Preeclamptic features in CBA/J x DBA/2 matings.

<p>Increased fetal resorption frequency (A) and decreased litter size (B)) were observed only in first pregnancies in DBA/2 mated CBA/J mice. Normal pregnancies (not different from control matings CBA/J x BALB/c) were observed in second (2^nd P) and third (3^rd P) pregnancies with the same male. Data are mean values plus or minus SD. (C) In CBA/J x DBA/2 mice, a time-course increase in urine albumin to creatinine ratio (ACR), that reaches statistic significance at day 10 of pregnancy, was observed (*p<0.05 vs CBA/J x DBA/2). Albuminuria was not observed in control matings CBA/J x BALB/c mice. N = 6-8 mice/experimental group (Di) Transmission electron micrographs of glomeruli from DBA/J mated CBA/J mice (original magnification 10,000x) show swelling of glomerular endothelial cells and loss of fenestrations. Numerous red blood cells trapped between swollen endothelial cells were observed occluding the glomerular capillaries. No signs of endothelial injury, well preserved fenestrations and widely patent glomerular capillary lumina are observed in kidneys from CBA/J x BALB/c mice (control group) (Ei). (Dii) Kidney perfusion studies reveal diminished blood perfusion (the fluorescent tracer did not accumulate in the glomerular capillaries) in glomeruli from CBA/J x DBA/2 mice with abnormal pregnancies compared to control CBA/J x BALB/c matings (Eii). (Diii) Increased fibrin staining in glomeruli from CBA/J x DBA/2 mice when compared to CBA/J x BALB/c mice (Eiii). (Div and Eiv) Jones methenamine silver staining. Diffusely and irregularly thickened glomerular basement membranes were observed in some glomeruli in CBA/J x DBA/2 mice (20%) (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013663#pone-0013663-g001" target="_blank">Fig 1D</a>iv) compared to BALB/c mated CBA/2 females (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013663#pone-0013663-g001" target="_blank">Fig 1E</a>iv). Kidneys from 5-6 mice were studies in each experimental group.</p

MAP and contractile response to angiotensin II in CBA/J x DBA/2 mice.

<p>(A) No significant increase in MAP was observed in CBA/J x DBA/2 compared to CBA/J x BALB/c control matings. Data were expressed as change in MAP from mating day. CBA/J x DBA/2 mice showed an increase response to an acute a bolus injection of AngII compared to CBA/J x BALB/c mice (*p<0.01). The response to Ang II was expressed as the mean change in MAP from basal values before Ang II injection. (B) Aortic rings from CBA/J x DBA/2 and CBA/J x BALB/c contracted in response to Ang II (* p<0.05). Aortic rings from CBA/J x DBA/2 mice showed increased reduction of the inner diameter in response to Ang II (# different from CBA/J x BALB/c. P<0.05). N = 8–10 mice/experimental group. (C) Staring at day 11 of pregnancy, plasma leptin levels were increased in CBA/J x DBA/2 mice when compared to control CBA/J x BALB/c matings (* p<0.01). This increase was maintained until delivery. Leptin levels in CBA/J x DBA/2 and control matings drop to pre-pregnancy values after parturition. N = 6–8 mice/experimental group.</p

Effect of ²²⁵Ac-E4G10 therapy on tumor histology, vascularity and apoptosis.

<p>A, Light microscopy depicting numerous RBC-filled vascular spaces (arrows) in dual control tumor and fewer, but relatively normal-looking vessels (arrowheads) in the ²²⁵Ac-E4G10 treated tumor. B, Top: Immunohistochemical staining of tumor-sections for vWF, an endothelial cell marker (top). TUNEL staining of tumor sections to detect apoptosis (bottom). Quantification of vWF staining (C) and apoptosis (D) in 4 randomly selected fields. Data are mean ± S.E.M.</p

²²⁵Ac-E4G10 treatment results in a relatively normal remaining tumor vasculature.

<p>A, Greater coverage of tumor blood vessels (CD31 positive) by pericytes (α-SMA-positive cells) in ²²⁵Ac-E4G10 treated tumor relative to dual control. B, Transmission electron micrographs of blood vessels in dual control and ²²⁵Ac-E4G10 treated tumor. The dual control tumor contains extravasated RBC-filled vascular spaces that are not lined with endothelial cells, whereas blood vessels in ²²⁵Ac-E4G10 treated tumor display a continuous endothelial lining (arrow) resting on a basement membrane (BM) that is shared with the surrounding pericyte. Scale bar, 50 µm</p

²²⁵Ac-E4G10 therapy inhibits the growth of LnCap prostate tumors.

<p>A, Flow cytometric analysis depicting the lack of E4G10 binding to LnCap cells; J591, mouse-anti prostate specific membrane antigen is the positive control. Mouse and rat isotype controls were also evaluated. B, Photographs of in situ (left) and excised tumor (right) in a representative dual control and ²²⁵Ac-E4G10 treated animal. C, Tumor volume in various treatment groups at described time-points. D, Serum prostate specific antigen (PSA) levels in the three treatment groups at 22 days post-implantation with 5 million LnCap cells. E, Kaplan Meier curve showing enhancement of survival with ²²⁵Ac-E4G10 treatment. Data in C, D are mean ± S.E.M. <i>Scale bar</i>, 1 cm.</p

A combination of ²²⁵Ac-E4G10 with paclitaxel enhances the anti-tumor response.

<p>A, Tumor volume in the four treatment groups over time. Data are mean ± S.E.M. B, Kaplan Meier survival curve of treated animals showing significant enhancement of animal survival when ²²⁵Ac-E4G10 therapy is followed by a course of paclitaxel. C, Absence of histopathologic damage in normal organs, assessed 10 days after cessation of ²²⁵Ac-E4G10 treatment.</p