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In this era of technology, data storage and its management plays an important role. Every organization include loads of data, storing and managing bundle of data is a major issue. A convenient approach is required to handle it so that accessing and updating it become simple and easy. Apart from storage and management, freelancing is an emerging trend nowadays. It requires programming languages and their frameworks to cater to the consumer interests. Yet the availability of all frameworks on a single system is an arduous task which can be eradicated by creating our own database for the purpose of data storage, multiple system usage and software development. But it is costly and time-consuming task. For proper utilization of time, money and space, Cloud Computing comes into play these days. Therefore, in this module, we will discuss about Cloud Computing its working, deployment and service mode of Cloud Computing along with their pros and cons.

Huanglongbing (HLB) is a worldwide citrus plant disease-related to non-culturable and fastidious α-proteobacteria Candidatus Liberibacter asiaticus (CLas). In CLas, Peroxiredoxin (Prx) plays a major role in the reduction of the level of reactive species such as reactive oxygen species (ROS), free radicals and peroxides, etc. Here, we have used structure-based drug designing approach was used to screen and identify the potent molecules against 2Cys Prx. The virtual screening of fragments library was performed against the three-dimensional validated model of Prx. To evaluate the binding affinity, the top four molecules (N-Boc-2-amino isobutyric acid (B2AI), BOC-L-Valine (BLV), 1-(boc-amino) cyclobutane carboxylic acid (1BAC), and N-Benzoyl-DL-alanine (BDLA)) were docked at the active site of Prx. The molecular docking results revealed that all the identified molecules had a higher binding affinity than Tert butyl hydroperoxide (TBHP), a substrate of Prx. Molecular dynamics analysis such as RMSD, Rg, SASA, hydrogen bonds, and PCA results indicated that Prx-inhibitor(s) complexes had lesser fluctuations and were more stable and compact than Prx-TBHP complex. MMPBSA results confirmed that the identified compounds could bind at the active site of Prx to form a lower energy Prx-inhibitor(s) complex than Prx-TBHP complex. The identified potent molecules may pave the path for the development of antimicrobial agents against CLA. Communicated by Ramaswamy H. Sarma

The amount of raw data has exponentially increased due to tools and technologies for analysis of biologic organisms (including viruses). The task now, therefore, is to have computational systems that allow the average researcher to calculate information to store, retrieve, visualize, and analyze data. Fortunately, the advancement of computer technology, particularly that relating to databases, has balanced continuous improvements to data-intensive biologic technologies. This research forms the foundation of many of the bioinformatics technologies that provide an overview of the different databases available for viruses and how they provide valuable information from the stored biologic data. The different tools associated with virus analysis are also discussed, accompanied by a summary of several online databases that contain general biologic and viral data. The chapter also features a case study on evaluating genome relationship between SARS-CoV-2 strains at different geolocations.

The COVID-19 pandemic has brought into further crisis the contemporary global university. By focusing on the UK this article examines how this crisis has manifested itself in UK universities, with ...

In this paper, an artificial human interface based on magnetic resonance coupling is proposed to detect and sense the direction of movement of a fingertip. The finger movement is analyzed by recognizing the change in magnetic coupling observed between a static transmitter coil and a static receiver coil array due to presence of a small coil resonator placed at the fingertip. By measuring induce voltage responses in the spatially distributed receiver coil array, the movement of the resonator coil is characterized. A commercial software Ansys EM suite 19.1 is utilized to demonstrate applicability of the proposed design. The fingertip movement is examined by moving the resonator in various directions and the observed results corroborate the claims. Therefore, the proposed touchless pad is found well suitable for infection free accessibility of the computer systems.

In this paper, a novel trapezoidal multi-coil antenna is proposed to mitigate lateral misalignment in wireless charging application of electric vehicles (EVs). The proposed coil antenna used at the transmitter is optimized to improve stability of induced voltage which intern is expected to provide a stable power transfer efficiency over a wide range of vehicle’s misalignment. The proposed design adopts an isosceles trapezoid shape after analytical optimization and consists of three series connected coils placed at the top faces of the structure. The magnetic field distribution originated from this complex geometry is analytically modeled and optimized to obtain wider uniformity in the induced voltage of the receiver coil as compared to a conventional planar antenna. The design is verified using commercial EM software and results are corroborated. The results show that the proposed antenna provides enhanced tolerance towards misalignment is proved to be a potential design for wireless powering application of EVs.

A compact wideband microstrip antenna operating over 5.45 GHz - 7.47 GHz bandwidth is reported. The design uses double rectangular type annular-ring structured patch, along with a circular arc at each corner acting as defected metal structure (DMS) and two triangular-shaped defects in the ground plane acting as defected ground structures (DGS). The annular ring contributes to increase bandwidth of basic microstrip antenna and further enhancement in bandwidth is achieved by combination of both DMS and DGS. The design is implemented on a RT/Duroid 5880 TM substrate. The overall design size is 14 mm×18 mm. The maximum simulated gain of the proposed antenna is 7.8 dBi. Hence, the proposed design is found suitable for vehicle to vehicle communication in Intelligent Transport System (ITS), WiMAX, WLAN, downlink of X-band and satellite communication and other applications operating in ultra wideband frequency range.

The new economy has seen tremendous strides since its early appearance at the onset of the industrial revolution in the 18th century. For decades, much of the items, including guns, tools, food, clothes, and homes, have been crafted or used from work by animals. This improved in the late 18th century with the introduction of the industrial methods. Industry 1.0's development was then a quick uphill climb leading up to the next manufacturing age – fourth Era. The summary of this evolution will be discussed here. This article takes a theoretical approach to looking at Business 4.0 as the Fourth generation. The study identifies three key elements of each transformation to deepen understanding of the phenomenon: technological, economic and demographic changes. In Business 4.0, Public Use Technology (PUT), Extreme competition and ageing demographics will allow the expansion quicker and broader. Although advances in Business 4.0 are more evaluative than transformative, their mixture and the context in which they develop forecast significant economic and social impacts that will in turn constitute a revolution.

Indian citrus ringspot virus (ICRSV) is a devastating pathogen that has a particularly deleterious effect on the ‘Kinnow mandarin’, a commercial citrus crop cultivated in the northwest of India. ICRSV belongs to the Mandarivirus genus within the family of Alphaflexiviridae and has a positive sense single-stranded RNA (ssRNA) genome consisting of six open reading frames (ORFs). Severe cases of ICRSV result in a significant reduction in both the yield and quality of crops. Consequently, there is an urgent need to develop methods to detect ICRSV in an accurate and timely manner. Current methods involve a two-step reverse transcription polymerase chain reaction (RT-PCR) that is time consuming. Here, we describe a novel, one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for the sensitive and rapid detection of ICRSV. To standardize the RT-LAMP assay, four different primers were designed and tested to target the coat protein gene of ICRSV. Amplification results were visualized by a color change after addition of SYBR Green I. The standardized RT-LAMP assay was highly specific and successfully detected all 35 ICRSV isolates tested from the Punjab and Haryana states of India. Furthermore, there was no cross-reaction with 17 isolates of five other citrus pathogens that are common in India. The ICRSV RT-LAMP assay developed in the present study is a simple, rapid, sensitive, specific technique. Moreover, the assay consists of only a single step and is more cost effective than existing methods. This is the first application of RT-LAMP for the detection of ICRSV. Our RT-LAMP assay is a powerful tool for the detection of ICRSV and will be particularly useful for large-scale indexing of field samples in diagnostic laboratories, in nurseries, and for quarantine applications.

Citrus tristeza virus (CTV) is the etiologic agent of the destructive Tristeza disease, a massive impediment for the healthy citrus industry worldwide. Routine indexing of CTV is an essential component for disease surveys and citrus budwood certification for production of disease-free planting material. Therefore, the present study was carried out to develop an efficient serological assay for CTV detection based on the RNA binding protein (CTV-p23), which is translated from a subgenomic RNA (sgRNA) that accumulates at higher levels in CTV-infected plants. CTV-p23 gene was amplified, cloned and polyclonal antibodies were raised against recombinant CTV-p23 protein. The efficacy of the produced polyclonal antibodies was tested by Western blots and ELISA to develop a quick, sensitive and economically affordable CTV detection tool and was used for indexing of large number of plant samples. The evaluation results indicated that the developed CTV-p23 antibodies had an excellent diagnostic agreement with RT-PCR and would be effective for the detection of CTV in field samples. Furthermore, CTV-p23 gene specific primers designed in the present study were found 1000 times more sensitive than the reported coat protein (CTV-p25) gene specific primers for routine CTV diagnosis. In silico characterizations of CTV-p23 protein revealed the presence of key conserved amino acid residues that involved in the regulation of protein stability, suppressor activity and protein expression levels. This would provide precious ground information towards understanding the viral pathogenecity and protein level accumulation for early diagnosis of virus.

Retinoblastoma (RB) is the most common paediatric intraocular tumour. Currently, chemotherapy is widely used to reduce the chance of metastasis as well as for vision salvage. The limitations of chemotherapy for RB include chemoresistance and cytotoxicity. Recently, immunotherapy is considered for treating chemoresistant cancers. Although, several molecular targets are available for immunotherapy in different cancers, we were interested in B7H3, as it was differentially expressed between retinoblastoma and retina in our earlier proteomics study. Hence, in this study we validated the previous finding by Western blotting and immunohistochemistry on primary RB tumor samples. The results suggest significantly increased expression of B7H3 in RB tumor samples compared to retina by western blotting. Immunohistochemistry revealed spatial, inter and intratumoral heterogeneity in the primary RB tumor sections. Correlation of the B7H3 expression with clinical and histopathological data revealed significantly increased expression of B7H3 in poorly differentiated, non-neural invasive tumors and lower expression in neural invasion and severe anaplastic areas of the tumors. B7H3 expression did not significantly vary between low-risk and high-risk tumors. The study also revealed considerably reduced infiltration of T lymphocytes in RB. We conclude that B7H3 is prominently expressed in primary RB tumors and could be used for targeted therapy.

Mycobacterium sp. is exhibiting complex evolution of antimicrobial resistance (AMR) and can therefore be considered as a serious human pathogen. Many strategies were employed earlier to evade the pathogenesis but AMR became threatened. Molecular tools employing bacteriophage can be an alternative to effective treatment against Mycobacterium. Phage treatment using phage-encoded products, such as lysins, causes lysis of cells; particularly bacteria could be used instead of direct use of these bacteriophages. Modern technologies along with bacteriophage strategies such as in silico immunoinformatics approach, machine learning, and artificial intelligence have been described thoroughly to escape the pathogenesis. Therefore, understanding the molecular mechanisms could be a possible alternative to evade the pathogenesis.

Synthetic dyes are a complex of aromatic molecules that possess coloration and are employed in many industries such as textile, leather, paper and pulp, plastics, food, pharmaceutical, cosmetics, etc. Based on the structural variation, different types of dyes such as azo, anthraquinone, reactive, triphenylmethane, acidic, basic, neutral, disperse, direct dyes, etc., are available and used in various industries. These dyes are released into the environment as effluents without proper treatment. These dyes are toxic, carcinogenic, and mutagenic, which affects life. They are resistant to conventional wastewater methods such as precipitation, filtration, and absorption. In the current scenario, enzymatic treatment is a most favorable approach to process these dyes because of its low energy cost and more ecofriendly nature. Peroxidases (EC1.11.1.x) are the heme-containing oxidoreductase enzymes that catalyze the H2O2-dependent oxidation of a variety of substrates such as dyes, small organic compounds, inorganic compounds, etc. Heme-containing peroxidases were initially classified into plant peroxidases and animal peroxidases. Subsequently, they were classified into various superfamilies on the basis of phylogenetic analysis: peroxidase-catalase, peroxidase-chlorite dismutase, peroxidase cyclooxygenase, di-heme peroxidase, and haloperoxidase. A new category of peroxidase was recently proposed, known as the family of dye-decolorizing (DyP-type) peroxidase. DyPs are the novel heme proteins that are structurally unrelated to other classical peroxidases. According to the PeroxiBase database, DyPs are categorized into four classes: A, B, C, and D. DyPs are receiving significant attention due to their stability in acidic conditions. Biochemical and mutational studies of DyPs have contributed to understanding the catalytic mechanism and role of amino acids in the enzyme catalysis. Furthermore, the structural and biophysical characterization of DyPs and comparative studies unveiled the substrate-binding sites, including the δ-site, the γ-site, and the surface-exposed redox-active residues; these sites are reported in other heme peroxidases also. DyPs have shown potential applications for the degradation of a wide range of dyes as well as lignin and β-carotene degradation. Therefore, DyPs are being used as a biocatalyst in industrial biotechnology. Further, protein engineering studies can be applied to increase the substrate-binding site and to enhance the degradation potential toward different substrates.

Mycobacterium sp. is exhibiting complex evolution of antimicrobial resistance (AMR) and can therefore be considered as a serious human pathogen. Many strategies were employed earlier to evade the pathogenesis but AMR became threatened. Molecular tools employing bacteriophage can be an alternative to effective treatment against Mycobacterium. Phage treatment using phage-encoded products, such as lysins, causes lysis of cells; particularly bacteria could be used instead of direct use of these bacteriophages. Modern technologies along with bacteriophage strategies such as in silico immunoinformatics approach, machine learning, and artificial intelligence have been described thoroughly to escape the pathogenesis. Therefore, understanding the molecular mechanisms could be a possible alternative to evade the pathogenesis.

Citrus yellow mosaic badnavirus (CMBV) is the etiologic agent of citrus yellow mosaic disease, which has caused serious economic losses to Indian citrus industry. CMBV is a quarantined pathogen that is geographically restricted to India. To prevent unintentional movement of the virus to other major citrus-growing countries in fruits, root stocks or grafted citrus plants and facilitate trade, a sensitive, validated diagnostic tool is needed. In the present study, we developed a SYBR Green real-time PCR-based method to detect and quantify CMBV in different tissues of infected Mosambi sweet orange (Citrus sinensis) and compared its sensitivity to conventional PCR protocols. Primers were designed to recognize a portion of the CMBV capsid protein gene. Conventional and real-time PCR were performed on several different tissues: shoot tips, leaves displaying typical CMBV symptoms, asymptomatic leaves, senescent leaves, thorns, green stems and feeder roots. The detection limit of CMBV by conventional PCR was 2.5 × 104 copies per 5 ng of total genomic DNA, while the detection limit of real-time PCR was found to be 4.6 × 102 virus copies per 5 ng of viral DNA. The viral load varied between different tissues. The highest concentration occurred in feeder roots (3.5 × 108 copies per 5 ng of total genomic DNA) and the lowest in thorns (1 × 106 copies per 5 ng of total genomic DNA). The variation in viral load within different tissues suggests movement of the virus within an infected plant that follows the path of photo-assimilates via the phloem. In symptomatic leaves, the CMBV concentration was highest in the lamella followed by midrib and petiole, suggesting that virus resides inside these sections of a leaf and side by side symptoms develop. On the other hand, in asymptomatic leaves, the petiole contained higher virus load than the lamella and midrib suggesting that the pathogen gets established from the stem through the phloem into petiole then infects the lamella and midrib. In addition to information on virus movement, the distribution of CMBV in different tissues helps with the selection of tissues with relatively higher viral load to sample for early and sensitive diagnosis of the disease, which will be useful for better management of the disease in endemic areas.

Candidatus Liberibacter asiaticus (CLas), a phloem-limited unculturable Gram-negative bacterium, causes citrus greening disease. The proteome analysis of CLas showed the presence of a heavy metal permease and Co/Zn/Cd cation exporter system. However, there is no designated metal uptake protein specific for the heavy metal permease in CLas. One of the metal uptake proteins, designated as CLas-ZnuA2, in our previous studies, showed a lower metal-binding affinity for Mn2+ and Zn2+ and was postulated to bind and transport metals rather non-specifically. The present study focused on the characterization of the heavy metal binding properties of CLas-ZnuA2 using SPR, CD, DSC and crystallographic studies. The crystal structure analysis of Cd2+ bound CLas-ZnuA2 showed octahedral geometry for Cd2+ binding as compared to a non-preferred square-pyramidal geometry for Mn2+ and Zn2+ binding in earlier reported crystal structures. In SPR analysis, the binding affinities of 4.7 × 10−6 M, 7.2 × 10−6 M, 5.3 × 10−5 M and 4.3 × 10−5 M for Hg2+, Cd2+, Ba2+ and Co2+ respectively were higher as compared to earlier reported values for Mn2+ and Zn2+. Likewise, CD and DSC analysis showed relatively higher thermal stability for CLas-ZnuA2 on heavy metal binding. Taken together with the expression of the permease and exporter system for heavy metals, our results indicate that CLas-ZnuA2 may be involved in sequestering and transport of various transition divalent metals in environmentally stressed conditions.

A miniaturized wideband Vivaldi antenna is presented in this paper. The proposed antenna is compact in size, has wide bandwidth and high radiation efficiency. The proposed antenna is simulated and fabricated on FR-4 substrate with overall dimensions of 20 × 17 mm2. The resonating frequency of the reported design is observed as 5.8GHz and impedance bandwidth is 2.67GHz (4.94GHz - 7.61GHz). The simulated radiation efficiency and gain are approximately 98.9% and 3.66dBi, respectively. The proposed design finds its applications in IoT, 5G, Wireless LAN (WLAN), Intelligent Transportation System (ITS) and Radio Frequency IDentification (RFID).

Bio composites offer a significant market in automotive and decking market but application in other sectors has been limited. An attempt has been made for the preparation of bio-degradable, eco-friendly green composites using the Coconut fiber and wheat husk weight with corn cob in the varying ratio. The various blends were molded and subjected to heating at 40°C for 48 hrs, Composite of coconut husk and wheat husk in the ratio of 15:85 gave the best tensile strength, this can be attributed to fibrous nature of coconut husk which has influence on tensile strength. The intertwining of the coconut husk initiates the crack propagation so the material shows the UTS of 7N/mm2 with a strain of 26% but with a stair-stepping stress-strain curve. Addition of corn makes this composite material into a hybrid composite. The proposed green composites are promising because they are renewable, biodegradable, and are better alternatives for non renewable composites.

Catabolite repressor activator (Cra) is a member of the LacI family transcriptional regulator distributed across a wide range of bacteria and regulates the carbon metabolism and virulence gene expression. In numerous studies to crystallize the apo form of the LacI family transcription factor, the N-terminal domain (NTD), which functions as a DNA-binding domain, has been enigmatically missing from the final resolved structures. It was speculated that the NTD is disordered or unstable and gets cleaved during crystallization. Here, we have determined the crystal structure of Cra from Escherichia coli (EcCra). The structure revealed a well-defined electron density for the C-terminal domain (CTD). However, electron density was missing for the first 56 amino acids (NTD). Our data reveal for the first time that EcCra undergoes a spontaneous cleavage at the conserved Asn 50 (N50) site, which separates the N-terminal DNA binding domain from the C-terminal effector molecule binding domain. With the site-directed mutagenesis, we confirm the involvement of residue N50 in the spontaneous cleavage phenomenon. Furthermore, the Isothermal titration calorimetry (ITC) assay of the EcCra-NTD with DNA showed EcCra-NTD is in a functional conformation state and retains its DNA binding activity.

A β-glucosidase gene from Putranjiva roxburghii (PRGH1) was heterologously expressed in Saccharomyces cerevisiae to enable growth on cellobiose. The recombinant enzyme was secreted to the culture medium, purified and biochemically characterized. The enzyme is a glycoprotein with a molecular weight of ∼68 kDa and exhibited enzymatic activity with β‐linked aryl substrates like pNP-Fuc, pNP-Glc, pNP-Gal and pNP-Cel with catalytic efficiency in that order. Significant enzyme activity was observed for cellobiose, however the enzyme activity was decreased with increase in chain length of glycan substrates. Using cellobiose as substrate, the enzyme showed optimal activity at pH 5.0 and 65 °C. The enzyme was thermostable up to 75 °C for 60 min. The enzyme showed significant resistance towards both glucose and ethanol induced inhibition. The recombinant S. cerevisiae strain showed advantages in cell growth, glucose and bio-ethanol production over the native strain with cellobiose as sole carbon source. In simultaneous saccharification and fermentation (SSF) experiments, the recombinant strain was used for bio-ethanol production from two different cellulosic biomass sources. At the end of the SSF, we obtained 9.47 g L−1 and 14.32 g L−1 of bio-ethanol by using carboxymethyl cellulose and pre-treated rice straw respectively. This is first report where a β-glucosidase gene from plant origin has been expressed in S. cerevisiae and used in SSF.

The 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) transformylase/inosine monophosphate (IMP) cyclohydrolase (ATIC) catalyzes final two steps of purine nucleotide de novo biosynthetic pathway. This study reports the characterization of ATIC from Staphylococcus lugdunensis (SlugATIC). Apart from kinetic analysis and a detailed biophysical characterization of SlugATIC, the role of ATIC in cell proliferation has been demonstrated for the first time. The purified recombinant SlugATIC and its truncated domains exist mainly in dimeric form was revealed in gel-filtration and glutaraldehyde cross-linking studies. The two activities reside on separate domains was demonstrated in kinetic analysis of SlugATIC and reconstituted truncated N-terminal IMP cyclohydrolase (IMPCHase) and C-terminal AICAR transformylase (AICAR TFase) domains. Site-directed mutagenesis showed that Lys255 and His256 are the key catalytic residues, while Asn415 substantially contributes to AICAR TFase activity in SlugATIC. The differential scanning calorimetry (DSC) analysis revealed a molten globule-like structure for independent N-terminal domain as compared with a relatively stable conformational state in full-length SlugATIC signifying the importance of covalently linked domains. Unlike reported crystal structures, the DSC studies revealed significant conformational changes on binding of leading ligand to AICAR TFase domain in SlugATIC. The cell proliferation activity of SlugATIC was observed where it promoted proliferation and viability of NIH 3T3 and RIN-5F cells, exhibited in vitro wound healing in NIH 3T3 fibroblast cells, and rescued RIN-5F cells from the cytotoxic effects of palmitic acid and high glucose. The results suggest that ATIC, an important drug target, can also be exploited for its cell proliferative properties.

The biosorption ability of ‘Moringa oleifera ‘seed powder for the expulsion of cadmium from wastewater was investigated by performing packed column tests. Column tests were conducted as a function of biosorbent dose, influent flow rate and feed sorbate concentration. The findings from column operations revealed that values of maximum cadmium uptake, exhaustion and breakthrough time enhances with rise in bed depth, reduces with rise in feed volumetric flowrate and initial cadmium concentration. At 6 g of Moringa oleifera dose, 2 mL/min of flow rate and 300 µg/L of influent cadmium, the cadmium uptake capacity was found to be 65.44, µg/g. The continuous packed column data for cadmium were well fitted to the Thomas model with the regression coefficient ranges 0.966–0.993. Moreover, the qth values obtained from Thomas model are similar to the experimental breakthrough capacities qexp. The FTIR results reveals that the Moringa oleifera seed contains various active functional groups which are appears to be played a significant role in biosorption of cadmium ions. From practical view, the Moringa oleifera seed could be utilized as safe, inexpensive, sustainable and eco-friendly biosorbent for the elimination of cadmium from drinking water using continuous column mode and also this technology could be considered as an ideal for household usage in rural communities.

The native Putranjiva roxburghii family 1 glycoside hydrolase enzyme showed β-D-fucosidase activity in addition to β-D-glucosidase and β-D-galactosidase activities reported in our previous study. A single step concanvalin A affinity chromatography for native PRGH1 improved the yield and reduced the purification time. The PRGH1 gene was cloned and overexpressed in E. coli . The full length gene contained an ORF of 1617 bp encoding a polypeptide of 538 amino acids. The amino acid sequence of PRGH1 showed maximum similarities to β-glucosidases and myrosinases. Both native and recombinant protein showed maximum hydrolytic activity for p NP-Fuc followed by p NP-Glc and p NP-Gal. Significant enzyme activity was also observed for cellobiose, however it decreased with increase in chain-length for glycan substrates. The enzyme showed significant resistant to D-glucose concentration up to 500 mM. Mutational studies confirmed the predicted catalytic acid/base Glu173 and nucleophile Glu389 as key residues for its activity. Moreover, Glu446 and Asn172 played essential role in substrate binding by interacting with the −1 subsite of substrates. Bioinformatic analysis suggested the possible reasons for the broad substrate specificity and other properties of the enzyme. PRGH1 had high sequence similarity towards S-glucosidase and may be involved in defence. The broad specificity, catalytic efficiency and thermostability make PRGH1 potentially an important industrial enzyme.

The attainment of new function by a protein is achieved through convergent/divergent evolution. In present work, the sequence analysis of a 34 kDa protein from Putranjiva roxburghii, earlier reported as a potent trypsin inhibitor, showed resemblance to some of the wound inducible and vegetative storage proteins. A detailed sequence analysis revealed that these proteins belong to PNP-UDP family. In case of P. roxburghii protein, an approximately 46 residue insert disrupts the PNP domain. Similar disruption of PNP domain is observed in related plant proteins. The characterization of recombinant full length and truncated (without 46 residue insert) forms of P. roxburghii PNP family protein (PRpnp) unraveled that trypsin inhibitory active site is located within the insert. The truncated form containing uninterrupted PNP domain showed strong PNP enzymatic activity where it hydrolyzed the N-glycosidic bond of inosine and guanosine. The full length protein, however, showed weak PNP enzyme activity which may be due to presence of the insert. These results indicate towards the neofunctionalization of PRpnp to a potent trypsin inhibitor through an insert containing inhibitory residue to cater to the needs of plant defense. The similar wound inducible and vegetative storage proteins may have also evolved due to evolutionary needs.

Magnetic-field (H-Field) forming using planar multicoil transmitter antenna is presented. To address angular misalignment in inductive coupling systems, three orthogonal H-field components are formed in near-field zone of the transmitter antenna. This proposed magnetic field forming is performed by analyzing near-field distributions of various combinations of spatially distributed coils. The analytical study evolves into the planar multicoil design achieving three orthogonal components of H-field formed in target receiver area. The proposed antenna is realized in printed circuit board technology, simulated, and measured to validate the claim. The results corroborated with the analytical results and proved potential of the proposed design suitable for inductive coupling based applications like radio frequency identification, wireless charging, near-field communication, and medical implantation.

Globulins are a major class of seed storage proteins which were thought to be enzymatically inactive. These proteins belong to the most ancient cupin superfamily. They can be graded into 11S legumin type and 7S vicilin type based on their sedimentation coefficients. Members from both classes share structural homology are thought to have evolved from either one-domain germin predecessor by duplication or by horizontal gene transfer of two-domain gene from bacteria to eukaryotes. Globulins are known to define the nutritional quality of the seeds, however, they are also involved in sucrose binding, desiccation, defense against microbes, hormone binding and oxidative stress etc. Major drawback with globulins is their tendency to bind to IgE. Studying structural-functional behavior of such protein can help in modifying proteins for enhanced functionality in food processing industries.

A multipurpose near- and far-field switched multiband reader antenna for radio frequency identification (RFID) applications is presented. The proposed antenna is simple, planar, and has three modes of operating frequencies: 915 MHz, 2.45 GHz, and 5.8 GHz. The prototype is realized using nonuniformly distributed turns coil for robust reactive and radiated field performance, leading to a multipurpose near- and far-field antenna in a single unit. Results show performance of the proposed multipurpose antenna in both near-field and far-field region and demonstrate its suitability for various RFID applications.

A periplasmic solute binding protein from second of the two gene clusters of Znu system in CLA (CLas-ZnuA2) belong to Cluster A1 family of solute binding proteins (SBPs). The crystal structures in metal-free, intermediate and metal-bound states, in the previous study, revealed the unusual mechanism of metal binding and release for CLas-ZnuA2. Although CLas-ZnuA2 showed maximum sequence identity to the Mn/Fe-specific SBPs, the mechanistic resemblance seems to be closer to Zn-specific SBPs of Cluster A-I family. The present study reports the binding affinity studies using SPR and CD and crystal structure of CLas-ZnuA2 in Zn2+-bound state. Despite a similar overall structure, there are noticeable differences at the metal binding site. The SPR and CD analysis confirmed our previous observation that CLas-ZnuA2 exhibits a low metal-binding affinity. The low metal-binding affinity of CLas-ZnuA2 could be attributed to the presence of a proline in linker helix resulting in relatively higher bending and rigidity of the same. This structural feature fixes the C-domain similar to metal-bound states of related SBPs. Further, the binding of both Mn2+ and Zn2+ occurs pentavalently with square pyramidal geometry not preferred by either. The site-specific positive Darwinian selection analysis showed that the proline in linker helix is under purifying selection and might have diverged long ago. Our structural and evolutionary analyses suggest that CLasZnua2 might have evolved, particularly for plant pathogens, to facilitate transport of both Mn2+ and Zn2+, with reversible binding to Zn2+, unlike other Mn-binding SBPs (PsaA).

Due to the advancements in modern technology, there is an immense use of Internet worldwide. For the smooth transmission of the information from one end to other, systems having characteristics of more bandwidth and high capacity are required. Hence, optical fiber is used for this purpose. Like all other transmission media, optical fiber also suffers from signal deteriorating factor that includes attenuation, dispersion, and other nonlinear effects. Hence, there is need for their compensation that includes utilizing dispersion compensation techniques like DCF, FBG, EDC, and digital filters. The work in this paper evaluates the performance of distinct apodized profiles for FBG. Basically, three types of profiles: Gaussian, uniform, and hyperbolic tangent are executed and investigated at a data rate of 100 Gbps over a transmission distance of 120 km. After analyzing the outcomes, these techniques are then compared in order to get the best apodization scheme for FBG to implement so as to achieve less dispersion at the receiver end.

Because of reactive power requested by the industrialists and agriculturists, three-phase induction motors composed at evaluated voltage are working under the appraised voltage particularly in the rural areas, a long way from the utility focuses. The target of this research article is to enhance torque, effectiveness, power developed, and power factor by using double-cage winding with a new combination of stator and rotor design. The rim_003 rotor bar is used with so_012 parallel tooth opening flat bottom stator with different arrangements of stator winding, which are used for 5 HP three-phase squirrel cage induction motor. JMAG Express is used to figure the execution enhancement of motor under appraised voltage.

Molecular recognition between nucleic acids has proven to be a powerful tool for designing hybridization probes for the detection of DNA and RNA sequences. Most detection probes rely on the conjugation of small molecule dyes to nucleic acids for fluorescence output, which is not cost-effective and also limits their applications in vivo, as they are not genetically encodable. More affordable sensors devoid of any chemical labeling are needed that show high fluorescence output and are genetically encodable. Here, we have designed a label-free Baby Spinach-based minimal modified sensor (BSMS) for the analysis of nucleic acids. The minimal modification in the sensor design reduces the complexity of the design, and provides additional stabilization after binding the target nucleic acids, leading to a high fluorescence output. BSMS is able to detect both DNA and RNA of potentially any lengths and is based on a Baby Spinach aptamer that binds and enhances the fluorescence of a small molecule dye. BSMS shows specificity towards its analyte in the presence of other sequences and selectively differentiates between closely related sequences. BSMS comprises genetically encodable unmodified RNA and has been shown to function at ambient temperature, and thus is anticipated to provide nucleic acid monitoring in vivo.

Earlier reported crystal structure of CLasTcyA revealed unique features like relatively a larger substrate binding pocket, an extended C-terminal loop restricted by a disulfide bond and involvement of residues from hinge region in substrate binding. In present study, CLasTcyA mutants were created to evaluate the importance of these unique features through biophysical characterization. The Val58 in CLasTcyA was replaced by Trp, conserved in most cystine binding proteins, to reduce the size of the binding pocket. All other mutations were created in CLasTcyAV58W mutant as the presence of Trp could be used for intrinsic fluorescence studies. The CLasTcyAV58W showed a noticeable increase in binding affinity and thermal stability as compared to the native form. The mutation of two cysteines in triple mutant CLasTcyAV58W/C212S/C239S, removal of C-terminal extended loop in truncated CLasTcyAV58W/C212S and mutation of His95 from hinge region in the double mutant CLasTcyAV58W/H95A showed a marked decrease in stability-indicating the importance of the unique features in structure of CLasTcyA. The bioinformatics-based virtual screening was employed to screen the potential inhibitor molecules for detailed future studies. The results clearly establish the importance of unique features in structure-function relationship of CLasTcyA.

Background: Microtubules are dynamic protein filaments that are crucial for cell division and constitute key elements of the cytoskeleton. They are assembled from αβ-tubulin heterodimers that form hollow cylindrical structures. There is a large number of naturally occurring compounds that are known to interact with tubulin, including alkaloids, macrolides and peptides, which are collectively called microtubule-targeting agents (MTAs). Based on their activities, MTAs can be classified as microtubule-stabilizing agents (MSAs) that enhance MT assembly, and microtubule-destabilizing agents (MDAs) that suppress MT assembly. The chemical structure of these drugs and their binding mode to microtubules varies greatly amongst each other and confer the ability to act either synergistically or competitively on microtubules. Owing to their effects on microtubule dynamics, MTAs are of great interest and widely used in a variety of medical applications as antiparasitic agents, herbicides and, most importantly, as chemotherapeutic drugs used for the treatment of cancer. In the past years, we and others solved the structures of a large number of different MTAs bound to tubulin to high resolution using X-ray crystallography. Very recently, however, with the advent of the “Resolution Revolution” in cryo-electron microscopy (cryo-EM), atomic structures of known MSAs bound to microtubules have also been obtained. These cryo-EM structures confirmed that the sites and modes of binding described in the previous X-ray crystallographic studies are similar in the context of the assembled microtubule but additionally explain the effects of MTAs on lattice parameters in microtubules and the lateral contacts between protofilaments, especially at the microtubule seam. Methods: Cryo-EM and model building. Results: We solved several high-resolution structures of microtubules bound to novel MSAs and identified differential binding modes in comparison to that revealed by X-ray crystallography. Conclusion: Cryo-EM and X-ray crystallography can be used in a complementary manner to investigate the molecular mechanism of action of MSAs in detail.

SUMMARY Dendritic cell (DC) activation is a critical step for anti-tumor T cell responses. Certain chemotherapeutics can influence DC function. Here we demonstrate that chemotherapy capable of microtubule destabilization has direct effects on DC function; namely, it induces potent DC maturation and elicits anti-tumor immunity. Guanine nucleotide exchange factor-H1 (GEF-H1) is specifically released upon microtubule destabilization and is required for DC activation. In response to chemotherapy, GEF-H1 drives a distinct cell signaling program in DCs dominated by the c-Jun N-terminal kinase (JNK) pathway and AP-1/ATF transcriptional response for control of innate and adaptive immune responses. Microtubule destabilization, and subsequent GEF-H1 signaling, enhances cross-presentation of tumor antigens to CD8 T cells. In absence of GEF-H1, anti-tumor immunity is hampered. In cancer patients, high expression of the GEF-H1 immune gene signature is associated with prolonged survival. Our study identifies an alternate intracellular axis in DCs induced upon microtubule destabilization in which GEF-H1 promotes protective anti-tumor immunity., Graphical Abstract, In Brief Certain chemotherapeutics elicit potent anti-tumor immunity. Kashyap et al. demonstrate that microtubule-destabilizing chemotherapeutics induce maturation of dendritic cells through activation of microtubule-associated protein GEF-H1. This leads to effective priming of CD8 T cells against tumor antigens. GEF-H1 is critical for anti-tumor immunity of microtubule-targeting chemotherapy.

25 p.-5 fig.-1 tab. + 6 fig. supl.+ 1 tab. supl., αβ-Tubulin is a validated target for anticancer drug discovery, and molecules binding to this protein are used to treat several types of tumors. Here, we report on a combined X-ray crystallography and molecular dynamics approach to study drug binding within the colchicine site of αβ-tubulin, focusing on plinabulin, an agent currently in phase 3 clinical testing for the treatment of cancer and chemotherapy-induced neutropenia. We found that plinabulin is more persistently bound to the colchicine site of βII- compared to βIII-tubulin, allowing for a prediction of isotype-expression-dependent drug sensitivity. Additionally, computational residence time and exit paths from the βII-tubulin were compared between plinabulin and two other compounds, colchicine and combretastatin-A4. The former displayed the highest residence time, followed by plinabulin and then distantly by combretastatin-A4. Our combined experimental and computational protocol could help to investigate anti-tubulin drugs, improving our understanding of their mechanism of action, residence time, and tubulin isotype selectivity., This work was financially supported by BeyondSpring Pharmaceuticals Inc. (to M.O.S. and A.C.) and by grants from the Ministerio de Ciencia Innovacion y Universidades (BFU2016-75319-R; to J.F.D.), from the Swiss National Science Foundation (31003A_166608, to M.O.S.) and from the Regione Lombardia (Accordo per la Ricerca e l’Innovazione).

The dye-decolorizing peroxidases (DyPs) belong to a unique heme peroxidase family for their biotechnological potential to detoxify synthetic dyes. In this work, we have biochemically and structurally characterized the dye-decolorizing peroxidase from Bacillus subtilis (BsDyP). The biochemical studies of BsDyP demonstrate that pH 4.0 is optimum for the oxidation of malachite green (MG) and methyl violet (MV). However, it oxidizes the MG with higher catalytic efficiency (kcat/Km = 6.3 × 102 M−1s−1), than MV (kcat/Km = 5.0 × 102 M−1s−1). While reactive black 5 (RB5) is oxidized at pH 3.0 with the catalytic efficiency of kcat/Km = 3.6 × 102 M−1s−1. The calculated thermodynamic parameters by isothermal titration calorimetry (ITC) reveal the feasibility and spontaneity of dyes binding with BsDyP. Further, the crystal structures of a HEPES bound and unbound of BsDyP provide insight into the probable binding sites of the substrates. In BsDyP-HEPES bound structure, the HEPES-1 molecule is found in the heme cavity at the γ-edge, and another HEPES-2 molecule is bound ~16 A away from the heme that is fenced by Ile231, Arg234, Ser235, Asp239, Glu334, and surface-exposed Tyr335 residues. Furthermore, the molecular docking, simulation, and MMPBSA studies support the binding of dyes at both the sites of BsDyP and produce lower-energy stable BsDyP-dyes complexes. Here, the BsDyP study allows the identification of its two potential binding sites and shows the oxidation of a variety of dyes. Structural and functional insight of BsDyP will facilitate its engineering for the improved decolorization of dyes.

Doublecortin is a microtubule-associated protein produced during neurogenesis. The protein stabilizes microtubules and stimulates their polymerization, which allows migration of immature neurons to their designated location in the brain. Mutations in the gene that impair doublecortin function and cause severe brain formation disorders are located on a tandem repeat of two doublecortin domains. The molecular mechanism of action of doublecortin is only incompletely understood. Anti-doublecortin antibodies, such as the rabbit polyclonal Abcam 18732, are widely used as neurogenesis markers. Here, we report the generation and characterization of antibodies that bind to single doublecortin domains. The antibodies were used as tools to obtain structures of both domains. Four independent crystal structures of the N-terminal domain reveal several distinct open and closed conformations of the peptide linking N- and C-terminal domains, which can be related to doublecortin function. An NMR assignment and a crystal structure in complex with a camelid antibody fragment show that the doublecortin C-terminal domain adopts the same well defined ubiquitin-like fold as the N-terminal domain, despite its reported aggregation and molten globule-like properties. The antibodies' unique domain specificity also renders them ideal research tools to better understand the role of individual domains in doublecortin function. A single chain camelid antibody fragment specific for the C-terminal doublecortin domain affected microtubule binding, whereas a monoclonal mouse antibody specific for the N-terminal domain did not. Together with steric considerations, this suggests that the microtubule-interacting doublecortin domain observed in cryo-electron micrographs is the C-terminal domain rather than the N-terminal one.

By reflecting on running the open-access online “race” journal darkmatter, the author examines the crisis of academic journal publishing. This crisis is manifested by the transformations in the economics of journal production, especially with the rise of open-access publishing and the neoliberalization of the academy; the developments in digital communications; and the global consolidation of Western corporate publishing power in “info-capitalism.” The evolution of darkmatter is poised in terms of negotiating this crisis as opportunity and risk. The digital journal is able to produce a range of multimedia outputs addressing the interdisciplinary fields of race and postcolonial study. The challenge of emerging Web projects and the proliferation of online information potentially repositions darkmatter as the privileged site for the publication of scholarly research. The essay speculates on this critical journal in a time of “information overload” as a site of “autonomous resistance” in the increasing commodification of knowledge.