Publications by authors named "Pooja Sharma"

191 Publications

Next-Generation Sequencing for Congenital Nephrotic Syndrome: A Multi-Center Cross-Sectional Study from India.

Indian Pediatr 2021 May;58(5):445-451

Division of Nephrology, Department of Pediatrics, All India Institute of Medical Sciences, New Delhi; India. Correspondence to: Professor Arvind Bagga, Division of Nephrology, Department of Pediatrics, All India Institute of Medical Sciences, New Delhi, India.

Objective: Information on etiology of congenital nephrotic syndrome in non-Caucasian populations is limited. This study aimed to determine the genetic basis of congenital nephrotic syndrome in Indian patients.

Methods: In this observational, cross-sectional study, whole exome sequencing was performed on samples from all children diagnosed with congenital nephrotic syndrome, presenting at centers collaborating in a nationwide registry and biorepository. Analysis was targeted to focus on reported or novel, pathogenic or likely pathogenic variants in 89 genes implicated in etiology of nephrotic syndrome. Sanger sequencing was used to confirm disease-causing variants in patients and allelic segregation of compound heterozygous variants in samples from parents. Inheritance of a shared haplotype was analyzed among ten individuals carrying the most common variant.

Results: During 2017-2019, 34 patients with congenital nephrotic syndrome were screened. Consanguinity and similar illness in siblings were reported in eleven patients each. Homozygous or compound heterozygous, pathogenic or likely pathogenic variants were found in NPHS1 in 24 cases, including two novel variants. One patient each had homozygous pathogenic or likely pathogenic known or novel variant in NPHS2, PLCE1, OSGEP and LAMB2 genes. Patients with OSGEP and LAMB2 mutations had phenotype typical of Galloway Mowat and Pierson syndromes, respectively. Three variants in NPHS1 were common to 16 individuals. One reported variant in exon 19 (c.2600G>A; p.Gly867Asp) appears to share a common founder.

Conclusions: A genetic cause was determined for 82.4% patients with congenital nephrotic syndrome. Variants in NPHS1 are most common in Indian patients and founder mutations might be present.
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May 2021

An overview of latest advances in exploring bioactive peptide hydrogels for neural tissue engineering.

Biomater Sci 2021 May 11. Epub 2021 May 11.

Institute of Nano Science and Technology, Sector 81, Knowledge city, Mohali, 140306, Punjab, India.

Neural tissue engineering holds great potential in addressing current challenges faced by medical therapies employed for the functional recovery of the brain. In this context, self-assembling peptides have gained considerable interest owing to their diverse physicochemical properties, which enable them to closely mimic the biophysical characteristics of the native ECM. Additionally, in contrast to synthetic polymers, which lack inherent biological signaling, peptide-based nanomaterials could be easily designed to present essential biological cues to the cells to promote cellular adhesion. Moreover, injectability of these biomaterials further widens their scope in biomedicine. In this context, hydrogels obtained from short bioactive peptide sequences are of particular interest owing to their facile synthesis and highly tunable properties. In spite of their well-known advantages, the exploration of short peptides for neural tissue engineering is still in its infancy and thus detailed discussion is required to evoke interest in this direction. This review provides a general overview of various bioactive hydrogels derived from short peptide sequences explored for neural tissue engineering. The review also discusses the current challenges in translating the benefits of these hydrogels to clinical practices and presents future perspectives regarding the utilization of these hydrogels for advanced biomedical applications.
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http://dx.doi.org/10.1039/d0bm02049dDOI Listing
May 2021

Distillery wastewater detoxification and management through phytoremediation employing Ricinus communis L.

Bioresour Technol 2021 Apr 20;333:125192. Epub 2021 Apr 20.

Department of Environmental Microbiology, Babasaheb Bhimrao Ambedkar Central University, Vidya Vihar, Raebareli Road, Lucknow 226025, UP, India. Electronic address:

This study aimed to assess the phytoremediation potential of Ricinus communis L. for heavy metals remediation via rhizospheric bacterial activities for distillery wastewater detoxification and management. Results revealed that distillery wastewater contained high levels of metals and other physico-chemical pollution parameters that could cause environmental pollution and aquatic toxicity. The identified bacterium produced several plant growth-promoting compounds including siderophores, ligninolytic enzymes, and indole acetic acid that resulted in nutrient enhancement and improved mineralization of metals in the plants during stress conditions. The bioconcentration factor (BCF) of all the metals examined were > 1, which showed that these metals are accumulating in the root, shoot, and leaves of Ricinus communis L. Most of the metals are stablised in the roots but Pb, Cd and Zn were translocated more to the shoorts (TC>1). The ability of Ricinus communis L. to grow in metals-containing distillery wastewater and reduce heavy metals and organic contaminants suggests that it can be used to provide an effective treatment of distillery wastewater. The use of Ricinus communis L. is an eco-friendly tool for the reduction of organometallic contamination and protecting agricultural land.
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http://dx.doi.org/10.1016/j.biortech.2021.125192DOI Listing
April 2021

Naringenin mitigates behavioral alterations and provides neuroprotection against 3-nitropropinoic acid-induced Huntington's disease like symptoms in rats.

Nutr Neurosci 2021 Apr 15:1-11. Epub 2021 Apr 15.

Department of Toxicology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, India.

Background: Naringenin is a powerful antioxidant and anti-inflammatory flavonoid which has been widely used as a therapeutic agent in various toxic models. However, few studies have clearly discussed the neuromodulatory effects of naringenin against different neurodegenerative disorders.

Aim: We investigated the neuroprotective efficacy of naringenin against 3-nitropropionic acid (3-NP)-induced neurobehavioral, biochemical and histopathological alterations in rats.

Methods: Albino Wistar rats were randomly divided into three experimental groups. Group 1, the vehicle administered group, received saline. Group 2 received 3-NP (20 mg/kg body weight, ) for 4 consecutive days. Group 3 received naringenin (50 mg/kg body weight, p.o.) twice daily for a period of 4 days, 30 min before and 6 h after the 3-NP administration. On the 5th day, neurobehavioral experiments were performed to access the behavioral outcomes and the striatum tissue was used for analysis of the monoamine oxidase (MAO) activity and serotonin (5-HT) levels. In addition, astrocytes activation was observed by glial fibrillary acidic protein (GFAP) immunostaining.

Results: Our results showed that naringenin co-treatment provides neuroprotection against 3-NP-induced neurological disorders. Naringenin also increased the MAO activity and 5-HT levels in the striatum. Moreover, co-treatment with naringenin reduced the expression of GFAP protein in the striatal part and significantly attenuated the neuronal cell death. The findings of the present study suggest that naringenin provides neuroprotection and mitigates neurobehavioral alterations in experimental rats.

Conclusion: The results show that co-treatment with naringenin ameliorates 3-NP-induced HD-like symptoms in rats.
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http://dx.doi.org/10.1080/1028415X.2021.1913319DOI Listing
April 2021

Brassinosteroid Signaling, Crosstalk and, Physiological Functions in Plants Under Heavy Metal Stress.

Front Plant Sci 2021 24;12:608061. Epub 2021 Mar 24.

State Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou, China.

Brassinosteroids (BRs) are group of plant steroidal hormones that modulate developmental processes and also have pivotal role in stress management. Biosynthesis of BRs takes place through established early C-6 and late C-6 oxidation pathways and the C-22 hydroxylation pathway triggered by activation of the DWF4 gene that acts on multiple intermediates. BRs are recognized at the cell surface by the receptor kinases, BRI1 and BAK1, which relay signals to the nucleus through a phosphorylation cascade involving phosphorylation of BSU1 protein and proteasomal degradation of BIN2 proteins. Inactivation of BIN2 allows BES1/BZR1 to enter the nucleus and regulate the expression of target genes. In the whole cascade of signal recognition, transduction and regulation of target genes, BRs crosstalk with other phytohormones that play significant roles. In the current era, plants are continuously exposed to abiotic stresses and heavy metal stress is one of the major stresses. The present study reveals the mechanism of these events from biosynthesis, transport and crosstalk through receptor kinases and transcriptional networks under heavy metal stress.
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http://dx.doi.org/10.3389/fpls.2021.608061DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8024700PMC
March 2021

n-Butyl Benzyl Phthalate Exposure Promotes Lesion Survival in a Murine Endometriosis Model.

Int J Environ Res Public Health 2021 03 31;18(7). Epub 2021 Mar 31.

Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.

Endometriosis is an inflammatory and estrogen-dependent gynecological disease associated with exposure to environmental endocrine disruptors. n-Butyl benzyl phthalate (BBP), a ubiquitous plasticizer, has weak estrogenic activity, and exposure to BBP is associated with endometriosis. We aimed to elucidate the immunomodulatory effect of BBP on endometriosis development. We previously established a surgery-induced endometriosis-like murine model. In the present study, we exposed those mice to BBP 10 days prior to surgery and 4 weeks after surgery at physiologically relevant doses to mimic human exposure. Chronic exposure to BBP did not promote the growth of endometriotic lesions; however, the lesion survival rate in BBP-treated mice did increase significantly compared with control mice. Multiparametric flow cytometry showed that BBP exposure did not affect the homeostasis of infiltrated immune subsets in lesions but did enhance CD44 (adhesion marker) expression on plasmacytoid dendritic cells (pDCs). Blocking CD44 interactions locally inhibited endometriotic lesion growth. Immunofluorescence results further confirmed that CD44 blocking inhibited pDC infiltration and reduced the frequency of CD44 pDCs in endometriotic tissues. BBP also disrupted the estrus cycle in these mice. This study suggests that chronic exposure to low-dose BBP may promote survival of endometriotic tissue through CD44-expressing pDCs.
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http://dx.doi.org/10.3390/ijerph18073640DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8036315PMC
March 2021

Elastin-inspired supramolecular hydrogels: a multifaceted extracellular matrix protein in biomedical engineering.

Soft Matter 2021 Mar 17;17(12):3266-3290. Epub 2021 Mar 17.

Institute of Nano Science and Technology (INST), Sector 81, Knowledge City, Mohali, 140306, Punjab, India.

The phenomenal advancement in regenerative medicines has led to the development of bioinspired materials to fabricate a biomimetic artificial extracellular matrix (ECM) to support cellular survival, proliferation, and differentiation. Researchers have diligently developed protein polymers consisting of functional sequences of amino acids evolved in nature. Nowadays, certain repetitive bioinspired polymers are treated as an alternative to synthetic polymers due to their unique properties like biodegradability, easy scale-up, biocompatibility, and non-covalent molecular associations which imparts tunable supramolecular architecture to these materials. In this direction, elastin has been identified as a potential scaffold that renders extensibility and elasticity to the tissues. Elastin-like polypeptides (ELPs) are artificial repetitive polymers that exhibit lower critical solution temperature (LCST) behavior in a particular environment than synthetic polymers and hence have gained extensive interest in the fabrication of stimuli-responsive biomaterials. This review discusses in detail the unique structural aspects of the elastin and its soluble precursor, tropoelastin. Furthermore, the versatility of elastin-like peptides is discussed through numerous examples that bolster the significance of elastin in the field of regenerative medicines such as wound care, cardiac tissue engineering, ocular disorders, bone tissue regeneration, etc. Finally, the review highlights the importance of exploring short elastin-mimetic peptides to recapitulate the structural and functional aspects of elastin for advanced healthcare applications.
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http://dx.doi.org/10.1039/d0sm02202kDOI Listing
March 2021

Tocilizumab plus standard care versus standard care in patients in India with moderate to severe COVID-19-associated cytokine release syndrome (COVINTOC): an open-label, multicentre, randomised, controlled, phase 3 trial.

Lancet Respir Med 2021 05 4;9(5):511-521. Epub 2021 Mar 4.

University Hospitals Bristol NHS Foundation Trust and Translational Health Sciences, Bristol Medical School, Bristol, UK.

Background: Global randomised controlled trials of the anti-IL-6 receptor antibody tocilizumab in patients admitted to hospital with COVID-19 have shown conflicting results but potential decreases in time to discharge and burden on intensive care. Tocilizumab reduced progression to mechanical ventilation and death in a trial population enriched for racial and ethnic minorities. We aimed to investigate whether tocilizumab treatment could prevent COVID-19 progression in the first multicentre randomised controlled trial of tocilizumab done entirely in a lower-middle-income country.

Methods: COVINTOC is an open-label, multicentre, randomised, controlled, phase 3 trial done at 12 public and private hospitals across India. Adults (aged ≥18 years) admitted to hospital with moderate to severe COVID-19 (Indian Ministry of Health grading) confirmed by positive SARS-CoV-2 PCR result were randomly assigned (1:1 block randomisation) to receive tocilizumab 6 mg/kg plus standard care (the tocilizumab group) or standard care alone (the standard care group). The primary endpoint was progression of COVID-19 (from moderate to severe or from severe to death) up to day 14 in the modified intention-to-treat population of all participants who had at least one post-baseline assessment for the primary endpoint. Safety was assessed in all randomly assigned patients. The trial is completed and registered with the Clinical Trials Registry India (CTRI/2020/05/025369).

Findings: 180 patients were recruited between May 30, 2020, and Aug 31, 2020, and randomly assigned to the tocilizumab group (n=90) or the standard care group (n=90). One patient randomly assigned to the standard care group inadvertently received tocilizumab at baseline and was included in the tocilizumab group for all analyses. One patient randomly assigned to the standard care group withdrew consent after the baseline visit and did not receive any study medication and was not included in the modified intention-to-treat population but was still included in safety analyses. 75 (82%) of 91 in the tocilizumab group and 68 (76%) of 89 in the standard care group completed 28 days of follow-up. Progression of COVID-19 up to day 14 occurred in eight (9%) of 91 patients in the tocilizumab group and 11 (13%) of 88 in the standard care group (difference -3·71 [95% CI -18·23 to 11·19]; p=0·42). 33 (36%) of 91 patients in the tocilizumab group and 22 (25%) of 89 patients in the standard care group had adverse events; 18 (20%) and 15 (17%) had serious adverse events. The most common adverse event was acute respiratory distress syndrome, reported in seven (8%) patients in each group. Grade 3 adverse events were reported in two (2%) patients in the tocilizumab group and five (6%) patients in the standard care group. There were no grade 4 adverse events. Serious adverse events were reported in 18 (20%) patients in the tocilizumab group and 15 (17%) in the standard care group; 13 (14%) and 15 (17%) patients died during the study.

Interpretation: Routine use of tocilizumab in patients admitted to hospital with moderate to severe COVID-19 is not supported. However, post-hoc evidence from this study suggests tocilizumab might still be effective in patients with severe COVID-19 and so should be investigated further in future studies.

Funding: Medanta Institute of Education and Research, Roche India, Cipla India, and Action COVID-19 India.
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http://dx.doi.org/10.1016/S2213-2600(21)00081-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8078880PMC
May 2021

A Global eDelphi Exercise to Identify Core Domains and Domain Items for the Development of a Global Registry of Alopecia Areata Disease Severity and Treatment Safety (GRASS).

JAMA Dermatol 2021 Apr;157(4):1-11

Clinical Medicine, Trinity College Dublin, Dublin, Ireland.

Importance: A recent expert consensus exercise emphasized the importance of developing a global network of patient registries for alopecia areata to redress the paucity of comparable, real-world data regarding the effectiveness and safety of existing and emerging therapies for alopecia areata.

Objective: To generate core domains and domain items for a global network of alopecia areata patient registries.

Evidence Review: Sixty-six participants, representing physicians, patient organizations, scientists, the pharmaceutical industry, and pharmacoeconomic experts, participated in a 3-round eDelphi process, culminating in a face-to-face meeting at the World Congress of Dermatology, Milan, Italy, June 14, 2019.

Findings: Ninety-two core data items, across 25 domains, achieved consensus agreement. Twenty further noncore items were retained to facilitate data harmonization in centers that wish to record them. Broad representation across multiple stakeholder groups was sought; however, the opinion of physicians was overrepresented.

Conclusions And Relevance: This study identifies the domains and domain items required to develop a global network of alopecia areata registries. These domains will facilitate a standardized approach that will enable the recording of a comprehensive, comparable data set required to oversee the introduction of new therapies and harness real-world evidence from existing therapies at a time when the alopecia areata treatment paradigm is being radically and positively disrupted. Reuse of similar, existing frameworks in atopic dermatitis, produced by the Treatment of Atopic Eczema (TREAT) Registry Taskforce, increases the potential to reuse existing resources, creates opportunities for comparison of data across dermatology subspecialty disease areas, and supports the concept of data harmonization.
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http://dx.doi.org/10.1001/jamadermatol.2020.5839DOI Listing
April 2021

Efficiency of bacteria and bacterial assisted phytoremediation of heavy metals: An update.

Authors:
Pooja Sharma

Bioresour Technol 2021 May 14;328:124835. Epub 2021 Feb 14.

Department of Environmental Microbiology, School for Environmental Sciences, Babasaheb Bhimrao Ambedkar Central University, Lucknow 226 025, Uttar Pradesh, India.

The aim of this review to address the plant-associated bacteria to enhance the phytoremediation efficiency of the heavy metals from polluted sites and it is also highlighted advances for the application in wastewater treatment. Plant-associated bacteria have potential to encourage the plant growth and resistance under stress conditions. Such bacteria could enhance plant growth by controlling growth hormone, nutrition security, producing siderophore, secondary metabolites, and improving the antioxidant enzymes system. This review also explores the concepts and applications of bacteria assisted phytoremediation, addressing aspects that affect phytoremediation and pathways for restoration. Significant review issues relating to production and application of bacteria for improvement of bioremediation were established and presented for possible future research. Bacteria assisted phytoremediation is cost-effective strategy and metal sequestration mechanism that hold high metal biosorption capacities. This also takes into consideration the current state of technology implementations and proposals for prospective clean-up studies.
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http://dx.doi.org/10.1016/j.biortech.2021.124835DOI Listing
May 2021

Role of microbial community and metal-binding proteins in phytoremediation of heavy metals from industrial wastewater.

Bioresour Technol 2021 Apr 22;326:124750. Epub 2021 Jan 22.

CSIR-National Environmental and Engineering Research Institute (CSIR-NEERI), Nehru Marg, Nagpur 440 020, India. Electronic address:

This review illustrated the role of metal-binding proteins (MBPs) and microbial interaction in assisting the phytoremediation of industrial wastewater polluted with heavy metals. MBPs are used to increase the accumulation and tolerance of metals by microorganisms via binding protein synthesis. Microbes have various protection mechanisms to heavy metals stress like compartmentalization, exclusion, complexity rendering, and the synthesis of binding proteins. MBPs include phytochelatins, metallothioneins, Cd-binding peptides (CdBPs), cysteines (gcgcpcgcg) (CP), and histidines (ghhphg) (HP). In comparison with other physico-chemical methods, phytoremediation is an eco-friendly and safe method for the society. The present review concentrated on the efficiency of phytoremediation strategies for the use of MBPs and microbe-assisted approaches.
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http://dx.doi.org/10.1016/j.biortech.2021.124750DOI Listing
April 2021

Metagenomic analysis for profiling of microbial communities and tolerance in metal-polluted pulp and paper industry wastewater.

Bioresour Technol 2021 Mar 8;324:124681. Epub 2021 Jan 8.

Department of Environmental Microbiology, School for Environmental Sciences, Babasaheb Bhimrao Ambedkar University (A Central University), Lucknow 226 025, Uttar Pradesh, India. Electronic address:

This work aimed to study the profiling and efficiency of microbial communities and their abundance in the pulp and paper industry wastewater, which contained toxic metals, high biological oxygen demands, chemical oxygen demand, and ions contents. Sequence alignment of the 16S rRNA V3-V4 variable region zone with the Illumina MiSeq framework revealed 25356 operating taxonomical units (OTUs) derived from the wastewater sample. The major phyla identified in wastewater were Proteobacteria, Bacteroidetes, Firmicutes, Chloroflexi, Actinobacteria, Spirochetes, Patesibacteria, Acidobacteria, and others including unknown microbes. The study showed the function of microbial communities essential for the oxidation and detoxifying of complex contaminants and design of effective remediation techniques for the re-use of polluted wastewater. Findings demonstrated that the ability of different classes of microbes to adapt and survive in metal-polluted wastewater irrespective of their relative distribution, as well as further attention can be provided to its use in the bioremediation process.
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http://dx.doi.org/10.1016/j.biortech.2021.124681DOI Listing
March 2021

Residual pollutants in treated pulp paper mill wastewater and their phytotoxicity and cytotoxicity in Allium cepa.

Environ Geochem Health 2021 May 5;43(5):2143-2164. Epub 2021 Jan 5.

Department of Environmental Microbiology, School for Environmental Sciences, Babasaheb Bhimrao Ambedkar Central University, Vidya Vihar, Rae Bareli Road, Lucknow, Uttar Pradesh, 226025, India.

Discharged pulp and paper mill wastewater (PPMW) were collected near M/s K. R. pulp and papers Limited, Shahjahanpur, India. Chemical analysis of the wastewater showed high BOD (3653-4180 mg L) and COD (17,890-19100 mg L) values from two different sampling sites. The levels of total phenol were in the range of 389-432 mg L; nitrogen (125-234 mg L), sulfate (1926-2098 mg L), chloride (3.12-5.43 mg L) and lignin (38,950-39,000 mg L) along with various heavy metals (Fe, 87-79; Zn, 34-22; Cu, 3.28-2.57; Cd, 1.90-0.36; Ni, 6-5, and Pb, 41.23-36.54 mg L) were above the permissible limits recommended by the CPCB and the USEPA. The BOD/COD ratio was  < 0.2 which indicated very low biodegradability of the organic matters present in the effluent. The organometallic complex generated from the pulp and paper industry persists in the environment and might be toxic to aquatic organisms. The organic polymers, lignin, metals and ions present in the PPMW were characterized using SEM, EDAX, FTIR, and UV-VIS spectroscopy. The major pollutants detected in the discharged PPMW included nonacosane, heptacosane, octadecanoic acid, hexadecane, and 6-benzamide- 3- [2- [1-(phenylmethyl)-4-piperidinyl] ethyl]-1, 2-benzisoxazole, as well as a group of plant fatty acids classified as EDCs, and mutagenic pollutants. The cytotoxic and androgenic properties of these complex organics were examined. The seed germination test with Phaseolus mungo and cytotoxicity test with Allium cepa showed that at > 20% concentration of PPMW, α-amylase production was inhibited and chromosomal segregation at metaphase and anaphase during cell division was disturbed, which resulted in c-mitosis, sticky chromosomes, and laggard chromosomes. In addition, SEM of the root of A. cepa showed fissures and fractured tissues of the root cap, probably due to the inhibition of auxins that were responsible for root cap formation. The findings indicated A. cepa as a good test model for examining the DNA damage and cytotoxicity by PPMW, and the discharged effluent should be treated at a tertiary stage for environmental protection.
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http://dx.doi.org/10.1007/s10653-020-00730-zDOI Listing
May 2021

Keratin 19 maintains E-cadherin localization at the cell surface and stabilizes cell-cell adhesion of MCF7 cells.

Cell Adh Migr 2021 12;15(1):1-17

Department of Biology, The Catholic University of America , Washington, District of Columbia, USA.

A cytoskeletal protein keratin 19 (K19) is highly expressed in breast cancer but its effects on breast cancer cell mechanics are unclear. In MCF7 cells where K19 expression is ablated,  knockout cells, suggesting that E-cadherin internalization contributed to defective adhesion. Ultimately, while K19 inhibited cell migration and invasion, it was required for cells to form colonies in suspension. Our results suggest that K19 stabilizes E-cadherin complexes at the cell membrane to maintain cell-cell adhesion which inhibits cell invasiveness but provides growth and survival advantages for circulating tumor cells.
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http://dx.doi.org/10.1080/19336918.2020.1868694DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7801129PMC
December 2021

Casal's necklace.

Natl Med J India 2019 Nov-Dec;32(6):378

Department of Dermatology and Venereology, All India Institute of Medical Sciences, New Delhi, India.

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http://dx.doi.org/10.4103/0970-258X.303626DOI Listing
December 2020

Evaluation of the Performance of CinTec® PLUS in SurePathTM Liquid-Based Cervico-Vaginal Samples.

Turk Patoloji Derg 2021 ;37(1):32-38

Department of Cytology and Gynecological Pathology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.

Objective: Cervical cytology and Human papillomavirus (HPV) testing are effective screening techniques but both have limitations. A few recent studies in the literature have highlighted the role of co-expression of p16INK4a and Ki-67 for cervical cancer screening. The present study was undertaken to evaluate the diagnostic performance of the CINtec® PLUS kit (dual immunostaining for p16 and Ki-67) in SurePathTM liquid-based (LBC) cervico-vaginal samples.

Materials And Methods: This was a prospective study performed on 52 cervico-vaginal SurePath™ LBC samples reported as having squamous epithelial cell abnormality (ECA). All the samples were stained using CINtec® PLUS kits. Additionally, HPV-DNA testing was also done and the results were compared.

Results: The age range was 34-74 years. ECA included 18 (34.6%) cases of ASC-US, 9 (17.3%) cases of low-grade squamous intraepithelial lesion (LSIL), 11 (21.2%) cases of high-grade squamous intraepithelial lesion (HSIL), and 14 (26.9%) cases of squamous cell carcinoma (SCC). Cervical biopsies were available in 19 (36.5%) cases. A total of 34/52 (65.4%) cases were positive for HPV-DNA (5/18-ASC-US; 6/9-LSIL; 10/11-HSIL; 13/14-SCC). The CINtec® PLUS test was positive in 41/52 (78.8%) cases (11/18-ASC-US; 6/9-LSIL; 11/11-HSIL; 13/14-SCC). On comparing CINtec® PLUS positivity (78.8%) with HPV positivity (65.4%), dual positivity was seen in 3/18 cases of ASC-US, 6/9 cases of LSIL, 10/11 cases of HSIL, and 12/14 cases of SCC. One case each of HSIL and SCC was negative on the HPV test and was positive on CINtec® PLUS.

Conclusions: CINtec® PLUS test helps to improve the detection of pre-cancerous cervical lesions as compared to cervical cytology or HPV testing alone and hence can serve as a potentially useful diagnostic and triage tool, especially for indeterminate cases.
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http://dx.doi.org/10.5146/tjpath.2020.01505DOI Listing
January 2021

In vitro, in vivo and in silico Antihyperglycemic Activity of Some Semi-synthetic Phytol Derivatives.

Med Chem 2020 Dec 16. Epub 2020 Dec 16.

Medicinal Chemistry Department, Metabolic and Structural Biology Department; CSIR-Central Institute of Medicinal and Aromatic Plants, P.O.- CIMAP, Lucknow-226015. India.

Background: Due to the prevalence of type-2 diabetes across the globe, there is unmet need to explore new molecular targets for the development of cost-effective and safer antihyperglycemic agents.

Objective: Structural modification of phytol and evaluation of in vitro, in vivo and in silico antihyperglycemic activity of derivatives establishing the preliminary structure activity relationship (SAR).

Methods: The semi-synthetic derivatives of phytol were prepared following previously described methods. The antihyperglycemic potential was measured in vitro in terms of increase in 2-deoxyglucose (2-DG) uptake by L-6 rat skeletal muscle cells as well as in vivo in sucrose-loaded (SLM) and streptozotocin (STZ)-induced diabetic rat models. The blood glucose profile was measured at 30, 60, 90, 120, 180, 240, 300 and 1440 min post administration of sucrose in rats. The in silico docking was performed on peroxisome proliferator-activated receptor gamma (PPARγ) as anti-diabetic target along with absorption, distribution, metabolism, excretion and toxicity (ADMET) studies.

Results: Nine semi-synthetic ester derivatives: acetyl (1), lauroyl (2), palmitoyl (3), pivaloyl (4), trans-crotonyl (5), benzoyl (6), m-anisoyl (7), 3,4,5-trimethoxy benzoyl (8) cinnamoyl (9) along with bromo derivative (10) of phytol were prepared. The derivatives 9, 8 and 2 caused 4.5, 3.2 and 2.7 times more in vitro uptake of 2-DG respectively than rosiglitazone (ROSI). The derivatives showed significant improvement on oral glucose tolerance both in SLM (29.6-21%) as well as STZ-induced diabetic (30.8-19.0%) rats. The in silico ADMET, docking studies showed non-toxicity and high binding affinity with PPARγ.

Conclusion: The potent antihyperglycemic activity with favorable pharmacokinetics supports phytol derivatives as suitable antidiabetic lead.
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http://dx.doi.org/10.2174/1573406417666201216124018DOI Listing
December 2020

Synthesis of Dendritic Glycoclusters and Their Applications for Supramolecular Gelation and Catalysis.

J Org Chem 2020 12 10;85(24):16136-16156. Epub 2020 Dec 10.

Department of Chemistry and Biochemistry, Old Dominion University, Norfolk, Virginia 23529, United States.

Glycoclusters with three, four, and six arms of glycosyl triazoles were designed, synthesized, and characterized. The self-assembling properties of these molecules and their catalytic activity as ligands in copper-catalyzed azide and alkyne cycloaddition (CuAAC) reactions were studied. The compounds with a lower number of branches exhibit excellent gelation properties and can function as supramolecular gelators. The resulting gels were characterized using optical microcopy and atomic force microscopy. The glycoconjugates containing six branches showed significant catalytic activity for copper sulfate mediated cycloaddition reactions. In aqueous solutions, 1 mol % of glycoclusters to substrates was efficient at accelerating these reactions. Several trimeric compounds were found to be capable of forming co-gels with the catalytically active hexameric compounds. Using the organogels formed by the glycoconjugates as supramolecular catalysts, efficient catalysis was demonstrated for several CuAAC reactions. The metallogels with CuSO were also prepared as gel columns, which can be reused for the cycloaddition reactions several times. These include the preparation of a few glycosyl triazoles and aryl triazoles and isoxazoles. We expect that these sugar-based soft biomaterials will have applications beyond supramolecular catalysis for copper-catalyzed cycloaddition reactions. They may also be useful as ligands or gel matrixes for other metal-ion catalyzed organic reactions.
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http://dx.doi.org/10.1021/acs.joc.0c01978DOI Listing
December 2020

Determining crucial genes associated with COVID-19 based on COPD Findings.

Comput Biol Med 2021 01 21;128:104126. Epub 2020 Nov 21.

Department of CSE,Tezpur University, Assam, 784028, India.

Genes act in groups known as gene modules, which accomplish different cellular functions in the body. The modular nature of gene networks was used in this study to detect functionally enriched modules in samples obtained from COPD patients. We analyzed modules extracted from COPD samples and identified crucial genes associated with the disease COVID-19. We also extracted modules from a COVID-19 dataset and analyzed a suspected set of genes that may be associated with this deadly disease. We used information available for two other viruses that cause SARS and MERS because their physiology is similar to that of the COVID-19 virus. We report several crucial genes associated with COVID-19: RPA2, POLD4, MAPK8, IRF7, JUN, NFKB1, NFKBIA, CD40LG, FASLG, ICAM1, LIFR, STAT2 and CCR1. Most of these genes are related to the immune system and respiratory organs, which emphasizes the fact that COPD weakens this system and makes patients more susceptible to developing severe COVID-19.
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http://dx.doi.org/10.1016/j.compbiomed.2020.104126DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7680043PMC
January 2021

Newly isolated Bacillus sp. PS-6 assisted phytoremediation of heavy metals using Phragmites communis: Potential application in wastewater treatment.

Bioresour Technol 2021 Jan 7;320(Pt B):124353. Epub 2020 Nov 7.

Department of Environmental Microbiology, School for Environmental Sciences, Babasaheb Bhimrao Ambedkar University, Lucknow (A Central University) 226025, Uttar Pradesh, India. Electronic address:

This work aimed to study Bacillus sp. PS-6 assisted phytoremediation of metals from pulp and paper industry wastewater as a novel green technique for the removal of metals of wastewater. Results revealed that heavy metal (mg L) contents in wastewater were reduced after in-situ phytoremediation for Fe, Cu, Zn, Cd, Mn, Ni, Pb, and As. Phragmites communis showed higher potential for the enrichment of Fe, Cu, Zn, Cd, Mn, Ni, Pb, and As in its rhizomes, roots, and shoots compared to leaves. The strain produced indole acetic acid, siderophores, and hydrolytic and ligninolytic enzymes, and resulted in nutrients solubilization. Results offer potential basis for the removal of metals from pulp and paper industry wastewater at large scale and prevention of pollution.
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http://dx.doi.org/10.1016/j.biortech.2020.124353DOI Listing
January 2021

Integrated genomic view of SARS-CoV-2 in India.

Wellcome Open Res 2020 3;5:184. Epub 2020 Aug 3.

Biotechnology Division, National Centre for Disease Control, Delhi, Delhi, 110054, India.

India first detected SARS-CoV-2, causal agent of COVID-19 in late January 2020, imported from Wuhan, China. From March 2020 onwards, the importation of cases from countries in the rest of the world followed by seeding of local transmission triggered further outbreaks in India. We used ARTIC protocol-based tiling amplicon sequencing of SARS-CoV-2 (n=104) from different states of India using a combination of MinION and MinIT sequencing from Oxford Nanopore Technology to understand how introduction and local transmission occurred. The analyses revealed multiple introductions of SARS-CoV-2 genomes, including the A2a cluster from Europe and the USA, A3 cluster from Middle East and A4 cluster (haplotype redefined) from Southeast Asia (Indonesia, Thailand and Malaysia) and Central Asia (Kyrgyzstan). The local transmission and persistence of genomes A4, A2a and A3 was also observed in the studied locations. The most prevalent genomes with patterns of variance (confined in a cluster) remain unclassified, and are here proposed as A4-clade based on its divergence within the A cluster. The viral haplotypes may link their persistence to geo-climatic conditions and host response. Multipronged strategies including molecular surveillance based on real-time viral genomic data is of paramount importance for a timely management of the pandemic.
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http://dx.doi.org/10.12688/wellcomeopenres.16119.1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7506191PMC
August 2020

Highly efficient phytoremediation potential of metal and metalloids from the pulp paper industry waste employing Eclipta alba (L) and Alternanthera philoxeroide (L): Biosorption and pollution reduction.

Bioresour Technol 2021 Jan 21;319:124147. Epub 2020 Sep 21.

Department of Environmental Microbiology, School for Environmental Sciences, Babasaheb Bhimrao Ambedkar Central University, Lucknow 226 025, Uttar Pradesh, India. Electronic address:

The aims of the study was the evaluation of phytoremediation potential by Eclipta alba (L) and Alternanthera philoxeroide (L) of pulp and paper mill waste after secondary treatment which a source of aquatic and soil pollution due to huge discharge of organometallic compounds per tone of paper production. The result revealed 50% reduction of pollution parameters after in-situ phytoremediation. The comparative analysis of metal and metalloids showed the highest accumulation of Fe (2251.24 ± 64.74) in both plants. The antioxidant activity, chlorophyll and carotenoid content were increased in E. alba (L.) and A. philoxeroide (L.) respectively. From the results, it was concluded that E. alba (L.) and A. philoxeroide (L.) could be effectively used for the removal of metals and metalloids from effluent and sludge of pulp and paper mill waste that may help to reduce adverse health effects of metal accumulation in humans and animals via their food chain.
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http://dx.doi.org/10.1016/j.biortech.2020.124147DOI Listing
January 2021

Accessing Highly Tunable Nanostructured Hydrogels in a Short Ionic Complementary Peptide Sequence via pH Trigger.

Langmuir 2020 10 9;36(41):12107-12120. Epub 2020 Oct 9.

Habitat Centre, Institute of Nano Science and Technology, Sector 64, Phase 10, Mohali, Punjab 160062, India.

Creating diverse nanostructures from a single gelator through modulating the self-assembly pathway has been gaining much attention in recent years. To this direction, we are exploring the effect of modulation of pH as a potential self-assembly pathway in governing the physicochemical properties of the final gel phase material. In this context, we used a classical nongelator with the ionic complementary sequence FEFK, which was rationally conjugated to an aromatic group naphthoxyacetic acid (Nap) at the N-terminal end to tune its gelation behavior. Interestingly, the presence of oppositely charged amino acids in the peptide amphiphile resulted in pH-responsive behavior, leading to the formation of hydrogels over a wide pH range (2.0-12.0); however, their structures differ significantly at the nanoscale. Thus, by simply manipulating the overall charge over the exposed surface of the peptide amphiphiles as a function of pH, we were able to access diverse self-assembled nanostructures within a single gelator domain. The charged state of the gelator at the extreme pH (2.0, 12.0) led to a thinner fiber formation, in contrast to the thicker fibers observed near the physiological pH owing to charge neutralization, thus promoting the lateral association. Such variation in molecular packing was found to be further reflected in the variable mechanical strengths of the peptide hydrogels obtained at different pH values. Moreover, the gelation of the peptide at physiological pH offers an additional advantage to explore this hydrogel as a cell culture scaffold. We anticipate that our study on controlling the self-assembly pathway of the ionic complementary peptide amphiphile can be an elegant approach to access diverse self-assembled materials, which can expand the zone of its applicability as a stimuli-responsive biomaterial.
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http://dx.doi.org/10.1021/acs.langmuir.0c01472DOI Listing
October 2020

The Alopecia Areata Consensus of Experts (ACE) study part II: Results of an international expert opinion on diagnosis and laboratory evaluation for alopecia areata.

J Am Acad Dermatol 2021 Jun 12;84(6):1594-1601. Epub 2020 Sep 12.

Duke Dermatology Clinic-Clinic 3K, Durham, North Carolina.

Background: We previously reported the Alopecia Areata Consensus of Experts study, which presented results of an international expert opinion on treatments for alopecia areata.

Objective: To report the results of the Alopecia Areata Consensus of Experts international expert opinion on diagnosis and laboratory evaluation for alopecia areata.

Methods: Fifty hair experts from 5 continents were invited to participate in a 3-round Delphi process. Consensus threshold was set at greater than or equal to 66%.

Results: Of 148 questions, expert consensus was achieved in 82 (55%). Round 1 consensus was achieved in 10 of 148 questions (7%). Round 2 achieved consensus in 47 of 77 questions (61%). The final face-to-face achieved consensus in 25 of 32 questions (78%). Consensus was greatest for laboratory evaluation (12 of 14 questions [86%]), followed by diagnosis (11 of 14 questions [79%]) of alopecia areata. Overall, etiopathogenesis achieved the least category consensus (31 of 68 questions [46%]).

Limitations: The study had low representation from Africa, South America, and Asia.

Conclusion: There is expert consensus on aspects of epidemiology, etiopathogenesis, clinical features, diagnosis, laboratory evaluation, and prognostic indicators of alopecia areata. The study also highlights areas where future clinical research could be directed to address unresolved hypotheses in alopecia areata patient care.
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http://dx.doi.org/10.1016/j.jaad.2020.09.028DOI Listing
June 2021

Frequency spectrum of rare and clinically relevant markers in multiethnic Indian populations (ClinIndb): A resource for genomic medicine in India.

Hum Mutat 2020 Nov 9;41(11):1833-1847. Epub 2020 Sep 9.

Genomics and Molecular Medicine, CSIR-Institute of Genomics and Integrative Biology, New Delhi, India.

There have been concerted efforts toward cataloging rare and deleterious variants in different world populations using high-throughput genotyping and sequencing-based methods. The Indian population is underrepresented or its information with respect to clinically relevant variants is sparse in public data sets. The aim of this study was to estimate the burden of monogenic disease-causing variants in Indian populations. Toward this, we have assessed the frequency profile of monogenic phenotype-associated ClinVar variants. The study utilized a genotype data set (global screening array, Illumina) from 2795 individuals (multiple in-house genomics cohorts) representing diverse ethnic and geographically distinct Indian populations. Of the analyzed variants from Global Screening Array, ~9% were found to be informative and were either not known earlier or underrepresented in public databases in terms of their frequencies. These variants were linked to disorders, namely inborn errors of metabolism, monogenic diabetes, hereditary cancers, and various other hereditary conditions. We have also shown that our study cohort is genetically a better representative of the Indian population than its representation in the 1000 Genome Project (South Asians). We have created a database, ClinIndb, linked to the Leiden Open Variation Database, to help clinicians and researchers in diagnosis, counseling, and development of appropriate genetic screening tools relevant to the Indian populations and Indians living abroad.
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http://dx.doi.org/10.1002/humu.24102DOI Listing
November 2020

Potent Inhibition of Necroptosis by Simultaneously Targeting Multiple Effectors of the Pathway.

ACS Chem Biol 2020 10 22;15(10):2702-2713. Epub 2020 Sep 22.

The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC 3052, Australia.

Necroptosis is an inflammatory form of programmed cell death that has been implicated in various human diseases. Compound is a more potent analogue of the published compound and inhibits necroptosis in human and murine cells at nanomolar concentrations. Several target engagement strategies were employed, including cellular thermal shift assays (CETSA) and diazirine-mediated photoaffinity labeling via a bifunctional photoaffinity probe derived from compound . These target engagement studies demonstrate that compound binds to all three necroptotic effector proteins (mixed lineage kinase domain-like protein (MLKL), receptor-interacting serine/threonine protein kinase 1 (RIPK1) and receptor-interacting serine/threonine protein kinase 3 (RIPK3)) at different levels and in cells. Compound also shows efficacy in a murine model of systemic inflammatory response syndrome (SIRS).
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http://dx.doi.org/10.1021/acschembio.0c00482DOI Listing
October 2020

Structural and functional characterization of C0021158, a high-affinity monoclonal antibody that inhibits Arginase 2 function via a novel non-competitive mechanism of action.

MAbs 2020 Jan-Dec;12(1):1801230

Cancer Research UK AstraZeneca Antibody Alliance Laboratory , Cambridge, UK.

Arginase 2 (ARG2) is a binuclear manganese metalloenzyme that catalyzes the hydrolysis of L-arginine. The dysregulated expression of ARG2 within specific tumor microenvironments generates an immunosuppressive niche that effectively renders the tumor 'invisible' to the host's immune system. Increased ARG2 expression leads to a concomitant depletion of local L-arginine levels, which in turn leads to suppression of anti-tumor T-cell-mediated immune responses. Here we describe the isolation and characterization of a high affinity antibody (C0021158) that inhibits ARG2 enzymatic function completely, effectively restoring T-cell proliferation . Enzyme kinetic studies confirmed that C0021158 exhibits a noncompetitive mechanism of action, inhibiting ARG2 independently of L-arginine concentrations. To elucidate C0021158's inhibitory mechanism at a structural level, the co-crystal structure of the Fab in complex with trimeric ARG2 was solved. C0021158's epitope was consequently mapped to an area some distance from the enzyme's substrate binding cleft, indicating an allosteric mechanism was being employed. Following C0021158 binding, distinct regions of ARG2 undergo major conformational changes. Notably, the backbone structure of a surface-exposed loop is completely rearranged, leading to the formation of a new short helix structure at the Fab-ARG2 interface. Moreover, this large-scale structural remodeling at ARG2's epitope translates into more subtle changes within the enzyme's active site. An arginine residue at position 39 is reoriented inwards, sterically impeding the binding of L-arginine. Arg39 is also predicted to alter the p of a key catalytic histidine residue at position 160, further attenuating ARG2's enzymatic function. molecular docking simulations predict that L-arginine is unable to bind effectively when antibody is bound, a prediction supported by isothermal calorimetry experiments using an L-arginine mimetic. Specifically, targeting ARG2 in the tumor microenvironment through the application of C0021158, potentially in combination with standard chemotherapy regimens or alternate immunotherapies, represents a potential new strategy to target immune cold tumors.
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http://dx.doi.org/10.1080/19420862.2020.1801230DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7531564PMC
September 2020

"Metal-Free" Fluorescent Supramolecular Assemblies for Distinct Detection of Organophosphate/Organochlorine Pesticides.

ACS Omega 2020 Aug 31;5(31):19654-19660. Epub 2020 Jul 31.

Department of Chemistry, UGC Sponsored Centre of Advanced Studies-II, Guru Nanak Dev University, Amritsar 143005, Punjab, India.

The "metal-free", easy-to-prepare fluorescent supramolecular assemblies based on anthracene/perylene bisamide (PBI) derivatives have been developed for the distinct detection of organophosphate (CPF) and organochlorine (DCN) pesticides in aqueous media. The supramolecular assemblies of anthracene derivative show rapid and highly selective "on-on" response toward organophosphate (CPF), which is attributed to the formation of CPF-induced formation of "closely packed" assemblies. A detection limit in the nanomolar range is observed for CPF. On the other hand, the inner filter effect is proposed as the mechanism for the "on-off" detection of DCN using supramolecular assemblies of the anthracene derivative. This is the first report on the development of fluorescent materials having the potential to differentiate between organophosphate and organochlorine pesticides. The assemblies of anthracene derivative also act as "enzyme mimic" as organophosphate pesticide show a preferential affinity for assemblies of derivative over acetylcholinesterase enzyme. Further, the real-time applications of supramolecular assemblies have also been explored for the detection of CPF and DCN in spiked water and in agricultural products such as grapes and apples.
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http://dx.doi.org/10.1021/acsomega.0c02315DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7424749PMC
August 2020

Phytoremediation potential of heavy metal accumulator plants for waste management in the pulp and paper industry.

Heliyon 2020 Jul 28;6(7):e04559. Epub 2020 Jul 28.

Department of Environmental Microbiology, School for Environmental Sciences, Babasaheb Bhimrao Ambedkar Central University, Vidya Vihar, Raebareli Road, Lucknow, Uttar Pradesh 226025, India.

The present manuscript has focused on the heavy metal; accumulation potential by common native plants i.e. L., , and growing on the disposed of pulp and paper mill effluent sludge. The sludge showed the abundance of benzene propanoic acid tert- butyldimethylsilyl ester, Octadecanoic acid, TMS, Hexadecanoic acid, TMS, cinnamic acid-α-phenyl-TMS ester, β-sitosterol TMS, 4-mercaptobenzoic acid as residual complex organic compounds along with heavy metals Fe (98.30 mg/L), Zn (51.00 mg/L), Cu (3.21 mg/L), Cd (9.11 mg/L), Mn (18.27 mg/L), Ni (5.21 mg/L), (Hg 0.014 mg/L) which were above the prescribed limit of environmental standard. The complexation of organic compounds with heavy metal restricts the bioavailability of metals to plants. But the metal analysis in various parts of the plant showed a significant amount of metal accumulation. Further, histological observations of root tissue through TEM showed apparent deposition of metal granules near the cell wall and vacuole as adoption features of plants. But the variable concentration of metal accumulation in different parts by various plants indicated the variable potential of tested plants with various metals. This also indicated their metal bio-availability and movement to plant tissue. Further, their bioconcentration factor (BCF) and translocation factor (TF) > 1.0 indicated the hyperaccumulation tendency of plants Mn was accumulated maximum in leaves (69.38 mg/kg) followed by Cu (25.75 mg/kg ), As (23.20 mg/kg ), Fe (20.90 mg/kg ) and Pb was maximum accumulated (22.41 mg/kg ) in leaves. The result revealed that arsenic has been accumulated in higher amount root, shoot and leaves of all tested plants. The metal accumulator plants showed phytoremediation potential also by reducing various pollution parameters after growth on sludge. These potential plants may be used as biotechnological tools for the eco-restoration of polluted sites.
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http://dx.doi.org/10.1016/j.heliyon.2020.e04559DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7393463PMC
July 2020