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

Environmental radiation as the conditioning factor for the survival of yeast <i><span style="font-size:14.0pt;line-height:115%;font-family:"Times New Roman"; mso-fareast-font-family:"Times New Roman";color:black;mso-ansi-language:EN-IN; mso-fareast-language:EN-IN;mso-bidi-language:HI" lang="EN-IN">Saccharomyces cerevisiae</span></i>

95-100Whether natural radiation can be a conditioning factor for the growth and survival of a living organism was investigated using diploid yeast  S. cerevisiae D7. Yeast cells were conditioned by growing them continuously for at least 100 generation in 3 different radiation background such as i) ambient radiation (1.1 mSv/y), ii) sub-ambient radiation (0.44 mSv/y, within a shielded chamber) and iii) an elevated background radiation (88 and 880 mSv/y in a γ-field). At the end, the cells were challenged with 60Co γ, 100 Gy and the viable fractions were determined. Conditioning the cells in 880 mSv/y and in ambient radiation, enabled the cells to reduce the deleterious effect of the challenging dose significantly (P < 0. 05) compared to that of sub-ambient radiation. The cellular viability of yeast cultures seems to be influenced by the prevailing radiation background, apart from starvation. Comparatively, a rapid decline in viability was noticed when the cultures were incubated for 60 days in the shielded chamber. The results indicate that some amount of radiation equivalent to background level or little above is needed to confer fitness in biological systems against stress factors, including radiation. The adaptive dose for the diploid yeast was also determined by single exposure. The priming dose ranged from 0.01 to 1.2 Gy. An adaptive dose of 0.25 or 0.4 Gy, almost nullified the deleterious effect of the challenging dose. The adaptive response may have a greater role in the field of cancer therapy and in radiation risk assessment. Understanding the response of an organism at different radiation-background will be helpful for successful space management<span style="font-size:14.0pt;line-height: 115%;font-family:Fd8138-Identity-H;mso-fareast-font-family:" times="" new="" roman";="" mso-bidi-font-family:fd8138-identity-h;color:black;mso-ansi-language:en-in;="" mso-fareast-language:en-in;mso-bidi-language:hi"="" lang="EN-IN">.</span

Standardisation and evaluation of DNA-lanthanide fluorescence spectroscopy for determining rifampicin resistance in Mycobacterium tuberculosis clinical isolates

SETTING: Tuberculosis Research Centre, Chennai. OBJECTIVE: To rapidly identify multidrug-resistant Mycobacterium tuberculosis using a novel method. DESIGN: A new assay, based on DNA-lanthanide fluorescence, was standardised and evaluated using 93 each of coded rifampicin-resistant and rifampicin-sensitive M. tuberculosis clinical isolates for the correct identification of rifampicin resistance. The results obtained by the new assay were compared with the conventional results. RESULTS AND CONCLUSION: The new assay gave a sensitivity and specificity of 88% and 85%, respectively. It is simple, easy to perform and requires 48 hours for the drug susceptibility results to be available after obtaining the primary culture

Activation of a lateral hypothalamic-ventral tegmental circuit gates motivation.

Across species, motivated states such as food-seeking and consumption are essential for survival. The lateral hypothalamus (LH) is known to play a fundamental role in regulating feeding and reward-related behaviors. However, the contributions of neuronal subpopulations in the LH have not been thoroughly identified. Here we examine how lateral hypothalamic leptin receptor-expressing (LHLEPR) neurons, a subset of GABAergic cells, regulate motivation in mice. We find that LHLEPR neuronal activation significantly increases progressive ratio (PR) performance, while inhibition decreases responding. Moreover, we mapped LHLEPR axonal projections and demonstrated that they target the ventral tegmental area (VTA), form functional inhibitory synapses with non-dopaminergic VTA neurons, and their activation promotes motivation for food. Finally, we find that LHLEPR neurons also regulate motivation to obtain water, suggesting that they may play a generalized role in motivation. Together, these results identify LHLEPR neurons as modulators within a hypothalamic-ventral tegmental circuit that gates motivation

"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