2.1. Fish Scales Preparation and Demineralization:
Fish Scales of Nile Tilapia, caught from Nile Delta, were randomly collected from the local market during the month of June. The scales were washed thoroughly with distilled water, and stored at –20°C until used. All reagents used were of analytical grade. Fish scales were washed twice in 10 wt.% of NaCl solutions (scales: solution = 1:10) to remove non-collagenous proteins on the surface by stirring the solution for 24h, then washed thoroughly with distilled water until the pH was neutral. Demineralization was achieved by stirring scales for 90 min. in 0.4 mol/l HCl (scales: solution = 1:15), followed by washing the scales for three times in distilled water to be ready for collagen extraction.
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The collagen samples were dissolved in 0.1 M acetic acid with different dilutions. The dissolved collagen samples were mixed with the sample buffer (0.5 M Tris-HCl buffer pH 6.8 which contained 30% (v/v) glycerol, 5% (w/v) SDS, 10% (v/v) β-mercaptoethanol, and 0.04% (w/v) Bromophenol blue). Coo- massie Brilliant Blue G-250 was used to visualize the gel after the electrophoresis. High molecular weight protein markers were used to estimate the molecular weight of proteins.
2.3.2 Amino Acid Analysis:
The amino acid composition of collagen was analyzed with high performance amino acid analyzer (Biochrom 30, Biochrom Limited, Cambridge, England). Lyophilized samples were dissolved in 6 M HCl solutions and hydrolyzed at 110°C for 24 h.
Hydroxyproline content was detected using The GC/MS QqQ system (Agilent 7890A series GC system coupled with an Agilent 7000B QqQ MS, Agilent Technologies UK Ltd). Derivatization has been done according to the protocol developed by Villas-Boas et al. [23]. A Standard spectrum was first done using hydroxyproline standard, to determine the mass spectrum of hydroxyproline, which was later used to quantify hydroxyproline.
2.3.3 Determination of Denaturation Temperature
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BHK-21 cells were seeded on the 24-well collagen gel coated plate, containing Hank 's Balanced Salt Solution (HBSS) with Phenol Red (BioWhittaker™, Lonza, Belgium), supplemented with 10% fetal bovine serum (FBS) (Gibco Invitrogen, Milan, Italy), with 1% penicillin/ streptomycin (Sigma-Aldrich, Inc., St. Louis, USA), at density of 3×104 cells per well. Some wells were kept untreated as controls to compare with tests. The medium was changed every 2 days. At days 1 and 2, the cells on the collagen gels were observed by an inverted light microscope (DMI3000 B; Leica,
Crayfish are decapods pertaining to the phylum arthropoda, which are invertebrates and contain an open-circulatory system.1 The system works by the hemolymph getting re-oxygenated in the gills before being transporting to the heart by brachio-cardiac veins and then pumped to sinuses that bathe tissues with oxygenated hemolymph.2 The crayfish contain a neurogenic heart that depends on neuronal input from cardiac ganglion.3 The neurogenic heart requires nerve impulses to produce contractions in contrast to a myogenic heart that can contract independently from the nervous system.3 The crayfish heart will beat due to the reaction of the ganglion to stimuli in the environment.4 Stimuli will affect the autonomic nervous system in the crayfish that control involuntary actions such as the heart rhythm.4 Neurotransmitters are the chemical signals in which the nervous system regulates both heart rate and contraction.3The autonomic nervous system breaks into two categories as parasympathetic system that is involved in relaxation of organs and the sympathetic system that will stimulate increased activity. Neurotransmitters can either increase or decrease heart rate by altering the patterns in neural activity of the heart.4 Crayfish are poikilotherms, which means they cannot metabolically thermoregulate thus conform to the ambient temperature in the water.
The temperature probe was kept in the calorimeter until the temperature had been stabilized and was calibrated. A beaker was placed on a hot plate with dial turned between three and four. Another 100.00 ml of deionized water was added while the beaker is heating up. Using the temperature probe, the beaker was measured
Blackfish, a documentary by Gabriela Cowperthwaite, a historically observed impact of the captivity of killer whales. This Documentary focuses on the orca Tilikum, who was captured off the coast of Iceland. On February 21, 1991, Sealand trainer Keltie Byrne fell into the pool with Tilikum. She was pulled to the bottom by Tilikum, tossed around, and drowned. It took Sealand employees two hours to recover her.
Unknown Lab Report Unknown # 25 By: Jenna Riordan March 19, 2018 Bio 2843 1. Introduction Microbiology is the study of microorganisms found in all different environments throughout Earth, from the hot thermal vents at the bottom of the ocean to the ice at the top of a mountain.
Imagine surviving a fatal experience only to be punished and blamed for a death you could do nothing about. Now, imagine losing a girlfriend and almost losing a sister and having to live with the grief with no one to share your pain. These are the two situations Sage and Maddy find themselves in after Isabel’s death (Sage’s girlfriend) in “The Isabel Fish” by Julie Orringer. Isabel died in a car crash with Maddy in the passenger seat, and she has felt blamed by everyone, including her brother and Isabel’s friends. Because of Isabel’s death, Maddy 's relationship with her brother became a series of punishments, however, after Sage feels guilty for killing Maddy 's fish, they finally opened up to each other and their relationship strengthens.
What is life for an orca really like at SeaWorld? It’s sad, degrading, confining, and tortuous with perhaps a few small glimpses of happiness, depending on the whale and the day. According to the documentary Blackfish, Tilikum, or Tilly as some may have known him as, was an orca whale captured and kept in captivity at SeaWorld, until his death in early January 2017. The documentary reveals that Tilly was taken from his mother as a mere infant. People used nets to corral mother orcas and their young so that the babies would be the only ones to be caught.
Firstly, it was readily available one of the few pure proteins
How can anyone look at the whale crying and think that is morality because it is not. Whale ate one of the trainers. Tilikum is responsible for the deaths of 3 individuals, including a top killer whale trainer, Dawn Brancheau. Blackfish shows the devastating consequences of keeping such intelligent and sentient creatures in captivity. They knew Tilikum was dangerous because in his profile information it states, “Sometimes lunging toward control trainer” 70 plus killer whale trainer accidents but no one talks about them.
The purpose of this lab was to explore and understand the scientific method and how to apply it to experimental procedures, as well as developing an understanding the importance of complete and concise presentation of experimental results obtained by statistical data analysis from collected raw findings. These learning objectives were accomplished by testing the emergence times of sponge creatures from their gelatin capsules by asking the question, “How does water temperature effect the time in which the sponge creatures materialize from their casing?”. An experiment was designed to test this using two different water temperatures and twenty four sponge gelatin capsules (1). Often referred to as “Magic Grow Capsules” or “Magic Animal Growing Capsules” these capsules are comprised of a sponge animal that has been placed in a small casing composed of gelatin. When placed in water the gelatin begins to break down.
When one thinks of a veterinarian, most associate this job title with treating dogs, cats, horses, cows, and other farm and companion animals. However, a relatively small field not known by the general population is that of marine veterinary medicine. If one has a passion for the water and a desire to help the creatures in it, marine veterinary medicine just may be the perfect career. Marine veterinarians are responsible for “conducting basic exams and evaluations, giving vaccinations, taking samples of blood and other bodily fluids, prescribing and distributing medication, evaluating behaviour, performing surgical procedures, and taking x-rays and sonograms” (Kramer “Life of an Aquatic Veterinarian”).
1.1 Abstract The purpose of quantitative analysis of protein using a spectrophotometer is to measure the concentration of proteins in a given sample. The experiment is conducted by laboratory method (Biuret Test) and using spectrophotometer to analyze the absorbance of reactants at 540 nm, hence determining the concentration of the proteins in a given sample. The purpose of stopped enzyme assay to study B-galactosidase is to determine the effect of temperature and concentrations of substrate on enzyme activity.
There are multiple points both at 43°C and at 72°C which indicates that liquid was collected at these temperatures. Based on this information, it would appear that two different liquids were present in solution and that one liquid has a boiling point of approximately 43°C and that the other has a boiling point of approximately 72°C. The literature value boiling point for DCM in is reported to be about 40°C and it is about 80°C for cyclohexane. Based on the graph, DCM was collected from 4 ml to 22 ml, thus 18 ml of DCM was collected.
What is meat tenderizer and what does it actually do? -It is an enzyme powder that contains papain and bromelain which breaks down the collagen in meats. 12. Design an experiment to test at what specific temperature the pineapple enzyme
Fix or replace damaged tissue; biology has always been the main concern for scientists. Today, the most important instrument for tissue engineering scientists to produce replacement tissues and implants to repair or replace damaged tissue. Tissue engineering is generating a new field of study in which the principles of engineering and biology to correct the damaged tissue, uses and can renewal, operation and maintenance of tissue healing. In order to use an ideal scaffold Tissue engineering should have features such as non-toxic Cell and tissue properties to be fit.
Cell viability assay: Introduction. Methods in Molecular Biology 740: 1-6. ThermoFisher Scientific. [Internet].