Smart reprograming of plants against salinity stress using modern biotechnological tools.

Climate change gives rise to numerous environmental stresses, including soil salinity. Salinity/salt stress is the second biggest abiotic factor affecting agricultural productivity worldwide by damaging numerous physiological, biochemical, and molecular processes. In particular, salinity affects plant growth, development, and productivity. Salinity responses include modulation of ion homeostasis, antioxidant defense system induction, and biosynthesis of numerous phytohormones and osmoprotectants to protect plants from osmotic stress by decreasing ion toxicity and augmented reactive oxygen species scavenging. As most crop plants are sensitive to salinity, improving salt tolerance is crucial in sustaining global agricultural productivity. In response to salinity, plants trigger stress-related genes, proteins, and the accumulation of metabolites to cope with the adverse consequence of salinity. Therefore, this review presents an overview of salinity stress in crop plants. We highlight advances in modern biotechnological tools, such as omics (genomics, transcriptomics, proteomics, and metabolomics) approaches and different genome editing tools (ZFN, TALEN, and CRISPR/Cas system) for improving salinity tolerance in plants and accomplish the goal of "zero hunger," a worldwide sustainable development goal proposed by the FAO.

An IoMT-Based Melanoma Lesion Segmentation Using Conditional Generative Adversarial Networks.

Currently, Internet of medical things-based technologies provide a foundation for remote data collection and medical assistance for various diseases. Along with developments in computer vision, the application of Artificial Intelligence and Deep Learning in IOMT devices aids in the design of effective CAD systems for various diseases such as melanoma cancer even in the absence of experts. However, accurate segmentation of melanoma skin lesions from images by CAD systems is necessary to carry out an effective diagnosis. Nevertheless, the visual similarity between normal and melanoma lesions is very high, which leads to less accuracy of various traditional, parametric, and deep learning-based methods. Hence, as a solution to the challenge of accurate segmentation, we propose an advanced generative deep learning model called the Conditional Generative Adversarial Network (cGAN) for lesion segmentation. In the suggested technique, the generation of segmented images is conditional on dermoscopic images of skin lesions to generate accurate segmentation. We assessed the proposed model using three distinct datasets including DermQuest, DermIS, and ISCI2016, and attained optimal segmentation results of 99%, 97%, and 95% performance accuracy, respectively.

Programmable Biosensors Based on RNA-Guided CRISPR/Cas Endonuclease.

Highly infectious illnesses caused by pathogens constitute severe threats to public health and lead to global economic loss. The use of robust and programmable clustered regularly interspaced short palindromic repeat and CRISPR-associated protein (CRISPR-Cas) systems, repurposed from genome-engineering applications has markedly improved traditional nucleic acid detection for precise identification, independently enabling rapid diagnostics of multiplex biomarker with genetic and mutation related to tumors, and microbial pathogens. In this review, we delineate the utility of the current CRISPR-Cas enzyme as biosensors by which these effector toolkits achieve recognition, signaling amplification, and finally, accurate detection. Additionally, we discuss the details of the dominance and hurdles related to expanding this revolutionary technology into an effective and convenient contraption crucial for improving the rational redesign to CRISPR/Cas biosensing. Overall, this review provides an insight into the current status of rapid and POC diagnostic systems by CRISPR/Cas tools.

Omics: a tool for resilient rice genetic improvement strategies.

Rice is pivotal pyramid of about half of the world population. Bearing small genome size and worldwide utmost food crop rice has been known as ideal cereal crop for genome research. Currently, decreasing water table and soil fatigue are big challenges and intense consequences in changing climate. Whole sequenced genome of rice sized 389 Mb of which 95% is covered with excellent mapping order. Sequenced rice genome helps in molecular biology and transcriptomics of cereals as it provides whole genome sequence of indica and japonica sub species. Through rice genome sequencing and functional genomics, QTLs or genes, genetic variability and halophyte blocks for agronomic characters were identified which have proved much more useful in molecular breeding and direct selection. There are different numbers of genes or QTLs identified for yield related traits i.e., 6 QTLs/genes for plant architecture, 6 for panicle characteristics, 4 for grain number, 1 gene/QTL for tiller, HGW, grain filling and shattering. QTLS/genes for grain quality, biotic stresses and for abiotic stresses are 7, 23 and 13 respectively. Low yield, inferior quality and susceptibility to biotic and abiotic stresses of a crop is due to narrow genetic background of new evolving rice verities. Wild rice provides genetic resources for improvement of these characters, molecular and genomics tool at different stages can overcome these stresses and improve yield and quality of rice crop.

LR-AKAP: A Lightweight and Robust Security Protocol for Smart Home Environments.

For the betterment of human life, smart Internet of Things (IoT)-based systems are needed for the new era. IoT is evolving swiftly for its applications in the smart environment, including smart airports, smart buildings, smart manufacturing, smart homes, etc. A smart home environment includes resource-constrained devices that are interlinked, monitored, controlled, and analyzed with the help of the Internet. In a distributed smart environment, devices with low and high computational power work together and require authenticity. Therefore, a computationally efficient and secure protocol is needed. The authentication protocol is employed to ensure that authorized smart devices communicate with the smart environment and are accessible by authorized personnel only. We have designed a novel, lightweight secure protocol for a smart home environment. The introduced novel protocol can withstand well-known attacks and is effective with respect to computation and communication complexities. Comparative, formal, and informal analyses were conducted to draw the comparison between the introduced protocol and previous state-of-the-art protocols.

Ammonia stress influences intestinal histomorphology, immune status and microbiota of Chinese striped-neck turtle (Mauremys sinensis).

Ammonia is one of major pollutants in aquatic environment that induces severe stress and toxicity to organisms in aquatic system. The intestine acts a major defense line that protects living organisms from biotic and abiotic stresses. In the current study, we examined the effects of ammonia on intestinal histomorphology, transcriptional levels of intestinal barrier functioning genes and intestinal microbiota of Chinese striped-neck turtle (Mauremys sinensis). Thus, the turtles were placed in water with addition of ammonia at 0 (control), 100, 200 mg L-1 for 30 days. Our findings showed that ammonia reduced the villus length and induced the inflammatory cells appearance. In addition, the epithelial tight junction genes, claudin and zonola occludin significantly downregulated in ammonia exposed groups as compared to control group (P < 0.05). Similarly, the mRNA expression levels of MUC-2 gene also significantly decreased in ammonia treated groups (P < 0.05). However, the expression levels of intestinal immune related genes such as IL-10, IL-12, TGF-ß1, TNF-α and IFN-γ significantly increased (P < 0.05). Furthermore, ammonia changed gut microbial diversity variedly. At the phylum levels, Firmicutes increased, whereas Bacteroidota, Desulfobacterota and Synergistota decreased significantly. Likewise, Lachnospiraceae, Bacteroides, Eubacteriaceae, Desulfovibrio, Muribaculaceae, Bilophila, Cloacibacillus, Christensenellaceae, Ruminococcus and Parabacteroides decreased while, Romboutsia and Turicibacter increased in ammonia exposed groups. In conclusion, ammonia at 100 and 200 mg L-1 could alter the intestinal barrier function and change the composition of intestinal microbiota, leading to bad health status in M. sinensis.

HEP-Net opinion on the management of ascites and its complications in the setting of decompensated cirrhosis in the resource constrained environment of Pakistan.

Approximately one half of patients develop ascites within 10 years of diagnosis of compensated cirrhosis. It is a poor prognostic indicator, with only 50% surviving beyond two years. Mortality worsens significantly to 20% to 50% at one year if the ascites becomes refractory to medical therapy. Pakistan has one of the highest prevalence of viral hepatitis in the world and patients with ascites secondary to liver cirrhosis make a major percentage of both inpatient and outpatient burden. Studies indicate that over 80% of patients admitted with ascites have liver cirrhosis as the cause. This expert opinion suggests proper assessment of patients with ascites in the presence of underlying cirrhosis. This expert opinion includes appropriate diagnosis and management of uncomplicated ascites, refractory ascites and complicated ascites (including spontaneous bacterial peritonitis (SBP) ascites, hepatorenal syndrome (HRS) and hyponatremia. The purpose behind this expert opinion is to help consultants, postgraduate trainees, medical officers and primary care physicians optimally manage their patients with cirrhosis and ascites in a resource constrained setting as is often encountered in a developing country like Pakistan.

Significant Risk of Graft-versus-Host Disease with Exposure to Checkpoint Inhibitors before and after Allogeneic Transplantation.

Investigators are using checkpoint inhibitors (CPIs) to treat aggressive hematologic malignancies in patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT) and in some patients with relapsed disease after allo-HSCT. CTLA-4 inhibitors and PD-1 inhibitors are 2 main types of CPIs, which work through activation of the immune system. On one hand, CPIs can achieve graft-versus-tumor effect, and on the other hand, there is a risk of graft-versus-host disease (GVHD). After a comprehensive literature review, we included data (n = 283) from 24 studies (11 original manuscripts and 13 case reports or case series) and evaluated the results to assess the safety and efficacy of CPI use in conjunction with allo-HSCT. Among the 283 patients, 107 received CPI before allo-HSCT, and 176 received CPI after allo-HSCT. The most common indication for CPI use was for Hodgkin lymphoma. The CPIs used in various studies included ipilimumab, nivolumab, and pembrolizumab. Among the patients exposed to CPI before allo-HSCT, 56% developed acute GVHD and 29% developed chronic GVHD. Investigators reported 20 deaths, 60% of which were GVHD-related. The overall mortality risk with GVHD is 11%. In this group, investigators noted an objective response rate (ORR) in 68% of patients, with complete remission (CR) in 47%, partial remission (PR) in 21%, and stable disease in 11%. Among the patients who received a CPI after allo-HSCT for disease relapse, 14% developed acute GVHD and 9% developed chronic GVHD. Investigators reported 40 deaths, 28% of which were GVHD-related. The mortality risk with GVHD is approximately 7%. Investigators reported ORR in 54% of patients, with CR in 33%, PR in 21%, and disease stabilization in 5%. After careful evaluation of collective data, we found that CPI use both before and after allo-HSCT can be highly effective, but exposure can lead to a significantly increased risk of GVHD-related morbidity and mortality in this patient population. Despite limited availability of data, there is need for extreme caution while making decisions regarding the use of CPIs. Detailed discussions and prospective well-designed clinical trials are needed to explore this issue further.

General Approach to Produce Nanostructured Binary Transition Metal Selenides as High-Performance Sodium Ion Battery Anodes.

Multiple transition metals containing chalcogenides have recently drawn boosted attraction as anodes for sodium ion batteries (SIBs). Their greatly enhanced electrochemical performances can be attributed to the superior intrinsic conductivities and richer redox reactions, comparative to mono metal chalcogenides. To employ various binary metals comprising selenides (B-TMSs) for SIBs, discovery of a simplistic, scalable and universal synthesis approach is highly desirable. Herein, a simple, facile, and comprehensive strategy to produce various combinations of nanostructured B-TMSs is presented. As a proof of concept, optimized, high surface area bearing, and hierarchical nanosheets of iron-nickel selenide (FNSe), iron-cobalt selenide, and nickel-cobalt selenide are produced and employed in SIBs. These B-TMSs exhibit adequately high energy capacities, excellent rate capabilities, and an extraordinarily stable life of 2600 cycles. As far as it is known, it is the first work to discuss sodium storage of FNSe, so various in situ and ex situ battery analyses are carried out to probe the sodium storage mechanism. When employed in sodium full batteries, these B-TMSs present reasonably high reversible specific capacities even after 100 cycles. Overall, the presented strategy will pave the way for facile synthesis of numerous binary transition metal chalcogenides that are the potential materials for energy storage and conversion systems.

Expression of recombinant tachyplesin I in Pichia pastoris.

The development of antibiotic-resistant bacteria has become a major public health problem, prompting the search for alternative solutions. Tachyplesin I (TP-I) is an antimicrobial peptide, which exhibits potent and broad-spectrum activities against bacteria, fungi, viruses, and tumor cells. However, limited amounts of TP-I produced in horseshoe crab restrict its large-scale use. In order to solve this problem, a eukaryotic expression system of Pichia pastoris with high TP-I expression was constructed by gene engineering. To achieve high expression of TP-I, 74 amino acid-long peptide (4TP-1) was designed containing 4 copies of TP-I, and specific cleavage sites for pancreatic elastase (-Ala↓ or -Gly↓) and carboxypeptidase A (cleaves C terminal amino acid); these cleavage sites for enzymes were located between the four copies of TP-I. The gene sequence for the designed peptide was synthesized taking into consideration codon preferences for P. pastoris, and cloned into the highly efficient expression vector pGAPZα B. Host Pichia pastoris strain GS115 cells were transfected by the constructed expression vector pGAPZα B-4tp-I by electroporation. Tricine-SDS-PAGE electrophoresis was carried out to detect the expression of target peptides in the fermentation medium. This analysis showed a protein band of 3.3 kDa, identical to that of chemically synthesized TP-I, verifying that successful synthesis and secretion of TP-I by genetically engineered P. pastoris. The concentration of TP-I in the fermentation broth was 27.24-29.53 mg/L. High-resolution mass spectrometry analysis documented that the TP-I monomer had the same molecular weight, 2262.85, as the designed 17-amino acid sequence. The recombinant TP-I peptide displayed different levels of bactericidal activity against Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa, and Staphylococcus aureus. Thus, the present study demonstrated the feasibility of achieving high levels of expression of TP-I in P. pastoris.

The Role of Nanotechnology in Food Safety: Current Status and Future Perspective.

Nanotechnology is an emerging science in food production and processing sector, yet the role of nanotechnology in food safety has not been comprehensively reviewed. This study reviewed the types, sources and mode of actions of the nanoparticles used in the food systems. Additionally, the effect of nanoparticles on animal health and safety of the products of animal origin was evaluated. Moreover, retention of nutritionally important nanoparticle minerals in the animal systems and foods of animal origins was analyzed. Furthermore, food safety was critically evaluated in terms of antioxidative ability, antibacterial properties, and toxicological studies. Finally, the scope of nanoparticle-based functional foods and shelf-life enhancement using active packaging was discussed. The article concluded that although significant research has been done on the use of nanoparticles in food systems, yet commercialization of nanoparticle-based foods needs further investigation.

A Review on NanoPCR: History, Mechanism and Applications.

Polymerase Chain Reaction (PCR) is one of the most common technologies used in many laboratories to produce millions of copies of targeted nucleic acid under in vitro conditions. However, PCR faces multiple challenges including limited availability of DNA in the sample, high GC contents of the template, low efficiency, and specificity in amplification. Moreover, some DNA fragments are very difficult to amplify due to their secondary structure and high melting temperature requirement. To overcome these challenges, many approaches including the application of PCR additives in PCR mixture; change in instrument design; optimization of PCR system by using the accurate concentration of magnesium ions, primers, and cycle number; enzyme modification; and setting up the new touchdown and nested PCR strategies have been adopted. Although these approaches have enriched the output of PCR, they are not all-purpose and optimization can be case dependent. Nanometer-sized materials (nanomaterials) have offered a possible solution to these problems as these materials have exceptional physio-chemical properties as compared to macroscopic materials. Among these nanomaterials, silicon-based materials, carbon-based materials, semiconductor quantum dots (QDs), and some metals are well-known PCR enhancer. Hence, new PCR has been designed to utilize the unique properties of nanomaterial and is known as nanomaterial-assisted PCR or simply nanoPCR. Results of many studies have shown that the combination of these nanomaterials and biomolecules can mimic the DNA replication process successfully as present in the living organism. In this review, we have discussed the role of these different nanomaterials one by one and also discussed the mechanisms through which these nanomaterials enhance the efficiency of PCR.

Animal research for type 2 diabetes mellitus, its limited translation for clinical benefit, and the way forward.

Obesity and type 2 diabetes mellitus (T2DM) have reached pandemic proportions worldwide, and considerable research efforts have been dedicated to investigating disease pathology and therapeutic options. The two hallmark features of T2DM, insulin resistance and pancreatic dysfunction, have been studied extensively by using various animal models. Despite the knowledge acquired from such models, particularly mechanistic discoveries that sometimes mimic human T2DM mechanisms or pathways, many details of human T2DM pathogenesis remain unknown, therapeutic options remain limited, and a cure has eluded research. Emerging human data have raised concern regarding inter-species differences at many levels (e.g. in gene regulation, pancreatic cytoarchitecture, glucose transport, and insulin secretion regulation), and the subsequent impact of these differences on the clinical translation of animal research findings. Therefore, it is important to recognise and address the translational gap between basic animal-based research and the clinical advances needed to prevent and treat T2DM. The purpose of this report is to identify some limitations of T2DM animal research, and to propose how greater human relevance and applicability of hypothesis-driven basic T2DM research could be achieved through the use of human-based data acquisition at various biological levels. This report addresses how in vitro, in vivo and in silico technologies could be used to investigate particular aspects of human glucose regulation. We do not propose that T2DM animal research has been without value in the identification of mechanisms, pathways, or potential targets for therapies, nor do we claim that human-based methods can provide all the answers. We recognise that the ultimate goal of T2DM animal research is to identify ways to advance the prevention, recognition and treatment of T2DM in humans, but postulate that this is where the use of animal models falls short, despite decades of effort. The best way to achieve this goal is by prioritising human-centred research.

Integrated Design of MnO2 @Carbon Hollow Nanoboxes to Synergistically Encapsulate Polysulfides for Empowering Lithium Sulfur Batteries.

Lithium sulfur batteries (LSBs) with high theoretical energy density are being pursued as highly promising next-generation large-scale energy storage devices. However, its launch into practical application is still shackled by various challenges. A rational nanostructure of hollow carbon nanoboxes filled with birnessite-type manganese oxide nanosheets (MnO2 @HCB) as a new class of molecularly-designed physical and chemical trap for lithium polysulfides (Li2 Sx (x = 4-8)) is reported. The bifunctional, integrated, hybrid nanoboxes overcome the obstacles of low sulfur loading, poor conductivity, and redox shuttle of LSBs via effective physical confinement and chemical interaction. Benefiting from the synergistic encapsulation, the developed MnO2 @HCB/S hybrid nanoboxes with 67.9 wt% sulfur content deliver high specific capacity of 1042 mAh g-1 at the current density of 1 A g-1 with excellent Coulombic efficiency ≈100%, and retain improved reversible capacity during long term cycling at higher current densities. The developed strategy paves a new path for employing other metal oxides with unique architectures to boost the performance of LSBs.

Simultaneous Extraction of DNA and RNA from Hepatocellular Carcinoma (Hep G2) Based on Silica-Coated Magnetic Nanoparticles.

Nucleic acid (NA) extraction from cancer cells is an essential step in molecular oncologic testing. The conventional NA extraction protocols, based on several ultracentrifugation steps, suffer from time-consuming and complex manipulation. Here, a magnetic nanoparticle (MNP) based method for simultaneous extraction of DNA and RNA from cancer cells is described. This MNP based technique has received great attention and significant interest due to its convenient manipulation, low cost and ease for automation. Different factors including lysis buffer, ethanol, MNPs and washing buffers which may affect the yield of nucleic acid were optimized. The average yield of DNA and RNA obtained from 1 mL Hep G2 (˜106 cells) ranged from 9.7 to 14.7 µg with A260/A280 values between 1.68 and 2.01. The isolated DNA and RNA, using this method, were suitable for downstream activities such as PCR and RT-PCR.

Integration of Nucleic Acid Extraction Protocol with Automated Extractor for Multiplex Viral Detection.

The isolation of nucleic acids (NA) is the preliminary step to carry out genetic studies and DNA biosensor development. The presence of inhibitors in the purified NA interferes with the downstream application. These salts and other organic contaminations particularly challenge the analytical sensitivity of DNA biosensors. The detailed study was carried out to optimize the factors which might affect viral nucleic acid purification. The results suggested that 6 M guanidinium hydrochloride salt concentration was critical for NA isolation. The inverse relation has been found in the pH of the lysis buffer and quality and quantity of NA. The NA yield was relatively stable at pH 4­5. It has been observed that the use of carrier RNA was indispensable for viral genome isolation. The addition of ethanol to lysate in 1:1 ratio greatly improved NA recovery. The elution efficiency of DNase and RNase free water, 1× TE buffer and 1× PCR buffer was compared. The carrier RNA was best eluted in DNase and RNase free water and 1× TE buffer. It was further demonstrated that this method can be automatized for high throughput detection. A simple experiment was conducted to optimize the different parameters of an automated NA extractor to simultaneously extract HBV DNA and HCV RNA. The purified NA was successfully amplified in PCR and RT-PCR to verify the reliability of the established protocol. Thus a semi-automated system for the simultaneous detection of multiple viruses has been demonstrated.

Applications of Magnetic Nanoparticles in Targeted Drug Delivery System.

Magnetic nanoparticles (MNPs) are a special kind of nanomaterials and widely used in biomedical technology applications. Currently they are popularly customized for disease detection and treatment, particularly as drug carriers in drug targeted delivery systems, as a therapeutic in hyperthermia (treating tumors with heat), and as contrast agents in magnetic resonance imaging (MRI). Due to their biocompatibility and superparamagnetic properties, MNPs as next generation drug carriers have great attraction. Although the potential benefits of MNPs are considerable, any potential toxicity associated with these MNPs should be identified distinctly. The drug loading capability and the biomedical properties of MNPs generated by different surface coatings are the most sensitive parameters in toxicity. A lot of organic and inorganic materials are utilized as coating materials for surface functionalization and reducing toxicity of MNPs. pH or temperature sensitivity materials are widely used to manage drug loading and targeted release. In addition, MNPs can be controlled and directed to the desired pathological region by using external magnetic files (EMF). The realization of targeted drug delivery has decreased the dosage and improved the efficiency of drugs, which results in reduced side effects to normal tissues. This review discussed the possible organ toxicities of MNPs and their current advances as a drug delivery vehicle.

DNA Mediated Vaccines Delivery Through Nanoparticles.

Vaccination has led to the eradication of those diseases which had once claimed millions of lives worldwide; however, it is accompanied with a number of dis-advantages especially safety issues until the entry of DNA vaccines. The DNA vaccines have been emerged as the best remedy for problematic diseases being capable of producing humoral and cellular immune responses as well as the safest vaccines so far. However, the magnitude of immune responses produced in primates is lower than that in experimental animals. There are several reasons described theoretically for this limited efficacy and a number of novel approaches have been applied to boost their immune responses, e.g., use of more efficient promoters and coding optimization, addition of traditional or genetic adjuvants, electroporation, intradermal delivery and various prime-boost strategies. One of these strategies is controlled antigen administration of plasmid DNA through microspheres and nanoparticles. This approach is accompanied with a number of advantages to overcome the limitations of traditional delivery systems in terms of stability, solubility and pharmacology. Furthermore, the surface structure of a virus highly resembles with a nanoparticle because of their geometrical regularities and nanoscale dimensions; therefore, the engineering of nanoparticles is based upon principles of natural virus attack which will be the best tool for vaccination. There is evidence that these immune responses can be augmented by properly structured nanosized particles (nanoparticles) that may avoid DNA degradation and facilitate targeted delivery to antigen presenting cells. Adsorption, formulation or encapsulation with particles has been found to stabilize DNA formulations. The use of nanoparticles for DNA vaccine delivery is a platform technology and has been applied for delivery of a variety of existing and potential vaccines successfully.

ALT CHANGES AND ADVERSE EVENTS OF TELBIVUDINE IN HEPATITIS-B PATIENTS-AN EXPERIENCE OF 11 PATIENTS.

Hepatitis-B virus (HBV) infection is a major global health problem. Of the two billion people who have been infected, more than 350 million have chronic hepatitis. It is estimated that 235,000-328,000 people die annually due to liver cirrhosis and hepatocellular carcinoma, we assessed the short term outcomes of treatment with telbivudine in 11 adults aged 14-41 years with HBeAg-positive or HBeAg-negative chronic hepatitis-B (CHB). Treatment of chronic hepatitis-B patients with telbivudine shows 43.1% reduction in serum ALT with no significant adverse effects.