Osteoprotegerin and commensal bacteria
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OPG expression in intestinal epithelial cells appears to be suppressed when exposed to commensal bacteria. OPG binds to both live and killed bacteria and this bound OPG can be released through detergent treatment. Treatment of Caco 2 cell supernatants with detergent does not increase detectable OPG levels by ELISA, indicating ELISA detects the true OPG concentration.
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Kupffer Cells Mediate Leptin-Induced Liver Fibrosis.
GASTROENTEROLOGY 2009;137:713–723
JIANHUA WANG,* ISABELLE LECLERCQ,‡ JOANNE M. BRYMORA,* NING XU,* MEHDI RAMEZANI–MOGHADAM,* ROSLYN M. LONDON,* DAVID BRIGSTOCK,§ and JACOB GEORGE*
*Storr Liver Unit, Westmead Millennium Institute, University of Sydney and Westmead Hospital, Westmead, Australia; ‡Laboratory of Gastroenterology, Faculty of
Medicine, Université Catholique de Louvain, Brussels, Belgium; and §Center for Cell and Vascular Biology, Children’s Research Institute, Columbus, Ohio
瘦素(Leptin)是一由脂肪細胞(Adipocyte)所分泌之荷爾蒙，是調控體重及新陳代謝之重要因子。過去研究發現病態肥胖(Obese)、脂肪肝(Nonalcoholic steatohepatitis)及酒精性肝炎(Alcoholic liver disease)等病患之血液循環中，Leptin量有明顯增加。而近期研究報告指出leptin具有促進肝臟纖維化(Liver fibrosis)之能力，當中分子機理並未明確。
在肝纖維化過程中，肝臟星狀細胞(HSC)會被活化增生及促進胞外基質(ECM)產生，而鄰近之Kupffer細胞(KC)則已知可透過促發炎因子(Proinflammatory factor)和促纖維化因子(Profibrogenic factors)例如TGF-β1和ROS影響HSC表現。雖然HSC是肝纖維化過程中重要角色，前人研究卻發現leptin似對HSC無任何調控作用。故本篇作者針對Leptin是否透過間接作用於HSC鄰近之KC，刺激其產生促纖維化因子，以活化HSC。
為探討leptin直接或間接影響HSC之分子機理，本篇作者透過RT-PCR、Immunoblot等分子生物學方法，分別測定leptin刺激後HSC及KC中Collagen I、TIMP1等促纖維化因子基因及蛋白表現，發現leptin雖可促使HSC增生，但對其纖維化能力之影響甚微。而leptin可刺激KC中TGF-β1及CTGF/CCN2等肝纖維化中重要之cytokines表現。另發現Leptin-treated KC-conditioned培養液可刺激HSC增生及增加其中Collagen I、TIMP1等表現，得出了leptin是透過刺激KC來活化HSC之推論。作者亦於後續實驗中，透過磷酸化測定、EMSA等方法探討leptin訊號傳遞作用，發現leptin可活化KC中STAT3、ERK1/2、AKT等路徑，及下游因子AP-1、NF-κB，而此兩種蛋白具有增強TGF-β1及CTGF/CCN2基因表現之能力。
Evgeny nikolaev proteomics of body liquids as a source for potential methods ...
The document discusses using mass spectrometry to analyze body fluids like urine, saliva, and exhaled breath condensate for medical diagnostics and biomarker discovery. It describes creating databases of accurate mass tags and retention times from mass spec analyses of peptides and proteins in body fluids to enable fast identification. Biomarkers found for diseases like COPD, pneumonia and changes after lung transplantation are mentioned.
Fall project
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.
Crizotinib a8081001 asco 2010 slides
This document summarizes clinical trial results of the oral ALK inhibitor crizotinib (PF-02341066) in patients with ALK-positive non-small cell lung cancer. The trial showed that crizotinib had significant clinical activity in ALK-positive NSCLC patients, with an objective response rate of 57% and disease control rate of 87%. The median progression-free survival was 10 months for these patients. In contrast, patients with ALK-negative NSCLC did not respond to crizotinib. These results provide evidence that crizotinib is an effective targeted therapy for patients with ALK-positive NSCLC.
Ablooglu, AJ (2010) Development
Integrin V can form heterodimers with several subunits to mediate cell-cell and cell-extracellular matrix interactions. During zebrafish gastrulation, V is expressed maternally and zygotically. Here, we used a morpholino-mediated V knockdown strategy to study V function. Although V morphants displayed vascular defects, they also exhibited left-right body asymmetry defects affecting multiple visceral organs. This was preceded by mislocalization of dorsal forerunner cells (DFCs) and malformation of the Kupffer’s vesicle (KV) laterality organ
Faseb poster2007b
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
An Understanding Of Bacterial Transformation By Plasmid Dna
Bacterial plasmids are small, circular DNA molecules within bacteria that are separate from the bacterial chromosome. Plasmids can contain genes that provide bacteria with useful traits like antibiotic resistance. During genetic transformation, the plasmid is introduced to recipient bacteria where it can be replicated independently of the bacterial chromosome. The foreign DNA from the plasmid is then expressed in the recipient bacteria, altering its genotype and phenotype. This allows bacteria to horizontally acquire new genes from plasmids and gain traits like antibiotic resistance without direct contact between bacterial cells.
Structural Studies of Human GBE1 and Relevance to APBD
The document summarizes Wyatt Yue's work studying the structure of human GBE1 and its relevance to Adult Polyglucosan Body Disease (APBD). Key points:
1. Wyatt determined the crystal structures of human GBE1 in its apo form and bound to acarbose and Glc7, to understand its shape and function.
2. Mutations in GBE1 cause APBD. Wyatt purified several GBE1 mutants to characterize, including Y329S, and determined the Y329 site is disrupted in the mutant structure.
3. Wyatt collaborates with APBD patient groups like APBDRF to translate structural findings into potential pharmacological chaperone treatments for
11
Mutations that reduce outer membrane permeability in Escherichia coli lead to increased tolerance of the bacterium to the antimicrobial lactoperoxidase enzyme system. The study identified two E. coli mutants with increased tolerance to lactoperoxidase due to mutations in genes involved in lipopolysaccharide synthesis. These mutants had reduced amounts of porins in their outer membrane, indicating decreased outer membrane permeability. Knockout mutants of specific porin genes also showed increased tolerance, suggesting the antimicrobial activity of lactoperoxidase relies on uptake of its oxidized substrates through porins.
The Beta Amyloid Cleaving Enzymes: From Drug Discovery to Evolution and Back
Slides presented at the University of Edinburgh in Jan 2014, based on our paper "A tale of two drug targets: the evolutionary history of BACE1 and BACE2" http://www.ncbi.nlm.nih.gov/pubmed/24381583
The beta amyloid (APP) cleaving enzyme (BACE1) has been a drug target for Alzheimer's Disease (AD) since 1999 with lead inhibitors now entering clinical trials. In 2011, the paralog, BACE2, became a new target for type II diabetes (T2DM) having been identified as a TMEM27 secretase regulating pancreatic β cell function. However, the normal roles of both enzymes are unclear. This study outlines their evolutionary history and new opportunities for functional genomics. We identified 30 homologs (UrBACEs) in basal phyla including Placozoans, Cnidarians, Choanoflagellates, Porifera, Echinoderms, Annelids, Mollusks and Ascidians (but not Ecdysozoans). UrBACEs are predominantly single copy, show 35–45% protein sequence identity with mammalian BACE1, are ~100 residues longer than cathepsin paralogs with an aspartyl protease domain flanked by a signal peptide and a C-terminal transmembrane domain. While multiple paralogs in Trichoplax and Monosiga pre-date the nervous system, duplication of the UrBACE in fish gave rise to BACE1 and BACE2 in the vertebrate lineage. The latter evolved more rapidly as the former maintained the emergent neuronal role. In mammals, Ka/Ks for BACE2 is higher than BACE1 but low ratios for both suggest purifying selection. The 5' exons show higher Ka/Ks than the catalytic section. Model organism genomes show the absence of certain BACE human substrates when the UrBACE is present. Experiments could thus reveal undiscovered substrates and roles. The human protease double-target status means that evolutionary trajectories and functional shifts associated with different substrates will have implications for the development of clinical candidates for both AD and T2DM. A rational basis for inhibition specificity ratios and assessing target-related side effects will be facilitated by a more complete picture of BACE1 and BACE2 functions informed by their evolutionary context.
Presentation-Moscow2005
PAR3 is expressed on mouse lymphocytes and myeloid cells, particularly B lymphocytes. The PAR3 receptor on B lymphocytes is functionally active and involved in regulating lymphocyte proliferation and activation in response to thrombin. PAR3 may represent an important link between the coagulation and immune systems. A monoclonal antibody (mAb 8E8) was characterized that specifically binds to the cleavage site of mouse PAR3. This antibody blocked thrombin-induced platelet aggregation and calcium signaling in platelets and splenocytes. It also inhibited proliferation of a hybridoma cell line in response to thrombin or PAR3 peptides but not PAR4 peptides. The antibody further suppressed activation of splenocytes induced by anti-CD3/anti-CD40 stimulation
Evaluating Hepatocyte-derived mESCs
This document summarizes a study evaluating the efficacy of differentiating mouse embryonic stem cells (mESCs) into hepatocyte-like cells. The researchers found that cell cycle synchronization of mESCs prior to differentiation led to decreased expression of the pluripotency marker Oct4 and increased expression of endoderm and hepatic progenitor markers, indicating improved differentiation. Assays of stem cell-derived hepatocyte-like cells showed production of the liver proteins albumin and urea, though urea levels decreased over time, suggesting further optimization is needed to generate fully functional hepatocytes from mESCs. The researchers concluded cell cycle synchronization enhanced differentiation but that identifying additional progenitor factors could improve hepatic function of the derived cells.
Fernandez 111309
Presentation of my work on dendritic cell lineage-committed mouse embryonic stem cells for transplantation tolerance
Hofstetter PON1 meeting 2015 KOmice corrected
- Serum carboxylesterase knockout (sCaE KO) mice lack an enzyme that provides increased protection against certain organophosphorous nerve agents in mice and rats compared to primates.
- Analysis found that sCaE KO mice are physiologically similar to wild-type mice except for the absence of the carboxylesterase enzyme in their blood.
- sCaE KO mice were found to have LD50 values for G-series nerve agents that were 20-40% of those for wild-type mice, indicating they are a more relevant model for predicting human responses compared to other small animal models.
- Injection of bioscavenger enzymes in sCaE KO mice protected them against
CRYOPRESERVATION.pptx
This study investigated the effects of vitrification on morphology and mRNA expression of apoptotic genes in sheep immature cumulus oocyte complexes. Immature oocytes were collected from sheep ovaries and divided into groups for cryopreservation using different cryoprotectants. The oocytes were evaluated for morphology before and after vitrification and thawing. Real-time PCR was used to analyze expression of apoptotic genes BAD, BAK, BAX, BID, BOK, BCL2A1, BCL2, MCL1 in fresh and vitrified oocytes. Results showed varying recovery and damage rates among oocytes cryopreserved with different cryoprotectants. Morphological changes included cracked or split zona pell
CRYOPRESERVATION.pptx
This study investigated the effects of vitrification on morphology and mRNA expression of apoptotic genes in sheep immature cumulus oocyte complexes. Immature oocytes were collected from sheep ovaries and divided into groups for cryopreservation using different cryoprotectants. The oocytes were evaluated for morphology before and after vitrification and thawing. Real-time PCR was used to analyze expression of apoptotic genes BAD, BAK, BAX, BID, BOK, BCL2A1, BCL2, MCL1 in fresh and vitrified oocytes. Results showed varying recovery and damage rates among oocytes cryopreserved with different cryoprotectants. Morphological changes included cracked or split zona pell
Apoptosis of lymphocytes in SLE
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.
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Deciphering the role of osteoprotegerin in inflammatory bowel diseases
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Evgeny nikolaev proteomics of body liquids as a source for potential methods ...
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An Understanding Of Bacterial Transformation By Plasmid Dna
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The Beta Amyloid Cleaving Enzymes: From Drug Discovery to Evolution and Back
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The use of Nauplii and metanauplii artemia in aquaculture (brine shrimp).pptx
Although Artemia has been known to man for centuries, its use as a food for the culture of larval organisms apparently began only in the 1930s, when several investigators found that it made an excellent food for newly hatched fish larvae (Litvinenko et al., 2023). As aquaculture developed in the 1960s and ‘70s, the use of Artemia also became more widespread, due both to its convenience and to its nutritional value for larval organisms (Arenas-Pardo et al., 2024). The fact that Artemia dormant cysts can be stored for long periods in cans, and then used as an off-the-shelf food requiring only 24 h of incubation makes them the most convenient, least labor-intensive, live food available for aquaculture (Sorgeloos & Roubach, 2021). The nutritional value of Artemia, especially for marine organisms, is not constant, but varies both geographically and temporally. During the last decade, however, both the causes of Artemia nutritional variability and methods to improve poorquality Artemia have been identified (Loufi et al., 2024).
Brine shrimp (Artemia spp.) are used in marine aquaculture worldwide. Annually, more than 2,000 metric tons of dry cysts are used for cultivation of fish, crustacean, and shellfish larva. Brine shrimp are important to aquaculture because newly hatched brine shrimp nauplii (larvae) provide a food source for many fish fry (Mozanzadeh et al., 2021). Culture and harvesting of brine shrimp eggs represents another aspect of the aquaculture industry. Nauplii and metanauplii of Artemia, commonly known as brine shrimp, play a crucial role in aquaculture due to their nutritional value and suitability as live feed for many aquatic species, particularly in larval stages (Sorgeloos & Roubach, 2021).
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Orion Air Quality Monitoring Systems - CWS
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
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如何办理(uvic毕业证书)维多利亚大学毕业证本科学位证书原版一模一样
原版纸张【微信：741003700 】【(uvic毕业证书)维多利亚大学毕业证】【微信：741003700 】学位证，留信认证（真实可查，永久存档）offer、雅思、外壳等材料/诚信可靠,可直接看成品样本，帮您解决无法毕业带来的各种难题！外壳，原版制作，诚信可靠，可直接看成品样本。行业标杆！精益求精，诚心合作，真诚制作！多年品质 ,按需精细制作，24小时接单,全套进口原装设备。十五年致力于帮助留学生解决难题，包您满意。
本公司拥有海外各大学样板无数，能完美还原海外各大学 Bachelor Diploma degree, Master Degree Diploma
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留信网认证的作用:
1:该专业认证可证明留学生真实身份
2:同时对留学生所学专业登记给予评定
3:国家专业人才认证中心颁发入库证书
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6:个人职称评审加20分
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Nucleophilic Addition of carbonyl compounds.pptx
Nucleophilic addition is the most important reaction of carbonyls. Not just aldehydes and ketones, but also carboxylic acid derivatives in general.
Carbonyls undergo addition reactions with a large range of nucleophiles.
Comparing the relative basicity of the nucleophile and the product is extremely helpful in determining how reversible the addition reaction is. Reactions with Grignards and hydrides are irreversible. Reactions with weak bases like halides and carboxylates generally don’t happen.
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Deep Software Variability and Frictionless Reproducibility
Deep Software Variability and Frictionless ReproducibilityUniversity of Rennes, INSA Rennes, Inria/IRISA, CNRS
The ability to recreate computational results with minimal effort and actionable metrics provides a solid foundation for scientific research and software development. When people can replicate an analysis at the touch of a button using open-source software, open data, and methods to assess and compare proposals, it significantly eases verification of results, engagement with a diverse range of contributors, and progress. However, we have yet to fully achieve this; there are still many sociotechnical frictions.
Inspired by David Donoho's vision, this talk aims to revisit the three crucial pillars of frictionless reproducibility (data sharing, code sharing, and competitive challenges) with the perspective of deep software variability.
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I will then present some software engineering and AI techniques that can support the strategic exploration of variability spaces. These include the use of abstractions and models (e.g., feature models), sampling strategies (e.g., uniform, random), cost-effective measurements (e.g., incremental build of software configurations), and dimensionality reduction methods (e.g., transfer learning, feature selection, software debloating).
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EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...
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The use of Nauplii and metanauplii artemia in aquaculture (brine shrimp).pptx
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Osteoprotegerin and commensal bacteria
- 1. Recent developments in understanding the role of osteoprotegerin in colonic intestinal epithelium at Sylvester lab Raghunath Ramanarasimhaiah
- 2. Which evolved first? Gut or bone! http://answers.mheducation.com/sciences/life-science/biology/animal-kingdom
- 3. Expression of OPG in different cell types van Agtmaal EL, et al. The shear stress-induced transcription factor KLF2 affects dynamics and angiopoietin-2 content of Weibel-Palade bodies. PLoS One. 2012;7(6):e38399. Chollet ME, et al. Evidence of a colocalisation of osteoprotegerin (OPG) with von Willebrand factor (VWF) in platelets and megakaryocytes alpha granules. Studies from normal and grey platelets. Br J Haematol. 2010 Mar;148(5):805-7. Sylvester lab MG63 osteoblastic cells OPG DAPI Merge Caco 2 Intestinal epithelial cells
- 4. Expression of OPG in different tissues Carda C, et al. Osteoprotegerin (OPG) and RANKL expression and distribution in developing human craniomandibular joint. Tissue Cell. 2005 Jun;37(3):247-55. Ueland T, et al. Dysregulated osteoprotegerin/RANK ligand/RANK axis in clinical and experimental heart failure. Circulation. 2005 May 17;111(19):2461-8. Sylvester lab
- 5. Questions • Does treatment with a detergent increase detectable concentration of osteoprotegerin (OPG) in spent Caco 2 media? • Does exposure to commensal bacteria alter OPG expression in Caco 2 cells? • Does OPG bind to bacteria? If yes, can it be released from the bacteria? • Expression of OPG in appendix?
- 6. Question • Does treatment with a detergent increase detectable concentration of OPG in spent Caco 2 media?
- 7. Experimental procedure Spin down Collect supernatant ELISAIL1β Media only Add 1% Tween 20 Incubate on ice for 30 M 12 well culture plate supporting Caco 2 Over night 370C Spin down Collect supernatant Add 1% Tween 20 Incubate on ice for 30 M ELISA
- 8. Detergent treatment of Caco 2 supernatant does not increase the detectable OPG by ELISA 3000 6000 9000 12000 15000 1% Tween 20 IL1β - - ++ + -
- 9. Cell culture inserts • Seed and grow Caco 2 cells as a monolayer • After achieving confluence, allow 21 days for polarization • Check TEER (transepithelial electrical resistance) Apical Cell monolayer Baso-lateral
- 10. Questions • Does exposure to commensal bacteria (ATCC 25922) alter OPG expression in Caco 2 cells?
- 11. Experimental procedure Live E.coli Spindown Collect supernatant ELISA Apical Baso lateral Over night 370C
- 12. OPG expression appears suppressed on exposure to commensal bacteria A B A B A B A B A B A B 106 105 104 102 EC Sup Media 2000 4000 6000 8000 10000 No. of EC/insert A: Apical B: Basolateral EC: Escherichia coli EC Sup: Filtered spent medium of EC pg/mLOPG
- 13. Questions • Does OPG bind to bacteria? If yes, can it be released from the bacteria?
- 14. Experimental procedure Spindown Collect supernatant ELISAOn ice 30 M 0.05% Tween 20 Spindown Collect supernatant ELISAOn ice 30 M ELISA Spindown Live E.coli Killed E.coli Over night 370C TEER
- 15. Live Escherichia coli + Live E.coli 106 104 102 EC sup EC med Con med 2000 4000 6000 - +- +- +- +- +- EC sup: E.coli spent media EC med: Fresh LB broth Con med: Caco 2 media Tween 20 (0.05%)
- 16. Killed Escherichia coli Tween 20 (0.05%) Killed E.coli 106 104 102 EC sup EC med Con med 2000 4000 6000 +- +- +- +- +- +- EC sup: E.coli spent media EC med: Fresh LB broth Con med: Caco 2 media
- 17. Baso-lateral compartment L K + - + - + -E.coli 106 104 102 EC sup EC med Con med 2000 4000 6000 L K L K L: Live K: Killed EC sup: E.coli spent media EC med: Fresh LB broth Con med: Caco 2 media
- 18. Conclusions • ELISA detects the true concentration of OPG in Caco 2 supernatants and is not altered by detergent (0.05% Tween 20) • Osteoprotegerin (OPG) binds to commensal bacteria and the bound OPG can be released by detergent treatment