The proteomic landscape of genotoxic stress-induced micronuclei.

Micronuclei (MN) are induced by various genotoxic stressors and amass nuclear- and cytoplasmic-resident proteins, priming the cell for MN-driven signaling cascades. Here, we measured the proteome of micronuclear, cytoplasmic, and nuclear fractions from human cells exposed to a panel of six genotoxins, comprehensively profiling their MN protein landscape. We find that MN assemble a proteome distinct from both surrounding cytoplasm and parental nuclei, depleted of spliceosome and DNA damage repair components while enriched for a subset of the replisome. We show that the depletion of splicing machinery within transcriptionally active MN contributes to intra-MN DNA damage, a known precursor to chromothripsis. The presence of transcription machinery in MN is stress-dependent, causing a contextual induction of MN DNA damage through spliceosome deficiency. This dataset represents a unique resource detailing the global proteome of MN, guiding mechanistic studies of MN generation and MN-associated outcomes of genotoxic stress.

A clinically applicable integrative molecular classification of meningiomas.

Meningiomas are the most common primary intracranial tumour in adults1. Patients with symptoms are generally treated with surgery as there are no effective medical therapies. The World Health Organization histopathological grade of the tumour and the extent of resection at surgery (Simpson grade) are associated with the recurrence of disease; however, they do not accurately reflect the clinical behaviour of all meningiomas2. Molecular classifications of meningioma that reliably reflect tumour behaviour and inform on therapies are required. Here we introduce four consensus molecular groups of meningioma by combining DNA somatic copy-number aberrations, DNA somatic point mutations, DNA methylation and messenger RNA abundance in a unified analysis. These molecular groups more accurately predicted clinical outcomes compared with existing classification schemes. Each molecular group showed distinctive and prototypical biology (immunogenic, benign NF2 wild-type, hypermetabolic and proliferative) that informed therapeutic options. Proteogenomic characterization reinforced the robustness of the newly defined molecular groups and uncovered highly abundant and group-specific protein targets that we validated using immunohistochemistry. Single-cell RNA sequencing revealed inter-individual variations in meningioma as well as variations in intrinsic expression programs in neoplastic cells that mirrored the biology of the molecular groups identified.

Targeted Mass Spectrometry of Longitudinal Patient Sera Reveals LTBP1 as a Potential Surveillance Biomarker for High-Grade Serous Ovarian Carcinoma.

High-grade serous ovarian carcinoma (HGSC) is the most prevalent subtype of epithelial ovarian cancer. The combination of a high rate of recurrence and novel therapies in HGSC necessitates an accurate assessment of the disease. Currently, HGSC response to treatment and recurrence are monitored via immunoassay of serum levels of the glycoprotein CA125. CA125 levels predictably rise at HGSC recurrence; however, it is likely that the disease is progressing even before it is detectable through CA125. This may explain why treating solely based on CA125 increase has not been associated with improved outcomes. Thus, additional biomarkers that monitor HGSC progression and cancer recurrence are needed. For this purpose, we developed a scheduled parallel reaction monitoring mass spectrometry (PRM-MS) assay for the quantification of four previously identified HGSC-derived glycopeptides (from proteins FGL2, LGALS3BP, LTBP1, and TIMP1). We applied the assay to quantify their longitudinal expression profiles in 212 serum samples taken from 34 HGSC patients during disease progression. Analyses revealed that LTBP1 best-mirrored tumor load, dropping as a result of cancer treatment in 31 out of 34 patients and rising at HGSC recurrence in 28 patients. Additionally, LTBP1 rose earlier during remission than CA125 in 11 out of 25 platinum-sensitive patients with an average lead time of 116.4 days, making LTBP1 a promising candidate for monitoring of HGSC recurrence.

Comprehensive Prostate Fluid-Based Spectral Libraries for Enhanced Protein Detection in Urine.

Biofluids contain molecules in circulation and from nearby organs that can be indicative of disease states. Characterizing the proteome of biofluids with DIA-MS is an emerging area of interest for biomarker discovery; yet, there is limited consensus on DIA-MS data analysis approaches for analyzing large numbers of biofluids. To evaluate various DIA-MS workflows, we collected urine from a clinically heterogeneous cohort of prostate cancer patients and acquired data in DDA and DIA scan modes. We then searched the DIA data against urine spectral libraries generated using common library generation approaches or a library-free method. We show that DIA-MS doubles the sample throughput compared to standard DDA-MS with minimal losses to peptide detection. We further demonstrate that using a sample-specific spectral library generated from individual urines maximizes peptide detection compared to a library-free approach, a pan-human library, or libraries generated from pooled, fractionated urines. Adding urine subproteomes, such as the urinary extracellular vesicular proteome, to the urine spectral library further improves the detection of prostate proteins in unfractionated urine. Altogether, we present an optimized DIA-MS workflow and provide several high-quality, comprehensive prostate cancer urine spectral libraries that can streamline future biomarker discovery studies of prostate cancer using DIA-MS.

Slalom technique for multilevel degenerative spinal stenosis, surgical outcome and complication rate: Review of our experience.

Lumbar canal stenosis (LCS) is a common spinal disease affecting the elderly. Primarily it is asymptomatic until there is neurogenic claudication. Minimally invasive surgical (MIS) techniques are used to treat patients with lumbar spinal stenosis (LSS), while tubular system with alternative multilevel decompression is specifically used for those with minimal back pain and no mechanical instability on dynamic imaging. The aim of the study is to evaluate surgical outcome of Slalom procedure and complications in Middle East population. One hundred and five patients with lumbar stenosis (61 males and 44 females) underwent the procedure between 2015-2021 who were regularly followed-up using preoperative and postoperative COMI score (the core outcome measure index) at six months after index surgery. Progressive improvement in COMI score from average seven pre-op score to an average of three after six months of index surgery. The postoperative complications were dural tear (6.67%), Postoperative infection (3.81%), mechanical instability (1.9%), postoperative neuritis (8.57%) and death (1.9%).

Phosphoproteomic Analysis of Dopamine D2 Receptor Signaling Reveals Interplay of G Protein- and ß-Arrestin-Mediated Effects.

Leveraging biased signaling of G protein-coupled receptors has been proposed as a promising strategy for the development of drugs with higher specificity. However, the consequences of selectively targeting G protein- or ß-arrestin-mediated signaling on cellular functions are not comprehensively understood. In this study, we utilized phosphoproteomics to gain a systematic overview of signaling induced by the four biased and balanced dopamine D2 receptor (D2R) ligands MS308, BM138, quinpirole, and sulpiride in an in vitro D2R transfection model. Quantification of 14,160 phosphosites revealed a low impact of the partial G protein agonist MS308 on cellular protein phosphorylation, as well as surprising similarities between the balanced agonist quinpirole and the inverse agonist sulpiride. Analysis of the temporal profiles of ligand-induced phosphorylation events showed a transient impact of the G protein-selective agonist MS308, whereas the ß-arrestin-preferring agonist BM138 elicited a delayed, but more pronounced response. Functional enrichment analysis of ligand-impacted phosphoproteins and treatment-linked kinases confirmed multiple known functions of D2R signaling while also revealing novel effects, for example of MS308 on sterol regulatory element-binding protein-related gene expression. All raw data were deposited in MassIVE (MSV000089457).

Combination of inhibitors for two glycolytic enzymes portrays high synergistic efficacy against Cryptosporidium parvum.

Cryptosporidium is an intracellular protozoan parasite that causes serious enteric disease in humans and in a wide range of animals worldwide. Despite its high prevalence, no effective therapeutic drugs are available against life-threatening cryptosporidiosis in at-risk populations including malnourished children, immunocompromised patients, and neonatal calves. Thus, new efficacious drugs are urgently needed to treat all susceptible populations with cryptosporidiosis. Unlike other apicomplexans, Cryptosporidium parvum lacks the tricarboxylic acid cycle and the oxidative phosphorylation steps, making it solely dependent on glycolysis for metabolic energy production. We have previously reported that individual inhibitors of two unique glycolytic enzymes, the plant-like pyruvate kinase (CpPyK) and the bacterial-type lactate dehydrogenase (CpLDH), are effective against C. parvum, both in vitro and in vivo. Herein, we have derived combinations of CpPyK and CpLDH inhibitors with strong synergistic effects against the growth and survival of C. parvum, both in vitro and in an infection mouse model. In infected immunocompromised mice, compound combinations of NSC303244 + NSC158011 and NSC252172 + NSC158011 depicted enhanced efficacy against C. parvum reproduction and ameliorated intestinal lesions of cryptosporidiosis at doses fourfold lower than the total effective doses of individual compounds. Importantly, unlike individual compounds, NSC303244 + NSC158011 combination was effective in clearing the infection completely without relapse in immunocompromised mice. Collectively, our study has unveiled compound combinations that simultaneously block two essential catalytic steps for metabolic energy production in C. parvum to achieve improved efficacy against the parasite. These combinations are, therefore, lead compounds for the development of a new generation of efficacious anti-cryptosporidial drugs.

Correlation of brain-derived neurotrophic factor with severity of depression and treatment response.

Background: Brain-derived neurotrophic factor (BDNF) is a modulator of neuroplasticity in the brain. It plays an important role in the pathophysiology of depression through the stress pathway. The information about correlation of BDNF levels with depression severity and treatment response in Indian population is scarce. Methods: Consecutive 60 never treated cases with depression reporting to a large tertiary care psychiatry unit and 60 healthy matched controls from 01 January 2016 to 31 December 2016 were enrolled for study. Sociodemographic data were collected. Diagnosis of depression was carried out as per International Classification of Diseases-10th revision (ICD-10) diagnostic criteria for research. The Hamilton Rating Scale for Depression (HRSD) was administered and accordingly scored. Venous blood for BDNF levels was collected from all cases and controls. Cases were reassessed after 04 weeks of treatment with HRSD and BDNF levels. Results: The mean level of serum BDNF among cases (18.56 ng/ml) was found to be reduced significantly as compared with healthy controls (32.41 ng/ml). The mean serum BDNF level (18.56 ng/ml) in never treated cases was significantly negatively correlated with the median clinical HRSD score (18.5). There was a significant increase in the mean level of serum BDNF after antidepressant treatment. Conclusion: The study has revealed statistically significant low levels of serum BDNF in cases not exposed to treatment with depression compared with healthy controls. There was significant negative correlation of levels of serum BDNF with depression severity. The levels of serum BDNF significantly increased after four weeks of treatment.

The proteomic landscape of glioblastoma recurrence reveals novel and targetable immunoregulatory drivers.

Glioblastoma (GBM) is characterized by extensive cellular and genetic heterogeneity. Its initial presentation as primary disease (pGBM) has been subject to exhaustive molecular and cellular profiling. By contrast, our understanding of how GBM evolves to evade the selective pressure of therapy is starkly limited. The proteomic landscape of recurrent GBM (rGBM), which is refractory to most treatments used for pGBM, are poorly known. We, therefore, quantified the transcriptome and proteome of 134 patient-derived pGBM and rGBM samples, including 40 matched pGBM-rGBM pairs. GBM subtypes transition from pGBM to rGBM towards a preferentially mesenchymal state at recurrence, consistent with the increasingly invasive nature of rGBM. We identified immune regulatory/suppressive genes as important drivers of rGBM and in particular 2-5-oligoadenylate synthase 2 (OAS2) as an essential gene in recurrent disease. Our data identify a new class of therapeutic targets that emerge from the adaptive response of pGBM to therapy, emerging specifically in recurrent disease and may provide new therapeutic opportunities absent at pGBM diagnosis.

Anti-cancer medicine shortages in an oncology tertiary hospital of Pakistan: A five-year retrospective study.

OBJECTIVE: Anti-cancer medicine shortages are advancing challenges for patients and hospitals. This study aims to evaluate anti-cancer and supportive medicine shortages in a tertiary hospital in Pakistan and propose solutions. METHOD: A retrospective observational research was performed in a tertiary care hospital in Pakistan from 2016 to 2020. Data was retrieved from the hospital database using a questionnaire regarding short medicines' generic name, brand, dosage, source, total source, frequency, causes, impact, management, and analyzed by Microsoft Excel 2013. RESULTS: Between January 2016 and December 2020, 43 individual medicine shortages were observed, with an average of 8.6 shortages per year. There were shortages of 22 medicines, including 8 anti-cancer (36.4%) and 14 supportive agents (63.6%). Total shortage days were 27,100, with an average of 1232 days (SD 757) per medicine. Supportive medicines' shortages were frequent, but oncology agents' shortages were constant. The most affected dosage form was injection. Cardiovascular drugs and alkylating agents were the most affected class in supportive and anti-cancer medicines, respectively. The use of "alternative medicine" and "patient needs based importation" were the most common mitigation strategies. CONCLUSION: Shortages of oncology medicines are challenging in Pakistan. The most prominent causes are the lack of updated governmental regulations, registration, and import issues. The tertiary care hospital has very few sources of supply, so it imports these drugs on a need basis to manage the shortages. But it is still concerning because of the huge financial burden on patients and institutions due to expensive import, and therapy become delayed as the import process takes time. Moreover, the most affected drug class was alkylating agents, and dosage was both injectable and oral medicines.

Regression of Hepatic Fibrosis and Evolution of Cirrhosis: A Concise Review.

Fibrosis is not a unidirectional, linear process, but a dynamic one resulting from an interplay of fibrogenesis and fibrolysis depending on the extent and severity of a biologic insult, or lack thereof. Regression of fibrosis has been documented best in patients treated with phlebotomies for hemochromatosis, and after successful suppression and eradication of chronic hepatitis B and C infections. This evidence mandates a reconsideration of the term "cirrhosis," which implies an inevitable progression towards liver failure. Furthermore, it also necessitates a staging system that acknowledges the bidirectional nature of evolution of fibrosis, and has the ability to predict if the disease process is progressing or regressing. The Beijing classification attempts to fill this gap in contemporary practice. It is based on microscopic features termed "the hepatic repair complex," defined originally by Wanless and colleagues. The elements of the hepatic repair complex represent the 3 processes of fragmentation and regression of scar, vascular remodeling (resolution), and parenchymal regeneration. However, regression of fibrosis does not imply resolution of cirrhosis, which is more than just a stage of fibrosis. So far, there is little to no evidence to suggest that large regions of parenchymal extinction can be repopulated by regenerating hepatocytes. Similarly, the vascular lesions of cirrhosis persist, and there is no evidence of complete return to normal microcirculation in cirrhotic livers. In addition, the risk of hepatocellular carcinoma is higher compared with the general population and these patients need continued screening and surveillance.

Efficacy of chitosan-coated textile waste biochar applied to Cd-polluted soil for reducing Cd mobility in soil and its distribution in moringa (Moringa oleifera L.).

Soil pollution with Cd has promoted serious concerns for medicinal plant quality. Amending Cd-polluted soils with textile waste biochar (TWB) coated with natural polymers can lower Cd bioavailability in them and reduce associated environmental and human health risks. In this study, we explored the impacts of solely applied TWB, chitosan (CH), their mix (TWB + CH) and TWB coated with CH (TBC) in Cd-polluted soil on Cd distribution in moringa (Moringa oleifera L.) shoots and roots as well as plant-available Cd in soil. Moreover, amendments effects on plant growth, dietary quality, and antioxidative defense responses were also assessed. Results revealed that the addition of TWB, CH, and TWB + CH in Cd-polluted soil reduced Cd distribution in shoots (56%, 66%, and 63%), roots (41%, 48%, and 45%), and plant-available Cd in soil (38%, 52%, and 49%), compared to control. Interestingly, the TBC showed significantly the topmost response for reducing Cd concentrations in shoots, roots, and soil by 73%, 54%, and 58%, respectively, relative to control. Moreover, amending Cd-polluted soil with TWB, CH, and TWB + CH depicted significantly better effects on plant growth, dietary quality, and activities of soil enzymes but the topmost response was observed with TBC treatment. Compared with control, TBC improved plant growth parameters: shoot length (81%), root length (90%), shoot fresh weight (60%), root fresh weight (76%), shoot dry weight (75%), root dry weight (68%) contents of chlorophyll-a (42%) and chlorophyll-b (74%), and soil enzyme activities: urease (130%), catalase (138%), protease (71%), cellobiohydrolase (45%), acid phosphatase (34%), peroxidase (60%), ß-glucosidase (152%), chitinase (62%), and phosphomonoesterase (139%). Furthermore, TBC treatment arrested Cd-induced oxidative stress via escalating the activities of antioxidant enzymes as well as improved moringa dietary parameters (protein, tannins, lipids, alkaloids, saponins, terpenoids, flavonoids, and tocopherols contents). Such findings suggest that the TBC has an immense perspective to remediate Cd-polluted soils and prevent human health risks associated with Cd exposure through the diet.

Soil fertility, chemical properties, and pollutant removal efficiency of Salicornia europaea in response to different times and duration of wastewater irrigation.

Halophytes are the good candidates in coastal saline areas which could be irrigated with wastewater. The purpose of this study was to evaluate the soil-water-plant system under control and wastewater irrigation (containing toxic elements and organic matter) at three durations (vegetative, flowering, and reproductive stages) and two exposure times (2 and 4 days in each stage). The results obtained in the experimental tests for wastewater irrigation indicated that the Salicornia is efficient for the removal of chemical oxygen demand (61%), biochemical oxygen demand (74%), total suspended solids (47.6%), and ammoniacal nitrogen (64%) at the reproductive stage. At the same time, the average nitrate concentration increased to 51.3 mg L-1 with more solids. Regardless of wastewater irrigation duration, irrigation with wastewater significantly increased organic matter, nitrogen, phosphorus, and potassium of the soil. The Mg2+ and Ca2+ contents in the aboveground biomass of the plants were also high ranged from 0.58 to 1%, and 0.43 to 0.68 mg g-1 DW, respectively. All the exchangeable cations other than Na+ were higher for wastewater irrigation at the flowering stage. Plants maintained noticeably higher Ca2+/Na+ and K+/Na+ ratios in the roots than those in the shoots except for 4 days after the reproductive stage. S. europaea is well adapted to grow in wastewater irrigation and can tolerate hypoxic conditions through improving water and soil quality.

Recent advances in mass spectrometry based clinical proteomics: applications to cancer research.

Cancer biomarkers have transformed current practices in the oncology clinic. Continued discovery and validation are crucial for improving early diagnosis, risk stratification, and monitoring patient response to treatment. Profiling of the tumour genome and transcriptome are now established tools for the discovery of novel biomarkers, but alterations in proteome expression are more likely to reflect changes in tumour pathophysiology. In the past, clinical diagnostics have strongly relied on antibody-based detection strategies, but these methods carry certain limitations. Mass spectrometry (MS) is a powerful method that enables increasingly comprehensive insights into changes of the proteome to advance personalized medicine. In this review, recent improvements in MS-based clinical proteomics are highlighted with a focus on oncology. We will provide a detailed overview of clinically relevant samples types, as well as, consideration for sample preparation methods, protein quantitation strategies, MS configurations, and data analysis pipelines currently available to researchers. Critical consideration of each step is necessary to address the pressing clinical questions that advance cancer patient diagnosis and prognosis. While the majority of studies focus on the discovery of clinically-relevant biomarkers, there is a growing demand for rigorous biomarker validation. These studies focus on high-throughput targeted MS assays and multi-centre studies with standardized protocols. Additionally, improvements in MS sensitivity are opening the door to new classes of tumour-specific proteoforms including post-translational modifications and variants originating from genomic aberrations. Overlaying proteomic data to complement genomic and transcriptomic datasets forges the growing field of proteogenomics, which shows great potential to improve our understanding of cancer biology. Overall, these advancements not only solidify MS-based clinical proteomics' integral position in cancer research, but also accelerate the shift towards becoming a regular component of routine analysis and clinical practice.

Role of sepiolite for cadmium (Cd) polluted soil restoration and spinach growth in wastewater irrigated agricultural soil.

Metals that contaminate soil are one of the major problems seriously affecting sustainable agriculture worldwide. Cadmium (Cd) toxicity to agricultural crops is a global problem. Mobility of Cd in contaminated soil can be minimized by the amendment of soil passivators which will ultimately reduce its movement from soil to plants. A pot study was performed to evaluate the impact of sepiolite from 1% to 5% on Cd solubility and its accumulation in spinach tissues. Soil pH, Cd fractionation, Cd accumulation in spinach tissue and Cd adsorption mechanism were determined. Results were recorded that soil pH was increased from 0.3 to 1.0 units with the increasing rate of sepiolite from 1% to 5%. Similarly, Cd contents in acid soluble phase was decreased by 42.8% and increased in residual phase by 35.8% at 5% rate, relative to control. Moreover, the significant reduction in Cd uptake by spinach shoots and roots was occurred by 26.2% and 30.6% at 5% rate, respectively. Furthermore, the maximum Cd adsorption capacity 37.35 mg g-1 was recorded at 5% rate relative to control. The analysis of FTIR, XRD and SEM also confirm the ability of sepiolite for Cd polluted soil restoration and thereby, reduces its phytoavailability in polluted soil to alleviate food security challenges.

Zinc finger protein transcription factors: Integrated line of action for plant antimicrobial activity.

The plants resist/tolerate unfavorable conditions in their natural habitats by using different but aligned and integrated defense mechanisms. Such defense responses include not only morphological and physiological adaptations but also the genomic and transcriptomic reconfiguration. Microbial attack on plants activates multiple pro-survival pathways such as transcriptional reprogramming, hypersensitive response (HR), antioxidant defense system and metabolic remodeling. Up-regulation of these processes during biotic stress conditions directly relates with plant survival. Over the years, hundreds of plant transcription factors (TFs) belonging to diverse families have been identified. Zinc finger protein (ZFP) TFs have crucial role in phytohormone response, plant growth and development, stress tolerance, transcriptional regulation, RNA binding and protein-protein interactions. Recent research progress has revealed regulatory and biological functions of ZFPs in incrementing plant resistance to pathogens. Integration of transcriptional activity with metabolic modulations has miniaturized plant innate immunity. However, the precise roles of different zinc finger TFs in plant immunity to pathogens have not been thoroughly analyzed. This review consolidates the pivotal functioning of zinc finger TFs and proposes the integrative understanding as foundation for the plant growth and development including the stress responses.

Development of Drought-Tolerant Transgenic Wheat: Achievements and Limitations.

Crop yield improvement is necessary to keep pace with increasing demand for food. Due to climatic variability, the incidence of drought stress at crop growth stages is becoming a major hindering factor to yield improvement. New techniques are required to increase drought tolerance along with improved yield. Genetic modification for increasing drought tolerance is highly desirable, and genetic engineering for drought tolerance requires the expression of certain stress-related genes. Genes have been identified which confer drought tolerance and improve plant growth and survival in transgenic wheat. However, less research has been conducted for the development of transgenic wheat as compared to rice, maize, and other staple food. Furthermore, enhanced tolerance to drought without any yield penalty is a major task of genetic engineering. In this review, we have focused on the progress in the development of transgenic wheat cultivars for improving drought tolerance and discussed the physiological mechanisms and testing of their tolerance in response to inserted genes under control or field conditions.

Effective Role of Biochar, Zeolite and Steel Slag on Leaching Behavior of Cd and Its Fractionations in Soil Column Study.

Remediation of cadmium (Cd) from contaminated soils is considered a complicated task of environmental safety. A column leaching experiment was planned to estimate the influence of biochar (BC), zeolite (ZE) and steel slag (SL) at 1.5% and 3% application rate on Cd leaching behavior and chemical fractionation in contaminated soil. A sequential extraction procedure, the European Community Bureau of Reference (BCR), Toxicity Characteristic Leaching Procedure (TCLP) and NH4NO3 were performed after leaching was completed. The soluble portion of Cd was decreased by 36.3%, 18.4% and 28.7% and Cd contents in leachate were decreased by 44.8%, 30% and 31.3% after BC, ZE and SL addition at 3% rate, respectively over control soil. The greater reduction in TCLP extractable Cd was observed by 29.6% with BC and 22.4% with ZE and 25.7% with SL at 3% application rate. Overall, biochar can be considered an efficient soil amendment to reduce Cd leaching as well as increased its stabilization within soil profile.

The response of transgenic Brassica species to salt stress: a review.

Salt stress is considered one of the main abiotic factors to limit crop growth and productivity by affecting morpho-physiological and biochemical processes. Genetically, a number of salt tolerant Brassica varieties have been developed and introduced, but breeding of such varieties is time consuming. Therefore, current focus is on transgenic technology, which plays an important role in the development of salt tolerant varieties. Various salt tolerant genes have been characterized and incorporated into Brassica. Therefore, such genetic transformation of Brassica species is a significant step for improvement of crops, as well as conferring salt stress resistance qualities to Brassica species. Complete genome sequencing has made the task of genetically transforming Brassica species easier, by identifying desired candidate genes. The present review discusses relevant information about the principles which should be employed to develop transgenic Brassica species, and also will recommend tools for improved tolerance to salinity.

Promoting the productivity and quality of brinjal aligned with heavy metals immobilization in a wastewater irrigated heavy metal polluted soil with biochar and chitosan.

Depleting aquifers, lack of planning and low socioeconomic status of Pakistani farmers have led them to use wastewater (WW) for irrigating their crops causing food contamination with heavy metals and ultimately negative effects on human health. This study evaluates the effects of chitosan (CH) and biochar (BC) on growth and nutritional quality of brinjal plant together with in situ immobilization of heavy metals in a soil polluted with heavy metals due to irrigation with wastewater (SPHIW) and further irrigated with the same WW. Both CH and BC were applied at three different rates i.e. low rate [(LR), BC0.5%, CH0.5% and BC0.25%+CH0.25%], medium rate [(MR), BC1%, CH1% and BC0.5%+CH0.5%] and high rate [(HR), BC1.5%, CH1.5% and BC0.75%+CH0.75%]. Result revealed that brinjal growth, antioxidant enzymes, and fruit nutritional quality significantly improved from LR to HR for each amendment, relative to control. However, these results were more prominent with BC alone and BC+CH, compared with CH alone at each rate. Similarly, with few exceptions, significant reduction in Ni, Cd, Co, Cr and Pb concentrations in the root, shoot and fruit were found in sole CH treatment both at LR and MR but in both CH and BC+CH treatments at HR, relative to control. Interestingly, the concentrations of Fe in the roots, shoots and fruit were more pronounced at BC treatments relative to CH and BC+CH treatments at each rate, compared to control. Overall, the BC+CH treatment at HR was the most effective treatment for in situ immobilization of heavy metals in SPHIW and further irrigated with the same WW, compared to rest of the treatments. This study indicates that BC0.75%+CH0.75% treatment can be used to reduce mobility and bioavailability of heavy metals in SPHIW and facilitates plant growth by improving the antioxidant system. However, the feasibility of BC0.75%+CH0.75% treatment should also be tested at the field scale.