Recently, a protocol for ghost cells preparations was introduced. It was given the name sponge-like protocol: Procaryotes, eucaryotes and virus were turned to ghost cells using such protocol. In this study, with slight modifications, Aspergillus niger ghost cells were prepared using the same protocol. Both the Minimum Inhibitory Concentration (MIC) and the minimum growth concentration (MGC) values for H 2 O 2 , NaOH, NaHCO 3 and SDS against A. niger were determined. Five different randomization experiments were conducted instead of the full Plackett-Burman design. During the ghost preparation steps, the released Protein and DNA were measured spectrophotometrically at 280nm and 260nm, respectively. The quality of the prepared ghost cells were evaluated during the preparation steps using light microscope. Transmission electron microscope was used for evaluating the final steps. Protein and DNA electrophoresis were conducted to evaluate the quality of the released protein and DNA after each randomization experiment. The data obtained prove correct evacuation of the fungal cells from their cytoplasmic content during the successive steps. The study not only introduces a protocol for preparing ghost cells from Aspergillus niger but also enables the isolation of both of protein and DNA. The idea, the concept and the tools used in this study could establish a more sensitive method for protein and DNA isolation using any of four utilized chemical compounds. This proposes the same concept of enzyme-induced cell lysis which is based on minimizing the effect of used chemicals or enzymes. The study recommended extending the benefit of the sponge-like protocol from being a protocol for ghost cells preparation to DNA and protein isolation technique using the same concept.
There is an increasing interest toward the understanding of the important fungus, Aspergillus flavus, which has a high impact on our health and economy. Controlling such microbe could be achieved by evacuating its cells using simple tool. Recently, the Sponge-Like protocol for bacterial ghosts' preparation was introduced. Then, this protocol concept was extended to include not only bacterial cells but also yeast cells as well as viruses. In this study, for the first time we applied this protocol to pathogenic fungi as Aspergillus flavus turning it to unviable, dead and evacuated ghost cells. We introduce the idea of using the minimum growth concentration (MGC) and the minimum inhibition concentration (MIC) for controlling Aspergillus flavus and other type of fungi either in our food or those infecting us. Using mixture of compounds proves to have effect on certain microbes upon the use of their minimum effect, which could be a solution for treating many diseases without damaging the cells and tissues as well as our organs. A. flavus which is responsible for causing different superficial infections besides its ability to produce hazardous aflatoxins was used as a model in this study. The future will show an increasing interest in this protocol either for ghost cells preparation or controlling pathogens with minimum side effects of the used destabilizing and killing compounds or drugs. Any chemical compound that could achieve the same goal is invited. We strongly recommend and encourage conducting research in this field using the same or similar tactics and idea.
A massive number of Literature is accessible focusing only on the general concern of climate change ignoring the cost valuation of the association between environmental change and human health. Establishment of environment-disease relationship leading to economic consequences is found timely and apposite research at this moment. Therefore, this paper aims to establish environmental-health relationship and its impact on household economy through cost of illness in western part of Nepal. Time series analytical economic evaluation design is employed using secondary data to establish environment-disease relationship with the help of ordinary least square, being based on the econometric theory. The results divulge that environmental variables such as extreme summer temperature warm winter temperature and population density are in favor to increase the disease prevalence, whereas, increasing winter rainfall, immunization coverage and vitamin A coverage among children seemed to reduce the disease prevalence in western Nepal. Similarly, disease-environment relationship showed that 1 0 C increase in temperature increases the disease prevalence at least by 5% in the existing rate of prevalence in western Nepal, controlling other variables.Above relation in sensitivity analysis assuming linear relationship showed that every 0.5 0 C increase in temperature leads to increase in cost of illness at household level at least by NPR 77.7 million in western Nepal. Establishment of environment-disease relationship concludes that adaptation measures for protecting from summer extreme temperature is an urgent action needed for disease reduction in western Nepal. Besides, controlling population, extension of immunization and vitamin A programs to the children are also seemed to reduce illness in western Nepal. Therefore, recommendations are made accordingly.
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