Biology subjects

Rasool, S.

Publications and source records attributed to Rasool, S..

9 recordsLinked to original sources

bioRxiv2025

Phytochemicals and Antioxidant Activity of Leaf Extract and Callus Cultures of Cinnamomum camphora L.

Cinnamomum camphora L. is highly significant landscape tree known for its medicinal values and presence of secondary metabolites that have antioxidant, antimicrobial, anticancer, anti-inflammatory effects and widely utilized in pharmaceutical and cosmetic industry. The present work was aimed at comparative analysis of phytochemicals (phenolic and flavonoid) and antioxidant activities of calli and leaf extract of field grown camphor plant. To get in vitro germplasm, callus formation and direct shoot initiation was carried out and it was observed that MS medium supplemented with 0.4 {micro}M thidiazuron (TDZ) + 2.26 {micro}M 2, 4-dichlorophenoxyacetic acid (2, 4-D) proved best for shoot initiation from nodal explant. MS medium fortified with various plant growth regulators was used for callus formation and best callus induction response (100%) from nodal and leaf explants was observed on 2.26 {micro}M 2, 4-dichlorophenoxyacetic acid (2, 4-D) + 8.87 {micro}M 6-Benzylaminopurine (BAP). Callus was successfully sub-cultured and this in vitro proliferated callus extract and fresh leaf extract of field grown plant were used for comparative study of phytochemicals. The antioxidant enzyme activity of peroxidase (31.12 UmL-1 of enzyme), superoxide dismutase (35.24 Umg-1 of protein), and catalase (58.6 UmL-1 of enzyme) was higher in callus as compared to the leaf extract while glutathione peroxidase activity (0.552 Umg-1) was comparatively higher in leaf extract rather than callus cultures. Higher phenolic contents (1.106 mg GAE g-1 of FW) were recorded in callus cultures, however flavonoid contents (7.87 mg CatE g-1 of FW) were higher in leaf extract. This investigation showed that in vitro conditions and the use of plant growth regulators in various combinations might be work as elicitors to enhance the phytochemicals and antioxidant enzymes in callus culture as compared to the leaf extract.

pharmacology and toxicology↗

bioRxiv2024

Apparent digestibility and nutritional performance of bakery waste produced in Punjab Pakistan for Labeo rohita fingerlings

This study aimed to examine the impact of bakery waste supplementation on the nutrient digestibility and performance metrics of rohu (Labeo rohita) juveniles. Fish were fed with seven different diets containing bakery waste products (biscuits, cream cake, bread, and dry rusk) in combination with rice polish and wheat flour. Chromic oxide (1%) was added to the diet as an inert marker to measure nutrient digestibility. The results indicated a significant difference (p < 0.05) in the digestibility of crude protein, lipid, gross energy, and dry matter among the dietary groups. Fish fed with rice polish had the lowest apparent digestibility value of protein (16.13 {+/-} 1.86), whereas fish fed wheat flour had the highest (77.70 {+/-} 0.05). Moreover, it was shown that fish fed rusk waste had better growth. It has been concluded that bakery wastes could be used as a cheaper fish feed ingredient source for the manufacturing of cost-effective feed for L. rohita. This investigation was discussed within the context of providing cost effective ingredients for sustainable aquaculture production in Pakistan.

zoology↗

bioRxiv2023

In-silico analysis of Myocardial Infarction-related missense SNPs to identify novel biomarkers to predict susceptibility.

Myocardial Infarction (MI), commonly known as a heart attack, stands as a formidable global health challenge, responsible for a substantial burden of morbidity and mortality. This study embarked on a comprehensive exploration of the genetic underpinnings of MI, recognizing the pivotal role of genetic factors in determining an individuals susceptibility to this life-threatening condition. The objective of our research was to investigate missense single nucleotide polymorphisms (SNP) associated with MI to determine whether the changes in amino acid sequences have potential implications for the risk of MI. Employing a multifaceted approach, we leveraged an array of computational tools and databases to scrutinize specific missense SNP and meticulously analyzed their potential effects on protein structure stability and function. Our analysis has confirmed a total of 4 missense SNP in ALDH2, APOE, IGFBP1, and PCSK1 genes to be damaging to protein structure and hence, the function. An extensive literature review was then performed to determine the functional roles of these genes in the regulation of the cardiac system-related pathways. Our analysis confirmed that all 2 of these genes are directly involved in pathways related to the cardiac system, while the other 2 genes play other roles. We have further analyzed their interactions and underlying biological processes to determine their potential role in the incidence of MI. These findings collectively offer a profound understanding of the intricate genetic landscape underlying MI. They not only enhance our comprehension of the multifaceted genetic factors influencing MI susceptibility but also set the stage for future experimental investigations. Importantly, these insights hold the potential to guide future research and the development of therapeutic strategies, to improve the prevention and management of this critical cardiovascular condition.

genetics↗

bioRxiv2023

Identification and structural characterization of small-molecule inhibitors of PINK1

Mutations in PTEN-induced putative kinase 1 (PINK1) cause early-onset autosomal recessive Parkinsons Disease (PD). PINK1 is a Ser/Thr protein kinase which functions as a mitochondrial damage sensor and initiates mitochondrial quality control by accumulating on the damaged organelle. There, it will autophosphorylate and then phosphorylate ubiquitin chains, which in turn will recruit and activate Parkin, and E3 ubiquitin ligase also implicated in PD. Ubiquitylation of mitochondrial proteins leads to the autophagic degradation of the damaged organelle. Pharmacological modulation of PINK1 constitutes an appealing avenue to study its physiological function and develop PD therapeutics. In this study, we used a thermal shift assay to identify small-molecule inhibitors of PINK1. In vitro kinase activity assays demonstrate that these molecules are ATP competitive inhibitors that block ubiquitin phosphorylation. PRT062607 (a SYK inhibitor) is the most potent inhibitor of PINK1 in our screen and has an IC50 of 2 M against insect PINK1 and 1 M in HeLa cells expressing human PINK1. The crystal structures of PINK1 in complex with PRT062607 or CYC116 reveal how the compounds interact with the ATP-binding pocket. PRT062607 notably engages with the catalytic aspartate (type-1 inhibition) and causes a destabilization of insert-2 at the autophosphorylation dimer interface. Our findings provide a scaffold for the development of more selective and potent inhibitors of PINK1 that can be used as chemical probes.

biochemistry↗

bioRxiv2023

nELISA: A high-throughput, high-plex platform enables quantitative profiling of the secretome

We present the nELISA, a high-throughput, high-fidelity, and high-plex protein profiling platform. DNA oligonucleotides are used to pre-assemble antibody pairs on spectrally encoded microparticles and perform displacement-mediated detection. Spatial separation between non-cognate antibodies prevents the rise of reagent-driven cross-reactivity, while read-out is performed cost-efficiently and at high-throughput using flow cytometry. nELISA can measure both protein concentration and their post-translational modifications. We assembled an inflammatory panel of 191 targets that were multiplexed without cross-reactivity nor impact on performance vs 1-plex signals, with sensitivities as low as 0.1 pg/mL and measurements spanning 7 orders of magnitude. We then performed a large-scale inflammatory-secretome perturbation screen of peripheral blood mononuclear cells (PBMCs), with cytokines as both perturbagens and read-outs, measuring 7,392 samples and generating [~]1.4M protein data points in under a week; a significant advance in throughput compared to other highly multiplexed immunoassays. We uncovered 447 significant cytokine responses, including multiple putatively novel ones, that were conserved across donors and stimulation conditions. We validate nELISA for phenotypic screening, where its capacity to faithfully report hundreds of proteins make it a powerful tool across multiple stages of drug discovery.

bioengineering↗

bioRxiv2023

Lights, Camera, Mirrors, Action! Toolbox for 3D Reconstruction and Analysis Using a Single Camera and Planar Mirrors

The advent of high-speed cameras being made available for commercial use has facilitated the discovery of intriguing animal behaviors. However, analyzing animal kinematics in three dimensions necessitates using several high-speed cameras, which are specialized equipment and come at a significant expense. In this study, we utilize a conventional methodology of capturing synchronized multiple views using a solitary camera and planar mirrors. We introduce a user-friendly software package that facilitates the calibration of this economical setup and enables the three-dimensional (3D) analysis of high-rate maneuvers. Within[~] 15 minutes, a single high-speed camera is calibrated and can be used to acquire 3D data. We accompany the toolbox with a detailed user guide and video tutorial (https://tinyurl.com/yckm9y6b) for ease of use. We accurately reconstruct the wing beats of multiple flies (dragonfly, butterfly).

animal behavior and cognition↗

bioRxiv2022

Dissecting the interactions of PINK1 with the TOM complex in depolarized mitochondria

Mitochondria dysfunction is involved in the pathomechanism of many illnesses including Parkinsons disease. PINK1, which is mutated in some cases of familiar Parkinsonism, is a key component in the degradation of damaged mitochondria by mitophagy. The accumulation of PINK1 on the mitochondrial outer membrane (MOM) of compromised organelles is crucial for the induction of mitophagy, but the molecular mechanism of this process is still unresolved. Here, we investigate the association of PINK1 with the TOM complex. We demonstrate that PINK1 heavily relies on the import receptor TOM70 for its association with mitochondria and directly interacts with this receptor. The structural protein TOM7 appears to play only a moderate role in PINK1 association with the TOM complex, probably due to its role in stabilizing this complex. PINK1 requires the TOM40 pore lumen for its stable interaction with the TOM complex and apparently remains there during its further association with the MOM. Overall, this study provides new insights on the role of the individual TOM subunits in the association of PINK1 with the MOM of depolarized mitochondria.

biochemistry↗

bioRxiv2021

Selective localization of Mfn2 near PINK1 enable its preferential ubiquitination by Parkin on mitochondria

Mutations in Parkin and PINK1 cause an early-onset familial Parkinsons disease. Parkin is a RING-In-Between-RING (RBR) E3 ligase that transfers ubiquitin from an E2 enzyme to a substrate in two steps: 1) thioester intermediate formation on Parkin, and 2) acyl transfer to a substrate lysine. The process is triggered by PINK1, which phosphorylates ubiquitin on damaged mitochondria, which in turn recruits and activates Parkin. This leads to the ubiquitination of outer mitochondrial membrane proteins and clearance of the organelle. While the targets of Parkin on mitochondria are known, the factors determining substrate selectivity remain unclear. To investigate this, we examined how Parkin catalyzes ubiquitin transfer to substrates. We found that His433 in the RING2 domain catalyzes acyl transfer. In cells, mutation of His433 impairs mitophagy. In vitro ubiquitination assays with isolated mitochondria show that Mfn2 is a kinetically preferred substrate. Using proximity-ligation assays, we show that Mfn2 specifically co-localizes with PINK1 and phospho-ubiquitin in U2OS cells upon mitochondrial depolarization. We propose a model whereby ubiquitination of Mfn2 is efficient by virtue of its localization near PINK1, which leads to the recruitment and activation of Parkin via phospho-ubiquitin at these sites.

biochemistry↗

bioRxiv2021

Structure of PINK1 reveals autophosphorylation dimer and provides insights into binding to the TOM complex

Mutations in PINK1 causes autosomal-recessive Parkinsons disease. Mitochondrial damage results in PINK1 import arrest on the Translocase of the Outer Mitochondrial Membrane (TOM) complex, resulting in the activation of its ubiquitin kinase activity by autophosphorylation and initiation of Parkin-dependent mitochondrial clearance. Herein we report crystal structures of the entire cytosolic domain of insect PINK1. Our structures reveal a dimeric autophosphorylation complex targeting phosphorylation at the invariant Ser205 (human Ser228). The dimer interface requires insert 2, which is unique to PINK1. The structures also reveal how an N-terminal helix binds to the C-terminal extension and provide insights into stabilization of PINK1 on the core TOM complex.

biochemistry↗