Name: prevention of heat stress adverse effects in rats by bacillus subtilis strain
Uploaded: 2016-07-11T14:00:00.000+00:00
Duration: 7 min 57 s
Description: Watch this Scientific Journal Video about Prevention of Heat Stress Adverse Effects in Rats by Bacillus subtilis Strain at JoVE.com

This work was supported by the Auburn University fund FOP 101002-139294-2050.

Alle experimentellen Verfahren wurden von IACUC der Auburn University, Auburn, Alabama genehmigt. 1. Herstellung von Kulturmedien, Geschirr und Bakterienkultur <ol><li> Impfen 10 ml Nährlösung in einem Kolben mit einer einzelnen Kolonie von Bacillus subtilis BSB3 Kultur, über Nacht auf einem Nährstoff - Agar - Platte gewachsen. </li><li> Machen 500 ml Sporulationsmedium 19 (pro Liter: Bacto Nutrient Broth, 8 g; 10% (w / v) KCl, 10 ml; 1,2% (w / v) MgSO4 x 7 H 2 O, 10 ml, Agar, 15 g) und Autoklaven bei 121 ° C für 20 min. Lassen Sie auf 50 o C abkühlen und fügen Sie die folgenden sterilen Lösungen: 1 M Ca (NO 3) 2, 1 ml; 0,01 M MnCl 2, 1 ml; 1 mM FeSO 4, 1 ml. </li><li> Kühlen hergestellte Medium bis 40 o C für 20 min bei Raumtemperatur und gießt 25 ml in jede von 20 sterilen Petrischalen im Sterilschrank. Lassen Sie das Medium über Nacht erstarren. </li><li> Verteilt 0,5 ml Übernachtkultur von Bacillus subtilis BSB3 auf die Oberfläche des Mediums in den Petrischalen. Inkubiere Kultur bei 37 ° C für 5 Tage , bis 90% der Sporulation. Reveal Phase helle Sporen durch die hochauflösende Mikroskopie. </li><li> Ernte Bakterien durch Zugabe von 1 ml steriler phosphatgepufferter Salzlösung (PBS) auf die Platte und Abkratzen Bakterien von einer Oberfläche eines sterilen Zell Spreizer verwendet wird. Bewahren Sie die Suspension in den Kühlschrank stellen, bis sie benötigt. </li><li> Bestätigen Lebensfähigkeit der Bakterien durch 0,1 ml der geeigneten Verdünnung Plattierung (10 -5 bis 10 -6) einer Bakteriensuspension auf Sporulationsmedium Platten gefolgt von einer Inkubation für 18-24 Stunden bei 37 o C. </li><li> Graf Bakterienkolonien auf den Platten eine Kolonie Zähler mit und berechnen den Titer von Bakterien in der getesteten Suspension. Vorbereitung Bakteriensuspension mit der Endkonzentration 1 x 10 8 CFU / ml in PBS. </li></ol> 2. Tiere <ol><li> Verwenden Sie 32 männliche Sprague-Dawley-Ratten (250 - 300 g,). Pflegen Tiere unter spezifischen pathogenfreien Bedingungen mit freiem Zugang zu Standard-Pelletfutter und Wasser. Stellen Sie eine mittlere Temperatur (21 ± 1 ° C), Feuchtigkeit (50 ± 5%) und eine 12 - Stunden Licht-Dunkel - Zyklus. </li></ol> 3. Experimentelles Design <ol><li> Starten Sie Behandlung von Tieren durch orale Gabe 11, nach 2 Tagen der Akklimatisierung. Verwenden Sie Spritze mit der Schlundsonde Nadel. Behandeln Sie 16 Ratten , die mit B. subtilis BSB3 Suspension (1 ml pro Ratte) zweimal täglich für zwei Tage (B. subtilis - Gruppe). Behandeln Sie 16 Ratten , die mit PBS (1 ml pro Ratte) zweimal täglich für zwei Tage (PBS - Gruppe) (Abbildung 1). </li><li> Nach der Behandlung unterteilen jede Gruppe (8 Ratten pro Gruppe): Gruppe 1- Kontrolle (PBS / kein Stress), Gruppe 2 - B subtilis (B. subtilis / kein Stress), Gruppe 3 - Stress (PBS / Hitzestress) und Gruppe 4- B. subtilis + Spannung (B. subtilis / Hitzestress). </li><li> Messen Sie die Temperatur rektal jeder Ratte unter Verwendung eines elektronic digitales Thermometer. Halten Ratten der Gruppen 1 und 2 bei Raumtemperatur für 25 min. Ort Tiere der Gruppen 3 und 4 in einer Klimakammer bei 45 o C und die relative Luftfeuchtigkeit 55% für 25 min. </li><li> Anschließend messen die rektale Temperatur jeder Ratte in allen Gruppen eine elektronische Digitalthermometer verwenden. Legen Sie alle Tiere bei Raumtemperatur für 4 Stunden. </li><li> Anesthetize Ratten mit Isofluran (2-4%) in ein verschließbares Gefäß Anästhesie und beobachten, bis die Ratten tief anästhesiert (keine Antwort auf Zehe Prise). Euthanize Ratten durch schnelle Enthauptung mit einer Guillotine. </li><li> Sammeln Stamm Blut von jeder Ratte 20 (15 - 16 ml) mit den apyrogene Pipetten in apyrogene Röhren Serum zu erhalten und sofort nehmen Proben von Blut (7 ul von jeder Ratte) mit einer Pipette für die mikroskopische Untersuchung. </li><li> Setzen Sie Röhrchen mit dem Blut in einem Kühlschrank für mindestens 30 min Gerinnen zu ermöglichen. Zentrifugenröhrchen bei 20 ° C, 7000 × g für 10 min und schnellentfernen Serum mit einer Pipette. Aliquot Serum von jeder Ratte in 50 ul - Volumina und bei -20 ° C bis zum Test erhalten. </li></ol> 4. Chirurgische Verfahren <ol><li> Cut / trimmen alle Fell aus dem Bauch und Brustkorb der Unterseite der Karkasse mit elektrischen Scheren. Legen Sie die Karkasse in einem sterilen Schrank und behandeln die rasierte Fläche der Karkasse mit 70% Alkohol. </li><li> Verwenden Sie eine sterile Skalpell einen Mittelschnitt des Bauches zu schaffen. Entfernen Leber, mesenterischen Lymphknoten, Milz und Dünndarm von jeder Ratte unter Verwendung von sterilen Pinzetten. </li></ol> 5. Analyse der bakteriellen Translokation <ol><li> Platzieren Sie jede Probe von Leber, mesenterischen Lymphknoten und Milz in vorher gewogene sterilen Röhrchen und wiegen. Berechnen des Gewichts der Probe als eine Differenz zwischen dem Gewicht eines Röhrchens mit einer Probe und das Gewicht eines leeren Rohrs. </li><li> Hinzufügen sterile PBS auf das Rohr eine 1:10 Verdünnung zu erhalten (w / v) der Probe und homogerieren jede Probe mit einem sterilen Homogenisator Glasgewebe eine homogene Suspension 21 zu erhalten. </li><li> Stellen Reihenverdünnungen (10 -1 bis 10 -4) jeder Probe in sterilem PBS. Platte 0,1 ml aller Verdünnungen jedes Gewebes auf die Oberfläche von MacConkey des und 5% Blutagar und Brucella Blutagar mit Hämin (0,005 g / l) und Vitamin K1 (0,01 g / L) Platten. </li><li> Inkubieren MacConkey des und 5% Blutagarplatten aerob und Brucella Blutagarplatten unter anaeroben Bedingungen bei 37 o C 22. </li><li> Graf Kolonien von aeroben Bakterien nach 24 Stunden, und Kolonien von anaeroben Bakterien nach 48 h Inkubation eine Kolonie Zähler verwendet wird. Die Ergebnisse sind als die Anzahl der koloniebildenden Einheiten (CFU) in einem Gramm Gewebe. </li></ol> 6. Die histologische Analyse <ol><li> Entfernen Dünndarm Proben aus jeder Ratte durch die chirurgische Prozedur (Schritte von 4,1 bis 4,2). Fix Proben in Bouin-Fixiermittel, einbetten in Paraffin,Vorbereitung 6 um Abschnitte und Fleckenabschnitte mit Hämatoxylin und Eosin nach Standardverfahren 23. </li><li> Verwenden , um eine hochauflösende Mikroskopsystem für die Höhe der Darmzotten Messung und Gesamtschleimhautdicke in jeder Probe (Vergrßerung 100X) 24. Analysieren 4 Proben von jeder Ratte und machen mindestens zwanzig Messungen in jeder Probe. Express-Ergebnisse in Mikrometer. </li></ol> 7. Zytokin-Assay <ol><li> Verwenden Sie kommerzielle Ratte ELISA-Kits für IL-1β; IL-6; TNF-α; INF-γ und IL-10-Niveaus von Zytokinen in Serum zu messen, gemäß dem Protokoll des Herstellers. </li><li> Generieren von Standardkurven für jeden Probensatz unter Verwendung von Standards für den entsprechenden Kit getestet. Definieren Sie die Ebene von Zytokinen in jeder Probe durch Vergleich der experimentellen Daten mit der entsprechenden Standardkurve. </li></ol> 8. Lipopolysaccharide Assay <ol><li> Analysieren Niveau von LPS in Serum eine Ratte LPS ELISA-Kit nachProtokoll des Herstellers. Schätzung Konzentration von LPS in den Proben durch Vergleich der erhaltenen Werte mit den Werten der Standardkurve. </li></ol> 9. Hohe Auflösung Lichtmikroskopie des Blutes <ol><li> Verwenden Sie das Mikroskopsystem vorher 25,26 beschrieben. Verwenden Sie eine Schwingungsisolationsplattform als Basis für das Mikroskopsystem und Videokamera und Computer 25 , um die Live - Bilder aufzunehmen. Kalibrieren Sie Testbilder einen Richardson Schlitten unter Verwendung von wie zuvor 27 beschrieben. </li><li> Legen 7 ul frisch entnommenem Blut von jeder Ratte auf einem Glasobjektträger, Deckglas, Fotografie und aufzeichnen zehn Bildrahmen von 72 × 53,3 &amp; mgr; m 2 in jeder Probe (Vergrößerung 1,700X). </li><li> Messen Sie die Konzentration von Vesikeln mit einer Software, die Direktsicht optische Bilder in Echtzeit mit hoher Auflösung liefert. Untersuchen mindestens 20 Bildrahmen für jeden experimentellen Zustand 28. </li></ol> 10. StatisTics <ol><li> Express alle Werte als Mittelwert und Standardabweichung. Führen Sie eine statistische Analyse und Grafik Plotten. Analysieren Sie die Ergebnisse mit den beiden Probe t-Test, stellen Sie das Signifikanzniveau bei 0,05 statistische Signifikanz zu definieren. </li></ol>

<ol>
	<li>Crandall, C. G., Gonzalez-Alonso, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiovascular+function+in+the+heat-stressed+human.">Cardiovascular function in the heat-stressed human.</a> Acta Physiol. 199, 407-423 (2010).</li><li>Lambert, G. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+gastrointestinal+permeability+in+exertional+heatstroke.">Role of gastrointestinal permeability in exertional heatstroke.</a> Exerc. Sport Sci. Rev. 32, 185-190 (2004).</li><li>Yu, J., 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=Effect+of+heat+stress+on+the+porcine+small+intestine:+A+morphological+and+gene+expression+study.">Effect of heat stress on the porcine small intestine: A morphological and gene expression study.</a> Comp. Biochem. Physiol. A-Mol. Integr. Physiol. 156, 119-128 (2010).</li><li>Moseley, P. L., Gisolfi, C. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+Frontiers+in+Thermoregulation+and+Exercise.">New Frontiers in Thermoregulation and Exercise.</a> Sports Med. 16, 163-167 (1993).</li><li>Lambert, G. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stress-induced+gastrointestinal+barrier+dysfunction+and+its+inflammatory+effects.">Stress-induced gastrointestinal barrier dysfunction and its inflammatory effects.</a> J. Anim. Sci. 87, 101-108 (2009).</li><li>Berg, A., Muller, H. M., Rathmann, S., Deibert, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+gastrointestinal+system+-+An+essential+target+organ+of+the+athlete's+health+and+physical+performance.">The gastrointestinal system - An essential target organ of the athlete's health and physical performance.</a> Exerc. Immunol. Rev. 5, 78-95 (1999).</li><li>Khan, M. W., 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=Microbes,+intestinal+inflammation+and+probiotics.">Microbes, intestinal inflammation and probiotics.</a> Expert Rev Gastroent. 6, 81-94 (2012).</li><li>Quigley, E. M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prebiotics+and+Probiotics:+Their+Role+in+the+Management+of+Gastrointestinal+Disorders+in+Adults.">Prebiotics and Probiotics: Their Role in the Management of Gastrointestinal Disorders in Adults.</a> Nutr Clin Pract. 27, 195-200 (2012).</li><li>Gore, C., 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=Treatment+and+secondary+prevention+effects+of+the+probiotics+Lactobacillus+paracasei+or+Bifidobacterium+lactis+on+early+infant+eczema:+randomized+controlled+trial+with+follow-up+until+age+3+years.">Treatment and secondary prevention effects of the probiotics Lactobacillus paracasei or Bifidobacterium lactis on early infant eczema: randomized controlled trial with follow-up until age 3 years.</a> Clin Exp Allergy. 42, 112-122 (2012).</li><li>Gomes, A. C., Bueno, A. A., de Souza, R. G. M., Mota, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gut+microbiota,+probiotics+and+diabetes.">Gut microbiota, probiotics and diabetes.</a> Nutr. J. 13, (2014).</li><li>Eutamene, H., 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=Synergy+between+Lactobacillus+paracasei+and+its+bacterial+products+to+counteract+stress-induced+gut+permeability+and+sensitivity+increase+in+rats.">Synergy between Lactobacillus paracasei and its bacterial products to counteract stress-induced gut permeability and sensitivity increase in rats.</a> J. Nutr. 137, 1901-1907 (2007).</li><li>Ait-Belgnaoui, 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=Lactobacillus+farciminis+treatment+attenuates+stress-induced+overexpression+of+Fos+protein+in+spinal+and+supraspinal+sites+after+colorectal+distension+in+rats.">Lactobacillus farciminis treatment attenuates stress-induced overexpression of Fos protein in spinal and supraspinal sites after colorectal distension in rats.</a> Neurogastroenterol. Motil. 21, 585-593 (2009).</li><li>Fujiya, M., 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=The+Bacillus+subtilis+quorum-sensing+molecule+CSF+contributes+to+intestinal+Homeostasis+via+OCTN2,+a+host+cell+membrane+transporter.">The Bacillus subtilis quorum-sensing molecule CSF contributes to intestinal Homeostasis via OCTN2, a host cell membrane transporter.</a> Cell Host Microbe. 1, 299-308 (2007).</li><li>Cutting, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bacillus+probiotics.">Bacillus probiotics.</a> Food Microbiol. 28, 214-220 (2011).</li><li>Pinchuk, I. V., 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=In+vitro+anti-Helicobacter+pylori+activity+of+the+probiotic+strain+Bacillus+subtilis+3+is+due+to+secretion+of+antibiotics.">In vitro anti-Helicobacter pylori activity of the probiotic strain Bacillus subtilis 3 is due to secretion of antibiotics.</a> Antimicrob Agents Ch. 45, 3156-3161 (2001).</li><li>Sorokulova, I. B., Kirik, D. L., Pinchuk, I. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Probiotics+against+Campylobacter+pathogens.">Probiotics against Campylobacter pathogens.</a> J. Travel Med. 4, 167-170 (1997).</li><li>Gracheva, N. M., 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=The+efficacy+of+the+new+bacterial+preparation+biosporin+in+treating+acute+intestinal+infections.">The efficacy of the new bacterial preparation biosporin in treating acute intestinal infections.</a> Zh Mikrobiol Epidemiol Immunobiol. , 75-77 (1996).</li><li>Horosheva, T. V., Vodyanoy, V., Sorokulova, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+of+Bacillus+probiotics+in+prevention+of+antibiotic-associated+diarrhoea:+a+randomized,+double-blind,+placebo-controlled+clinical+trial.">Efficacy of Bacillus probiotics in prevention of antibiotic-associated diarrhoea: a randomized, double-blind, placebo-controlled clinical trial.</a> JMM Case Report. 1, (2014).</li><li>Sorokulova, I. B., Krumnow, A. A., Pathirana, S., Mandell, A. J., Vodyanoy, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+Methods+for+Storage+Stability+and+Release+of+Bacillus+Spores.">Novel Methods for Storage Stability and Release of Bacillus Spores.</a> Biotechnol. Prog. 24, 1147-1153 (2008).</li><li>Vogel, H. G., Vogel, W. H., Vogel, H. G., Vogel, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug+Discovery+and+Evaluation:+Pharmacological+Assays.">Drug Discovery and Evaluation: Pharmacological Assays.</a> eds H.G. Vogel &amp; W.H. Vogel. , 658-659 (1997).</li><li>Bailey, M. T., Engler, H., Sheridan, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stress+induces+the+translocation+of+cutaneous+and+gastrointestinal+microflora+to+secondary+lymphoid+organs+of+C57BL/6+mice.">Stress induces the translocation of cutaneous and gastrointestinal microflora to secondary lymphoid organs of C57BL/6 mice.</a> J. Neuroimmunol. 171, 29-37 (2006).</li><li>Rakoff-Nahoum, S., Paglino, J., Eslami-Varzaneh, F., Edberg, S., Medzhitov, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recognition+of+commensal+microflora+by+toll-like+receptors+is+required+for+intestinal+homeostasis.">Recognition of commensal microflora by toll-like receptors is required for intestinal homeostasis.</a> Cell. 118, 229-241 (2004).</li><li>Leon, L. R., Blaha, M. D., DuBose, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time+course+of+cytokine,+corticosterone,+and+tissue+injury+responses+in+mice+during+heat+strain+recovery.">Time course of cytokine, corticosterone, and tissue injury responses in mice during heat strain recovery.</a> J. Appl. Physiol. 100, 1400-1409 (2006).</li><li>Ribeiro, S. R., 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=Weight+loss+and+morphometric+study+of+intestinal+mucosa+in+rats+after+massive+intestinal+resection.+Influence+of+a+glutamine-enriched+diet.">Weight loss and morphometric study of intestinal mucosa in rats after massive intestinal resection. Influence of a glutamine-enriched diet.</a> Rev. Hosp. Cl&#236;n. Fac. Med. S. Paulo. 59, 349-356 (2004).</li><li>Vainrub, A., Pustovyy, O., Vodyanoy, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resolution+of+90+nm+(lambda/5)+in+an+optical+transmission+microscope+with+an+annular+condenser.">Resolution of 90 nm (lambda/5) in an optical transmission microscope with an annular condenser.</a> Opt Lett. 31, 2855-2857 (2006).</li><li>Moore, T., Globa, L., Pustovyy, O., Vodyanoy, V., Sorokulova, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oral+administration+of+Bacillus+subtilis+strain+BSB3+can+prevent+heat+stress-related+adverse+effects+in+rats.">Oral administration of Bacillus subtilis strain BSB3 can prevent heat stress-related adverse effects in rats.</a> J Appl Microbiol. 117, 1463-1471 (2014).</li><li>Richardson, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Test+slides:+Diatoms+to+divisions-What+are+you+looking+at.">Test slides: Diatoms to divisions-What are you looking at.</a> Proc Roy Microsc Soc. 22, 3-9 (1988).</li><li>Moore, T., Sorokulova, I., Pustovyy, O., Globa, L., Vodyanoy, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microscopic+evaluation+of+vesicles+shed+by+rat+erythrocytes+at+elevated+temperatures.">Microscopic evaluation of vesicles shed by rat erythrocytes at elevated temperatures.</a> J Therm Biol. 38, 487-492 (2013).</li><li>Leon, L. R., Helwig, B. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heat+stroke:+Role+of+the+systemic+inflammatory+response.">Heat stroke: Role of the systemic inflammatory response.</a> J. Appl. Physiol. 109, 1980-1988 (2010).</li><li>Kumar, Y., Chawla, A., Tatu, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heat+Shock+Protein+70+as+a+Biomarker+of+Heat+Stress+in+a+Simulated+Hot+Cockpit.">Heat Shock Protein 70 as a Biomarker of Heat Stress in a Simulated Hot Cockpit.</a> Aviat Space Env Med. 74 (7), 711-716 (2007).</li></ol>

Die Autoren erklären, dass sie keine finanziellen Interessen haben.

Die Exposition gegenüber hohen Temperaturen führt zu schweren gesundheitlichen Bedingungen 29. Prävention und Früherkennung von Komplikationen zu Hitzestress im Zusammenhang von entscheidender Bedeutung sind 30. Das dargestellte Protokoll kann die Wirksamkeit verschiedener Ansätze zur Verhinderung von Hitzestress nachteiligen Wirkungen zu bewerten, verwendet werden. Kritische Schritte in diesem Protokoll sind: die Wärmebehandlungsverfahren (45 <...

Different stressors affect human and animal health. Temperature is one of the most stressful factors, causing chronic, acute and even lethal illnesses1. Changes in intestinal morphology and a loss of gut barrier integrity after heat stress were documented in many cases2,3. This protective barrier is responsible for defense against translocation of gut bacteria and their toxins, in particular lipopolysaccharides (LPS), from the lumen to the internal circulation4,5. Stability of the gut microbiota significantly influences intestinal barrier function, the immune response of the host, and tolerance to stress conditions6. Thus, modulation of the intestinal microbiota can provide a novel approach for prevention of stress-related adverse effects. 
Beneficial bacteria have been used as a promising strategy to modify the gut microbiota for successful management of various clinical conditions, such as diarrhea, inflammatory bowel disease, atopic dermatitis, metabolic disorders7-10. Probiotic effects of beneficial bacteria include production of essential metabolites to support intestinal health, stimulation of the immune system, promotion of lactose tolerance, restoration of epithelial dysfunction. Probiotics were effective in prevention of the stress-related complications in vitro and in animal models11,12. 
Researchers have given more attention to Bacillus bacteria as probiotics because these bacteria support intestinal homeostasis13 and have beneficial effects on the host14. Our previous data demonstrated high efficacy of Bacillus probiotics against pathogens in vitro15,16 and in clinical trials17,18. Here, we aim to evaluate the preventive effect of B. subtilis BSB3 strain against complications after heat stress.

<table><tbody><tr><td>Phosphate buffered saline (PBS)</td><td>Sigma-Aldrich,&amp;nbsp;St. Louis, MO</td><td>P4417</td><td></td></tr><tr><td>Ethyl Alcohol&amp;nbsp;</td><td>Pharmco products Inc. Brookfield, CT, USA&amp;nbsp;</td><td>64-17-5</td><td></td></tr><tr><td>Agar</td><td>VWR</td><td>97064-334</td><td></td></tr><tr><td>MacConkey agar plates</td><td>VWR</td><td>470180-742</td><td></td></tr><tr><td>5% blood agar plates</td><td>VWR</td><td>89405-024</td><td></td></tr><tr><td>Brucella blood agar plate with Hemin, and Vitamin K</td><td>VWR</td><td>89405-032</td><td></td></tr><tr><td>Bouin&amp;rsquo;s Fixative</td><td>Electron Microscopy Sciences, Hatfield, PA, USA</td><td>15990</td><td></td></tr><tr><td>IL-10 Rat ELISA kit</td><td>Invitrogen, Camarillo, CA, USA</td><td>KRC0101</td><td></td></tr><tr><td>IL-1beta Rat ELISA Kit</td><td>Invitrogen, Camarillo, CA, USA</td><td>KRC0011</td><td></td></tr><tr><td>IL-6 Rat ELISA Kit</td><td>Invitrogen, Camarillo, CA, USA</td><td>KRC0061</td><td></td></tr><tr><td>INF-gamma Rat ELISA Kit</td><td>Invitrogen, Camarillo, CA, USA</td><td>KRC4021</td><td></td></tr><tr><td>TNF-alpha Rat ELISA Kit</td><td>Invitrogen, Camarillo, CA, USA</td><td>KRC3011</td><td></td></tr><tr><td>Rat LPS ELISA Kit</td><td>NeoBioLab, MA, USA</td><td>RL0275</td><td></td></tr><tr><td>Environmental Chamber 6020-1</td><td>Caron, Marietta, OH, USA</td><td>6020-1</td><td></td></tr><tr><td>Centrifuge&amp;nbsp;</td><td>Beckman Coulter, Indianapolis, IN, USA</td><td>Optima L-90K</td><td></td></tr><tr><td>Ultra Centrifuge</td><td></td><td></td><td></td></tr><tr><td>Light microscope optical system</td><td>CitoViva Technology Inc., Auburn, AL</td><td></td><td></td></tr><tr><td>Colony counter</td><td>Fisher Scientific , Pittsburgh, PA, USA</td><td>RE-3325</td><td></td></tr><tr><td>Freezer</td><td>Haier, Brooklyn, NY, USA</td><td>HCMO50LA</td><td></td></tr><tr><td>Ultra microplate reader</td><td>Bio-Tek Instrument, Winooski, VT, USA</td><td>ELx 808</td><td></td></tr><tr><td>Auto Strip Washer</td><td>Bio-Tek Instrument, Winooski, VT, USA</td><td>ELx50</td><td></td></tr><tr><td>Pipettes</td><td>Gilson, Pipetman, France</td><td>P100, P200, P1000</td><td></td></tr><tr><td>C24 Incubator Shaker</td><td>New Brunswick Scientific, Enfield, CT, USA</td><td>Classic C24</td><td></td></tr><tr><td>Rocking Shaker&amp;nbsp;</td><td>Reliable Scientific, Inc., Nesbit, MS, USA</td><td>55</td><td></td></tr><tr><td>Petri dishes</td><td>Fisher Scientific, Pittsburgh, PA, USA</td><td>875713</td><td>100 mm x 15 mm</td></tr><tr><td>SterilGard III Advance</td><td>The Baker Company, Sanford, ME, USA</td><td>SG403</td><td></td></tr><tr><td>Culture Growing Flasks</td><td>Corning Incorporated, Corning, NY, USA</td><td>4995</td><td>PYREX 250 ml Erlenmeyer flasks</td></tr><tr><td>Optical Spectrometer Genesys 20</td><td>Thermo Scientific, Waltham, MA, USA.</td><td>4001</td><td></td></tr><tr><td>Sony DXC-33 Video camera</td><td>Sony</td><td></td><td></td></tr><tr><td>Richardson test slide</td><td>Electron Microscopy Science, Hatfield, PA, USA</td><td>80303</td><td></td></tr><tr><td>Millipore water purification system</td><td>Millipore</td><td>Direct-Q</td><td></td></tr><tr><td>Image-Pro Plus software</td><td>Media Cybernetics, MD, USA</td><td></td><td></td></tr><tr><td>Triple Beam Balance</td><td>OHAUS Corporation, Parsippany, NJ, USA</td><td></td><td></td></tr><tr><td>Tissue Homogenizer, Dounce</td><td>VWR</td><td>71000-518</td><td></td></tr></tbody></table>

prevention of heat stress adverse effects in rats by bacillus subtilis strain

Diese Studie wurde entworfen , um die Schutzwirkung des Bacillus subtilis Stamms gegen Komplikationen Stress aufzuheizen Bezug zu bewerten. Zweiunddreißig Sprague-Dawley-Ratten wurden in dieser Studie verwendet. Die Tiere wurden oral zweimal täglich für zwei Tage mit B behandelt subtilis - Stamm BSB3 oder PBS. Am nächsten Tag nach der letzten Behandlung wurde jede Gruppe eingeteilt und zwei experimentelle Gruppen (eine mit PBS behandelt und eine mit B. subtilis - behandelt) wurden für 25 min bei 45 o C gebracht. Zwei Kontrollgruppen waren für 25 Minuten bei Raumtemperatur. Alle Ratten wurden eingeschläfert und verschiedene Parameter wurden in allen Gruppen analysiert. Unerwünschte Wirkungen von Hitzestress werden durch die Abnahme der Zotten Höhe und Gesamt Mukosadicke im Darmepithel registriert; Translokation von Bakterien aus dem Lumen; Vesikulation von Erythrozyten und Erhebung der Lipopolysaccharide (LPS) im Blut erhöht. Die schützende Wirksamkeit der Behandlung wird durch pre ausgewertetErfindung dieser Nebenwirkungen. Das Protokoll wurde für die orale Behandlung von Ratten, die mit Bakterien zur Verhinderung von Hitzestress Komplikationen eingerichtet, aber dieses Protokoll kann auch für andere Verabreichungswege und für die Analyse verschiedener Verbindungen modifiziert und verwendet werden.

Die mittlere Körpertemperatur der Tiere vor und unmittelbar nach der Hitzestress war 36,7 ± 0,07 ° C und 40,3 ± 0,17 ° C, bzw. (P &lt;0,05). Exposition von Ratten gegenüber Wärme (Gruppe 3) führte zu einer signifikanten Hemmung der Zotten Höhe und Gesamtschleimhautdicke (2, 3). Die orale Verabreichung von BSB3 Stamm vor Stress geschützt Darm vor der schädlichen Wirkung von Wärme. <p class="jove_content" fo:keep-together.within-page="...

Watch this Scientific Journal Video about Prevention of Heat Stress Adverse Effects in Rats by Bacillus subtilis Strain at JoVE.com

Prevention of Heat Stress Adverse Effects in Rats by Bacillus subtilis Strain

We described a protocol for prevention of heat stress effects in rats by oral pre-treatment with beneficial bacteria. This protocol can be modified and used for various routes of administration and for analysis of different compounds.

Prävention von Hitzestress Unerwünschte Wirkungen bei Ratten durch<em&gt; Bacillus subtilis</em&gt; Dehnungs

This study was designed to evaluate the protective effect of the Bacillus subtilis strain against complications related to heat stress. Thirty-two ...

prevention-heat-stress-adverse-effects-rats-bacillus-subtilis

Research

JoVE Journal

Immunology and Infection

Prevention of Heat Stress Adverse Effects in Rats by Bacillus subtilis Strain

7.4K Aufrufe.  Auburn University. We described a protocol for prevention of heat stress effects in rats by oral pre-treatment with beneficial bacteria. This protocol can be modified and used for various routes of administration and for analysis of different compounds.

Serie: Prevention of Heat Stress Adverse Effects in Rats by Bacillus subtilis Strain

A Rat Methyl-Seq Platform to Identify Epigenetic Changes Associated with Stress Exposure

As genomes of a wider variety of animals become available, there is an increasing need for tools that can capture dynamic epigenetic changes in these animal models. The rat is one particular model animal where an epigenetic tool can complement many pharmacological and behavioral studies to provide insightful mechanistic information. To this end, we adapted the SureSelect Target Capture System (referred to as Methyl-Seq) for the rat, which can assess DNA methylation levels across the rat genome. The rat design targeted promoters, CpG islands, island shores, and GC-rich regions from all RefSeq genes. 
To implement the platform on a rat experiment, male Sprague Dawley rats were exposed to chronic variable stress for 3 weeks, after which blood samples were collected for genomic DNA extraction. Methyl-Seq libraries were constructed from the rat DNA samples by shearing, adapter ligation, target enrichment, bisulfite conversion, and multiplexing. Libraries were sequenced on a next-generation sequencing platform and the sequenced reads were analyzed to identify DMRs between DNA of stressed and unstressed rats. Top candidate DMRs were independently validated by bisulfite pyrosequencing to confirm the robustness of the platform.
Results demonstrate that the rat Methyl-Seq platform is a useful epigenetic tool that can capture methylation changes induced by exposure to stress.

Here, we describe the protocol and implementation of Methyl-Seq, an epigenomic platform, using a rat model to identify epigenetic changes associated with chronic stress exposure. Results demonstrate that the rat Methyl-Seq platform is capable of detecting methylation differences that arise from stress exposure in rats.

Eine Ratte Methyl-Seq-Plattform um epigenetische Veränderungen im Zusammenhang mit Stress Exposition zu identifizieren

As genomes of a wider variety of animals become available, there is an increasing need for tools that can capture dynamic epigenetic changes in these ...

Forschung

Biochemie

Standardized Methods for Measuring Induction of the Heat Shock Response in Caenorhabditis elegans

The heat shock response (HSR) is a cellular stress response induced by cytosolic protein misfolding that functions to restore protein folding homeostasis, or proteostasis. Caenorhabditis elegans occupies a unique and powerful niche for HSR research because the HSR can be assessed at the molecular, cellular, and organismal levels. Therefore, changes at the molecular level can be visualized at the cellular level and their impacts on physiology can be quantitated at the organismal level. While assays for measuring the HSR are straightforward, variations in the timing, temperature, and methodology described in the literature make it challenging to compare results across studies. Furthermore, these issues act as a barrier for anyone seeking to incorporate HSR analysis into their research. Here, a series of protocols is&#160;presented for measuring induction of the HSR in a robust and reproducible manner with RT-qPCR, fluorescent reporters, and an organismal thermorecovery assay. Additionally, we show that a widely used thermotolerance assay is not dependent on the well-established master regulator of the HSR, HSF-1, and therefore should not be used for HSR research. Finally, variations in these assays found in the literature are discussed and best practices are proposed to help standardize results across the field, ultimately facilitating neurodegenerative disease, aging, and HSR research.

Standardized Methods for Measuring Induction of the Heat Shock Response in Caenorhabditis elegans

Here, standardized protocols are presented to assess induction of the heat shock response (HSR) in Caenorhabditis elegans using RT-qPCR at the molecular level, fluorescent reporters at the cellular level, and thermorecovery at the organismal level.

Standardisierte Methoden zur Messung der Induktion der Hitzeschockreaktion bei Caenorhabditis elegans

The heat shock response (HSR) is a cellular stress response induced by cytosolic protein misfolding that functions to restore protein folding homeostasis, ...

Umwelt

Isolation, Purification, and Identification of Bacitracin-producing Bacillus licheniformis from Fresh Feces of Healthy Pigs

Bacillus licheniformis and bacitracin have a huge application market and value in the fields of medicine, chemistry, aquaculture, agricultural, and sideline products. Therefore, the selection of B. licheniformis with high production of bacitracin is of great importance. In this experimental protocol, Bacillus with a high yield of bacitracin was isolated, purified, and identified from the fresh feces of healthy pigs. The inhibitory effect of secondary metabolite bacitracin on Micrococcus luteus was also tested. Thin-layer chromatography and high-performance liquid chromatography were used for the qualitative and quantitative detection of bacitracin. The physiological and biochemical characteristics of B. licheniformis were determined by relevant kits. The phylogenetic relationships of B. licheniformis were determined and constructed using gene sequence detection. This protocol describes and introduces the standard isolation, purification, and identification process of B. licheniformis from animal fresh feces from multiple perspectives, providing a method for the large-scale utilization of B. licheniformis and bacitracin in factories.

Isolation, Purification, and Identification of Bacitracin-producing Bacillus licheniformis from Fresh Feces of Healthy Pigs

This protocol provides a complete and comprehensive description of the detailed process for the isolation, purification, and identification of bacitracin-producing Bacillus licheniformis from healthy pig feces.

Bacillus licheniformis and bacitracin have a huge application market and value in the fields of medicine, chemistry, aquaculture, agricultural, and ...

Biotechnik

Short-Duration Hypothermia Induction in Rats using Models for Studies examining Clinical Relevance and Mechanisms

Therapeutic hypothermia (TH) is a powerful neuroprotective strategy that has provided robust evidence for neuroprotection in pre-clinical studies of neurological disorders. Despite strong pre-clinical evidence, TH has not shown efficacy in clinical trials of most neurological disorders. The only successful trials employing therapeutic hypothermia were related to cardiac arrest in adults and hypoxic ischemic injury in neonates. Further investigations into the parameters of its use, and study design comparisons between pre-clinical and clinical studies, are warranted. This article demonstrates two methods of short-duration hypothermia induction. The first method allows for rapid hypothermia induction in rats using ethanol spray and fans. This method works by cooling the skin, which has been less commonly used in clinical trials and may have different physiological effects. Cooling is much more rapid with this technique than is achievable in human patients due to differences in surface area to volume ratio. Along with this, a second method is also presented, which allows for a clinically achievable cooling rate for short-duration hypothermia. This method is easy to implement, reproducible and does not require active skin cooling.

This article describes two methods of whole-body short-duration hypothermia induction in rats. The first, rapid induction method, employs active cooling using fans and ethanol spray for a rapid decrease in temperature. The second method is a gradual cooling method. This is achieved using the combination of isoflurane anesthesia and the reduction of temperature settings on the homeothermic heat mat. This results in a gradual decrease in core body temperature without the use of any external cooling devices.&#160;

Kurzzeit-Hypothermie-Induktion bei Ratten unter Verwendung von Modellen für Studien zur Untersuchung klinischer Relevanz und Mechanismen

Therapeutic hypothermia (TH) is a powerful neuroprotective strategy that has provided robust evidence for neuroprotection in pre-clinical studies of ...

Medizin

A Preclinical Model of Exertional Heat Stroke in Mice

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, whereas exertional heat stroke (EHS) occurs during physical activity. EHS differs from CHS in etiology, clinical presentation, and sequelae of multi-organ dysfunction. Until recently, only models of CHS have been well established. This protocol aims to provide guidelines for a refined preclinical mouse model of EHS that is free from major limiting factors such as the use of anesthesia, restraint, rectal probes, or electric shock. Male and female C57Bl/6 mice, instrumented with core temperature (Tc) telemetric probes were utilized in this model. For familiarization with the running mode, mice undergo 3 weeks of training using both voluntary and forced running wheels. Thereafter, mice run on a forced wheel inside a climatic chamber set at 37.5 &#176;C and 40%-50% relative humidity (RH) until displaying symptom limitation (e.g., loss of consciousness) at Tc of 42.1-42.5 &#176;C, although suitable results can be obtained at chamber temperatures between 34.5-39.5 &#176;C and humidity between 30%-90%. Depending on the desired severity, mice are removed from the chamber immediately for recovery in ambient temperature or remain in the heated chamber for a longer duration, inducing a more severe exposure and a higher incidence of mortality. Results are compared with sham-matched exercise controls (EXC) and/or na&#239;ve controls (NC). The model mirrors many of the pathophysiological outcomes observed in human EHS, including loss of consciousness, severe hyperthermia, multi-organ damage as well as inflammatory cytokine release, and acute phase responses of the immune system. This model is ideal for hypothesis-driven research to test preventative and therapeutic strategies that may delay the onset of EHS or reduce the multi-organ damage that characterizes this manifestation.

The protocol describes the development of a standardized, repeatable, preclinical model of exertional heat stroke (EHS) in mice free from adverse external stimuli such as electric shock. The model provides a platform for mechanistic, preventative, and therapeutic studies.

Ein präklinisches Modell des Belastungshitzeinschlags bei Mäusen

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, ...

Investigating the Detrimental Effects of Low Pressure Plasma Sterilization on the Survival of Bacillus subtilis Spores Using Live Cell Microscopy

Plasma sterilization is a promising alternative to conventional sterilization methods for industrial, clinical, and spaceflight purposes. Low pressure plasma (LPP) discharges contain a broad spectrum of active species, which lead to rapid microbial inactivation. To study the efficiency and mechanisms of sterilization by LPP, we use spores of the test organism Bacillus subtilis because of their extraordinary resistance against conventional sterilization procedures. We describe the production of B. subtilis spore monolayers, the sterilization process by low pressure plasma in a double inductively coupled plasma reactor, the characterization of spore morphology using scanning electron microscopy (SEM), and the analysis of germination and outgrowth of spores by live cell microscopy. A major target of plasma species is genomic material (DNA) and repair of plasma-induced DNA lesions upon spore revival is crucial for survival of the organism. Here, we study the germination capacity of spores and the role of DNA repair during spore germination and outgrowth after treatment with LPP by tracking fluorescently-labelled DNA repair proteins (RecA) with time-resolved confocal fluorescence microscopy. Treated and untreated spore monolayers are activated for germination and visualized with an inverted confocal live cell microscope over time to follow the reaction of individual spores. Our observations reveal that the fraction of germinating and outgrowing spores is dependent on the duration of LPP-treatment reaching a minimum after 120 s. RecA-YFP (yellow fluorescence protein) fluorescence was detected only in few spores and developed in all outgrowing cells with a slight elevation in LPP-treated spores. Moreover, some of the vegetative bacteria derived from LPP-treated spores showed an increase in cytoplasm and tended to lyse. The described methods for analysis of individual spores could be exemplary for the study of other aspects of spore germination and outgrowth.

Investigating the Detrimental Effects of Low Pressure Plasma Sterilization on the Survival of Bacillus subtilis Spores Using Live Cell Microscopy

This protocol illustrates the important consecutive steps required to assess the relevance of monitoring vitality parameter and DNA repair processes in reviving Bacillus subtilis spores after treatment with low pressure plasma by tracking fluorescence-labelled DNA repair proteins via time-resolved confocal microscopy and scanning electron microscopy.

Untersucht die nachteiligen Effekte der Low Druck Plasmasterilisation auf das Überleben von Bacillus Subtilis Sporen mittels Live Cell Mikroskopie

Plasma sterilization is a promising alternative to conventional sterilization methods for industrial, clinical, and spaceflight purposes. Low pressure ...

Biologie

Etablierung und Validierung eines Modells für chronisch unvorhersehbaren leichten Stress

Chronic Unpredictable Mild Stress in Rats based on the Mongolian medicine

Depression is a prevalent affective disorder and constitutes a leading cause of global disability. The limitations of current pharmacological interventions contribute to the substantial health burden attributed to this condition. There is a pressing need for a deeper understanding of the underlying mechanisms of depression, making pre-clinical models with translational potential highly valuable. Mongolian medicine, a subset of traditional medicine, posits that disease occurrence is closely tied to the equilibrium of wind, bile, and Phlegm. In this study, we introduce a protocol for the chronic unpredictable mild stress (CUMS) method in rats. Within this framework, rats are subjected to a series of fluctuating, mild stressors to induce a depression-like phenotype, mimicking the pathogenesis of human depression. Behavioral assays employed in this protocol include the sucrose preference test (SPT), indicative of anhedonia-a core symptom of depression; the open field test (OFT), which measures anxiety levels; and the Morris water maze test (MWM), which evaluates spatial memory and learning abilities. The CUMS method demonstrates the capability to induce anhedonia and to cause long-term behavioral deficits. Furthermore, this protocol is more aligned with Mongolian medical theory than other animal models designed to elicit depression-like behavior. The development of this animal model and subsequent research provide a robust foundation for future innovative studies in the realm of Mongolian medicine.

Autor im Rampenlicht: Etablierung eines Nagetiermodells zur Untersuchung von Depressionsfaktoren in der traditionellen mongolischen Medizin

This protocol outlines a chronic unpredictable mild stress (CUMS) model for depression based on Mongolian medical theory, along with methods for validating behavioral tests.

Chronisch unvorhersehbarer milder Stress bei Ratten basierend auf der mongolischen Medizin

Depression is a prevalent affective disorder and constitutes a leading cause of global disability. The limitations of current pharmacological ...

Measurements of Physiological Stress Responses in C. Elegans

Organisms are often exposed to fluctuating environments and changes in intracellular homeostasis, which can have detrimental effects on their proteome and physiology. Thus, organisms have evolved targeted and specific stress responses dedicated to repair damage and maintain homeostasis. These mechanisms include the unfolded protein response of the endoplasmic reticulum (UPRER), the unfolded protein response of the mitochondria (UPRMT), the heat shock response (HSR), and the oxidative stress response (OxSR). The protocols presented here describe methods to detect and characterize the activation of these pathways and their physiological consequences in the nematode, C. elegans. First, the use of pathway-specific fluorescent transcriptional reporters is described for rapid cellular characterization, drug screening, or large-scale genetic screening (e.g., RNAi or mutant libraries). In addition, complementary, robust physiological assays are described, which can be used to directly assess sensitivity of animals to specific stressors, serving as functional validation of the transcriptional reporters. Together, these methods allow for rapid characterization of the cellular and physiological effects of internal and external proteotoxic perturbations.

Measurements of Physiological Stress Responses in C. Elegans

Here, we characterize cellular proteotoxic stress responses in the nematode C. elegans by measuring the activation of fluorescent transcriptional reporters and assaying sensitivity to physiological stress.

Messung physiologischer Stressreaktionen in C. Elegans

Organisms are often exposed to fluctuating environments and changes in intracellular homeostasis, which can have detrimental effects on their proteome and ...

Prävention von Hitzestress Unerwünschte Wirkungen bei Ratten durch

Zusammenfassung

Weitere Videos entdecken

Kapitel in diesem Video

Eine Ratte Methyl-Seq-Plattform um epigenetische Veränderungen im Zusammenhang mit Stress Exposition zu identifizieren

Standardisierte Methoden zur Messung der Induktion der Hitzeschockreaktion bei Caenorhabditis elegans

Isolation, Purification, and Identification of Bacitracin-producing Bacillus licheniformis from Fresh Feces of Healthy Pigs

Kurzzeit-Hypothermie-Induktion bei Ratten unter Verwendung von Modellen für Studien zur Untersuchung klinischer Relevanz und Mechanismen

Ein präklinisches Modell des Belastungshitzeinschlags bei Mäusen

Untersucht die nachteiligen Effekte der Low Druck Plasmasterilisation auf das Überleben von Bacillus Subtilis Sporen mittels Live Cell Mikroskopie

Chronisch unvorhersehbarer milder Stress bei Ratten basierend auf der mongolischen Medizin

Chronisch unvorhersehbarer milder Stress bei Ratten basierend auf der mongolischen Medizin

Chronisch unvorhersehbarer milder Stress bei Ratten basierend auf der mongolischen Medizin

Messung physiologischer Stressreaktionen in C. Elegans