Wnt Signaling Pathway

Wnt Signaling Pathway
The notch pathway is a new avenue in battling melanoma which holds promise in preventing
chemoresistance. The notch pathway is an evolutionarily conserved embryonic cell pathway
important for cell fate and differentiation.32 Studies have suggested that dysregulated notch
signaling can prolong and confer life to cancer stem cells, which are thought to be important in
tumor chemoresistance.33 Other studies have shown that notch activation promotes melanoma
tumorigenesis and progression.32 Thus, therapeutic benefit of targeting notch signaling includes
preventing tumor angiogenesis, cancer stem cell depletion, and cell death.34 The notch receptor
family consists of four transmembrane receptors (Notch 1–4).35 Recent research on melanoma cells
... Show more content on Helpwriting.net ...
The mechanisms underlying melanoma invasion are presently poorly understood. The Hippo
pathway (the Salvador–Warts–Hippo) is an evolutionarily conserved mechanism in charge of tissue
and cell growth.42 Downstream in the Hippo pathway are YAP and TAZ effector proteins which are
amplified in many cancers and promote epithelial–mesenchymal transition–a frequent hallmark of
metastasis.43 In a model of skin reconstruct, YAP overexpression led to increased melanoma cell
invasiveness, and YAP knockdown led to decreased metastasis potential.43 Thus, mutations in the
Hippo pathway yield increased activation of YAP and TAZ, which promote metastasis regardless of
BRAF mutation status in melanomas.42,44 Current research is examining verteprofin,44 a molecule
that inhibits YAP function and whether or not it can successfully regulate Hippo effector functions
and subsequently melanoma
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Mouse Model And Sample Size
Mouse model and Sample Size
This study will be using 8 week old Smn1tm1Hung Tg(SMN2)2Hung mice obtained from the
Jackson Laboratory. This specific strain of mice will be selected as they are double mutants; they
will have undergone an exon 7 deletion in their SMN1 gene resulting in the development of SMA,
and they will have a human SMN2 transgene, since mice do not normally have SMN2 genes. The
mice will be homozygous for the SMN1 with the knockout, this is important because over 95% of
SMA cases are due to the homozygous deletion of SMN1 genes. They will also be homozygous for
the SMN2 transgene as SMN2 hemizygous mice do not survive past 16 days postnatal [The Jackson
Laboratory, 2017].
Using these mice, we will carry out in vitro ... Show more content on Helpwriting.net ...
The power required for this experiment will be 80% to minimise the number of experimental mice
used, since the difference we hope to measure is significantly large. A significance value of 0.05 will
be used to limit the number of times type 1 errors may occur. With an effect size of 0.8, a power
value of 0.8, and a significance of 0.05, a sample size of 52 is calculated, with 26 mice embryos in
the control group and 26 in the treated group [AICBT Ltd, 2017]. Designing sgRNAs Single–guided
RNAs (sgRNAs) are required to guide the Cas9 endonucleases [Long, et al., 2014]. We need to
design suitable sgRNAs to use in our experiment and test the feasibility of CRISPR–Cas9–mediated
genetic repair at the cellular level before we can inject the system into mice zygotes. Around five
different sgRNAs can be designed, targeting different regions of the mutant SMN1 gene – three
targeting regions of the base pair deletion, one targeting the region immediately downstream of the
base pair deletion and one targeting the normal sequence upstream of the deletion [Wu, et al., 2013].
The specificity of each of these sgRNAs can be tested through transfecting plasmids with the Cas9
and each of the different sgRNAs and place into wild–type embryonic stem cells. PCR can be
carried out for each sgRNA and their products, that we can assume to be the target region, can be
amplified from the subsequent embryonic stem

The Mechanisms Linking Ribosomal Translation And Pre Mrna...
Investigation of the mechanisms linking ribosomal translation and pre–mRNA processing in yeast
Nine–Month Report submitted by
Marija Petrić
ID: 1454106
Supervisor:
Dr Saverio Brogna
Contents
Contents
1. Introduction 3
1.1. Eukaryotic gene expression 3
1.2. Quality control of eukaryotic gene expression 5
1.3. Nonsense mediated mRNA decay 6
1.4. Factors involved in NMD 7
1.5. NMD models 17
1.6. Ribosome release model 21
1.7. Role of UPF1 in the nucleus 23
2. Objectives 25
3. Materials and methods 27
3.1. Schizosaccharomyces pombe strains 27
3.2. S. pombe media 27
3.3. S. pombe culturing and maintenance 27
3.4. PCR based gene targeting – one step PCR approach 28
3.5. PCR based gene targeting – two step PCR approach 30
3.5. Agarose gel electrophoresis and gel extraction 32
3.6. Transformation of S. pombe strains for PCR based gene targeting 32
3.7. Colony PCR 33
3.8. pDUAL expression vector 34
3.9. Polysome profile 36
3.9.1. Preparation of sucrose gradients 36
3.9.2. Cell lysis 36
3.9.3. Sedimentation of translation complexes 37
3.9.4. RNA and protein precipitation from polysomal fractions 38

3.10. SDS–PAGE and Western blot 39
4. Results 41
4.1. How does UPF1 bind its substrates? 41
4.1.1. Introduction 41
4.1.2. Results and discussion 43
4.2. What is the mechanism of NMD? 48
4.2.1. Optimisation of polysome profile analysis 48
4.2.2. Ribosome release model 49
1. Introduction
1.1. Eukaryotic gene expression Gene expression is the basic cellular process which converts

Yy1 Research Paper
Yin Yang 1 in HCC: YY1 is a transcription factor with complex biological functions, including
apoptosis, tumorigenesis, development and differentiationYY1 is a member of the Polycomb Group
protein family, a group of homeobox gene receptors that play critical roles in hematopoiesis and cell
cycle control [26, 27]. The human YY1 gene is located on the telomere region of human
chromosome 14 at the segment q32.2. It consists of five highly conserved exons encoding a protein
of 14 amino acids in length, and an estimated molecular weight of 44 kDa. The YY1 protein
contains four C2H2–type zinc–finger motifs with two specific domains that characterize its function
as an activator or repressor. YY1 is a phosphoprotein with a half–life of 3.5 hours. ... Show more
content on Helpwriting.net ...
A large body of literature has documented hyperactivated AKT signaling in human solid tumors and
hematological malignancies[34]. The PI3K/AKT/mTOR signaling pathway plays an important role
in HCC and is activated in 30%–50% of HCC[35]. Sorafenib does not target this pathway.
AKT/mTOR is critical for regulating growth and proliferation in any cell type. Data from HCC
clinical trials have revealed that AKT/mTOR pathway is upregulated upon treatment and therefore
the cancer acquires resistance to therapy. Cyclin D1 is a downstream target of AKT, which directly
regulates cell cycle progression. Of the three highly conserved AKT isoforms, AKT1 is involved in
regulating cell proliferation[36]. Accumulating research suggests that PI3K/AKT activation after
sorafenib treatment plays a pivotal role in tumor progression in HCC. p–AKT levels increase in
HCC cell lines after treating them with sorafenib [11, 37]. Targeting AKT1 will also suppress the
activity of its downstream effector pathway

James Rothman, Randy Schekman and Thomas Südhof's...
The 2013 Nobel Prize in Physiology and Medicine was awarded to James Rothman, Randy
Schekman and Thomas Südhof for the work that they did on transport vesicles within the cellular
membrane. The recipients discovered how the cellular transport system was organized so that
transport material was delivered to the correct site with proper timing. Rothman discovered how the
vesicle is able to fuse with a cell membrane or organelle to deliver its contents. Schekman through
the study of yeast isolated the genes required to code for vesicle transport. Südhof found the signals
that tell vesicles when to release their contents.
Schekman studied the cellular transport of system of yeast and documented his discoveries in his
1990 paper Distinct ... Show more content on Helpwriting.net ...
Further research showed that the NSF protein is interchangeable with the SEC18 gene (previously
discovered by Schekman) for vesicle binding in mammals and yeast. Rothman also discovered that
a calcium shift must occur before The NSF and SNAP proteins bind, so that the correct
conformation on the membrane is available. By studying the interaction of between NSF and SNAP
protein complexes and target cell membranes a derivative of the SNAP proteins were found, called
SNARE complexes. The SNARE complexes were needed for greater docking ability in the transport
vesicles.
Südhof discovered in his 1990 paper Phospholipid binding by a synaptic vesicle protein
homologous to the regulatory region of protein kinase C that vesicle binding is a specific and
precise process that is regulated by neurotransmitter release. The release of vesicles for membrane
bidding is monitored by the influxes of Ca2+ ions into the cell. An increase of Ca2+ triggers the
vesicle to bind to the phospholipid bilayer of a cell. Once bound the Ca2+ triggers a
neurotransmitter that signals the bound vesicle to release its contents into the cell membrane by
exocytosis. Südhof also confirmed that in order for certain vesicle to bind to target membranes a
SNARE protein complex must be present in order to promote vesicle binding.
When Schekman discovered the seven specific genes required to code and assist in transport, it

Marketing Vs Showcasing Essay
AMITY COLLEGE OF COMMERCE AND FINANCE
TERM PAPER EVALUATION
"TRADITIONAL MARKETING vs. E–MARKETING: which is more effective and efficient?"
Submitted to– Ms. Sarika Submitted by– Nancy Rahuja Bcom. (honrs.) Section–B A3104613126
CHAPTER–1
INTRODUCTION
Marketing has practically been around always in one structure or an alternate. Since the day when
people initially began exchanging whatever it was that they initially exchanged, promoting was
there. Showcasing was the stories they used to persuade different people to exchange. People have
progressed beyond anyone 's expectations from that point forward, (Well, we like to think we have)
and showcasing has as well.
The systems for showcasing have changed and enhanced, and we 've turned into a considerable
measure more effective at telling our stories and getting our advertising messages out there.
emarketing is the result of the gathering between present day correspondence

4. 1 Functional Classifications Of Functional Functions
4.1 Functional classification of dysregulated proteins
It was shown that MPS VII is characterized by accumulation of ubiquitin and neurofilament, but not
PHF–τ, α–synuclein or apoptosis [20]. Similarly, significant alterations in the ubiquitin–proteasome
system (UPS), but not proteins associated with apoptosis or PHF–τ and α–synuclein, were identified
in this study. Previous transcriptomic and proteomic analyses of MPS VII mouse brain identified
many dysregulated proteins involved in metabolism and cytoskeleton [21,22]. Additionally, these
studies found that neuroinflammation is also upregulated, but not to a large extent. Similar results
were observed in this study, indicating the reliability of the proteomics approach. We also examined
... Show more content on Helpwriting.net ...
Dysregulation of cytoskeletal genes has also been seen in MPS I mouse brains [4], MPS IIIB mouse
brains [26] and MPS VII dog hearts [27]. Additionally, low density lipoprotein–related protein 1B
(LRP1B), dynamin–1 (DMN1) and dynamin–related protein 1 (DNM1L), which are involved in the
process of endocytosis, are downregulated. Vesicle associated membrane protein 2 (VAMP2) is
involved in the targeting and/or fusion of transport vesicles to their target membrane [28]. These
results further suggest impaired lysosomal membrane trafficking pathways in MPS VII.
Based on previous reports and results in this study, we hypothesize that GAG accumulation leads to
alterations in the cytoskeletal system and thereby impairs lysosomal membrane pathways including
endocytosis, exocytosis, autophagy and vesicle transport. Notably, endocytosis defects in brain
endothelial cells may contribute to low transcytosis, making enzyme replacement therapy more
difficult to achieve neurological benefits. Therefore, targeting specific pathways involved in
transcytosis may be an effective strategy to enhance enzyme delivery to the brain in lysosomal
diseases including MPS VII.
4.1.2 Proteostasis
Many neurodegenerative diseases share a common molecular signature: disruption of proteostasis
[29]. This often manifests as an accumulation of ubiquitylated proteins, with evidence for a robust
contribution to

Why Mitochondria And The Cell With Versatile Operating...
Abstract:
Although mitochondria and chloroplasts encodes handful of their own proteins, however, majority
of their required proteins have to be synthesized in the cytosol and translocated into their correct
destinations using specialized transporting networks: TOM/TIM complexes in mitochondria and
TOC/TIC complexes in chloroplasts. Molecular chaperones play critical roles in facilitating
functional competent–protein import from the cytosol to their correct destinations, utilizing catalytic
motor components and other transporting channels. Biogenesis of both mitochondria and
chloroplasts and their maintenances in terms of transcription, translation and protein import into
their various numerous compartments requires very tight coordination to overcome energetic
barriers, protein aggregation and protein degradation. This essay describes molecular chaperones
involvement in protein import from the cytosol into these double membraned organelle
compartments.
Introduction:
Mitochondria are the powerhouse of the cell with versatile operating systems (i.e. converts energy
derived from foods into cellular energy e.g. ATP, Amino Acid and lipid metabolisms, iron–sulphur
clusters and haem biosynthesis, and also the regulation of apoptosis) (Harbauer et al., 2014,
Bolender et al., 2008, Wiedemann et al., 2004). Equally, chloroplasts are also very versatile and
operates several metabolic and cellular processes (i.e. photosynthesis, amino acid and lipid
metabolism, cellular signalling and

Biodegradable Nanoparticles for Drug Delivery
Review: Biodegradable Nanoparticles for Drug Delivery
Introduction
With the many innovations in the disciplines of nanotechnology and drug delivery their merger is a
very advantages strategy transport of small molecular weight drugs and macromolecules for
localized or targeted delivery to the tissue of interest [1]. Nanotechnology will provided an outlet for
conveying therapeutic agents in biocompatible nanocomposites such as nanoparticles, nanocapsules,
micellar systems, and conjugates. With the rewards of the nano scale and interactions of these
delivery systems, remedies can be used to provide targeted delivery of drugs, to improve oral
bioavailability, to sustain drug/gene effect in target tissue, to solubilize drugs for intravascular
delivery, and to improve the stability of therapeutic agents against enzymatic degradation, especially
of protein, peptide, and nucleic acids drugs [1]. By tailoring polymer qualities, on can specify
design factors that can control the release of therapeutic agents to provide excellent targeting for
optimal efficacy. The ability to theses distant destinations, nanoparticles are led by bio specific
ligand which could direct them to the target tissue. The prime emphasis on the review will be the
mechanism of their intracellular uptake, different pathways of their uptake, intracellular trafficking
and sorting into different intracellular compartments, and the mechanism of enhanced therapeutic
efficacy of the nanoparticle–encapsulated

Dynamic Research Design
In designing an experiment, one must be able to control as many variables as possible so the results
can be interpreted as significant in knowing that no outside errors could occur. This basic
component of experimental design allows for one to manipulate variables to achieve the desired
outcome. A good scientist keeps this inquisitiveness in order to advance themselves in the project
they are working on and the next experiment. The dynamic that exists between a researcher and
their project is lead by intuition and persistence. This constant effort is made to get an experiment
to, hopefully, demonstrate some phenomena. I have exercised all of the above qualities in many
different research experiences. Most notably, the research experience I ... Show more content on
Helpwriting.net ...
Such a deficit of protein can disturb numerous mitochondrial pathways, such as membrane potential
and respiratory function. Some of the most essential functions include the tricarboxylic acid cycle,
oxidative phosphorylation, defense against reactive oxygen species, and mitochondrial DNA repair.
The most significant decrease in abundance are Succinate Dehydrogenase Complex, Subunit A
(SDHA) and Tu Translation Elongation Factor (TUFM). Interestingly, SDHA couples the
tricarboxylic acid cycle and oxidative phosphorylation pathways. TUFM participates in protein
translation. These deficits could interfere with mitochondrial ATP production and may increased
oxidative damage of mitochondrial proteins and DNA due to a lack of machinery to correct it. The
deficit of the mitochondrial proteome strongly suggests a role in the neurodegeneration in

Applying Autophagy and Apoptosis for Strategies to improve...
The pathways controlling autophagy and apoptosis should be targeted as the strategies to improve
treatment of glioblastoma. The proteins to be targeted are Beclin, p62 and EGFR.
Survival related or anti–apoptotic protein such as Bcl2 were found to be upregulated in
glioblastomas whereas autophagy proteins like Beclin were found to be downregulated. The
autohagy (Atg) proteins function at several discrete but continuous steps in autophagy which
include induction or selection/packaging of cargo, vesicle nucleation, vesicle elongation, vesicle
docking and fusion with lysosomes, and degradation of vesicular contents. There is significant
evidence showing overlap between these two pathways. Hence, we choose to target these pathways
and ... Show more content on Helpwriting.net ...
p62 is an autophagy selective substrate and it accumulates when autophagy activity is reduced.
Indeed, the level of p62 is often used as an indicator of autophagy activity. Interestingly, p62 levels
are commonly upregulated in human tumors and genetic ablation of p62 in various tumor models
has been shown to reduce the tumorigenesis occurring because of autophagy‐deficiency.
EGFR: The most common genetic mutations / alterations seen in gliomas are the amplification of
EGFR, expression of EGFR VIII mutant and homozygous / hemizygous deletion of PTEN and NF–
1. The aberrant signals that result from these mutations interact with the PI3KAkt– mTOR pathway
and are responsible for promoting survival and chemoresistance in gliomas. Therefore targeting the
receptor tyrosine kinases (RTKs) with either monoclonal antibodies or small molecule inhibitors
emerged as a promising therapeutic strategy . Clinical studies with small molecule inhibitors of
EGFR, such as erlotinib and gefitinib, have been disappointing in gliomas. Similarly, monoclonal
antibodies against EGFR, cetuximab and panitumumab, have only a cytostatic effect in glioma cell
lines. The abundance of multiple types of RTKs along with the frequent deletion of PTEN in
gliomas may account for the lack of effectiveness of single agent tyrosine kinase inhibitors.
In

Tumor Busting Virus Essay
Reading an article and learning about "tumor–busting viruses" is very compelling, to say the least.
Since it seems almost everyone has endured the pain of cancer or has a loved one who has been
diagnosed with some form of cancer, it is refreshing to see that researchers and scientists are coming
together to find fresh techniques to help heal this tragic illness.
This new remedy that scientists are experimenting with is called virotherapy. Virotherapy was
defined in the article as viruses that have been genetically engineered to find and kill cancer cells
while leaving healthy cells untouched. A treatment such as this has been a favorable one considering
how the traditional cancer chemotherapies have been known to not only attack cancer ... Show more
content on Helpwriting.net ...
Because all people, roughly, have been exposed to adenovirus, humans hold antibodies from the
immune system to attack and kill the virus. Therefore, using adenoviruses in cancer virotherapy may
result in some symptoms that seem like indicators of the flu. This is because the immune system has
been trained to respond to the virus by attacking it and eliminating it from the body, which will also
stamp out the therapy. Researchers have been attempting to fight the side effects by giving
immunosuppressive drugs while the patient is on the virotherapy as well as altering the adenoviruses
so that the immune system will not respond to the virus. To create the best version of virotherapy,
scientists are discovering two ways to be confident the viruses hit target cells with no other harm to
other cells. One strategy virotherapy researchers are exploring is called transductional targeting.
With this form of targeting cancer cells, researchers are modifying the viruses used so they are only
attracted to only cancerous cells to contaminate them. To prevent the viruses from entering normal
cells, transductional targeting works by binding "adapter" molecules onto the antibodies, proteins on
the outside of virus, or tweaking the antibodies so that they will not allow viruses to enter normal
cells and rather let them gain up on tumor cells. The second strategy discussed in the article is
named transcriptional targeting. The goal of this form of targeting is to modify the viruses

Proteomics Essay
4.2 Combination of proteomics and interactomics
The study of PPI is fundamental to define the molecular networks that contribute to homeostasis of
living organisms. Disruptions in protein interaction networks have been shown to cause diseases in
both human and animals. For instance, PPI disturbances have been shown to be involved in cancer
caused by p53 mutations [47], neurodegenerative diseases resulting from protein aggregates
accumulation [48], and virus–host interactions [49]. Therefore, the monitoring and study of PPIs can
provide innovative options for identifying diagnostic and therapeutic targets that have potential for
broader clinical applicability. Technological advances in genomics and proteomics have spawned a
large number of ... Show more content on Helpwriting.net ...
Shorter routes to the clinic are also possible because in vitro and in vivo screening, chemical
optimization, toxicology, bulk manufacturing, formulation development and even early clinical
development may have already been completed and can therefore be bypassed. Collectively, these
factors enable several years, and substantial risks and costs, to be removed from the pathway to the
market [51].
By submitting the hub–bottleneck proteins identified through network analysis, we identified
potential drug repositioning candidates. Phosphoaminophosphonic acid–adenylate ester [52], is a
small molecule, classified as a purine ribonucleotide that targets HSP90AA1. It is currently
experimental and has not been evaluated in clinical trials, but is predicted to cross the blood brain
barrier. It may be a promising candidate for treating MPS VII for its anti–oxidant effects. Quercetin,
a small molecule antioxidant targeting ATP5A1, and a naturally occurring molecule, is also a
promising candidate for antioxidant therapy. It is being evaluated in several clinical trials, including
trials evaluating its use for cancer prevention, cancer treatment, and treatment of polyneuropathies.
Rifabutin, which has been shown to also be able to inhibit activation of inflammatory cells and has
demonstrated ability to target HSP90AA1, emerges as another potential drug candidate. Rifabutin is
FDA approved as an

Ortholog Lab Report
Molecular Biology Summative
Question 1:
Orthologs:
These three proteins have been selected as orthologs as they are all proteins from other species
which have very high sequence similarity to the sequence for Homo sapiens dysferlin used. These
sequences have 99%, 99% and 98% identity respectively, all with expectation values of 0.0, so are
very highly conserved between species, with the amino acids that are different generally being very
similar chemically and so will likely carry out the same function as in humans.
Paralogs:
These three proteins have been selected as paralogs for the given protein sequence. There is far
more variation between these compared with the orthologs, as can be seen by the differences in the
alignment scores. The first sequence, myoferlin, has a much lower identity, 56%, but still has a very
high sequence similarity with very few gaps. Myoferlin belongs to the dysferlin sub–family which
the reference sequence is also a part of, with this alignment providing evidence for their
evolutionary closeness.
The second protein, fer–1 like protein 5, has 40% identity and a very high similarity to dysferlin. It
is therefore highly likely to be a ... Show more content on Helpwriting.net ...
There is a high level of sequence homology between the four proteins at this c–terminus end so this
area is highly conserved, however there is quite a bit of variation in regards to which areas are
different. Many of the different amino acids are similar in chemical properties, for example the
variation of P and A where both are neutral and hydrophobic, so it is likely that the protein retained
its function with the different amino acids. However some have more radical differences, for
instance several of the variations are between polar and hydrophobic amino acids. This would have
more of an effect on the proteins as it could cause a change in the structure of the protein and so
change the

Essay On Folding Analysis Of Protein
3.2 Folding analysis of recombinant TGF–β1 protein
To analyze the folding of recombinant TGF–β1, purified protein was electrophoresed on native gel,
under non–reducing condition and stained with the silver stain. Electrophoresis of the purified TGF–
β1, under non–reducing condition, confirmed the presence of dimeric form of the TGF–β1, as a 25
kDa band on the non–denaturing gel (Figure 5 ). Here, there was no trace of a monomeric TGF–β1
band (13.5 kDa). This result indicated that the mature form of TGF–β1 is probably folded properly,
and secreted as a dimer into the culture medium.
3.4 Biological activity of recombinant TGF– β1
The biological activity of purified TGF–β1 was measured in classical growth–inhibition assay on
Mv1–Lu cells. ... Show more content on Helpwriting.net ...
Complicated processes of synthesis, folding and activation cause difficulties to achieve active TGF–
β1 from expression systems. Most previous studies were performed to express the full length of
TGF– β1 in mammalian cells and baculovirus–insect expression systems [11, 21, 22]. However, as
the major secreted proteins by theses cells, consisted of non–covalently associated propeptide with
the mature form of TGF–β1, the yields of biologically active purified proteins were not sufficient,
and significantly decreased after removal of LAP propeptide [11]. Therefore, some attempts were
made to express biologically active TGF– β1, using the DNA sequence, encoding the mature
domain in eukaryotic and prokaryotic systems. However, the expressed proteins were not active,
and in some cases, chemical refolding was required, to maintain the biological activity [9, 10]. Here,
we showed that the expression of biologically active TGF–β1, without the requirement of LAP
coding sequence and chemical refolding, is feasible in yeast expression system. In addition, one
limitation for the accumulation of mature TGF–β1 in the reported expression systems, was the low
stability of the expressed protein, resulted from the activity of endoproteases. To overcome this
problem, we used the engineered PichiaPink, strain 3, knocked out for protease B. Similar to higher
eukaryotes, secreted proteins in yeast can be processed

Nucleic Acid Sequence-Based Amplification ( NASBA )...
Nucleic acid sequence–based amplification (NASBA) assay involves the non–PCR based
amplification of viral RNA of all four dengue virus serotypes using universal primers at 41°C and to
type the amplified products by serotype–specific capture probes by electrochemiluminescence (Wu
et al. 2001). The method is unique in its cost effectiveness, 98.5 % sensitivity, 100 % sensitivity and
short time of one day for investigation (Usawattanakul et al. 2002).
Viral RNA detection using RNA guided clustered regularly interspersed palindromic repeats
(CRISPR) along with Cas13a proteins have been recently developed for the detection of dengue and
zika viruses (Gootenberg et al. 2017). The target viral RNA sequence to be detected is identified
with a ... Show more content on Helpwriting.net ...
2014).The development of a dengue NS1 antigen specific capture enzyme–linked immunosorbent
assay capable of detecting all the four serotypes of dengue virus was tested with patient serum
samples from Tamil Nadu and gave positive results till 7 days of infections (Kathiresan et al. 2017).
Validation of the above results was done with RT–PCR, MAC–ELISA and commercially available
NS1 antigen detection kits. The method was also useful in NS1 antigenic screening in mosquitoes –
the vectors of the disease.
The in silico synthesis and serotyping using multiantigenic peptides of dengue virus proteins as
tools for dengue diagnosis have been recently described (Rai et al. 2017). Epitopes on the envelope
protein and NS1 protein were selected for the synthesis of multiantigenic peptide in the above study.
Results indicated that among 157 patients studied almost 96 % positive reactions against MAP1 for
IgM positive serum and 94 % against MAP 3 for IgM positive serum. However, a reduction in
detection levels were also observed to around 60% IgG positive serums.
Antibody based detection
Antibody based detection of dengue infection can be done using serum samples both for primary
and secondary infections from the 2nd day of infection. The antibody titres

Essay on Biology Case Study
Advanced Cell Biology II Step 1: How will you identify the "vital" cellular protein that the virus
targets for degradation? (Hint: think proteomics). (3 pts.) First, since we know the viral RNA
sequence and its targeting protein, we can investigate it in bioinformatics database, and can acquire
some clues or hints about the target proteins. From the bioinformatics database, we might be able to
find its structure, similarity with other proteins, functions, and binding domains. In other words, we
can get some partial or complete amino sequences of the targeting proteins or information about
likeness. Second, in order to further confirm the information about characteristics and function of
the targeting protein that we have ... Show more content on Helpwriting.net ...
Since we have already known the amino sequence of the protein in previous step, we can narrow
down the targeting ubiquitin ligase based on existing research data such as papers, NCBI data. There
are many types of ubiquitin ligases in cells. However, we can make some candidate groups of
targeting E3 based on the bioinformatics database. We will use antibodies which specifically bind to
each type of ubiquitin ligase and impede its function. For example, the antibodies may covalently
bind to the targeting ubiquitin ligase, and therefore, impede its function. Then, we will measure the
amount of targeting protein. If we find that the amount of targeting protein is not changed in a cell,
we can identify the target ubiquitin ligase. This is because only when the function of the target
ubiquitin ligase is impeded, the degradation of the targeting protein will not be occurred. Step 3:
What protein will be your drug target? What property of that protein will you target? Design an
assay/approach to identify an antidote for "degron". (4 pts.) Since "degron" targets a vital cellular
protein for ubiquitin–dependent degradation, if we block its process of degradation, we can
effectively turn off its effect. As mentioned earlier, ubiquitin ligase brings specificity. Since we have
already known which ubiquitin ligase is

Importance Of Lentiviral Integration Into The Host Genome
Retroviral integration into the host genome is not random and varies dramatically across genera.
Lentiviral HIV–1 has been shown to exhibit strong integration site preferences within active gene
units, whereas gammaretroviral MLV exhibits a strong preference for enhancers and transcription
start sites (Lewinski et al., 2006; Schröder et al., 2002; Wu et al., 2003). These biases have been
attributed to interaction of IN in the context of the pre–integration complex with their cognate host
cell factors (Craigie and Bushman, 2014; Debyser et al., 2015; Kvaratskhelia et al., 2014). For
example, cellular chromatin associated protein lens epithelial–derived growth factor (LEDGF/p75)
interacts with HIV–1 IN and directs lentiviral integration ... Show more content on Helpwriting.net
...
Alpharetroviruses such as ALV exhibit a distinct integration pattern with seemingly random
distribution of integration sites throughout chromatin and with only a slight preference for
integrating into gene regions (Barr et al., 2005; Mitchell et al., 2004; Narezkina et al., 2004;
Withers–Ward et al., 1994). To understand how ALV integration is regulated by host cellular factors,
we have performed affinity capture of the ALV IN protein followed by mass spectrometry (MS)–
based proteomics experiments to identify protein binding partners. Using this approach we
identified structure specific recognition protein 1 (SSRP1) and suppressor of Ty 16 (Spt16), the
components of the heterodimeric FACT (facilitates chromatin transcription) complex (Orphanides et
al., 1999), as the top protein hits that specifically bound to ALV but not HIV–1 IN.
The FACT complex is a highly conserved general histone chaperone protein that is essential for
transcription and DNA replication (Abe et al., 2011; Belotserkovskaya and Reinberg, 2004;
Orphanides et al., 1998). The complex has also been shown to play important roles in DNA damage
responses, centromere deposition, recombination and DNA methylation (Ikeda et al., 2011; Kumari
et al., 2009; Okada et al., 2009; Oliveira et al., 2014). The FACT complex is thought to destabilize

Write An Essay On Rabies Virus
A virus by definition is "any member of a unique class of infectious agents . . . that consists of
genetic material, which may be either DNA or RNA, and is surrounded by a protein coat and, in
some viruses, by a membrane envelope" ("virus"). Such agents are unable to produce the necessary
nutrients to survive because they do not possess the biochemical mechanisms or organelles to
synthesize necessary elements. As a result, they feed off a living host cell by attaching to the cell's
docking proteins and injects its virus through the membrane. However, the cycle of a virus is more
complex than it may seem. There are eight main steps that occur in a basic virus: attachment,
penetration, uncoating, targeting, gene expression, genome replication, virion assembly, and the
release of the new infectious virus. During attachment, the virus's attachment proteins "dock" on
specific receptors found on the outer membrane of certain target cells. In order for this process to
occur, the attachment proteins must bind with a certain receptor for that specific virus. For example,
the rabies virus has glycoprotein attachment molecules that bind with acetylcholine receptors on
neurons. The virus then penetrates the cell and injects its nucleic acid from the capsid, the protein
shell that protects the virus, through either ... Show more content on Helpwriting.net ...
During the sixteenth century, Italian physician Girolamo Fracastoro identified the fatal disease in
humans and discovered that it was caused by a bite from a rabies infested animal and can be
transmitted to other mammals, including humans, through saliva and other bodily fluids. It affects
the mammal's nervous system and causes encephalitis, an inflammation in the brain that can result
in seizures, confusion, unconsciousness, and death. It wasn't until 1885, when a French biologist,
Louis Pasteur, created the first rabies vaccine for

Direct Analytical Sample Quality Assessment (DASQ) for...
There are several important factors to increase the chances that a proteomic study based on CSF
biomarkers becomes successful. For proteomics analysis, the standardisation of CSF collection
protocols is a mandatory step to search and identify new biomarkers indicators of a disease status.
This point represents a milestone especially when samples are collected and stored in different
centres. There is the need to select and include, in biomarker research studies, only well
characterized and high quality samples with a documented sample collection and handling. Our
study undelines the importance of a standardized protocol, based on some specific procedure of
sample collection and storage, in order to have a successful biomarkers analysis. Moreover, it has
been highlighted how MALTI TOF MS analysis of specific proteins could suggest the selection of
the best pool of samples for a powerful biomarker study. We retain that DASQ needs to be used to
screen the samples to be enrolled in order to reduce the number of false positives in biomarker
discovery. Due to the characteristics of MALDI, faster and less expensive than other techniques, this
protocol could represent a first step in the qualitative screening of the CSF before biomarkers
studies. New advanced techniques appear to be particularly promising for biomarker qualification
and verification. Information about these CSF proteins can be used to develop multiple reaction
monitoring (MRM) based analytical assays, which offers

The Effects Of Scorpions
2) Similar to humans, animals have preferences in their diet, whether that be carnivorous,
piscatorial, etc. However, while our food choices, are choices, for most animals, it is innate,
evolutionary or simply has to do with access. For instance, a dessert snake like the inland taipan
wouldn't be piscatorial because there are no fish in the desert. In addition to food choices and
consumption limits, setting is also a hypothesized reason for the uneven distribution of venom
effectiveness in different species. In other words, while venoms and the various types of toxins
within those venoms are going to be harmful to most species, the ones whom are common in their
habitat will be the most effected by their specific toxin. (Casewell, et.al ... Show more content on
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3) Separate file attatched
4) Every species is not venomous or poisonous, instead the inhabitance of toxins belongs to a
special group of individuals who inhabit a toxin or cocktail of toxins they use to protect themselves
and ensure survival. Moreover, as we know, venom is an expensive trait to have and maintain, it is
labor intensive for the species themselves, and they have to have a place to store the venom and an
entirely separate local for administering the venom to their victim. Although venom has proven to
be incredibly advantageous in scribing and eating, it did not just pop up. Instead, the capability
evolved from enzymes and genes carrying out other functions in their bodies (Case well et.al,), these
old functions often directly link to the target of the venom itself where some venoms derive from
mutation with immune system proteins or digestive enzymes. The most common of these mutations
occur in DNA, where DNA is duplicated twice folds as the original, and with that mutation the
original function is unaltered, but the new genes are able to distort the signal and make proteins
elsewhere. Once these mutations take place, natural selection begins to come into play. If these
newly mutated genes begin to create proteins or enzymes populated in the mouth, and these
enzymes get into a prey's womb after it is bit, and the prey is

Advantages And Benefits Of Internet Advertising
There are numerous channels and media that offer you the opportunity to get the best in Internet
advertising strategy.
To understand how it works, how it develops and grows Web advertising is vital.
Then I show my overview of why internet advertising is as effective as a means of promoting your
idea, product, service, project or profession.
My idea is that this will serve as an introduction, you will then interiorizarte and meet more fully the
tools you can implement in your project to achieve the results you want .
Internet advertising – Terminology
To clarify starting from the basics, the Internet advertising also known as Digital Marketing, Internet
Marketing, Web Marketing or Online Marketing.
The term "Internet Advertising or ... Show more content on Helpwriting.net ...
We enumerate the Advantages and Benefits of Internet advertising. effective Internet Free
Newsletters
The costs of Internet advertising are (for now) is generally more economical compared to other
means .
The design and campaign strategy, if well planned, can reach a wide audience (you can advertise to
users anywhere in the world) at a lower price in a very simplified way, accessible to everyone.
The Internet advertising to show off your brand to consumers around the world who buy products
and services physical or virtual at your own pace and convenience.
The Internet Advertising provides detailed statistics that measure more easily and at a much lower
cost all user activity with your brand.
Virtually all elements of an advertising campaign on the Web can be traced and validated in some
way.
L os methods for Internet advertising are revolutionary and offer several alternatives , including pay
per impression (CPM: cost per thousand impressions), pay per click (CPC you pay each time you
click on an advertisement), pay per Views (PPV: pay for video display) or pay per action or
acquisition (PPA: an advertiser pays for a specific action in relation to advertising, such as query
submission form or a

Double-Strand Breaks In Biomedical Research
The past decade has brought rapid and significant innovations in genome–editing techniques. For
the first time researchers have the opportunity to manipulate essentially any gene in a plethora of
cells and organisms, using targeted nucleases that were designed for sequence–specific binding of
the DNA.
One of the first breakthrough methods of gene targeting was the usage of chimeric proteins called
zinc–fingers nucleases (ZFN) to create double–strand breaks. Still, the real revolution was the
introduction of CRISPR and CRISPR associated (Cas) systems into the biomedical research arena.
One specific CRISPR nuclease – Cas9 – paired with short guide RNA has the ability to recognize
the target DNA via Watson–Crick pairing. The guide sequence found ... Show more content on
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One of them is a reported high incidence of nonspecific DNA cleavage; while this has cooled some
of the initial enthusiasm about this method, a potential remedy is the expression of two CRISPR
modules with nickase activity against two genomic sites that are closely adjacent to one another.
Then there is a problem of mosaicism, where mutant allele is produced in only some of the cells, as
nucleases may not inevitably cut the DNA during one stage of embryonic development. The
production of multiple mutations in one organism is also possible, which can create phenotyping
bottlenecks in mouse models.
Regardless of those burning problems, CRISPR/Cas9 genome–editing technique presents staggering
opportunities for addressing a number of illnesses beyond the reach of previous treatment
modalities. Taking into account the accelerating rate of technological progress, as well as a wide
range of research and clinical applications, the road ahead of us will certainly be a thrilling

Electrophoresis: Separation And Analysis Of Macromolecules
Introduction Gel electrophoresis involves separation and analysis of macromolecules like
deoxyribonucleic acid (DNA), ribonucleic acid (RNA), and various proteins. In electrophoresis an
electric current is passed through a solution or gel from one electrode to the other. Ions, molecules,
and molecular fragments in the solution or gel are drawn to one of the electrodes according to their
charge. When charged molecules are placed in a gel, the speed they travel toward the electrode is
influenced by both their charge and size. Molecules can be identified according to which direction
and how fast they travel. (Lab #3 Handout)
Western blot is used to separate and identify proteins by taking mixtures of proteins and separating
them based on molecular ... Show more content on Helpwriting.net ...
The resulting mixtures were set at room temperature for five minutes. These were then poured into
screw cap labeled tubes and heated to 95℃ for another five minutes. While they were being heated,
we created a solution of 1x Tri–Glycine solution equaling 1500 mL out of distilled wter and 10x
Tri–Glycine solution.
Assembly, loading, running, and disassembly of Gel Boxes: Assembly of the Tetra Cell requires
following procedures outlined by the manufacturer's protocol. Once the Tetra cell was assembled,
the solution was added to the chambers of the apparatus. The experiment then requires loading 10
l precision plus protein kaleidoscope prestained into well #2, loading wells #3–7 with the 5 l
prelabeled fish protein solutions into their own wells, and loading well #8 with 10 l of actin &
myosin standard after rising the wells. (See Table 1 in the appendix) The lid was placed on to the
vertical electrophoresis apparatus, set the Bio–Rad power supply to 175V, and then the gel was set
to run. After 45 minutes, all bands were at the bottom of the gel and the Tetra cell was disassembled.
The gel was then placed into a labeled tray with 50 ml of coomassie stain for on hour after being
rinsed with distilled water for five minutes. Once done, the gel was wrapped in plastic

Ribosomes In Eukaryotic Cells
The nucleus houses most of the genes in a eukaryotic cell. The genetic instructions are housed
within a cell, and the instructions are carried out by the ribosomes. The nucleus directs protein
synthesis by synthesizing messenger RNA according to instructions provided by the DNA. The
mRNA is then transported to the cytoplasm via nuclear pores.
The main parts of the nucleus include the nuclear envelope, which encloses the nucleus and
separates it from the cytoplasm, which has an outer and an inner membrane. There is a complex of
pores that helps regulate entry and exit into the nucleus. The nuclear side of the envelope is lined by
the nuclear lamina, which helps maintain the structure of the nucleus. The DNA inside a nucleus is
organized into units called chromosomes, which is made up of chromatin, a complex of proteins and
DNA. The structures in a ... Show more content on Helpwriting.net ...
They carry out protein synthesis. There are two locales– free ribosomes in cytosol and bound ones
to the endoplasmic reticulum or nuclear envelope. Most proteins are made in free ribosomes.
The central dogma of biology is how proteins are made– DNA transcribes RNA, then RNA forms
proteins by translation, or protein synthesis. DNA–>RNA–>Proteins.
Free ribosomes are found in the cytosol, while bound ribosomes are found attached to the side of the
endoplasmic reticulum or nuclear envelope.
The information in a gene,found on a chromosome, in the nucleus is used to synthesize an mRNA
that is transported through a nuclear pore to the cytoplasm. There it is translated into a protein,
which is transported back through a nuclear pore into the nucleus, where it joins other proteins and
DNA, forming chromatin.
The transport vesicles move membranes and substances that they enclose between other components
of the endomembrane system. They are sacs made of membrane that helps transport

Essay On Pyrazole As Therapeutic Agent
In addition to the above uses of pyrazole as therapeutic agent, it has recently been recognized to
have modulatory effect on UPR, especially for the treatment of cancers and other diseases. The
benzyl pyrazole derivative HSF1A, a small molecule activator of HSF–1, was identified in a yeast–
based high–throughput screen (72). Induction of chaperones by HSF1A was shown to reduce protein
misfolding and aggregation–mediated toxicity in cellular and fly models of polyQ–related diseases,
and to activate HSF–1 in Drosophila and mammalian cells without inhibition of Hsp90 activity or
causing proteotoxicity. Rather, HSF1A was suggested to interact with the cytosolic TCP–1 ring
complex (TRiC). This proposed mechanism of action is of interest as TRiC ... Show more content
on Helpwriting.net ...
CNB–001, pyrazole derivative of curcumin for its ability to reduce intracellular Aβ induces eIF2
and PERK (protein kinase R–like extracellular signal–regulated kinase) phosphorylation, and
HSP90 and ATF4 levels. When fed to AD transgenic mice, CNB–001 also increases eIF2
phosphorylation and HSP90 and ATF4 levels, and limits the accumulation of soluble Aβand
ubiquitinated aggregated proteins (80). Pyrazolyl hydroxamic acid derivative, 1–(4–(tert–
Butyl)benzyl)–3–(4–chlorophenyl)–N–hydroxy–1H pyrazole– 5–carboxamide (4f) inhibit Nrf2,
which is persistently activated in many human tumors including AML. s. Treatment with 4f reduced
both B–cell lymphoma–2 (Bcl–2) expression and Bcl–2/Bcl–2–associated X protein (Bax) ratio,
which indicated that 4f induced apoptosis (81).
Ceapins, a class of pyrazole amides have ability to inhibit the processing of ATF6α by S1P and S2P
in response to ER stress. It was demonstrated by probing each step of ATF6α activation that these
ceapins prevent selection of ATF6α into COPII vesicles by retaining it in place in the ER membrane.
In presence of ceapins,it was shown that ATF6α restores the cleavage by removing the requirement
for trafficking that involves bringing together substrate and proteases. Rapid clustering of ATF6α
has been shown to be induced by Ceapins that depicts that the oligomeric state of ATF6α has to play
a key role

Mechanisms Of Ebv Latent Membrane Protein Trafficking Essay
I. TITLE: Mechanisms of EBV Latent Membrane protein trafficking to exosomes
II. BACKGROUND AND SIGNIFICANCE
A. Review of relevant literature
Epstein–Barr–virus (EBV) is a member of the gamma herpesvirus family that establishes a
persistent infection in approximately 90% of the world's population. In immunocompromised
individuals, EBV infection can contribute to cancer development like nasopharyngeal carcinoma,
Burkitt lymphoma and Hodgkin's disease. The latent membrane protein 1 (LMP1) is expressed in
most EBV–associated cancers and it is well established that LMP1 is a major viral oncogene.
Expression of LMP1 alone is sufficient to transform cells and recombinant EBV lacking LMP1 is
incapable of immortalizing B–cells in vitro. Moreover, transgenic mice expressing LMP1 behind a
B–cell specific promoter develop lymphomas.
Exosomes are a population of small (40–150 nm) endocytically–derived extracellular vesicles
produced from inward budding events on the limiting membrane of late endosomal organelles,
forming intraluminal vesicles in multivesicular bodies (MVBs). Epstein Barr virus (EBV) hijacks
the exosomal pathway to modulate cell–to–cell signaling by secretion of viral components such as
LMP1. Trafficking of LMP1 into multivesicular bodies can modify the content and functions of
exosomes. The molecular events orchestrated by LMP1 result in the activation of a plethora of
signaling pathways, including mitogen–activated protein kinase (MAPK/ERK),

Gene And Protein As A Therapeutic Agent For Cancer
In the year 2016, the American Cancer Society projected that 595,690 individuals will pass due to
cancer and 1,685,210 individuals will develop a new cancer case. Many scientists such as Dr. Bert
Vogelstein or Dr. Robert Weinburg have been trying to understand the mechanism behind cancer and
create therapeutic agents that could potentially prevent this disease from occurring. The most novel
and studied gene and protein in regards to cancer is p53. The protein was discovered in the 1970s
when research was focused on cancers that are caused by viruses and was later identified as being a
tumor suppressor (1). One of the most studied areas of cancer is how p53 functions and its role in
the cell cycle which has led to studies that target p53 as a therapeutic agent for cancer (1). In the
year 2001, it was found that the protein product of the gene hSIR2SIRT1, which is a homolog of S.
cerevisiae Sir2 protein, deacetylates the p53 protein and allows for either cell growth arrest or
apoptosis (2). Due to this finding, the Sirtuin 1 protein has been heavily studied and even been
targeted as being a therapeutic target for varying diseases and even involved in the phenomena
known as caloric restriction (3–5). ¬
Current pharmaceutical companies have developed many compounds for cancer treatment that lack
the identification of cellular targets, leading to the absence of cell type specificity in treatment.
However, series of compounds hereafter referred to as de Lijser compounds, have been

Pi3p
Immunoelectron microscopy has shown that PI3P also localizes on the parasite apicoplast and the
food vacuole6. During the asexual blood stage, the Plasmodium endocytoses hemoglobin (the major
cytosolic constituent of red blood cells) and digests it within the food vacuole. This key catabolic
process is largely attenuated in the presence of kinase inhibitors that block PI3P biosynthesis14.
Further investigation into a PI3P–binding protein called FCP in P. falciparum reveals a similar
phenotype (a stunted parasite size and defect in hemoglobin digestion) if its PI3P–binding domain is
deleted15. Again, this finding demonstrates the importance of deciphering PI3P effector proteins in
P. falciparum. These PI3P–regulated functions identified in the ... Show more content on
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An urgent health public concern is raised due to the recent emergence of artemisinin resistance in P.
falciparum17. Artemisinin–based combination therapy is the front–line antimalarial treatment due to
its fast–acting mechanism and high potency against falciparum malaria. Surprisingly, the molecular
target of this potent antimalarial drug has recently been identified to be the PI3–kinase of P.
falciparum18–20. This finding, along with the essentiality and significance of PI3P described above,
accentuates the need to decipher the PI3P–regulated cellular functions in P. falciparum. More
importantly, many Plasmodium proteins are unique to the parasite (e.g., apicoplast enzymes and
FCP), providing excellent targets for drug development. All in all, a more thorough comprehension
of the important PI3P functions on the molecular basis allows for an explicit way to develop
preventive and therapeutic interventions against malaria. Identification of PI3P effector proteins
from a systemic point of view may provide a combinatorial drug therapy and strategy to bypass or
delay the development of drug

Why Chloroplasts Are Important Photosynthetic Organelles?
Chloroplasts are important photosynthetic organelles that present in plant cells. It is believed that
chloroplasts evolve from an endosymbiotic event; engulfment of a photosynthetic cyanobacterium
by a large heterotrophic host cell (1, 2). During this process proteins in the cyanobacterium has been
transferred to the nucleus and also the proteins that are essential for organelle biogenesis has been
transferred to the cyanobacterium making it dependent on the host. Although chloroplast proteins
have estimated to consist of 3500–4000 different types of polypeptides, the protein coding capacity
in chloroplast genes is approximately 200 polypeptides (3, 4). This data further suggest that most of
the proteins found in chloroplast are encode by nuclear genome and transport to the chloroplast. At
least, a few proteins are use secretory pathway in which first targeted to the endoplasmic reticulum
and then transfer to the chloroplast through vesicles (5–7).
Chloroplasts are organized structures for photosynthesis and they consist of three distinct
membranes. They are outer membrane, inner membrane and thylakoid membranes. Photo
complexes, photosynthetic proteins are assemble in the thylakoid membrane network. These three
membranes separate three spaces in the chloroplast, namely, intermembrane space, stroma and
thylakoid lumen. Protein targeting and translocation into chloroplast is therefore of broad interest.
A general outline of chloroplast protein import
According to the

Protein Based Ligands For Tumor Targeting
In addition to peptides, protein–based ligands such as Affibody proteins have been utilized for tumor
targeting. Anti–epidermal growth factor receptor (EGFR) Affibody protein (e.g., Ac–Cys–
ZEGFR:1907, amino acid sequence: Ac–
CVDNKFNKEMWAAWEEIRNLPNLNGWQMTAFIASLVDDPSQSANLLAEAKKLNDAQAPK–
NH2) is used to target EGFR that is overexpressed in a wide variety of human tumors. Cheng and
co–workers used anti–EGFR Affibody protein as tumor–targeting ligand on 64Cu–Au–IO
nanoparticles (PET component: 64Cu, MRI component: iron oxide, specific for EGFR) surface.[63]
In their work, A431 tumor cells were subcutaneously implanted in the right shoulders of nude mice;
and 64Cu–NOTA–Au–IONP–Affibody nanoparticles were administered via tail vain injection.
Rather low resolution PET image showed that these nanoparticles were taken by EGFR positive
A431 (human epithelial carcinoma cell line) cells reaching 4.6% ID/g at 24 h after injection,
significantly higher than that obtained from the blocking experiment (1.9% ID/g) indicating the
specificity of the probe. This difference in % ID/g values corroborated with 44% drop in MRI signal
intensity that was observed for the tumor. In this study, the use of an Affibody protein as ligand for
EGFR–expressing tumor in small animals was demonstrated to be successful; however, the benefit
of multimodality was not demonstrated. The unique chemistry of this dumbbell–shaped Au–IO
nanoprobe could pave the way for targeted drug delivery into EGFR–expressing tumors

Whole Cell Protein Lab Report
Molecules found in nature (termed "natural products") are a rich source of compounds with
biological activities that are useful as drugs. Marine organisms are a particularly good source of
natural products. Imagine that you are a marine biologist searching for new natural products for use
as anticancer drugs. While diving at a coral reef near Tahiti, you notice an unusual sponge that you
have never seen before. You collect a sample, return to your lab and make a whole cell protein
extract.
1. Come up with an experimental design to see if this sponge has anticancer properties.
Making a whole cell protein extract:
Living cells should be cultured on petri dishes and then incubated with a tracer in order for
downstream flux analysis. Once the metabolic processes have been quenched, the next step is to
lyse the cells, separating both the polar and non–polar metabolites from the other cellular substances
at the same time. After these steps, the cells are shaken to completely lyse the membranes allowing
for a more efficient extraction of all possible biomolecules. After shaking, there should be a clear
separation between the polar and non–polar phase for the metabolites, with a well–defined
interphase containing proteins and nucleic acids. After centrifugation, the different phases are now
separated, and each one can be sampled for further analysis ... Show more content on
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During the process of RNA interface long double stranded RNA is cut into small pieces by an
enzyme known as "Dicer". Those small pieces bind with proteins known as the Argonaute proteins.
After this binding takes place one of the double stranded RNA becomes removed which leaves the
remaining strand to bind to messenger RNA target sequences. Once they are bound, the protein can
either recruit certain factors to help regulate it, destroy it, or cleave the messenger

Proper Cell Behavior : The Effects Of Natural Biological...
Proper cell behavior is maintained using a finite pool of processing resources, such as the limited
pool of enzymes required for gene transcription and protein translation1–2. Natural biological
circuits are largely thought to have evolved to buffer against the effects of limited resources, but we
are beginning to understand how processing machinery can form a bottleneck that is in fact
leveraged as a control or signaling mechanism3–4. Proteolytic (protein degrading) pathways, in
particular, have been found to form functional bottlenecks in a native E. coli network regulating the
stationary phase sigma factor S (S). The protein S is degraded by the ClpXP proteolysis system
(ClpXP protease and its chaperones) much faster during ... Show more content on Helpwriting.net ...
1A)8. Queueing theory has since been adopted to describe how competition between substrates for a
particular protease can lead to pronounced coupling and statistical correlation8, 14–16. The impact
of proteolytic queueing competition leads to a rewiring of natural and synthetic circuits to include
mutual modulation of substrate degradation rates17. This effect is due to targeting multiple species
of protein to a common protease ClpXP, and it applies to all but one existing bacterial synthetic
oscillator ClpXP18. The exception is the recently modified repressilator19, where active
degradation by protease was systematically removed to produce a more robust growth–dependent
(dilution–dependent) oscillator, which interestingly was predicted based on a prior analysis of
proteolytic competition17.
The single protease crosstalk picture is too simplistic for native circuits, and the reliance on a single
degradation pathway for bacterial synthetic oscillators presents a scalability problem that limits the
complexity of circuits that can be developed. To address this issue, we investigated the crosstalk
between multiple native degradation pathways in E. coli. This study extends a prior investigation of
computational models that suggested a multi–protease proteolytic bottleneck may still contribute
substantially to crosstalk in simple and complex (oscillatory) networks20. The influence of

Types Of Resistance : Mechanisms Of Antibiotic Resistance
Mechanisms of Resistance
Mechanisms of Antibiotic Resistance
The principle of antibiotic resistance revolve around how antibiotics work. Antibiotics target certain
structures on bacteria such as their cell wall, proteins, and nucleic acids that results in the disruption
and/or inhibition of their growth. These disturbances can sometimes lead to bacterial death. In order
to survive, bacteria have developed countermeasures to fight against the harmful drugs. This was
carried out by targeting the antibiotics themselves. The way antibiotics function is based on their
chemical structure. Because many antibiotics have similar structures, they are also grouped in that
way. Each class (or family) have similarities in their structure and in turn, have similarities in their
target of action. Consequently, these similarities make it easy for the bacteria to construct resistance
to different and multiple classes of antibiotics. Mechanisms that will be discussed all involve
bacteria's ability to prevent antibiotics from reaching its target by means of target alteration, drug
detoxification, impermeability and efflux.
Target Alteration
Target alteration refers to modifications made on the antibiotic molecule. The modification can be
due to point mutations in the genes encoding the target site, enzymes that change binding sites by
acetylation, phosphorylation, adenylation, or replacement/bypass of the original target (1). A good
example of resistance due to point mutation is fluoroquinolone resistance. This drug acts on DNA
gyrase and topoisomerase IV, both of which are vital for bacterial DNA replication. The mutation
lies within the genes that encode for both DNA gyrase and topoisomerase. With its target modified,
fluoroquinolone can no longer inhibit bacterial DNA replication.
Modification can be done chemically by adding certain compounds together or by outright
destroying them. Bacteria employ these actions by producing enzymes that chemically modify
antibiotics by acetylation, phosphorylation, and adenylation (1). This mechanism is seen in both
gram negative and gram positive organisms as with the case of aminoglycoside resistance. This
class of antibiotic is frequently used to treat infections caused by

ERAD In Cystic Fibrosis: A Literature Review
Great Post!
Endoplasmic reticulum–associated degradation (ERAD) seems like a fascinating topic with
catastrophic results that lead to many illnesses. However, what I found interesting is that the role of
ERAD is to serve as the protein quality control system that is generally activated in cases of
neurodegenerative diseases (Elfrink, et al., 2013). In other words, ERAD eliminates misfolded,
damaged, or mutant proteins with abnormal conformation (Vij, et al., 2006). As you mentioned, in
Cystic Fibrosis is the most common disease caused by protein folding mutation known as the cystic
fibrosis transmembrane regulator (CFTR) (Vij, et al., 2006). I had the same thoughts as Dr. Hudry,
with my limit experience in biochemistry and genetics, I assumed if we inhibit ERAD then the
CFTR does not get eliminated, and CF does not exist. However, I know that it is not that simple.
In regards to your questions, some studies touch upon ERAD inhibition as a therapeutic tool that
deals with diseases related to misfolded proteins. In fact, one of the most outstanding studies is titled
"Targeting the ERAD pathway via inhibition of ... Show more content on Helpwriting.net ...
The idea is to use a proteasome inhibitor, in this case, bortezomib, to selectively inhibit Valosin–
Containing Proteins (VCP) that promotes accumulation of immature CFTR in the ER and partial
rescue of functional chloride channels (Vij, Fang, & Zeitlin, 2006). All of this seemed very
complicated, but what they found was that bortezomib could rescue the ΔF508–CFTR from ERAD
and resulted in the appearance of mature CFTR. However, the researchers expressed concern about
using proteasomes as a therapeutic target because proteasomes can be a risk as they are involved in
the generation of various conditions. The biggest challenge is that ERAD has a particular role to
serve as the screener for folded proteins. Therefore, should we inhibit

Leigh's Syndrome
Abstract Leigh's syndrome is caused by a dysfunction in mitochondrial energy generation. It is a
severe disorder that has numerous characteristic features and many symptoms. Mutations in genes
of both the mitochondrial and nuclear DNA can cause Leigh's syndrome; however, a common cause
is as a result of defects in the SURF1 nuclear gene. Functional SURF1 is necessary for the
biogenesis of the cytochrome c oxidase complex, which plays an important role in the electron
transport chain located in the inner mitochondrial membrane. Research is still ongoing to discover
and implement a reliable and effective treatment for Leigh's syndrome.
Introduction
Leigh's syndrome is a rare, progressive mitochondrial and neurodegenerative disorder affecting ...
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Symptomatic treatment aims to improve ATP production and to lower lactate levels. For example,
supplementing the diet with sodium bicarbonate or sodium citrate has proven to control lactic
acidosis. One cause of Leigh's syndrome is pyruvate dehydrogenase deficiency. Thiamine (vitamin
B1), a cofactor of the pyruvate dehydrogenase complex, "has been reported to improve the
neurological status in some patients" (Shrikhande et al. 2010). A promising study demonstrated
"inhibition of the phosphorylation of the pyruvate dehydrogenase complex by thiamine
pyrophosphate, thereby maintaining pyruvate dehydrogenase in the active form" (Hommes et al.
1973). This research showed a therapeutic effect of high doses of thiamine to patients with a
shortage of pyruvate dehydrogenase. A ketogenic (high–fat, adequate–protein, low–carbohydrate)
diet has also been found to improve mitochondrial function and the outcome of individuals with
pyruvate dehydrogenase deficiency. Studies have shown that riboflavin (vitamin B2) may strengthen
mitochondrial function as well (Shrikhande et al.

Db Lab Report
The repair of DSBs
NHEJ initiates when the DSBs is first recognized by the Ku heterodimer, composed of the Ku70 and
Ku80 proteins and binds to the DSB in a sequence–independent manner and in doing so and protects
them from nucleolytic cleavage [33, 34]. The Ku heterodimer then acts as a scaffold to recruits
number NHEJ proteins including DNA–dependent protein kinase (DNA–PKcs) to the DNA ends.
DNA–PKcs is a nuclear protein kinase that phosphorylates a number of protein targets, including
Artemis. Once phosphorylated, Artemis forms an active endonucleolytic complex with DNA–PKcs
that processes the DSB ends to make them compatible for ligation [33, 34]. Pol µ and Pol λ fill in
the DNA gaps and lastly the XRCC4/Ligase IV complex is recruited ... Show more content on
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In the synapsis step, the Rad51–ssDNA filament (presynaptic filament) performs homology search
and DNA strand invasion on a homologous region of another duplex, which results in the formation
of a displacement loop (D–loop) [40]. In the post–synapsis stage of HR the invading 3′ end of the
D–loop primes DNA synthesis by a DNA polymerase, which extends the D–loop enabling the
second 3' single strand terminal end of the DSB to base–pair with it – a process that is called second
end capture. Once captured the second end can itself prime DNA synthesis, and the result of these
"DNA transactions" is the covalent linkage of the recombining DNA molecules via two HJs.
Depending on how the HJs are processed to form mature recombinant products three sub–pathways
of HR are distinguished. In the canonical DSBR model the two HJs (doubleHJ/dHJ) can move along
the DNA by a process called branch migration, which extends or limits the region of DNA
heteroduplex formed by strand exchange catalyzed in the second step. Subsequently, dHJ
intermediate could be resolved by endonucleases such as the Mus81–Eme1/Mms4 complex, Slx1–
Slx4 and the GEN1/Yen1, resulting in the formation of either crossovers (CO) or non–crossover
(NCO) recombinants depending on the orientation in which each HJ is cleaved

The Origin Of The Word Nano
1.1 Introduction
Nanotechnology, the term derived from the Greek word nano, meaning dwarf, applies the principles
of engineering, electronics, physical and material science, and manufacturing at a molecular or
submicron level. The materials at nanoscale could be a device or a system or supramolecular
structures. Earlier Albert Franks defined it as 'that area of science and technology where dimensions
and tolerances are in the nano range. Nanomaterials are the most promising tool in nanotechnology
that posses very unique size dependent properties that makes them superior and indispensable in
many areas of human activity. In recent years, synthesis and characterization of nanoparticles have
received considerable attention because of their distinctive properties and potential uses in various
fields like microelectronics, photocatalysis, magnetic devices, biotechnology and biomedical fields.
Various nanoformulations have already been studied and applied as drug delivery systems with great
success and they still have greater potential for many applications like drugs, antibiotics, protein
delivery, imaging techniques, anti–tumour therapy and as a carrier for Blood Brain Barrier (BBB)
crossing (Yezhelyev, 2006; Chen, 2013; Naahidi, 2013). Nanoparticles provide massive advantages
regarding drug targeting, controlled release and can be combined with diagnosis and other imaging
therapy, hence, emerge as one of the major tools in nanomedicine (Shrivastav, 2013; Jain, 2012).
1.2

PINK1 Genetic Paper
PINK1 (PTEN Induced Putative Kinase 1) is a gene that provides directions for making the protein
PTEN induced putative kinase 1. The greatest amounts of this protein can be found in the muscles,
heart, and testes, but it is also found in cells throughout the body. The protein lies within the
mitochondria of the cell, and, although the function of PTEN induced putative kinase 1 is not yet
fully understood, it seems to assist in protecting the mitochondria from becoming impaired when the
cell is under stress. In order for PTEN induced putative kinase 1 to help the mitochondria, it needs
to first function properly itself. Two specialized regions of this protein ensure that it does this. One
of these regions, the mitochondrial targeting motif, serves as the delivery address. In other words,
once the protein is created, the mitochondrial targeting motif makes sure that it gets to the
mitochondria. The other region, the kinase domain, most likely executes the proteins protective
function. One of the main reasons I chose to research this gene is because of its association with ...
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PINK1, encoded by the PINK1 gene, is a part of "quality control" of the mitochondria It finds
mitochondria that has been damaged and marks them for autophagy; the controlled digestion of an
organelle, in this case, the damaged mitochondria. PINK1 can be taken in and out by healthy
mitochondria because of a membrane potential, but damaged mitochondria lack an adequate
membrane potential to take in PINK1 protein. The protein will then collect on the outer membrane
of the damaged mitochondria, at which point PINK1 will then enlist parkin, another protein
associated with Parkinson's disease, to target the mitochondria for autophagy. The function of
PINK1 is not completely known, but because of it's presence throughout the cytoplasm of cells, a
suggestion of the function of PINK1 is to be a lookout for damaged

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