Name: analysis of the epithelial damage produced by entamoeba histolytica infection
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This work was supported by grants from the Institute of Science and Technology of the Federal District (ICyTDF, 64/2012 to EO) and the Mexican Council for Science and Technology (Conacyt, 179895 to MS).

1. Opzetten en onderhouden van E. histolytica Cultures <ol><li> Grow axenisch (geheel vrij van alle andere verontreinigende organismen) 1 x 10 5 trofozoiten van Entamoeba histolytica stam HML: IMSS klonen A 18 in 16 x 125 mm cultuur buizen met Teflon liner schroefdoppen (of 1 x 10 6 trofozoïeten in een wegwerp T- 25 kolf) en 15 ml (of 50 ml in T-25 kolf) van TYI-S-33 medium (TYI bouillon aangevuld met 3% Diamond vitaminemengsel, 10% hitte geïnactiveerd volwassen run...

<ol>
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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cysteine+proteases+of+parasitic+organisms.">Cysteine proteases of parasitic organisms.</a> Mol Biochem Parasitol. 120, 1-21 (2002).</li><li>Betanzos, 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=The+EhCPADH112+complex+of+Entamoeba+histolytica+interacts+with+tight+junction+proteins+occludin+and+claudin-1+to+produce+epithelial+damage.">The EhCPADH112 complex of Entamoeba histolytica interacts with tight junction proteins occludin and claudin-1 to produce epithelial damage.</a> PLoS One. 8, (2013).</li><li>Lauwaet, T., 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=Proteinase+inhibitors+TPCK+and+TLCK+prevent+Entamoeba+histolytica+induced+disturbance+of+tight+junctions+and+microvilli+in+enteric+cell+layers+in+vitro.">Proteinase inhibitors TPCK and TLCK prevent Entamoeba histolytica induced disturbance of tight junctions and microvilli in enteric cell layers in vitro.</a> Int J Parasitol. 34, 785-794 (2004).</li><li>Leroy, 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=Contact-dependent+transfer+of+the+galactose-specific+lectin+of+Entamoeba+histolytica+to+the+lateral+surface+of+enterocytes+in+culture.">Contact-dependent transfer of the galactose-specific lectin of Entamoeba histolytica to the lateral surface of enterocytes in culture.</a> Infect Immun. 63, 4253-4260 (1995).</li><li>Leroy, A., Lauwaet, T., De Bruyne, G., Cornelissen, M., Mareel, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Entamoeba+histolytica+disturbs+the+tight+junction+complex+in+human+enteric+T84+cell+layers.">Entamoeba histolytica disturbs the tight junction complex in human enteric T84 cell layers.</a> FASEB J. 14, 1139-1146 (2000).</li><li>Leroy, 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=Disturbance+of+tight+junctions+by+Entamoeba+histolytica:+resistant+vertebrate+cell+types+and+incompetent+trophozoites.">Disturbance of tight junctions by Entamoeba histolytica: resistant vertebrate cell types and incompetent trophozoites.</a> Arch Med Res. 31, (2000).</li><li>Lauwaet, T., 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=Entamoeba+histolytica+trophozoites+transfer+lipophosphopeptidoglycans+to+enteric+cell+layers.">Entamoeba histolytica trophozoites transfer lipophosphopeptidoglycans to enteric cell layers.</a> Int J Parasitol. 34, 549-556 (2004).</li><li>Kissoon-Singh, V., Moreau, F., Trusevych, E., Chadee, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Entamoeba+histolytica+Exacerbates+Epithelial+Tight+Junction+Permeability+and+Proinflammatory+Responses+in+Muc2(-/-)+Mice.">Entamoeba histolytica Exacerbates Epithelial Tight Junction Permeability and Proinflammatory Responses in Muc2(-/-) Mice.</a> Am J Pathol. 182, 852-865 (2013).</li><li>Lejeune, M., Moreau, F., Chadee, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prostaglandin+E2+produced+by+Entamoeba+histolytica+signals+via+EP4+receptor+and+alters+claudin-4+to+increase+ion+permeability+of+tight+junctions.">Prostaglandin E2 produced by Entamoeba histolytica signals via EP4 receptor and alters claudin-4 to increase ion permeability of tight junctions.</a> Am J Pathol. 179, 807-818 (2011).</li><li>Orozco, M. 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Om in vitro gastheer-pathogeen interacties tijdens epitheliale infectie te bestuderen door E. histolytica, is het cruciaal om te werken met gevestigde culturen van zowel epitheelcellen en trofozoiten. Bijvoorbeeld, vroeger, E. histolytica culturen had meestal in combinatie met bepaalde soorten bacteriën of trypanosomatids 22,23 vastgesteld. Echter, co-kweken van E. histolytica culturen contraproductief voor de studie van gastheer-pathogeen interacties omdat waargenomen eff...

Entamoeba histolytica is een eencellige protozoaire verantwoordelijk van menselijke amoebiasis, een darminfectie veroorzaakt ontsteking en diarree. E. histolytica infecteert tot 50 miljoen mensen per jaar, maar slechts ongeveer 10% van de geïnfecteerde mensen ontwikkelen de symptomen verbonden met amoebiasis 1. Infectie optreedt na inname van besmet voedsel of water met E. histolytica cysten. In de darm, cysten produceren levende trofozoïeten die voldoen aan dikke darm mucine en vermenigvuldigen 2. Trofozoiten vormen meestal cysten die worden uitgescheiden via de ontlasting. In andere gevallen en voor nog onbekende redenen, trofozoïeten breken de intestinale epitheliale laag en binnenvallen onderliggende weefsels. In het ergste geval, ze in de bloedbaan en invloed op andere organen zoals de lever 3. Het doorbreken van de epitheliale barrière vereist verstoring van epitheliale transmembraanpenetratie structuren die cellen toegetreden handhaven. Epitheelcellencontacten worden gevormd door de apicale junctionele complex bestaande uit vast (TJ) en contactplaatsen (AJ) en desmosomen 4. De meest apicale knooppunten zijn TJ, en daarom zijn ze de eerste barrière beledigd door E. histolytica en enkele andere ziekteverwekkers tijdens gastheer invasie. TJ bestaan ​​uit transmembraanpenetratie adhesie receptoren zoals Claudins, occludin en junctionele adhesiemoleculen (JAM) die zich bezighouden met homo-of heterofiele interacties met receptoren van de naburige cel. Ze worden intracellulair gebonden aan steiger moleculen van de zonula occludens (ZO) familie die adhesiereceptoren aansluiten op actinecytoskelet verdere mechanische sterkte aan het epitheel. TJ zijn verantwoordelijk voor het afdichten van darmweefsel uit de darm lumen, het voorkomen van overmatig water en opgeloste stof lekken. Zo, na TJ worden verstoord door de parasiet, weefsels zijn binnengevallen. E. histolytica scheidt verschillende moleculen, zoals: (i) die betrokken is bij hechting van amoebae te target 5 cellen; (Ii) membraan-actieve factoren die aan doden van gastheercellen door exocytose, bijvoorbeeld ion-channel vormende peptiden genoemd amoebapores 6,7; en (iii) proteïnasen die extracellulaire matrix afbreken en bemiddelen weefsel desintegratie 5,8,9. De cysteïne protease EhCP112 en adhesiemoleculen EhADH112 die samen de EhCPADH112 complex twee E. histolytica virulentie eiwitten die een belangrijke rol spelen bij de demontage van TJ 10. Levende trofozoiten, hun totale lysaten en uitgescheiden te induceren moleculaire veranderingen in de TJ complexe en functionele verstoring van de epitheliale barrière. In deze studie wordt aangetoond dat EhCP112 en EhADH112 interactie met occludin en claudine-1 eiwitten leidt tot internalisatie en degradatie van cel eiwitten, waardoor E. vergemakkelijkt histolytica entree via de paracellulaire pad. Onze gegevens en die of andere groepen 11-17 suggereren sterk de noodzaak van specifieke gastheer-pathogeen interacties die parasiet invasie mogelijk te maken. Het ontrafelen van de moleculaire basis van deze interacties is van groot belang voor een beter begrip van amoebiasis pathogenese. Selectieve verstoring van TJ door trofozoïeten, gekenmerkt door verhoogde paracellulaire permeabiliteit kan worden gemeten door een afname in transepitheliale elektrische weerstand (TER). De overdracht van parasitaire eiwitten naar gastheer epitheel kan worden bepaald door immunofluorescentie kleuring en confocale laser microscopie, een werkwijze die ook co-lokalisatie van amoebe virulentiefactoren epitheliale junctions bebakening mogelijke directe interacties kunnen onthullen. In dit artikel beschrijven we in detail hoe epitheelcellen en trofozoiten worden gekweekt, geoogst en gemanipuleerd om gastheer-pathogeen interacties en de gevolgen daarvan te onderzoeken.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Entamoeba histolytica HM1:IMSS, Clone A&#160;</td>
			<td>IMSS Hospital, Mexico</td>
			<td>Without/number</td>
			<td>Virulent trophozoites18&#160;</td>
		</tr>
		<tr>
			<td rowspan="8">TYI broth</td>
			<td rowspan="8">Becton, Dickinson and Company 
			Merck 
			Merck 
			Merck 
			J.T. Baker 
			Reproquifin 
			SIGMA-Aldrich 
			SIGMA-Aldrich</td>
			<td rowspan="8">211862 
			K35625437 626 
			21578 
			4873 
			3252-01 
			CAS 50-81-7 
			C7880 
			F-5879</td>
			<td rowspan="8">3.45% BBL Biosate peptone 
			58 mM glucose 
			39 mM NaCl 
			5 mM KH2PO4 
			6.5 mM K2HPO4 
			16.3 mM ascorbic acid 
			8.1 mM L-cysteine 
			&#160;0.1 mM ferric ammonium citrate, adjust pH 6.819</td>
		</tr>
		<tr>
			<td>Bovine serum adult</td>
			<td>Microlab , Labs., Mex.</td>
			<td>SU146</td>
			<td>Use at 10% and inactivated to 56&#176; C for 30 min</td>
		</tr>
		<tr>
			<td>Diamond&#160; vitamin&#160; mixture- Tween 80</td>
			<td>In vitro</td>
			<td>SR-07</td>
			<td>Use at 3%</td>
		</tr>
		<tr>
			<td>Penicillin&#160;</td>
			<td>Lakeside,&#160; M&#233;x.</td>
			<td>34564SSA IV</td>
			<td>0.5 IU/mL</td>
		</tr>
		<tr>
			<td>Streptomycin&#160;</td>
			<td>Lakeside,&#160; M&#233;x.</td>
			<td>75757SSA IV</td>
			<td>35 &#181;g/ mL</td>
		</tr>
		<tr>
			<td>Pyrex 15 mL screw cap culture tubes with PTFE lined phenolic caps</td>
			<td>Corning-Pyrex</td>
			<td>9826-16X</td>
			<td>16x125 mm, capacity 15 mL and caps fabricated from special formula resistant to effects of temperature</td>
		</tr>
		<tr>
			<td>Cell culture plates, 6 Well</td>
			<td>Corning-Costar</td>
			<td>3516</td>
			<td>Sterile plates, well diameter 34.8 mm and growth area 9.5 cm2.&#160; Rings on lid prevent cross-contamination</td>
		</tr>
		<tr>
			<td>25cm2 cell culture flask</td>
			<td>Corning-Costar</td>
			<td>430168</td>
			<td>Canted neck flasks</td>
		</tr>
		<tr>
			<td>MDCK (Madin Darby canine kidney) type I</td>
			<td>American Type Culture Collection</td>
			<td>CCL34</td>
			<td>Kidney epithelial cells grown between the 60th and 90th passage</td>
		</tr>
		<tr>
			<td>DMEM medium&#160;</td>
			<td>Gibco&#160;</td>
			<td>12800-017</td>
			<td>Dulbecco&#39;s Modified Eagle Medium with high glucose.</td>
		</tr>
		<tr>
			<td>Neonate Calf Serum</td>
			<td>In vitro</td>
			<td>S-02</td>
			<td>Use at 10%.&#160;&#160;</td>
		</tr>
		<tr>
			<td>Penicillin/Streptomycin mixture&#160;</td>
			<td>In vitro&#160;</td>
			<td>A-01</td>
			<td>Stock solution 10,000 U/&#181;g/mL</td>
		</tr>
		<tr>
			<td>Insulin&#160;&#160;</td>
			<td>AMSA</td>
			<td>398MJ94SSA IV</td>
			<td>Stock solution 100 IU/mL</td>
		</tr>
		<tr>
			<td>Trypsin solution&#160;</td>
			<td>In vitro</td>
			<td>EN-005</td>
			<td>0.05% enzyme solution without calcium and magnesium</td>
		</tr>
		<tr>
			<td>75cm2 cell culture flask</td>
			<td>Corning-Costar</td>
			<td>430720</td>
			<td>Canted neck flasks for trophozoite culture in TYI-S-33 medium</td>
		</tr>
		<tr>
			<td>Transwell permeable supports</td>
			<td>Corning-Costar</td>
			<td>3470</td>
			<td>0.4. &#181;m polyster membrane, 6.5 mm insert in 24 well plate, growth area 0.3 cm2</td>
		</tr>
		<tr>
			<td>24 well cell culture dish&#160;&#160;</td>
			<td>Corning-Costar</td>
			<td>3524</td>
			<td>Clear polystyrene, treated for optimal cell attachment, sterilized by gamma radiation and certified non-pyrogenic</td>
		</tr>
		<tr>
			<td>Complete Mini</td>
			<td>Roche</td>
			<td>11836 153 001</td>
			<td>Protease inhibitor cocktail inhibits a broad spectrum of serine, cysteine and metallo-proteases. Final concentration 1 mM&#160;</td>
		</tr>
		<tr>
			<td>Trans-epoxysuccinyl-L-leucylamido (4-guanidino) butane (E-64)</td>
			<td>SIGMA-Aldrich</td>
			<td>E3132</td>
			<td>Cystein protease inhibitor, final concentration 40 &#181;g/mL</td>
		</tr>
		<tr>
			<td>p&#945;ZO-1&#160;</td>
			<td>Invitrogen</td>
			<td>402200</td>
			<td>IgG rabbit policlonal&#160; antibody&#160; against&#160; a synthetic peptide in the middle region of the ZO-1 human protein</td>
		</tr>
		<tr>
			<td>m&#945;EhCPADH112</td>
			<td>Homemade antibody</td>
			<td>Without/ Number</td>
			<td>IgM mouse monoclonal antibody&#160; against&#160; 444-601 epitope located at C-terminal of EhCPADH11221,27</td>
		</tr>
		<tr>
			<td>FITC-goat anti-mouse IgM</td>
			<td>Zymed</td>
			<td>62-6811</td>
			<td>Fluorescein isotiocyanate (FITC)-labelled goat anti-mouse secondary&#160; antibody</td>
		</tr>
		<tr>
			<td>TRITC- goat anti-rabbit IgG (H+L)</td>
			<td>Zymed</td>
			<td>816114</td>
			<td>Tetramethyl-rhodamine isothiocyanate (TRITC)-labelled&#160; goat anti-rabbit IgG&#160; secondary antibody.</td>
		</tr>
		<tr>
			<td>STX2 Electrode</td>
			<td>World Precision Instrument&#160;</td>
			<td>102711</td>
			<td>Consists of a fixed pair of double electrodes, 4 mm wide and 1 mm thick. Each stick of the electrode pair contains a silver/silver-chloride pellet for measuring voltage and a silver electrode for passing current. For use with EVOM</td>
		</tr>
		<tr>
			<td>EVOM epithelial voltohmmeter</td>
			<td>World Precision Instrument&#160;</td>
			<td>12111</td>
			<td>Use in resistance mode and maintain unplugged during TER measurements</td>
		</tr>
		<tr>
			<td>Neubauer chamber</td>
			<td>MEARIENFELD</td>
			<td>610610</td>
			<td>Hemocytometer&#160;</td>
		</tr>
		<tr>
			<td>Leica TCS_SP5_MO</td>
			<td>Leica</td>
			<td>Without/number</td>
			<td>Laser confocal microscopy with Leica microsystems CMS Gmbh/leica Las af Lite/BIN software</td>
		</tr>
		<tr>
			<td>Vectashield</td>
			<td>Vector Laboratories, Inc.</td>
			<td>H-1000</td>
			<td>Mounting medium for fluorescence</td>
		</tr>
		<tr>
			<td>4&#180;, 6-diamino-2-phenylindole (Dapi)</td>
			<td>SIGMA</td>
			<td>D-9542</td>
			<td>0.05 mM final concentration</td>
		</tr>
		<tr>
			<td>Bovine serum albumin (BSA)</td>
			<td>US Biological</td>
			<td>A-1310</td>
			<td>0.5%&#160; final concentration for blocking solution</td>
		</tr>
	</tbody>
</table>

analysis of the epithelial damage produced by entamoeba histolytica infection

Entamoeba histolytica is de verwekker van de menselijke amoebiasis, een belangrijke oorzaak van diarree en lever abces in tropische landen. Infectie wordt geïnitieerd door interactie van het pathogeen met intestinale epitheelcellen. Deze interactie leidt tot verstoring van intercellulaire structuren zoals tight junctions (TJ). TJ zorgen voor afdichting van de epitheliale laag om gastweefsel scheiden van darmlumen. Recente studies tonen aan dat verstoring van TJ door de parasitaire eiwit EhCPADH112 is een voorwaarde voor E. histolytica invasie die gepaard gaat met epitheliale barrière disfunctie. De analyse van de moleculaire mechanismen betrokken bij TJ demontage tijdens E. histolytica invasie is van het grootste belang voor ons begrip van amoebiasis pathogenese verbeteren. Dit artikel presenteert een eenvoudig model dat de beoordeling van de eerste gastheer-pathogeen interacties en de parasiet invasie potentieel toestaat. Te analyseren parameters omvatten transepithelial elektrische weerstand interactie van EhCPADH112 epitheliale oppervlak receptoren, veranderingen in de expressie en localisatie van epitheliale junctie merkers en lokalisatie parasiet moleculen in epitheelcellen.

Voor een succesvolle E. histolytica cultuur, moeten twee belangrijke voorwaarden worden voldaan: de groei in axenische voorwaarden en oogst in logaritmische fase. Voorheen, culturen van E. histolytica werden gemakkelijk opgericht in samenwerking met bepaalde soorten bacteriën of trypanosomatids 22. Echter, tegenwoordig is het gebruikelijk om axenic teelt van deze parasiet betekent onbepaalde subkweek van amoeben in een omgeving zonder metaboliseren bacteriën, schimmels, protozoa, of metazo...

Watch this Scientific Journal Video about Analysis of the Epithelial Damage Produced by Entamoeba histolytica Infection at JoVE.com

Analysis of the Epithelial Damage Produced by Entamoeba histolytica Infection

Menselijke infectie door Entamoeba histolytica leidt tot amoebiasis, een belangrijke oorzaak van diarree in tropische landen. Infectie wordt gestart door pathogeen interactie met intestinale epitheelcellen, veroorzaken de opening van cel-cel contacten en bijgevolg diarree, soms gevolgd door leverbesmetting. Dit artikel geeft een model om de vroege gastheer-pathogeen interacties beoordelen in ons begrip van amoebiasis pathogenese verbeteren.

Analyse van de epitheelbeschadiging Geproduceerd door<em&gt; Entamoeba histolytica</em&gt; Infectie

Entamoeba histolytica is the causative agent of human amoebiasis, a major cause of diarrhea and hepatic abscess in tropical countries. Infection is ...

Research

JoVE Journal

Immunology and Infection

Analysis of the Epithelial Damage Produced by Entamoeba histolytica Infection

In vitro tRNA Methylation Assay with the Entamoeba histolytica DNA and tRNA Methyltransferase Dnmt2 (Ehmeth) Enzyme

In vitro tRNA Methylation Assay with the Entamoeba histolytica DNA and tRNA Methyltransferase Dnmt2 Ehmeth Enzyme

In vitro tRNA methylatie test met de Entamoeba histolytica DNA en tRNA methyltransferase Dnmt2 (Ehmeth) Enzym

Protozoan parasites are among the most devastating infectious agents of humans responsible for a variety of diseases including amebiasis, which is one of ...

Development of a Negative Selectable Marker for Entamoeba histolytica

Development of a Negative Selectable Marker for Entamoeba histolytica

Ontwikkeling van een negatieve selecteerbare marker voor Entamoeba histolytica</em

Entamoeba histolytica is the causative agent of amebiasis and infects up to 10% of the world's population. The molecular techniques that have enabled the ...

Application of a Mouse Ligated Peyer&#x2019;s Patch Intestinal Loop Assay to Evaluate Bacterial Uptake by M cells

Application of a Mouse Ligated PeyerÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s Patch Intestinal Loop Assay to Evaluate Bacterial Uptake by M cells

Toepassing van een muis geligateerd Peyer&#39;s Patch Intestinale Loop Assay om Bacteriële opname evalueren door M-cellen

The inside of our gut is inhabited with enormous number of commensal bacteria. The mucosal surface of the gastrointestinal tract is continuously exposed ...

Following Cell-fate in E. coli After Infection by Phage Lambda

Following Cell-fate in E. coli After Infection by Phage Lambda

Na Cell-lot in<em&gt; E. coli</em&gt; Na Infectie door Phage Lambda

The system comprising bacteriophage (phage) lambda and the bacterium E. coli has long served as a paradigm for cell-fate determination1,2. Following the ...

Flow Cytometric Isolation of Primary Murine Type II Alveolar Epithelial Cells for Functional and Molecular Studies

Flowcytometrische Isolatie van primaire Murine Type II alveolaire epitheelcellen voor Functionele en moleculaire studies

Throughout  the last years, the contribution of alveolar type II epithelial cells (AECII)  to various aspects of immune regulation in the lung has been ...

Analysis of the Solvent Accessibility of Cysteine Residues on Maize rayado fino virus Virus-like Particles Produced in Nicotiana benthamiana Plants and Cross-linking of Peptides to VLPs

Analysis of the Solvent Accessibility of Cysteine Residues on Maize rayado fino virus Virus-like Particles Produced in Nicotiana benthamiana Plants and Cross-linking of Peptides to VLPs

Analyse van de Solvent Toegankelijkheid van cysteïneresten op<em&gt; Maïs rayado fino virus</em&gt; Virus-achtige deeltjes, geproduceerd in<em&gt; Nicotiana benthamiana</em&gt; Plants and Cross-linking van peptiden aan VLP

Mimicking  and exploiting virus properties and physicochemical and physical  characteristics holds promise to provide solutions to some of  the world's ...

Capsular Serotyping of Streptococcus pneumoniae by Latex Agglutination

Capsular Serotyping of Streptococcus pneumoniae by Latex Agglutination

Capsulaire Serotypering van<em&gt; Streptococcus pneumoniae</em&gt; Door Latex Agglutinatie

Latex agglutination reagents are widely used in microbial diagnosis, identification and serotyping. Streptococcus pneumoniae (the pneumococcus) is a major ...

Rapid Analysis of Chromosome Aberrations in Mouse B Lymphocytes by PNA-FISH

Snelle analyse van chromosoomafwijkingen in Muis B-lymfocyten door PNA-FISH

Defective DNA repair leads to increased genomic instability, which is the root cause of mutations that lead to tumorigenesis. Analysis of the frequency ...

Use of the Soft-agar Overlay Technique to Screen for Bacterially Produced Inhibitory Compounds

Het gebruik van de Soft-agar Overlay techniek voor het screenen op bacterieel geproduceerd remmende verbindingen

The soft-agar overlay technique was originally developed over 70 years ago and has been widely used in several areas of microbiological research, ...

Imaging Ca2+ Responses During Shigella Infection of Epithelial Cells

Imaging Ca2+ Responses During Shigella Infection of Epithelial Cells

Imaging Ca2 + reacties tijdens Shigella besmetting van epitheliale cellen

Ca2+ is a ubiquitous ion involved in all known cellular processes. While global Ca2+ responses may affect cell fate, local variations in free Ca2+ ...