Efficient Protocol Research Articles

Optimization of an Efficient Micropropagation Protocol and the Assessment of Genetic Fidelity of Solanum tuberosum L. cv Kufri Neelkanth

The aim of our experiment was to determine the optimal types and concentrations of plant growth regulators used during different stages of micropropagation and assess the genetic fidelity of tissue culture raised plants of a purple colored cv Kufri Neelkanth by using reliable markers. According to the shoot bud initiation data, it was found that utilizing a combination of 0.25 mg l-1 BAP and 0.5 mg l-1 Kinetin resulted in the greatest number of shoots per explant (3.5±0.12) within a timeframe of (3.3±0.08) days. The maximum number of in vitro shoots per shootlet (10.4±0.59) were recorded when auxins were used in combination with NAA (0.01 mg l-1 NAA) and Kinetin (0.25 mg l-1) for shoot proliferation. In vitro root initiation was observed in (2.1±0.07) days on MS medium fortified with 2.5 mg l-1 NAA. The maximum number of in vitro roots per shoots (13.0±0.55) were observed when MS media fortified with 2.5 mg l-1 IBA. Maximum 100% rooting was observed in all MS media supplemented with different concentrations of auxins. In vitro raised plants were assessed for genetic fidelity by using twenty RAPD primers (genetic markers). Out of twenty primers used only four primers produced amplification. DNA banding patterns of all tissue culture raised plants and mother plants were monomorphic showing true to type planting material. This protocol for tissue culture propagation along with testing its genetic fidelity could be useful for better conservation of germplasm and genetic transformation studies in potato.

Clustering Algorithms and Optimization Strategies in WSNs: A State-of-the-Art Review

Wireless Sensor Networks (WSNs) have emerged as a groundbreaking technology for collecting data on various physical phenomena, offering the potential to revolutionize infrastructure systems. The paper provides an exhaustive exploration of the existing literature on clustering algorithms within WSNs and the diverse optimization techniques proposed for cluster head selection. A comprehensive analysis of various routing protocols, mobility-based solutions, and energy conservation strategies is presented, shedding light on their respective strengths and limitations. The examination encompasses a wide spectrum of approaches, including hierarchical cluster-based routing, mobile sensor integration, and energy-efficient techniques inspired by industry standards like ZigBee. The paper further explores ecological monitoring applications, redundancy reduction mechanisms, and power- efficient hierarchical routing protocols, offering valuable insights into their contributions to the field. This review paper aims to consolidate and synthesize the wealth of knowledge in the domain of clustering algorithms and optimization techniques for WSNs. By assessing the strengths and weaknesses of these methodologies, it seeks to provide a comprehensive resource for researchers and practitioners in the field, ultimately contributing to the advancement of energy-efficient and reliable wireless sensor networks. Keywords— APTEEN, EARP, ECCP, GPS, HEED, LEACH, PEGASIS, TEEN, WSN.

An α, β-unsaturated ketone alkylation and efficient reduction protocol for the synthesis of 3α-hydroxy-3β-methyl-4,4-dimethyl-5α-21-bromo-19-nor-pregnan-20-one

Alkyl-substituted testosterone derivatives are promising platforms for new drug discovery in medicinal chemistry. This approach provides a simple and efficient method for introducing an alkyl substituent to steroids at the C-4 position. In this study, the novel compound 3α-hydroxy-3β-methyl-4,4-dimethyl-5α-21-bromo-19-nor-pregnan-20-one is synthesized from 19-nor-testosterone. The protocol involves methylation of the dienolate, reduction of the alkene, the Grignard reaction of the carbonyl group, a Wittig reaction, hydroboration with BH3, the oxidation and bromination with a 49% overall yield. For the methylation and reduction steps, the effects of the base, solvent, and reactant ratio on the conversion and yield are investigated. The structures of the synthesized compounds are determined by nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS) (electrospray ionization (ESI)).

A Novel Access and Handover Authentication Scheme in UAV-Aided Satellite-Terrestrial Integration Networks Enabling 5G

As an outlook of the terrestrial network, the flexible Unmanned Aerial Vehicle (UAV)-aided satellite-terrestrial integration network would have numerous prospective applications such as service enhancement, maritime communication, military, and emergency communication, which is getting significant attention. However, the public and open-access network must result in various imperative risks including impersonation, sensitive data, and privacy disclosure. Due to several unique characteristics including long transmission distance, unstable communication environment, resource-limited and highly dynamic characteristics of UAVs, diversified terminals that lack the ability of accessing the satellite and may be resource-limited, there are new challenges for the access authentication process in the UAV-Aided Satellite-Terrestrial Integration Networks with 5G. In this paper, we present a novel Physically Unclonable Function (PUF)-based access authentication scheme consisting of two access authentication protocols for the UAV and the ground terminals like 5G User Equipments (UEs), respectively. Two access authentication protocols for the drone and the terminals can both achieve mutual authentication, key agreement, and privacy protection with distinct advantages of no need to store secret key and supporting physical attack resistance. Finally, we propose an efficient handover authentication protocol executed by the ground terminals when the UAV is required to switch. We employ different security analysis tools to analyze the security of all proposed protocols, as well as present informal security analysis on various security properties. The performance comparison and evaluations show our protocols have better advantages.

SecBerg: Secure and Practical Iceberg Queries in Cloud

Secure queries are fundamental to data security, particularly in cloud databases. In data analytics, one of the common and practical queries is the iceberg query that can find aggregate values above a specified threshold. However, existing secure aggregate query schemes: i) are unable to support secure iceberg queries equipped with the HAVING clause; ii) only consider additive aggregate functions; and iii) suffer from performance issues due to the use of homomorphic encryption to encrypt databases. In this paper, we present a secure iceberg query scheme, SecBerg, to support both addition-based and comparison-based aggregate functions and ensure high efficiency and security simultaneously. To make it possible, we propose a secure bitmap index system to encode database values and pioneer the use of the arithmetic secret sharing technique to protect databases in the cloud environment. Furthermore, we carefully design efficient and secure protocols over arithmetic secret sharing to construct our SecBerg. Extensive evaluations are conducted, and the results indicate that SecBerg is significantly more efficient than the state-of-the-art relevant scheme in computational overhead and can attain orders of magnitude performance improvement at best.

In vitro Regeneration of Multiple Shoots in Abelmoschus esculentus (L.) Moench (Okra) via Apical Shoot Meristem Culture

Introduction: To develop the efficient protocol for multiple shoot regeneration of A.esculentus by apical root culture method. Abelmoschus esculentus L., commonly known as okra, is a member of the Malvaceae family, which is widely consumed due to its high nutrient content and potential therapeutic properties. Okra contains various bioactive compounds that ex-hibit antibacterial properties and may be useful in treating type-2 diabetes, digestive diseases, and liver detoxification. To select the plant for the present and prepare the efficient protocol for the development of multiple shoot regeneration culture method. Methods: In this study, we developed an efficient protocol for multiple shoot regeneration of A. esculentus using the apical shoot culture method. Mature shoot apex explants of the germinated A. esculentus genotype CoBhH1 were cultured on Murashige and Skoog (MS) medium supple-mented with different concentrations of cytokinins (BAP and TDZ) and auxins (IAA and NAA) to determine the optimal conditions for shoot induction. Results: The highest number of multiple shoots (27.04 shoots) was obtained with 0.8 mg/L TDZ. Excised shoots were cultured on MS medium supplemented with GA3, which induced elongation of the shoots to a maximum of 8-10 cm. Regenerated plantlets were successfully transferred to soil, with a 100% survival rate and no differences in morphology or growth characteristics com-pared to control plants. Rooting was achieved with 1 mg/L IBA. Conclusion: This study provides an efficient protocol for multiple shoot regeneration of A. esculentus through apical shoot culture, which has potential applications in plant breeding and genetic engineering.

In vitro regeneration, phytochemical profiling and antioxidant activity in Ruta chalepensis plants established from alginate encapsulated synthetic seeds

We developed a highly efficient protocol for the short-term conservation and in vitro regeneration of Ruta chalepensis L. This involved encapsulating nodal segments (NS) in a sodium-alginate solution (3% w/v) and calcium chloride (100 mM) to produce synthetic seeds. The effect of various concentrations of plant growth regulators (PGRs) on the conversion of synthetic seeds into plantlets was examined and the best results, with a conversion rate of 96.8%, were obtained on Murashige and Skoog (MS) medium supplemented with 5.0 μM 6-benzyladenine (BA) and 0.5 μM α-naphthalene acetic acid (NAA) after 12 weeks of culture. When stored at 4 °C for different durations (2-12 weeks), encapsulated and non-encapsulated NS displayed distinct conversion responses. Encapsulated NS stored at 4 °C for 10 weeks exhibited a regeneration rate of 72.7%, while non-encapsulated NS did not survive under the same conditions. The proliferated shootlets derived from cold-stored NS were successfully rooted in agar-solidified MS medium containing 0.5 μM indole-3-butyric acid (IBA) and successfully acclimatized to ex vitro conditions. UV-Visible spectroscopy (UV), gas chromatography-mass spectrometry (GC–MS), and high-performance liquid chromatography (HPLC) were used to identify bioactive chemicals in the aqueous methanolic extracts of the acclimatized plants and compared with the donor plant. The bioactive chemicals were found considerably higher in the acclimatized plants (Phenols 95.0 mg GAE/g DW, flavonoids 99.2 mg QE/g DW, tannins 38.7 mg TAE/g DW, hesperidin 94.5 µg/mg, and quercetin 25.4 µg/mg). The antioxidant potential was also evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and demonstrated high scavenging activity at a concentration of 800 µg/mL in the acclimatized plants (83.0%). This study represents the first successful demonstration of in vitro regeneration, phytochemical profile analysis, and assessment of antioxidant activity in R. chalepensis plants established from synthetic seeds following short-term cold storage.

Energy Efficient Three-Factor Authentication in Wireless Sensor Networks With Resisting Insider Attacks

Wireless Sensor Networks (WSNs) play an important role in many fields such as industry, medical treatment, and national defense. Due to the limited node resources, there has always been an intractable contradiction between WSNs security and energy consumption. This contradiction needs urgent attention, especially with the increasing capability of attackers. User authentication is a key technology to ensure the security of WSNs. Unfortunately, recent insider attacks have shown vulnerabilities in the registration and authentication processes of traditional protocols. This paper analyzes the common types of insider attacks and focuses on explaining the logic of the offline guess attack launched by insiders. In order to uphold the concept of green communication, this paper proposes a new energy efficient protocol based on the widely used three-factor authentication. For the first time, the protocol combines lightweight data hiding technology to protect against offline guess and insider attacks. Experiments and analyses show that the proposed scheme not only solves the insider attack problem, but also reduces the energy consumption compared with the current typical schemes.

Protein Loop Modeling Using AlphaFold2.

The functions of proteins are largely determined by their three-dimensional (3D) structures. Loop modeling tries to predict the conformation of a relatively short stretch of protein backbone and sidechain. It is a difficult problem due to conformational variability. Recently, AlphaFold2 has achieved outstanding results in 3-D protein structure prediction and is expected to perform well on loop modeling. In this paper, we investigate the performances of AlphaFold2 variants on popular loop modeling benchmark datasets and propose an efficient protocol of using AlphaFold2 for loop modeling, called IAFLoop. To predict the structure of a loop region, IAFLoop gives a moderately extended segment of the target loop region as input to AlphaFold2, runs a fast version of AlphaFold2 using a reduced database without ensembling, and uses RMSD based consensus scores to select the final output models. Our experimental results on benchmark datasets show that IAFLoop generated highly accurate loop models. It achieves comparable performance to the original application of AlphaFold2 in terms of RMSD error, and achieving much better results on some targets, while only using half of the time. Compared to the best previous methods, IAFLoop reduces the RMSD error by almost half on the 8-residual loop dataset, and more than 70% on the 12-residual loop dataset.

International biathlon season during the COVID-19 pandemic was based on frequent on-site PCR-testing protocol with rapid result management.

The resumption of professional sports during the COVID-19 pandemic has been described in team sports but less in individual sports. The International Biathlon Union implemented a COVID-19 concept for the 2020-2021 season aimed to mitigate the risks of transmission by rules designated for the professional biathlon environment. The "bubble" model was based on regular reverse transcript polymerase chain reaction (PCR) testing with rapid results and efficient result management protocol. The objective of this study was report incidence and transmission of SARS-CoV-2 among professional biathletes and staff undergoing frequent PCR testing and risk reduction measures during the international season 2020-2021. The efficiency of risk mitigation measures was also evaluated based on the incidence data. During the 4-month season, altogether 22,182 SARS-CoV-2 PCR tests were conducted on all individuals participating in international biathlon season (athletes, team staff and organizing committee). Ninety-six (0.4%) PCR tests were positive and 30% of the positive PCR tests were considered "persistent positive" following recovery from a recent COVID-19 infection. No transmission events were detected following contact with "persistent positive" cases during the season. A great majority of the positive PCR tests were recorded during the first days after arrival in the "bubble", often in the first entry test taken by the on-site laboratory. In conclusion, a "bubble model" based on frequent PCR testing and hygiene rules was efficient in keeping the infection rate low. The competition activity including international travel was safe, and most of the infections seemed to originate from outside of the "bubble".

Metaheuristic Based Energy Efficient Routing Protocol in MANET Using Battle Royale Optimization

Metaheuristic Based Energy Efficient Routing Protocol in MANET Using Battle Royale Optimization

Efficient Protocol for Computing MCD Spectra in a Broad Frequency Range Combining Resonant and Damped CC2 Quadratic Response Theory.

Coupled cluster response theory offers a path to high-accuracy calculations of spectroscopic properties, such as magnetic circular dichroism (MCD). However, divergence or slow convergence issues are often encountered for electronic transitions in high-energy regions with a high density of states. This is here addressed for MCD by an implementation of damped quadratic response theory for resolution-of-identity coupled cluster singles-and-approximate-doubles (RI-CC2), along with an implementation of the MCD term from resonant response theory. Combined, damped and resonant response theory calculations provide an efficient strategy to obtain MCD spectra over a broad frequency range and for systems that include highly symmetric molecules with degenerate excited states. The protocol is illustrated by application to zinc tetrabenzoporphyrin in the energy region of 2-8 eV and comparison to experimental data. Timings are reported for the resonant and damped approaches, showing that a greater part of the calculation time is consumed by the construction of the building blocks for the final MCD ellipticity. A recommendation on how to use the procedure is outlined.

Functionalization of octaspherosilicate (HSiMe2O)8Si8O12 with buta-1,3-diynes by hydrosilylation

Hydrosilylation with octaspherosilicate (HSiMe2O)8Si8O12 (1) has provided hundreds of molecular and macromolecular systems so far, making this method the most popular in the synthesis of siloxane-based, nanometric, cubic, and reactive building blocks. However, there are no reports on its selective reaction with 1,3-diynes, which allows for the formation of new products with unique properties. Therefore, herein we present an efficient protocol for monohydrosilylation of symmetrically and non-symmetrically 1,4-disubstituted buta-1,3-diynes with 1. The compounds obtained bear double and triple bonds and other functionalities (e.g., Br, F, OH, SiR3), making them highly desirable, giant building blocks in organic synthesis and material chemistry. These compounds were fully characterized by 1H, 13C, 29Si, 1D NOE, 1H–13C HSQC NMR, FT–IR, and MALDI TOF MS, EA, UV–Vis, and TGA analysis. The TGA proved their high thermal stability up to 427 ℃ (Td10%) for compound 3j.

A blockchain-enabled privacy-preserving authentication management protocol for Internet of Medical Things

Over the last decade, with the increasing popularity and usage of the internet of things worldwide, Internet of Medical Things (IoMT) has emerged as a key technology of the modern era. IoMT uses Artificial Intelligence, 5G, big data, edge computing, and blockchain to provide users with electronic medical services. However, it may face several security threats and attacks over an insecure public network. Therefore, to protect sensitive medical data in IoMT, it is necessary to design a secure and efficient authentication protocol. In this study, we propose a privacy-preserving authentication management protocol based on blockchain. The protocol uses a blockchain to store identities and related parameters to assist communication entities in authentication. In addition, the protocol adopts a three-factor authentication method and introduces Chebyshev chaotic map to ensure the security of user login and authentication. Formal security proof and analysis using the random oracle model and Burrows-Abadi-Needham logic show that the proposed protocol is secure. Moreover, we use informal security analysis to demonstrate that the protocol can resist various security attacks. The functional comparison shows that the protocol has high security. Through performance analysis and comparison with other protocols, the proposed protocol can increase computation overhead, communication overhead, and storage overhead by up to 39.8%, 93.6%, and 86.7%,respectively.

H2O2-Mediated Synthesis of a Quinazolin-4(3H)-one Scaffold: A Sustainable Approach.

A quinazolin-4(3H)-one ring system is a privileged heterocyclic moiety with distinctive biological properties. From this perspective, the development of an efficient strategy for the synthesis of quinazolin-4(3H)-one has always been in demand for the synthetic chemistry community. In this report, we envisaged an efficient protocol for the synthesis of quinazolin-4(3H)-one using substituted 2-amino benzamide with dimethyl sulfoxide (DMSO) as a carbon source and H2O2 as an effective oxidant. Mechanistically, the reaction proceeds through the radical approach with DMSO as one carbon source. To further substantiate the synthetic claim, the synthetic protocol has been extended to the synthesis of the anti-endotoxic active compound 3-(2-carboxyphenyl)-4-(3H)-quinazolinone.

A facile catalyst-free one-pot three component synthesis of pharmacologically important indole-centered 4H-chromenes in a deep eutectic solvent (DES)

Popularity of the multicomponent reactions (MCRs) lies in the versatility as well as simplicity of their synthetic procedures which unlock the access to a variety of valuable bioactive products as well as fine chemicals by means the manifold prospects of the reagent combinations. In this paper, we report a simple, green, and highly efficient protocol to assemble diverse indole-based 4H-chromenes in good yields using a low melting mixture of choline chloride and urea. Herewith, we have employed inexpensive and easily accessible indole derivatives, salicylaldehyde, 2-hydroxy-1-naphthaldehyde, malononitrile, and ethyl cyanoacetate to prepare a variety of indole-centered 4H-chromenes. Importantly, the reaction medium was recycled/reused without any noticeable decrease in both yield as well as the catalytic activity. More importantly, the deep eutectic medium play a dual role, solvent as well as catalyst, and hence avoid the usage of volatile organic solvent and corrosive catalyst, as well. Moreover, we have also revealed the preliminary photophysical (absorption and emission) properties of thus prepared molecules. Since this particular method involve both green reaction medium as well as straightforward catalyst-free multicomponent procedure, it may open up new possibilities to synthesize a wide variety of hybrid indole and chromene based heterocycles of therapeutic uses under green and mild conditions in many years to come.

Sustainable energy and waste management: How to transform plastic waste into carbon nanostructures for electrochemical supercapacitors

Plastic waste consumption increases exponentially every year, mainly in the last three years due to the COVID-19 pandemic. The rapid growth of plastic products has exceeded the world’s capacity to deal with this type of trash. Thus, it has become a substantial environmental concern in modern society. Another dire concern is the improper disposal of used supercapacitors, leading to serious environmental impacts. Consequently, critical action to tackle this issue is to transform trash into high-valued materials, such as carbon nanomaterial supercapacitors. Considering several methodologies of recycling, pyrolysis stands out due to its simplicity and easy handling of mixed plastic waste to produce carbonaceous materials with different dimensions (0, 1, 2, and 3D). Thus, from this technology, it is possible to create new opportunities for using plastic waste and other types of waste to produce cheaper carbon-based materials for supercapacitors. This review aims to provide readers with a sustainability-driven view regarding the reutilization of plastic trash, discusses the environmental consequences of not doing so, and shows plastic waste solutions. Despite the broad scope of the topic, this review focuses on identifying the currently studied strategies to convert plastic waste into carbon-based electrodes, using less expensive and more efficient competitive protocols, besides emphasizing the diverse types (0, 1, 2, and 3D) of nanostructures. This review also proposes promising options for a sustainable cycle of plastic waste and supercapacitor.

Kinetic resolution of substituted amido[2.2]paracyclophanes via asymmetric electrophilic amination

Planar chiral [2.2]paracyclophane derivatives are a type of structurally intriguing and practically useful chiral molecules, which have found a range of important applications in the field of asymmetric catalysis and material science. However, access to enantioenriched [2.2]paracyclophanes represents a longstanding challenge in organic synthesis due to their unique structures, which are still highly dependent on the chiral chromatography separation technique and classical chemical resolution strategy to date. In this work, we report on an efficient and versatile kinetic resolution protocol for various substituted amido[2.2]paracyclophanes, including those with pseudo-geminal, pseudo-ortho, pseudo-meta and pseudo-para disubstitutions, using chiral phosphoric acid (CPA)-catalyzed asymmetric amination reaction, which was also applicable to the enantioselective desymmetrization of an achiral diamido[2.2]paracyclophane. Detailed experimental studies shed light on a new reaction mechanism for the electrophilic aromatic C-H amination, which proceeded through sequential triazane formation and N[1,5]-rearrangement. The facile large-scale kinetic resolution reaction and diverse derivatizations of both the recovered chiral starting materials and the C-H amination products showcased the potential of this method.

Easy cleaning plus stable activation of glassy carbon electrode surface by oxygen plasma

Glassy carbon (GC) electrodes are widely used in electroanalytical applications especially in bioelectrochemistry. Their use starts with an efficient surface cleaning and activation protocol, mostly based on surface polishing steps. We studied the use of an oxygen plasma exposure of GC electrodes to replace common polishing procedures. The cyclic voltammetry (CV) responses of ferrocyanide and ferrocene-dimethanol were used to compare brand new, surface-polished and plasma-treated GC electrodes. Plasma treatment induces CV responses with improved features, close to theoretical values, as compared to other methods. The plasma effects were quasi-stable over a week when electrodes were stored in water, this being explained by increased surface energy and hydrophilicity. Furthermore, when electroreduction of diazonium was performed on GC electrodes, the surface blockade could be removed by the plasma. Thus, a short oxygen plasma treatment is prone to replace polishing protocols, that display person-dependent efficiency, in most of the experiments with GC electrodes.

True-to-typeness and phytomedicinal potential in somatic embryo-derived plants of Crinum malabaricum (Amaryllidaceae): A medicinally important source of pharmaceutical biomolecules

Crinum malabaricum, Lekhak & S.R. Yadav (Amaryllidaceae) is a critically endangered bulbous plant endemic to India. Establishing an efficient plant regeneration protocol is necessary for its conservation and large-scale propagation. In the present study, regeneration was achieved through somatic embryogenesis from callus produced on MS media supplemented with various different concentrations of 2,4-D alone and in combination with N6-benzyl-adenine (BA). Different advanced stages of embryo development (globular, torpedo and cotyledonary) and maturation were obtained on MS medium, with combinations of picloram and thidiazuron (TDZ). A high rate of somatic embryos (55.89 ± 0.60) was obtained after eight weeks. SEM examinations showed the occurrence of cell clusters (embryogenic) which converted to somatic embryos. Well-developed cotyledon embryos were successfully germinated on PGRs free full and half strength MS medium, however, 94.03% of somatic embryos germinated on half-strength MS medium supplemented with 1.0 mg L−1 gibberellic acid. The true-to-type genetic conformity of regenerated plants was examined by SCoT, ISSR and RAPD primers based molecular analysis. The amplification bands produced by SCoT, ISSR and RAPD primers applied were monomorphic across all the regenerated plants. This confirmed their genetic homogeneity compared to the mother plant and also demonstrated the reliability of our somatic embryogenesis system for C. malabaricum. A rapid method for phytochemical analysis based on LC-ESI/MS exhibited better separation and analysis of galanthamine and lycorine from methanolic extracts of in vitro raised plants derived from somatic embryogenesis. The protocol developed should be helpful in reintroduction, restoration and ex situ conservation of this valuable plant in the natural condition as well as in the pharmaceutical sectors.