Stem Cells And Its Effects On The Field Of Bioengineering

Stem Cells And Its Effects On The Field Of Bioengineering
Stem cells has outgrown its medico–therapies in the field of bioengineering and also in the treatment of various conditions of the blood and immune
system, or to cure specific cancers Since it's a known fact that stem cells can different to specialized cells in an undifferentiated manner. As the paper
states the use of insulin producing cells from human or rat adipose tissue –derived stem cells transduced with pancreatic duodenal homeobox can could
cure type 1 diabetes. An alternative is, use of embryonic stem cells. Embryonic stem cells display the ability to differentiate in vitro into variety of cell
lineages. Insulin–secreting cells are derived from the mouse embryonic stem cells which can normalize blood glucose when transplanted ... Show more
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An alternative treatment to type 2 diabetes is insulin therapy since it is difficult to achieve desired glucose levels due to increased spread of the
disease. Transplantation of pancreas or replacement of patient's islets of Langerhans is an ongoing treatment capable of achieving normoglycemia.
However the limitations of immunesupression and scarcity of donor pancreas make this procedure not very desirable.
Stem cells therapy has been considerd as an alternative stratergy for replacement of damaged –cells , example embryonic stem cells , bone marrow
stem cells etc. Initially to cure type 1 diabetes , insulin–producing cells were created by transducing bone marrow stem cells with the pancreatic
duodenal homobox (Pdx1) gene. Although this experiment was successful in normalizing glucose levels in streptozotocin–induced diabetic mice, the
low level of insulin production and the unlikeliness of producing the culture conditions in vivo make this approach unfit for clinical applications.
Moreover, there was only a modest improvement in blood glucose levels by using experiments done by western blot, immunofluorscence and RT–PCR.
We need to further tune this treatment strategy to improve blood glucose levels to be able to beat the insulin therapy. A very efficient way to approach
this problem is using insulin–secreting cells derived
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The Health Of The Livestock Production Of Poultry Carrying...
The Acute phase proteins are produced as the develop of a response caused after a variety of conditions stimulated by inflammatory cells producing
chemical substances that activate specific organs to produce these proteins to reconstitute homeostasis and thus restore the injured tissue. Even though
the Acute Phase Response generated is not specific for each determined disease, the measurement of these proteins in an animal may indicate the
occurrence of an unhealthy condition, and a further diagnosis and prognosis of the illness could be accomplish. Regarding veterinary public health, the
livestock production represents a significant field from where products for human consumption are provided. Thus, the measurement of these proteins in
... Show more content on Helpwriting.net ...
These are plasma proteins which are released into the blood stream, mainly produced in the liver by the hepatocytes (Cray et al., 2009) as part of the
Acute Phase Response (APR), that either increase or decrease in response to the tissue damage (Jain et al., 2011; Stahl, 1987; Gruys et al., 2005). The
APR is a non–specific and complex systemic reaction, part of the early–defense or innate immune system, which is triggered by different stimuli such
as infections, trauma and inflammations in order to reconstitute homeostasis and induce healing (Cray et al., 2009; Tothova et al., 2014). As well as in
humans, APP in animals are sensitive to similar triggering events in response to inflammatory stimuli. A variety of APP have been recorded, such as:
fibrinogen, haptoglobin (HP), serum amyloid A (SAA); C–reactive protein (CRP); ceruloplasmin (CER), major acute phase protein (MAP) and О±1
acid glycoprotein (AGP) (Stahl, 1987; Gruys et al., 2005); differentiating their levels according to each species (Cray et al., 2009; Eckersall and Bell,
2010). Thus, APP have been described for their application in human medicine to diagnose and prognosticate different inflammatory and organ diseases.
However; altered levels of APP monitored in animals might provide an alternative method to measure conditions or situations that may be influencing
the concentration of these proteins in plasma (Tothova et al., 2014). For instance, in livestock; APP have been presented as markers of health

Gfba Gene Essay
Effect of cultivation conditions and gfba gene on formation of biofilm by Streptococcus dysgalactiae
Abstract
The pathogenesis of Streptococcus dysgalactiae is attributed to a combination of extracellular factors and properties such as adherence and biofilm
formation. The aim of this work was to evaluate the influence of different factors, additives and bovine milk compounds on S. dysgalactiae biofilm
formation, as the presence of the gfba gene by PCR. Additionally, extracellular DNA and the effect of DNaseI were evaluated in the biofilms yielded.
Optimal biofilm development was observed when the pH was adjusted to 7.0 at 37 ВєC. Additives as glucose and lactose reduced biofilm formation as
bovine milk compounds tested. PCR assay showed ... Show more content on Helpwriting.net ...
Treatment of the disease is currently based on antimicrobial susceptibility tests according to Clinical and Laboratory Standards Institute (CLSI) [5].
Standard therapy designed for bacteria isolates at planktonic state continues to be applied. However, the pathogenic profile and the antimicrobial
resistance are totally different from biofilms, causing reduced cure rates. Furthermore, the ability to form biofilm is important both from the
pathogenicity to the animal as for the manufacturing milk industries, where the pathogen can adhere to abiotic food processing structures and persist in
adverse conditions through biofilms. The development and establishment of the biofilm depend on the ability of pathogen to adhere to bovine
mammary epithelial cells. S. dysgalactiae have many virulence factors including the capacity to bind to the host's cells surface by a protein called
GFBA, which is involved in adherence and internalization, by its linking to lactoferrin [6]. It was demonstrated that the S. dysgalactiae isolated from
mastitis cows was able to produce biofilm [7]. Economic negative effects in the milk production around the world and the possible involvement of
biofilms in S. dysgalactiae mastitis infections, address the research to the study of factors that may contribute for the biofilm production in order to
establish more effective treatment strategies. Different studies report that environmental conditions influence the capacity to form biofilm in

Hi-Res Standard Agae Dyes
2.3.0 DNA Quality For each individual DNA sample 2Вµl of DNA, 8Вµl of deionised water and 2Вµl of 6x blue/orange loading dye (ThermoFisher)
was added to a microcentrifuge tube and mixed using the pipet. The DNA solutions were then refrigerated while a 2% agarose gel was made. Initially
2g of Hi–Res standard agarose (Bioproducts Ltd. Yorkshire, UK) was weighted out on a balance (Denver Instrument S–203) and added to a 250ml
conical flask. Once this was completed 98ml of deionised water was added and heated on a hot plate (Halogen Hotplate–HT2). The flask was
continuously heated and stirred until the agarose powder had fully dissolved. Then the solution was cooled by running the base of the conical flask
under water while stirring. Once the solution had begun to cool 2ml of 50X Tris–acetate–EDTA (TAE) (ThermoFisher) and 10Вµl of Gel Red Nucleic
Acid Stain 10,000X in DMSO (Biotium. Fremont, USA) via pipetting. Once mixed the solution was poured into gel electrophoreses tank including
well comb and left to set for 30 minutes. Once set the gel is immerged in approximately 300ml of 1x TAE, then 10Вµl ofDNA solutions including a
PCR sizer 100bp DNA ladder (Norgen Biotek Corporation. Thorold, Canada) ... Show more content on Helpwriting.net ...
Buckinghamshire, UK) were used within each supplied tube. Each bead contains the desired ingredients needed for the successful PCR reaction,
stabilizers, bovine serum albumin to increase overall yield, four deoxynucleotides triphosphates (dNPT), adenine, guanine, cytosine, tyramine ATP,
approximately 2.5 units of puReTaq DNA polymerase and a reaction buffer. When reconstituted to the final volume of 22Вµl, the final volume of each
dNTP is 176ВµM in 8.8mM of Tris–HCL which at room temperature has a pH of 9.0. Finally, 44mM of potassium chloride and 0.66mM of
magnesium chloride. All PCR was preformed within a fume cupboard (Bassaire 06HB) to minimise the risk of either moisture or genomic

Glorimetric Blocking Solution Essay
пѓ Coating solution (pH = 9.6): 1.59 g Na2CO3,2.93 g NaHCO3, add sterile deionized water (ddH2O) and dilute to 1000 mL, autoclave spare .
пѓ Washing buffer solution (pH = 7.4): 8.0 g NaCl, 0.2 g KCl, 0.2 g KH2PO4, 3.58 g Na2HPO4 12H2O, 0.5 mL Tween–20, plus ddH2O volume to
1000 mL.
пѓ Blocking solution: 1 g of bovine serum albumin (BSA), plus washing liquid volume to 100 mL or 0.1 mol citric acid solution: 2.1 g of citric acid,
plus ddH2O volume to 100 mL.
пѓ Substrate buffer (pH = 5.0): 25.7 mL 0.2 mol / L Na2HPO4, 24.3 mL 0.1 mol / L citric acid solution , 50 mL ddH2O.
пѓ Stop Solution to 2 mol / L sulfuric acid: 22.2 mL of concentrated sulfuric acid, plus ddH2O volume to 200 mL.
(1) The package was suppressed kisspeptin–54 antigen: each well of 100 ng / 100 ОјL with leaving three empty holes blank. The plate was sealed
using sealing plate incubated at 4 В° C ... Show more content on Helpwriting.net ...
An appropriate step has been accordance to kit instructions to determine the concentration range, intra–assay precision and cross–reactivity with other
related proteins.
Assay Procedure
1. Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples were added in duplicate to the Microtiter plate.
2. Add 50Ојl of Standard or Sample to the appropriate wells. Blank well did not receive anything.
3. Add 100Ојl of enzyme conjugate to standard wells and sample wells except for the blank well cover with an adhesive strip and incubated for 60
minutes at 37В°C.
4. Wash the Microtiter Plate 4 times.
Aspirate all wells, then wash plates four times using Wash Buffer (1X) and set fill volume at 350 ОјL/well/wash. After the final wash, invert plate and
blot dry by hitting plate onto absorbent paper or paper towels until no moisture

Standard Crive Lab Report
Purpose:
1. Assay a series of dilutions of known concentration and prepare the standard curve.
2. Determine the concentration of unknown 1 and unknown 2 based on the standard curve.
Methods:
I. Preparation of Diluted Bovine Serum Albumin (BSA) Standards
Five 1.5–ml tubes were prepared and labeled as 1 – 5. Guided by Table 1, agents were added in order. Respectively pipetted 325 Ојl of BSA Stock and
325 Ојl PBS into tube 1. Mixed the solution by pipetting. Pipetted 325 Ојl of tube 1 solution into tube 2, and added 325 Ојl of PBS into tube 2. Mixed
the solution by pipetting. Pipetted 325 Ојl of tube 2 solution into tube 3, and added 325 Ојl of PBS into tube 3. Mixed the solution by pipetting.
Pipetted 325 Ојl of tube 3 solution into tube 4, ... Show more content on Helpwriting.net ...
The coefficient of variation was expressed as a percentage.
Conclusions:
The standard curve is used to determine the concentration of each unknown sample. The trendline in Figure 1 has a desired regression value: R2 = 1.
R2 measures the distance between the fitted line and the data points. The value ranges from 0 to 1, and the higher it is, the better the model fits the
data points. The value 1 suggests that the model is the best–fit model in which all the variability of data around their mean can be explained. Therefore,
the regression value is acceptable and the standard curve is reliable.
For each unknown sample, the SD value and CV percentage is low.
Applications:
The BCA method can be applied to many research areas. During the assessment on the caries–associated virulence (production of acid, extracellular
polysaccharides, proteins and metabolic activity) biofilms from Candida

Membranes Case Study
GO has also been mixed with the polymeric matrix to fabricate different nanocomposite membranes, such as polyvinylidene fluoride (PVDF) MF/UF
[141,224,225], polyether sulfone (PES) NF membranes [226] and PSf [162]. One of the main drawbacks of the blending technique is the absence of
GO direct exposure to foulants, which restricts the interaction between foulants and GO. Accordingly, the superior GO features, in terms of antifouling,
will not be entirely exploited. The direct deposition of GO nanosheets on the membrane surface is a more effective technique for fouling control, hence
the GO/foulants interaction can be maximized. The exceptional structure of GO confers a considerably greater contact area for the convenient
interaction... Show more content on Helpwriting.net ...
The employment of GO leads to the production of small, uniform, stable and distributed AgNPs in addition to improved silver loading and ion
release control. Several applicable and effective techniques for the fabrication of freestanding ultrathin rGO membranes were reported. One of these
techniques was exemplified by Liu et al [220], in which a reinforced GO membrane was shaped on a mixed cellulose ester (MCE) filter through the
filteration of diluted GO dispersion. The formed GO/MCE membrane was then positioned over a hydriodic acid solution, where the GO membrane
rapidly changed to a black colour, confirming the reduction of GO to rGO by the acid vapour. Furthermore, a new water filtration membrane was
fabricated by LbL–SA as it is an effective method to generate dense layers on a membrane support [229]. This method was successfully used to
integrate the negatively charged GO nanosheets with a porous poly(acrylonitrile) (PAN) support through the positively charged poly (allylamine
hydrochloride) (PAH) by electrostatic interaction [211]. Additionally, GO
–based FO membranes were fabricated in several studies. The first trial was
conducted by Park et al [222], in which GO nanosheets were combined with the PSf to produce a novel membrane with a PSf/GO composite support
layer. The PA active layer was then shaped on the PSf/GO via IP to obtain the

What Are Gel Electrophoresis?
Gel electrophoresis is a gel technique that separates DNA and proteins based on their mass, by means of an applied electrical field that passes
through an agarose or polyacrylamide gel. The concentration of agarose in the gel is commonly 0.8% to 1.0%, since agarose is expensive. The
gel is embedded on a buffer, pH of 8.3, which keeps the pH of the solution at equilibrium. Assuming that at typical pH, DNA is negatively charged,
denatured protein samples are placed in the wells located on the negative electrode side; hence the positive electrode side is located at the other
end of the gel. As the positive and negative electrode sides are connected to a power source, protein sample migrates to the positive side of the gel.
Migration of proteins is not necessarily based on their mass or mass to charge ratio; protein migration across the gel is based on their size. In other
words smaller molecules will travels further than bigger molecules. Since the SDS gel contains agarose or polyacrilamide, molecules have to be small
enough to migrate to the other end of the gel without getting stuck on the way. Protein samples separated by size via SDS–Page can be identified using
mass spectroscopy; which will require proteins from the gel to be treated using trypsin; which is an enzyme that digests proteins. Assuming that
samples were treated and storaged properly, the digestion of peptides begins with the rehydration of proteins using trypsin, followed by the incubation
of proteins using a

Cell Cycle Analysis Paper
Cell cycle analysis. PC3 (8Г—104 cells/well) and LNCaP (3Г—105 cells/well) cells were seeded in 6–well plates in 1.5 mL of complete growth
media. Cells were treated with 2.5 ОјM of free sorafenib or an equivalent sorafenib concentration of SMA–Sor, 3 ОјM of free nilotinib or an equivalent
nilotinib concentration of SMA–Nilo, DMSO or SMA for 48 h. Cell cycle distribution was assessed using propidium iodide staining, as previously
described {Somers–Edgar, 2011 #82}. Samples were analysed using a FACScalibur flow cytometer (BD Biosciences, San Jose, CA, USA) and the
proportion of cells in each of G0/G1–, S– and G2/M–phases were determined using CellQuest Pro software (BD Biosciences, San Jose, CA, USA).
Apoptosis analysis. PC3 (8Г—104 cells/well)... Show more content on Helpwriting.net ...
The cells were washed with PBS and then incubated in serum–free media and treated with 2.5 ОјM of free sorafenib or an equivalent sorafenib
concentration of SMA–Sor, 3 ОјM of free nilotinib or an equivalent nilotinib concentration of SMA–Nilo, DMSO or SMA for 48 h. Following
treatment, media was collected, centrifuged to remove cell debris, and freeze–dried for 12 h. Samples were rehydrated and mixed with loading buffer
(0.4 M Tris, pH 6.8, 5% SDS, 20% glycerol, 0.03% bromophenol blue). For zymography, samples were loaded on a 10% SDS–polyacrylamide gel
containing 1 mg/mL of gelatin. After electrophoresis, the gels were incubated in renaturing solution (2.5% Triton–X–100 (w/v)) for 30 min at room
temperature and then for 24 h at 37В°C in a developing buffer containing 50 mM Tris, pH 7.5, 200 mM NaCl, 4 mM CaCl2, and 0.02% NP40. The
gels were then stained with Coomassie blue R250, and regions without staining were indicative of gelatin lysis. The gels were briefly rinsed and
scanned. For MMP–9 and isthmin–1 secretion, samples were loaded on a 10% SDS–polyacrylamide gel, and the expression of MMP–9 and isthmin–1
assessed by western blotting using anti–MMP–9 (D6O3H, Cell Signaling) and anti–isminth–1 antibodies (Biorbyt, San Francisco, CA,

The Cardiovascular System And Its Blood Transporters Essay
Introduction: The cardiovascular system and its blood transporters continue to amaze scientists, as wells, as others due to its fascinating functions. The
cardiovascular system inhales oxygen and functions to pump blood. It also, transports nutrients throughout the body. This system, then, administers a
crucial chemical reaction between the blood and hemoglobin. Hemoglobin is a particular protein responsible for binding to oxygen molecules and
transporting those molecules to the body's tissues. Because of hemoglobin's important function, the body is able to properly distribute oxygen
molecules, proteins, and hormones throughout the whole body.
A spectrophotometer becomes a useful tool in determining the concentration and the identity of an unknown sample. The data gathered from a
spectroscope measures absorbance through light waves. A spectrometer measures how much a sample absorbs light as light passes through the sample
inside of the device. Also, spectronomy is a quantitative analysis, which absorbs light over a variety of wavelengths. It depicts the movement of
electrons. More concentrated solutions will possess a higher absorbance due to the electrons being excited and moving into higher energy levels. The
principle of spectrometry can be applied when determining the concentration of an unknown sample as well. From the information gathered, one can
create a standard curve in order to discover the range of absorbance and the identity of the unknown sample.
A standard

Lab Report : Bca Protein Assay
Shrutika Mintri/ E2Date : 09/17/2014 Lab Report #1 Project Title: BCA Protein Assay Purpose: BCA (Bicinchoninic acid) Assay otherwise known as
the "Smith Assay" has the fundamental purpose of determining the protein concentration of the two given unknown proteins in the sample solutions.
The absorbance is measured using a Plate reader and a Standard curve is generated. Also, the different types of pipetting techniques are assessed in this
Assay. Methods: (1) There are two methods using which the BCA Protein Assay can be performed
– Test tube method and Micro plate method. The
following experiment has been performed using the Microplate procedure. VialVolume of Diluent Volume and Source of BSAFinal BSA Concentration
A325 ОјL325ОјL of Stock1000Ојg/ml B325ОјL325ОјL of vial A dilution500Ојg/ml C325ОјL325ОјL of vial B dilution250Ојg/ml D325ОјL325ОјL of
vial C dilution125Ојg/ml E400ОјL00Ојg/ml= Blank 1.Preparation of Diluted Albumin (BSA) Standards: The following Diluted Albumin (BSA)
Standards have been prepared using Bovine Serum Albumin (BSA) Standard Solution (2.0mg/ml) and the Diluent Phosphate Buffered Saline (PBS)
having

Ligand Bonding Lab Report
For this experiment, ligand binding was introduce into this course. What is ligand binding? It is a smaller molecule that bind to a larger molecule,
called a substrate (Berger). Bovine serum albumin (BSA) is a protein that derive from cow that has a large molecular weight up to 66,436 Da. Phenol
red dye has a molecular weight of 354.88 Da, and is binding to bovine serum albumin at pH 4, pH 6, pH 8, and pH 10, and our target was to determine
the optimal pH. The optimal pH ranging between 3 to 5, and that it will appear to have two peaks on the graph. The first peak would be the phenol
red–BSA at maximum and the second peak is only phenol red at maximum because it does not bind to BSA. After gather each pH peaks, then calculate
the percent bound.

Analysis Lab Report Sample
Table 1 OD of E.coli Cell Cultures Compared to Blank Samples7 Table 2 The Averages Absorbance of Bovine Serum Albumin In E.coli Induction
Experiment7 Table 3 The Averages Absorbance of The Cell Lysate & Cell Pellet In E.coli Induction Experiment7 Table 4 The Concentration of The
Cell Lysate & Cell Pellet In E.coli Induction Experiment8 Table 5 Calculation of SDS Standard Curve for E.coli Induction Experiment16 Table 6 The
Calculated Mw of PTEN in E.coli Induction Experiment8 Table 7: The Averages Absorbance of All Samples Proceed Along With Affinity
Chromatography Experiment16 Table 8 The Calculated Concentration of Affinity Chromatography Experiment9 Table 9 The Calculation of
SDS–PAGE Standard Curve for Affinity Chromatography Experiment17 Table 10 The Mw of PTEN In Affinity Chromatography Experiment10 Table
11 The Calculation of Mammalian Cell Density10... Show more content on Helpwriting.net ...
Table of Figures Figure 1 Order of The Samples Loaded to SDS–PAGE for E.coli Induction Experiment8 Figure 2 SDS–PAGE for E.coli Induction
Experiment8 Figure 3 The Samples Order of Affinity Chromotography Expermint9 Figure 4 SDS– PAGE of Affinity Chromotography Expermint9
Figure 5 The Order of Western Blotting Samples11 Figure 6 The PVDF Membrane Showing The ladder after Westren Blotting11 Figure 7 The
Chemiluminscence Reaction on PVDF Membrane11 Table of Graphs Graph 1 Bradford Assay Standard Curve for E.coli Induction7 Graph 2 SDS
Standard Curve for E.coli Induction Experiment16 Graph 3 Affinity Chromatography of His6 Fused PTEN9 Graph 4 The SDS–PAGE Standard Curve
for Affinity Chromatography Experiment

The Bacterial Culture And Growth
Results
Bacterial culture and Growth
Isolates were resuscitated from storage by culturing on glucose yeast extract agar and single colonies of the isolates inoculated on glucose yeast
extract broth incubated at 28oC in a rotatory shaker at 180 rpm showed that growth saturation at 0.8 O.D was after 24 hr of incubation showed that
Rhodococcus species are aerobic
Biochemical Assay
Determination of concentration of crude extract was based on Bradford Coomassie brilliant blue assay which forms a complex between the protein dye
and protein in solution which causes a shift in the maximum absorption of light that passes the dye from 465 to 595 nm (Bradford, 1976). The amount
of light that passes through the protein solution is ... Show more content on Helpwriting.net ...
Fig5: Standard curve diagram of the concentration of BSA measured at an absorbance of 595 nm.
The volume of each sample for restriction digest was calculated using the formula; C1V1 = C2V2
Where C1 = Initial Concentration V2 = Initial Volume C2 = Final Concentration V2 = Final Volume
Table4: Concentration of crude extracts
Isolates Concentration of crude extracts (Вµl)
A
1.0
B 1.7
C1.4
D0.7
E0.8

F1.5
G1.0
H0.8
I0.7
J0.4
K1.5
L0.9
M1.2
N1.1
O1.0
P0.8
Q1.5
R1.0
S1.5
T1.2
U1.7
V1.2
W0.9
X1.6
Y1.1
Z1.6
Electrophoresis Analysis
Agarose gel electrophoresis used for the separation of the fragments was based on the migration of the negatively charged DNA to the positively
charged anode. DNA has a uniform mass/charge ratio therefore allowing the molecules to be separated by size within an agarose gel such that the
distance travelled is inversely proportional to its molecular weight. The rate of migration is controlled by the following; size of DNA molecule,
agarose concentration, DNA conformation, voltage applied presence of ethidium bromide, type of agarose and electrophoresis buffer.
Figure 6 present

The Infection Of Cattle And Dogs
The genus Babesia is composed of tick–borne intraerythrocytic apicomplexan protozoans that cause babesiosis in mammals, including humans
(Schnittger et al., 2012). The infection of cattle with Babesia bovis causes massive economic losses in the livestock industry worldwide (Bock et al.,
2004; Malak et al., 2012).
Similarly, Babesia gibsoni infects dogs worldwide, causing serious concern among pet owners with regard to animal health and welfare (Trotta et al.,
2009; Yeagley et al., 2009). Cattle and dogs clinically infected with B. bovis and B. gibsoni manifest fever, anemia jaundice, hemoglobinuria,
hypertensive shock, and more often deaths if left untreated (Bock et al., 2004). Due to the limited range of anti–Babesia drugs available, the treatment
of babesiosis is becoming increasingly difficult, particularly in areas where the disease is endemic (Gohil et al., 2012; Lin et al., 2012). Therefore, it
is clear that alternative strategies are required for the discovery and development of the much needed anti–Babesia drugs.
The M17 leucine aminopeptidases of Plasmodium falciparum
(PfM17LAP) and Plasmodium vivax (PvM17LAP) have been characterized and validated as a drug target (Cunningham et al.,
2008; Lee et al., 2010). The malaria parasite, just like Babesia species, destroys erythrocytes, causing many of the clinical symptoms responsible for the
disease. The PfM17LAP is involved in host cell hemoglobin digestion, metabolic regulation, intracellular housekeeping, and antigen

Bradford Assay On Unknown Concentrations Of Proteins
Bradford Assay on Unknown Concentrations of Proteins
Taylor Coleman
September 27, 2016
Lab Group 3
BIOL 1111: General Biology Lab
Fall 2015
Section 107
Chad Perry
Abstract Colorimetric assay is a process of determining a concentration of a solution based on absorbance of light. The purpose of this lab is to
determine if the Bradford assay is an accurate way to determine an unknown concentration of two samples of protein. The Bradford assay is done by
measuring wavelength of light passing through a cuvette filled with Bradford dye and concentrations of PBS and proteins. After the cuvettes are mixed
they are placed into a spectrophotometer to measure wavelength. The wavelength given will be used to plot a standard curve based on concentration
(x–axis) and wavelength (y–axis). The standard curve is then used to measure an educated guess on the concentrations of unknown protein
concentrations. We hypothesized that if we use the Bradford assay and colorimetric spectrophotometry we can determine an accurate concentration of
two unknown concentrations of proteins. The results of this lab failed to reject our hypothesis based on accurate measurements of protein
concentrations. The standard curves are drawn with a linear increasing slope. The Bradford assay is an accurate way to demine the concentration of an
unknown concentration.
Introduction Colorimetric assay is a process determining the concentration of a chemical element or compound in a solution

Sds Page (Sodium Dodecyl Sulfate Polyacrylamide Gel...
Gel electrophoresis is a routine laboratory procedure in biochemical studies that takes advantage of a protein's amphoteric nature to determine its
molecular weight and charge by running the sample through a gel matrix under the influence of an electrically charged field. A popular example of gel
electrophoresis is Sodium dodecyl sulfate–polyacrylamide electrophoresis or SDS– PAGE which has been used in this experiment to supposedly
determine albumin and casein's molecular weights respectively. The system used in gel electrophoresis consists of 3 major parts: a stationary phase, a
mobile phase) and an electrical power supply.
In SDS–PAGE the polyacrylamide gel serves as the stationary phase, the buffer (Tris–HCl in this case) as the ... Show more content on Helpwriting.net
...
On the other hand, the Tris–HCl buffer and deionized water provides the system its shape since the polyacrylamide complex is just a thin film of
interlocking polymer fibers without any liquid to trap within.
Polyacrylamide gels are usually preferred over other gel types such as agarose in protein electrophoresis because of various benefits such as its
ability to be cast in various pore sizes which provides options for the researchers who observe protein samples with various Kda's depending on how
separated they aim the bands to be or how well the sample is sieved. It is also preferred because of its charge neutrality upon setting as it does not
interfere with the electrophoresis which is highly reliant on the sample and the surrounding's charges within the system. Lastly, the gel itself is
colorless and protein stains do not adhere to the gel so there will be minimal problems with unnecessary stains that aren't from the staining solution.
APS and TEMED was used to initiate the binding of acrylamide to bisacrylamide via a vinyl addition reaction where TEMED catalyzes the
decomposition of APS into free sulfide molecules which then initiates the formation of the polyacrylamide complex.
Sodium dodecyl–sulfate or SDS is a protein denaturant that destroys weak interactions within the protein molecule which unfolds it

Spectrophotometer: Concentration and Standard Curve Essay
Lab Report 4
Biology 103
Prof. Dr. Johnson
Spectrophotometer; the finding of protein concentration of an unknown sample of BSA, and by using the standard curve.
Abstract A spectrophotometer's purpose is to use colors of the light spectrum to determine the concentration of light absorbing molecules in a solution.
(p.59) In this particular lab, our mission was to determine the protein concentration and the standard curve of the unknown sample of BSA. This, by
preparing five dilutions of the unknown solution of BSA together with other known concentrations, and then experimenting by observing how the
concentrations were passed through the spectrophotometer. The outcome resolved in the absorption levels being decreased, and this... Show more
The same solution of 0.5 ml BSA was then added from test tube 1 to the test tube 2 after being properly mixed, and from test tube 2 the solution was
being added to test tube 3, and so forth all the way up to test tube 5, with the same exact procedure. From the last tube, we then disposed the 0.5 ml
solution. After above procedures, we now labeled another test tube "blank"; 0.5 ml blank distilled water was purred into the tube with the serial dilution
of 1:10. We also had a tube C labeled "unknown" with the same 0.5 ml of solution. And after adding 5ml of Coomassie Blue to each tube (1–5) and to
the blank, the result of absorbance was read at 595 nm.
Result | Protein Concentration(Вµg/ml)| Absorbance(595 nm)| Tube 1| 120| .922| Tube 2| 60| .557| Tube 3| 30| .377| Tube 4| 15| .180| Tube 5| 7.5|
.082| Unknown: .759 |
The ending result of this experiment confirms that as five test tubes are lined up with the varying level of absorbance, different results in the level of
absorbance will appear as well, this is visible in above table. Thus, this is due to the varying amount of water in the solution. The blank sample had a
0.30 in its level of absorbance.
Calculating serial dilutions and their concentrations
(Examples of test tube 1 and test tube 2for respective dilution and concentration)
Dilution or D = previous dilution (if any) Г—the

Should Vaccinations Be A Choice?
Should Vaccinations Be a Choice? According to scientists, a vaccines dominant purpose is to customize immune systems and to allow them to work in
contradiction of diverse viruses and infections. However, in place of assisting the struggle against infection and certain diseases, vaccines can be the
key cause of infections and illness (Sepkowitz, 2012). Therefore, vaccinations should be an individual choice and not a requirement by state and school
legislation. The first vaccination invented was for smallpox in 1798 by a man named, Edward Jenner (Kotz, 2009). His work was herald as the leading
scientific effort to control a transmittable virus by the measured consumption of a vaccination. Today, current medication is founded on Pasteur's germ
philosophy of infection (Link, 2005). Scientists now have vaccinations for almost everything from vitamin vaccinations to vaccines preventing
illnesses. Oklahoma school and state legislation requires a variety of vaccinations and require them at various ages from as early as three months
through eighteen years of age. Some of these vaccinations come with severe side effects if any side effects at all. Also these vaccinations come with a
possibility of causing the infection that the vaccine was made to prevent. This is when the situation of an individual's immune system comes into play
whether to accept or reject the vaccination. Vaccinations that are required by the state and school legislation of Oklahoma are; DTaP (diphtheria,

The Spectrophotometer Determination of Protein...
The Spectrophotometer Determination Of Protein Concentrations And The Effects Sodium Dodecyl Sulphate And Triton X
–100 Have On Protein
Concentration.
INTRODUCTION
Spectroscopy is used as a collective term for all the analytical techniques based on the interaction of light and matter. Spectrophotometry is one of the
branches of spectroscopy where we measure the absorption of light by molecules that are in a gas or vapour state or dissolved molecules/ions (Tombs,
et.al, 1959). Spectroscopy is the use of electromagnetic radiation by a sample in order to identify components of the sample (qualitative analysis) or to
measure the amount of material in that sample (quantitative analysis) (Tombs, et.al, 1959). Absorption is a process in which... Show more content on
Helpwriting.net ...
MATERIALS AND METHODS
The BIORAD dye was firstly prepared by dissolving 0.225g of Coomassie Brilliant Blue G–250 (Merck, South Africa), in 67.5ml of Methanol
(Riedel–de–HaС‘n, Germany) and 291ml of 85% (w/v) Phosphoric Acid, diluted to 450ml with dH2O. Six test tubes were labelled SC1–6 and the
0.1mg/ml stock of Bovine Serum Albumin (Roche Diagnostics, Germany) and dH2O was pipette into their respective test tubes according to Table 1.
Table 1: Table of the BSA and dH2O volumes to be pipette and placed in the spectrophotometer to obtain absorbencies for the standard curve
Test Tube NumberTotal Volume (О
јl)Protein (BSA) Stock (Ојl)H2O (Ојl)Amount of Protein (Ојg)
12400024000
224005023505
32400100230010
42400150225015
52400200220020
62400250215025
600Ојl of the BIORAD dye agent was pipette into each test tube making up the volume to 3000Ојl. All the test tubes were then covered with Parafilm
and placed in the Vortex (Lab Pro cc) for a few seconds and they were incubated at room temperature for 5–60 minutes. Simultaneously another set of

test tubes were labelled X1–8 and pipette with 0.1mg/ml Lysozyme stock (Merck, Germany), BSA, 1% (w/v) Sodium Dodecyl Sulphate prepared with
dH2O, 1% (v/v)

Synovial Bacterial Lab
Edwanny Nivar
Professor Fair
Microbial Physiology and Genetics
12 November 2017
Role of phenol–soluble modulins in formation of Staphylococcus aureus biofilms in synovial fluid
ABSTRACT/OVERVIEW QUESTIONS:
1.Who did the research, and where?
a.This research was done by Sana S. Dastgheyb, Amer E. Villaruz, Katherine Y. Le, Vee Y. Tan, Anthony C. Duong, Som S. Chatterjee, Gordon Y. C.
Cheung, Hwang–Soo Joo, Noree J. Hickok and Michael Otto.
b.It was conducted at the Pathogen Molecular Genetics Section, Laboratory of Human Bacterial Pathogenesis, National Institute of Allergy and
Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA; and at the Department of Orthopedic Surgery, Thomas Jefferson
University, ... Show more content on Helpwriting.net ...
a.Staphylococcus aureus is a major cause of septic arthritis and orthopedic infection, in particular those developing on prosthetic joints after arthroplasty.
Joint infections can cause prolonged disability and increased health care cost, due to prolonged antibiotic treatments, multiple surgeries, and in some
cases joint fusion. If there is rejection of treatment, these infections can cause significant morbidity, including loss of limbs, systemic infection, and
even death.
b.Biofilms are surface–attached bacterial agglomerations that frequently develop on indwelling medical devices.
c.The matrix that connects cells in a biofilm consists of a variety of chemically different macromolecules, such as polymeric proteins, teichoic acids,
extracellular DNA and polysaccharides.
d.The pronounced recalcitrance of S. aureus joint infections to antibiotic treatment is due to exceptionally strong bacterial aggregation and biofilm
formation, which renders even high concentrations of antibiotics given to patients ineffective.
METHODS, RESULTS, AND CONCLUSIONS
4.How was the research conducted and with what materials and methods (experimentally or otherwise), and why were these methods used?
a.The research team started by draining human synovial fluid (SF) from the joint during total knee arthroplasty in the operating room and collected this

with permission of the Thomas Jefferson University Institutional

Assay For Quantification Of Anti Rabbit Proteins
9/30/2014
Rakshitha. P. Miskin
01399629
Tuesday– 10
Lab #2
MicroELISA assay for quantification of anti– rabbit protein antibodies in mouse before and after immunization
Objective
Preparation of microELISA plates for the detection of antibodies in sera from pre–immunized mouse against rabbit protein and comparing this to sera
extracted at weeks 1 and 2 after immunization and re–immunization of the mouse to check for increased titer of antibodies in the sera
Materials and methods
Preparation of phosphate buffer saline (PBS): A tablet of PBS sigma #P4417 was added to 200 mL of water maintaining pH between 7.2–7.4.
Preparation of PBS/BSA: 10mL of 1% bovine serum albumin (BSA) in PBS was prepared (0.1mg BSA in 10mL PBS) and the solution... Show more
The strips were incubated for 30 minutes at room temperature and then washed 3 times with PBS–tween. To each of the wells, 100ВµL of peroxidase
conjugated goat anti– mouse immunoglobulin was added and incubated at room temperature for 1 hour. The strips were washed again with PBS–
tween and then with PBS. 100ВµL each of activated ABTS (100Вµl hydrogen peroxide was added to 1mL 2, 2'–azino–bis
(3–ethylbenzothiazoline–6–sulphonic acid) (ABTS)) was added to each of the wells. The strips were incubated for a few minutes and the absorbance
was read at 405nm in a microELISA plate reader.
Calculations
Dilutions for strip 1:
Well #SeraDilutionVolume of sera Volume of PBS
–tweenTotal volume
1–Negative control–100ВµL100 ВµL
2–Negative control–100 ВµL100 ВµL
3Pre–immunized sera1:1022 ВµL198 ВµL220 ВµL
4Week 1 immunized sera1:1020 ВµL180 ВµL200 ВµL
5, 61:10025 ВµL (of 1:10 stock)225 ВµL250 ВµL
7Positive control100 ВµL–100 ВµL

8Positive control100 ВµL–100 ВµL
Dilutions for strip 2:
Well #SeraDilutionVolume of sera Volume of PBS
–tweenTotal volume
1–Negative control–100ВµL100 ВµL
2–Negative control–100 ВµL100 ВµL
3Pre–immunized sera1:1022 ВµL198 ВµL220 ВµL
4Week 2 immunized sera1:10 (Stock 1)15 ВµL135 ВµL150 ВµL
5, 1:100 (Stock 2)15 ВµL (of stock 1)135 ВµL

Microcapsules Essay
Layer–by–Layer Approach for Making Microcapsules
Muhammad Faiz Omar (201308264)
4th December 2014
Abstract
The fabrication of hollow multilayer microcapsules in the beginning is initiated by layer–by–layer (LbL) self–assembly technique of oppositely
charged polyelectrolyte on colloidal template followed by core decomposition. The main concept of microencapsulation is to protect the contents of
fabricated microcapsules from the harmful environment in which it is surrounded. In this literature review, I shall outline the technique of microcapsules
encapsulation employing LbL approach, lists of polyelectrolytes and templates and application of microcapsules.
Introduction
Over the past decades, nanometre–sized or known as microcapsules have been benefited the approach of Layer–by–Layer(LbL) technique due to the
ability of tailoring their properties(e.g., size , shape, colloidal stability, surface–functionality, composition) that are biocompatible and
biodegradable.1–3,10They are broadly used in many applications including food technologies, cosmetics, textiles industry and pharmaceuticals. The
LBL assembly of multilayers are fundamentally built up by the alternating deposition of oppositely charged polyelectrolytes onto a sacrificial template
such that the charge oscillates between positive and negative with each layer.1,4 The establishment of nanocoatings using LbL assembly method
distinguishes itself in its intelligibility compared to the more conventional method. The

Paper Chromatography Lab Report
Paper Chromatography and Spectrophotometry Used to Identify Amino Acids and Concentration of Protein in Solution
Introduction: Paper chromatography was used to characterize known amino acids in solution with an accordant Rf value. These Rf values were then
used to identify a solution with unknown amino acids and observed Rf values. An Rf value is calculated using the following formula:Distance
Traveled by solute (cm)Distance Traveled by solvent (cm), where the distance traveled by solute is measured from the origin line to the center of the
solute spot (Lombard, p19). A polar matrix, composed of cellulose, was utilized in tandem with a nonpolar solvent (Ammonia:Ethanol:dH2O) during
paper chromatography (Freeman, et al. p113). This diversity ... Show more content on Helpwriting.net ...
Unknown Y had two observed Rf values of 0.305 and 0.634. The primary Rf value deviated only 0.001 from the calculated Rf of Histidine while
the secondary amino acid deviated 0.036 from the nearest Rf value of 0.598 for Valine. The percent error for this set of Rf values is only 6.02%,
which we find to be within reasonable ranges. Despite the more significant error present in the secondary amino acid identification, Unknown Y is
still believed to be a solution of Histidine and Valine. Unknown X had two observed Rf values of 0.147 and 0.529; of which, only the latter
devotes form a calculated amino acid Rf value. The former Rf matches exactly to Aspartic Acid while the latter deviates 0.005 from the accepted
Proline value. Given this close adherence to calculated values, Unknown X is hypothesized to be a solution of Aspartic Acid and Proline. This is
concurrent with the stated hypothesis as charged amino acids such as aspartic Acid, Histidine, and Lysine had Rf values ranging from 0.147 to
0.306 while those acids with nonpolar tendencies reported values from 0.524 to 0.776 (Freeman, et al. p80). The average value of polar/charged
amino acids was 0.228, which was 0.405 less than the average value of non polar groups 0.633. Clearly, the nonpolar amino acids were interacting
with the matrix in a way that decreased their travel distance over 1 hour, therein supporting our hypothesis that adsorptive behavior will result in an
increase Rf value relative to amino acids with polar or charged R–groups. The second procedure performed was two parts wherein we both optimized
the wavelength for spectrophotometry and established a standard curve. We established an absorption spectrum based on the incremental increase in
wavelength by 10 nm from 490 nm to 570 nm

Physioex 12 Answers
Review Sheet
Exercise 12
Serological Testing
NAMEKali Rothfuss
LAB DATE/TIME6/22/11
Positive and Negative Controls
1.Why are there a number of washing steps in serological tests? The are a number of steps needed in order to remove any non specific binding that
may have occurred.
2.Describe how you would know that you had a "false positive" result. What does this mean for the rest of your results?
A positive result with a negative control indicates a "false positive" and your results are invalidated.
3.Describe how you would know that you had a "false negative" result. What does this mean for the rest of your results?
A negative results with a positive control indicates a "false negative" and your ... Show more content on Helpwriting.net ...
19.If a patient tests positive for HIV using the ELISA test but negative using Western blotting, what can you say about the initial ELISA result?
The initial ELISA result was a false positive but the Western blot confirmed this positive as false.
20.Describe the role of the developing buffer in the Western blotting technique. The developing buffer contains the secondaryantibody that the enzyme
alkaline phosphatase are conjugated to.
21.Which patient(s) tested positive for HIV using the Western blotting technique? Patient C tested positive for HIV
22.Using the choices listed below, decide which of the criteria apply to the indirect ELISA, which apply to the Western blotting technique, and which
apply to both. Use "E" if it applies to ELISA, "W" for Western blotting, and "B" for both.

Criteria:
E1. It tests for the presence of antibodies in the patient sample.
W2. Electrophoresis is used.
E3. Substrates that change from colorless to colored are used.
B4. An enzyme–linked secondary antibody is used.
E5. Antibodies against individual antigens are detected.
E6. The reaction takes place in a well in a microtiter plate.
E7. An antigen–antibody reaction is detected.
E8. An optical density is read to determine whether or not a sample is

Fluorescence Spectroscopy And The Emission Wavelength Of...
Abstract Fluorescent properties are used to study protein and small molecule interactions. Fluorescent spectrophotometry determined the excitation
wavelength of eosin isothiocyanate to be 525 nm and its emission wavelength was 545 nm. Glycogen phosphorylase was similarly studied. The
excitation wavelength was 330 nm and the emission wavelength was 360 nm. The emission wavelength could be indicative of the presence of the
fluorophore, tryptophan. The potential interaction between bovine serum albumin (BSA) and 1–anilino–8–naphthalene sulfonic acid (ANS) was
studied through fluorescence and fluorescence resonance energy transfer (FRET). BSA had an emission wavelength of 358 nm which could also be
indicative of a tryptophan. When BSA and ANS were mixed together, the emission wavelength was longer suggesting that the molecules interact with
each other and follow FRET.
Introduction
Fluorescence spectroscopy is a useful technique to determine the proteins and nucleic acids of a macromolecule. Some molecules, fluorophores, can
absorb light at certain wavelengths and then emit it at another wavelength. When the molecule absorbs light it is excitation and the light released is
emission. The light particles, photons, hit a fluorophore to excite the molecule (Voet et. al 2013). Excitation occurs by the valence electrons of the
molecule receiving energy and moving to a higher energy state from the ground state. The excited electrons will later return to the ground state because
they

Advantages And Disadvantages Of Dihydropyrimidin-2
Recently, the synthetic chemists have gradually taken concern of the compounds containing nucleus of 3,4–dihydropyrimidin–2(1H)–one, which have
a series of pharmacological characteristics, such as calcium channels blockers, antioxidant, anticancer, and antiinп¬‚ammatory activities [1–5]. But their
efficiently utilizing is boxed in low water solubility. The sulfonate group is strongly acidic. As a result, it is expected that the solubility of
3,4–dihydropyrimidin–2(1H)–one derivatives could be greatly improved by introducing the sulfonte group. In this essay, through sulphonation with
good hydrosoluble prosperities, we have successfully obtained two creative 3,4–dihydropyrimidin–2(1H)–one derivatives 1, 2 (structures shown in
Scheme 1).
Serum albumin (SA) is one of the most ... Show more content on Helpwriting.net ...
The reaction scheme is showed in Scheme 1. Brieп¬‚y, using sulfamic acid as a catalyst, DHPMs1 and DHPMs2 were synthesized by ethanol
thermal method. Urea (0.90 g, 15 mmol), acetaldehyde (0.44 g, 10 mmol) / benzaldehyde (1.06 g, 10 mmol), ethyl acetoacetate (1.56 g, 12 mmol)
and sulfamic acid (0.8 g, 8 mmol) were dissolved in 10 mL ethanol to form a clear solution, which was settled in a 25 mL teflon–sealed autoclave and
maintaining at 80 ВєC for 2 h. After cooling, the reaction mixture was cleaned with cold water and 50 % ethanol, and the residue recrystallized from
95 % ethanol to afford the pure product. DHPMs1: Slightly water–soluble white powder; Yield = 1.31 g (71 %); Mp 192 – 193; formula: C9H14N2O3;
FW: 198.22; IR (KBr) cm–1пјљ3244, 3118, 2978, 1702, 1654пј›ESI–MS (m/z): 197.33 (M + H). DHPMs2: Slightly water–soluble white powder;
Yield = 2.43 g (93.6 %); Mp 206 – 207; formula: C14H16N2O3; FW: 260.29; IR (KBr) cm–1пјљ3235, 3108, 2975, 1702, 1645пј›ESI–MS (m/z):
259.18 (M +

Succinate Dehydrogenase Lab
Introduction
In this Succinate Dehydrogenase (SDH) project, we were expected to learn the process of cell fractionation, the importance of the Bradford assay, how
enzymes work what factors affect them, how to adequately micropipette, how to generate a proper hypothesis and test it, and finally, interpret our
results and write a conclusion which answers the central question of this lab: how will the assigned reagent affect SDH activity?
The SDH project was completed over a span of three weeks, where we had a primary concentration each week that revolved around a core principal
of this project. In week two, which was the first week of this project, we blended and centrifuged our cell fractions in order to isolate them, and then
we used a sample ... Show more content on Helpwriting.net ...
absorbance. We graphed the results from the Bradford Assay on excel and created a linear standard curve and equation for it, which was used to
determine the protein concentration in our cellular fractions. The data from the cellular fractions was used to discover the amount of specific SDH
activity in lysates. Furthermore, we also conducted the SDH assay in order to gain proper and knowledgeable experience about how to assess SDH
activity for our crude and mitochondrial cellular fractions, and we also worked with enzymes to further learn about how they work and how they are
affected through various factors. The second week of this lab was rather heavy; it had many components to it, each equally important for our end result
and conclusion. The concentration in week two was on the Bradford assay, which was imperative since it measured protein concentration, and
practicing with the SDH assay, which measured the activity of our crude and mitochondrial fractions, and was therefore vital in the final week of the
project.
During the final week of the project, we performed the SDH assay twice to test our hypothesis and answer the primary question of this project: the
impact of the assigned reagent (Malonate for us) on SDH activity in our mitochondrial fraction. Week three's concentration was on correctly performing
the SDH assay twice in order to minimize human error, and to interpret our data for our lab

Total Serum Protein Determination Lab Report
Introduction
Total serum protein determination is a standard clinical test that is often used for the diagnosis of diseases involving the kidneys, liver or bone marrow
and other metabolic or nutritional disorders1. Blood plasma contains approximately 7% proteins by weight and the fluid that remains after the clotting
factors are removed from the plasma is the serum2. Albumin and globulin are the most common blood proteins, and the total protein is the combined
measure of the two3. The most abundant protein is albumin, which acts as a carrier of many different molecules in the blood and helps maintain blood
volume2. The globulin proteins include many enzymes and elements of the immune system such as antibodies and immunoglobulins2. Changes in ...
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This method is a chromogenic test that uses biuret reagent to detect the presence of peptide bonds in blood serum12. The active ingredients in the
reagent are sodium hydroxide, potassium sodium tartrate and copper sulphate pentahydrate13. The development of a violet colour after incubation at
room temperature is indicative of the presence of proteins and peptides in the blood serum12. A coloured coordination complex forms when the copper
ions (Cu2+) binds to the deprotonated nitrogen atoms that form the peptide bonds of the amino acids12. Reactions can also occur between Cu2+ and
the nitrogen in the amide groups and the imidazole rings12. The colour intensifies after a period of incubation as the alkalinity of the solution increases
and more deprotonated nitrogen atoms become available12. The concentration of the protein and the colour intensity are proportional, thereby
exhibiting a linear relationship in accordance with the Beer–Lambert law14. Absorbance is measured by spectrophotometer, О»max = 540

Subject Identification And Enrollment For Alzheimer 's...
Subject identification and enrollment Subjects recruited for this study consisted of individuals aged 60 to 85, with 10 or more years of education
or more. Those diagnosed with Alzheimer's disease (AD) and mild cognitive impairment (MCI) were recruited to the study at the Memory Clinic
of the Jewish General Hospital (JGH) in MontrГ©al, and normal community elderly volunteers were recruited through advertising. Subjects
received standardized clinical, mental status, neuropsychological, and cognitive evaluation. Depression was excluded via the Yesavage Geriatric
Depression Scale (Geriatric Depression scale > 15) [37]. None had evidence of cerebral infarct on CT, and all subjects scored less than 4 on the
Hachinski ischemic scale [38]. No individuals had evidence of significant heart disease, alcoholism, or drug use and all showed normal routine blood
work. Subjects were classified as normal if they lacked subjective memory complaints, and scored at or above 26 on the Montreal Cognitive
Assessment test [39], and scored above 25 on the Folstein Mini–mental state exam (MMSE) [40]. Their average MoCA score was 27 (range 25–29).
The AD patients received the diagnosis of mild, moderate, or severe probable AD according to standard clinical criteria [41]. They had mild,
moderate, or severe probable AD [42], scored below 26 on the Montreal Cognitive Assessment test [39], scored between 4 and 25 on the Folstein
Mini–mental state exam (MMSE) [40], and showed

The Specificity of Albumin Binding Experiment
The specificity of albumin binding experiment was to determine the binding interactions that occur between serum albumin and three synthetic dyes
with the use of electrophoretic procedure. Whole blood, or plasma. Clots upon standing and if the clot is removed, the remaining straw colored fluid is
called serum. The major protein in serum is albumin which functions as a carrier molecule for the transport of certain small molecular weight
compounds in blood. Molecules that bind to serum albumin are fatty acids, hormones and some synthetic dyes. In this experiment the synthetic dyes
used are Bromophenol Blue, Ponceau S and Orange G. we observed that free dyes not bound to albumin migrate faster that albumin or dyes bound to
albumin. This... Show more content on Helpwriting.net ...
The 2nd half for the gel is our experiment compared to Anderson group 6. The 1st layer in all 3 pictures is the bound dye, the 2nd layer is the free dye.
Fig1 shows on the right group one next to it, 1st half of our result on electrophroresis gel. The 2nd half for the gel is our experiment compared to
Anderson group 6. The 1st layer in all 3 pictures is the bound dye, the 2nd layer is the free dye.
Group 1, the gel picture and Andersons experiment are closely related. The binding did occur, in the 1st lane of group 1the dye was free therefore no
binding occurred because no Bovine Serum Albumin (BSA) was present in the mixture. While in lanes 2 to 4 binding did occur because there was BSA
in the mixture, with the higest binding occurring in lane 4, this is because there was no buffer to interrupt the binding. (saturation).
Group 6, which is the 2nd half of the gel picture, does not correspond with Anderson results, no visible binding occurred. This group had two different
dyes and SDS included in the mixture also. In Andersons version although two different dyes were used, it didn't affect his results, it's the SDS that

How To Write An Enzyme Lab Report
Enzymes are a diverse category of proteins that are used in various biochemical systems to catalyze a particular reaction. They are vital in increasing
the rate of reactions by up to 1017 which aids in the functioning of cells while having implications in medicine, biotechnology, and the food
/pharmaceutical industries.1 The purpose of this experiment was to extract and purify an enzyme of interest known as acid phosphatase. The process of
enzyme isolation is essential for using these proteins outside of a cell, unlocking their benefits for practical uses. However, the procedure involves
various steps with inherent risk of denaturing the enzyme via common biochemical such as inhibitors and oxidizing/proteolytic agents, thus the
effectiveness of the enzyme is analyzed by taking samples (aliquots) at each purification step (fraction).1 The enzyme of interest in this experiment was
initially extracted from wheat germ via osmotic rupturing of the cellular membrane to release the water–soluble protein.1 This solution was then
centrifuged to sediment various cellular debris into a pellet, leaving a solution of various hydrophilic solutes, including acid phosphates. ... Show more
This procedure involves exposing the enzyme to two concentrations of the salt (35% and 57%) in two different steps. This is called salt fractionation
and effectively separates large groups of proteins, lipids, carbohydrates, and DNA. The process works by causing the highly hydrophobic molecules to
become insoluble at 35% and the partially hydrophobic (acid phosphatase) to precipitate at 57%, leaving the most hydrophilic solutes in solution.1 The
salt concentration and type may be varied based on the isolation compound of interest, for the overlying principle of this technique, selective
dehydration, remains the

Sam Case Study
Viperitoxin decreased the twitch response by around 52–53% and decreased the KCl response by around 113%. There was no CCh response as it was
not tested. Taipoxin decreased the twitch response by 83%, whilst the CCh response did not change, but increased the twitch response of KCl by 18%.
For О±–bungarotoxin, the twitch response decreased by 97%, whereas the CCh response by decreased 84% and the KCl response decreased by 19%.
Discussion of results 1.What is the relevance of the "BSA" data and why is it important to include these data when making conclusions about the
mechanisms of action of the 3 toxins? A: The Bovine Albumin Serum 'BSA' is a protein derived from cows and is commonly utilised in ELISA tests
to prevent non–specific binding from occurring. Conversely, it also serves as a protein supplement. The BSA is... Show more content on Helpwriting.net
...
Based on your conclusions, which, if any, of the venoms would likely cause a paralysis similar to what Sam experienced when he was bitten by the
snake. Explain your reasoning. This study does not conclude which venom is responsible for Sam's paralysis, when bitten; Sam experienced symptoms
such as flaccid paralysis. Any of the three venoms tested (Viperitoxin, A–bungarotoxin and Taipoxin) could cause similar levels of paralysis since all
three toxins affect muscle contraction; such as the post–synapse, pre–synapse or directly on the muscle. It is difficult to pin–point the specific
neurotoxin due to the similarity of causation of the symptom of flaccid paralysis. 4.What information does this study provide about the reversibility of
each of the toxins? Explain your response. This study does not provide adequate information on the reversibility of the three toxins presented. There is
no data for each of the toxins on the twitch responses–post washing the toxin out of the organ bath. Omitting this data inhibits the ability for this study
to reach a conclusion, in terms of finding out on whether there are reversibility qualities of each

Separation Of The Proteins Involved
Separation of the proteins involved in both species blood and serum revealed that each was composed of a major protein. A prominent protein in the
rabbit blood was found accumulating at approximately 13kDa, as shown in Figure 1 and Figure 2. It was concluded that this protein was most likely
hemoglobin, given the weight and large amount present in all blood due to its necessary function of transporting oxygen and carbon dioxide throughout
the body. Hemoglobin is a "tetrameric protein of about 64.5 kDa, consisting of two –chains and two –chains" which breaks down into each subunit
having an approximate weight of 15kDa (Yu et al, 1997). This was in correlation with the data present in Figure 1, given that the major protein present
in rabbit blood was approximately 13kDa, which would be the subunits that make up hemoglobin.
We hypothesized that similar proteins would also be present in bovine blood, yet the major band present in this sample was located at much higher
molecular weight of approximately 45 kDa. Potential causes in weight differences could be the longer amino acid sequence present in rabbit
hemoglobin, 170 amino acids, versus the 145 amino acids present in bovine hemoglobin, yet this would result in bovine being further down on the gel
versus what is actually shown (Altschul et al, 1997). Given the data obtained, potential misidentification of hemoglobin appearing as the main protein
present or possible error in lane one (rabbit blood), could explain the anomaly in

Reaction Of The Maillard Reaction
Introduction
The non–enzymatic reaction of reducing sugars with amino groups of amino acids, peptides and proteins is called as non–enzymatic glycosylation or
simply glycation. This non–enzymatic reaction ultimately results in the formation of complex brown pigments and protein–protein crosslinks that were
first studied under defined conditions by Maillard in the early 1900s (Maillard and Gautier, 1912) and the reaction came to known as the Maillard
reaction. There is a condensation reaction between reducing sugar and free amino acids at the amino terminus or susceptible Оµ–amino group of lysine
residues of proteins producing labile N–substituted glycosylamine (Schiff base) that may undergo two sequential rearrangements, forming a stable
Amadori product (Fayle & Gerrard, 2002). The reducing sugars namely, glucose, fructose, galactose, mannose, ribose and certain carbohydrate
relatives are inherently reactive toward nucleophilic groups. Glucose is the least reactive among common sugars because it exists in least concentration
in the open chain form (Paulsen and Pflughaupt, 1980). Amadori rearrangement was a key early step in the Maillard reaction (Hodge, 1955). Early
glycation directs the formation of Schiff's bases and Amadori products that further oxidized to advanced glycated end products (AGEs) (Chetyrkin et
al., 2008). Studies have shown that the glycation process (in the early stage where the interaction of sugars, such as glucose, with free amino groups of
lysine and

The Criteria For American College Of Rheumatology ( Acr )...
Subjects
Thirty patients that fulfilled the criteria of American College of Rheumatology (ACR) for RA will be included in our study. According to 2010 ACR
Classification Criteria for RA equal or more than 6 criteria = definite RA
Thirteen healthy persons were also, included in our study as a standard group.
All persons were subjected to:
Full history taking include:
Age and sex.
Anti–Rheumatic drugs administration.
Underlying medical conditions: liver diseases, renal failure, malignant diseases, infectious diseases or other inflammatory diseases.
Exclusion criteria:
1.Renal or hepatic disorders
2.Platelets < 150000 & WBCs < 3500
3.Other autoimmune disease
4.Malignancy
5.Cardiac diseases
Clinical examination:
All patients will be assessed according to 2010 ACR Classification Criteria for RA equal or more than 6 criteria = definite RA.
Disease activity was assessed with the 28–joint disease activity score (DAS28).
Routine Laboratory investigation:

Complete blood picture was done for healthy and RA patient groups.
The erythrocyte sedimentation rate (ESR) and C–reactive protein (CRP) will be done.
Rheumatoid factor (RF)–IgM was determined in all patients by means of latex agglutination.
Serum levels of anti–cyclic citrullinated peptide (anti–CCP) were measured by ELISA.
Concerning the administered basic therapy, all patients received MTX dose 17.5 mg/week. In addition, patients received hydroxychloroquine

Mandatory Synthesis Essay
Beef Allergies and Nutritional Alternatives on Those Affected There have been a significant number of studies indicating a growing beef allergic
response in our population (Fiocchi et al. 2000). Moreover, a high incidence of atopic dermatitis among children and the elderly raises concerns
because of its direct impact with nutrition and well–being (Fiocchi et al. 2000). As a result, different types of food challenges, test on the skin such as
skin–prick test(SPT) and careful observation of immediate whale–and–flare reactions have emerged (Fiocchi et al. 2000). Diagnosing Bovine Serum
Albumin(BSA), Ovine Serum Albumin(OSA) and Alpha–gal as the main anaphylaxis agents that Immunoglobulin G (IgG) response is directed.
Resulting in different nutritional... Show more content on Helpwriting.net ...
Anaphylaxis responses are becoming more common due to genetic and environmental factors (Saleh et al. 2012). BSA, OSA and Alpha–gal continue
to be the main allergic agents known today (McNeill and Van–Elswyk 2012). Health and well–being g hand–to–hand with a proper diet. As a result of
atopic dermatitis, different options have been adopted for proper intake of micronutrients and essential vitamins of those affected (Asp et al. 2012). In
specific, the children and elderly still remain the most affected due to this condition (Saleh et al. 2012). Asp, M.L., Richardson, J.R., Collene, A.L.,
Droll, K.R. and Belury, M.A. Dietary protein and beef consumption predict for markers of muscles mass and nutrition status in older adults.
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Mol. Allergy. 10(5), 1–11(2012).
Van–Nunen, S. Tick–induced allergies: mammalian meat allergy, tick anaphylaxis and their significance. Asia.Pac.Allergy. 5(1),

Enzyme Linked Immunisirbent Assay
The major use of Enzyme Linked Immunosorbent Assay (ELISA) is to titrate antibodies against specific antigens (Crowther, 1995). This method
involves an enzyme, which reacts with a colourless substrate to form a coloured product, is covalently linked to a specific antibody that recognises a
specific antigen. If the antigen is present, the antibody–enzyme complex will bind to it, and the enzyme component of the antibody–enzyme complex
will catalyse the reaction which results in a coloured product (Berg et al, 2002). The presence of the coloured product indicates that an antigen is
present. The ELISA, which is rapid and convenient, can detect less than a nanogram (10–9 g) of a protein and can be performed with either polyclonal
antibodies, which are antibodies that can recognise multiple epitopes on any one antigen or monoclonal antibodies, which are antibodies that can only
detect one epitope on the antigen (Guo et al, 2006). However, the use of monoclonal antibodies gives more reliable and accurate results due to their
specificity as monoclonal antibodies are very effective at acting as the primary antibody in an assay, or for detecting antigens within a tissue. Also, they
will usually result in reduced background staining compared to that of polyclonal antibodies.
Indirect ELISA and direct ELISA Different types of ELISA's have been developed with modifications to the basic steps. The key step with the ELISA
assay is the direct or indirect detection of antigen by

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