Insights into the alleviation of cadmium toxicity in rice by nano-zinc and Serendipita indica: Modulation of stress-responsive gene expression and antioxidant defense system activation.

The adversities of cadmium (Cd) contamination are quite distinguished among other heavy metals (HMs), and so is the efficacy of zinc (Zn) nutrition in mitigating Cd toxicity. Rice (Oryza sativa) crop, known for its ability to absorb HMs, inadvertently facilitates the bioaccumulation of Cd, posing a significant risk to both the plant itself and to humans consuming its edible parts, and damaging the environment as well. The use of nanoparticles, such as nano-zinc oxide (nZnO), to improve the nutritional quality of crops and combat the harmful effects of HMs, have gained substantial attention among scientists and farmers. While previous studies have explored the individual effects of nZnO or Serendipita indica (referred to as S.i) on Cd toxicity, the synergistic action of these two agents has not been thoroughly investigated. Therefore, the gift of nature, i.e., S. indica, was incorporated alongside nZnO (50 mg L-1) against Cd stress (15 µM L-1) and their alliance manifested as phenotypic level modifications in two rice genotypes (Heizhan43; Hz43 and Yinni801; Yi801). Antioxidant activities were enhanced, specifically peroxidase (61.5 and 122.5% in Yi801 and Hz43 roots, respectively), leading to a significant decrease in oxidative burst; moreover, Cd translocation was reduced (85% for Yi801 and 65.5% for Hz43 compared to Cd alone treatment). Microstructural study showed a decrease in number of vacuoles and starch granules with ameliorative treatments. Overall, plants treated with nZnO displayed gene expression pattern (particularly of ZIP genes), different from the ones with alone or combined S.i and Cd. Inferentially, the integration of nZnO and S.i holds great promise as an effective strategy for alleviating Cd toxicity in rice plants. By immobilizing Cd ions in the soil and promoting their detoxification, this novel approach contributes to environmental restoration and ensures food safety worldwide.

Phytochemical screening, HPLC analysis, antimicrobial and antioxidant effect of Euphorbia parviflora L. (Euphorbiaceae Juss.).

Plant extracts are actively being used worldwide due to the presence of biologically active constituents helping in the preservation of food, and to aid against various diseases owing to their antimicrobial and antioxidant potential. The present research work was carried out to investigate the phytochemical constituents, antimicrobial activity, and antioxidant activity of different extracted samples of Euphorbia parviflora. Anti-microbial studies were carried out by Agar well diffusion while the DPPH method was employed for investigating anti-oxidant activity. Three samples from methanol, chloroform, and ethyl acetate extract were tested against five different bacterial strains comprising two species from Gram-negative bacteria i.e., Staphylococcus aureus and Bacillus subtilis and three species from Gram-positive bacteria i.e. Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumonia along two fungal strains i.e. Candida albicans and Aspergillus niger. The results of the qualitative phytochemical analysis showed that methanolic, chloroformic, and ethylacetate extract of Euphorbia parviflora consist of alkaloids, reducing sugars, flavonoids, terpenoids, tannins, and saponins. The total phenol and flavonoid content of E. parviflora showed that the methanolic extract of E. parviflora had a significantly higher total phenolic content (53.73 ± 0.30 mg of GAE/g) and flavonoid content (44.62 ± 0.38 mg of than other extracts. The content of total phenolic and flavonoids was more in methanolic extract as compared to other extracts of E. prolifera. The HPLC analysis showed that in the chloroform extract of E. parviflora Cinnamic acid (4.32 ± 2.89 mg/g) was dominant, in methanol extract quercetin (3.42 ± 2.89 mg/g) was dominant and in ethyl acetate extract of E. parviflora catechin (4.44 ± 2.89 mg/g) was found dominant. The antimicrobial activity revealed that amongst all the extracts the highest antibacterial activity was shown by methanolic extract against B. subtilis and Staphylococcus aureus as compared to the other extracts. The antioxidant activity revealed that methanolic extract of E. parviflora demonstrated higher antioxidant activity (82.42 ± 0.02) followed by chloroform extract (76.48 ± 0.08) at 150 µg/mL. The aim of this study was primarily to evaluate the potential of this plant as a reliable source of antimicrobials and antioxidants that may be used for the treatment of various infectious diseases in the future. The study provides evidence that this plant can act as a reliable source of antimicrobial and antioxidant agents and might be used against several infectious diseases.

Pronuclear transfer rescues poor embryo development of in vitro-grown secondary mouse follicles.

STUDY QUESTION: Is pronuclear transfer (PNT) capable of restoring embryo developmental arrest caused by cytoplasmic inferiority of in vitro-grown (IVG) mouse oocytes? SUMMARY ANSWER: PNT to in vivo matured cytoplasm significantly improved embryo development of IVG mouse oocytes, leading to living, fertile offspring. WHAT IS KNOWN ALREADY: In vitro follicle culture has been considered as a fertility preservation option for cancer patients. Studies describing the culture of human follicles remain scarce, owing to low availability of tissue. Mouse models have extensively been used to study and optimize follicle culture. Although important achievements have been accomplished, including the production of healthy offspring in mice, IVG oocytes are of inferior quality when compared to in vivo-grown oocytes, likely because of cytoplasmic incompetence. STUDY DESIGN SIZE DURATION: The study was carried out from September 2020 to February 2022. In total, 120 15-day-old B6D2 mice were used to perform secondary follicle culture and assess the quality of IVG oocytes. In vivo-grown control oocytes were obtained from 85 8- to 12-week-old B6D2 mice, following ovarian stimulation. For sperm collection, four B6D2 males between 10 and 14 weeks old were used. For embryo transfer, 14 8- to 12-week-old CD1 females served as surrogate mothers and 10 CD1 vasectomized males 10-24 weeks old were used to generate pseudo-pregnant females. Finally, for mating, four B6D2 female mice aged 8-10 weeks and two B6D2 male mice aged 10 weeks old were used to confirm the fertility of nuclear transfer (NT)-derived pups. PARTICIPANTS/MATERIALS SETTING METHODS: Secondary follicles from 15-day-old B6D2 mice were isolated from the ovaries and cultured for 9 days, before a maturation stimulus was given. Following 16-18 h of maturation, oocytes were collected and evaluated on maturation rate, oocyte diameter, activation rate, spindle morphology, calcium-releasing ability, and mitochondrial membrane potential. For every experiment, in vivo-grown oocytes were used as a control for comparison. When cytoplasmic immaturity and poor embryo development were confirmed in IVG oocytes, PNT was performed. For this, the pronuclei from IVG oocytes, created following parthenogenetic activation and IVF, were transferred to the cytoplasm of fertilized, in vivo-grown oocytes. Genetic analysis and embryo transfer of the generated embryos were implemented to confirm the safety of the technique. MAIN RESULTS AND THE ROLE OF CHANCE: Following 9 days of follicle culture, 703 oocytes were collected, of which 76% showed maturation to the metaphase II stage. Oocyte diameters were significantly lower in IVG oocytes, measuring 67.4 µm versus 73.1 µm in controls (P < 0.001). Spindle morphology did not differ significantly between IVG and control oocytes, but calcium-releasing ability was compromised in the IVG group. An average calcium release of 1.62 arbitrary units was observed in IVG oocytes, significantly lower than 5.74 in control oocytes (P < 0.001). Finally, mitochondrial membrane potential was inferior in IVG compared to the control group, reaching an average value of 0.95 versus 2.27 (P < 0.001). Developmental potential of IVG oocytes was assessed following parthenogenetic activation with strontium chloride (SrCl2). Only 59.4% of IVG oocytes cleaved to two cells and 36.3% reached the blastocyst stage, significantly lower than 89.5% and 88.2% in control oocytes, respectively (P < 0.001 and 0.001). Both PNT and spindle transfer (ST) were explored in pilot experiments with parthenogenetically activated oocytes, as a means to overcome poor embryo development. After the added value of NT was confirmed, we continued with the generation of biparental embryos by PNT. For this purpose, IVG and control oocytes first underwent IVF. Only 15.5% of IVG oocytes were normally fertilized, in contrast to 45.5% in controls (P < 0.001), with resulting failure of blastocyst formation in the IVG group (0 versus 86.2%, P < 0.001). When the pronuclei of IVG zygotes were transferred to the cytoplasm of control zygotes, the blastocyst rate was restored to 86.9%, a similar level as the control. Genetic analysis of PNT embryos revealed a normal chromosomal profile, to a rate of 80%. Finally, the generation of living, fertile offspring from PNT was possible following embryo transfer to surrogate mothers. LARGE-SCALE DATA: N/A. LIMITATIONS REASONS FOR CAUTION: Genetic profiles of analysed embryos from PNT originate from groups that are too small to draw concrete conclusions, whilst ST, which would be the preferred NT approach, could not be used for the generation of biparental embryos owing to technical limitations. Even though promising, the use of PNT should be considered as experimental. Furthermore, results were acquired in a mouse model, so validation of the technique in human IVG oocytes needs to be performed to evaluate the clinical relevance of the technology. The genetic profiles from IVG oocytes, which would be the ultimate characterization for chromosomal abnormalities, were not analysed owing to limitations in the reliable analysis of single cells. WIDER IMPLICATIONS OF THE FINDINGS: PNT has the ability to overcome the poor cytoplasmic quality of IVG mouse oocytes. Considering the low maturation efficiency of human IVG oocytes and potential detrimental effects following long-term in vitro culture, NT could be applied to rescue embryo development and could lead to an increased availability of good quality embryos for transfer. STUDY FUNDING/COMPETING INTERESTS: A.C. is a holder of FWO (Fonds voor Wetenschappelijk Onderzoek) grants (1S80220N and 1S80222N). B.H. and A.V.S. have been awarded with a special BOF (Bijzonder Onderzoeksfonds), GOA (Geconcerteerde onderzoeksacties) 2018000504 (GOA030-18 BOF) funding. B.H. has been receiving unrestricted educational funding from Ferring Pharmaceuticals (Aalst, Belgium). The authors declare that they have no conflict of interest.

HPLC analysis, genotoxic and antioxidant potential of Achillea millefolium L. and Chaerophyllum villosum Wall ex. Dc.

BACKGROUND: Methanolic and chloroformic extract of Achillea millefolium and Chaerophyllum villosum were evaluated for HPLC analysis, genotoxic and antioxidant potential. MATERIALS AND METHODS: Genotoxic activity was carried out on human blood lymphocytes via comet assay and antioxidant activity was studied through DPPH method. RESULTS: The genotoxic potential of A. millefolium and C. villosum's methanolic and chloroformic extract was analysed using comet assay technique. Comet shaped human lymphocytes cells were observed when treated with different concentrations (50 mg/mL, 75 mg/mL, 100 mg/mL) of methanolic and chloroformic extract of both plants. Reading was taken on the basis of damaged DNA head and tail length. Greater the length of tail as compared to head, greater will be the damage and vice versa. Total comet score was obtained from A. millefolium subjected to different concentrations. After a time interval of 24 h both the extract showed dose dependant genoprotection with maximum genoprotectivity at 98.7 ± 12.7 and 116 ± 5.3 at 50 mg/100 mL for methanolic and chloroformic extract respectively. Similarly Total Comet score was obtained from C. villosum subjected to different concentrations of methanolic and chloroformic extract. After 24 h exhibited dose dependent genoprotection with maximum protectivity at 85.7 ± 22.0 and 101.7 ± 8.6 at 50 mg/100 mL for methanolic and chloroformic extract were determined. The antioxidant activity revealed that methanolic extract of A. millefolium showed highest antioxidant activity (84.21%) at 300 mg/ml after 90 min while the chloroformic extract of C. villosum exhibited highest (68.46%) antioxidant activity (59.69%) at 300 µg/ml after 90 min but less than the standard drug ascorbic acid (88.72%). Quantitative phytochemical screening revealed high percentage of alkaloids (27.4%), Phenols (34.5%), Flavonoids (32.4%) as compared to Tannins (12%) in methanolic extract of A.millefolium. While high percentage of alkaloids (31.4), Phenols (19.3%), Flavonoids (35.5%) as compared to Tannins (16.6%) in chloroformic extract of C. villosum. CONCLUSION: The present results showed that A. millefolium and C. villosum possess a number of important compounds and revealed genoprotective property which may be used to treat several genetic disorders such as alzeimer's disease in future (Grodzicki W, Dziendzikowska K, Antioxidants 9(3):229, 2020).

Foliar zinc spraying improves assimilative capacity of sugar beet leaves by promoting magnesium and calcium uptake and enhancing photochemical performance.

Sugar beet, a zinc-loving crop, is increasingly limited by zinc deficiency worldwide. Foliar zinc application is an effective and convenient way to supplement zinc fertilizer. However, the regulatory mechanism of foliar zinc spraying on sugar beet leaf photosynthetic characteristics remains unclear. Therefore, we investigated the effects of foliar ZnSO4·7H2O application (0, 0.1%, 0.2%, and 0.4%) on the photosynthetic performance of sugar beet leaves under controlled hydroponic conditions. The results indicated that a foliar spray of 0.2% Zn fertilizer was optimal for promoting sugar beet leaf growth. This concentration significantly reduced the leaf shape index of sugar beet, notably increasing leaf area, leaf mass ratio, and specific leaf weight. Foliar spraying of Zn (0.2%) substantially elevated the Zn content in sugar beet leaves, along with calcium (Ca) and magnesium (Mg) contents. Consequently, this led to an increase in the potential photochemical activity of PSII (Fv/Fo) (by 6.74%), net photosynthetic rate (Pn) (11.39%), apparent electron transport rate (ETR) (11.43%), actual photochemical efficiency of PSⅡ (Y (Ⅱ)) (11.46%), photochemical quenching coefficient (qP) (15.49%), and total chlorophyll content (25.17%). Ultimately, this increased sugar beet leaf dry matter weight (11.30%). In the cultivation and management of sugar beet, the application of 0.2% Zn fertilizer (2.88 mg plant-1) exhibited the potential to enhance Zn and Mg contents in sugar beet, improve photochemical properties, stimulate leaf growth, and boost light assimilation capacity. Our result suggested the foliar application of Zn might be a useful strategy for sugar beet crop management.

Association between contrast-enhanced adenosine-stress perfusion cardiac magnetic resonance imaging and invasive coronary angiogram for the detection of coronary artery disease: A retrospective analysis.

Objective: The objective of this study was to determine the diagnostic value of stress perfusion CMR for the detection of coronary artery disease. Methods: The was a retrospective cross sectional study in which 43 subjects were included from Cardiac MRI unit in the Hayatabad Medical Complex, Peshawar for study from 1st April 2020 to 30th November 2020. All the subjects who had been referred for stress perfusion CMR with suspected CAD were included in the study. Cardiac MRI both at rest and with adenosine stress perfusion was performed which was followed by invasive coronary angiography. Result: A total of 43 patients were enrolled for the detection or exclusion CAD who underwent stress perfusion CMRI and invasive coronary artery angiography. The study revealed strong and statistically significant association between positive stress perfusion CMR and positive coronary angiogram vs negative stress perfusion CMR and negative coronary angiogram (p= value 0.0001). Conclusions: Stress perfusion CMRI can be considered as a first line, relatively safe, noninvasive test with significant accuracy to diagnose coronary artery disease in patients with suspected CAD without subjecting these patients to invasive coronary angiogram.

Cancer Chemoresistance; Recent Challenges and Future Considerations.

Cancer remains one of the serious health hazards and major causes of human mortality across the world. Despite the development of many typical antineoplastic drugs and introduction of novel targeted agents, chemoresistance constitutes a major challenge in the effective therapeutic management of cancer. Drug inactivation, efflux of anticancer agents, modification of target sites, enhanced repair of DNA damage, apoptosis failure and induction of epithelial-mesenchymal transition are the principal mechanisms of cancer chemoresistance. Moreover, epigenetics, cell signaling, tumor heterogeneity, stem cells, microRNAs, endoplasmic reticulum, tumor microenvironment and exosomes have also been implicated in the multifaceted phenomenon of anticancer drug resistance. The tendency of resistance is either intrinsically possessed or subsequently acquired by cancerous cells. From clinical oncology standpoint, therapeutic failure and tumor progression are the most probable consequences of cancer chemoresistance. Combination therapy can help to overcome the issue of drug resistance, and therefore, the development of such treatment regimens is recommended for counteracting the emergence and dissemination of cancer chemoresistance. This chapter outlines the current knowledge on underlying mechanisms, contributory biological factors and likely consequences of cancer chemoresistance. Besides, prognostic biomarkers, diagnostic methods and potential approaches to overcome the emergence of antineoplastic drug resistance have also been described.

Phosphorus and Serendipita indica synergism augments arsenic stress tolerance in rice by regulating secondary metabolism related enzymatic activity and root metabolic patterns.

The multifarious problems created by arsenic (As), for collective environment and human health, serve a cogent case for searching integrative agricultural approaches to attain food security. Rice (Oryza sativa L.) acts as a sponge for heavy metal(loid)s accretion, specifically As, due to anaerobic flooded growth conditions facilitating its uptake. Acclaimed for their positive impact on plant growth, development and phosphorus (P) nutrition, 'mycorrhizas' are able to promote stress tolerance. Albeit, the metabolic alterations underlying Serendipita indica (S. indica; S.i) symbiosis-mediated amelioration of As stress along with nutritional management of P are still understudied. By using biochemical, RT-qPCR and LC-MS/MS based untargeted metabolomics approach, rice roots of ZZY-1 and GD-6 colonized by S. indica, which were later treated with As (10 µM) and P (50 µM), were compared with non-colonized roots under the same treatments with a set of control plants. The responses of secondary metabolism related enzymes, especially polyphenol oxidase (PPO) activities in the foliage of ZZY-1 and GD-6 were enhanced 8.5 and 12-fold, respectively, compared to their respective control counterparts. The current study identified 360 cationic and 287 anionic metabolites in rice roots, and the commonly enriched pathway annotated by Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was biosynthesis of phenylalanine, tyrosine and tryptophan, which validated the results of biochemical and gene expression analyses associated with secondary metabolic enzymes. Particularly under As+S.i+P comparison, both genotypes exhibited an upregulation of key detoxification and defense related metabolites, including fumaric acid, L-malic acid, choline, 3,4-dihydroxybenzoic acid, to name a few. The results of this study provided the novel insights into the promising role of exogenous P and S. indica in alleviating As stress.

Lipid Lowering in "Very High Risk" Patients Undergoing Coronary Artery Bypass Surgery and Its Projected Reduction in Risk for Recurrent Vascular Events: A Monte Carlo Stepwise Simulation Approach.

ABSTRACT: 2018 AHA guidelines provide criteria to identify patients at very high risk (VHR) for adverse vascular events and recommend an low density lipoprotein-C (LDL-C) level <1.8 mmol/L. Data regarding the 10-year risk for adverse vascular events in coronary artery bypass grafting (CABG) patients at VHR and the need for nonstatin therapies in the VHR cohort are limited. We queried a national cohort of CABG patients to answer these questions. The projected reduction of LDL-C from stepwise escalation of lipid-lowering therapy (LLT) was simulated; Monte Carlo methods were used to account for patient-level heterogeneity in treatment effects. Data on preoperative statin therapy and LDL-C levels were obtained. In the first scenario, all eligible patients not at target LDL-C received high-intensity statins, followed by ezetimibe and then alirocumab; alternatively, bempedoic acid was also used. The 10-year risk for an adverse vascular event was estimated using a validated risk score. Potential risk reduction was estimated after simulating maximal LLT. Before CABG, 8948 of 27,443 patients (median LDL-C 85 mg/dL) were at VHR. In the whole cohort, 31% were receiving high-intensity statins. With stepwise LLT escalation, the proportion of patients at target were 60%, 78%, 86%, and 97% after high-intensity statins, ezetimibe, bempedoic acid, and alirocumab, respectively. The projected 10-year risk to suffer a vascular event reduced by 4.6%. A large proportion of CABG patients who are at VHR for vascular events fail to meet 2018 AHA LDL-C targets. A stepwise approach, particularly with the use of bempedoic acid, can significantly reduce the need for more expensive proprotein convertase subtilisin kexin 9 inhibitors.

Association of VEGF Gene Polymorphism (rs699947) with Glaucoma and In-Silico Study of Antiglaucoma Bioactive Compounds.

Vascular endothelial growth factor (VEGF) is a useful expert in various maturation and other problems such as glaucoma. VEGF gene is located on the short arm of chromosome 6. It has 8 exons and 6 introns. This investigation was planned to check out the relationship of VEGF polymorphism with glaucoma patients and to cause in silico limitation of carbonic anhydrase to diminish the intraocular strain of watery humor to fix glaucoma. In this examination, a large portion of the glaucoma patients (n = 70) were males (66%) when contrasted with females (34%) as the p-value was 0.025 which showed critical outcomes and subsequently demonstrated that the sickness was more predominant in males. Glaucoma for the most part influences people between the ages of 50-60 years followed by the age bunch between 30 and 40 years. Around 36 (51.4%) were experiencing watering in the eyes, 28 (40%) had photophobia, 9 (12.8%) had a hazy cornea, 6 (8.6%) had expanded eye globe, and 15 patients (21.3%) had rosy eyes. Factual tests showed that VEGF quality SNP rs699947 had no huge relationship with glaucoma (POAG). In this study, various carbonic anhydrase inhibitor phytochemicals and synthetic chemicals were screened in silico which may be used as antiglaucoma drugs.

Stress signaling convergence and nutrient crosstalk determine zinc-mediated amelioration against cadmium toxicity in rice.

Consumption of rice (Oryza sativa L.) is one of the major pathways for heavy metal bioaccumulation in humans over time. Understanding the molecular responses of rice to heavy metal contamination in agriculture is useful for eco-toxicological assessment of cadmium (Cd) and its interaction with zinc (Zn). In certain crops, the impacts of Cd stress or Zn nutrition on the biophysical chemistry and gene expression have been widely investigated, but their molecular interactions at transcriptomic level, particularly in rice roots, are still elusive. Here, hydroponic investigations were carried out with two rice genotypes (Yinni-801 and Heizhan-43), varying in Cd contents in plant tissues to determine their transcriptomic responses upon Cd15 (15 µM) and Cd15+Zn50 (50 µM) treatments. High throughput RNA-sequencing analysis confirmed that 496 and 2407 DEGs were significantly affected by Cd15 and Cd15+Zn50, respectively, among which 1016 DEGs were commonly induced in both genotypes. Multitude of DEGs fell under the category of protein kinases, such as calmodulin (CaM) and calcineurin B-like protein-interacting protein kinases (CBL), indicating a dynamic shift in hormonal signal transduction and Ca2+ involvement with the onset of treatments. Both genotypes expressed a mutual regulation of transcription factors (TFs) such as WRKY, MYB, NAM, AP2, bHLH and ZFP families under both treatments, whereas genes econding ABC transporters (ABCs), high affinity K+ transporters (HAKs) and Glutathione-S-transferases (GSTs), were highly up-regulated under Cd15+Zn50 in both genotypes. Zinc addition triggered more signaling cascades and detoxification related genes in regulation of immunity along with the suppression of Cd-induced DEGs and restriction of Cd uptake. Conclusively, the effective integration of breeding techniques with candidate genes identified in this study as well as economically and technologically viable methods, such as Zn nutrient management, could pave the way for selecting cultivars with promising agronomic qualities and reduced Cd for sustainable rice production.

Improvement of morpho-physiological, ultrastructural and nutritional profiles in wheat seedlings through astaxanthin nanoparticles alleviating the cadmium toxicity.

Heavy metal accumulation in arable lands and water bodies has become one of the serious global issues among multitude of food security challenges. In particular, cadmium (Cd) concentration has been increasing substantially in the environment that negatively affects the growth and yield of important agricultural crops, especially wheat (Triticum aestivum L.). No doubt, nanotechnology is a revolutionary science but the comprehension of nanoparticle-plants interaction and its potential alleviatory role against metal stress is still elusive. Here, we investigated the mechanistic role of astaxanthin nanoparticles (AstNPs) in Cd stress amelioration and their interaction with wheat under Cd-spiked conditions. The AstNPs fabrication was confirmed through ultraviolet visible spectroscopy, where the particles showed characteristic peak at 423 nm. However, Fourier transform infrared, X-ray diffraction, scanning electron microscopy and transmission electron microscopy analyses confirmed the presence of stabilized spherical-shaped nanocrystals of AstNPs within the size range of 12.03-30.37 nm. The hydroponic application of AstNPs (100 mg L-1) to Cd-affected wheat plants increased shoot height (59%), shoot dry weight (31%), nitrogen concentration (42%), and phosphorus concentration (26%) as compared to non-treated Cd affected seedlings. Moreover, AstNPs-treated plants showed reduction in acropetal Cd translocation (29%) in contrast to plants treated with Cd only. Under Cd-spiked conditions, AstNPs-treated plants displayed an improved nutrient profile (P, N, K+ and Ca2+) with a relative decrease in Na+ content in comparison with non-treated plants. Interestingly, it was found that AstNPs restricted the translocation of Cd to aerial plant parts by negatively regulating Cd transporter genes (TaHMA2 and TaHMA3), and relieved plants from oxidative burst by activating antioxidant machinery via triggering expressions of TaSOD and TaPOD genes. Consequently, it was observed that the application of AstNPs helped in maintaining the nutrient acquisition and ionic homeostasis in Cd-affected wheat plants, which subsequently improved the physiochemical profiles of plants under Cd-stress. This study suggests that AstNPs plausibly serve as stress stabilizers for plants under heavy metal-polluted environment.

Phyllospheric Microbial Composition and Diversity of the Tobacco Leaves Infected by Didymella segeticola.

A Myriad of biotic and abiotic factors inevitably affects the growth and production of tobacco (Nicotiana tabacum L.), which is a model crop and sought-after worldwide for its foliage. Among the various impacts the level of disease severity poses on plants, the influence on the dynamics of phyllospheric microbial diversity is of utmost importance. In China, recurring reports of a phyto-pathogen, Didymella segeticola, a causal agent of tobacco leaf spot, accentuate the need for its in-depth investigation. Here, a high-throughput sequencing technique, IonS5TMXL was employed to analyze tobacco leaves infected by D. segeticola at different disease severity levels, ranging from T1G (least disease index) to T4G (highest disease index), in an attempt to explore the composition and diversity of phyllospheric microbiota. In all healthy and diseased tobacco leaves, the most dominant fungal phylum was Ascomycota with a high prevalence of genus Didymella, followed by Boeremia, Meyerozyma and Alternaria, whereas in the case of bacterial phyla, Proteobacteria was prominent with Pseudomonas being a predominant genus, followed by Pantoea. The relative abundance of fungi, i.e., Didymella and Boeremia (Ascomycota) and bacteria, i.e., Pseudomonas and Pantoea (Proteobacteria) were higher in diseased groups compared to healthy groups. Healthy tissues exhibited relatively rich and diverse fungal communities in contrast with diseased groups. The infection of D. segeticola had a complex and significant effect on fungal as well as bacterial alpha diversity. FUNGuild analysis indicated that the relative abundance of pathotrophs and saprotrophs in diseased tissues proportionally increased with disease severity. PICRUSt analysis of diseased tissues indicated that the relative abundance of bacterial cell motility and membrane transport-related gene sequences elevated with an increase in disease severity from T1G to T3G and then tended to decrease at T4G. Conclusively, the current study shows the typical characteristics of the tobacco leaf microbiome and provides insights into the distinct microbiome shifts on tobacco leaves infected by D. segeticola.

Comparison of Kirschner Wire Versus Screw Fixation in 1,2 Inter-Compartment Supra Retinaculum Artery Pedicle-Vascularized Bone Graft for Scaphoid Fractures.

INTRODUCTION: The purpose of the study was to compare the outcomes of the Kirschner wires (k-wires) versus screw fixation in 1,2 inter-compartment supra retinaculum artery (ICSRA) pedicle-vascularized bone graft for scaphoid non-union treatment. METHOD: A retrospective study that included all patients with scaphoid nonunion underwent pedicle-vascularized bone graft and fixated with either k-wire or screw from 2010 through 2019 in the Department of Plastic and Reconstructive Surgery at Liaquat National Hospital and Medical College. Clinical outcomes were compared between k-wire and screw fixation, which were measured in terms of union rate, time of healing, pre and post-operative Disabilities of the Arm, Shoulder, and Hand (DASH) scores, range of motion at wrist, and grip strength of hand. RESULTS: A total of 33 patients were included in the study. All of them had scaphoid non-union and were treated with 1,2 ICSRA pedicle-vascularized bone graft; 20 patients had a fixation with k-wire and 13 with a screw. Radiological healing was achieved in 18 patients with k-wire and 11 patients with screw fixation, with healing rates of 90% and 84.6%, respectively. There was a significant decrease in DASH score postoperatively in both groups. Although no significant difference between the outcomes of both groups whether on union rate, DASH score, or range of motion at the wrist. CONCLUSION: Hence, there is no significant difference in outcome between k-wire and screw fixation methods. We concluded k-wire as a more acceptable option with vascularized bone graft fixation as less technically demanding and low cost as compared to screw fixation.

Vincristine induced neurotoxicity in children who underwent chemotherapy for acute lymphoblastic leukemia and Wilms tumor.

BACKGROUND & OBJECTIVES: Vincristine has been used as chemotherapeutic agent for many decades. It implements its function by inhibiting the duplication of tumor cells by destroying the DNA. However, like all other drugs, its administration is not without any side effects. The most important of these are being the neurotoxic side effects. This study evaluated the degree of neurotoxicity induced by vincristine in children who underwent chemotherapy for acute lymphoblastic leukemia and Wilms tumor. METHODS: A quasi experimental study was conducted at Children Hospital & the Institute of Child Health, Multan from January 2020 to October 2020 after taking informed written consent. In this study, 150 children of age group 1 - 12 years with pathological confirmation of acute lymphoblastic leukemia and Wilms tumor who had undergone a chemotherapy protocol including at least four consecutive weekly Vincristine injections were included, using probability consecutive sampling technique. Neurological examination was conducted on them on weekly basis. RESULTS: There were 150 patients,90(60%) males and 60(40%) females with mean age of (5.5±2.2). Diminished patellar and Achilles tendon reflexes were seen in 48% and 52% of patients. Muscular weakness was seen in 60% of patients. Other side effects like hoarseness, jaw pain, constipation and petosis were observed in 10%, 8%,40% and 10% of patients respectively. Frequency of side effects was equally observed in both sexes and it was more among age group older than five years (p= 0.01). CONCLUSION: Vincristine regimen produces some neurotoxic side effects in children but nearly all of these are of mild to moderate in nature.

Physio-ultrastructural footprints and iTRAQ-based proteomic approach unravel the role of Piriformospora indica-colonization in counteracting cadmium toxicity in rice.

Due to its immense capability to concentrate in rice grain and ultimately in food chain, cadmium (Cd) has become the cause of an elevated concern among agriculturists, scientists and the environmental activists. Symbiotic association of Piriformospora indica (P. indica) has been characterized as a potential aid in combating heavy metal stress in plants for sustainable crop production but our scant knowledge regarding ameliorative tendency of P. indica against Cd, specifically in rice, necessitates an in-depth investigation. This study aimed at elaborating the underlying mechanisms involved in P. indica-mediated tolerance against Cd stress in two rice genotypes, IR8 and ZX1H, varying in Cd accumulation pattern. Either colonized or un-inoculated with P. indica, seedlings of both genotypes were subjected to Cd stress. The results showed that P. indica colonization significantly supported plant biomass, photosynthetic attributes and chlorophyll contents in Cd stressed plants. P. indica colonization sustained chloroplast integrity and reduced Cd translocation (46% and 64%), significantly lowering malondialdehyde (MDA) content (11.3% and 50.4%) compared to uninoculated roots under Cd stress in IR8 and ZX1H, respectively. A genotypic difference was evident when a 2-fold enhancement in root peroxidase (POD) activity was recorded in P. indica colonized IR8 plants as compared to ZX1H. The root proteomic analysis was performed using isobaric tags for relative and absolute quantification (iTRAQ) and the results showed that P. indica alleviates Cd stress in rice via down-regulation of key glycolysis cycle enzymes in a bid to reduce energy consumption by the plants and possibly re-directing it to Cd defense response pathways; and up-regulation of glutamine synthetase, a key enzyme in the L-Arg-dependent pathway for nitric oxide (NO) production, which acts as a stress signaling molecule, thus conferring tolerance by reduction of NO-mediated modification of essential proteins in response to Cd stress. Conclusively, both the tested genotypes benefited from P. indica symbiosis at varying levels by an enhanced detoxification capacity and signaling efficiency in response to stress. Hence, a step forward towards the employment of an environmentally sound and self-renewing approach holding the hope for a healthy future.

Modulation of Key Physio-Biochemical and Ultrastructural Attributes after Synergistic Application of Zinc and Silicon on Rice under Cadmium Stress.

Excessive industrialization and the usage of pesticides plague the farming soils with heavy metals, reducing the quality of arable land. Assessing phytoavailability of cadmium (Cd) from growth medium to plant system is crucial and necessitates precise and timely monitoring of Cd to ensure food safety. Zinc (Zn) and silicon (Si) have singularly demonstrated the potential to ameliorate Cd toxicity and are important for agricultural production, human health, and environment in general. However, Zn-Si interaction on Cd toxicity alleviation, their effects and underlying mechanisms are still fragmentarily understood. Seven treatments were devised besides control to evaluate the single and combined effects of Zn and Si on the physio-biochemical attributes and ultrastructural fingerprints of Cd-treated rice genotypes, i.e., Cd tolerant "Xiushui-110" and Cd sensitive "HIPJ-1". Supplementation of both Zn and Si promoted plant biomass, photosynthetic parameters, ionic balance, and improved chloroplast ultrastructure with minimized Cd uptake and malondialdehyde (MDA) content due to the activation of antioxidant enzymes in Cd stressed plants. The combined effects of 10 µM Zn and 15 µM Si on 15 µM Cd displayed a greater reduction in Cd uptake and root-leaf MDA content, while enhancing photosynthetic activity, superoxide dismutase (SOD) activity and root-leaf ultrastructure particularly in HIPJ-1, whilst Xiushui-110 had an overall higher leaf catalase (CAT) activity and a higher root length and shoot height was observed in both genotypes compared to the Cd 15 µM treatment. Alone and combined Zn and Si alleviation treatments reduced Cd translocation from the root to the stem for HIPJ-1 but not for Xiushui-110. Our results confer that Zn and Si singularly and in combination are highly effective in reducing tissue Cd content in both genotypes, the mechanism behind which could be the dilution effect of Cd due to improved biomass and competitive nature of Zn and Si, culminating in Cd toxicity alleviation. This study could open new avenues for characterizing interactive effects of simultaneously augmented nutrients in crops and provide a bench mark for crop scientists and farmers to improve Cd tolerance in rice.

RP-HPLC-based phytochemical analysis and diverse pharmacological evaluation of Quercus floribunda Lindl. ex A. Camus nuts extracts.

Quercus floribunda Lindl. ex A. Camus nuts have important folklore uses, assessed for underexplored biological potential. Nuts galls or cores and coats were utilized for the preparation of extracts using 14 solvent systems. Antioxidant, antimicrobial, cytotoxic and enzyme inhibition assays along with phytochemical profiling was carried out. Distilled water cores extract demonstrated maximum percent yield, phenolics content and total antioxidant capacity. Methanol: ethyl acetate cores extract showed maximum flavonoids content, total reducing power and protein kinase inhibition. Highest percentage radical scavenging and brine shrimp lethality was revealed by acetone: distilled water cores extract. Ethyl acetate cores extract indicated maximum α-amylase inhibition. Methanol: water coats extract showed substantial leishmanial growth inhibition. n-Hexane and chloroform coats extracts showed maximum cytotoxicity against HepG2 and THP-1 cell lines, respectively. Polyphenols quantified through RP-HPLC analysis were quercetin, pyrocatechol, gallic acid, catechin and chlorogenic acid ranging from 0.003 ± 0.001 to 1.785 ± 0.5 µg/mg extract.

In vitro antimutagenic, cytotoxic and anticancer potential of Fagonia indica phytochemicals.

Cancer is one of the most diagnosed and life threatening disease throughout the world. Nevertheless present day clinical management for cancers are surgery, radiations which are insufficient to contain the disease burden. In the past two decades, more than half of chemotherapeutic drugs developed are either directly or indirectly dependent on medicinal base phytocompounds or their derivative. The present study aims to provide the base for chemotherapeutic phytochemicals. Fagonia indica showed significant antimutagenic potential with reference to control IC50 values were calculated as 146.33±5.2µg/ml, TA100 (AZS) 105.33±4.0µg/ml, TA98 (2AA) 113.6±5.2µg/ml followed and TA98 (AZS) 112.6±4.4 in Ames test. For this reason, the antiproliferation effect of extracts on cancer cell lines was studied through resazurin fluorescence. On HepG-2 cell lines 50% cytotoxic concentration (CC50) of (FIWM) was recorded as 128.3±,2.43µg/ml. On the homo sapiens epithelial cell of lung tissue (A549), the high throughput instrumental analysis of Fagonia indica depicts maximum cytotoxic effect in 30hr. The electrical impedance displays the real-time evidence about qualitative apoptosis expressed. The impedance results were supported as palmitic acid from Fagonia indica virtually that inhibits Cyclin Dependent Kinase 2 (CDKs 2) in silico molecular docking studies. Fagonia indica extract possesses substantial antimutagenic, cytotoxic and anticancer activity which supports the potential of its phytochemicals for drug development.