Purpose: To evaluate agonistic TRA-8 monoclonal antibody to human death receptor 5 (DR5) and gemcitabine in vitro and in an orthotopic pancreatic cancer model. Experimental Design: Pancreatic cancer cell lines were screened for DR5 expression, cytotoxicity, and apoptosis induced by TRA-8, gemcitabine, or gemcitabine and TRA-8. An orthotopic model of pancreatic cancer was established in severe combined immunodeficient mice. Mice were treated with TRA-8, gemcitabine, or a combination for one or two cycles of therapy. Tumor growth (ultrasound) and survival were analyzed. Results: All five pancreatic cancer cell lines showed DR5 protein expression and varying sensitivity to TRA-8 -mediated cytotoxicity. MIA PaCa-2 cells were very sensitive to TRA-8, moderately resistant to gemcitabine, with additive cytotoxicity to the combination. S2-VP10 cells were resistant to TRA-8 and sensitive to gemcitabine with synergistic sensitivity to the combination. Combination treatment in vitro produced enhanced caspase-3 and caspase-8 activation. A single cycle of therapy produced comparable efficacy for single-agent TRA-8 and the combination of TRA-8 and gemcitabine, with significant reduction in tumor size and prolonged survival compared with gemcitabine alone or control animals. With two cycles of therapy, TRA-8 and combination therapy produced enhanced inhibition of tumor growth compared with single-agent gemcitabine or untreated animals. However, the combination regimen showed enhanced survival as compared with single-agent TRA-8. Conclusions: Pancreatic cancer cell lines express varying levels of DR5 and differ in their sensitivity to TRA-8 and gemcitabine-induced cytotoxicity. TRA-8 with two cycles of gemcitabine therapy produced the best overall survival.

Purpose: Evaluate the response of human pancreatic cancer cell lines and orthotopic tumors to TRA-8, an agonistic antibody to death receptor 5, in combination with irinotecan (CPT-11). Experimental Design: MIA PaCa-2 and S2VP10 cells were treated withTRA-8 and/or CPT 11. Cell viability was determined by ATP assay. JC-1 mitochondrial depolarization and Annexin V assays confirmed cell death by apoptosis. Immunoblotting was used to evaluate protein changes. MIA PaCa-2 cells were injected into the pancreas of severe combined immunodeficient mice. Mice underwent abdominal ultrasound to quantitate tumor size before and after treatment with twice weekly injections of 200 Ag TRA-8 and/or 25 mg/kg CPT-11 for one or two treatment cycles, each lasting 2 weeks. Results: MIA PaCa-2 cells were more sensitive to TRA-8 and showed additive cytotoxicity, whereas S2VP10 cells showed synergistic cytotoxicity when treated with TRA-8 and CPT-11. Cell death occurred via apoptosis with increased cleavage of caspase-3, caspase-8, and caspase-9 and proapoptotic proteins Bid and poly(ADP)ribose polymerase after combination treatment compared with either agent alone. XIAP and Bcl-XL inhibitors of apoptosis were down-regulated. After a single cycle of in vivo combination therapy, tumor sizes had diminished significantly (P < 0.001) at 8 days posttreatment compared with no treatment, CPT-11, and TRA-8; and there was a 50-day increase in survival with combination treatment over untreated controls (P = 0.0002), 30 days over TRA-8, and a 36-day increase over CPT-11 monotherapy (P = 0.0003). With two cycles of TRA-8/CPT-11 treatment, mean survival time increased significantly (P < 0.001) to 169 days versus untreated controls, TRA-8 or 121, or 108 days, respectively). Conclusions: Combination TRA-8 and CPT-11 therapy produced enhanced cytotoxicity and survival in the MIA PaCa-2 orthotopic model of pancreatic cancer.

Superficial inferior epigastric artery (SIEA) flap breast reconstruction has advantages over deep inferior epigastric perforator flap (DIEP) and muscle sparing transverse rectus abdominus myocutaneous flap (TRAM) reconstructions with less donor site morbidity and less complicated flap dissection. Not all patients have an adequate SIEA and superficial inferior epigastric vein (SIEV) to support free tissue breast reconstruction, and dissection of the SIEA in all patients can be time consuming. Preoperative computed tomography (CT) angiograms can be used to identify the SIEA and SIEV in patients planning to undergo free abdominal tissue breast reconstruction and direct more efficient dissection in patients with a large SIEA. Retrospective analysis of free abdominal tissue flap breast reconstruction from a single plastic surgeon was performed. All patients undergoing free abdominal tissue breast reconstruction had a preoperative CT angiogram using a protocol for the evaluation of the abdominal wall perforating arteries. CT scans were reviewed by the surgeon preoperatively and evaluated for the presence, caliber, and image quality of the SIEA and SIEV. All patients, regardless of CT angiogram findings, had operative dissection and evaluation of the SIEA and SIEV. A total of 177 free flaps were performed on 113 patients who underwent preoperative CT angiogram and free abdominal tissue breast reconstruction. Of them, 64 patients had bilateral breast reconstruction. Twelve SIEA flaps (10.6%) were performed on 12 patients. During preoperative CT angiographic evaluation, 49 patients (43%) were noted to have at least one visible SIEA, whereas only 24 of those patients (21%) were felt to have an SIEA of adequate caliber. No flaps were lost during the postoperative period. All 12 SIEA flaps performed had an adequate SIEA when observed on preoperative CT angiogram. Overall, 50% of patients found to have at least one adequate SIEA on CT angiogram had a single breast reconstructed with an SIEA flap. If the SIEA was not visualized on CT angiogram, no usable SIEA was found during surgery. Preoperative CT angiogram of the abdominal wall perforating arteries can help predict which patients may have adequate anatomy for an SIEA-based free flap. This information may help direct more efficient dissection of the abdominal flaps by selecting out patients who do not have an adequate SIEA.

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Background:The ability to salvage the mangled lower extremity is both technically challenging and time consuming. It requires the collaborative efforts among multiple surgical specialties in addition to comprehensive post-traumatic wound follow-up. Our institution has integrated a dynamic effort among these specialists in the planning and facilitating a successful limb salvage program with creation of a mangled extremity algorithm. An integral part in this process is the vascular inflow to prepare coverage for large tissue defects lacking adequate recipient targets. Utilization of long saphenous arteriovenous (AV) loop has been cited with minimal data available using larger inflow vessels in the acute trauma setting. We performed a retrospective review and describe our early experience using our protocol with AV loop creation with free flap reconstruction to salvage traumatic leg injuries. Using the data, we sought to develop a mangled extremity protocol for trauma centers to guide mangled limb salvage. Methods: Since June 2016, 398 patients were admitted to our level II trauma facility with isolated traumatic wounds to the lower extremities. Thirty-one limbs were deemed mangled in which 21 received primary amputations due to multiple factors. Ten patients admitted from the trauma service with isolated mangled lower extremities injuries were identified for review. All 10 patients sustained severe crush injuries with large soft tissue defects and decreased perfusion for healing but deemed salvageable by multispecialty assessment. Mangled extremity severity scores were tabulated. Patients age ranged from 21e44 years, with 8 men and 2 women. Repeated debridements until successful sterilization of the wounds were accomplished. Ten long saphenous vein AV loops were anastomosed to the at or above knee popliteal vessels for free flap reconstruction. All patients were followed post-AV loop creation for vascular complications and wound assessments. Results: All 10 patients had sterilization of the wounds with repair of the fracture site before vascular reconstruction. Mean debridement to surgical site sterilization was 4.3 washouts (range 2e7). Successful AV loop creation with long saphenous vein was completed in 100% of patients without vascular complications nor steal events. Free flap tissue transfers directly connected to the loop were completed using 6 rectus abdominis, 3 latissimus dorsi, and 1 anterior thigh graft within 10 days of its creation. Patency rates of the AV loop was 100% with 10 successful flap transfers and 90% amputation free survival. One flap did not survive due to recurrent bacterial infection of the hardware. The 9 patients with successful procedures reached preoperative ambulatory status within 3 months after their final surgery. At 24 months follow-up, 90% amputation free survival is still maintained.

Objective. The aim of this study was to determine if early initiation of oral/enteral nutrition in burn patients minimizes the drop in fT3 levels, reduces the potential for euthyroid sick syndrome (ESS), and shortens the length of hospital stay (LHS). Subjects and Methods. We retrospectively evaluated the statistical association of serum fT3, fT4, and TSH at the first (2nd–5th day) and second sample collection (9th–12th day) after the burn injury in 152 burn patients. Three groups were established depending on time of initiation of the oral/enteral nutrition: <24 h before the injury (Group 1), 24–48 h after the injury (Group 2), and >48 h after the injury (Group 3). Results. They were expressed as mean ± standard deviation. We found that LHS and the fT3 levels were statistically different in the 3 groups. The LHS (in days) was, respectively, in each group, 16.77 ± 4.56, 21.98 ± 4.86, and 26.06 ± 5.47. Despite the quantifiable drop in fT3, ESS was present only at the first sample collection (2.61 ± 0.92 days) in Group 3, but there was no group with ESS at the second sample collection (9.89 ± 1.01 days). Our data suggest that early initiation of nutritional supplementation decreases the length of hospitalization and is associated with decreasing fT3 serum concentration depression. Conclusion. Early initiation of oral/enteral nutrition counteracts ESS and improves the LHS in burn patients.

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Supplementary Table 1, Supplementary Figures 1-3 from TRA-8 anti-DR5 monoclonal antibody and gemcitabine induce apoptosis and inhibit radiologically validated orthotopic pancreatic tumor growth