandemic COVID-19 has spread widely throughout the globe and has been responsible for millions of deaths worldwide. Recently, it has been identified that there is no specific and 100% effective treatment available to manage the infection especially for the severe cases. A significant amount of research efforts and clinical trials have been undertaken globally and many more are underway to find the potential treatment option. Earlier, convalescent plasma or hyperimmune immunoglobulin was effectively used in the treatment of many endemic or epidemic viral infections as a part of passive immunization. In this article, we have touched upon the immunopathology of COVID-19 infection, a basic understanding of convalescent plasma, it's manufacturing as well as evaluation, and have reviewed the scientific developments focussing on the potential of convalescent plasma vis-à-vis other modalities for the management of COVID-19. The article also covers various research approaches, clinical trials conducted globally, and the clinical trials which are at various stages for exploring the efficacy and safety of the convalescent plasma therapy (CPT) to predict its future perspective to manage COVID-19.

Leptospirosis is a bacterial disease that affects humans and animals and is caused by . The recommended treatment for leptospirosis is antibiotic therapy, which should be given early in the course of the disease. Despite the use of these antibiotics, their role during the course of the disease is still not completely clear because of the lack of effective clinical trials, particularly for severe cases of the disease. Here, we present the characterization of Lsa45 protein by gel filtration, protein crystallography, SAXS, fluorescence and enzymatic assays. The oligomeric studies revealed that Lsa45 is monomeric in solution. The crystal structure of Lsa45 revealed the presence of two subdomains: a large α/β subdomain and a small α-helical subdomain. The large subdomain contains the amino acids Ser122, Lys125, and Tyr217, which correspond to the catalytic triad that is essential for β-lactamase or serine hydrolase activity in similar enzymes. Additionally, we also confirmed the bifunctional promiscuity of Lsa45, in hydrolyzing both the 4-nitrophenyl acetate (-NPA) and nitrocefin β-lactam antibiotic. Therefore, this study provides novel insights into the structure and function of enzymes from , which furthers our understanding of this bacterium and the development of new therapies for the prevention and treatment of leptospirosis.

The SARS-CoV-2 outbreak and emergence of COVID-19 resulted in the development of different vaccines based on various platforms to combat the disease. While the conventional platforms of inactivated/live attenuated, subunit proteins and virus-like particles (VLPs) have provided efficient and safe vaccines, novel platforms of viral vector- and nucleic acid-based vaccines opened up new horizons for vaccine development. The emergence of COVID-19 pandemic showed that the availability of platforms with high possibility of quick translation from bench to bedside is a prerequisite step in vaccine development in pandemics. Moreover, parallel development of different platforms as well as considering the shipping, storage condition, distribution infrastructure and route of administration are key players for successful and robust response. This review highlights the lessons learned from the current COVID-19 pandemic in terms of vaccine development to provide quick response to future outbreaks of infectious diseases and the importance of vaccine platform in its storage condition and shipping. Finally, the potential application of current COVID-19 vaccine platforms in the treatment of non-infectious diseases has been discussed.

Recent outbreak of COVID-19 pandemic has led to the different possibilities of the development of treatment against corona virus. To know the phylogenicity of SARS-CoV, various studies have been conducted with the outcome of the results showing virulence is caused due to spike protein. Various detection techniques with clinical approach like imaging technology, RT-PCR etc. are comparatively expensively than the use of biosensors. Nano-biosensors have an excellent way of approach to track the conditions of individual and public providing information about the existing condition and treatment status. Electrochemical nano-biosensors are referred as an excellent way of detection. The use of graphene based electrochemical nano-biosensors are most advantageous due to its elevated properties. Fluorescence investigation is one of the precise ways of sensing, optical biosignals that helps in obtaining real time results with high accuracy and negligible changes. The potential application of nano-biosensors are very wide, improvised and advanced Nanotechnology helps in the use of nano-biosensors detect all possible biosignals. Significant ubiquitous IoT-enabled novel sensor technologies that can be potentially utilized to respond various facets the growing COVID-19 pandemic from diagnostic and therapeutics to the prevention stage.

The B.1.617.2 (Delta) variant of concern is causing a new wave of infections in many countries. In order to better understand the changes of the SARS-CoV-2 mutation at the genetic level, we selected six mutations in the S region of the Delta variant compared with the native SARS-CoV-2 and get the conductance information of these six short RNA oligonucleotides groups by construct RNA: DNA hybrids. The electronic characteristics are investigated by the combination of density functional theory and non-equilibrium Green's function formulation with decoherence. We found that conductance is very sensitive to small changes in virus sequence. Among the 6 mutations in the Delta S region, D950N shows the largest change in relative conductance, reaching a surprising 4104.75%. These results provide new insights into the Delta variant from the perspective of its electrical properties. This may be a new method to distinguish virus variation and possess great research prospects.

Cases of deaths due to COVID-19 (COrona VIrus Disease-19) infection are increasing gradually worldwide. Immense research is ongoing to control this pandemic condition. Continual research outcomes are indicating that therapeutic and prophylactic agents are the possible hope to prevent the pandemic from spreading and to combat this increasing death count. Experience gained from previous coronavirus infections (eg., SARS (Severe Acute Respiratory Syndrome), MERS (Middle Ease Respiratory Syndrome), accumulated clinical knowledge during this pandemic, and research helped to identify a few therapeutic agents for emergency treatment of COVID-19. Thereby, monoclonal antibodies, antivirals, broad-spectrum antimicrobials, immunomodulators, and supplements are being suggested for treatment depending on the stage of the disease. These recommended treatments are authorized under medical supervision in emergency conditions only. Urgent need to control the pandemic condition had resulted in various approaches of repurposing the existing drugs, However, poorly designed clinical trials and associated outcomes do not provide enough evidence to fully approve treatments against COVID-19. So far, World Health Organization (WHO) authorized three vaccines as prophylactic against SARS-CoV-2. Here, we discussed about various therapeutic agents, their clinical trials, and limitations of trials for the management of COVID-19. Further, we have also spotlighted different vaccines in research in combating COVID-19.

Amyloidosis refers to a group of diseases caused by the deposition of abnormal proteins in tissues. Herein, curcumin was loaded in a nanohydrogel made of poly (vinylcaprolactam) to improve its solubility and was employed to exert an inhibitory effect on insulin fibrillation, as a protein model. Poly (vinyl caprolactam), cross-linked with polyethylene glycol diacrylate, was synthesized by the reversible addition-fragmentation chain transfer method. The release profile of curcumin exhibited a first-order kinetic model, signifying that the release of curcumin was mainly dominated by diffusion processes. The study of curcumin release showed that 78% of the compound was released within 72 h. The results also revealed a significant decline in insulin fibrillation in the presence of curcumin-loaded poly (vinyl caprolactam). These observations confirmed that increasing the ratio of curcumin-loaded poly (vinyl caprolactam) to insulin concentration would increase the hydrogel's inhibitory effect (P-value < 0.05). Furthermore, transmission electron and fluorescence microscopies and Fourier-transform infrared spectroscopy made it possible to study the size and interaction of fibrils. Based on the results, this nanohydrogel combination could protect the structure of insulin and had a deterrent effect on fibril formation.

The increased infectivity and transmissibility of SARS-CoV-2 new variants were contributed largely by increase binding of receptor binding domain (RBD) domain of the Spike (S) protein to its cellular receptor ACE2 (Angiotensin-Converting Enzyme 2). Several studies have indicated that heparin and its derivatives interact to SARS-CoV-2 S-RBD and inhibits the binding of ACE2 which blocks the viral invasion. However, it is largely unclear how these SARS-CoV-2 variants affects ACE2 binding in the presence of heparin. Herein, using the molecular docking and interaction energy analysis, we showed that N501Y, L452R-E484Q, and E484K mutations bind strongly with heparin in the range of - 7.4 to - 8.0 kcal/mol. The triple mutations, K417N-E484K-N501Y, and K417T-E484K-N501Y displayed weaker binding affinity to heparin (-6.6 kcal/mol). Further, we showed that most of the RBD mutations increased the binding affinity of ACE2 in the absence of heparin, with the maximum increase observed for N501Y (-13.7 kcal/mol). Also, in the presence of heparin, ACE2 binds strongly to the mutant RBD as compared to WT RBD. The strong RBD/ACE2 interaction was observed in case of triple variants (-11.3 kcal/mol) whereas, N501Y showed weakest binding of RBD/ACE2 in the presence of heparin (-9.2 kcal/mol). The strong binding of ACE2 to RBD-heparin complex in these variants will leads to strong inhibition of their entry into host cells.

contains a uronate dehydrogenase termed CsUDH that can convert uronic acids to their corresponding C1,C6-dicarboxy aldaric acids, an important enzyme reaction applicable for biotechnological use of sugar acids. To increase the thermal stability of this enzyme for biotechnological processes, directed evolution using gene family shuffling was applied, and the hits selected from 2-tier screening of a shuffled gene family library contained in total 16 mutations, only some of which when examined individually appreciably increased thermal stability. Most mutations, while having minimal or no effect on thermal stability when tested in isolation, were found to exhibit synergy when combined; CsUDH-inc containing all 16 mutations had Δ +18 °C, such that was unaffected by incubation for 1 hr at ~70 °C. X-ray crystal structure of CsUDH-inc showed tight packing of the mutated residue side-chains, and comparison of rescaled B-values showed no obvious differences between wild type and mutant structures. Activity of CsUDH-inc was severely depressed on glucuronic and galacturonic acids. Combining select combinations of only three mutations resulted in good or comparable activity on these uronic acids, while maintaining some improved thermostability with Δ ~+ 10 °C, indicating potential to further thermally optimize CsUDH for hyperthermophilic reaction environments.

Type III interferons exhibit antiviral activity against influenza viruses, coronaviruses, rotaviruses, and others. In addition, this type of interferon theoretically has therapeutic advantages, in comparison with type I interferons, due to its ability to activate a narrower group of genes in a relatively small group of target cells. Hence, it can elicit more targeted antiviral or immunomodulatory responses. Obtaining biologically-active interferon lambda (hIFN-λ) is fraught with difficulties at the stage of expression in soluble form or, in the case of expression in the form of inclusion bodies, at the stage of refolding. In this work, hIFN-λ was expressed in the form of inclusion bodies, and a simple, effective refolding method was developed. Efficient and scalable methods for chromatographic purification of recombinant hIFN-λ were also developed. High-yield, high-purity product was obtained through optimization of several processes including: recombinant protein expression; metal affinity chromatography; cation exchange chromatography; and an intermediate protein refolding stage. The obtained protein was shown to feature expected specific biological activity in line with published effects: induction of MxA gene expression in A549 cells and antiviral activity against influenza A virus.

Artificial intelligence (AI), a method of simulating the human brain in order to complete tasks in a more effective manner, has had numerous implementations in fields from manufacturing sectors to digital electronics. Despite the potential of AI, it may be obstinate to assume that the person administered society would rely solely on AI; with an example being the healthcare field. With the ever-expanding discoveries made on a regular basis regarding the growth of various diseases and its preservations, utilizing brain power may be deemed essential, but that doesn't leave AI as a redundant asset. With the years of accumulated data regarding patterns and the analysis of various medical circumstances, algorithms can be formed, which could further assist in situations such as diagnosis support and population health management. This matter becomes even more relevant in today's society with the currently ongoing COVID-19 pandemic by SARS-CoV-2. With the uncertainty of this pandemic from strain variants to the rolling speeds of vaccines, AI could be utilized to our advantage in order to assist us with the fight against COVID-19. This review briefly discusses the application of AI in the COVID-19 situation for various health benefits.

The severe acute respiratory syndrome coronavirus 2, famous as COVID-19, has recently emerged as a novel virus and imposed an unrecoverable loss to global health and the economy. At present, no effective drug against COVID-19 is available and currently available viral drugs targeting the viral key proteins of related RNA viruses have been found ineffective against COVID-19. This study evaluated the inhibitors of the viral proteases and other structural proteins, including Mpro (Main protease), RdRp (RNA-dependent RNA polymerase), and spike glycoprotein from synthetic and herbal sources. The molecular docking-based approach was used to identify and evaluate the putative inhibitors of key proteins involved in viral replication and survival. Furthermore, the pharmaceutical properties of these inhibitors were explored to predict the drug suitability as a therapeutic agent against COVID-19 by considering adsorption, distribution, metabolism, and excretion (ADME) using Lipinski's rule or SwissADME. Trandolapril, Benazepril, and Moexipril were evaluated as the best non-carcinogenic and non-toxic potential inhibitors of spike glycoprotein, Mpro, and RdRp, respectively. The drugs showed significant binding affinities against the active sites of respective SARS_CoV-2 target proteins; hence, they can be used as potential therapeutic agents for the treatment of COVID-19.

To tackle the harmful consequences of the widespread COVID-19 pandemic, a broad-spectrum anti-viral drug remdesivir (RDV) has gained the utmost attention recently due to its promising application in treating COVID-19 patients. However, a fast and sensitive analytical methodology is important to monitor RDV drug profile in human plasma for pharmacokinetics (PK) and therapeutic drug monitoring (TDM). In this study, we demonstrate an improved vortex-assisted salt-induced liquid-liquid microextraction (VA-SI-LLME) technique coupled with UHPLC-PDA and UHPLC-MS/MS for rapid determination of RDV in human plasma. This technique involves simple one-step protein precipitation with hydrochloric acid and subsequent extraction with acetonitrile for analysis. Under the optimal VA-SI-LLME conditions (500 μL of acetonitrile with 2.5 g ammonium sulfate under 2 min vortex extraction), method validation results indicated an excellent correlation coefficient of 0.9969 for UHPLC-PDA (monitored at 254 nm) and 0.9990 for UHPLC-MS/MS (monitored at electrospray ionization with + ion mode transitions of 603.1→ 402.20 and 603.1→ 199.90). The detection and quantification limits were 1.5 and 5 ng/mL for UHPLC/PDA, and 0.3 and 1 ng/mL for UHPLC-MS/MS, respectively. The developed method showed excellent extraction recoveries between 90.79-116.74 % and 85.68-101.34 % with intraday and interday precision ≤ 9.59 for both methods. These results proved that the developed method is a simple, fast, and sensitive analytical method that can be applied as a standard analytical tool for PK and TDM studies of RDV in clinical trials during the current worldwide outbreak.

Here we describe refined methods for the isolation and detection of a KDEL-tagged, plant-produced recombinant cholera toxin B subunit (CTB) that exhibits unique mucosal wound healing activity. The protein was transiently overexpressed in , which generates some C-terminal KDEL truncated molecular species that are deficient in epithelial repair activity. With a new CHT chromatographical method described herein, these product-derived impurities were successfully separated from CTB with the intact KDEL sequence, as confirmed by mass spectrometry. In addition, an immunoassay capable of specifically detecting GM1 ganglioside-binding CTB with intact KDEL sequences was developed. Coupled together, these methods will aid in the quality control of KDEL-attached CTB produced in plant-based manufacturing systems towards a novel topical biotherapeutic for the treatment of acute and chronic mucosal inflammation.

Microbial fuel cells (MFCs) offer an excellent solution to tackle some of the major challenges currently faced by humankind: sustainable energy sources, waste management and water stress. Besides treating wastewater and producing useful electricity from urine, ceramic MFCs can also generate biocidal catholyte . It has been proved that the electricity generation from the MFCs has a high impact in the catholyte composition. Therefore, the catholyte composition constantly changes while electricity is generated. However, these changes in catholyte composition with time has not yet been studied and that could highly contribute to the disinfection efficacy. In this work, the evolution of the catholyte generation and composition with the MFC operation time has been chemically and microbiologically evaluated, during 42 days. The results show an increase in pH and conductivity with the operation time, reaching pH 11.5. Flow cytometry and luminometer analyses of bioluminescent pathogenic exposed to the synthesised catholyte revealed killing properties against bacterial cells. A bio-electrochemical system, capable of electricity generation and simultaneous production of bactericidal catholyte from human urine is presented. The possibility to electrochemically generate a bacterial killing agent from urine, offers a great opportunity for water reuse and resource recovery for practical implementations.

Nanomaterials have wide-ranging biomedical applications in prevention, treatment and control of diseases. Nanoparticle based vaccines have proven prodigious prophylaxis of various infectious and non-infectious diseases of human and animal concern. Nano-vaccines outnumber the conventional vaccines by virtue of plasticity in physio-chemical properties and ease of administration. The efficacy of nano-based vaccines may be attributed to the improved antigen stability, minimum immuno-toxicity, sustained release, enhanced immunogenicity and the flexibility of physical features of nanoparticles. Based on these, the nano-based vaccines have potential to evoke both cellular and humoral immune responses. Targeted and highly specific immunological pathways required for solid and long lasting immunity may be achieved with specially engineered nano-vaccines. This review presents an insight into the prevention of infectious diseases (of bacterial, viral and parasitic origin) and non-infectious diseases (cancer, auto-immune diseases) using nano-vaccinology. Additionally, key challenges to the effective utilization of nano-vaccines from bench to clinical settings have been highlighted as research domains for future.

generation of red blood cells (RBCs) from hematopoietic stem cells (HSCs) used for blood transfusion represents one of the focuses in current regenerative medicine. However, massive production of HSCs-based RBCs requires a significant quantity of erythropoietic growth factors, making manufacturing at large scale cost prohibitive. Plant cell culture is proposed to be a promising bioproduction platform for functional human proteins in a safe and cost-efficient manner. This study exploited a proprietary technology, named HypGP engineering technology, for high-yield production of one of the key erythropoietic growth factors--stem cell factor (SCF)--in plant cell culture. Specifically, a designer hydroxyproline (Hyp)--glycosylated peptide (HypGP) comprised of 20 tandem repeats of the "Ser-Pro" motif, or (SP) was engineered at either the N-terminus or C-terminus of SCF in tobacco BY-2 cells. The (SP) tag dramatically increased the secreted yields of SCF up to 2.5 μg/ml. The (SP)-tagged SCF showed bioactivity in promoting the proliferation of the TF-1 cell line, although the SCF-(SP) was 8.4-fold more potent than the (SP)-SCF. Both the (SP)-SCF and SCF-(SP) exhibited desired function in stimulating the expansion and differentiation of human umbilical cord blood CD34 cells towards RBCs.

As the new cases of COVID-19 are growing every daysince January 2020, the major way to control the spread wasthrough early diagnosis. Prevention and early diagnosis are the key strategies followed by most countries. This study presents the perspective of different modes of transmission of coronavirus,especially during clinical practices and among the pediatrics. Further, the diagnostic methods and the advancement of the computerized tomography have been discussed. Droplets, aerosol, and close contact are thesignificantfactors to transfer the infection to the suspect. This study predicts the possible transmission of the virus through medical practices such as ophthalmology, dental, and endoscopy procedures. With regard to pediatric transmission, as of now, only afew child fatalities had been reported. Childrenusually respond to the respiratory virus; however, COVID-19 response ison the contrary. The possibility of getting infected is minimal for the newborn. There has been no asymptomatic spread in children until now. Moreover, breastfeedingwould not transmit COVID-19, which is encouraging hygiene news for the pediatric. In addition, the current diagnostic methods for COVID-19 including Immunoglobulin M (IgM) and Immunoglobulin G (IgG)and chest computed topography(CT) scan, reverse transcription-polymerase chain reaction (RT-PCR) andimmunochromatographic fluorescence assay, are also discussed in detail. The introduction of artificial intelligence and deep learning algorithmhas the ability to diagnose COVID-19 in precise. However, the developments of a potential technology for the identification of the infection, such as a drone with thermal screening without human intervention, need to be encouraged.