Expression and purification of Murine IFN-γ protein from cloned E. coli strain containing pRSET A Vector with IFN gamma gene

The cloned E. coli cell containing Murine IFN -γ inserted pRSET A vector system was effectively expressed in this study.  The induction of the clones was done using IPTG in E.coli and induces mRNA generation and synthesis protein. It has shown an expression of protein with 18 kda in SDS PAGE and western blotting and their size was determined by GENE RUNNER software.  This recombinant protein has a 6x His tag and it has been proved as it has shown a potent anti His property in western blotting. The purification of the protein was further done by Ni-NTA affinity chromatography. Nitrilo tri acetic acid (NTA) binds more stably with nickel (Ni) with 4 to 6 ligand binding sites in the coordination sphere of Nickel leaving two sites free to interact with the 6X His tag. The total results conclude that the targeted IFN gamma (408bp mouse gene) cloned in pRSET A was effectively expressed in E. coli BL21 strain cells and purified IFN gamma protein effectively as 1mg/ml. The purified IFN gamma protein may be used to diagnose the antiviral activity and antitumor activity. Key words: IFN gamma, pRSET A, E. coli, SDS PAGE, Western Blottin

Parkin-deficient Mice Exhibit Nigrostriatal Deficits but not Loss of Dopaminergic Neurons

Loss-of-function mutations in parkin are the major cause of early-onset familial Parkinson's disease. To investigate the pathogenic mechanism by which loss of parkin function causes Parkinson's disease, we generated a mouse model bearing a germline disruption in parkin. Parkin-/- mice are viable and exhibit grossly normal brain morphology. Quantitative in vivo microdialysis revealed an increase in extracellular dopamine concentration in the striatum of parkin-/- mice. Intracellular recordings of medium-sized striatal spiny neurons showed that greater currents are required to induce synaptic responses, suggesting a reduction in synaptic excitability in the absence of parkin. Furthermore, parkin-/- mice exhibit deficits in behavioral paradigms sensitive to dysfunction of the nigrostriatal pathway. The number of dopaminergic neurons in the substantia nigra of parkin-/- mice, however, is normal up to the age of 24 months, in contrast to the substantial loss of nigral neurons characteristic of Parkinson's disease. Steady-state levels of CDCrel-1, synphilin-1, and α-synuclein, which were identified previously as substrates of the E3 ubiquitin ligase activity of parkin, are unaltered in parkin-/- brains. Together these findings provide the first evidence for a novel role of parkin in dopamine regulation and nigrostriatal function, and a non-essential role of parkin in the survival of nigral neurons in mice

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Green synthesis of multifunctional PEG-carboxylate π back-bonded gold nanoconjugates for breast cancer treatment

Mani Gajendiran,1,2 Heejung Jo,2 Kyobum Kim,2 Sengottuvelan Balasubramanian1 1Department of Inorganic Chemistry, University of Madras, Guindy Campus, Chennai 600025, India; 2Division of Bioengineering, School of Life Sciences and Bioengineering, Incheon National University, Incheon 22012, Republic of Korea Background: Surface functionalization of gold nanoparticles (AuNPs) has emerged as a promising field of research with enormous biomedical applications. The folate (FA)-attached polymer-gold nanoconjugates play vital role in targeting the cancer cells.Methods: AuNPs were synthesized by using di- or tri-carboxylate-polyethylene glycol (PEG) polymers, including citrate-PEG (CPEG), malate-PEG (MAP), and tartrate-PEG (TAP), as a reducing and stabilizing agent. After synthesis of polymer-AuNPs, the freely available hydroxyl and carboxylate groups of CPEG, MAP, and TAP were used to attach a cancer cell-targeting agent, FA, via a 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxy succinimide coupling reaction to obtain FA-CPEG-AuNP, FA-MAP-AuNP, and FA-TAP-AuNP nanoconjugates, respectively. The 5-fluorouracil (5FU) was attached to π back-bonded carbonyl oxygens of the nanoconjugates, and the in vitro drug release profile was studied by high pressure liquid chromatography. Biocompatibility profiles of the FA-CPEG-AuNP, FA-MAP-AuNP, and FA-TAP-AuNP nanoconjugates were investigated using adult human dermal fibroblasts. Anti-breast cancer activity of 5FU-loaded nanoconjugates was investigated using MCF-7 breast cancer cells.Results: X-ray photoelectron spectroscopy and Fourier-transform infrared spectroscopy analyses confirmed that AuNPs attached to CPEG, MAP, or TAP via the formation of π back bonding between AuNPs and the ester carbonyl group. The π back-bonded nanoconjugates exhibited sustained release of 5FU up to 27 days. FA-MAP-AuNPs exhibited an IC50 at 5 µg/mL, while FA-CPEG-AuNPs and FA-TAP-AuNPs showed the IC50 at 100 µg/mL toward MCF-7 cancer cells.Conclusion: The developed polymer π back-bonded multifunctional gold nanoconjugates could be used as a potential drug delivery system for targeting MCF-7 cancer cells. Keywords: polymer-gold nanoconjugates, 5-fluorouracil, anticancer activity, MCF-7 cells, green synthesi

"Adaptive response" - some underlying mechanisms and open questions

Organisms are affected by different DNA damaging agents naturally present in the environment or released as a result of human activity. Many defense mechanisms have evolved in organisms to minimize genotoxic damage. One of them is induced radioresistance or adaptive response. The adaptive response could be considered as a nonspecific phenomenon in which exposure to minimal stress could result in increased resistance to higher levels of the same or to other types of stress some hours later. A better understanding of the molecular mechanism underlying the adaptive response may lead to an improvement of cancer treatment, risk assessment and risk management strategies, radiation protection, e. g. of astronauts during long-term space flights. In this mini-review we discuss some open questions and the probable underlying mechanisms involved in adaptive response: the transcription of many genes and the activation of numerous signaling pathways that trigger cell defenses - DNA repair systems, induction of proteins synthesis, enhanced detoxification of free radicals and antioxidant production.Publisher PDFPeer reviewe

Impact of calcined temperatures on the crystalline parameters, morphological, energy band gap, electrochemical, antimicrobial, antioxidant, and hemolysis behavior of nanocrystalline tin oxide

To construct a battery, the precipitation-synthesized SnO2 products at 450 °C and 650 °C were separately taken and mixed with graphite as the anode and PbO2,V2O5, and graphite materials as cathode materials to make the pellets and examine their open circuit voltage (OCV) values. The microstrain, lattice parameter, and crystallite size values of the above-mentioned tin oxide compounds were obtained through Rietveld refinement-MAUD fit analysis. The microstrain and lattice parameter values of tin oxide were significantly varied at a higher calcined temperature. Surface particle grain growth was increased with the increased calcined temperature from 450 to 650°C as evidenced by FE-SEM study. Particle size distributions of SnO2 and polycrystalline behavior have been discussed with the aid of TEM analysis. From the UV-visible spectra, optical band gap (Eg) values reduced from 3.73 to 3.69 eV for the SnO2 products with an increase in calcined temperatures from 450 to 650 °C. The antimicrobial responses of the two different calcined SnO2 samples at 450 °C and 650 °C against two different bacterial pathogens (gram-positive-S. aureus and gram-negative-E.coli) were investigated. From the microbicidal assessment, a relatively higher diameter of the zone of inhibition (DZOI) of tin oxide at 650°C samples was measured to be 19 ± 2 mm and 21 ± 2 mm for S. aureus and E. Coli than the DZOI of SnO2 at 450 °C samples (15 ± 1 mm for S. aureus and 18 ± 1 mm for E. coli. DPPH scavenging activity at 100 μg/ml shows that SnO₂ calcined at 450 °C achieves 68 ± 1%, while SnO2 calcined at 650 °C exhibits a significantly higher activity of 86 ± 1%. A slight increase in hemolysis was observed for SnO2 calcined at 650°C, reaching 1.3% at higher concentrations, but overall, hemolysis remained below 5%, indicating high hemocompatibility