Conventional Laparoscopic vs Robotic Training: Which is Better for Naive Users? A Randomized Prospective Crossover Study.

OBJECTIVE: Robotic training (RT) using the da Vinci skills simulator and conventional training (CT) using a laparoscopic "training box" are both used to augment operative skills in minimally invasive surgery. The current study tests the hypothesis that skill acquisition is more rapid using RT than using CT among naive learners. DESIGN AND PARTICIPANTS: A total of 40 subjects without laparoscopic or robotic surgical experience were enrolled and randomized to begin with either RT or CT. Then, 2 specific RT tasks were reproduced for CT and repeated 5 times each with RT and CT. Time and quality indicators were measured quantitatively. A crossover technique was used to control for in-study experience bias. RESULTS: The tasks "pick and place jacks" (PP) and "thread the rings" (TR) were achieved faster with RT than with CT despite crossover (p < 0.0001). An RT-favoring difference was observed in speed for both tasks when changing modality. Percentage improvement with increasing trials was similar for RT and CT: RT completion time averaged 39 seconds and 211 seconds (PP and TR, respectively), compared with 65 seconds and 362 seconds when using CT (p < 0.0001); final improvement averaged 26% and 46% for RT (PP and TR, respectively) vs 31% and 47% for CT (p was 0.76 for PP and 0.20 for TR). Within the PP task, RT times averaged 41 seconds without previous CT experience vs 35 seconds with previous CT experience (p = 0.20); CT times averaged 61 seconds without and 69 seconds with previous RT experience (p = 0.48). Comparable times for the TR task were 212 seconds vs 216 seconds (p = 0.66) and 388 seconds vs 334 seconds (p = 0.17). Both instrument collisions and excessive force occurred more commonly for RT than for CT within the TR task (p < 0.0001). CONCLUSIONS: Speeds were faster overall with RT than with CT, but the percentage of speed improvement with trials was similar, suggesting similar learning curves, with minimal transfer effect appreciated.

Spontaneous tumor regression in a syngeneic rat model of liver cancer: implications for survival studies.

PURPOSE: To characterize tumor growth of N1S1 cells implanted into the liver of Sprague-Dawley rats to determine if this model could be used for survival studies. These results were compared with tumor growth after implantation with McA-RH7777 cells. MATERIALS AND METHODS: N1S1 or McA-RH7777 cells were implanted into the liver of Sprague-Dawley rats (n = 20 and n = 12, respectively) using ultrasound (US) guidance, and tumor growth was followed by using US. Serum profiles of 19 cytokines were compared in naive versus tumor-bearing rats. RESULTS: Both types of tumors were visible on US 1 week after tumor implantation, but the mean tumor volume of N1S1 tumors was larger compared to McA-RH7777 tumors (231 mm(3) vs 82.3 mm(3), respectively). Tumor volumes in both groups continued to increase, reaching means of 289 mm(3) and 160 mm(3) in N1S1 and McA-RH7777 groups, respectively, 2 weeks after tumor implantation. By week 3, tumor volumes had decreased considerably, and six tumors (50%) in the McA-RH7777 had spontaneously regressed, versus two (10%) in the N1S1 group. Tumor volumes continued to decrease over the following 3 weeks, and complete tumor regression of all tumors was seen 5 weeks and 6 weeks after tumor implantation in the McA-RH7777 and N1S1 groups, respectively. In an N1S1-implanted rat, multiple cytokines that have been shown to correlate with the ability of the tumor to survive in a hostile environment were increased by as much as 50%, whereas the average increase in cytokine levels was 90%. These findings suggest that the net cytokine environment favors an antitumor immune response. A similar trend was observed in a rat with a McA-RH7777 tumor, and the increase in cytokine levels was considerably more pronounced, with an average increase of 320%. CONCLUSIONS: The model of N1S1 cell implantation in the liver of Sprague-Dawley rats is not ideal for survival studies and should only be used with great caution in short-term studies that involve cancer therapies.

High expression of miR-210 predicts poor survival in patients with breast cancer: a meta-analysis.

BACKGROUND: Many microRNAs (miRNAs) exhibit altered expression levels in cancers, and they may be considered as valuable prognostic biomarkers for cancers. Here we aimed to summarize the recent advances in miR-210 involvement in human breast cancer and analyze the predicting role of miR-210 for survival. METHODS: A meta-analysis was performed by searching PubMed, Cochrane Library, and Science Direct databases. Data were extracted from studies comparing survival in patients with breast cancer having higher expression of miR-210 with those having lower expression. Pooled hazard ratios (HRs) and 95% confidence intervals (CI) were calculated. RESULTS: A total of 511 cases of breast cancer were involved for this global meta-analysis. For post-operational survival, the HR of higher miR-210 expression in breast cancer tissue was 3.39 (95% CI: 2.04-5.63, P<0.05), which could significantly predict poorer survival. CONCLUSIONS: High expression of miR-210 might predict poor survival in patients with breast cancer.

3-Bromopyruvate induces endoplasmic reticulum stress, overcomes autophagy and causes apoptosis in human HCC cell lines.

BACKGROUND: Autophagy, a cellular response to stress, plays a role in resistance to chemotherapy in cancer cells. Resistance renders systemic chemotherapy generally ineffective against human hepatocellular carcinoma (HCC). Recently, we reported that the pyruvate analog 3-bromopyruvate (3-BrPA) promoted tumor cell death by targeting GAPDH. In continuance, we investigated the intracellular response of two human HCC cell lines (Hep3B and SK-Hep1) that differ in their status of key apoptotic regulators, p53 and Fas. METHODS AND RESULTS: 3-BrPA treatment induced endoplasmic reticulum (ER) stress, translation inhibition and apoptosis based on Western blot and qPCR, pulse labeling, Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and active caspase-3 in both the cell lines. However, electron microscopy revealed that 3-BrPA treated SK-Hep1 cells underwent classical apoptotic cell death while Hep3B cells initially responded with the protective autophagy that failed to prevent eventual apoptosis. CONCLUSION: 3-BrPA treatment promotes apoptosis in human HCC cell lines, irrespective of the intracellular response.

The pyruvic acid analog 3-bromopyruvate interferes with the tetrazolium reagent MTS in the evaluation of cytotoxicity.

3-Bromopyruvate (3BrPA) is a pyruvate analog known for its alkylating property. Recently, several reports have documented the antiglycolytic and anticancer effects of 3BrPA and its potential for therapeutic applications. 3BrPA-mediated cytotoxicity has been evaluated in vitro by various methods including tetrazolium salt (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide)-based assays such as MTT, MTS, and so on. However, growing body of evidences has shown that tetrazolium reagent may interfere with the test compounds. In this study, we investigated whether the tetrazolium reagent interferes with the assessment of 3BrPA cytotoxicity. The results of the tetrazolium-based MTS assay were compared with 3 distinct cell viability detection methods, that is, Trypan Blue staining, ATP depletion, and Annexin V staining in 2 different cell lines, Vx-2 and HepG2. The MTS assay data showed false positive results by indicating increased cell viability at 1 mM and 2 mM 3BrPA whereas the other cell viability assays demonstrated that both Vx-2 and HepG2 cells are not viable at the same treatment conditions. In order to validate the direct interaction of 3BrPA with MTS reagent, we tested cell-free media incubated with different concentrations of 3BrPA. The results of cell-free media showed an increase in absorbance in a dose-dependent manner confirming the interaction of MTS with 3BrPA. Thus, our data clearly demonstrate that 3BrPA interferes with the accuracy of MTS-based cytotoxicity evaluation. Hence, we suggest that employing multiple methods of biochemical as well as morphological cytotoxicity assays is critical to evaluate 3BrPA-mediated cell death.

Percutaneous US-guided implantation of Vx-2 carcinoma into rabbit liver: a comparison with open surgical method.

PURPOSE: To evaluate technical feasibility and experimental usefulness of percutaneous US-guided implantation of Vx-2 carcinoma in rabbit liver. MATERIALS AND METHODS: Forty-eight New Zealand White male rabbits were used. Solid tumor mass of Vx-2 carcinoma was harvested from carrier rabbit, and minced tumor cells were implanted. Twenty-four rabbits underwent percutaneous US-guided tumor implantation, and the same number of rabbits underwent open laparotomy tumor implantation. Tested parameters included technical success, procedural time, amount of anesthesia, recovery time, complications, tumor size, and regional tumor seeding. RESULTS: A new percutaneous US-guided implantation was technically feasible in all rabbits. Evaluation parameters showed that the percutaneous US-guided implantation method is less invasive than the open laparotomy method. Targeting rate for left lateral lobe of implantation site was comparable in both methods (91.7% of percutaneous US-guided; 95.8% in open laparotomy). The success rate of tumor growth in the liver was 100% in both groups. However, in the group with US-guidance, tumor seeding developed more frequently in five of 24 rabbits (20.8%) than in open laparotomy group (2/24, 8.3%). Five rabbits had thoracoabdominal wall needle tract seeding, and two rabbits had tumor seeding at both thoracoabdominal wall and intraperitoneal space. CONCLUSIONS: Percutaneous US-guided implantation of Vx-2 carcinoma in rabbit liver is a less invasive alternative to open laparotomy, achieving equally successful tumor growth in the liver. Although percutaneous US-guidance implantation method may not be considered for long-term survival study design because of the possibility of tumor seeding, it can be considered for nonsurvival study design.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is pyruvylated during 3-bromopyruvate mediated cancer cell death.

BACKGROUND: The pyruvic acid analog 3-bromopyruvate (3BrPA) is an alkylating agent known to induce cancer cell death by blocking glycolysis. The anti-glycolytic effect of 3BrPA is considered to be the inactivation of glycolytic enzymes. Yet, there is a lack of experimental documentation on the direct interaction of 3BrPA with any of the suggested targets during its anticancer effect. METHODS AND RESULTS: In the current study, using radiolabeled ((14)C) 3BrPA in multiple cancer cell lines, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was identified as the primary intracellular target of 3BrPA, based on two-dimensional (2D) gel electrophoretic autoradiography, mass spectrometry and immunoprecipitation. Furthermore, in vitro enzyme kinetic studies established that 3BrPA has marked affinity to GAPDH. Finally, Annexin V staining and active caspase-3 immunoblotting demonstrated that apoptosis was induced by 3BrPA. CONCLUSION: GAPDH pyruvylation by 3BrPA affects its enzymatic function and is the primary intracellular target in 3BrPA mediated cancer cell death.

Development of a new orthotopic animal model of metastatic liver cancer in the rabbit VX2 model: effect on metastases after partial hepatectomy, intra-arterial treatment with 3-bromopyruvate and chemoembolization.

AIM: The aim of our study was to compare the influence of partial hepatectomy, intra-arterial treatment with 3-BrPA and TACE on regional and distant metastases. In order to achieve our objective, we tested the feasibility of both resection, intra-arterial therapy with 3-BrPA and TACE of VX2 liver cancer in New Zealand White rabbits. METHODS: VX2 tumors were implanted in the left lateral lobe of the liver of 20 rabbits. Tumors were allowed to grow for 14 days. Rabbits were divided in four groups. Group 1 (n=2) was sacrificed 14 days post implantation. Group 2, 3 and 4 (n=6) underwent left lateral hepatectomy, a 1 h intra-arterial infusion with 3-BrPA and TACE respectively. Animals in each group were further subdivided into three groups of two animals each corresponding to the time-point of sacrifice after the procedure (7, 14 and 21 days respectively). After sacrifice, organs were harvested, fixed and analyzed. RESULTS: Pathologic examination showed lung metastases in all 20 rabbits. Abdominal cavity dissemination was seen in five rabbits in Group 2, two rabbits in Group 3 and all rabbits in Group 4. Kidney metastases were seen in two rabbits treated with TACE. CONCLUSION: The VX2 rabbit model of liver cancer is a suitable model to compare the influence of partial hepatectomy, intra-arterial treatment with 3-BrPA and TACE on tumor recurrence in the form of regional and distant metastases. Our results indicate that intra-arterial delivery of 3-BrPA may result in a favorable metastatic profile when compared to both liver resection and TACE.

Targeting of VX2 rabbit liver tumor by selective delivery of 3-bromopyruvate: a biodistribution and survival study.

The aim of this study was to determine the biodistribution and tumor targeting ability of (14)C-labeled 3-bromopyruvate ([(14)C]3-BrPA) after i.a. and i.v. delivery in the VX2 rabbit model. In addition, we evaluated the effects of [(14)C]3-BrPA on tumor and healthy tissue glucose metabolism by determining (18)F-deoxyglucose (FDG) uptake. Last, we determined the survival benefit of i.a. administered 3-BrPA. In total, 60 rabbits with VX2 liver tumor received either 1.75 mM [(14)C]3-BrPA i.a., 1.75 mM [(14)C]3-BrPA i.v., 20 mM [(14)C]3-BrPA i.v., or 25 ml of phosphate-buffered saline (PBS). All rabbits (with the exception of the 20 mM i.v. group) received FDG 1 h before sacrifice. Next, we compared survival of animals treated with i.a. administered 1.75 mM [(14)C]3-BrPA in 25 ml of PBS (n = 22) with controls (n = 10). After i.a. infusion, tumor uptake of [(14)C]3-BrPA was 1.8 +/- 0.2% percentage of injected dose per gram of tissue (%ID/g), whereas other tissues showed minimal uptake. After i.v. infusion (1.75 mM), tumor uptake of [(14)C]3-BrPA was 0.03 +/- 0.01% ID/g. After i.a. administration of [(14)C]3-BrPA, tumor uptake of FDG was 26 times lower than in controls. After i.v. administration of [(14)C]3-BrPA, there was no significant difference in tumor FDG uptake. Survival analysis showed that rabbits treated with 1.75 mM 3-BrPA survived longer (55 days) than controls (18.6 days). Intra-arterially delivered 3-BrPA has a favorable biodistribution profile, combining a high tumor uptake resulting in blockage of FDG uptake with no effects on healthy tissue. The local control of the liver tumor by 3-BrPA resulted in a significant survival benefit.