Ectopic pregnancy in a case of congenital mullerian anomaly: a diagnostic dilemma

Ectopic or extrauterine pregnancy occurring in a case with mullerian defect is very rare and poses diagnostic challenges. Undescended and non-communicating fallopian tubes are extremely rare mullerian anomalies. Here authors present a case of ectopic pregnancy occurring in an undescended non-communicating fallopian tube in a patient with unicornuate uterus with absent horn, which was managed laparoscopically. A 32-year-old lady, diagnosed case of left unicornuate uterus with absent right horn, was referred to us with the suspicion of ruptured ectopic pregnancy. The abdominopelvic ultrasound showed a soft tissue lesion of size 32×24 mm, towards the right lateral pelvic wall near the iliac vessels, with increased vascularity on colour flow doppler.  The patient underwent laparoscopy which showed left sided unicornuate uterus with normal left tube and ovary. The right uterine horn was absent.  An undescended right ovary and tube were found attached to the peritoneum at the level of pelvic brim along the right lateral pelvic wall.  Right sided tubal ectopic pregnancy with rupture was present along with 300-350 cc of hemoperitoneum. The patient was treated with laparoscopic right sided total salpingectomy. In patients with unicornuate uterus and atypical presentation, ectopic pregnancy should be ruled out in an undescended non-communicating fallopian tube. Salpingectomy of incidentally diagnosed non-communicating fallopian tubes is recommended to prevent future ectopic pregnancy

Adventitial Delivery of Lentivirus-shRNA-ADAMTS-1 Reduces Venous Stenosis Formation in Arteriovenous Fistula

<div><p>Hemodialysis vascular access can develop venous neointimal hyperplasia (VNH) causing stenosis. Recent clinical and experimental data has demonstrated that there is increased expression of a disintegrin and metalloproteinase thrombospondin motifs-1 (ADAMTS-1) at site of VNH. The experiments outlined in the present paper were designed to test the hypothesis that targeting of the adventitia of the outflow vein of murine arteriovenous fistula (AVF) using a small hairpin RNA that inhibits ADAMTS-1 expression (LV-shRNA-ADAMTS-1) at the time of fistula creation will decrease VNH. At early time points, ADAMTS-1 expression was significantly decreased associated with a reduction in vascular endothelial growth factor-A (VEGF-A) and matrix metalloproteinase-9 (MMP-9) (LV-shRNA-ADAMTS-1 transduced vessels vs. controls). These changes in gene and protein expression resulted in favorable vascular remodeling with a significant increase in mean lumen vessel area, decrease in media/adventitia area, with a significant increase in TUNEL staining accompanied with a decrease in cellular proliferation accompanied with a reduction in CD68 staining. Collectively, these results demonstrate that ADAMTS-1 transduced vessels of the outflow vein of AVF have positive vascular remodeling.</p></div

Apoptosis is increased in the LV-shRNA-ADAMTS-1 transduced vessels.

<p><b>A</b>) is a representative section from TUNEL staining at the venous stenosis of the LV-shRNA-ADAMTS-1 (<b>LV</b>) and control shRNA(<b>C</b>) transduced control vessels at day 14. Nuclei staining brown are positive for TUNEL. Negative control is shown where the recombinant terminal deoxynucleotidyl transferase enzyme was omitted. All are 40X. Scale bar is 50-μM. * Indicates the lumen. Arrow indicates TUNEL positive cells. Pooled data for <b>LV</b> and <b>C</b> transduced vessels for TUNEL index at the outflow vein of the <b>LV</b> and <b>C</b> transduced vessels at day 7 (<b>D7</b>), 14 (<b>D14</b>), and 21 (<b>D21</b>) is shown in <b>(B)</b>. This demonstrates a significant increase in the mean TUNEL index at day 14 (P<0.01) and day 21 (P<0.05) in the <b>LV</b> group when compared to <b>C</b>. Each bar shows mean ± SEM of 3–6 animals per group. Two-way ANOVA followed by Student <i>t</i>-test with post hoc Bonferroni's correction was performed. Significant difference from control value was indicated by *<i>P</i><0.05 or **<i>P</i><0.01.</p

Cellular proliferation is decreased in LV-shRNA-ADAMTS-1 transduced vessels.

<p><b>A</b>) is representative sections after Ki-67 staining at the venous stenosis of the LV-shRNA-ADAMTS-1 (<b>LV</b>) and control shRNA (<b>C</b>) at day 7. Nuclei staining brown are positive for Ki-67. IgG antibody staining was performed to serve as negative control. All are 40X. Scale bar is 50-μM. * indicates the lumen. Pooled data for Ki-67 index at the outflow vein of the <b>LV</b> and <b>C</b> transduced vessels at day 7 (<b>D7</b>), 14 (<b>D14</b>), and 21 (<b>D21</b>) is shown in (<b>B</b>). By day 7, there is a significant decrease in the mean Ki-67 index in the <b>LV</b> transduced vessels when compared to <b>C</b> at day 7 (P<0.01). By day 14, the mean Ki-67 index in the <b>LV</b> transduced vessels is significantly increased when compared to <b>C</b> (P<0.01). Each bar shows mean ± SEM of 4-6 animals per group. Two-way ANOVA followed by Student <i>t</i>-test with post hoc Bonferroni's correction was performed. Significant difference from control value was indicated by *<i>P</i><0.01.</p

ADAMTS-1 expression is reduced in LV-shRNA-ADAMTS-1 transduced vessels.

<p><b>A</b>) is representative sections using confocal microscopy from the outflow vein after transduction with LV-shRNA-ADAMTS-1 (<b>LV</b>) and control shRNA (<b>C</b>) at day 7 (<b>D7</b>) and 14 (<b>D14</b>). Both the <b>C</b> and <b>LV</b> have a GFP tag. First column shows DAPI intensity indicated by blue staining nuclei. Second column shows GRP intensity. Magenta staining cells are GFP positive. Third column depicts merged images. These sections demonstrate that there is <b>LV</b> or <b>C</b> delivery to the adventitia and endothelium of the vessel wall. By day 14, the gfp signal decreases and is not detectible. <b>B</b>) is the pooled data of the mean gene expression of ADAMTS-1 at the outflow vein after transduction with LV-shRNA-ADAMTS-1 (<b>LV</b>) compared to control shRNA (<b>C</b>) using qPCR analysis at day 7 (<b>D7</b>), 14 (<b>D14</b>), and 21 (<b>D21</b>). This demonstrates a significant reduction in the mean ADAMTS-1 expression in the <b>LV</b> transduced vessels when compared to <b>C</b> vessels at <b>D7</b> (P<0.0001). <b>C</b>) is representative sections from ADAMTS-1 staining at the outflow vein of <b>LV</b> transduced vessels when compared to <b>C</b> vessels at day 14. Cells staining brown are positive for ADAMTS-1. IgG antibody staining was performed to serve as negative control. <b>D</b>) is the pooled data of the mean ADAMTS-1 staining at the outflow vein after transduction with <b>LV</b> compared to <b>C</b> at day 7 (<b>D7</b>), 14 (<b>D14</b>), and 21 (<b>D21</b>). This shows that by <b>D14</b>, there is a significant reduction in the mean ADAMTS-1 staining in the <b>LV</b> transduced vessels when compared to <b>C</b> (P<0.0001). All are 40X. Scale bar is 50-μM. * Indicates the lumen. Each bar shows mean ± SEM of 4-6 animals per group (<b>B, D</b>). Two-way ANOVA followed by Student <i>t</i>-test with post hoc Bonferroni's correction was performed. Significant difference from control value was indicated by ^##<i>P</i><0.0001.</p

Q-PCR primers used.

<p>Q-PCR primers used.</p

MMP-9 expression is decreased in LV-shRNA-ADAMTS-1 transduced vessels.

<p><b>A</b>) is pooled data for qPCR analysis for MMP-9 expression of the LV-shRNA-ADAMTS-1 (<b>LV</b>) and control shRNA (<b>C</b>) transduced vessels at day 7 (<b>D7</b>), 14 (<b>D14</b>), and 21 (<b>D21</b>). This demonstrates a significant reduction in the average expression of MMP-9 in the <b>LV</b> transduced vessels when compared to <b>C</b> at day 7 (P<0.05) and 14 (P<0.05) with a significant increase in MMP-2 by day 28 (P<0.001). <b>B</b>) are representative sections from MMP-9 staining at the venous stenosis of the <b>LV</b> and <b>C</b> transduced vessels at day at day 14. Cells staining brown are positive for MMP-9. IgG antibody staining was performed to serve as negative control. <b>C</b>) is pooled data for the average MMP-9 expression at the outflow vein of the <b>LV</b> and <b>C</b> transduced vessels at day 7 (<b>D7</b>), 14 (<b>D14</b>), and 21 (<b>D21</b>). This shows that there is a significant reduction in the mean MMP-9 staining in the <b>LV</b> transduced vessels when compared to controls by day 7 (P<0.01) and day 14 (P<0.05). All are 40X. Scale bar is 50-μM. * Indicates the lumen. Each bar shows mean ± SEM of 3-6 animals per group (<b>A, C</b>). Two-way ANOVA followed by Student <i>t</i>-test with post hoc Bonferroni's correction was performed. Significant difference from control value was indicated by *<i>P</i><0.05 or ^#<i>P</i><0.001.</p