The document summarizes a study that investigated whether the HIV-1 Nef gene enhances the production of beta amyloid, a protein involved in Alzheimer's disease. Mouse brain cells were transfected with the Nef gene, and exosomes produced were collected and applied to untreated brain cells. ELISA and Western blot tests found that Nef exosomes increased the secretion of beta amyloid peptides from treated cells, supporting the hypothesis. Further research on human cells was proposed to explore implications for HIV-associated dementia.
Prevalence of Staphylococcal enterotoxin genes in Italy - Guerrino Macori inv...Guerrino Macori
This document summarizes the results of a study on the prevalence of staphylococcal enterotoxin genes in Italy. 1245 dairy samples were collected and tested for 11 staphylococcal enterotoxin genes. 255 samples (20%) were positive for at least one gene. The most common genes were ser, sed, and selj. Further analysis found that sed-ser-selj was the most common genetic profile, found in 15% of samples, followed by seg-sei in 12.3% of samples. Many of these genes are often found together because they are located within the same genetic clusters or on the same plasmids. The results were consistent with previous studies and data from other European countries. Further
This study examined apoptosis of lymphocytes in patients with systemic lupus erythematosus (SLE). The percentage of apoptotic lymphocytes from SLE patients was significantly higher than healthy donors after 36 hours of incubation, indicating SLE patients have more lymphocytes undergoing apoptosis. A positive correlation was also found between lymphocyte apoptosis in SLE and disease activity markers like anti-dsDNA antibodies and prednisolone dosage. The percentage of CD4+ T cells was significantly lower in SLE patients compared to healthy donors.
Enhancing Solubilization of Hydrophobic ORF3 Product of HEV in Bacterial Expr...Allyson Luo
This study aimed to enhance the expression and solubility of the vp13 protein from hepatitis E virus (HEV) by expressing it as a fusion protein with maltose binding protein (MBP) in E. coli. The researchers cloned the ORF3 gene encoding vp13 into a bacterial expression vector with an MBP tag. Initial expression trials found most of the MBP-vp13 fusion protein was insoluble. The researchers then optimized induction conditions, finding induction at 18°C and addition of 1% glucose led to efficient expression of soluble MBP-vp13 fusion protein. This enhanced solubilization and purification of vp13, which will facilitate further study of its structure and function.
The researchers developed a novel non-viral single doxycycline inducible system that can efficiently regulate gene expression in stem cells both in vitro and in vivo. They generated stable clonal cell lines from mouse neuroblastoma cells (N2a) and human embryonic stem cells (hNSC-ESC) that responded to doxycycline by eliminating GFP expression within 3-8 days. GFP expression was restored 9-10 days after doxycycline removal. When transplanted hNSC-ESCs labeled with quantum dots were subjected to doxycycline in mice, GFP expression was reduced by 61-56% compared to controls. This suggests the system can control gene expression in stem cells without using viral vectors
- The epithelium in the cauda epididymis of male mice lacking the GPR56 gene is severely thinner than in wildtype mice and shows lesions in the basement membrane.
- The lumen of the cauda epididymis in GPR56 knockout mice contains few sperm and many residual bodies, while the wildtype lumen is filled with sperm.
- Abnormalities in the epithelium and lumen of the cauda epididymis in GPR56 knockout mice may affect their fertility, as the cauda is where sperm are stored.
Molecular Identification of Specific Virulence Genes in EnteropathogenicEsche...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
This document discusses various applications of stem cell research and therapy. It summarizes that stem cells can differentiate into specialized cell types, cell therapy uses healthy live or freeze-dried cells to treat diseases, and stem cells have been used in animal models to treat conditions like arthritis and Parkinson's disease. Stem cell research also aims to produce tissues for transplantation through cloning cells and growing them on scaffolds.
The prevalence of Copy Number Variation (CNV) on human chromosome 16p11.2, identified in approximately 1% of autism spectrum disorder (ASD) cases globally, was found to be 1.2% in an Australian ASD cohort. Bioinformatic analysis identified 13 of the 25 protein coding genes within the 16p11.2 region, including KCTD13, as significantly enriched in the nervous system. Experiments in mice found that suppression of Kctd13 led to impaired radial migration of cortical neurons and altered neuronal morphology, providing insight into how perturbations to KCTD13 expression via 16p11.2 microdeletion could influence brain development. Further experiments showed decreased cell proliferation and mitosis levels
Prevalence of Staphylococcal enterotoxin genes in Italy - Guerrino Macori inv...Guerrino Macori
This document summarizes the results of a study on the prevalence of staphylococcal enterotoxin genes in Italy. 1245 dairy samples were collected and tested for 11 staphylococcal enterotoxin genes. 255 samples (20%) were positive for at least one gene. The most common genes were ser, sed, and selj. Further analysis found that sed-ser-selj was the most common genetic profile, found in 15% of samples, followed by seg-sei in 12.3% of samples. Many of these genes are often found together because they are located within the same genetic clusters or on the same plasmids. The results were consistent with previous studies and data from other European countries. Further
This study examined apoptosis of lymphocytes in patients with systemic lupus erythematosus (SLE). The percentage of apoptotic lymphocytes from SLE patients was significantly higher than healthy donors after 36 hours of incubation, indicating SLE patients have more lymphocytes undergoing apoptosis. A positive correlation was also found between lymphocyte apoptosis in SLE and disease activity markers like anti-dsDNA antibodies and prednisolone dosage. The percentage of CD4+ T cells was significantly lower in SLE patients compared to healthy donors.
Enhancing Solubilization of Hydrophobic ORF3 Product of HEV in Bacterial Expr...Allyson Luo
This study aimed to enhance the expression and solubility of the vp13 protein from hepatitis E virus (HEV) by expressing it as a fusion protein with maltose binding protein (MBP) in E. coli. The researchers cloned the ORF3 gene encoding vp13 into a bacterial expression vector with an MBP tag. Initial expression trials found most of the MBP-vp13 fusion protein was insoluble. The researchers then optimized induction conditions, finding induction at 18°C and addition of 1% glucose led to efficient expression of soluble MBP-vp13 fusion protein. This enhanced solubilization and purification of vp13, which will facilitate further study of its structure and function.
The researchers developed a novel non-viral single doxycycline inducible system that can efficiently regulate gene expression in stem cells both in vitro and in vivo. They generated stable clonal cell lines from mouse neuroblastoma cells (N2a) and human embryonic stem cells (hNSC-ESC) that responded to doxycycline by eliminating GFP expression within 3-8 days. GFP expression was restored 9-10 days after doxycycline removal. When transplanted hNSC-ESCs labeled with quantum dots were subjected to doxycycline in mice, GFP expression was reduced by 61-56% compared to controls. This suggests the system can control gene expression in stem cells without using viral vectors
- The epithelium in the cauda epididymis of male mice lacking the GPR56 gene is severely thinner than in wildtype mice and shows lesions in the basement membrane.
- The lumen of the cauda epididymis in GPR56 knockout mice contains few sperm and many residual bodies, while the wildtype lumen is filled with sperm.
- Abnormalities in the epithelium and lumen of the cauda epididymis in GPR56 knockout mice may affect their fertility, as the cauda is where sperm are stored.
Molecular Identification of Specific Virulence Genes in EnteropathogenicEsche...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
This document discusses various applications of stem cell research and therapy. It summarizes that stem cells can differentiate into specialized cell types, cell therapy uses healthy live or freeze-dried cells to treat diseases, and stem cells have been used in animal models to treat conditions like arthritis and Parkinson's disease. Stem cell research also aims to produce tissues for transplantation through cloning cells and growing them on scaffolds.
The prevalence of Copy Number Variation (CNV) on human chromosome 16p11.2, identified in approximately 1% of autism spectrum disorder (ASD) cases globally, was found to be 1.2% in an Australian ASD cohort. Bioinformatic analysis identified 13 of the 25 protein coding genes within the 16p11.2 region, including KCTD13, as significantly enriched in the nervous system. Experiments in mice found that suppression of Kctd13 led to impaired radial migration of cortical neurons and altered neuronal morphology, providing insight into how perturbations to KCTD13 expression via 16p11.2 microdeletion could influence brain development. Further experiments showed decreased cell proliferation and mitosis levels
Click Colorimetric EdU Proliferation and TUNEL: Click Chemistry for Bright Fi...Thermo Fisher Scientific
We present the conversion of click chemistry EdU and TUNEL fluorescence assay to colorimetric detection of proliferation and apoptosis for use with bright field microscopy. Advantages of having a click colorimetric signal are retaining the work flow of click chemistry labeling while allowing for compatibility with standard H&E staining and protocols.
This document describes a study evaluating the use of primary neuronal cultures to test the efficacy of antiprion drugs. The researchers were able to propagate different prion strains from various species, including sheep, mouse, hamster and human, in primary neuronal cultures derived from transgenic mouse lines overexpressing the corresponding prion protein. Using this system, they tested three generic antiprion compounds and found their activity was dependent on the prion strain and species, recapitulating some in vivo results. The study suggests infected primary neuronal cultures may be a relevant model for investigating antiprion drug efficacy and mechanisms of action.
Regulation of embryonic stem cell pluripotency by heatMahmoud El Beblawy
Hsp90 maintains embryonic stem cell pluripotency by regulating key pluripotency factors like Oct4 and Nanog. Inhibition of Hsp90 using drugs like 17-AAG in mouse embryonic stem cells leads to:
1) Changes in cell morphology from dome shaped to flattened.
2) Loss of alkaline phosphatase staining and pluripotent colony formation.
3) Decreased levels of Oct4, Nanog and phosphorylated Stat3.
4) Reduced teratoma formation in mice.
ELISA Vs ELISPOT - Principle, Procedure, Advantagesajithnandanam
The Enzyme Linked Immunospot (ELISPOT) technique was developed by Cecil Czerkinskdy in 1983. ELISPOT is used for the detection of secreted proteins, such as cytokines and growth factors. ELISPOT is primarily used in immunology research in the following areas:
1) The study analyzed the effects of Csk knockouts on development of the initial segment of the mouse epididymis. Csk knockout was expected to promote cell proliferation and differentiation through increased ERK pathway activity due to lack of inhibition of SRC kinases.
2) A tissue-specific Csk conditional knockout mouse model was generated using Cre/lox recombination. Genotyping identified one mouse with the desired genotype.
3) Preliminary results found increased vasculogenesis in the initial segment of Csk knockout mice, suggesting effects on differentiation through the ERK pathway. Immunofluorescence found decreased activity of phospho-SRC in knockouts while other markers were similar to controls.
1) A study observed increased OPG levels in the feces and biopsy cultures of children with inflammatory bowel disease compared to non-IBD controls. Immunostaining of biopsies showed OPG signal in goblet cells of IBD patients.
2) Experiments with intestinal epithelial cell lines found nuclear OPG expression and secretion increased with TNF-α and IL-1β treatment. Knockdown of OPG via siRNA decreased OPG secretion but did not affect IL-8 levels.
3) Future studies aim to investigate the role of OPG in intestinal epithelial cell-bacteria interactions and overexpression of OPG-GFP in intestinal epithelial cells.
1) Cultured human limbal epithelial cells were stored for one week at 31°C in organ culture medium, 23°C in organ culture medium, or 5°C in Optisol-GS. Cell death due to apoptosis was assessed using caspase-3 staining, TUNEL staining, and gene expression profiling of 84 apoptosis genes.
2) Minimal cell death was observed under all storage conditions, as indicated by low caspase-3 and TUNEL labeling. Gene expression analysis found pro-apoptotic genes were downregulated and anti-apoptotic genes were upregulated.
3) Organ culture storage at 23°C best preserved cell structure, while 31°C and 5°C storage were associated
The document provides information about midterm exams and extra office hours for a class. It announces that the first midterm exam will be on Thursday, February 15 at 6pm in room 155 Dwinelle, and that review sessions will be held during the regular class time that day. It also notes that the professor will not have office hours on February 13.
Quorum sensing allows bacteria to communicate and coordinate behaviors based on cell population density. Gram negative bacteria use acyl-homoserine lactones and Gram positive bacteria use oligopeptides to signal each other. Quorum sensing regulates biofilm formation in many pathogenic bacteria, including Pseudomonas aeruginosa, Staphylococcus aureus, Vibrio cholerae, and Listeria monocytogenes. It controls production of virulence factors and formation of mature biofilms at high cell densities. Mutations disrupting quorum sensing decrease biofilm formation in several bacteria.
1. Dr. Madhuri Hegde presented on detecting small intragenic deletions using targeted comparative genomic hybridization (aCGH).
2. She discussed several examples of deletions less than 2.5 kb detected by aCGH in disease genes including PAH, STK11, HPRT1, and EMD.
3. She concluded that aCGH is a valuable tool for detecting small intragenic deletions and providing insights into deletion mechanisms.
Targeted and Unbiased Screen for Genetic Suppressors of the Legionella pneumo...Chetan Hebbale
This document describes research into identifying genetic suppressors of toxicity caused by the Legionella pneumophila effector protein LegC7 in Saccharomyces cerevisiae. A targeted screen of Class E vacuolar protein sorting gene deletions did not suppress LegC7 toxicity except for vps27Δ. An unbiased ethyl methanesulfonate mutagenesis screen identified yeast strains where LegC7 toxicity was reversed, likely due to genomic or plasmid mutations. The genomes of two strains will be sequenced to find specific mutations, and an overexpression screen will search for genes that reduce LegC7 toxicity when overexpressed.
This study developed a sensitive sandwich enzyme-linked immunosorbent assay (ELISA) to detect tau proteins in cerebrospinal fluid (CSF) using the monoclonal antibody AT120. The assay had a detection limit of less than 5 pg/ml of CSF for tau. Using this assay, CSF tau levels were found to be significantly higher in Alzheimer's disease patients compared to controls. However, CSF tau levels overlapped between Alzheimer's patients and those with other neurological diseases. The study suggests CSF tau measurement may help diagnose Alzheimer's but cannot on its own distinguish it from other conditions.
The Influence of Various Drags on Mortality of Mice and the Concentration of ...CrimsonPublishersAICS
The Influence of Various Drags on Mortality of Mice and the Concentration of Proinflammatory Cytokines in Blood at Sepsis Caused by E. Coli by Zabrodskii PF* in Advancements in Case Studies
PREVALENCE AND ANTIMICROBIAL SUSCEPTIBILITY OF ESBL IN SOKOTO PDFNuhu Tanko
This study examined the prevalence and antimicrobial susceptibility of extended-spectrum beta-lactamase (ESBL)-producing gram-negative uropathogens in Sokoto, Nigeria. 365 urine samples were collected and analyzed between November 2014 and February 2015. Gram-negative uropathogens made up 60.9% of positive cultures. The most common isolates were E. coli (29.7%) and Salmonella arizonae (23.4%). 15 isolates (23.3%) were confirmed as ESBL producers, with E. coli and Enterobacter gergoviae being the most common. The isolates showed high resistance to cotrimoxazole but high susceptibility to nitrofurantoin. This study demonstrates
Apoptosis is a programmed cell death process that occurs in multicellular organisms. It is characterized by cell shrinkage, nuclear fragmentation, and global mRNA decay. Apoptosis ensures the orderly disposal of cells and occurs during normal cell turnover, development, and aging. It also helps remove infected, damaged or unnecessary cells. There are two main pathways of apoptosis - the intrinsic pathway which is triggered by intracellular signals within the cell, and the extrinsic pathway which is initiated by activation of cell surface death receptors by extracellular ligands. Both pathways activate caspases, a family of cysteine proteases, that trigger the degradation of proteins and DNA fragmentation that leads to cell death. Deregulation of apoptosis can contribute to diseases like cancer
This document describes a protocol for differentiating mouse induced pluripotent stem cells (miPSCs) into oligodendrocyte progenitor cells (OPCs). The protocol utilizes a defined, stepwise differentiation process over 8 days to generate high yields of OPCs from three different miPSC lines. The OPCs produced from this protocol are capable of further differentiation into mature, myelinating oligodendrocytes both in vitro and in vivo, making them a potentially useful cell source for modeling demyelinating disorders and developing cell-based therapies.
- The document discusses neuronal cell death in Alzheimer's disease, which is linked to two protein aggregates - amyloid beta plaques and tau tangles. When amyloid beta is introduced to healthy mouse neurons in culture, it leads to rapid cell death.
- Amyloid beta treatment of the neurons increases the activity of calcium-dependent proteases (calpains) and cysteine proteases (caspases), which quickly degrade tau through cleavage into lower molecular weight fragments. This degradation of tau by proteases appears to contribute to neuronal cell death.
- The phosphorylation of tau by kinases such as GSK3 beta and CDK5 also increases after amyloid beta treatment, further implicating tau changes in the neuronal death caused by amyloid
This study examined the expression of endothelial nitric oxide synthase (eNOS) in testicular tissue from men with normal spermatogenesis (control group) and men with non-obstructive azoospermia (NOA group). Immunohistochemistry revealed that eNOS was present in Leydig cells, Sertoli cells, immature spermatids and abnormal germ cells in both groups. However, eNOS expression was significantly higher in Leydig cells of the control group and higher in Sertoli cells and abnormal germ cells of the NOA group. The higher eNOS expression in the NOA group suggests it may play a role in the apoptosis of abnormal germ cells and the disruption of sper
UTF-8''Final Assessing post-synaptic partners of Dentate Granule Cells in a M...Grant Pizzo
1) The document describes an experiment assessing how dentate granule cell (DGC) axon terminals and their post-synaptic partners change in a rat model of temporal lobe epilepsy (TLE).
2) The researcher induced seizures in rats using pilocarpine and later injected retroviruses to label dividing DGCs. Tissue was then analyzed to identify post-synaptic partners of DGCs using antibodies targeting interneurons and mossy cells.
3) Preliminary results showed that antibodies for GAD67 and GluR2 successfully labeled the correct cell types, but need further refinement, while an mGluR1/5 antibody did not label interneurons in the dentate gyrus
Detailed steps on how to achieve beautiful skin whether you're concerned with acne, anti-aging, fine lines, wrinkles, dark spots, hyper pigmentation, age spots, dark circles and the best skin care products for beautiful skin. Our clinic is in Mesa, Arizona in the Phoenix Area, but we offer a long-distance program anywhere in the United States.
Electus Recruitment Solutions is a recruitment firm specializing in cyber security and information assurance talent. They work with clients in aerospace, defense, energy, and infrastructure. Their goal is to quickly find and secure highly skilled talent through sophisticated search methods and dedicated consultant teams with expertise in specific sectors. Electus offers various talent solutions tailored to clients' needs, from contingency to retained recruitment services. They take pride in their culture of developing expert consultants to deliver top quality service and the best talent.
Click Colorimetric EdU Proliferation and TUNEL: Click Chemistry for Bright Fi...Thermo Fisher Scientific
We present the conversion of click chemistry EdU and TUNEL fluorescence assay to colorimetric detection of proliferation and apoptosis for use with bright field microscopy. Advantages of having a click colorimetric signal are retaining the work flow of click chemistry labeling while allowing for compatibility with standard H&E staining and protocols.
This document describes a study evaluating the use of primary neuronal cultures to test the efficacy of antiprion drugs. The researchers were able to propagate different prion strains from various species, including sheep, mouse, hamster and human, in primary neuronal cultures derived from transgenic mouse lines overexpressing the corresponding prion protein. Using this system, they tested three generic antiprion compounds and found their activity was dependent on the prion strain and species, recapitulating some in vivo results. The study suggests infected primary neuronal cultures may be a relevant model for investigating antiprion drug efficacy and mechanisms of action.
Regulation of embryonic stem cell pluripotency by heatMahmoud El Beblawy
Hsp90 maintains embryonic stem cell pluripotency by regulating key pluripotency factors like Oct4 and Nanog. Inhibition of Hsp90 using drugs like 17-AAG in mouse embryonic stem cells leads to:
1) Changes in cell morphology from dome shaped to flattened.
2) Loss of alkaline phosphatase staining and pluripotent colony formation.
3) Decreased levels of Oct4, Nanog and phosphorylated Stat3.
4) Reduced teratoma formation in mice.
ELISA Vs ELISPOT - Principle, Procedure, Advantagesajithnandanam
The Enzyme Linked Immunospot (ELISPOT) technique was developed by Cecil Czerkinskdy in 1983. ELISPOT is used for the detection of secreted proteins, such as cytokines and growth factors. ELISPOT is primarily used in immunology research in the following areas:
1) The study analyzed the effects of Csk knockouts on development of the initial segment of the mouse epididymis. Csk knockout was expected to promote cell proliferation and differentiation through increased ERK pathway activity due to lack of inhibition of SRC kinases.
2) A tissue-specific Csk conditional knockout mouse model was generated using Cre/lox recombination. Genotyping identified one mouse with the desired genotype.
3) Preliminary results found increased vasculogenesis in the initial segment of Csk knockout mice, suggesting effects on differentiation through the ERK pathway. Immunofluorescence found decreased activity of phospho-SRC in knockouts while other markers were similar to controls.
1) A study observed increased OPG levels in the feces and biopsy cultures of children with inflammatory bowel disease compared to non-IBD controls. Immunostaining of biopsies showed OPG signal in goblet cells of IBD patients.
2) Experiments with intestinal epithelial cell lines found nuclear OPG expression and secretion increased with TNF-α and IL-1β treatment. Knockdown of OPG via siRNA decreased OPG secretion but did not affect IL-8 levels.
3) Future studies aim to investigate the role of OPG in intestinal epithelial cell-bacteria interactions and overexpression of OPG-GFP in intestinal epithelial cells.
1) Cultured human limbal epithelial cells were stored for one week at 31°C in organ culture medium, 23°C in organ culture medium, or 5°C in Optisol-GS. Cell death due to apoptosis was assessed using caspase-3 staining, TUNEL staining, and gene expression profiling of 84 apoptosis genes.
2) Minimal cell death was observed under all storage conditions, as indicated by low caspase-3 and TUNEL labeling. Gene expression analysis found pro-apoptotic genes were downregulated and anti-apoptotic genes were upregulated.
3) Organ culture storage at 23°C best preserved cell structure, while 31°C and 5°C storage were associated
The document provides information about midterm exams and extra office hours for a class. It announces that the first midterm exam will be on Thursday, February 15 at 6pm in room 155 Dwinelle, and that review sessions will be held during the regular class time that day. It also notes that the professor will not have office hours on February 13.
Quorum sensing allows bacteria to communicate and coordinate behaviors based on cell population density. Gram negative bacteria use acyl-homoserine lactones and Gram positive bacteria use oligopeptides to signal each other. Quorum sensing regulates biofilm formation in many pathogenic bacteria, including Pseudomonas aeruginosa, Staphylococcus aureus, Vibrio cholerae, and Listeria monocytogenes. It controls production of virulence factors and formation of mature biofilms at high cell densities. Mutations disrupting quorum sensing decrease biofilm formation in several bacteria.
1. Dr. Madhuri Hegde presented on detecting small intragenic deletions using targeted comparative genomic hybridization (aCGH).
2. She discussed several examples of deletions less than 2.5 kb detected by aCGH in disease genes including PAH, STK11, HPRT1, and EMD.
3. She concluded that aCGH is a valuable tool for detecting small intragenic deletions and providing insights into deletion mechanisms.
Targeted and Unbiased Screen for Genetic Suppressors of the Legionella pneumo...Chetan Hebbale
This document describes research into identifying genetic suppressors of toxicity caused by the Legionella pneumophila effector protein LegC7 in Saccharomyces cerevisiae. A targeted screen of Class E vacuolar protein sorting gene deletions did not suppress LegC7 toxicity except for vps27Δ. An unbiased ethyl methanesulfonate mutagenesis screen identified yeast strains where LegC7 toxicity was reversed, likely due to genomic or plasmid mutations. The genomes of two strains will be sequenced to find specific mutations, and an overexpression screen will search for genes that reduce LegC7 toxicity when overexpressed.
This study developed a sensitive sandwich enzyme-linked immunosorbent assay (ELISA) to detect tau proteins in cerebrospinal fluid (CSF) using the monoclonal antibody AT120. The assay had a detection limit of less than 5 pg/ml of CSF for tau. Using this assay, CSF tau levels were found to be significantly higher in Alzheimer's disease patients compared to controls. However, CSF tau levels overlapped between Alzheimer's patients and those with other neurological diseases. The study suggests CSF tau measurement may help diagnose Alzheimer's but cannot on its own distinguish it from other conditions.
The Influence of Various Drags on Mortality of Mice and the Concentration of ...CrimsonPublishersAICS
The Influence of Various Drags on Mortality of Mice and the Concentration of Proinflammatory Cytokines in Blood at Sepsis Caused by E. Coli by Zabrodskii PF* in Advancements in Case Studies
PREVALENCE AND ANTIMICROBIAL SUSCEPTIBILITY OF ESBL IN SOKOTO PDFNuhu Tanko
This study examined the prevalence and antimicrobial susceptibility of extended-spectrum beta-lactamase (ESBL)-producing gram-negative uropathogens in Sokoto, Nigeria. 365 urine samples were collected and analyzed between November 2014 and February 2015. Gram-negative uropathogens made up 60.9% of positive cultures. The most common isolates were E. coli (29.7%) and Salmonella arizonae (23.4%). 15 isolates (23.3%) were confirmed as ESBL producers, with E. coli and Enterobacter gergoviae being the most common. The isolates showed high resistance to cotrimoxazole but high susceptibility to nitrofurantoin. This study demonstrates
Apoptosis is a programmed cell death process that occurs in multicellular organisms. It is characterized by cell shrinkage, nuclear fragmentation, and global mRNA decay. Apoptosis ensures the orderly disposal of cells and occurs during normal cell turnover, development, and aging. It also helps remove infected, damaged or unnecessary cells. There are two main pathways of apoptosis - the intrinsic pathway which is triggered by intracellular signals within the cell, and the extrinsic pathway which is initiated by activation of cell surface death receptors by extracellular ligands. Both pathways activate caspases, a family of cysteine proteases, that trigger the degradation of proteins and DNA fragmentation that leads to cell death. Deregulation of apoptosis can contribute to diseases like cancer
This document describes a protocol for differentiating mouse induced pluripotent stem cells (miPSCs) into oligodendrocyte progenitor cells (OPCs). The protocol utilizes a defined, stepwise differentiation process over 8 days to generate high yields of OPCs from three different miPSC lines. The OPCs produced from this protocol are capable of further differentiation into mature, myelinating oligodendrocytes both in vitro and in vivo, making them a potentially useful cell source for modeling demyelinating disorders and developing cell-based therapies.
- The document discusses neuronal cell death in Alzheimer's disease, which is linked to two protein aggregates - amyloid beta plaques and tau tangles. When amyloid beta is introduced to healthy mouse neurons in culture, it leads to rapid cell death.
- Amyloid beta treatment of the neurons increases the activity of calcium-dependent proteases (calpains) and cysteine proteases (caspases), which quickly degrade tau through cleavage into lower molecular weight fragments. This degradation of tau by proteases appears to contribute to neuronal cell death.
- The phosphorylation of tau by kinases such as GSK3 beta and CDK5 also increases after amyloid beta treatment, further implicating tau changes in the neuronal death caused by amyloid
This study examined the expression of endothelial nitric oxide synthase (eNOS) in testicular tissue from men with normal spermatogenesis (control group) and men with non-obstructive azoospermia (NOA group). Immunohistochemistry revealed that eNOS was present in Leydig cells, Sertoli cells, immature spermatids and abnormal germ cells in both groups. However, eNOS expression was significantly higher in Leydig cells of the control group and higher in Sertoli cells and abnormal germ cells of the NOA group. The higher eNOS expression in the NOA group suggests it may play a role in the apoptosis of abnormal germ cells and the disruption of sper
UTF-8''Final Assessing post-synaptic partners of Dentate Granule Cells in a M...Grant Pizzo
1) The document describes an experiment assessing how dentate granule cell (DGC) axon terminals and their post-synaptic partners change in a rat model of temporal lobe epilepsy (TLE).
2) The researcher induced seizures in rats using pilocarpine and later injected retroviruses to label dividing DGCs. Tissue was then analyzed to identify post-synaptic partners of DGCs using antibodies targeting interneurons and mossy cells.
3) Preliminary results showed that antibodies for GAD67 and GluR2 successfully labeled the correct cell types, but need further refinement, while an mGluR1/5 antibody did not label interneurons in the dentate gyrus
Detailed steps on how to achieve beautiful skin whether you're concerned with acne, anti-aging, fine lines, wrinkles, dark spots, hyper pigmentation, age spots, dark circles and the best skin care products for beautiful skin. Our clinic is in Mesa, Arizona in the Phoenix Area, but we offer a long-distance program anywhere in the United States.
Electus Recruitment Solutions is a recruitment firm specializing in cyber security and information assurance talent. They work with clients in aerospace, defense, energy, and infrastructure. Their goal is to quickly find and secure highly skilled talent through sophisticated search methods and dedicated consultant teams with expertise in specific sectors. Electus offers various talent solutions tailored to clients' needs, from contingency to retained recruitment services. They take pride in their culture of developing expert consultants to deliver top quality service and the best talent.
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive functioning. Exercise causes chemical changes in the brain that may help protect against mental illness and improve symptoms.
Michelle Ulman has over 20 years of experience in business development, customer relations, and nonprofit leadership. She has a track record of growing organizations through networking, marketing, and partnership building. Ulman holds a degree in business administration and has received several honors for her work developing local economies and communities.
Sajja Girikumar has over 7 years of experience in software development with a focus on ETL using Ab Initio. He has extensive experience developing ETL processes and data integration solutions for clients in the healthcare and banking industries. Currently he works as an onsite team lead for Tetrasoft in Little Rock, Arkansas, managing teams of up to 8 members.
We presented our film opening to classmates for feedback. They suggested adding ambient sounds at the beginning to set the atmosphere. Some drone shots came and went too quickly, making parts feel jumpy and out of sync like when a character returned to a car. However, crosscutting between shots and car scenes worked well and were quick. For the ending phone call scene, the voice was too loud to realistically come from a phone, so it needs to be quieted and modified to sound like an actual phone call. Overall the opening captured the quick pace of a kidnapping situation realistically.
This document summarizes a collaborative meeting for teacher assistants and aides led by Derek Warren on February 4th, 2016. The meeting focused on overcoming obstacles to better serve students by applying concepts like Maslow's hierarchy of needs, developing common strategies, and using the 7 habits of highly effective people. Participants were encouraged to share ideas openly and problem solve scenarios through the lenses of student needs, effective habits, and applicable strategies with the overall goal of fostering collaboration and innovation to support teaching and learning while having fun and focusing on student success.
Este documento proporciona instrucciones sobre la preparación de una máquina de coser y el corte de telas. Explica cómo colocar las tijeras correctamente, cortar siguiendo las líneas del patrón y doblar cada pieza a medida que se corta. También describe cómo distribuir los patrones en el orden correcto sobre la tela y fijarlos con alfileres, y aumentar las distancias si los patrones no incluyen costuras. Por último, explica la disposición de las telas, incluida la orientación de los hilos longitudinales y trans
Este documento resume los conceptos básicos del flujo en canales abiertos, incluyendo la clasificación de canales, las secciones transversales comunes, los tipos de flujo, y las fórmulas utilizadas para calcular la velocidad y caudal. Define términos como calado, área mojada, radio hidráulico, y número de Froude. También explica conceptos como flujo uniforme, flujo variado, resalto hidráulico, y cómo se calculan las pérdidas de carga.
Presentation: Digital Financial Wellness: The Future of Money
Presented by: Mohamed Khalil, Head of Product, Data & Marketing, Moven
Mobile technology coupled with data and behavioral sciences now allow for highly personalized, real time interactions that alter consumer behaviors. This talk will examine how the retail financial services model must adapt to survive this digital disruption.
http://www.bdionline.com/
The document summarizes the author's pilgrimage to religious sites in South Karnataka associated with Adi Sankara Bhagavatpada, a famous 8th century Indian philosopher. They traveled from their home in Thrissur, Kerala to destinations like Udupi, Kollur Mookambika Temple, Sringeri Sarada Mutt, and Mangalore. These places were important to Sankara and blessed by his presence. The author describes their multi-day journey, visits to temples, and experiences at each location on the trail of Sankara.
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This document summarizes a study on developing a novel protein interaction platform to study neurodegeneration. The study cultured neuronal stem cells to form neurospheres, which were used as a model system. Lentiviral transfection was optimized to introduce target genes stably into the neurospheres. The goal was to analyze protein interactions of Alzheimer's disease proteins by co-immunoprecipitation of neurosphere cultures expressing tagged target proteins. This model aims to further the understanding of molecular mechanisms in neurodegenerative disorders like Alzheimer's disease.
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Caffeine Blocks HIV-1 Tat-Induced Amyloid Beta Production and Tau Phosphoryla...Mahmoud Lotfy Soliman
Caffeine may help reduce Alzheimer's disease pathology associated with HIV infection by blocking the effects of the HIV protein Tat. The study found that:
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3) Caffeine on its own decreased amyloid-beta levels and may help protect against Alzheimer's disease progression in people living with HIV through its effects on endolysosome function and Tat protein activity.
Systemic Lupus Erythematosus (SLE) is an autoimmune disease characterized by autoantibodies that can cause injury to multiple organs. The exact cause is unknown but involves genetic, immunological, and environmental factors. Key aspects include the presence of various nuclear and cytoplasmic autoantibodies, defective B and T cell tolerance, and exposure to environmental triggers. The autoantibodies can cause direct tissue damage or form immune complexes that activate complement pathways. This leads to manifestations in various organs like the kidneys, blood vessels, skin, and joints. Renal involvement is a major cause of morbidity and mortality in SLE patients.
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HIV-1 Nef Gene’s Effect on the Production of Beta Amyloid
Parth Patel
Life Sciences
Woodstock High School
2010 Towne Lake Hills South Dr.
Woodstock, GA 30189
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Synopsis
HIV-1 Nef Gene’s Effect on the Production of Amyloid
HIV/AIDS is a pandemic that kills millions of people. HIV-1 may induce Alzheimer-like
dementia. Alzheimer’s destroys the lives of millions people through the slow decay of brain cells
causing memory loss. In Alzheimer’s the protein beta amyloid is responsible for producing
plaques, which lead to inflammation and cell death. Studies have shown beta amyloid peptides
secreted from cells in association with exosomes. Exosomal proteins were also found to
accumulate in plaques, suggesting a role for exosomes in the pathogenesis of Alzheimer’s. HIV
Nef protein has shown to be secreted in exosomes and enhances the production of exosomes. The
hypothesis is “If beta amyloid is expressed in association with Nef exosomes, and I transfect
NS20Y cells with the nef gene, collect that supernatant, and place it on un-transfected NS20Y
cells, then the production of beta amyloid by the treated cells should be enhanced”. Nef-GFP was
transfected into NS20Y cells to produce Nef exosomes. Medium, containing exosomes, was
collected and placed onto untreated NS20Y cells. ELISA and Western blot measured beta
amyloid production from treated cells. Results show that Nef exosomes (applied exogenously)
enhance the secretion of beta amyloid peptides.
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Table of Contents
Title Page Page 1
Synopsis Page 2
Table of Contents Page 3
Introduction Page 4
Purpose, Procedure, and Materials Page 6
Results Page 8
Conclusion Page 9
Literature Cited Page 12
Appendices Page 13
Acknowledgments Page 17
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Introduction
HIV/AIDS infection is a worldwide epidemic and according to
According to UNAIDS
there were 1.7 million AIDS deaths in 2011, which is actually down from 2.3 million in 2005.
This shows that treatments for HIV/AIDS are improving and more lives are being saved.
However, one complication of HIV-1 infection known as HIV Associated Dementia (HAD),
which is a form of dementia similar to that found in Alzheimer’s patients, has been increasing in
incidence even with modern drug treatments according to an HIV associated neurological
incidence study.
The interesting thing about HAD which was sought to exploit through the research was
its similarities to Alzheimer’s disease. Alzheimer’s is another worldwide epidemic that is
destroying the lives of millions, and in Alzheimer’s disease, there is a protein called beta
amyloid which is responsible for producing plaques (See Figure 1.0) on brain cells, which leads
to inflammation and ultimately cell death resulting in effects like memory loss.
Figure 1.0 – Beta Amyloid Plaque Formation
("Beta Amyloid Formation")
Previous studies have shown that beta amyloid peptides can be secreted from cells in
association with exosomes according to study done specifically on the association between beta
amyloid and exosome excretion; exosomes are small vesicles that have the ability to carry viral
and cellular components (See Figures 2.0).
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Figure 2.0 – Exosome Basics
(Théry)
Subsequently, exosomal proteins were also found to accumulate in the plaques of
Alzheimer’s patient brains, which could be suggesting a role for exosomes in the pathogenesis of
Alzheimer’s disease. Interestingly, HIV Nef, a product of the nef gene (one of six accessory
genes), has shown to be secreted in exosomes and enhances the production of exosomes, so if
more exosomes are associated with the enhanced produciton of beta amyloid, then in turn the nef
gene could be possibly enhancing the secretion of beta amyloid.
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Purpose, Procedure, and Materials
The purpose of this experiment was to study the pathogenesis of HAD, and the possible
implications of the pathogenesis Alzheimer’s disease being related to that of HAD by testing to
see if Nef exosomes are taken up brain cells, then it would enhance the secretion of beta
amyloid. Simultaneously, the purpose was to observe whether or not beta amyloid is expressed
along with exosomes from the nef gene. This experiment is the first of many because its main
purpose was to determine whether or not the nef gene enhances the production of beta amyloid.
Furthermore, the experiment was conducted using a mouse brain cell line called NS20Y, and the
brain cells were transfected with the nef gene. This means the results of this experiment may not
be true for humans, beside the fact that mouse brain cells were used, because HIV has only been
known to infect blood cells. However, if this hypothesis proves to be true, then further studies
may be conducted using human brain and blood cells, eventually leading to method of blocking
the pathogenesis of HAD.
First, the NS20Y cells were split; the media consisted of 10% FBS, DMEM, 1% Glutamax,
1% Antibiotic, and Antymycotic. Then, Nef-GFP was transfected into NS20Y cells, using the
Lipofectamine 2000 Kit, to produce Nef exosomes. A mock and Nef-GFP flasks were created.
The mock was given 20 uL of Lipofectamine, and the Nef-GFP flask was given 10 ug (20.78 uL)
of Nef-GFP (Nef is tagged with GFP in order to observe transfects rate) solution Next, the cells
were split into a 6-well plate and triplicated with .5mL, 1mL, and 1.5mL of Nef-GFP and Mock
supernatants (See Figure 3.0).
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Figure 3.0- Samples
(Created by Researcher)
The resulting conditioned medium was then micro-centrifuged at 16.1 rpm for 10
minutes. Then, the conditioned medium, containing the Nef exosomes, was collected and placed
onto untreated NS20Y cells, and a fluorescent microscope was used to observe whether or not
the brain cells took up the exosomes in the supernatant. Lastly, beta amyloid production from the
treated cells was measured by ELISA using the Mouse/Rat Beta Amyloid (1-40) High Specific
Assay, and beta amyloid precursor protein was measured by Western blot techniques with gels
that were developed for two, eight, and sixty minutes. Next summer, the plans are to continue
research and repeat the same experiment on a different mouse brain cell line, ATT20wt, and on
human brain cells.
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Results
The .5mL NS20Y control (mock) treated cells showed no green fluorescent glow, but the
.5mL Nef-GFP treated cells did express a green fluorescent glow as a result of the nef gene being
tagged with GFP, which indicated that an uptake of vesicles (exosomes) occurred. Additionally,
the 1mL NS20Y control treated cells showed no green fluorescent glow while the 1ml Nef-GFP
treated cells showed a greater uptake of exosomes occurred. However, the 1.5mL NS20Y Nef-
GFP treated cells showed a decrease in the uptake of exosomes while the NS20Y control treated
cells showed no green fluorescent glow, which could be suggesting that there is a limit to how
many exosomes can be taken up.
The ELISA results for the Mouse/Rat Beta Amyloid (1-40) High Specific Assay showed
the amount of beta amyloid found in the supernatant of the mock cells and the nef-transfected
cells. The original or stock solution of the mock-transfected cells secreted 211pg/mL, and the
nef-transfected cells secreted 641pg/mL. The .5mL solution of mock-transfected cells secreted
179pg/mL while the nef-transfected cells secreted 49pg/mL, which was the least. The second
most secreted was 177pg/mL by the 1.5mL mock-transfected cells and 235 by the 1.5ml nef-
transfected cells. The greatest amount of beta amyloid secreted was 263pg/mL by the 1mL
mock-transfected cells and 300pg/Ml by the 1mL nef-transfected cells (See Figure 4.0).
Figure 4.0 – Mouse/Rat Beta Amyloid (1-40) High Specific Assay Results
(Created by Researcher)
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The sixty minute developed western blot showed the clearest results and was used to
observe the amount of beta amyloid precursor protein found within the lysate mock and nef-
transfected cells. The western blot results showed that the 1mL nef-transfect cells had the
greatest trace or amount of beta amyloid precursor protein; however, other specific results are
not presentable because the data is too difficult to examine (See Figure 5.0).
Figure 5.0 – Western Blot Results
(Created by Researcher)
Conclusion
The experiment has supported the hypothesis that the uptake of Nef exosomes by brain
cells would enhance the secretion of beta amyloid.
Nef-GFP vesicles produced from transfected
NS20Y cells appear to be able to change the amount of beta amyloid secreted when applied to
un-transfected cells, and therefore, it can be concluded that Nef may play a role in HIV
Associated Dementia by modulating the secretion of beta amyloid. However, in the ELISA and
western blot results there may have been a mix up with the .5mL supernatant solution for the
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mock-transfacted and nef-transfected cells. However, this does not the affect the conclusion that
the uptake of exosomes by brain cells enhances the production of beta amyloid. As a result,
further conclusions cannot be drawn until more trials are conducted and experimentation is
conducted in consideration of the human body’s immune system response.
In the continuation of this project, there are various paths that may be taken. This
summer, the plan is to conduct this same experiment on human brain cells and then with human
blood cells because HIV cannot directly infect brain cells because of the blood barrier, but it is
possible that HIV hijacks a pathway to the brain and sends exosomes to infect the brain cells.
This seems logical and plausible because the vesicles would be able to bypass the brain’s blood
barrier. As a result, the Nef exosomes would be mimicking how HIV attacks T-cells (See Figure
6.0).
Figure 6.0 – HIV T-cell Infection
(Created by Dr. Andrea Raymond)
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However, it is extremely difficult to obtain live human brain cells, but once this obstacle
is overcome, blocking the Nef enhancement of beta amyloid production could represent a new
treatment for AIDS dementia.
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Literature Cited
United Nations. UNAIDS. 2012 UNAIDS Report on the Global AIDS Epidemic. N.p.:
Joint United Nations Programme on HIV/AIDS (UNAIDS), 2012. Print.
Sacktor, N. HIV-associated neurologic disease incidence changes: Multicenter
AIDS Cohort Study, 1990–1998. Comp. R. H. Lyles et al. N.p.: n.p., 2001.
Print.
Planck, Max. Alzheimer's disease beta-amyloid peptides are released in
association with exosomes. Comp. L. Rajendran et al. N.p.: n.p., 2006.
Print.
"Beta Amyloid Formation." Elements4Health. Elements4Health, 2014. Web. 17 June
2013. <http://www.elements4health.com/images/stories/
amyloid-plaque-formation.jpg>.
Théry, Clotilde. "Exosome Basics." TheScientist. The Scientist, 1 July 2011.
Web. 12 June 2013. <http://www.the-scientist.com/?articles.view/
articleNo/30791/title/Exosome-Basics/>.
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Figures 7.1, 8.1, and 9.1 are not the pictures of the NS20Y cells from this experiment
because the pictures from this experiment were lost. However, these pictures are accurate
measurements in the ratio of the amount NS20Y cells that took up the exosomes and displayed
green fluorescent glow.
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Acknowledgments
This experiment and research was conducted at Morehouse School of Medicine in
Biosafety Hazard Level 2 laboratory for six weeks throughout June and July of 2013.
Dr. Michael D. Powell as the primary investigator expertly taught the biological part of
the experiment and presented various types of literature consisting of background information
regarding the research. This tremendously helped in grasping the concepts firmly and quickly.
Dr. Powell also provided assistance in formulating the hypothesis and drawing conclusions,
making sure none of the conclusions were invalid or extrapolation. Dr. Powell was a great
mentor, teacher, and friend throughout the duration of the research. He is also responsible for
retrieving financial sponsorship and providing the brain cells, Lipofectamine Kit, media, and
Mouse/Rat Beta Amyloid (1-40) High Specific Assay Kit.
Mrs. Mahfuz Khan, the research assistant, assisted in the experimental design,
demonstrated how special equipment, such as the NanoDrop and fluorescent microscope, were to
be used, and taught various lab techniques that were vital to gathering the most accurate data.
She also demonstrated how to conduct ELISA and western blot tests.
Lastly, the Morehouse School of Medicine was generous enough to allow access to its
equipment and providing this research experience through the Vivien Thomas Summer Research
Program.