Phytochemicals analysis and aflatoxin B1 detoxification potential of leaves extract of Moringa oleifera and Calotropis procera.

The present study aimed to identify the presence of certain classes of phytochemicals in the leaf extract of medicinal herbs viz. Moringa oleifera and Calotropis procera, using qualitative detection tests and explored the potential of aqueous and ethanolic extract to inhibit aflatoxin production by thin layer chromatography at 25 °C and pH (7) of different incubation times i.e. 0-, 1-, 3-, 6- and 24-h. Qualitative phytochemical analysis reported that the aqueous leave extracts of M. oleifera and C. procera contained tannins, phlobatannins, quinones, steroids, sugar, betacyanins, fatty acids, phenols, and volatile oils. Aflatoxin analysis reported that the ethanolic extract of M. oleifera was found more effective than detoxifying 100% of AFB1 after 24 h of incubation. In the case of C. procera, the aqueous extract reduced 96.5% of AFB1 and ethanolic extract reduced 96% of AFB1 after 24-h of incubation. The results revealed that natural plant products have a high potential to reduce AFB1 and could contribute to mitigation plans for AFB1. There is a need for further characterisation using techniques such as GC-MS, LC-MS, or NMR which would provide valuable information on the chemical composition of the extracts.

Proximate composition and prevalence and exposure assessment of aflatoxins intake through consumption of fresh carrot and processed marketed carrot products in South Punjab, Pakistan.

Carrot (Daucus carota L.) is a nutrient-rich vegetable that is widely cultivated and consumed in Pakistan in both raw and processed form. Data on the proximate composition and natural occurrence of aflatoxins (AFs) in carrots and marketed carrot products is lacking in Pakistan and the risk exposure of AF has not been characterised before. Thus, the current study was designed to know the frequently consumed carrot products with per capita consumption, and risk assessment of AF through these products in various regions of South Punjab Pakistan. A survey was conducted with 125 respondents and appeared that raw carrot, fresh carrot juice, gajrella and pickle are the most frequently consumed marketed carrot products with per capita consumption i.e. 62.5, 46.6, 16.2 and 14.5 gday-1, respectively. Proximate analysis revealed that carrot root and processed carrot products contained 9.65-98.2% moisture, 0.23-0.60% ash, 6.2-14.1% carbohydrates, 0.31-0.80% protein, 0.40-3.7% fat and 1.4-4.20% fibre. AF analysis revealed that 36.67% of samples were contaminated with TAF. Thirty-five (35%) percent of samples were tainted with aflatoxin B1, and 13.33% of samples were contaminated with aflatoxin B2. All the samples of carrot root, fresh carrot juice and gajrella contained TAF levels less than the maximum limit (ML) (2 ppb) assigned by the European Union (EU). However, the entire AFB1 positive samples of carrot pickle contained AFB1 levels of more than 2 ppb exceeding the ML. Furthermore, daily dietary exposure of TAFs ranged from 0.11 to 1.27 ng/kg of body weight per day which relatively exceeds the permissible limit of 1 ng/kg of body weight per day as defined by the Joint FAO/WHO Expert Committee on Food Additives. This is the first prevalence and risk assessment report of AF in marketed processed carrot products in Pakistan. These baseline data are an initial step in the effort to deal with this significant food safety issue and the establishment of legislation for AF in marketed products is needed in Pakistan.

Biosynthesized silver nanoparticles enhanced wheat resistance to Bipolaris sorokiniana.

Agronomic crops can benefit from the application of nanoscale materials in order to control phytopathogens and improve plant growth. Bipolaris sorokiniana, a soil- and seed-borne fungus, causes severe yield losses in wheat. In order to determine the physio-chemical changes in wheat under biotic stress of B. sorokiniana, the current study aimed to synthesis silver nanoparticles (AgNPs) using Allium sativum bulb extract. Herein, we applied the silver nanoparticles (AgNPs) as a foliar spray on two wheat varieties (Pakistan-2013, and NARC-2011) at the concentrations of 10, 20, 30, and 40 mg/L to suppress B. sorokiniana. Among all the applied concentrations of AgNPs, the 40 mg/L concentration demonstrated the most effective outcome in reduction of the intensity of spot blotch and improved the morphological, physiological, biochemical parameters, as well as antioxidant activity in wheat plant. Foliar application of AgNPs at 40 mg/L Pakistan-2013 and NARC-2011 wheat varieties significantly increased chlorophyll a 84.8% and 53.4%, chlorophyll b 28.9% and 84.3%, total chlorophyll content 294.3% and 241.2%, membrane stability index 7.5% and 6.1%, relative water contents 25.4% and 10.5%, proline content 320.5% and 609.9%, and soluble sugar content 120% and 259.4%, respectively, compared to control and diseased plant. This is the first study provides important insights into the role of phyto-mediated AgNPs in increasing resistant of wheat infected with B. sorokiniana. These findings offers valuable new insights that may be useful for reducing disease incidence in wheat fields.

Effects of inoculation of corn silage with Saccharomyces cerevisiae on silage fermentation characteristics, nutrient digestibility, mycoflora and aflatoxin production.

In the present study, fresh whole corn plants along with half milk kernels line were treated with live and hydrolysed yeast with different concentrations before ensiling and kept in airtight bags and then into mini silos in order to achieve anaerobic conditions for proper fermentation. The buckets were opened after different time intervals to characterise the material, quick acidification, dry matter recovery, and aerobic stability of silage respectively. Moreover, mycoflora and aflatoxin contamination were also analysed. The overall result reported that the silage quality was improved by the application of live and hydrolysed yeast. The best result was reported by the application of live yeast (T2: 10 g/kg) which significantly improved the fermentative, proximate, and digestibility parameters and reduced the mycoflora and aflatoxin contamination. Our results present promising new options for the use of natural compounds that may help to improve silage quality and reduce aflatoxin contamination.

Antifungal activity of green synthesized selenium nanoparticles and their effect on physiological, biochemical, and antioxidant defense system of mango under mango malformation disease.

Plant extract-based green synthesis of nanoparticles is an emerging class of nanotechnology that has revolutionized the entire field of biological sciences. Green synthesized nanoparticles are used as super-growth promoters and antifungal agents. In this study, selenium nanoparticles (SeNPs) were synthesized using Melia azedarach leaves extract as the main reducing and stabilizing agent and characterized by UV-visible spectroscopy, scanning electron microscopy (SEM), X-ray diffraction (XRD), energy-dispersive X-ray (EDX), and fourier transform infrared spectrometer (FTIR). The green synthesized SeNPs were exogenously applied on Mangifera indica infected with mango malformation disease. The SeNPs at a concentration of 30 µg/mL were found to be the best concentration which enhanced the physiological (chlorophyll and membrane stability index), and biochemical (proline and soluble sugar) parameters. The antioxidant defense system was also explored, and it was reported that green synthesized SeNPs significantly reduced the biotic stress by enhancing enzymatic and non-enzymatic activities. In vitro antifungal activity of SeNPs reported that 300 µg/mL concentration inhibited the Fusarium mangiferae the most. This study is considered the first biocompatible approach to evaluate the potential of green synthesized SeNPs to improve the health of mango malformation-infected plants and effective management strategy to inhibit the growth of F. mangifera.

Proximate composition, fungal isolation and contamination of aflatoxin B1 in chickpea seeds from the Punjab, Pakistan.

Chickpea, Cicer arietinum L., is a nutrient rich crop that is widely cultivated and consumed in Pakistan. However, chickpea is highly prone to fungal growth leading to contamination with aflatoxins, the most potent carcinogen found in nature. In this study, fifty chickpea seed samples were collected from the local markets of the Punjab, Pakistan, to evaluate their nutritional quality, fungal and AFB1 contamination. Proximate analysis suggested that chickpea seeds contained 5.5-6.93% moisture, 62.24-63.24% carbohydrates, 22.75-23.44% protein, 4.99-5.4% fat, 5.62-5.84% fiber and 2.92-3.16% ash. Morphological identification techniques revealed fourteen fungal species belonging to six fungal genera from which Aspergillus flavus was the leading contaminant. AFB1 analysis revealed that sixty-two percent samples were contaminated with AFB1. All the AFB1 positive samples contained AFB1 level more than 2 ppb and 12.9% samples contain AFB1 level more than 20 ppb, exceeded the maximum limit (ML) assigned by EU and USA (FDA & FAO) respectively. The results of the present studies reported that chickpea is a highly contaminated commodity in terms of fungi and AFB1 that's why further investigations and monitoring are required to reduce the fungal and AFB1 contamination. These baseline data are an initial step in the effort to deal with this significant food safety issue.

Effect of Phytosynthesized Selenium and Cerium Oxide Nanoparticles on Wheat (Triticum aestivum L.) against Stripe Rust Disease.

In this study, selenium nanoparticles (SeNPs) and cerium oxide nanoparticles (CeONPs) were synthesized by using the extract of Melia azedarach leaves, and Acorus calamusas rhizomes, respectively, and investigated for the biological and sustainable control of yellow, or stripe rust, disease in wheat. The green synthesized NPs were characterized by UV-Visible spectroscopy, scanning electron microscopy (SEM), energy-dispersive X-ray (EDX), and X-ray diffraction (XRD). The SeNPs and CeONPs, with different concentrations (i.e., 10, 20, 30, and 40 mg/L), were exogenously applied to wheat infected with Puccinia striformis. SeNPs and CeONPs, at a concentration of 30 mg/L, were found to be the most suitable concentrations, which reduced the disease severity and enhanced the morphological (plant height, root length, shoot length, leaf length, and ear length), physiological (chlorophyll and membrane stability index), biochemical (proline, phenolics and flavonoids) and antioxidant (SOD and POD) parameters. The antioxidant activity of SeNPs and CeONPs was also measured. For this purpose, different concentrations (50, 100, 150, 200 and 400 ppm) of both SeNPs and CeONPs were used. The concentration of 400 ppm most promoted the DPPH, ABTS and reducing power activity of both SeNPs and CeONPs. This study is considered the first biocompatible approach to evaluate the potential of green synthesized SeNPs and CeONPs to improve the health of yellow, or stripe rust, infected wheat plants and to provide an effective management strategy to inhibit the growth of Puccinia striformis.

Comprehensive Review of Aflatoxin Contamination, Impact on Health and Food Security, and Management Strategies in Pakistan.

Aflatoxins (AFs) are the most important toxic, mutagenic, and carcinogenic fungal toxins that routinely contaminate food and feed. While more than 20 AFs have been identified to date, aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1), G2 (AFG2), and M1 (AFM1) are the most common. Over 25 species of Aspergillus have been shown to produce AFs, with Aspergillus flavus, Aspergillus parasiticus, and Aspergillus nomius being the most important and well-known AF-producing fungi. These ubiquitous molds can propagate on agricultural commodities to produce AFs in fields and during harvesting, processing, transportation, and storage. Countries with warmer climates and that produce foods susceptible to AF contamination shoulder a substantial portion of the global AF burden. Pakistan's warm climate promotes the growth of toxigenic fungi, resulting in frequent AF contamination of human foods and animal feeds. The potential for contamination in Pakistan is exacerbated by improper storage conditions and a lack of regulatory limits and enforcement mechanisms. High levels of AFs in common commodities produced in Pakistan are a major food safety problem, posing serious health risks to the population. Furthermore, aflatoxin contamination contributes to economic losses by limiting exports of these commodities. In this review, recent information regarding the fungal producers of AFs, prevalence of AF contamination of foods and feed, current regulations, and AF prevention and removal strategies are summarized, with a major focus on Pakistan.

Left ventricular assist device bioinformatics identify possible hubgenes and regulatory networks involved in the myocardium of patients with left ventricular assist device.

Objective: The aim of this study was to clarify the changes of myocardial gene expression profile after left ventricular assist device (LVAD) implantation and the related molecular biological significance. Methods: A thorough bioinformatic analysis to evaluate the changes in gene expression profile in patients pre-LVAD and post-LVAD was conducted. Four relevant gene expression datasets-GSE430, GSE974, GSE21610, and GSE52601 from Gene Expression Omnibus (GEO) database were downloaded. Analysis of GEO2R, Gene Ontology (GO), protein-protein interaction (PPI) were used to determine differentially expressed genes (DEGs) and their function, respectively. Results: A total of 37 DEGs were identified, including 26 down-regulated and 11 up-regulated genes. The molecular function of DEGs were enriched in "cytokine activity," "neurotransmitter binding," "receptor ligand activity." The gene set enrichment analysis (GSEA) revealed an overall marked increase of neutrophil degranulation signaling, closely correlated with the G protein coupled receptor (GPCR)-ligand binding process after LVAD assistance. 16 hubgenes in these DEGs were further selected and the biological process involved is mainly related to positive regulation of leukocyte chemotaxis mediated by chemokines. Conclusion: Inflammatory signaling pathway is crucial for the pathophysiology after LVAD implantation. Chemokines mediate cardiac inflammatory response and tissue remodeling after LVAD implantation through GPCR-ligand binding.

Impact of Patient Navigation on Population-Based Breast Screening: a Systematic Review and Meta-analysis of Randomized Clinical Trials.

BACKGROUND: Unsatisfactory cancer screening results are often associated with poor prognosis. This study synthesized the literatures addressing the impact of patient navigation (PN) interventions on population-based breast cancer screening promotion to identify characteristics of the model for addressing breast cancer disparities. METHODS: We searched Pubmed, Embase, Web of Science, and the Cochrane Central Registry from inception to 31 December 2020 for randomized controlled trials (PROSPERO: CRD42021246890). We double blindly abstracted data and assessed study quality. We assessed screening completion rates and diagnostic resolution using random-effects models between those receiving navigation and controls. RESULTS: Of 236 abstracts identified, 15 studies met inclusion criteria. Nine of the papers evaluated the impact of PN on breast screening, while the other six were on the resolution of abnormal screening results. Compared to the non-PN group, PN improved screening completion (OR: 2.0, 95% CI: 1.4-2.8]) and shortened the time to diagnosis (WMD: - 9.90 days, 95% CI: - 19.09 to - 0.71). CONCLUSIONS: Patient navigation improves breast cancer screening rates but does not improve resolution of abnormal tests.

Characterization of 260 Isolates of Aspergillus Section Flavi Obtained from Sesame Seeds in Punjab, Pakistan.

Sesame Sesamum indicum L. is a major oil-based seed crop that has been widely cultivated and consumed in Pakistan. Unfortunately, sesame is highly prone to Aspergillus fungal growth in the field, and under inappropriate storage conditions can become contaminated with aflatoxins, the most potent carcinogen found in nature. Here, we have isolated a high number of Aspergillus isolates from sesame seeds in fresh and stored conditions obtained from rainfed and irrigated zones of Punjab, Pakistan, and characterized them for aflatoxigenic potentials. Using morphological identification techniques, 260 isolates were grouped as potential Aspergillus section Flavi, with 126 and 134 originating from the rainfed and irrigated zones, respectively. Out of 260 in total, 188 isolates were confirmed to produce aflatoxins. There were no significant differences in potential aflatoxigenic isolates with respect to the rainfed and irrigated zones. However, the number of potential aflatoxigenic isolates was significantly higher (p < 0.05) in stored samples than that of those from fresh sesame seeds in the rainfed and irrigated zone. Whole genome sequencing and comparative analyses of 12 select isolates have revealed that one of the A. flavus isolates, which produced very low aflatoxins (AFP10), has an elevated missense variant rate, numerous high impact mutations, and a 600 base pair deletion in the norB gene. In summary, our study provides insights into aflatoxigenic potential and the associated genetic diversity of indigenous Aspergillus section Flavi isolates and potential management strategies for reducing aflatoxin contamination levels in a major crop consumed in Punjab, Pakistan.

Human Plasma Metabolomics Identify 9-cis-retinoic Acid and Dehydrophytosphingosine Levels as Novel biomarkers for Early Ventricular Fibrillation after ST-elevated Myocardial Infarction.

The relevant metabolite biomarkers for risk prediction of early onset of ventricular fibrillation (VF) after ST-segment elevation myocardial infarction (STEMI) remain unstudied. Here, we aimed to identify these imetabolites and the important metabolic pathways involved, and explore whether these metabolites could be used as predictors for the phenotype. Plasma samples were obtained retrospectively from a propensity-score matched cohort including 42 STEMI patients (21 consecutive VF and 21 non-VF). Ultra-performance liquid chromatography and mass spectrometry in combination with a comprehensive analysis of metabolomic data using Metaboanalyst 5.0 version were performed. As a result, the retinal metabolism pathway proved to be the most discriminative for the VF phenotype. Furthermore, 9-cis-Retinoic acid (9cRA) and dehydrophytosphingosine proved to be the most discriminative biomarkers. Biomarker analysis through receiver operating characteristic (ROC) curve showed the 2-metabolite biomarker panel yielding an area under the curve (AUC) of 0.836. The model based on Monte Carlo cross-validation found that 9cRA had the greatest probability of appearing in the predictive panel of biomarkers in the model. Validation of model efficiency based on an ROC curve showed that the combination model constructed by 9cRA and dehydrophytosphingosine had a good predictive value for early-onset VF after STEMI, and the AUC was 0.884 (95% CI 0.714-1). Conclusively, the retinol metabolism pathway was the most powerful pathway for differentiating the post-STEMI VF phenotype. 9cRA was the most important predictive biomarker of VF, and a plasma biomarker panel made up of two metabolites, may help to build a potent predictive model for VF.

The Potential Therapeutic Role of Celastrol in Patients With Heart Failure With Preserved Ejection Fraction.

Analysis of left ventricular systolic dysfunction remained at the centre of heart failure research for many years (also known as heart failure with reduced ejection fraction, HFrEF). Although more than 50% of all heart failure patients experience a form of heart failure characterised by preserved ejection fraction (HFpEF), the pathophysiological mechanisms leading to this form of heart failure remain not well-understood. Several evidence-based treatments for HFrEF are in routine use, but there are limited evidence-based therapies for HFpEF. The effects of these remain controversial, with current treatment options being limited to managing the associated symptoms and conditions. Accumulating evidence demonstrates that pro-inflammatory and oxidative stress pathways play key roles in the development and progression of HFpEF, such as the Unfolded Protein Response (UPR) and inducible nitric oxide synthase. Celastrol, derived from medicinal plants, is a bioactive compound with strong anti-inflammatory properties, which could deem it as fruitful in overcoming the effects of such dysregulated UPR. This literature review therefore focuses on Celastrol's anti-inflammatory and antioxidant activities, alongside its other potential therapeutic activities, and its ability to impede the pathways that are thought to be involved in the development of HFpEF, such as the JAK2/STAT pathway, to elucidate the potential therapeutic role of this bioactive compound, in the treatment of HFpEF.

Mycobiota Isolation and Aflatoxin B1 Contamination in Fresh and Stored Sesame Seeds from Rainfed and Irrigated Zones of Punjab, Pakistan.

ABSTRACT: This study was carried out to evaluate the prevalence of mycobiota and aflatoxin (AF) B1 contamination in sesame seeds from rainfed and irrigated zones of the Punjab, Pakistan. For this purpose, 100 sesame seed samples were collected directly from the fields of major sesame-producing areas in rainfed and irrigated zones. The agar plate method was used for isolation of mycobiota, and thin-layer chromatography was used to determine AFB1 concentrations. Seed samples were then stored for 12 months. After 12 months, the seeds were again analyzed for mycobiota and AFB1 for comparison. All samples were positive for fungal growth under fresh and stored conditions. Twenty-one fungal species of 10 genera were isolated. Aspergillus flavus was the most prevalent contaminant found in fresh and stored sesame seeds from rainfed and irrigated zones, followed by Aspergillus niger, Alternaria alternata, and Fusarium oxysporum. The least prevalent fungi were Aspergillus ochraceus and Cladosporium oxysporum. Analysis revealed that 92% of fresh and 99% of stored seed samples were contaminated with AFB1. In the rainfed zone, 88% of fresh and 100% of stored seed samples were contaminated with AFB1, with mean concentrations of 15.74 and 33.8 ppb, respectively. In the irrigated zone, 96% of fresh and 98% of stored seed samples were contaminated with AFB1, with mean concentrations of 20.5 and 27.56 ppb, respectively. AFB1 concentrations >20 ppb were found in 20% of fresh and 100% of stored seeds samples from the rainfed zone and in 28% of fresh and 60% of stored samples from the irrigated zone and thus were not fit for human consumption as per the maximum limit (20 ppb) assigned by the U.S. Food and Drug Administration and the Food and Agriculture Organization of the United Nations. This report is the first on the mycobiota and AFB1 contamination in sesame seeds from rainfed and irrigated zones of the Punjab, Pakistan. These baseline data are an initial step in the effort to deal with this significant food safety issue.

Designing indaceno thiophene-based three new molecules containing non-fullerene acceptors as strong electron withdrawing groups with DFT approaches.

Nowadays, scientists are trying to develop low-cost fullerene free acceptors for small organic photovoltaic cells in order to overcome the limitations of fullerene derivatives. Current research work deals with theoretical study on three non-fullerene acceptors based on indaceno, dithiophene core, and thiophene bridge units linked with dissimilar end non-fullerene groups which act as strong acceptor moieties. Different optoelectronic characteristics of the designed molecules were calculated and compared with the reference compound R (indaceno dithiophene-based fused ring acceptor) which is recently reported. Results shows that C2 and C3 exhibit broad absorption spectrum and lower band gap whereas C2 and C1 exhibit highest open-circuit voltages VOC value with B3LYP and MPW1PW91 functionals respectively as compared with the R. All designed molecules have high dipole moment values, lower value of hole reorganization energy λh than electron reorganization energy λe which reflects that our designed acceptor molecules are good candidates for organic photovoltaics. Graphical abstract Absorption spectra of R and three designed non-fullerene acceptors with strong absorption band in the visible region of solar cells spectrum.