Bacterial tyrosinases and their inhibitors.

Bacterial tyrosinase is a copper-containing metalloenzyme with diverse physio-chemical properties, that have been identified in various bacterial strains, including actinobacteria and proteobacteria. Tyrosinases are responsible for the rate-limiting catalytic steps in melanin biosynthesis and enzymatic browning. The physiological role of bacterial tyrosinases in melanin biosynthesis has been harnessed for the production of coloring and dyeing agents. Additionally, bacterial tyrosinases have the capability of cross-linking activity, demonstrated material functionalization applications, and applications in food processing with varying substrate specificities and stability features. These characteristics make bacterial tyrosinases a valuable alternative to well-studied mushroom tyrosinases. The key feature of substrate specificity of bacterial tyrosinase has been exploited to engineer biosensors that have the ability to detect the minimal amount of different phenolic compounds. Today, the world is facing the challenge of multi-drugs resistance in various diseases, especially antibiotic resistance, skin cancer, enzymatic browning of fruits and vegetables, and melanogenesis. To address these challenges, medicinal scientists are developing novel chemotherapeutic agents by inhibiting bacterial tyrosinases. To serve this purpose, heterocyclic compounds are of particular interest due to their vast spectrum of biological activities and their potential as effective tyrosinase inhibitors. In this chapter, a plethora of research explores applications of bacterial tyrosinases in different fields, such as the production of dyes and pigments, catalytic applications in organic synthesis, bioremediation, food and feed applications, biosensors, wool fiber coating and the rationalized synthesis, and structure-activity relationship of bacterial tyrosinase inhibitors.

Diagnostic Accuracy of Polymerase Chain Reaction Against Giemsa Staining on Tissue Biopsy for Cutaneous Leishmaniasis.

OBJECTIVE: To evaluate the diagnostic accuracy of a commercial real-time polymerase chain reaction (PCR) kit targeting 18S rRNA against Giemsa-stained tissue slides in patients clinically suspected of cutaneous leishmaniasis (CL). STUDY DESIGN: Cross-sectional analytical study. Place and Duration of the Study: Department of Microbiology, Armed Forces Institute of Pathology / National University of Medical Sciences, Rawalpindi, Pakistan, from July to December 2022. METHODOLOGY: Samples of skin tissue in 98 patients suspected of CL were evaluated. These samples were subjected to Giemsa-staining for microscopy and real-time PCR. Sensitivity, specificity, and accuracy of the PCR were calculated keeping Giemsa-stained tissue slide microscopy as gold standard. RESULTS: Out of the 98 tissue samples, 37 were found positive for leishmaniasis on PCR while 13 were found Leishmania positive on microscopy of Giemsa-stained slides. The sensitivity, specificity, and accuracy of the PCR for the detection of Leishmania species were 100%, 71.8%, and 91.8%, respectively with 100% negative predictive value. CONCLUSION: This study demonstrates that the commercial PCR is a reliable diagnostic test for the diagnosis of CL. The ease, rapidity, and reliability of the PCR make it a dependable tool in diagnostic repertoire of CL. KEY WORDS: Giemsa stain, Leishmania spp., Polymerase chain reaction, Viasure.

Multitargeted molecular docking and dynamics simulation studies of 1,3,4-thiadiazoles synthesised from (R)-carvone against specific tumour protein markers: An In-silico study of two diastereoisomers.

In the present work, we describe the synthesis of new 1,3,4-thiadiazole derivatives from natural (R)-carvone in three steps including, dichloro-cyclopropanation, a condensation with thiosemicarbazide and then a 1,3-dipolar cycloaddition reaction with various nitrilimines. the targeted compounds were structurally identified by 1H & 13C NMR and HRMS analyses. The cytotoxic assay demonstrated that some synthesized novel compounds were potent on certain cancer cell lines. Molecular modeling studies were undertaken to rationalize the wet lab study results. Furthermore, molecular docking was performed to unveil the binding potential of the most active derivatives, 3a and 6c, to caspase-3 and COX-2. The stabilities of the protein-compound complexes obtained from the docking were evaluated using MD simulation. Furthermore, FMO and related parameters of the active compounds and their stereoisomers were examined through DFT studies. The docking study showed compound 6c had a higher binding potential than caspase-3. However, the binding strength of 6c was found to be less than that of the standard drug, doxorubicin, as it formed lower conventional hydrogen bonds. On the other hand, compound 3a had a higher binding potential to COX-2. However, the binding potential 3a was much lower than that of the standard COX-2 inhibitor, celecoxib. The MD simulation demonstrated that the caspase-3-6c complex was less stable than the caspase-3-doxorubicin complex. In contrast, the COX-2-3a complex was stable, and 3a was anticipated to remain inside the protein's binding pocket. The DFT study showed that 3a had higher chemical stability than 6c. The electron exchange capacity, chemical stability, and molecular orbital distributions of the stereoisomers of the active compounds were also found to be alike.

Removal of oxytetracycline from pharmaceutical wastewater using kappa carrageenan hydrogel.

This study investigated the adsorption of Oxytetracycline (OTC) from pharmaceutical wastewater using a kappa carrageenan based hydrogel (KPB). The aim of the present study was to explore the potential of KPB for long-term pharmaceutical wastewater treatment. A sustainable adsorbent was developed to address oxytetracycline (OTC) contamination. The hydrogel's structural and adsorption characteristics were examined using various techniques like Scanning Electron Microscope (SEM), Fourier Transform Infrared (FTIR), X-ray powder diffraction (XRD), and kinetic models. The results revealed considerable changes in the vibrational modes and adsorption bands of the hydrogel, suggesting the effective functionalization of Bentonite nano-clay. Kappa carrageenan based hydrogel achieved the maximum removal (98.5%) of OTC at concerntration of 40 mg/L, pH 8, cotact time of 140 min and adsorbent dose of 0.1 g (KPB-3). Adsorption of OTC increased up to 99% with increasing initial concentrations. The study achieved 95% adsorption capacity for OTC using a KPB film at a concentration of 20 mg/L and a 0.1 g adsorbent dose within 60 min. It also revealed that chemisorptions processes outperform physical adsorption. The Pseudo-Second-Order model, which emphasized the importance of chemical adsorption in the removal process, is better suited to represent the adsorption behavior. Excellent matches were found that R2 = 0.99 for KPB-3, R2 = 0.984 for KPB-2 and R2 = 0.989 for KPB-1 indicated strong chemical bonding interactions. Statisctical analysis (ANOVA) was performed using SPSS (version 25) and it was found that pH and concentration had significant influence on OTC adsorption by the hydrogel, with p-values less than 0.05. The study identified that a Kappa carrageenan-based hydrogel with bentonite nano-clay and polyvinyl alcohol (PVA) can efficiently remove OTC from pharmaceutical effluent, with a p-value of 0.054, but weak positive linear associations with pH, temperature, and contact time. This research contributed to sustainable wastewater treatment and environmental engineering.

Genome wide identification of phenylalanine ammonia-lyase (PAL) gene family in Cucumis sativus (cucumber) against abiotic stress.

Phenylalanine ammonia lyase (PAL) is a widely studied enzyme in plant biology due to its role in connecting primary metabolism to secondary phenylpropanoid metabolism, significantly influencing plant growth, development, and stress response. Although PAL genes have been extensively studied in various plant species but their exploration in cucumber has been limited. This study successfully identified 11 CsPAL genes in Cucumis sativus (cucumber). These CsPAL genes were categorized based on their conserved sequences revealing patterns through MEME analysis and multiple sequence alignment. Interestingly, cis-elements related to stress were found in the promoter regions of CsPAL genes, indicating their involvement in responding to abiotic stress. Furthermore, these gene's promoters contained components associated with light, development and hormone responsiveness. This suggests that they may have roles in hormone developmental processes. MicroRNAs were identified as a key regulators for the CsPAL genes, playing a crucial role in modulating their expression. This discovery underscores the complex regulatory network involved in the plant's response to various stress conditions. The influence of these microRNAs further highlights the complicated mechanisms that plants use to manage stress. Gene expression patterns were analyzed using RNA-seq data. The significant upregulation of CsPAL9 during HT3h (heat stress for 3 h) and the heightened upregulation of both CsPAL9 and CsPAL7 under HT6h (heat stress for 6 h) in the transcriptome study suggest a potential role for these genes in cucumber's tolerance to heat stress. This comprehensive investigation aims to enhance our understanding of the PAL gene family's versatility, offering valuable insights for advancements in cucumber genetics.

Assessment of growth, and ion uptake of plant species, Conocarpus erectus and Dodonaea viscosa, on industrial solid waste.

Present study assessed the growth of two plant species and ion uptake by them grown on different proportion of industrial solid waste and garden soil. The industrial waste having high concentration of chemicals were used with garden soil at different proportion i.e. 0% (T0), 5% (T1), 10% (T2), 15% (T3) and 20% (T4). Two species namely Conocarpus erectus (alien plant) and Dodonaea viscosa (indigenous) were used as test plants in pot study. Different parameters including growth, physiology, and anatomy of plants and concentration of cations (Na+, K+, Ca2+, and Mg2+) in the plant shoot and root were measured at different time duration (initial, 1st, 2nd, 3rd and 4th month). The key objective of the study was to use these plants to establish their plantations on the barren lands where industrial solid wastes were being disposed of. C. erectus showed better growth than D. viscosa, as well as more uptake of ions. A significant increase in plant growth was observed in fourth month in T1, where plant height reached 24.5% and 46% for C. erectus and D. viscosa, respectively. At harvest, in C. erectus, no significant difference in the fresh (65-78 g) and dry weight (24-30 g) of the shoot was observed across treatments compared to the control. In D. viscosa, at the time of harvest, the fresh and dry weights of the root and shoot showed a strong, significantly decreasing pattern across T1, T2, and T3, leading to the death of the plant at T3 and T4. Further, optimum ratio of waste soil to garden soil was found as 10:90 and 20:80 to establish the plantations of D. viscosa and C. erectus, respectively in areas where such solid waste from industries are disposed. Findings can be used for the restoration of such solid waste for the sustainable management of industrial areas and their associated ecosystems.

Genome-scale identification, expression and evolution analysis of B-box members in Dendrobium huoshanense.

B-box (BBX) proteins have been recognized as vital determinants in plant development, morphogenesis, and adaptive responses to a myriad of environmental stresses. These zinc-finger proteins play a pivotal role in various biological processes. Their influence spans photomorphogenesis, the regulation of flowering, and imparting resilience to a wide array of challenges, encompassing both biotic and abiotic factors. Chromosome localization, gene structure and conserved motifs, phylogenetic analysis, collinearity analysis, expression profiling, fluorescence quantitative analysis, and tobacco transient transformation methods were used for functional localization and expression pattern analysis of the DhBBX gene. A total of 23 DhBBX members were identified from Dendrobium huoshanense. Subsequent phylogenetic evaluations effectively segregated these genes into five discrete evolutionary subsets. The predictions of subcellular localizations revealed that all these proteins were localized in the nucleus. The genetic composition and patterns showed that the majority of these genes consisted of several exons, with a few variations that could be attributed to transposon insertion. A comprehensive analysis using qRT-PCR was conducted to unravel the expression patterns of these genes in D. huoshanense, with a specific concentration on their responses to various hormone treatments and cold stress. Subcellular localization reveals that DhBBX21 and DhBBX9 are located in the nucleus. Our results provide a deep comprehension of the complex regulatory mechanisms of BBXs in response to various environmental and hormonal stimuli. These discoveries encourage further detailed and focused investigations into the operational dynamics of the BBX gene family in a wider range of plant species.

Pediatric penetrating brain injuries with intact neurological function: Two case reports of a pencil and a toy wheel encounter.

Accidental injuries in the pediatric population are common. The response to injury also differs owing to anatomical and physiological differences in children. While such injuries carry a risk of lifelong morbidity, some cases may follow a benign course despite their distressing appearance. We report two cases of accidentally incurred penetrating trauma in the pediatric population with unusual objects, including a pencil and a toy wheel. Despite their intracranial extension, neither of the patients exhibited any discernible neurological deficits. Penetrating brain injuries require early removal and meticulous perioperative care to minimize the risk of long-term adverse neurological events in children.

Genome-wide analysis and identification of Carotenoid Cleavage Oxygenase (CCO) gene family in coffee (coffee arabica) under abiotic stress.

The coffee industry holds importance, providing livelihoods for millions of farmers globally and playing a vital role in the economies of coffee-producing countries. Environmental conditions such as drought and temperature fluctuations can adversely affect the quality and yield of coffee crops.Carotenoid cleavage oxygenases (CCO) enzymes are essential for coffee plants as they help break down carotenoids contributing to growth and stress resistance. However, knowledge about the CCO gene family in Coffee arabica was limited. In this study identified 21 CCO genes in Coffee arabica (C. arabica) revealing two subfamilies carotenoid cleavage dioxygenases (CCDs) and 9-cis-epoxy carotenoid dioxygenases (NCED) through phylogenic analysis. These subfamilies exhibited distribution patterns in terms of gene structure, domains, and motifs. The 21 CaCCO genes, comprising 5 NCED and 16 CCD genes were found across chromosomes. Promoter sequencing analysis revealed cis-elements that likely interact with plant stress-responsive, growth-related, and phytohormones, like auxin and abscisic acid. A comprehensive genome-wide comparison, between C. arabica and A. thaliana was conducted to understand the characteristics of CCO genes. RTqPCR data indicated that CaNCED5, CaNCED6, CaNCED12, and CaNCED20 are target genes involved in the growth of drought coffee plants leading to increased crop yield, in a conditions, with limited water availability. This reveals the role of coffee CCOs in responding to abiotic stress and identifies potential genes useful for breeding stress-resistant coffee varieties.

Efficient chromium removal from leather industrial wastewater in batch experimental study: Green synthesis and characterization of zinc oxide nanoparticles using Ficus benghalensis extracts.

The urgent need to address the severe environmental risk posed by chromium-contaminated industrial wastewater necessitates the development of eco-friendly cleanup methodologies. Utilizing the Ficus benghalensis plant extracts, the present study aims to develop green zinc oxide nanoparticles for the removal of Cr metal ions from wastewater. The leaves of Ficus benghalensis, often known as the banyan tree, were used to extract a solution for synthesizing ZnO NPs. These nanoparticles were developed with the goal of efficiently eliminating chromium (Cr) from industrial effluents. Batch studies were carried out to assess the efficiency of these synthesized ZnO NPs in treating leather industrial effluent, with aiming for optimal chromium removal. This involved measuring the nanoparticles' capacity to adsorb Cr ions from wastewater samples by comparing chromium levels before and after treatment. Removal efficiency for Cr was estimated through the batches such as optimization of pH, contact time, initial Cr concentration and sorbent dose of ZnO NPs were of the batches. These synthesized ZnO NPs were found to be successful in lowering chromium levels in wastewater to meet permissible limit. The nanoparticles exhibited their highest absorption capacity, reaching 94 % (46 mg/g) at pH 4, with a contact time of 7 hours with the optimum sorbent dose of 0.6 g/L. Hence, the excellent adsorption capabilities of these nanoparticles, together with their environmentally benign manufacturing technique, provide a long-term and efficient solution for chromium-contaminated wastewater treatment. Its novel nature has the potential to significantly improve the safety and cleanliness of water ecosystems, protecting the both i.e. human health and the environment.

Assessment of drinking water quality using Water Quality Index and synthetic pollution index in urban areas of mega city Lahore: a GIS-based approach.

The aim of the present study was to assess the drinking water quality in the selected urban areas of Lahore and to comprehend the public health status by addressing the basic drinking water quality parameters. Total 50 tap water samples were collected from groundwater in the two selected areas of district Lahore i.e., Gulshan-e-Ravi (site 1) and Samanabad (site 2). Water samples were analyzed in the laboratory to elucidate physico-chemical parameters including pH, turbidity, temperature, total dissolved solids (TDS), electrical conductivity (EC), dissolved oxygen (DO), total hardness, magnesium hardness, and calcium hardness. These physico-chemical parameters were used to examine the Water Quality Index (WQI) and Synthetic Pollution Index (SPI) in order to characterize the water quality. Results of th selected physico-chemical parameters were compared with World Health Organization (WHO) guidelines to determine the quality of drinking water. A GIS-based approach was used for mapping water quality, WQI, and SPI. Results of the present study revealed that the average value of temperature, pH, and DO of both study sites were within the WHO guidelines of 23.5 °C, 7.7, and 6.9 mg/L, respectively. The TDS level of site 1 was 192.56 mg/L (within WHO guidelines) and whereas, in site 2 it was found 612.84 mg/L (higher than WHO guidelines), respectively. Calcium hardness of site 1 and site 2 was observed within the range from 25.04 to 65.732 mg/L but, magnesium hardness values were higher than WHO guidelines. The major reason for poor water quality is old, worn-out water supply pipelines and improper waste disposal in the selected areas. The average WQI was found as 59.66 for site 1 and 77.30 for site 2. Results showed that the quality of the water was classified as "poor" for site 1 and "very poor " for site 2. There is a need to address the problem of poor water quality and also raise the public awareness about the quality of drinking water and its associated health impacts.

Performance evaluation of E-VGG19 model: Enhancing real-time skin cancer detection and classification.

Skin cancer is a pervasive and potentially life-threatening disease. Early detection plays a crucial role in improving patient outcomes. Machine learning (ML) techniques, particularly when combined with pre-trained deep learning models, have shown promise in enhancing the accuracy of skin cancer detection. In this paper, we enhanced the VGG19 pre-trained model with max pooling and dense layer for the prediction of skin cancer. Moreover, we also explored the pre-trained models such as Visual Geometry Group 19 (VGG19), Residual Network 152 version 2 (ResNet152v2), Inception-Residual Network version 2 (InceptionResNetV2), Dense Convolutional Network 201 (DenseNet201), Residual Network 50 (ResNet50), Inception version 3 (InceptionV3), For training, skin lesions dataset is used with malignant and benign cases. The models extract features and divide skin lesions into two categories: malignant and benign. The features are then fed into machine learning methods, including Linear Support Vector Machine (SVM), k-Nearest Neighbors (KNN), Decision Tree (DT), Logistic Regression (LR) and Support Vector Machine (SVM), our results demonstrate that combining E-VGG19 model with traditional classifiers significantly improves the overall classification accuracy for skin cancer detection and classification. Moreover, we have also compared the performance of baseline classifiers and pre-trained models with metrics (recall, F1 score, precision, sensitivity, and accuracy). The experiment results provide valuable insights into the effectiveness of various models and classifiers for accurate and efficient skin cancer detection. This research contributes to the ongoing efforts to create automated technologies for detecting skin cancer that can help healthcare professionals and individuals identify potential skin cancer cases at an early stage, ultimately leading to more timely and effective treatments.

Aloe-inspired eco-friendly synthesis of Ag/ZnO heterostructures: boosting photocatalytic potential.

The current research focuses on the development of Ag-ZnO heterostructures through a "bottom-up" approach involving the assembly and extraction of Aloe barbadensis Miller gel. These heterostructures composed of metals/semiconductor oxide display distinct and notable optical, electrical, magnetic, and chemical properties that are not found in single constituents and also exhibit photocatalytic applications. These synthesized heterostructures were characterized by XRD, FTIR, SEM, and UV-visible spectroscopy. The high peak intensity of the Ag/ZnO composite shows the high crystallinity. The presence of Ag-O, Zn-O, and O-H bonding is verified using FTIR analysis. SEM analysis indicated the formation of spherical shapes of Ag/ZnO heterostructures. The Zn, O, and Ag elements are further confirmed by EDX analysis. Ag-ZnO heterostructures exhibited excellent photocatalytic activity and stability against the degradation of tubantin red 8BL dye under visible light irradiation.

A novel approach to assessing the antioxidant and anti-diabetic potential of synthesized calcium carbonate nanoparticles using various extracts of Ailanthus altissima.

Calcium carbonate nanoparticles (CaCO3) have been found to exhibit unique properties that show their potential to be used in various therapies. Green synthesis of CaCO3 has been progressively gaining ac-ceptance due to its cost-effectiveness and energy-efficient nature. In the current study, different extracts of Ailanthus altissima were used to synthesize the calcium carbonate nanoparticles the synthesis and characterization of CCNPs were confirmed by using Fourier Transform Infra-Red spectroscopy, UV-Vis spectroscopy, and Scanning Electron Microscopy (SEM). The antioxidant activities (hydrogen peroxide, phosphomolydbenum, and ferric reducing) of calcium carbonate nanoparticles were affirmed by a good range of percentages of inhibition against free radical scavenging. The antidebate assays of CCNPs were observed by in-vitro and in silico approaches in a range at various concentrations while maximum inhibition occurred. In conclusion, the current study depicted that conjugated CaCO3 with A. altissima has a good potential to cure oxidative stress and Type II diabetes and could be used in the future as biogenic nanomedicine for the treatment of other metabolic diseases.

Discovery of novel 1,2,4-triazole tethered ß-hydroxy sulfides as bacterial tyrosinase inhibitors: synthesis and biophysical evaluation through in vitro and in silico approaches.

In this study, a series of 1,2,4-triazole-tethered ß-hydroxy sulfide scaffolds 11a-h was synthesized in good to remarkable yields (69-90%) through the thiolysis of oxiranes by the thiols in aqueous basic catalytic conditions. The synthesized 1,2,4-triazole-tethered ß-hydroxy sulfides were screened against bacterial tyrosinase enzyme, and Gram-positive and Gram-negative bacterial cultures i.e., (S. aureus) Staphylococcus aureus & (E. coli) Escherichia coli. Among the synthesized derivatives, the molecules 11a (IC50 = 7.67 ± 1.00 µM), 11c (IC50 = 4.52 ± 0.09 µM), 11d (IC50 = 6.60 ± 1.25 µM), and 11f (IC50 = 5.93 ± 0.50 µM) displayed the better tyrosinase inhibitory activity in comparison to reference drugs ascorbic acid (IC50 = 11.5 ± 1.00 µM) and kojic acid (IC50 = 30.34 ± 0.75 µM). The molecule benzofuran-triazol-propan-2-ol 11c proved to be the most potent bacterial tyrosinase inhibitory agent with a minimum IC50 of 4.52 ± 0.09 µM, as compared to other synthesized counterparts and both standards (kojic acid and ascorbic acid). The compound diphenyl-triazol-propan-2-ol 11a and benzofuran-triazole-propan-2-ol 11c showed comparable anti-bacterial chemotherapeutic efficacy with minimum inhibitory concentrations (MIC = 2.0 ± 2.25 mg mL-1 and 2.5 ± 0.00 mg mL-1, respectively) against S. aureus bacterial strain in comparison with standard antibiotic penicillin (MIC = 2.2 ± 1.15 mg mL-1). Furthermore, among the synthesized derivatives, only compound 11c demonstrated better anti-bacterial activity (MIC = 10 ± 0.40 mg mL-1) against E. coli, which was slightly less than the standard antibiotic i.e., penicillin (MIC = 2.4 ± 1.00 mg mL-1). The compound 11c demonstrated a better binding score (-7.08 kcal mol-1) than ascorbic acid (-5.59 kcal mol-1) and kojic acid (-5.78 kcal mol-1). Molecular docking studies also validate the in vitro anti-tyrosinase assay results; therefore, the molecule 11c can be the lead bacterial tyrosinase inhibitor as well as the antibacterial agent against both types of bacterial strains after suitable structural modifications.

A Comprehensive Review on Benzofuran Synthesis Featuring Innovative and Catalytic Strategies.

Benzofurans have intrigued both pharmaceutical researchers and chemists owing to the medicinal usage of their derivatives against copious disease-causing agents (i.e., bacteria, viruses, and tumors). These heterocyclic scaffolds are pervasively encountered in a number of natural products and drugs. The ever-increasing utilization of benzofuran derivatives as pharmaceutical agents persuaded the chemists to devise novel and facile methodological approaches to assemble the biologically potent benzofuran nucleus. This review summarizes the current developments regarding the innovative synthetic routes and catalytic strategies to procure the synthesis of benzofuran heterocycles with their corresponding mechanistic details, reported by several research groups during 2021-2023.

Genome-wide analysis of NPR1-like genes in citrus species and expression analysis in response to citrus canker (Xanthomonas axonopodis pv. citri).

Citrus fruits, revered for their nutritional value, face significant threats from diseases like citrus canker, particularly impacting global citrus cultivation, notably in Pakistan. This study delves into the critical role of NPR1-like genes, the true receptors for salicylic acid (SA), in the defense mechanisms of citrus against Xanthomonas axonopodis pv. citri (Xcc). By conducting a comprehensive genome-wide analysis and phylogenetic study, the evolutionary dynamics of Citrus limon genes across diverse citrus cultivars are elucidated. Structural predictions unveil conserved domains, such as the BTB domain and ankyrin repeat domains, crucial for the defense mechanism. Motif analysis reveals essential conserved patterns, while cis-regulatory elements indicate their involvement in transcription, growth, response to phytohormones, and stress. The predominantly cytoplasmic and nuclear localization of NPR1-like genes underscores their pivotal role in conferring resistance to various citrus species. Analysis of the Ks/Ka ratio indicates a purifying selection of NPR1-like genes, emphasizing their importance in different species. Synteny and chromosomal mapping provide insights into duplication events and orthologous links among citrus species. Notably, Xac infection stimulates the expression of NPR1-like genes, revealing their responsiveness to pathogenic challenges. Interestingly, qRT-PCR profiling post-Xac infection reveals cultivar-specific alterations in expression within susceptible and resistant citrus varieties. Beyond genetic factors, physiological parameters like peroxidase, total soluble protein, and secondary metabolites respond to SA-dependent PR genes, influencing plant characteristics. Examining the impact of defense genes (NPR1) and plant characteristics on disease resistance in citrus, this study marks the inaugural investigation into the correlation between NPR1-associated genes and various plant traits in both susceptible and resistant citrus varieties to citrus bacterial canker.

Exploring theophylline-1,2,4-triazole tethered N-phenylacetamide derivatives as antimicrobial agents: unraveling mechanisms via structure-activity relationship, in vitro validation, and in silico insights.

Theophylline, a nitrogen-containing heterocycle, serves as a promising focal point for medicinal researchers aiming to create derivatives with diverse pharmacological applications. In this work, we present an improved synthetic method for a range of theophylline-1,2,4-triazole-S-linked N-phenyl acetamides (4a‒g) utilizing ultrasound-assisted synthetic approach. The objective was to assess the effectiveness of synthesized theophylline-1,2,4-triazoles (4a‒g) as inhibitors of HCV serine protease and as antibacterial agents against B. subtilis QB-928 and E. coli AB-274. Theophylline-1,2,4-triazoles were obtained in good to excellent yields (69%-95%) in a shorter time than conventional approach. 4-Chlorophenyl moiety containing theophylline-1,2,4-triazole 4c displayed significantly higher inhibitory activity against HCV serine protease enzyme (IC50 = 0.015 ± 0.25 mg) in comparison to ribavirin (IC50 = 0.165 ± 0.053 mg), but showed excellent binding affinity (-7.55 kcal/mol) with the active site of serine protease, better than compound 4c (-6.90 kcal/mol) as well as indole-based control compound 5 (-7.42 kcal/mol). In terms of percentage inhibition of serine protease, 2-chlorophenyl compound 4b showed the maximum percentage inhibition (86%), more than that of the 3,4-dichlorophenyl compound 4c (76%) and ribavirin (81%). 3,4-Dimethylphenyl-based theophylline-1,2,4-triazole 4g showed the lowest minimum inhibitory concentration (MIC = 0.28 ± 0.50 µg/mL) against the B. subtilis bacterial strain as compared to the standard drug penicillin (MIC = 1 ± 1.50 µg/mL). The other 4-methylphenyl theophylline-1,2,4-triazole 4e (MIC = 0.20 ± 0.08 µg/mL) displayed the most potent antibacterial potential against E. coli in comparison to the standard drug penicillin (MIC = 2.4 ± 1.00 µg/mL). Molecular docking studies further helped in an extensive understanding of all of the interactions between compounds and the enzyme active site, and DFT studies were also employed to gain insights into the molecular structure of the synthesized compounds. The results indicated that theophylline-linked triazole derivatives 4b and 4c showed promise as leading contenders in the fight against the HCV virus. Moreover, compounds 4e and 4g demonstrated potential as effective chemotherapeutic agents against E. coli and B. subtilis, respectively. To substantiate these findings, additional in vivo studies and clinical trials are imperative, laying the groundwork for their integration into future drug design and development.

Genetics aspect of vitamin C (Ascorbic Acid) biosynthesis and signaling pathways in fruits and vegetables crops.

Vitamin C, also known as ascorbic acid, is an essential nutrient that plays a critical role in many physiological processes in plants and animals. In humans, vitamin C is an antioxidant, reducing agent, and cofactor in diverse chemical processes. The established role of vitamin C as an antioxidant in plants is well recognized. It neutralizes reactive oxygen species (ROS) that can cause damage to cells. Also, it plays an important role in recycling other antioxidants, such as vitamin E, which helps maintain the overall balance of the plant's antioxidant system. However, unlike plants, humans cannot synthesize ascorbic acid or vitamin C in their bodies due to the absence of an enzyme called gulonolactone oxidase. This is why humans need to obtain vitamin C through their diet. Different fruits and vegetables contain varying levels of vitamin C. The biosynthesis of vitamin C in plants occurs primarily in the chloroplasts and the endoplasmic reticulum (ER). The biosynthesis of vitamin C is a complex process regulated by various factors such as light, temperature, and plant hormones. Recent research has identified several key genes that regulate vitamin C biosynthesis, including the GLDH and GLDH genes. The expression of these genes is known to be regulated by various factors such as light, temperature, and plant hormones. Recent studies highlight vitamin C's crucial role in regulating plant stress response pathways, encompassing drought, high salinity, and oxidative stress. The key enzymes in vitamin C biosynthesis are L-galactose dehydrogenase (GLDH) and L-galactono-1, 4-lactone dehydrogenase (GLDH). Genetic studies reveal key genes like GLDH and GLDH in Vitamin C biosynthesis, offering potential for crop improvement. Genetic variations influence nutritional content through their impact on vitamin C levels. Investigating the roles of genes in stress responses provides insights for developing resilient techniques in crop growth. Some fruits and vegetables, such as oranges, lemons, and grapefruits, along with strawberries and kiwi, are rich in vitamin C. Guava. Papaya provides a boost of vitamin C and dietary fiber. At the same time, red and yellow bell peppers, broccoli, pineapple, mangoes, and kale are additional sources of this essential nutrient, promoting overall health. In this review, we will discuss a brief history of Vitamin C and its signaling and biosynthesis pathway and summarize the regulation of its content in various fruits and vegetables.