Temporins are effective in vivo when given 7 days after experimental infection.

<p>Mice were infected with a sub-lethal dose (10⁵ CFU/mouse) of <i>S.aureus</i> A170 (A) or a sub-lethal dose(10⁵ CFU/mouse) of <i>S.enterica</i> serovar Paratyphi B (B–C) and 6 days later were treated with the 35/75 pool of temporins. A: kidneys from mice infected with <i>S.aureus</i> A170 (closed box) and kidneys from mice infected with <i>S.aureus</i> A170 and treated after 6 days with TA plus TB-YK (open rhomb); B: gastro instestinal tract from mice infected with <i>S.enterica</i> serovar Paratyphi B (closed box ) and gastro intestinal tract from mice infected with <i>S.enterica</i> serovar Paratyphi B and treated after 6 days with TA plus TB-YK (open rhomb); C: liver from mice infected with <i>S.enterica</i> serovar Paratyphi B (10⁵ CFU/mouse) (closed box) and liver from mice infected with <i>S.enterica</i> serovar Paratyphi B (10⁵ CFU/mouse) and treated after 6 days with the TA plus TB-YK (open rhomb).</p

Synergism between TA and TB-YK temporins.

<p>The diameter of the inibition was maesured after 24 h incubation.</p

Temporins initiate their antibacterial activity by drillings holes in the bacterial membrane.

<p>A: <i>S.aureus</i> A170, untreated; B: <i>S aureus</i> A 170 treated for 10 min with TA (8 µg/ml) plus TB-YK (5 µg/ml) (B); C: <i>S.enterica</i> serovar Paratyphi B untreated; D: <i>S.enterica</i> serovar Paratyphi B treated for 10 min with TA (100 µg/ml) plusTB-YK (4 µg/ml).</p

Mice infected with GFP-labelled bacteria were analysed using the Leica macrofluo instrument (Wetzlar, Germany) equipped with the Leica application suite 3.1.0 software.

<p>A: liver from mice infected with <i>S. enterica</i> serovar Paratyphi B; B: liver from mice infected with <i>S. enterica</i> serovar Paratyphi B and immediately treated with TA plus TB-YK; C: gastro instestinal tract from mice infected with <i>S. enterica</i> serovar Paratyphi B; D: gastro instestinal tract from mice infected with <i>S. enterica</i> serovar Paratyphi B and immediately treated with TA plus TB.</p

Expression levels of cytokine genes in mice infected with <i>S.enterica</i> serovar Paratyphi B (10⁵ CFU/mouse) and 6 days lose treated with temporins (35/75 pool).

<p>Each hystogram represents the mean value measured in 2 mice, each one tested in triplicate.</p

CD spectra of: TB (blue, continuous line), TB-YKβA (red continuous line) in Sodium Phosphate buffer pH 7.4 (-); TB (blue dashed line) and TB-YKβA (red dashed line) in SodiumPhosphate buffer+SDS pH 7.4.

<p>CD spectra of: TB (blue, continuous line), TB-YKβA (red continuous line) in Sodium Phosphate buffer pH 7.4 (-); TB (blue dashed line) and TB-YKβA (red dashed line) in SodiumPhosphate buffer+SDS pH 7.4.</p

Expression levels of cytokine genes in mice infected with <i>S.enterica</i> serovar Paratyphi B (10⁶ CFU/mouse) (In) or infected with <i>S.enterica</i> serovar Paratyphi B (10⁶ CFU/mouse) and immediately after treated with the 35/75 pool of temporins (Tr).

<p>Expression levels of cytokine genes in mice infected with <i>S.enterica</i> serovar Paratyphi B (10⁶ CFU/mouse) (In) or infected with <i>S.enterica</i> serovar Paratyphi B (10⁶ CFU/mouse) and immediately after treated with the 35/75 pool of temporins (Tr).</p

Detection of total, viable and membrane-perturbed bacteria using temporin A or temporin B labeled with rhodamine isothiocyanate or fluorescein isothiocyanate respectively.

<p>A- B-C: <i>S.aureus</i> A 170 plus TA-Rho 6 µg plus TB-Fluo 9 µg; D-E-F: <i>S.aureus</i> A170 plus fluorescein isothiocyanate o rhodamine isothiocyanate (control); G-H-I: <i>S.enterica</i> serovar Paratyphi B plus TA-Rho 15 µg plus TB-Fluo 20 µg; L-M-N: <i>S.enterica</i> serovar Paratyphi B plus fluorescein isothiocyanate o rhodamine isothiocyanate (control).</p

In vivo antibacterial activity of temporins.

<p>The 35/75 pool of temporins, given concurrently with bacteria, rescue 100% of mice infected with a lethal dose of <i>S. aureus</i> A170 10⁷ CFU/mouse (A: mice infected with <i>S.aureus</i> A170 and immediately treated with TA plusTB-YK (closed box) and mice infected with <i>S.aureus</i> A170 (open triangle) or a lethal dose of <i>S. enterica</i> serovar Paratyphi 10⁶ CFU/mouse (B: Mice infected with <i>S.enterica</i> serovar Paratyphi B and immediately treated with TA plus TB-YK (closed box) and mice infected with <i>S.enterica</i> serovar Paratyphi B (open triangle)</p

Temporins sterilize mice sperimentally infected with <i>S aureus</i> A170 or <i>S.enterica</i> serovar Paratyphi B.

<p>The 35/75 pool of temporins, given concurrently with bacteria, sterilize mice infected with a lethal dose (10⁷ CFU/mouse) of <i>S.aureus</i> A170 (A: kidneys from mice infected with <i>S. aureus</i> A170 (closed rhomb) and kidneys from mice infected with <i>S.aureus</i> A170 and immediately after treated with TA plus TB-YK (open box), or a lethal dose (10⁶ CFU/mouse) of <i>S.enterica</i> serovar Paratyphi B (B: gastro instestinal tract from mice infected with <i>S.enterica</i> serovar Paratyphi B (closed rhomb) and gastro intestinal tract from mice infected with <i>S.enterica</i> serovar Paratyphi B and immediately treated with TA plus TB-YK (open box); (C: liver from mice infected with <i>S.enterica</i> serovar Paratyphi B (closed rhomb) and liver from mice infected with <i>S.enterica</i> serovar Paratyphi B and immediately treated with TA plus TB-YK (open box).</p