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By Jamshed Arslan, Pharm. D., PhD.
Protein phosphorylation refers to a reversible post-translational modification in which a protein kinase adds a phosphate group to an amino acid residue of a target protein. Protein phosphorylation, especially tyrosine phosphorylation, is one of the early events in signal transduction in all eukaryotic cells.
Once a cell is lysed, proteases and phosphatases are released that can degrade or...
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By Rachel M.A. Linger, Ph.D.
Survivin is a small (16.5 kDa) protein normally found in human fetal tissue. In contrast, survivin is typically undetectable in most normal adult tissues. Expression of survivin occurs in several subcellular locations including the cytoplasm, nucleus,...
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By Rosa Moreno, PhD.
This new introductory guide to methods and techniques in immunohistochemistry is written by Dr. Alexander E. Kalyuzhny, Bio-Techne, for the Springer series: Techniques in Life Science and Biomedicine for the Non-Expert . This guide provides a comprehensive introduction along with conceptual and methodological basics for new users to ensure a clear understanding upon which researchers may build their expertise in ...
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By Jamshed Arslan, Pharm. D., PhD.
DNA methylation represses transcription of many genes, including tumor suppressor genes. A protein called UHRF1 recruits DNA methyltransferases (DNMTs) to establish and maintain DNA methylation. UHRF1 has several domains, most notably:
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By Michalina Hanzel, PhD
Alexander disease is a progressive and fatal neurological disease with phenotypes ranging from myelination abnormalities, gait ataxia and megalencephaly to predisposition to seizures. It is an autosomal dominant disease with mutations in the glial fibrillary acidic protein (GFAP) gene that result in astrocytic cytoplasmic inclusions, termed Rosenthal fibers, which contribute to the global neurological...
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By Jamshed Arslan, Pharm. D., PhD.
Since its inception in 1979, Western blotting has undergone several developments. The use of radioactive probes was common throughout 1980s, but utilizing secondary antibodies labelled with fluorescent/chemiluminescent probes has virtually removed the need for radioactivity. Western blotting has become a routine quantitative method in biological research and immunodiagnostics. The following primer discusses some considerations when quantitating Western blot data.
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By Jennifer Sokolowski, MD, PhD.
Microglia play important roles in the brain in both homeostatic and pathological conditions, acting to clear debris and dying cells. There is evidence to suggest that microglial dysfunction contributes to neurodegenerative diseases such as Alzheimer's and modulation of microglial activity may be a method to treat such diseases.1 Cell-based therapies represent a novel approach whereby the introduction of microglia that possess the desired phenotype could potentially be...
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By Christina Towers, PhD
Selective autophagic degradation of damaged mitochondria, known as mitophagy, has been described as a cyto-protective process. Accordingly, defects in mitophagy have been associated with a number of diseases including muscle atrophy, cancer, and multiple neurodegenerative diseases. Defective mitophagy has been best described in neurons, where the accumulation of damaged mitochondria and the resulting increase in...
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By Jennifer Sokolowski, MD, PhD.
Human induced pluripotent stem cells (iPSCs) can be used to create models of human organ systems and are useful for a) ascertaining the mechanisms underlying pathological conditions and b) developing and testing therapeutics. For example, studies have used iPSCs from human patients with diseases like Alzheimer's and Parkinson's to test the influence of pathological isoforms of proteins as well as the efficacy of genetic rescue.1,2
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By Jamshed Arslan, Pharm. D., PhD.
Colorectal cancer stem cells are a rare subpopulation of colorectal cancer cells that can self-renew and initiate and sustain tumor growth when transplanted into an animal host.1,2 Colorectal cancer stem cells are identified by CD133, CD166, CD44, nuclear-beta-catenin, ALDH1, EphB2 and...
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By Victoria Osinski
Inflammation results from stimuli signaling damage or infection. The inflammatory response can be beneficial or harmful depending on the type and duration of stimuli. The source, structure, and abundance of these stimuli vary quite a bit. One major category of inflammatory stimulation, or "signal 0s" is the family of pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs).1,2 These patterns are found on bacterial cell walls, DNA, lipoproteins, carbohydrates, or other structures. While many DAMPs and PAMPs have been identified, they stimulate inflammatory...
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By Christina Towers, PhD
Is p62 a good indicator of autophagic flux? The short answer: Yes … but … SQSTM1 encodes the cargo adaptor protein, p62, which interacts with autophagic substrates and delivers them to autophagosomes for degradation. In the process, p62 is itself degraded and when autophagy is induced, a corresponding decrease in p62 levels is observed. Congruently, when autophagy is potently inhibited, i.e. genetic knock out of ATG core autophagy proteins like ATG5 or ATG7, p62 accumulates in the cell. These changes can be observed by immunofluorescent staining, western blotting,...
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By Jamshed Arslan, Pharm. D., PhD.
Cancers in the brain often come from tumors elsewhere in the body. Several adaptive mechanisms influence brain metastasis, such as blood brain barrier leakage that can be induced by stroma or disseminated tumor cells. The brain tumor microenvironment is complex, but the metastasizing cancer cells share common molecular features. To define such molecular characteristics of brain metastasis in vivo, a team led by Dr. Don Nguyen of Yale University School of Medicine developed an advanced RNA sequence-based approach....
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By Michalina Hanzel, PhD
In this final instalment of our three blog-posts series on major cellular mechanisms responsible for neurodegenerative disorders, we will explore the processes of neuroinflammation and microglial activation. Previously, the role of autophagy in the clearance of aggregation-prone proteins in the context of neurodegenerative diseases was discussed in Mechanisms of Neurodegeneration: Protein aggregation and failure of autophagy, and the role of mitochondria and...
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By Christina Towers, PhD
Autophagic flux is defined as the amount of cellular material degraded and recycled through the process of autophagy, whereby cells break down and discard waste. Briefly, the autophagy process involves the formation of an autophagosome, which engulfs cytoplasmic materials. Fusion of the autophagosome with acidic lysosomes mediates the degradation of cellular contents. The process involves over 30 core autophagy proteins (...
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By Beth Melson, MS
Antibiotic resistance is a global threat to public health. Widespread, inappropriate use of antibiotics, such as to treat viral infections or promote growth in livestock, has led to increased incidence of antibiotic resistance1. Resistance is costly to both human health as well as the economy1, and thus the identification of new, effective antibiotics is a public health priority. One solution to this problem is to repurpose commercially available, FDA-approved drugs to target host cell processes that would...
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By Michalina Hanzel, PhD
In this second installment of our three blog-posts series on major cellular mechanisms responsible for neurodegenerative disorders, we will explore the processes of mitochondrial dysfunction and oxidative stress. Previously, the role of autophagy in the clearance of aggregation-prone proteins in the context of neurodegenerative diseases was discussed in Mechanisms of Neurodegeneration: Protein aggregation and failure of autophagy.
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By Jamshed Arslan, Pharm. D., PhD.
Squamous cell carcinoma is the most common cancer in the oral cavity.1 The tumor surface biofilms in oral cancers contain high levels of aerobic and anaerobic microorganisms.1,2 Periodontal pathogens can activate epithelial Toll-like receptors (TLRs) on tumor cells to promote oral squamous cell carcinoma (OSCC) progression. In this regard, TLR2 is known to recognize micro-organisms in the oral cavity by forming a heterodimer with TLR1 or TLR6...
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Kristy R. Howell, PhD
The cellular recycling process known as autophagy may be induced by a variety of conditions including reduced nutrient availability, serum starvation and pharmacological agents (e.g., Rapamycin (mTOR)). Upon autophagy induction, a process of increased sequestration of dysfunctional cellular components into autophagosomes ensues. The...
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By Michalina Hanzel, PhD
In a series of three blog posts I will briefly explore the major cellular mechanisms responsible for many neurodegenerative disorders. The first, and perhaps the most apparent, is the accumulation of misfolded, aggregate-prone proteins, as a result of a failure of the autophagy system, which leads to the loss of neuronal structure and function and eventual neuronal loss.
Autophagy is a conserved cellular process used to eliminate obsolete proteins and cell components by degradation and...
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By Jamshed Arslan, Pharm. D., PhD.
Colon cancer is responsible for over 600,000 deaths per year worldwide. Colon cancer can be classified into two categories: mismatch repair (MMR)-deficient and MMR-proficient cancers.1 MMR-deficient colon cancer shows high CD8+ cytotoxic T-lymphocyte (CTL) and low Th17 cells, while MMR-proficient colon cancer shows an opposite trend. MMR-proficient colon cancer is associated with poor prognosis and is resistant to anti-cancerous immune checkpoint inhibitors. In other words, outcome of immune therapy is...
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By Christina Towers, PhD
Autophagy is a recycling process that relies on the formation of a unique organelle termed an autophagosome. An elegant way to monitor autophagy is through various microscopy techniques to visualize and quantify the formation and turnover of these double membrane structures. Some techniques are more quantitative than others, each accompanied by advantages as well as caveats.
This technique is the only tool that can reveal all of the dynamic stages of a forming...
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By Victoria Osinski
Identifying cellular subsets by flow cytometry requires careful and thorough planning in order to ensure the correct subset of cells are identified and isolated. While generally thought of as antibody-secreting cells, an expanding body of research proves that B cells are a heterogeneous population. Flow cytometry can be used to identify many of these B cell subsets, so long as important nuances and technical approaches are considered.
Before subsetting B cells, steps must be taken to ensure dead cells, cell aggregates, and debris are not...
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By Jamshed Arslan, Pharm. D., PhD.
Atherosclerosis is a chronic inflammatory condition in which plaques of fats and other substances slowly buildup on the inner walls of arteries to restrict blood flow. In atherosclerosis-associated ailments like obesity, hypertension and type II diabetes, intercellular tight junctions are impaired, leading to increased gut permeability. This means that products of intestinal microbiota like lipopolysaccharide from Gram-negative bacteria can seep into the blood stream.
A pro-atherogenic role of lipopolysaccharide has been suggested. For example,...
![Expression of RhoA, B and C detected in the cytoplasm of A431 cells, a model of human epidermoid carcinoma, compared to negative control analyzed in the absence of RhoA Antibody (1A11-4G10) [NBP2-22528]](https://images.novusbio.com/design/RhoA-B-C-expression-A431-cells-header.jpg "RhoA, B and C expression in A431 cells, ICC")