Chitosan Defies Death
Gangrene is the death of tissue of certain parts of the body. In Indonesia, people who suffer from Diabetes will also often suffer from Gangrene, which usually affects a patient’s feet. The medication for it is not affordable for everybody. I have chosen the Horseshoe Crab, simply because it’s known widely in Indonesian and can be found easily. Also, due to the fact that, among all crustaceans, the Horseshoe Crab contains the highest levels of Chitin. By using the Chitin found in the shell of the Horseshoe Crab, I shall endeavor to heal the Gangrene of Diabetic Patients.
Antiviral Therapy for Hepatitis C Virus (HCV): Black Mustard Seeds
Hepatitis C Virus (HCV) is an RNA virus, which is considered the main cause of progressive chronic liver disease, cirrhosis, and hepatocellular carcinoma (HCC) worldwide. The number of the patients who are infected with this sleeping virus is increasing rapidly every year, as the unsuitability of the current therapy – interferon α and ribavirin – for most of the genotypes is the main cause of these high rates. Hence, the recent researches are focusing on finding out a new immunotherapy to affect this virus. In this research work, Black Mustard (Brassica nigra) has been used as powdered spice samples to prepare aqueous extracts; One of the included phytochemicals in the black mustard; glucosinolates and their hydrolysis products, was proposed to be used for the HCV patients to prevent the virus progression. Also, the Isothiocyanates are shown with chemotherapeutic and anti-tumor properties. Moreover, some of the structure-related isothiocyanates have the ability to induce the enzyme paraoxonase-1 (PON-1) that is considered hepato-protective agent against liver impairment, inflammation, fibrosis and liver disease mediated by monocyte chemoattractant protein-1 (MCP-1), and is thought to affect the entry of the virus into the hepatocytes. The effect of the black mustard and the produced myrosinase enzyme on the HCV RNA replication is still unknown. In conclusion, the black mustard is thought to affect the progression and the fluidity of the HCV envelope resulting in impairment of viral binding and fusion.
Novel Approach to Screening Mutations Causing Retinoblastoma, a Childhood Cancer of Retina
Retinoblastoma (RB) is a childhood retinal cancer caused by mutations in the RB1 gene. Molecular diagnosis is crucial for early detection and treatment. Current DNA diagnostic screening requires substantial amounts of tumour and blood samples. However current screening methods face the challenges of limited DNA templates from minute retinal tumours and too much blood samples drawn from young patients. In addition, the starting DNA template amount and quality are important to ensure confident detection of disease-causing mutations. As the majority of RB1 mutations are unique and distributed throughout the RB1 gene with no real hot spots, the entire gene needs to be thoroughly analysed. This investigation proposes to enrich DNA samples using a whole genome amplification (WGA) step prior to RB1 mutation screening by RB1 gene-specific PCR amplification as well as high resolution melt (HRM) analysis and sequencing. It also identifies RB1 mutations in two RB patients and explores whether WGA and saliva products can be a source of DNA templates for RB1 analysis. In addition, this study was conducted based on the hypotheses that RB1 mutations were the underlying cause of the disease in the two patients, and that the products from WGA could be used specifically for RB1 gene analysis to overcome the constraint of insufficient DNA samples. Two anonymised genomic DNA samples from two unrelated RB patients and five normal healthy DNA samples were used in this project. WGA kits were compared according to three criteria, namely amplification yield, product fragment size and whether DNA is amplifiable. Prior to and after amplification, the optical density of two normal samples was measured to determine the increase in DNA yield. The amplicons were subjected to gel electrophoresis to determine the product fragment size. Exons 6, 14 and 25 of the original and amplified samples undergone PCR, and were examined again using gel electrophoresis to ascertain that the amplicons were amplifiable. Mutation analysis using HRM was carried out with pre-existing primers for all 27 exons and the promoter of RB1. Samples from patients were analysed against 83 saliva DNAs extracted using Oragene•DNA (OG-500) Kit. REPLI-g was observed to produce higher yield and products of reliable fragment size. Single distinct bands were also seen for exons amplified using REPLI-g, indicating that REPLI-g is more accurate and suitable in the amplification of DNA. Abnormal melt profiles were obtained for exon 6 in RB477 and exon 14 in RB572 for HRM. These exons were sequenced to determine the exact mutation. Exon 6 was found to have a splice-site mutation g.607+1G>T, while a point mutation, g.1363C>T (p.Arg455X) was identified in exon 14. Both the uses of saliva as a non-invasive DNA source and the WGA approach for enriching DNA sample for application in RB1 gene analysis have never been reported for RB. Although HRM analysis has been used for other diseases, this is its first instance applied in work on RB1 gene. In short, this report offers novel and promising approaches which would contribute significantly to the molecular analysis of mutations in RB.
BA-ADA based ROS-responsive nanoparticles for selective drug delivery in cancer cells
Current medical intervention in cancer therapeutic methods has shown risks and side effects with normal tissues. This includes incomplete cancer eradication. In reference to numerous studies and literature reviews, a stimuli-responsive drug delivery system is selected as an innovative, safe and more assured treatment due to its site-specific release ability. This allows specific intervention upon the given stimulus which response to the presenting disease symptoms. Hence, we designed a ROS(Reactive Oxygen Species)-responsive BA-ADA(4-Hydroxyphenylboronic acid pinacol ester and 1-Adamantanecarboxylic acid bonded molecule) nanoparticle delivery system. In our study, ROS-responsive nanoparticle was designed and prepared based on a synthetic molecule from BA and ADA. A therapeutic payload, Doxorubicin, can be loaded into the nanoparticles and it can be selectively released within cancerous tissues whereby ROS level is over-expressed. This will enhance both therapeutic efficiency and reduce side effects. The stability and ROS-responsiveness of the particle were proven in a series of evidence-based experiments. The results showed a significant difference in cell viability during the experiments with healthy and cancerous cell samples. Further research will be required to extend the experiment in vivo.