In the last several decades, the number of people dying from cancer-related deaths has not reduced significantly despite phenomenal advances in the technologies related to diagnosis and therapeutic modalities. The principal cause behind limitations in the curability of this disease is the reducing sensitivity of the cancer cells towards conventional anticancer therapeutic modalities, particularly in advance stages of the disease. Amongst several reasons, certain secretory factors released by the tumour cells into the microenvironment have been found to confer resistance towards chemo- and radiotherapy, besides promoting growth. Interleukin-6 (IL-6), one of the major cytokines in the tumour microenvironment, is an important factor which is found at high concentrations and known to be deregulated in cancer. Its overexpression has been reported in almost all types of tumours. The strong association between inflammation and cancer is reflected by the high IL-6 levels in the tumour microenvironment, where it promotes tumorigenesis by regulating all hallmarks of cancer and multiple signalling pathways, including apoptosis, survival, proliferation, angiogenesis, invasiveness and metastasis, and, most importantly, the metabolism. Moreover, IL-6 protects the cancer cells from therapy-induced DNA damage, oxidative stress and apoptosis by facilitating the repair and induction of countersignalling (antioxidant and anti-apoptotic/pro-survival) pathways. Therefore, blocking IL-6 or inhibiting its associated signalling independently or in combination with conventional anticancer therapies could be a potential therapeutic strategy for the treatment of cancers with IL-6-dominated signalling.
Metabolic viability based high throughput assays like MTT and MTS are widely used in assessing the cell viability. However, alteration in both mitochondrial content and metabolism can influence the metabolic viability of cells and radiation is a potential mitochondrial biogenesis inducer. Therefore, we tested if MTT assay is a true measure of radiation induced cell death in widely used cell lines. Radiation induced cellular growth inhibition was performed by enumerating cell numbers and metabolic viability using MTT assay at 24 and 48 hours (hrs) after exposure. The extent of radiation induced reduction in cell number was found to be larger than the decrease in MTT reduction in all the cell lines tested. We demonstrated that radiation induces PGC-1α and TFAM to stimulate mitochondrial biogenesis leading to increased levels of SDH-A and enhanced metabolic viability. Radiation induced disturbance in calcium (Ca2+) homeostasis also plays a crucial role by making the mitochondria hyperactive. These findings suggest that radiation induces mitochondrial biogenesis and hyperactivation leading to increased metabolic viability and MTT reduction. Therefore, conclusions drawn on radiation induced growth inhibition based on metabolic viability assays are likely to be erroneous as it may not correlate with growth inhibition and/or loss of clonogenic survival.
Clay minerals such as kaolinite, smectite, chlorite, micas are main components of raw materials of clay and formed in presence of water. A large number of clays used to form the different structure which completely depends on their mining source. They are known as hydrous phyllosilicate having silica, alumina and water with variable amount of inorganic ions like Mg2+, Na+, Ca2+ which are found either in interlayer space or on the planetary surface. Clay minerals are described by presence of two-dimensional sheets, tetrahedral (SiO4) and octahedral (Al2O3). There are different clay minerals which are categorized based on presence of tetrahedral and octahedral layer in their structure like kaolinite (1:1 of tetrahedral and octahedral layers), smectite group of clay minerals (2:1 of tetrahedral and octahedral layers) and chlorite (2:1:1 of tetrahedral, octahedral and octahedral layers). The particle size of clay minerals is <2microns which can be present in form of plastic in presence of water and solidified when dried. The small size and their distinctive crystal structure make clay minerals very special with their unique properties including high cation exchange capacity, swelling behavior, specific surface area, adsorption capacity, etc. which are described in this chapter. Due to all these unique properties, clay minerals are gaining interest in different fields.
RAPD is a PCR based technique which involves the use of single arbitrary short primers (8-12 nucleotides), resulting in the amplification of many discrete DNA. The segments of DNA that are amplified are random. The technique was developed independently by two different laboratories and called as RAPD and AP-PCR (Arbitrary Primed PCR). This procedure detects nucleotide sequence polymorphisms in a DNA amplification based assay using only a single primer of arbitrary nucleotide sequence. The RAPD technology has provided a quick and efficient screen for DNA-sequence polymorphisms at a very large no of loci. The present communication gives emphasis on basic knowledge about RAPD, procedure, its advantages disadvantages, limitations and applications of RAPD.
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