THE DESIGN OF MICROFLUIDIC PUMP (MFP) FOR MEDICAL FIELD
The ability of microfluidic (MF) device technologies to provide a lot of information with a small amount of sample, the opportunities it offers increases their use in the medical field in the bedside monitoring in drug delivery systems. Three-dimensional (3D) printer technologies provide advantages such as cost-effectiveness in the production of MF devices and quick and easy production in intricate designs. In our project, it is aimed to design microfluidic pumps (MFP) to be used in the medical field and conduct its production with 3D printer technologies. The developed MFP is intended to be at low cost, bio-compatible, adaptable, and portable to the drug, suitable flow properties as a pharmaceutical pump. First of all, MFP air channel, flow channel, etc. parts were designed and printed with the help of a 3D printer and on AutoCAD, one of the professional drawing programs. The poly(dimethylsiloxane) (PDMS) membrane that will enable MFP activation is produced in different thicknesses and glued to the air channel of MFP. The resistance to the applied pressure is observed, and the appropriate membrane thickness is determined as ~ 235µm. Liquid PDMS was applied to the inner surfaces of MFP's air and flow channel, PDMS membrane was placed between them, and the parts were assembled in the oven at 60ºC. MFP has been connected to the pneumatic valve system, where operation codes have been prepared with Arduino Uno, and flow properties have been examined. The flow rate of MFP is ~ 50 µL/min at a maximum of 15 Hz, and the backpressure is ~ 0.085 Pa under a maximum pressure of 3 bar. Also, values such as size, membrane thickness, and applied pressure for the possible models of MFP were supported by theoretical calculations. As a result, MFP, which is biocompatible, drug adaptable, portable, wearable technology application potential, and has suitable flow characteristics as a pharmaceutical pump, has been developed. MFP introduced a microfluidic pump system that can make life easier for the patient and contribute to the national economy through domestic production and can be used as a drug pump in the treatment of diseases such as diabetes and cancer.
HOST TARGET PROTEINS OF SPIKE PROTEIN OF SARS-COV-2
Coronavirus Disease 2019 (COVID-19) is a newly emerged infectious disease caused by the new severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV-2). In less than one year, the virus has spread around the entire world, killing millions of people and disrupting travel and business worldwide. During infection, the virus uses its Spike protein to dock onto the Ace2 protein on the surface of its human host cell. Spike is 1273 amino acids long and only a short fragment of Spike (319-541) is sufficient to bind Ace2. We hypothesized that the remaining protein sequences of Spike might have functions for viral replication beyond the binding of Ace2. We have performed Split-Ubiquitin protein-protein interaction screens to isolate human proteins by their ability to bind to Spike, and we have identified Annexin2A2 and Cytochrome b as novel human protein interaction partners of Spike. Annexin2A2 is involved in both endocytosis and exocytosis, and the protein interaction with Spike might help the virus to enter and exit its host cell. The presence of the mitochondrial Cytochrome b protein inside the cytosol promotes apoptosis, and the protein interaction with Spike could speed up sapoptosis of the infected human cell. The Nub cDNA libraries that we have generated also allowed us to screen for synthetic peptides that interact with Spike. We have isolated two synthetic peptides, FL1a and FL7a, derived from the non-coding parts of human mRNAs by their ability to interact with Spike. We found that both FL1a and FL7a interact with the C-terminal half of the Spike protein. We also found that FL7a is able to block the Spike-Spike self-interaction at the C-terminal half of the Spike protein and we think that this could block the reassembly of the Spike protein in the host cell during viral reassembly. We hope that those synthetic peptides could be used as drugs due to their ability to block protein-protein interactions of Spike with human host proteins that are essential for viral replication.
Preparation of a Specific Detector for Aspergillus Niger in Swimming Pools
Swimming pools are one of the transmission routes of superficial and cutaneous fungal infections. Maintenance of environmental hygiene in different parts of swimming pools is of great importance, especially the hygiene of water (1). The conventional fungal detection methods include direct smear preparation, culture, and pathological examinations. However, these methods are not fast enough or do not have sufficient sensitivity (2). Therefore, the present research introduces a novel method for detecting Aspergillus niger in pool water through creating optimal conditions for this fungus, which leads to the citric acid production by the fungus and pH changes of the related culture medium. Four experiments in 10 steps were performed to find the optimal conditions for fungal growth. According to our results, adding each of the variables sucrose, soy, and ferrous sulfate can lead to favorable results. Moreover, the shaker speed increase and fungal aeration are important. Also, we showed that soybean led to the best results compared to other variables. Considering the obtained results, including the shortened detection duration and cost-effectiveness, this method can be presented to the swimming pool owners and pathobiology laboratories as the method of choice for Aspergillus niger detection.
HOST TARGET PROTEINS OF SPIKE PROTEIN OF SARS-COV-2
Coronavirus Disease 2019 (COVID-19) is a newly emerged infectious disease caused by the new severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV-2). In less than one year, the virus has spread around the entire world, killing millions of people and disrupting travel and business worldwide. During infection, the virus uses its Spike protein to dock onto the Ace2 protein on the surface of its human host cell. Spike is 1273 amino acids long and only a short fragment of Spike (319-541) is sufficient to bind Ace2. We hypothesized that the remaining protein sequences of Spike might have functions for viral replication beyond the binding of Ace2. We have performed Split-Ubiquitin protein-protein interaction screens to isolate human proteins by their ability to bind to Spike, and we have identified Annexin2A2 and Cytochrome b as novel human protein interaction partners of Spike. Annexin2A2 is involved in both endocytosis and exocytosis, and the protein interaction with Spike might help the virus to enter and exit its host cell. The presence of the mitochondrial Cytochrome b protein inside the cytosol promotes apoptosis, and the protein interaction with Spike could speed up sapoptosis of the infected human cell. The Nub cDNA libraries that we have generated also allowed us to screen for synthetic peptides that interact with Spike. We have isolated two synthetic peptides, FL1a and FL7a, derived from the non-coding parts of human mRNAs by their ability to interact with Spike. We found that both FL1a and FL7a interact with the C-terminal half of the Spike protein. We also found that FL7a is able to block the Spike-Spike self-interaction at the C-terminal half of the Spike protein and we think that this could block the reassembly of the Spike protein in the host cell during viral reassembly. We hope that those synthetic peptides could be used as drugs due to their ability to block protein-protein interactions of Spike with human host proteins that are essential for viral replication.
Anti-bacterial Crab bio-bandages with Bio-dressings 2.0
Commercially available bandages such as hydrocolloid are neither biodegradable nor anti-bacterial. Chitin is known to be the second most naturally available polysaccharide which could be transformed to chitosan which is known to be anti-bacterial (Hasan, 2018) (Chao, 2019) and haemostatic (Okamoto, 2003) (Hu, 2018). Chitosan can be further converted to hydrogel which is bio-degradable and has good water absorbance. Anti-bacterial crab bio-bandages and crab bio-dressings should be bio-degradable as it took 42 days and a month for complete bio-degradation respectively, so they should be better than commercial bandages such as Nexcare Hydrocolloid as the disposal of anti-bacterial crab bio-bandages with bio-dressings would no longer pose burden to landfilling or threat to our environment. Anti-bacterial crab bio-bandages with bio-dressings are anti-bacterial with degree of deacetylation of DD% (measured using FTIR Spectrum II) 82.6% (due to the presence of chitosan) even without the application of other anti-bacterial agents and hence can provide complete protection of wounds from skin and soft tissues infections and haemostatic (due to the presence of chitosan). After testing and certification based on IS997:2004 and BS EN 13726-1, they should meet many requirements specified. Anti-bacterial crab bio-bandages should be eligible for marketing. Some results were as follows: 1.4 Anti-bacterial effect of crab hydrogels and roasted crab hydrogels Pure chitosan, crab chitosan, crab hydrogels and roasted crab hydrogels showed significant anti-bacterial effect. NO oral bacterial colonies were present in drinking water with crab hydrogels. Thus crab hydrogels could serve as effective anti-bacterial wound dressings. 1.6 Basing on IS997:2004 standard, the load per unit of area of anti-bacterial bio-bandages was 342g/m2 which met the minimum requirement of 36g/m2, the anti-bacterial bio-bandages had stronger tension strength (>20N both in dry and wet conditions) than commercial hydrocolloid. (2.7N dry 2.8N wet) which was comparable with that required (50-67N) and pH of about 7 which met the pH range of 4.5-8. 1.7 The FSA Free-Swell Absorbency of synthetic blood of crab hydrogel bio-dressings was 1.86g per 5cm x 5cm dressing which was much higher than that of commercial hydrocolloid (0.299g per 5cm x 5cm dressing) based on BS EN 13726-1.