This work was supported by a Howard Hughes Medical Institute Education Grant and an internal grant from the Saint Joseph's University chapter of Sigma Xi. We would also like to thank the three anonymous reviewers who helped strengthen the protocol and interpretations.

<ol>
	<li>Huntingford, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relationship+between+anti-predator+behaviour+and+aggression+among+conspecifics+in+the+three-spined+stickleback,+Gasterosteus+aculeatus.">The relationship between anti-predator behaviour and aggression among conspecifics in the three-spined stickleback, Gasterosteus aculeatus.</a> Anim Behav. 24, 245-260 (1976).</li><li>R&#233;ale, D., Reader, S. M., Sol, D., McDougall, P. T., Dingemanse, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrating+animal+temperament+within+ecology+and+evolution.">Integrating animal temperament within ecology and evolution.</a> Biol Rev. 82 (2), 291-318 (2007).</li><li>Sih, A., Bell, A., Johnson, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+syndromes:+an+ecological+and+evolutionary+overview.">Behavioral syndromes: an ecological and evolutionary overview.</a> Trends Ecol Evol. 19 (7), 372-378 (2004).</li><li>Conrad, J. L., Weinersmith, K. L., Brodin, T., Saltz, J. B., Sih, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioural+syndromes+in+fishes:+a+review+with+implications+for+ecology+and+fisheries+management.">Behavioural syndromes in fishes: a review with implications for ecology and fisheries management.</a> J Fish Biol. 78 (2), 395-435 (2011).</li><li>Wolf, M., Weissing, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+personalities:+consequences+for+ecology+and+evolution.">Animal personalities: consequences for ecology and evolution.</a> Trends Ecol Evol. 27 (8), 452-461 (2012).</li><li>Langheinrich, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zebrafish:+a+new+model+on+the+pharmaceutical+catwalk.">Zebrafish: a new model on the pharmaceutical catwalk.</a> BioEssays. 25 (9), 904-912 (2003).</li><li>Dai, Y. -. J., Jia, Y. -. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zebrafish+as+a+model+system+to+study+toxicology:+Zebrafish+toxicology+monitoring.">Zebrafish as a model system to study toxicology: Zebrafish toxicology monitoring.</a> Environ Toxicol Chem. 33 (1), 11-17 (2014).</li><li>Norton, W., Bally-Cuif, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adult+zebrafish+as+a+model+organism+for+behavioural+genetics.">Adult zebrafish as a model organism for behavioural genetics.</a> Neuroscience. 11 (1), 90 (2010).</li><li>Gerlai, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zebra+fish:+an+uncharted+behavior+genetic+model.">Zebra fish: an uncharted behavior genetic model.</a> Behav Genet. 33 (5), 461-468 (2003).</li><li>Dzieweczynski, T. L., Campbell, B. A., Marks, J. M., Logan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+exposure+to+17&#945;-ethinylestradiol+alters+boldness+behavioral+syndrome+in+female+Siamese+fighting+fish.">Acute exposure to 17&#945;-ethinylestradiol alters boldness behavioral syndrome in female Siamese fighting fish.</a> Horm Behav. 66 (4), 577-584 (2014).</li><li>Mikl&#242;si, &. #. 1. 9. 3. ;., Andrew, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Zebrafish+as+a+Model+for+Behavioral+Studies.">The Zebrafish as a Model for Behavioral Studies.</a> Zebrafish. 3 (2), 227-234 (2006).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Zebrafish protocols for neurobehavioral research. , (2012).</li><li>Moretz, J. A., Martins, E. P., Robison, B. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+syndromes+and+the+evolution+of+correlated+behavior+in+zebrafish.">Behavioral syndromes and the evolution of correlated behavior in zebrafish.</a> Behav Ecol. 18 (3), 556-562 (2007).</li><li>Way, G. P., Kiesel, A. L., Ruhl, N., Snekser, J. L., McRobert, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sex+differences+in+a+shoaling-boldness+behavioral+syndrome,+but+no+link+with+aggression.">Sex differences in a shoaling-boldness behavioral syndrome, but no link with aggression.</a> Behav Process. 113, 7-12 (2015).</li><li>Toms, C. N., Echevarria, D. J., Jouandot, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+methodological+review+of+personality-related+studies+in+fish:+Focus+on+the+shy-bold+axis+of+behavior.">A methodological review of personality-related studies in fish: Focus on the shy-bold axis of behavior.</a> J Comp Psychol. 23, 1-25 (2010).</li><li>Rowland, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studying+visual+cues+in+fish+behavior:+A+review+of+ethological+techniques.">Studying visual cues in fish behavior: A review of ethological techniques.</a> Environ Biol Fish. 56 (3), 285-305 (1999).</li><li>Way, G. P., Ruhl, N., Snekser, J. L., Kiesel, A. L., McRobert, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Comparison+of+Methodologies+to+Test+Aggression+in+Zebrafish.">A Comparison of Methodologies to Test Aggression in Zebrafish.</a> Zebrafish. 12 (2), 144-151 (2015).</li><li>Moss, S., Tittaferrante, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interactions+between+aggression,+boldness+and+shoaling+within+a+brood+of+convict+cichlids+(Amatitlania+nigrofasciatus).">Interactions between aggression, boldness and shoaling within a brood of convict cichlids (Amatitlania nigrofasciatus).</a> Behav Process. 121, 63-69 (2015).</li><li>Ruhl, N., Kiesel, A. L., McRobert, S. P., Snekser, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioural+syndromes+and+shoaling:+connections+between+aggression,+boldness+and+social+behaviour+in+three+different+Danios.">Behavioural syndromes and shoaling: connections between aggression, boldness and social behaviour in three different Danios.</a> Behaviour. 149 (10-12), 1155-1175 (2012).</li><li>Larson, E. T., O'Malley, D. M., Melloni, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aggression+and+vasotocin+are+associated+with+dominant-subordinate+relationships+in+zebrafish.">Aggression and vasotocin are associated with dominant-subordinate relationships in zebrafish.</a> Behav Brain Res. 167 (1), 94-102 (2006).</li><li>McRobert, S., Bradner, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+body+coloration+on+shoaling+preferences+in+fish.">The influence of body coloration on shoaling preferences in fish.</a> Anim Behav. 56 (3), 611-615 (1998).</li><li>Ruhl, N., McRobert, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+sex+and+shoal+size+on+shoaling+behaviour+in+Danio+rerio.">The effect of sex and shoal size on shoaling behaviour in Danio rerio.</a> J Fish Biol. 67 (5), 1318-1326 (2005).</li><li>Snekser, J., Ruhl, N., Bauer, K., McRobert, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+sex+and+phenotype+on+shoaling+decisions+in+zebrafish.">The influence of sex and phenotype on shoaling decisions in zebrafish.</a> J Comp Psychol. 23, 70-81 (2010).</li><li>Budaev, S. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+Principal+Components+and+Factor+Analysis+in+Animal+Behaviour+Research:+Caveats+and+Guidelines.">Using Principal Components and Factor Analysis in Animal Behaviour Research: Caveats and Guidelines.</a> Ethology. 116 (5), 472-480 (2010).</li><li>Paciorek, T., McRobert, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Daily+variation+in+the+shoaling+behavior+of+zebrafish+Danio+rerio.">Daily variation in the shoaling behavior of zebrafish Danio rerio.</a> Curr Zool. 58 (1), 129-137 (2012).</li><li>Wright, D., Krause, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repeated+measures+of+shoaling+tendency+in+zebrafish+(Danio+rerio)+and+other+small+teleost+fishes.">Repeated measures of shoaling tendency in zebrafish (Danio rerio) and other small teleost fishes.</a> Nat Protoc. 1 (4), 1828-1831 (2006).</li><li>Cachat, J., Stewart, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+behavioral+and+endocrine+responses+to+novelty+stress+in+adult+zebrafish.">Measuring behavioral and endocrine responses to novelty stress in adult zebrafish.</a> Nat Protoc. 5 (11), 1786-1799 (2010).</li><li>Croft, D. P., Krause, J., Couzin, I. D., Pitcher, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=When+fish+shoals+meet:+outcomes+for+evolution+and+fisheries.">When fish shoals meet: outcomes for evolution and fisheries.</a> Fish Fish. 4, 138-146 (2003).</li><li>Brown, M. B., Forsythe, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Robust+Tests+for+the+Equality+of+Variances.">Robust Tests for the Equality of Variances.</a> J Am Stat Assoc. 69 (346), (1974).</li><li>Kruskal, W. H., Wallis, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+Ranks+in+One-Criterion+Variance+Analysis.">Use of Ranks in One-Criterion Variance Analysis.</a> J Am Stat Assoc. 47 (260), 583-621 (1952).</li><li>Nelder, J. A., Wedderburn, R. W. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generalized+Linear+Models.">Generalized Linear Models.</a> J Roy Stat Soc A Sta. 135, 370-384 (1972).</li><li>Iman, R. L., Conover, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+distribution-free+approach+to+inducing+rank+correlation+among+input+variables.">A distribution-free approach to inducing rank correlation among input variables.</a> Commun Stat Simulat. 11 (3), 311-334 (1982).</li><li>Fleiss, J. L., Cohen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Equivalence+of+Weighted+Kappa+and+the+Intraclass+Correlation+Coefficient+as+Measures+of+Reliability.">The Equivalence of Weighted Kappa and the Intraclass Correlation Coefficient as Measures of Reliability.</a> Educ Psychol Meas. 33 (3), 613-619 (1973).</li><li>Ruhl, N., McRobert, S. P., Currie, W. J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Shoaling+preferences+and+the+effects+of+sex+ratio+on+spawning+and+aggression+in+small+laboratory+populations+of+zebrafish+(Danio+rerio).">Shoaling preferences and the effects of sex ratio on spawning and aggression in small laboratory populations of zebrafish (Danio rerio).</a> Lab Animal. 38 (8), 264-269 (2009).</li><li>Bell, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioural+differences+between+individuals+and+two+populations+of+stickleback+(Gasterosteus+aculeatus).">Behavioural differences between individuals and two populations of stickleback (Gasterosteus aculeatus).</a> J Evolution Biol. 18 (2), 464-473 (2005).</li><li>Wilson, A. D. M., Godin, J. -. G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Boldness+and+behavioral+syndromes+in+the+bluegill+sunfish,+Lepomis+macrochirus.">Boldness and behavioral syndromes in the bluegill sunfish, Lepomis macrochirus.</a> Behav Ecol. 20 (2), 231-237 (2009).</li><li>Brodin, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+syndrome+over+the+boundaries+of+life--carryovers+from+larvae+to+adult+damselfly.">Behavioral syndrome over the boundaries of life--carryovers from larvae to adult damselfly.</a> Behav Ecol. 20 (1), 30-37 (2008).</li><li>Noldus, L. P., Spink, A. J., Tegelenbosch, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EthoVision:+a+versatile+video+tracking+system+for+automation+of+behavioral+experiments.">EthoVision: a versatile video tracking system for automation of behavioral experiments.</a> Behav Res Methods Instrum Comput. 33 (3), 398-414 (2001).</li><li>Cote, J., Fogarty, S., Weinersmith, K., Brodin, T., Sih, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Personality+traits+and+dispersal+tendency+in+the+invasive+mosquitofish+(Gambusia+affinis).">Personality traits and dispersal tendency in the invasive mosquitofish (Gambusia affinis).</a> Proc R Soc B. 277 (1687), 1571-1579 (2010).</li><li>Fraser, D. F., Gilliam, J. F., Daley, M. J., Le, A. N., Skalski, G. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Explaining+leptokurtic+movement+distributions:+intrapopulation+variation+in+boldness+and+exploration.">Explaining leptokurtic movement distributions: intrapopulation variation in boldness and exploration.</a> Am Nat. 158 (2), 124-135 (2001).</li><li>Dingemanse, N. J., Wright, J., Kazem, A. J. N., Thomas, D. K., Hickling, R., Dawnay, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioural+syndromes+differ+predictably+between+12+populations+of+three-spined+stickleback.">Behavioural syndromes differ predictably between 12 populations of three-spined stickleback.</a> J Anim Ecol. 76 (6), 1128-1138 (2007).</li></ol>

The authors (GW and SPM) have no competing financial interests or conflicts of interest.

הפרוטוקול יקבע אם יש עמותות עקביות תעוזה, תוקפנות, והתנהגויות shoaling דג הזברה. אם יש עמותות עקביות באוכלוסייה נתונה בין כל ההתנהגויות האלה, ואז תסמונת התנהגותית היא הווה. על ידי לימוד תסמונת התנהגותית הטבעי של האוכלוסייה, החוקרים יכולים לקבל הבנה מלאה יותר של דינמי שלה...

ישנו גוף הולך וגדל של ספרות חוקר את הקשר בין התנהגויות שונות בתוך חיות בודדות באוכלוסיית נתונה. עמותות אלה מכונות תסמונות התנהגותיות, והמדידות כוללות בדרך כלל תעוזה, תוקפנות, התנהגות גישוש, ו חברותיות 1-5. תסמונות התנהגותיות הן בעלי ערך עבור שני סיבות ישירות ועקיפות. ישירות, ידע של תסמונות התנהגותיות יכול לספק תמונה מלאה יותר של תורת האבולוציה, מבנה האוכלוסייה, ואת הדינמיקה באוכלוסייה 3. בעקיפין, ידע לגבי עמותות התנהגותי עשוי להודיע ​​שדות לכמת התנהגות כגון פרמקולוגיה 6, טוקסיקולוגיה 7, גנטיקה התנהגותית 8,9, ו אנדוקרינולוגיה 10. בגלל יתרונות ישירים ועקיפים אלה, ידע מוגבר תסמונות התנהגותיות הוא בעל ערך במיוחד אורגניזמים מודל נפוצים כמו דג הזברה. מחקרים שעשו שימוש דג הזברה נמצאים iדיסציפלינות נה, כולל ניתוח של תסמונות התנהגותיות 11-13. כדי לקדם את הידע במחקר תסמונת התנהגותית, ובגלל דיסציפלינות אחרות גם להשתמש במדידות התנהגותי בדיקת שערות, תיאורים אמינים ותמציתיים של התנהגות נדרשים עבור ניתוחים ופרשנויות תקפים ופרוטוקולים סטנדרטיים יקלו השוואות בין-מחקר בתוך מינים. פרוטוקול שלנו פותחה כדי למדוד תסמונת התנהגותית תעוזה-תוקפנות-shoaling בקרב אוכלוסיה של דג הזברה מעבדה שגדלו 14. עם זאת, הבסיס של הפרוטוקול (מעקב דגים בודדים, הבטחה אקראית ראוי, ניתוחים מתאימים) ניתן לשנות בקלות עבור מגוון רחב של אמצעי התנהגותיים אלטרנטיביים. בנוסף, התעוזה, תוקפנות, או מבחני shoaling ניתן להפעיל בנפרד עבור הבדיקות של השערות ברורות. לכן, בזמן שהוא המטרה שלנו היא לתאר כיצד לערוך מחקר תסמונת התנהגותית ופרוטוקול עבור l הפרט מוצלחמדידת איבל התנהגותי, כל היבט של הליך זה יכול לעמוד לבד. הספרות על תסמונות התנהגותיות חובקת מספר קבוצות טקסונומיות, מן פרוקי רגליים לבני אדם 4 ו, על מנת למדוד תסמונת התנהגותית, לפחות בשני קשרים ההתנהגותיים חייבים לכמת. למרבה הצער, יש לעתים קרובות עקבי מעט המבחנים המשמשים לכמת את המדידה התנהגותי פני הצירים של התנהגות. לדוגמא, בדגים, תעוזה ניתן למדוד באמצעות מבחני מבוך T, מבחני שדה פתוח, או הקדמה של רומן או גירוי חוץ 15. מחקרי תוקפנות בדגים עלולים להיות כרוכים אינטראקציות הדיאדה, מבחני גירוי וידאו, או מבחנים מחימר 12,16,17. כמו כן, ניתוח של התנהגות shoaling, אשר בדרך כלל כרוכות מדידת העדפת shoalmate, ניתן לבצע סוגים שונים של טנקים, עם שיטות שונות כדי לקבוע עמותת זמן 21-23. בפרוטוקול זה תת-סדרה ספציפית של overaרפרטואר assay ll התנהגותי מוצג. באופן ספציפי, פרוטוקול זה מציג מתודולוגיה כדי לעקוב אחר אנשים דרך תעוזה, תוקפנות, מבחני shoaling באופן המאפשר השוואות בתוך אנשים כדי לקבוע אם המושוות עקביות בכל יחידים בתוך אוכלוסייה. ביצענו פרוטוקול זה עם דג הזברה ציקלידים להרשיע (Amatitlania nigrofasciata) במחקרים קודמים 14,18, וזה יעבוד עם כל דג מים מתוקים בגודל דומה. מבחני תעוזה נערכים בתוך טנק טרפז 1.5 ליטר כי יש קו אופקי המגדירה אזורים בגודל שווה בחלקים העליונים ותחתונים של הטנק. התנהגויות לכמת כוללות את מספר מעברים ידי הדג הבדיקה בין החלק העליון והתחתון של הטנק, הזמן המושקע בכל חלק, מספר חצים, ואת חביון להיכנס החלק העליון. את assay התוקפנות מתבצע בתוך טנק מלבן 19L הכוללמראה אינץ 3 אינץ &#39;x 5 נוטה בסביבות 22 מעלות ממוקמות בפינה השמאלית התחתונה של הטנק 19. התנהגויות לכמת לכלול את הסכום הכולל של זמן מושקע על ידי דג יעד אינטראקציה עם המראה 20, יחד עם אינדיקטורים אגרסיווי ספציפית - מספר העקיצות, מציג לרוחב, חצים בין דגי המבחן וההשתקפות שלו. לקבלת מידע אודות מחוונים הספציפיים אלה, עקיצות מוגדרות מזנקים מהיר לעבר המראה עם פה פעור, מציג לרוחב מוגדר הלוהט של לרוחב, חזה, אנאליים הגבה סנפירים לכיוון המראה, וחיצים הם כל תנועות בלתי יציבות, כי הם לא מכוון במראה. לבסוף, assay shoaling מכמת את ההתנהגות של דג בדיקה בתא במרכז טנק תאי-תלת. תאי דופן האקווריום הם או ריקים, או להכיל &quot;נחיל יעד&quot; של דגים, ובפעם דגי המבחן מבלים ליד כל תא בצד נמדד 21-23. ציון כללי אחד, המכונה וויט&#39;לGTH של Shoaling (SOS), מחושב לגבי כל דג בדיקה אישית, ספציפית לגירויים, והוא יכול לשמש במורד זרם מנתח 14. כל ההתנהגויות הם הבקיעו ידי צופה יחיד, או צופה מרובה באמצעות תוכנת כימות התנהגותי חינם המכונית 24 JWatcher. בדיקת הנוכחות של תסמונת התנהגותית היא מאמץ סטטיסטי בעיקר, ומומלץ לעקוב אחר ההנחיות כפי שהוצג על ידי Budeav 2010 25. באופן ספציפי, מומלץ לבצע ניתוח מרכיבים עיקרי (PCA) על נתונים מרוכזים normed שבו תשומות הן הווקטורים של ההתנהגויות של פרט מבחנים עם מדידות התנהגותי מרובות (כלומר, תעוזה ותוקפנות). את PCA, שבוצעה על מטריצת מתאם, מפחיתה את ממדיות של המדידות התנהגותיות, ובכך מחלצת את הידע החשוב ביותר המסביר ברוב של הווריאציה. הרכיבים שחולצו לאחר מכן ניתן לפרש basאד על מקדמי מתאם גבוה עבור התנהגותם האישית של עשרות ריבית רגרסיה ניתן לחלץ עבור כל אדם על בסיס של רכיבים ההסבר. עשרות רגרסיה אלה לאחר מכן ניתן בהשוואה למדידה SOS ומדידות שאינם התנהגותיות שונות אחרות כגון גודל או מין דגים. עבודה זו יושמה במחקר של תסמונות התנהגותיות דג זברה שבה תסמונת ההתנהגותיות מין שקיימת בין תעוזת shoaling 14 התגלתה. במצב זה, גברי דג זברה נועזים נוטים יותר לקשר עם מינים אגרסיביים יותר, גדולים יותר (ד albolineatus), אך קשר זה הולך לאיבוד אצל נקבות. עבודה זו יושמה גם במחקר של קרובי משפחה לנוער להרשיע ציקליד (Amatitlania nigrofasciata) 18 שבה תסמונת התנהגותית לא התגלה, פוטנציאל המציין פלסטיות התנהגותית של מינים. לכן, בפרוטוקול הבא מוצג wה- i מטרה של המגדירה את האופי שלושה מבחנים ספציפיים (תעוזה, תוקפנות, shoaling) במסגרת לימוד תסמונת התנהגותית ברמת פרט.

<table border="0" cellpadding="0" cellspacing="0" width="783">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Zebrafish Rack System</td>
			<td style="width:224px;">Aquaneering Inc</td>
			<td style="width:119px;">Cat. # ZS550</td>
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			<td height="21" style="height:21px;">Pet Valu Tropical Fish Food, 224.0g</td>
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			<td height="21" style="height:21px;">1.5 L Trapezoidal Tank</td>
			<td>Pentair Aquatic Ecosystems</td>
			<td>Cat. # itsts-a</td>
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			<td height="21" style="height:21px;">19L rectangular tank</td>
			<td>That Fish Place</td>
			<td align="right">211932</td>
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			<td height="21" style="height:21px;">76L rectangular tank</td>
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			<td align="right">212180</td>
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			<td height="21" style="height:21px;">Hitachi KP-D20A CCD Camera</td>
			<td>Prescott&#39;s, Inc.</td>
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		<tr height="21">
			<td height="21" style="height:21px;">Nikon AF Nikkor 35-105mm f/305~4.5s MACRO lens</td>
			<td>Nikon Corporation</td>
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			<td height="21" style="height:21px;">Jwatcher</td>
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			<td height="21" style="height:21px;">SPSS Statistics Base</td>
			<td>IBM</td>
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			<td height="21" style="height:21px;">R</td>
			<td>The R Foundation</td>
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			<td></td>
		</tr>
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</table>

boldness, aggression, and shoaling assays for zebrafish behavioral syndromes

תסמונת התנהגותית מתקיימת כאשר התנהגויות מסוימות אינטראקציה תחת בהקשרים שונים. דג הזברה הייתה נבדקת במחקרים אחרונים וזה חשוב לתקנן פרוטוקולים כדי להבטיח ניתוחים ופרשנויות נכונים. במחקרים הקודמים שלנו, אנו מודדים תעוזה ידי ניטור סדרה של התנהגויות (זמן קרוב לפני שטח, חביון מעברים, מספר מעברים, חצים) בתוך טנק טרפז 1.5 ליטר. כמו כן, אנו לכמת תוקפנות על ידי עקיצות התבוננות, מציג לרוחב, חצים, וזמן ליד מראה נוטה בתוך טנק 19 L מלבני. על ידי חלוקת טנק 76 L לשלישים, גם בדקנו העדפות shoaling. את assay shoaling הוא assay להתאמה אישית והוא יכול להיות מותאם עבור השערות ספציפיות. עם זאת, פרוטוקולים עבור assay זה גם חייבים להיות אחידים, אך גמיש מספיק עבור התאמה אישית. במחקרים קודמים, תא הסוף היו או ריק, כלול 5 או 10 דג זברה, או 5 danios פנינה (ד albolineatus). ב folגעיית כתב יד, אנו מציגים פרוטוקול מפורט ונתוני נציגים שמלווים יישומים מוצלחים של הפרוטוקול, אשר יאפשר לשכפול ניסויי תסמונת התנהגותית.

בהתאם לאופי של המחקר, ופרוטוקול מסוים המועסק, תוצאות ברורות כמה אפשריות בניסוי תסמונות התנהגותיות. הטבלאות הבאות ודמויות, בהם מצוין, מעובדים מנתונים במחקר הקודם שלנו שפורסם בכתב העת ההתנהגות תהליכים 14 וכתב העת דג הזברה 17. כשההצעה (כמתוא?...

Watch this Scientific Journal Video about Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes at JoVE.com

Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

תעוזה, תוקפנות, ואת מבחני Shoaling עבור תסמונות התנהגותיות דג הזברה

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is ...

boldness-aggression-shoaling-assays-for-zebrafish-behavioral

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JoVE Journal

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10.2K Views.  Saint Joseph’s University. This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

Video: Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

Intra-Operative Behavioral Tasks in Awake Humans Undergoing Deep Brain Stimulation Surgery

Deep brain stimulation (DBS) is a surgical procedure that directs chronic, high frequency electrical stimulation to specific targets in the brain through implanted electrodes. Deep brain stimulation was first implemented as a therapeutic modality by Benabid et al. in the late 1980s, when he used this technique to stimulate the ventral intermediate nucleus of the thalamus for the treatment of tremor 1. Currently, the procedure is used to treat patients who fail to respond adequately to medical management for diseases such as Parkinson's, dystonia, and essential tremor. The efficacy of this procedure for the treatment of Parkinson's disease has been demonstrated in well-powered, randomized controlled trials 2. Presently, the U.S. Food and Drug Administration has approved DBS as a treatment for patients with medically refractory essential tremor, Parkinson's disease, and dystonia. Additionally, DBS is currently being evaluated for the treatment of other psychiatric and neurological disorders, such as obsessive compulsive disorder, major depressive disorder, and epilepsy. 
DBS has not only been shown to help people by improving their quality of life, it also provides researchers with the unique opportunity to study and understand the human brain. Microelectrode recordings are routinely performed during DBS surgery in order to enhance the precision of anatomical targeting. Firing patterns of individual neurons can therefore be recorded while the subject performs a behavioral task. Early studies using these data focused on descriptive aspects, including firing and burst rates, and frequency modulation 3. More recent studies have focused on cognitive aspects of behavior in relation to neuronal activity 4,5. This article will provide a description of the intra-operative methods used to perform behavioral tasks and record neuronal data with awake patients during DBS cases. Our exposition of the process of acquiring electrophysiological data will illuminate the current scope and limitations of intra-operative human experiments.

Deep brain stimulation surgery offers a unique opportunity to examine information encoding in the awake human brain. This article will describe intra-operative methods used to perform cognitive and behavioral tasks while simultaneously acquiring physiological data such as EMG, single-unit neuronal activity and/or local field potentials.

תוך Operative משימות ההתנהגות בבני אדם ער שעברו ניתוח עמוק גירוי המוח

Deep brain stimulation (DBS) is a surgical procedure that directs chronic, high frequency electrical stimulation to specific targets in the brain through ...

Using Visual and Narrative Methods to Achieve Fair Process in Clinical Care

The Institute of Medicine has targeted patient-centeredness as an important area of quality improvement. A major dimension of patient-centeredness is respect for patient's values, preferences, and expressed needs. Yet specific approaches to gaining this understanding and translating it to quality care in the clinical setting are lacking. From a patient perspective quality is not a simple concept but is best understood in terms of five dimensions: technical outcomes; decision-making efficiency; amenities and convenience; information and emotional support; and overall patient satisfaction. Failure to consider quality from this five-pronged perspective results in a focus on medical outcomes, without considering the processes central to quality from the patient's perspective and vital to achieving good outcomes. In this paper, we argue for applying the concept of fair process in clinical settings. Fair process involves using a collaborative approach to exploring diagnostic issues and treatments with patients, explaining the rationale for decisions, setting expectations about roles and responsibilities, and implementing a core plan and ongoing evaluation. Fair process opens the door to bringing patient expertise into the clinical setting and the work of developing health care goals and strategies. This paper provides a step by step illustration of an innovative visual approach, called photovoice or photo-elicitation, to achieve fair process in clinical work with acquired brain injury survivors and others living with chronic health conditions. Applying this visual tool and methodology in the clinical setting will enhance patient-provider communication; engage patients as partners in identifying challenges, strengths, goals, and strategies; and support evaluation of progress over time. Asking patients to bring visuals of their lives into the clinical interaction can help to illuminate gaps in clinical knowledge, forge better therapeutic relationships with patients living with chronic conditions such as brain injury, and identify patient-centered goals and possibilities for healing. The process illustrated here can be used by clinicians, (primary care physicians, rehabilitation therapists, neurologists, neuropsychologists, psychologists, and others) working with people living with chronic conditions such as acquired brain injury, mental illness, physical disabilities, HIV/AIDS, substance abuse, or post-traumatic stress, and by leaders of support groups for the types of patients described above and their family members or caregivers.

This paper illustrates an innovative visual approach (photovoice or photo-elicitation) to achieve fair process in clinical care for patients living with chronic health conditions, illuminate gaps in clinical knowledge, forge better therapeutic relationships, and identify patient-centered goals and possibilities for healing.

באמצעות שיטות חזותית הנרטיב להשגת הליך הוגן בטיפול קליני

The Institute of Medicine has targeted patient-centeredness as an important area of quality improvement. A major dimension of patient-centeredness is ...

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

סמנים ביולוגיים במודל חיה לחשיפה קושרת עצבית, המטולוגית, והתנהגות של PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

Eye Tracking Young Children with Autism

The rise of accessible commercial eye-tracking systems has fueled a rapid increase in their use in psychological and psychiatric research. By providing a direct, detailed and objective measure of gaze behavior, eye-tracking has become a valuable tool for examining abnormal perceptual strategies in clinical populations and has been used to identify disorder-specific characteristics1, promote early identification2, and inform treatment3. In particular, investigators of autism spectrum disorders (ASD) have benefited from integrating eye-tracking into their research paradigms4-7. Eye-tracking has largely been used in these studies to reveal mechanisms underlying impaired task performance8 and abnormal brain functioning9, particularly during the processing of social information1,10-11. While older children and adults with ASD comprise the preponderance of research in this area, eye-tracking may be especially useful for studying young children with the disorder as it offers a non-invasive tool for assessing and quantifying early-emerging developmental abnormalities2,12-13. Implementing eye-tracking with young children with ASD, however, is associated with a number of unique challenges, including issues with compliant behavior resulting from specific task demands and disorder-related psychosocial considerations. In this protocol, we detail methodological considerations for optimizing research design, data acquisition and psychometric analysis while eye-tracking young children with ASD. The provided recommendations are also designed to be more broadly applicable for eye-tracking children with other developmental disabilities. By offering guidelines for best practices in these areas based upon lessons derived from our own work, we hope to help other investigators make sound research design and analysis choices while avoiding common pitfalls that can compromise data acquisition while eye-tracking young children with ASD or other developmental difficulties.

Eye tracking has long been used to study gaze patterns in typically-developing individuals, but recent technological advancements have made its use with clinical populations, including autism, more feasible. While eye-tracking young children with autism can offer insight into early symptom manifestations, it involves methodological challenges. Suggestions for best practices are provided.

מעקב עיניים ילדים צעירים עם אוטיזם

The rise of accessible commercial eye-tracking systems has fueled a rapid increase in their use in psychological and psychiatric research. By providing a ...

Processing of Primary Brain Tumor Tissue for Stem Cell Assays and Flow Sorting

Brain tumors are typically comprised of morphologically diverse cells that express a variety of neural lineage markers. Only a relatively small fraction of cells in the tumor with stem cell properties, termed brain tumor initiating cells (BTICs), possess an ability to differentiate along multiple lineages, self-renew, and initiate tumors in vivo. We applied culture conditions originally used for normal neural stem cells (NSCs) to a variety of human brain tumors and found that this culture method specifically selects for stem-like populations. Serum-free medium (NSC) allows for the maintenance of an undifferentiated stem cell state, and the addition of bFGF and EGF allows for the proliferation of multi-potent, self-renewing, and expandable tumorspheres.
To further characterize each tumor's BTIC population, we evaluate cell surface markers by flow cytometry. We may also sort populations of interest for more specific characterization. Self-renewal assays are performed on single BTICs sorted into 96 well plates; the formation of tumorspheres following incubation at 37 &deg;C indicates the presence of a stem or progenitor cell. Multiple cell numbers of a particular population can also be sorted in different wells for limiting dilution analysis, to analyze self-renewal capacity. We can also study differential gene expression within a particular cell population by using single cell RT-PCR.
The following protocols describe our procedures for the dissociation and culturing of primary human samples to enrich for BTIC populations, as well as the dissociation of tumorspheres. Also included are protocols for staining for flow cytometry analysis or sorting, self-renewal assays, and single cell RT-PCR.

The identification of brain tumor initiating cells (BTICs), the rare cells within a heterogeneous tumor possessing stem cell properties, provides new insights into human brain tumor pathogenesis. We have refined specific culture conditions to enrich for BTICs, and we routinely use flow cytometry to further enrich these populations. Self-renewal assays and transcript analysis by single cell RT-PCR can subsequently be performed on these isolated cells.

עיבוד של רקמת גידול במוח העיקרית למבחנים בתאי גזע וזרימת מיון

Brain tumors are typically comprised of morphologically diverse cells that express a variety of neural lineage markers. Only a relatively small fraction ...

Screening for Melanoma Modifiers using a Zebrafish Autochthonous Tumor Model

Genomic studies of human cancers have yielded a wealth of information about genes that are altered in tumors1,2,3. A challenge arising from these studies is that many genes are altered, and it can be difficult to distinguish genetic alterations that drove tumorigenesis from that those arose incidentally during transformation. To draw this distinction it is beneficial to have an assay that can quantitatively measure the effect of an altered gene on tumor initiation and other processes that enable tumors to persist and disseminate. Here we present a rapid means to screen large numbers of candidate melanoma modifiers in zebrafish using an autochthonous tumor model4 that encompasses steps required for melanoma initiation and maintenance. A key reagent in this assay is the miniCoopR vector, which couples a wild-type copy of the mitfa melanocyte specification factor to a Gateway recombination cassette into which candidate melanoma genes can be recombined5. The miniCoopR vector has a mitfa rescuing minigene which contains the promoter, open reading frame and 3'-untranslated region of the wild-type mitfa gene. It allows us to make constructs using full-length open reading frames of candidate melanoma modifiers. These individual clones can then be injected into single cell Tg(mitfa:BRAFV600E);p53(lf);mitfa(lf)zebrafish embryos. The miniCoopR vector gets integrated by Tol2-mediated transgenesis6 and rescues melanocytes. Because they are physically coupled to the mitfa rescuing minigene, candidate genes are expressed in rescued melanocytes, some of which will transform and develop into tumors. The effect of a candidate gene on melanoma initiation and melanoma cell properties can be measured using melanoma-free survival curves, invasion assays, antibody staining and transplantation assays.

A rapid way to screen for melanoma modifiers using a zebrafish autochthonous tumor model is presented. It takes advantage of the miniCoopR vector which allows for expression of candidate melanoma genes in melanocytes. A method to obtain melanoma-free survival curves, an invasion assay, a protocol for antibody staining of scale melanocytes and a melanoma transplantation assay are described.

הקרנה למכפילים מלנומה תוך שימוש במודל גידול כבני מקום דג זברה

Genomic studies of  human cancers have yielded a wealth of information about genes that are altered  in tumors1,2,3. A challenge arising from these ...

Adaptation of Semiautomated Circulating Tumor Cell (CTC) Assays for Clinical and Preclinical Research Applications

The majority of cancer-related deaths occur subsequent to the development of metastatic disease. This highly lethal disease stage is associated with the presence of circulating tumor cells (CTCs). These rare cells have been demonstrated to be of clinical significance in metastatic breast, prostate, and colorectal cancers. The current gold standard in clinical CTC detection and enumeration is the FDA-cleared CellSearch system (CSS). This manuscript outlines the standard protocol utilized by this platform as well as two&#160;additional adapted protocols that describe the detailed process of user-defined marker optimization for protein characterization of patient CTCs and a comparable protocol for CTC capture in very low volumes of blood, using standard CSS reagents, for studying in vivo preclinical mouse models of metastasis. In addition, differences in CTC quality between healthy donor blood spiked with cells from tissue culture versus patient blood samples are highlighted. Finally, several commonly discrepant items that can lead to CTC misclassification errors are outlined. Taken together, these protocols will provide a useful resource for users of this platform interested in preclinical and clinical research pertaining to metastasis and CTCs.

Adaptation of Semiautomated Circulating Tumor Cell CTC Assays for Clinical and Preclinical Research Applications

Circulating tumor cells (CTCs) are prognostic in several metastatic cancers. This manuscript describes the gold standard CellSearch system (CSS) CTC enumeration platform and highlights common misclassification errors. In addition, two&#160;adapted protocols are described for user-defined marker characterization of CTCs and CTC enumeration in preclinical mouse models of metastasis using this technology.

עיבוד של semiautomated מחזור תא גידול מבחני (CTC) ליישומים קליניים ומחקר הקליני

The majority of cancer-related deaths occur subsequent to the development of metastatic disease. This highly lethal disease stage is associated with the ...

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone resorption. Osteoclasts can be generated in vitro from monocyte/macrophage precursor cells in the presence of certain cytokines, which promote survival and differentiation. Here, both in vivo and in vitro techniques are demonstrated, which allow scientists to study different cytokine contributions towards osteoclast differentiation, signaling, and activation. The minicircle DNA delivery gene transfer system provides an alternative method to establish an osteoporosis-related model is particularly useful to study the efficacy of various pharmacological inhibitors in vivo. Similarly, in vitro culturing protocols for producing osteoclasts from human precursor cells in the presence of specific cytokines enables scientists to study osteoclastogenesis in human cells for translational applications. Combined, these techniques have the potential to accelerate drug discovery efforts for osteoclast-specific targeted therapeutics, which may benefit millions of osteoporosis and arthritis patients worldwide.

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation of precursor cells into osteoclasts is regulated by cytokines and growth factors. Here, a novel gene transfer technique for differentiation of osteoclasts in vivo and cell culture protocols for differentiating precursor cells into osteoclasts in vitro as a method to study the effects of cytokines on osteoclastogenesis are described.

רומן<em&gt; In vivo</em&gt; טכניקה וין העברה<em&gt; במבחנה</emמבחני תא בהתבסס&gt; לחקר אובדן עצם בפרעות השלד והשרירים

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone ...

A Novel Model of Mild Traumatic Brain Injury for Juvenile Rats

Despite growing evidence that childhood represents a major risk period for mild traumatic brain injury (mTBI) from sports-related concussions, motor vehicle accidents, and falls, a reliable animal model of mTBI had previously not been developed for this important aspect of development. The modified weight-drop technique employs a glancing impact to the head of a freely moving rodent transmitting acceleration, deceleration, and rotational forces upon the brain. When applied to juvenile rats, this modified weight-drop technique induced clinically relevant behavioural outcomes that were representative of post-concussion symptomology. The technique is a rapidly applied procedure with an extremely low mortality rate, rendering it ideal for high-throughput studies of therapeutics. In addition, because the procedure involves a mild injury to a closed head, it can easily be used for studies of repetitive brain injury. Owing to the simplistic nature of this technique, and the clinically relevant biomechanics of the injury pathophysiology, the modified weight-drop technique provides researchers with a reliable model of mTBI that can be used in a wide variety of behavioural, molecular, and genetic studies.

The modified weight-drop technique is an easy, cost-effective procedure used for the induction of mild traumatic brain injury in juvenile rats. This novel technique produces clinically relevant symptomology that will advance the study of mild traumatic brain injury (mTBI) and concussion.

דגם רומן של טראומטית קל פגיעה מוחית לחולדות לנוער

Despite growing evidence that childhood represents a major risk period for mild traumatic brain injury (mTBI) from sports-related concussions, motor ...

Real-time Cytotoxicity Assays in Human Whole Blood

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with high-throughput cell positioning technology. The targeted tumor cell populations are first preprogrammed to immobilization into an array format, and labeled with green fluorescent cytosolic dyes. Following the cell array formation, antibody drugs are added in combination with human whole blood. Propidium iodide (PI) is then added to assess cell death. The cell array is analyzed with an automatic imaging system. While cytosolic dye labels the targeted tumor cell populations, PI labels the dead tumor cell populations. Thus, the percentage of target cancer cell killing can be quantified by calculating the number of surviving targeted cells to the number of dead targeted cells. With this method, researchers are able to access time-dependent and dose-dependent cell cytotoxicity information. Remarkably, no hazardous radiochemicals are used. The WCA presented here has been tested with lymphoma, leukemia, and solid tumor cell lines. Therefore, WCA allows researchers to assess drug efficacy in a highly relevant ex&#160;vivo condition.

The whole blood cytotoxicity assay (WCA) is a cytotoxicity assay developed by incorporating high-throughput cell positioning technology with fluorescence microscopy and automated image processing. Here, we describe how lymphoma cells treated with an anti-CD20 antibody can be analyzed real-time in human whole blood to provide quantitative cellular cytotoxicity analysis.

בזמן אמת Cytotoxicity מבחני בדם כל אדם

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with ...