Journal: International Journal of Peptides

Article Title: Activity of Cathelicidin Peptides against Simkania negevensis

doi: 10.1155/2011/708710

Figure Lengend Snippet: Activity of cathelicidin peptides at different concentrations on the infectivity of Simkania negevensis.

Article Snippet: The reference strain Z of S. negevensis (American Type Culture Collection VR-1471) was grown in LLC-MK2 cells [ ] in 6-large well plates and elementary bodies (EBs) were purified by use of sucrose gradients [ ], resuspended in sucrose-phosphate-glutamic acid buffer (SPG) pH 7.4 (75 g sucrose, 87 mL 0.2 M Na 2 HPO 4 , 13 mL 0.2 M NaH 2 PO 4 , 0.72 g L-glutamic acid), and frozen in aliquots at −80°C.

Techniques: Activity Assay, Infection

Journal: PLoS ONE

Article Title: Lack of Effective Anti-Apoptotic Activities Restricts Growth of Parachlamydiaceae in Insect Cells

doi: 10.1371/journal.pone.0029565

Figure Lengend Snippet: S2 cells were either left untreated or were treated with SPG buffer, amoebal lysate, infectious Parachlamydiaceae ( Pa. acanthamoebae or P. amoebophila ; MOI 5) in the absence (Inf) or presence of the protein synthesis inhibitor doxycycline (Inf-Dox), heat-inactivated bacteria (Hi), UV- inactivated bacteria (UVi), a sterile-filtrate of the suspension of purified infectious bacteria (Filtrate) or a supernatant collected 48 h p.i. from an infected (MOI 5) apoptotic culture (Supernatant). For comparison, cells treated with infectious S. negevensis Z (MOI 5 or 50, as indicated) are shown. After 48 h incubation, DNA was stained with DAPI and the proportion of nuclei with altered morphology was determined. Mean values and standard deviations of six replicates (derived from three independent experiments) are shown. At least 500 nuclei per replicate were considered. Statistically significant differences compared to the untreated cells are indicated (ANOVA & Scheffé; ***, p≤0.001; **, p≤0.01; *, p≤0.05).

Article Snippet: The following fluorescently labelled (Cy3, Cy5, or Fluos) probes (Thermo Fisher Scientific, Germany) were applied in this study: EUK-516 ( 5′- ACCAGACTTGCCCTCC -3′ ) for the detection of eukaryotic host cells, Chls0523 ( 5′-CCTCCGTATTACCGCAGC-3′ ) in combination with the competitor probe ( 5′-CCTCCGTATTACCGCGGC-3′ ) as general probe for the detection of all chlamydiae used in this study, and UV7-763 ( 5′-TGCTCCCCCTTGCTTTCG-3′ ) and Simneg183 ( 5′-CAGGCTACCCCAGCTC-3′ ) for the specific detection of Pa. acanthamoebae UV7 and S. negevensis Z, respectively.

Techniques: Purification, Infection, Incubation, Staining, Derivative Assay