Automated Sensor Node Malicious Activity Detection with Explainability Analysis.

Cybersecurity has become a major concern in the modern world due to our heavy reliance on cyber systems. Advanced automated systems utilize many sensors for intelligent decision-making, and any malicious activity of these sensors could potentially lead to a system-wide collapse. To ensure safety and security, it is essential to have a reliable system that can automatically detect and prevent any malicious activity, and modern detection systems are created based on machine learning (ML) models. Most often, the dataset generated from the sensor node for detecting malicious activity is highly imbalanced because the Malicious class is significantly fewer than the Non-Malicious class. To address these issues, we proposed a hybrid data balancing technique in combination with a Cluster-based Under Sampling and Synthetic Minority Oversampling Technique (SMOTE). We have also proposed an ensemble machine learning model that outperforms other standard ML models, achieving 99.7% accuracy. Additionally, we have identified the critical features that pose security risks to the sensor nodes with extensive explainability analysis of our proposed machine learning model. In brief, we have explored a hybrid data balancing method, developed a robust ensemble machine learning model for detecting malicious sensor nodes, and conducted a thorough analysis of the model's explainability.

Novel Coronavirus: A Review from Origin to Current Status of Therapeutic Strategies.

Coronaviruses (CoVs) are continuously emerging, highly transmissible, and pathogenic agents that primarily target the human respiratory system. Previous outbreaks of severe acute respiratory syndrome-CoV and Middle East respiratory syndrome-CoV remain life-threatening and global public health concerns. A novel CoV outbreak that occurred in December 2019 in Wuhan, China was declared a pandemic outbreak that has since killed millions of individuals worldwide. Rapid transmission, genetic variations, and unavailability of specific therapeutic drugs are major factors that led to this alarming and deadly situation. Currently, > 200 clinical vaccine trials are underway to combat infection. This review summarizes reports related to CoV origin, genetic variations, drug options, status of nine vaccines that were in phase III trials, and novel therapies including convalescent plasma and stem cell treatment.

Dengue Vaccines: Ongoing Challenges and Current Status in the Advancement of Different Candidates.

Dengue is a vector-borne highly systemic infectious disease of the tropical and subtropical countries and is devastating millions of lives worldwide. It may be self-eliminated like a mild fever or may cause life-threatening fatal complications as dengue hemorrhagic fever and dengue shock syndrome. The lack of specific and effective antiviral drugs and vaccines amplify its transmission rate across the world. The development of the dengue vaccine has been an ambitious task due to the presence of four different dengue serotypes capable of carrying antibody enhancement complex mechanisms. In this review, we have summarized the ongoing challenges in the construction of a dengue vaccine and the current status of the vaccine development. Limited knowledge of immune responses against dengue infection, lack of human or animal model of disease, and suboptimal assay strategies to detect immune responses after infection or vaccination, are some barriers to vaccine and drug development. A tetravalent vaccine with low cost, high efficiency, and capable of eliciting immune responses against all four serotypes is needed to minimize the epidemics. Currently, only one live attenuated chimeric dengue vaccine, the CYD Dengue Vaccine, has completed its third phase and has been licensed. DENVax and TetraVax-DV-TV003 (TV003) are in the third phase while others are still in the first trial phase.

Sanguinarine disrupts the colocalization and interaction of HIF-1α with tyrosine and serine phosphorylated-STAT3 in breast cancer.

Breast cancer is one leading cause of death in females, especially triple-negative breast cancer (TNBC). Hypoxia is a key feature leading to tumour progression driven by hypoxia-inducible factor (HIF)-1α. The aim is to investigate the mechanism of HIF-1α and signal transducer and activator of transcription-3 (STAT3) interaction and discover a compound to disrupt the interaction in breast cancer cells. The regulation pattern of HIF-1α and STAT3 was analysed in hypoxic TNBC cells and patient samples. The effects of a natural alkaloid, sanguinarine, on HIF-1α and STAT3 colocalization and interaction were evaluated in vitro and mouse xenograft models. We observed strong colocalization of HIF-1α, p-STAT3-Tyr and p-STAT3-Ser in TNBC patient samples. Sanguinarine could inhibit the nuclear colocalization and interaction of HIF-1α with p-STAT3-Tyr and p-STAT3-Ser in vivo and in vitro. Our results may bring insights to the HIF-1α/STAT3 interaction in breast cancers and suggest sanguinarine as a promising candidate for HIF-α/STAT3 inhibition.

Production of potent long-lasting consensus interferon using albumin fusion technology in Pichia pastoris expression system.

Consensus interferon (cIFN) is a wholly synthetic therapeutic protein which is used to treat hepatitis C/B and certain types of malignancies. It has short serum half-life, therefore, to maintain its therapeutic level in the human body it requires thrice-weekly administration. Various strategies like PEGylation and micro-encapsulation have been developed during the last few years to enhance the pharmacokinetics of small therapeutic peptides. This study executed the human albumin-fusion technology, a simple and flexible approach to extend the serum circulating half-life of cIFN, because human serum albumin (HSA) has long circulating half-life (19 days) and very minute immunological activities. We integrated the codon-optimized HSA-cIFN fusion gene into Pichia pastoris genome by homologous recombination. The selection of hyper-resistant P. pastoris clone against Zeocin™ achieved a high-level secretory expression (250 mg/L) of fusion protein. HSA-cIFN fusion protein was purified using one-step purification by affinity chromatography with 34% recovery. The SDS-PAGE and SEC-HPLC analysis confirmed the final purified product has molecular weight of 87 kDa with 98% purity. Western blot analysis using anti-IFN antibodies further verified the purified HSA-cIFN fusion protein. The specific biological activity was 2.1 × 106 IU/mg as assessed by cytopathic inhibition assay, and half-life of fusion protein was estimated by in vitro thermal and proteolytic stability studies. This work concludes that by using albumin fusion technology, codon optimization and one-step purification a high yield of 86 mg/L of biologically active protein with improved serum half-life was obtained.

Mechanistic insight into the electrocatalytic performance of reduced graphene oxide supported palladium, silver and palladium-silver nanodeposits toward electro-dehalogenation of halocarbons in room temperature ionic liquids.

Herein, we report the results from our extensive voltammetric investigations designed to explore, assess and explain the electrocatalytic performance of reduced graphene oxide supported metal nano-deposits toward the electro-dehalogenation of halocarbons in room temperature ionic liquids (RTILs). Specifically, we investigated the electro-reductive dechlorination of the model halocarbon, carbon tetrachloride over glassy carbon electrode (GCE) and palladium-graphene (Pd-Gr), silver-graphene (Ag-Gr) and palladium-silver-graphene (PdAg-Gr) nanocomposites in 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([BMIM][NTf2]). Analysis of the voltammetric data in light of Marcus-Hush formulation reveals that the electro-reductive cleavage of the C-Cl bond of CCl4 over GCE in [BMIM][NTf2] follows a sticky dissociative electron transfer (SDET) pathway. The significantly stronger interaction energy between electrogenerated Cl- and CCl3˙ (radical) fragments in RTILs makes electroreduction of CCl4 in [BMIM][NTf2] much easier than in organic solvents. The activation-driving force relationship for electro-catalytic dechlorination of CCl4 over Pd-Gr was observed to follow a modified sticky dissociative electron transfer model wherein apart from the ion-radical interaction, the adsorptive interaction of chlorinated species with the electrocatalytic surface needs to be taken into consideration to account for the apparent activation energy-driving force dependence. Interestingly the activation energy-driving force relationships for the electroreduction of CCl4 over Ag-Gr and PdAg-Gr were observed to fit a modified stepwise ET (MSET) pathway. In the MSET pathway, the adsorption and the implied free energy change of the electroreducible halocarbon significantly alter the solvent re-organization energy and the inherent barrier for the heterogeneous ET process. The adsorptive interaction and hence the electrocatalytic performance of PdAg-Gr were observed to be more than that observed for Ag-Gr. This is attributed to the Ag to Pd charge transfer in the PdAg-Gr nanodeposits. Our results besides underlining the positive influence of RTILs in facilitating the electroreductive detoxification of halocarbons, very well establish the mechanistic basis for the electrocatalytic performance of graphene based nanodeposits toward electrodehalogenation of halocarbons.

Transforming micelles into mixed micelles: a promising approach to tune the catalytic performance of imidazolium-based surface active ionic liquids toward degradation of rhodamine B.

Herein, we demonstrate that the catalytic performance of imidazolium-based surface-active ionic liquid (SAIL) micelles can be significantly enhanced through the addition of an appropriate type and amount of intelligently conceived amphiphile to form mixed micelles. Specifically, we show that the catalytic performance of 1-dodecyl-3-methyl imidazolium chloride (DDMIMCl) micelles toward the reductive degradation of rhodamine B (RhB), a carcinogenic dye extensively used in multiple industrial applications, can be appreciably boosted through addition of Brij56, a nonionic surfactant. Detailed kinetic investigations on the catalytic performance of pre- and post-micellar concentrations of DDMIMCl and its mixed micelles with Brij56 over various mole fractions, toward the reductive degradation of RhB, are presented. The data analyzed in light of Berezin's kinetic model suggest that the addition of Brij56 to DDMIMCl micelles significantly enhances their catalytic performance. The catalytic activity exhibited by the DDMIMCl-Brij56 (XBrij56 = 0.2) mixed micellar system is better than that reported for many state-of-the-art nanoparticle/homogenous catalysts. The results explained in light of Berezin's kinetic model are well supported by physico-chemical studies like conductometry, fluorimetry and dynamic light scattering. The presented results anticipate stimulation of extensive research activity for exploiting the mixed micellization approach as a novel avenue for modulating the catalytic performance of SAILs.

Evaluating the ion exchange chromatography for matrix-assisted PEGylation and purification of consensus interferon.

Scientists have implemented protein-PEGylation technology for boosting-up the pharmacokinetics and stability of recombinant therapeutic proteins. In the present study, (a) matrix-assisted PEGylation was compared with solution-phase PEGylation and (b) matrix-assisted PEGylation was performed with different ion exchange resins for impact of chromatography medium on yield and purity of PEGylated product. DEAE Sepharose CL 6B, DEAE Fracto gel, and Macro cap Q ion exchange chromatography medium were compared for on column PEGylation and purification of cIFN. A MSC-PEG of 12.0 KDa was selected. cIFN was bound to ion exchange medium, and PEG solution was passed through resin for 180 Min, and protein was eluted by sodium chloride linear gradient. Yield and purity for mono-PEGylated cIFN with Macro cap Q matrix was 75% and 99%, respectively, whereas for DEAE Sepharose was 45% and 60%. DEAE Fracto gelTM purity was 85% with 50% yield of mono-PEGylated cIFN. Further investigation of in vitro biological activities demonstrated that about 30% antiviral activity was reduced as compared to unmodified cIFN. However, thermal stability was significantly improved. The present study proved that matrix-assisted PEGylation can improve the yield and purity of mono-PEGylated product, and Macro Cap resin provided the highest yield of a homogeneous product. In present study, (a) matrix-assisted PEGylation was compared with solution-phase PEGylation and (b) matrix-assisted PEGylation was performed with different ion exchange resins for impact of chromatography medium on yield and purity of PEGylated product. Matrix-assisted PEGylation increases the yield of mono-PEGylated product and further Macro CapTM produced highest yield and purity of PEGylated cIFN.

Enhancing recombinant interleukin-6 production yield by fermentation optimization, two-step denaturing, and one-step purification.

Interleukin-6 a pleiotropic cytokine involved in a wide range of biological activities. So the large-scale production of biologically active recombinant human interleukin-6 is important for its structural and functional studies. Here, we report an optimized method for shake flask fermentation and a simplified high-yield purification procedure for the recombinant interleukin-6. This high-yield expression method not only involves the optimization of the fermentation condition but also the single step purification method as well as a two-step denaturing and one-step refolding process. This approach replaces the more conventional procedure of protein solubilization and refolding. Through applying these strategies, the final cell density and overall product yield of the recombinant human interleukin-6 were obtained as 20.4 g as cell biomass and 150 mg as purified active protein from the I-L of the culture. The purified protein was characterized by HPLC and SDS-PAGE. The results of the current work demonstrate that the described method may be used to develop the process for industrial-scale production of the biologically active recombinant interleukin-6 protein.

Towards Understanding the Solvent-Dynamic Control of the Transport and Heterogeneous Electron-Transfer Processes in Ionic Liquids.

The impact of temperature-induced changes in solvent dynamics on the diffusion coefficient and standard rate constant k0 for heterogeneous electron transfer (ET) of ethylferrocene (EFc) in 1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM][PF6 ]) is investigated. The results are analysed to understand the impact of solvent-dynamic control, solute-solvent interactions and solvent friction on the transport of redox probes and k0 . Concentration dependence of the diffusion coefficient of EFc in [BMIM][PF6 ] is observed. This is attributed to the solute-induced enhancement of the structural organisation of the ionic liquid (IL), which is supported by the concentration-dependent UV/Vis absorption and photoluminescence responses of EFc/[BMIM][PF6 ] solutions. Similar values of the activation energies for mass transport and ET and a linear relationship between the diffusion coefficient and the heterogeneous ET rate is observed. The ratio between the diffusion coefficient and the heterogeneous rate constant allows a characteristic length Ld , which is temperature-independent, to be introduced. The presented results clearly establish that mass transport and heterogeneous ET of redox probes are strongly correlated in ILs. It is proposed that the apparent kinetics of heterogeneous ET reactions in ILs can be explained in terms of their impact on thermal equilibration, energy dissipation and thermal excitation of redox-active probes.

Evaluating the autoinduction expression system and one-step purification for high-level expression and purification of gallbladder-derived rhIL-1Ra.

Recent advancement in fermentation technologies resulted in the increased yields of recombinant proteins of biopharmaceutical and medicinal importance. Consequently, there is an important task to develop simple and easily scalable methods that can facilitate the production of high-quality recombinant protein. Most of the recent reports described the expression of recombinant human IL-1 receptor antagonist (rhIL-1Ra) in Escherichia coli using isopropyl-ß-d-thiogalacto pyranoside (IPTG), a nonmetabolizable and expensive compound, as an expression inducer. In this study, we describe the expression and one-step purification of gallbladder-derived rhIL-1Ra by autoinduction in E. coli. This method includes special media that automatically induce the target protein expression from T7 promoter and allow the production of the target protein in high yield than the conventional IPTG induction method. In addition to fermentation process improvements, one-step purification strategy is essential to make the process economical. We developed a single-step cation exchange chromatography and obtained 300 mg/L of rhIL-1Ra with 98% purity. Purified protein was characterized by SDS-PAGE and Ion exchange HPLC (IEX-HPLC). The described method can be used to scale up the production of rhIL-1Ra and other recombinant proteins.

Bioactivity studies of Huh-7 cells derived human epidermal growth factor expressed in Pichia pastoris.

Previously, we have reported cloning of human epidermal growth factor gene from Huh-7 cells and its extracellular expression in Pichia pastoris. The presented work is a detailed report regarding molecular characterization of Huh-7 cells-derived hEGF expressed in Pichia pastoris with special reference to its glycosylation profiling and bioactivity studies. Densitometric scanning of SDS-PAGE separated extracellular proteins from hEGF recombinant Pichia pastoris strain indicated that about 84% of the extracellular proteins were glycosylated. Size exclusion chromatography using Superdex 75 prep grade column was successfully utilized to separate fractions containing glycosylated and non-glycosylated extracellular proteins. In dot blot assay, hEGF was detected in both glycosylated and non-glycosylated fractions. Bioactivity assays revealed that both glycosylated and non-glycosylated fractions were bioactive as determined by cell viability assay. It was also observed that hEGF present in non-glycosylated fraction was relatively more bioactive than hEGF present in glycosylated fraction.

Simple procedure applying lactose induction and one-step purification for high-yield production of rhCIFN.

Recombinant consensus interferon (CIFN) is a therapeutic protein with molecular weight of 19.5 kDa having broad spectrum antiviral activity. Recombinant human CIFN (rhCIFN) has previously been expressed in Escherichia coli using isopropyl-ß-d-thiogalactopyranoside (IPTG), a non-metabolizable and expensive compound, as inducer. For economical and commercial-scale recombinant protein production, it is greatly needed to increase the product yield in a limited time frame to reduce the processing cost. To reduce the cost of production of rhCIFN in E. coli, induction was accomplished by using lactose instead of IPTG. Lactose induction (14 g/L) in shake flask experiment resulted in higher yield as compared with 1 mM IPTG. Finally, with single-step purification on DEAE sepharose, 150 mg/L of >98% pure rhCIFN was achieved. In the present study, an attempt was made to develop a low cost process for producing quality product with high purity. Methods devised may be helpful for pilot-scale production of recombinant proteins at low cost.

Oxides in silver-graphene nanocomposites: electrochemical signatures and electrocatalytic implications.

Herein we report an electrochemical approach to establish the presence of silver oxides in silver-reduced graphene oxide (Ag-rGO) nanocomposites synthesised under alkaline conditions. The recorded electrochemical signatures, further supported and validated by UV-Vis spectroscopy, XRD and TEM analysis, clearly establish the presence of an oxide phase of silver in the nanodimensional silver present in Ag-rGO. The Ag-rGO was tested for its electrocatalytic and electrosensing activity for hydroquinone (H2Q) and ascorbic acid (AA). The presented results establish that the electrocatalytic and electrosensing potential of the Ag-rGO for H2Q and AA can be enhanced through electroreduction of the oxide phase of silver in these nanocomposites. Our results prove that the electrocatalytic and electroanalytic activities of electroreduced Ag-rGO for AA are better than most of the electrode materials reported so far in the literature.

Optimization of conditions for high-level expression and purification of human recombinant consensus interferon (rh-cIFN) and its characterization.

Recombinant human consensus interferon (rh-cIFN) is an artificially engineered interferon (IFN) developed by recombining and reordering the protein sequences that exist in standard IFN. This recombination resulted into a drug that has the potential to work better than natural, standard IFN. In this study, we described optimized conditions for high-level expression and recovery of biologically active consensus IFN from inclusion bodies (IBs). A synthetic gene coding 166 amino acids of consensus IFN was cloned under the T7 promoter. Escherichia coli strain BL21DE3Plys was used to transform expression construct. For high-level expression, shake-flask fermentation conditions were standardized. For isolation of IBs, the sonication method was optimized. A variety of chaotropic agents including guanidine hydrochloride, urea, SDS, and detergents were studied for solubilization of IBs. For renaturation of solubilized denatured protein by the dilution process, parameters of dilution factor, temperature, and l-arginine were optimized. A one-step chromatography method was developed for high-yield purification of consensus IFN. rh-cIFN was characterized by SDS-PAGE, Western blot, and high-performance liquid chromatography. Purified protein has a molecular weight of 19.5 kDa and specific activity was 2.0 × 10(8) as determined by the cytopathic inhibition assay. This study concludes that by using optimized conditions, we obtained a yield of 100 mg/L of biologically active rh-cIFN, which is highest ever reported according to available data.

Studies to analyse the relationship between IFNα2b gene dosage and its expression, using a Pichia pastoris-based expression system.

Human interferon α2b (hIFNα2b) is the most important member of the interferon family. Escherichia coli, yeasts, mammalian cell cultures and baculovirus-infected insect cells have been used for expressing recombinant human interferon. Recently a Pichia pastoris-based expression system has emerged as an attractive system for producing functional human recombinant IFNα2b. In this regard, gene dosage is considered an important factor in obtaining the optimum expression of recombinant protein, which may vary from one protein to another. In the present study we have shown the effect of IFNα2b gene dosage on extracellular expression of IFNα2b recombinant protein from P. pastoris. Constructs containing from one to five repeats of IFNα2b-expressing cassettes were created via an in vitro multimerization approach. P. pastoris host strain X-33 was transformed using these expression cassettes. Groups of P. pastoris clones transformed with different copies of the IFNα2b expression cassette were screened for intrachromosomal integration. The IFNα2b expression level of stable transformants was checked. The copy number of integrated IFNα2b was determined by performing qPCR of genomic DNA of recombinant P. patoris clones. It was observed that an increase in copy number generally had a positive effect on the expression level of IFNα2b protein. Regarding the performance of multicopy strains, those obtained from transformation of multicopy vectors showed relatively high expression, compared to those generated using transformation vector having only one copy of IFNα2b. It was also observed that an increase in drug resistance of a clone did not guarantee its high expression, as integration of a marker gene did not always correlate with integration of the gene of interest.

Expression line approach to recombinant human epidermal growth factor into the yeast, Pichia pastoris from Huh-7 cell line.

Beta-urogastrone also known as human epidermal growth factor is a key member of epidermal growth factor family having role in cell proliferation and differentiation in vivo as well as in vitro. Human epidermal growth factor gene has been isolated from different tissues but the method of isolation is technically difficult and complicated as it deals with biopsies. Here we isolated mature partial human epidermal growth factor gene from Huh-7 cell line, amplified and abridged toward mature coding region with three steps PCR, sequenced for homology with wild type human epidermal growth factor gene, inbuilt with sites of interest and cloned in Pichia pastoris for expression study. Isolated mature human epidermal growth factor gene from Huh-7 cell line showed 100 % sequence homology to wild type human epidermal growth factor gene and gives the native expression for human epidermal growth factor peptide. In this study we report that Huh-7 cell line is an easy source for the particular gene of human epidermal growth factor isolation and we are also suggesting P. pastoris is an expression system to produce recombinant human epidermal growth factor of the therapeutic importance resembling to the natural human system.

Matrix-assisted refolding and purification of placenta-derived recombinant human interleukin-6 produced in Escherichia coli.

Biological activity of human interleukin-6 (IL-6) is associated with a vast number of diseases such as rheumatoid arthritis, sepsis, and severe inflammatory diseases. In this study, human IL-6 cDNA was isolated from a cDNA library that was constructed with mRNA derived from human placental tissues. Subsequently, the complete human IL-6 cDNA was cloned and expressed in BL21DE3 cells. The recombinant human IL-6 (rhIL-6) protein was expressed in a form of an insoluble inclusion body. Inclusion bodies were solubilized under denaturing conditions and purified by immobilized metal affinity chromatography with gradual on-column refolding by the gradient elution method (from 6 to 0 M urea). The protein was purified to apparent homogeneity of about 99% with a yield of 50 mg/L. The purity was assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), size exclusion high-performance liquid chromatography, and Western blotting analysis. The bioactivity was assessed by proliferation assay of TF-1 cells in a dose-dependent manner. The present study confirms the expression of the placenta-derived IL-6 gene in a prokaryotic expression system and matrix-assisted on-column refolding and purification of rhIL-6 by immobilized metal affinity chromatography.

Replacing imidazole with benz-imidazole: a promising approach to enhance the electrocatalytic performance of imidazolium based surface active ionic liquids for greener electrosynthesis.

We present the synthesis of a novel benzimidazolium-based surface active ionic liquid (SAIL), 3-dodecyl-1H-3λ-benzo[d]imidazole chloride, a SAIL with excellent surface activity and self-aggregation tendency whose aqueous micellar solutions offer exceptional solubilizing capacity and electrocatalytic performance for efficient electrocarboxylation of halocarbons.

An oral toxicity assessment of a mosquito larvicidal transgenic algae (Chlamydomonas reinhardtii) using adult Zebrafish and its embryos.

Mosquito-borne diseases pose a global health threat, with pathogens like Malaria, Dengue fever, and others transmitted by mosquitoes. Our study focuses on evaluating the toxicity of genetically engineered mosquito larvicidal algae (Chlamydomonas reinhardtii) to non-target organisms, specifically Zebrafish. We conducted a 90-day experiment, feeding Zebrafish different combinations of larvicidal algae and commercial fish feed. Statistical analysis revealed no significant differences in mortality, allergenicity, or moribundity among groups. Hematology, molecular analysis, and necropsy showed no physiological differences. Our findings indicate that the transgenic algae (TN72.cry11Ba) had no adverse effects on adult Zebrafish or their larvae. This study confirmed the safety of algae on non-target organisms, such as zebrafish.