Genome-wide identification and characterization of wall-associated kinases, molecular docking and polysaccharide elicitation of monoterpenoid indole alkaloids in micro-propagated Catharanthus roseus.

Wall-associated kinases (WAKs) are a unique family of proteins that are predominantly localized on the plasma membrane and simultaneously bound to the cell wall. WAKs play a pivotal role in signal transduction to regulate growth, defense, and response to environmental stimuli in plants. These kinases have been identified and characterized in various plant species, however, similar information for Catharanthus roseus is scarce. C. roseus is an evergreen ornamental plant that produces a repertoire of biologically active compounds. The plant is best characterized for the production of antineoplastic monoterpenoid indole alkaloids (MIAs) namely vinblastine and vincristine. Owing to the diverse composition of phytochemicals, C. roseus is known as a "model non-model" plant for secondary metabolite research. Genome analyses showed 37 putative CrWAK genes present in C. roseus, largely localized on the plasma membrane. Phylogenetic analysis revealed six clusters of CrWAKs. Diverse cis-acting elements, including those involved in defense responses, were identified on the promotor regions of CrWAK genes. The highest binding affinity (- 12.6 kcal/mol) was noted for CrWAK-22 against tri-galacturonic acid. Tri-galacturonic acid stimulated 2.5-fold higher production of vinblastine, sixfold upregulation of the expression of ORCA3 transcription factor, and 6.14-fold upregulation of CrWAK-22 expression. Based on these results it was concluded that the expression of CrWAK genes induced by biotic elicitors may have an important role in the production of MIAs. The current findings may serve as a basis for functional characterization and mechanistic explanation of the role of CrWAK genes in the biosynthesis of MIAs upon elicitation.

Real-time expression and in silico characterization of pea genes involved in salt and water-deficit stress.

BACKGROUND: To tolerate salt and water-deficit stress, the plant adapts to the adverse environment by regulating its metabolism and expressing certain stress-induced metabolic pathways. This research analyzed the relative expression of four pea genes (P5CR, PAL1, SOD, and POX) in three pea varieties (Climax, Green grass, and Meteor) under different levels of salt and water-deficit stress. METHODS AND RESULTS: The experiments on salt stress and water-deficit stress were carried out within greenhouse settings under controlled environment. The saturation percentage was employed to create artificial salinity conditions: Control without NaCl treatment, Treatment 1: 50 mM NaCl treatment, Treatment 2: 75 mM NaCl treatment, and Treatment 3: 100 mM NaCl treatment. Field capacity (FC) was used for the development of artificial water-deficit treatments in the pots, i.e., Treatment 1 (Control; water application 100% of FC), Treatment 2 (water application 75% of FC), and Treatment 3 (water application 50% of FC). Pea genes involved in biosynthetic pathways of proline, flavonoids, and enzymatic antioxidant enzymes including P5CR, PAL1, SOD, and POX were selected based on literature. Quantitative real-time PCR using cDNA as a template was used to analyze the gene expression. Pea genes were analyzed for phylogenetic analysis in closely related crops having similarity percent identity 80% and above. In silico characterization of selected proteins including the family classification was done by the NCBI CDD and INTERPRO online servers. Results from RT-qPCR analysis showed increased expression of P5CR, PAL1, and POX genes, while SOD gene expression decreased under both stresses. Climax exhibited superior stress tolerance with elevated expression of P5CR and PAL1, while Meteor showed better tolerance through increased POX expression. Phylogenetic analysis revealed common ancestry with other species like chickpea, red clover, mung bean, and barrel clover, suggesting the cross relationship among these plant species. Conserved domain analysis of respective proteins revealed that these proteins contain PLNO 2688, PLN02457, Cu-Zn Superoxide dismutase, and secretory peroxidase conserved domains. Furthermore, protein family classification indicated that the oxidation-reduction process is the most common chemical process involved in these stresses given to pea plant which validates the relationship of these proteins. CONCLUSIONS: Salt and water-deficit stresses trigger distinct metabolic pathways, leading to the up-regulation of specific genes and the synthesis of corresponding proteins. These findings further emphasize the conservation of stress-tolerance-related genes and proteins across various plant species. This knowledge enhances our understanding of plant adaptation to stress and offers opportunities for developing strategies to improve stress resilience in crops, thereby addressing global food security challenges.

The Potential and Concerns of Using AI in Scientific Research: ChatGPT Performance Evaluation.

BACKGROUND: Artificial intelligence (AI) has many applications in various aspects of our daily life, including health, criminal, education, civil, business, and liability law. One aspect of AI that has gained significant attention is natural language processing (NLP), which refers to the ability of computers to understand and generate human language. OBJECTIVE: This study aims to examine the potential for, and concerns of, using AI in scientific research. For this purpose, high-impact research articles were generated by analyzing the quality of reports generated by ChatGPT and assessing the application's impact on the research framework, data analysis, and the literature review. The study also explored concerns around ownership and the integrity of research when using AI-generated text. METHODS: A total of 4 articles were generated using ChatGPT, and thereafter evaluated by 23 reviewers. The researchers developed an evaluation form to assess the quality of the articles generated. Additionally, 50 abstracts were generated using ChatGPT and their quality was evaluated. The data were subjected to ANOVA and thematic analysis to analyze the qualitative data provided by the reviewers. RESULTS: When using detailed prompts and providing the context of the study, ChatGPT would generate high-quality research that could be published in high-impact journals. However, ChatGPT had a minor impact on developing the research framework and data analysis. The primary area needing improvement was the development of the literature review. Moreover, reviewers expressed concerns around ownership and the integrity of the research when using AI-generated text. Nonetheless, ChatGPT has a strong potential to increase human productivity in research and can be used in academic writing. CONCLUSIONS: AI-generated text has the potential to improve the quality of high-impact research articles. The findings of this study suggest that decision makers and researchers should focus more on the methodology part of the research, which includes research design, developing research tools, and analyzing data in depth, to draw strong theoretical and practical implications, thereby establishing a revolution in scientific research in the era of AI. The practical implications of this study can be used in different fields such as medical education to deliver materials to develop the basic competencies for both medicine students and faculty members.

Evaluating the antioxidative defense response of selected indoor plants against benzene and formaldehyde.

Volatile organic compounds (VOCs) such as formaldehyde and benzene are among the key contributors to indoor air pollution. The current situation of environmental pollution is alarming, especially indoor air pollution is becoming a challenge as affecting plants and humans. VOCs are known to adversely affect indoor plants by causing necrosis and chlorosis. In order to withstand these organic pollutants, plants are naturally equipped with an antioxidative defense system. The current research study aimed to evaluate the combined effect of formaldehyde and benzene on the antioxidative response of selected indoor C3 plants including Chlorophytum comosum, Dracaena mysore, and Ficus longifolia. After the combined application of different levels (0, 0; 2, 2; 2, 4; 4, 2; and 4, 4 ppm) of benzene and formaldehyde respectively, in an airtight glass chamber, the enzymatic and non-enzymatic antioxidants were analyzed. Analysis of total phenolics showed a significant increase (10.72 mg GAE/g) in F. longifolia; C. comosum (9.20 mg GAE/g); and D. mysore (8.74 mg GAE/g) compared to their respective controls as 3.76, 5.39, and 6.07 mg GAE/g. Total flavonoids (724 µg/g) were reported in control plants of F. longifolia which were increased to 1545.72 µg/g from 724 µg/g (in control) followed by 322.66 µg/g in D. mysore (control having 167.11 µg/g). Total carotenoid content also increased in D. mysore (0.67 mg/g) followed by C. comosum (0.63 mg/g) in response to increasing the combined dose compared to their control plants having 0.62 and 0.24 mg/g content. The highest proline content was exhibited by D. mysore (3.66 µg/g) as compared to its respective control plant (1.54 µg/g) under a 4 ppm dose of benzene and formaldehyde. A significant increase in enzymatic antioxidants including total antioxidants (87.89%), catalase (59.21 U/mg of protein), and guaiacol peroxidase (52.16 U/mg of protein) was observed in the D. mysore plant under a combined dose of benzene (2 ppm) and formaldehyde (4 ppm) with respect to their controls. Although experimental indoor plants have been reported to metabolize indoor pollutants, the current findings indicate that the combined application of benzene and formaldehyde is also affecting the physiology of indoor plants as well.

Knowledge, attitude, opinion, perspective, and agreement of Palestinian medical students on strategies/recommendations to curb plagiarism: A multicenter cross-sectional study.

Plagiarism is a common issue in written academic assignments and graduation theses. This multicenter study was conducted to assess the knowledge, attitude, opinion, perspective, and agreement of Palestinian medical students on strategies/recommendations to curb plagiarism. The study was conducted in a cross-sectional design using a questionnaire in all universities with medical education programs. The questionnaire contained 12 knowledge items, 8 attitude items, 6 opinion/perspective items, and 8 strategies/recommendations to curb plagiarism. Of the 550 invited medical students, 474 completed the study tool. Knowledge, attitude, opinion, perspective, and agreement on strategies/recommendations scores correlated positively. Higher knowledge, attitude, opinion, perspective, and agreement on strategies/recommendations scores were significantly associated with higher academic/training year, grade point average, satisfaction with academic achievement, academic writing skills, informational skills, using citation managers, receiving courses/workshops/lecturers on plagiarism, using plagiarism checking tools, and participation in a scientific paper/graduation thesis writing. Gaps in knowledge about plagiarism were identified among Palestinian medical students. Educators/trainers and other decision-makers in medical schools and higher academic institutions might use the strategies on which the students agreed to curb plagiarism.

Genome-wide identification of NAC transcription factors and regulation of monoterpenoid indole alkaloid biosynthesis in Catharanthus roseus.

NAC transcription factors (TFs) are crucial to growth and defense responses in plants. Though NACs have been characterized for their role in several plants, comprehensive information regarding their role in Catharanthus roseus, a perennial ornamental plant, is lacking. Homology modelling was employed to identify and characterize NACs in C. roseus. In-vitro propagation of C. roseus plants was carried out using cell suspension and nodal culture and were elicited with two auxin-antagonists, 5-fluoro Indole Acetic Acid (5-F-IAA) and α-(phenyl ethyl-2-oxo)-Indole-Acetic-Acid (PEO-IAA) for the enhanced production of monoterpenoid indole alkaloids (MIAs) namely catharanthine, vindoline, and vinblastine. Analyses revealed the presence of 47 putative CrNAC genes in the C. roseus genome, primarily localized in the nucleus. Phylogenetic analysis categorized these CrNACs into eight clusters, demonstrating the highest synteny with corresponding genes in Camptotheca acuminata. Additionally, at least one defense or hormone-responsive cis-acting element was identified in the promoter region of all the putative CrNACs. Of the two elicitors, 5-F-IAA was effective at 200 µM to elicit a 3.07-fold increase in catharanthine, 2.76-fold in vindoline, and 2.4-fold in vinblastine production in nodal culture. While a relatively lower increase in MIAs was recorded in suspension culture. Validation of RNA-Seq by qRT-PCR showed upregulated expression of stress-related genes (CrNAC-07 and CrNAC-24), and downregulated expression of growth-related gene (CrNAC-25) in elicited nodal culture of C. roseus. Additionally, the expression of genes involved in the biosynthesis of MIAs was significantly upregulated upon elicitation. The current study provides the first report on the role of CrNACs in regulating the biosynthesis of MIAs.

Factors Affecting Glycemic Control among Saudi Children with Type 1 Diabetes Mellitus in Aseer Region, Southwestern Saudi Arabia.

Glycemic control in children with type 1 diabetes mellitus (T1DM) is affected by many factors that may be influenced by their lives and community. To identify the factors associated with glycemic control among children with T1DM in Aseer Region, southwestern Saudi Arabia, a cross-sectional interview study was conducted between 1 July and 30 September 2021, with a representative sample of Saudi children aged between 6 months and 15 years with T1DM or their caregivers visiting the diabetes center at Aseer Region. Newly diagnosed cases (<12 months) were excluded from the study. The study included 171 T1DM pediatric patients aged between 18 months and 15 years. The glycated hemoglobin (HbA1c) level ranged between 6.10% and 15.10% (mean HbA1c = 10.39% ± 1.86%). High HbA1c levels (≥7.5%) were observed in most patients (94.7%). Only two significant factors were found: (1) use of carbohydrate counting; 81.8% of children using carbohydrate counts had high HbA1c levels, compared to 96.6% of children not using carbohydrate counts (p = 0.017), and (2) duration of the disease; 91.5% of children with disease duration of ≤3 years had high levels of HbA1c, compared to 98.7% of children with disease duration exceeding 3 years (p = 0.035). Most children with T1DM in Aseer Region had poor glycemic control. Only two factors were associated with better glycemic control: shorter disease duration and use of carbohydrate counting. Therefore, advising diabetic patients to be on a carbohydrate counting program might improve DM control.

Multiwalled carbon nanotubes functionalized bacterial cellulose as an efficient healing material for diabetic wounds.

The unique pool of features makes bacterial cellulose (BC) a robust platform to tailor its functionalities. Herein, the BC matrix was reinforced with multiwalled carbon nanotubes (MWCNT) to control infection and accelerate the healing process of diabetic wounds. The prepared BC-MWCNT composite film was characterized and antibacterial activity was assessed. Further, the in-vivo wound healing activity was performed and temporal expression of interleukin (IL-1α), tumor necrosis factor (TNF-α), vascular endothelial growth factor (VEGF) and platelets derived growth factor (PDGF) was quantitatively measured by real-time PCR. The characterization results confirmed the reinforcement of the BC matrix with MWCNT. The composite film showed antibacterial activity against all the tested strains. Moreover, the macroscopic analysis of the wound demonstrated faster closure of the diabetic wound in BC-MWCNT group (99% healing) as compared to negative control (77%) in 21 days. Histological studies further supported the results where complete reepithelization of the epidermis and healthy granulation tissue were observed in BC-MWCNT treated group. Molecular studies revealed that BC-MWCNT group showed relatively lesser expression of pro-inflammatory cytokines IL-1α and TNF-α and higher expression of VEGF than control that may have favored the faster healing. This study suggested that the tailorable properties of BC can be exploited to develop composites with potential applications in diabetic wound healing.

Identification of differentially expressed genes and pathways crosstalk analysis in Rheumatoid and Osteoarthritis using next-generation sequencing and protein-protein networks.

Osteoarthritis occurs when protective cartilage of bones worn out. Similarlty, cartilage damage occurs mainly in the pannus cartilage in rheumatoid arthritis. It is a potentially debilitating condition, affecting women two to three times more often than men. The cause and prognosis of rheumatoid and osteoarthritis are still poorly known. However, advances in the study of disease pathogenesis have encouraged the creation of new therapeutics with improved outcomes. The purpose of this study is to investigate the differentially expressed genes potentially involved in dysregulated rheumatoid arthritis (RA) and their association to other types of arthritis, including osteoarthritis (OA). Complete RNAs were isolated for RNA expression profiling using next-generation sequencing from human primary cultured normal and RA chondrocytes. From RNA sequencing results 250 differentially expressed genes were identified using bioinformatics analysis, of which 32 were found to be significantly playing role in RA pathogenesis and its associated diseases. Molecular ontologies of the identified genes showed they are connected to Innate immune response, Protein phosphorylation, Transcription initiation from RNA polymerase II promoter, Immune response, Neoplasms of bones, as well as osteorthritis, and Rheumatoid arthritis. Among the identified genes, TRAF1, TRAF2, BAMP, STX11, MEOX2, AES, REL, FHL3, PNMA1, SGTA, LZTS2, SIAH2, PNMA1, and TFCP2 were found to be highly enriched in the protein-protein interaction network. The significant cross talks were found in Hypertrophic cardiomyopathy, Small cell lung cancer, Proteasome, p53 signaling pathway, Arrhythmogenic right ventricular cardiomyopathy, Small cell lung cancer, SNARE interactions in vesicular transport, RIG-I-like receptor signaling pathway, and Hypertrophic cardiomyopathy pathways. The results offer new opportunities for target gene control in RA and OA cartilage destruction.

Fractionation of phosphorus in soils amended with poultry manure co-composted with sugarcane and cabbage wastes / Fracionamento de fósforo em solos corrigidos com esterco de aves co-compostado com resíduos de cana de açúcar e repolho

Organic waste recycling is a viable option for reducing energy usage, volume of landfills, air and water pollution, greenhouse gas emissions and preserving natural resources for future use. Composting is an easy and natural bio-degradation process that converts organic wastes into available nutrients for plants. We studied the changes in phosphorus (P) fractions in soils amended with poultry manure co-composted with sugarcane and cabbage wastes. The compost was applied to sandy clay and silt loam soils at 10 and 20 t ha-1. Soils were then incubated at room temperature for 8 weeks when mineralization was expected and analyzed for extractable P fractions. The P fractions in the soils varied in the order HCl-P (Ca+Mg-bound) > H2O-P (water soluble) > NaHCO3-P (readily plant-available P) > NaOH-P (Fe+Albound) and the fractions increased significantly as compost application rates increased and decreased as the amount of sugarcane and cabbage wastes in the compost increased. Phosphorus was less concentrated in the compost containing CW than that containing SW and was higher in sandy clay than silty loam soil. The overall results showed that composting reduced the bio-availability of P from poultry litter and would be beneficial for optimizing P fertility in soil and minimizing losses to the environment.

A reciclagem de resíduos orgânicos é uma opção viável para reduzir o uso de energia, o volume de aterros sanitários, a poluição do ar e da água, as emissões de gases de efeito estufa e a preservação dos recursos naturais para uso futuro. A compostagem é um processo fácil e natural de biodegradação que converte resíduos orgânicos em nutrientes disponíveis para plantas. Estudamos as alterações nas frações de fósforo (P) em solos alterados com esterco de aves de capoeira co-compostada com resíduos de cana de açúcar e repolho. O composto foi aplicado em solos arenosos, argilosos e limosos com 10 e 20 t ha-1. Os solos foram então incubados à temperatura ambiente durante 8 semanas quando a mineralização era esperada e analisada para as frações de P extraíveis. As frações de P nos solos variaram na ordem HCl- P (Ca + Mg-bound) > H2O-P (solúvel em água) > NaHCO3-P ( P prontamente disponível para a planta) > NaOH-P (Fe + Al-bound) e as frações aumentaram significativamente à medida que as taxas de aplicação de composto aumentaram e diminuíram à medida que aumentou a quantidade de resíduos de cana de açúcar e de repolho na compostagem. O fósforo estava menos concentrado no composto contendo CW do que o que continha SW e era mais alto em argila arenosa do que o solo limoso. Os resultados globais mostraram que a compostagem reduziu a biodisponibilidade do P vindo do lixo de aves de capoeira e seria benéfica para otimizar a fertilidade do P no solo e minimizar as perdas para o meio ambiente.

Epigenetic regulation of inflammatory cytokines and associated genes in human malignancies.

Inflammation is a multifaceted defense response of immune system against infection. Chronic inflammation has been implicated as an imminent threat for major human malignancies and is directly linked to various steps involved in tumorigenesis. Inflammatory cytokines, interleukins, interferons, transforming growth factors, chemokines, and adhesion molecules have been associated with chronic inflammation. Numerous cytokines are reported to be aberrantly regulated by different epigenetic mechanisms like DNA methylation and histone modifications in tumor tissues, contributing to pathogenesis of tumor in multiple ways. Some of these cytokines also work as epigenetic regulators of other crucial genes in tumor biology, either directly or indirectly. Such regulations are reported in lung, breast, cervical, gastric, colorectal, pancreatic, prostate, and head and neck cancers. Epigenetics of inflammatory mediators in cancer is currently subject of extensive research. These investigations may help in understanding cancer biology and to develop effective therapeutic strategies. The purpose of this paper is to have a brief view of the aberrant regulation of inflammatory cytokines in human malignancies.