Journal of Surgical Research - Articles in Press

Transplantation of Cbfa1-Overexpressing Adipose Stem Cells Together With Vascularized Periosteal Flaps Repair Segmental Bone Defects - Uncorrected Proof

Jianjun Li, Qun Zhao, Enbo Wang, Chuanhui Zhang, Guangbin Wang, Quan Yuan — 2012-01-04

Background: Segmental bone defect is still a challenge to orthopedic surgeons. Currently available therapies for segmental bone defects have some drawbacks. Tissue engineering using pluripotent stem cells is a new, promising method for bone repair. The present study aims to promote the effect of bone defect repair using the tissue engineered bone in combination with vascularized periosteal flaps.Methods: The adenoviral vector carrying Cbfa1 transduced rabbit adipose-derived mesenchymal stem cells and gene modified tissue engineering bone (GMB) were constructed. Rabbits with radial defects were implanted with the GMB together with vascularized periosteum (group A); or GMB with free periosteum (group B); or GMB (group C), and scaffold (group D). The bone repair effect was evaluated at 4, 8, or 12 wk, respectively, after the operations.Results: Cbfa1 proteins were strongly expressed in adipose stem cells (ADSCs) that formed a stratified network on the inner surface of the polylactic acid/ polycaprolacton (PLA/PCL) pores. Bone repair was well achieved in the rabbits treated with the Cbfa1-expressing ADSCs and vascularized flap that was markedly better than those treated with either Cbfa1-expressing ADSCs alone or with vascularized flap alone.Conclusions: Combination with implanting the Cbfa1 gene-modified tissue-engineered bone and vascularized periosteum can better repair the segmental bone defects by stimulating osteogenesis, osteoinduction, and osteoconduction than using either one of the approaches.

Berberine Suppresses the TPA-Induced MMP-1 and MMP-9 Expressions Through the Inhibition of PKC-α in Breast Cancer Cells - Uncorrected Proof

2012-01-03

Background: Berberine (BBR) is one of the major alkaloids, and it has been reported to have a variety of pharmacologic effects, including inhibition of cell cycle progression. Here, we investigated the effect of BBR on the MMP-1 and MMP-9 expressions, which are predictors of metastasis and invasion in breast cancer cells.Methods: MMP-1 and MMP-9 mRNA expressions were analyzed by real-time PCR. The levels of MMP-1 protein and PKC-α phosphorylation were detected by Western blotting. MMP-9 protein expression was detected by gelatin zymography. Cell cycle was analyzed by FACS analysis. PKC-α knock-down was examined by PKC-α siRNA transfection.Results: The basal levels of both the MMP-1 and MMP-9 mRNA expressions were decreased by BBR treatment in a dose-dependent manner. In contrast, TPA, which is a tumor promoter, significantly increased the levels of the MMP-1 and MMP-9 mRNA and protein expressions in the MCF-7 breast cancer cells. We also observed that the TPA-induced MMP-1 and MMP-9 mRNA and protein expressions were prevented by BBR treatment. In addition, the TPA-induced MMP-1 and MMP-9 expressions were completely decreased by Go6983 and PKC-α siRNA, respectively. TPA-induced PKC-α phosphorylation was dose-dependently decreased by BBR treatment.Conclusion: The TPA-induced PKC-α phosphorylation is suppressed and then the MMP-1 and MMP-9 expressions are also inhibited by berberine. Therefore, we suggest that berberine may be used as a candidate drug for the inhibition of metastasis of human breast cancer.

Overexpression of Programmed Death Ligand 1 in Dendritic Cells Inhibits Allogeneic Lymphocyte Activation in Mice - Uncorrected Proof

Wenzhi Li, Xiang Wang, Renfu Chen, Haitao Zhu, Gang Chen, Xiaoqing Sun — 2012-01-03

Background: Co-stimulatory molecules are pivotal for T cell activation. It is increasingly recognized that programmed death ligand 1 (PD-L1) is a novel co-stimulatory molecule, which raises the question as to whether PD-L1 regulates T cell responses. This study aimed to investigate the inhibitory effects of PD-L1 on T cell activation.Materials and Methods: We constructed a transgenic vector containing the complete PD-L1 gene, which interacts with the inhibitory receptor PD-1 in T cell-mediated immune activation. Donor dendritic cells (DCs) derived from C57BL/6 mice were transfected with PD-L1 and mixed with allogeneic, recipient T cells from BALB/c mice. The T cell activation was determined by the MTT assay and T cell proliferation was determined using carboxyfluoroscein succinimidyl ester (CFSE)-labeling following in vitro mixed leukocyte reactions.Results: The expression of PD-L1 protein in PD-L1-transfected DCs was 47.97% ± 1.06%, compared with 4.66% ± 0.76% and 5.30% ± 0.60% in blank and negative controls, respectively (P < 0.05). PD-L1 protein was effectively expressed in DCs. Furthermore, in DCs stably transfected with PD-L1, T cell activation was significantly suppressed and T cell proliferation rate was decreased by 35% compared with untransfected DCs (P < 0.05).Conclusion: PD-L1 delivers an immunoinhibitory signal, suppressing T cell activation. Overexpression of PD-L1 signaling induces tolerance, which presents a promising immunotherapeutic approach for long-term graft acceptance.

Curcumin Protects Against Sepsis-Induced Acute Lung Injury in Rats - Uncorrected Proof

Xuefei Xiao, Mingshi Yang, Dao Sun, Shenghua Sun — 2011-12-26

The present study aimed to investigate the effect of curcumin on sepsis-induced acute lung injury (ALI) in rats, and explore its possible mechanisms. Male Sprague-Dawley rats were randomly divided into the following five experimental groups (n = 20 per group): animals undergoing a sham cecal ligature puncture (CLP) (sham group); animals undergoing CLP (control group); or animals undergoing CLP and treated with vehicle (vehicle group), curcumin at 50 mg/kg (low-dose curcumin [L-Cur] group), or curcumin at 200 mg/kg (high-dose curcumin [H-Cur] group).At 6, 12, 24 h after CLP, blood, bronchoalveolar lavage fluid (BALF) and lung tissue were collected. The lung wet/dry weight (W/D) ratio, protein level, and the number of inflammatory cells in the BALF were determined. Optical microscopy was performed to examine the pathologic changes in lungs. Myeloperoxidase (MPO) activity, malondialdehyde (MDA) content, as well as superoxidase dismutase (SOD) activity were measured in lung tissues. The expression of inflammatory cytokines, tumor necrosis factor-alpha (TNF-α), interluekin-8 (IL-8), and macrophage migration inhibitory factor (MIF) were determined in the BALF. Survival rates were recorded at 72 h in the five groups in another experiment. Treatment with curcumin significantly attenuated the CLP-induced pulmonary edema and inflammation, as it significantly decreased lung W/D ratio, protein concentration, and the accumulation of the inflammatory cells in the BALF, as well as pulmonary MPO activity. This was supported by the histopathologic examination, which revealed marked attenuation of CLP-induced ALI in curcumin treated rats. In addition, curcumin significantly increased SOD activity with significant decrease in MDA content in the lung. Also, curcumin caused down-regulation of the inflammatory cytokines TNF-α, IL-8, and MIF levels in the lung. Importantly, curcumin improved the survival rate of rats by 40%–50% with CLP-induced ALI. Taken together, these results demonstrate the protective effects of curcumin against the CLP-induced ALI. This effect can be attributed to curcumin ability to counteract the inflammatory cells infiltration and, hence, ROS generation and regulate cytokine effects.

Effective Method to Remove Wound Bacteria: Comparison of Various Debridement Modalities in an In Vivo Porcine Model - Uncorrected Proof

2011-12-26

Background: Debridement is one of the crucial steps for successful wound care. In addition to removing necrotic tissue, debridement has been shown to reduce wound-associated bacteria that delay healing. Using an in vivo porcine model, we compared the effects of various methods of debridement, including hydrosurgery and plasma-mediated bipolar radiofrequency ablation (PBRA), on bacterial removal and wound healing.Methods: One hundred thirty-five deep dermal wounds were inoculated with methicillin resistant Staphylococcus aureus (MRSA) and covered with a polyurethane dressing for 48 h to allow for biofilm formation. Wounds were then treated with either PBRA (at two settings), hydrosurgery, sharp debridement, or no debridement. Biopsies were collected for microbiology and histologic assessment on d 0, 2, 9, and 21 post-treatment.Results: All treatment groups showed a statistically significant reduction in MRSA counts relative to no debridement at all times points (P < 0.05). PBRA at a maximum setting had the lowest MRSA counts at all recovery times and, compared with all other treatment groups, a statistically significant difference was observed on d 21 (P < 0.05). No detrimental effects on the healing process were noted with any of the debridement methods.Conclusion: While sharp debridement has been established as the traditional gold standard for rapid removal of necrotic, infected tissue, our results suggest that novel debridement modalities show clinical promise for the treatment of chronic ulcers and burn wounds, especially when bacteria are present.

Erratum - Uncorrected Proof

2011-12-26

In the March 2011 issue of the Journal of Surgical Research, we regret that in the article, “Trans-Iliac Rat Aorta Stenting: A Novel High Throughput Preclinical Stent Model for Restenosis and Thrombosis,” by Shizu Oyamada, Xiaodong Ma, Tim Wu, Michael P. Robich, Hao Wu, Xingwei Wang, Bryan Buchholz, Stephen McCarthy, Cesario F. Bianchi, Frank W. Sellke, and Roger Laham (J Surg Res 2011;166:e91), the grant information was inadvertently left out. It should have listed in the acknowledgements, “Supported by 5T32-HL007734 (Robich MP).”

Progenitor Cell Mobilizing Treatments Prevent Experimental Transplant Arteriosclerosis - Uncorrected Proof

2011-12-23

Objective: Vascular rejection after organ transplantation is characterized by an arterial occlusive lesion, resulting from intimal proliferation occurring in response to arterial wall immune aggression. Our hypothesis is that an early endothelial repair may prevent vascular graft rejection. The aim of the current study was to compare different pharmacologic progenitor cell mobilizing treatments for their protective effects against vascular rejection.Methods and Results: Aortic transplants were made from balb/c donor to C57Bl/6 recipient mice. Three different mobilizing pharmacologic agents were used: low molecular weight fucoidan (LMWF), simvastatin, and AMD3100. The circulating levels of progenitor cells were found to be increased by all three treatments, as determined by flow cytometry. For each treatment, the design was: treated allografts, nontreated allografts, treated isografts, and nontreated isografts. After 21 d, morphometric and immunohistochemical analyses were performed. We found that the three treatments significantly reduced intimal proliferation, compared with nontreated allografts. This was associated with intimal re-endothelialization of the grafts. Further, in chimeric mice that had previously received GFP-transgenic bone marrow transplantation, GFP-positive cells were found in the vascular allograft intima, indicating that re-endothelialization was, at least partly, due to the recruitment of bone marrow-derived, presumably endothelial progenitor circulating cells.Conclusions: In this aortic allograft model, three different mobilizing treatments were found to partially prevent vascular transplant rejection. Bone marrow-derived progenitor cells mobilized by the three treatments may play a direct role in the endothelial repair process and in the suppression of intimal proliferation.

Projected Lifetime Risks and Hospital Care Expenditure for Traumatic Injury - Uncorrected Proof

Hassan Tetteh — 2011-12-23

Dr. Chang and colleagues present risk and expenditure data for hospitalized trauma patients in California using the cumulative incidence estimate methodology. Their research is important in the context of health care reform, as persistent challenges exist to improve the quality and lower the costs of care. The results and method of cumulative incidence risk presented by Dr. Chang may inform policy makers that negotiate health care's challenges in California and beyond its borders.

Do Primary Neuroendocrine Tumors and Metastasis Have the Same Characteristics? - Uncorrected Proof

2011-12-23

Background: Only one tumor site is usually biopsied to determine the histologic features of that patient’s entire tumor burden. We hypothesized that there are significant histologic and functional differences in primary neuroendocrine tumors (NETS) and their nodal or organ metastases. We also hypothesized that limited tumor sampling could lead to erroneous assumptions about the tumor’s histologic characteristics and clinical behavior.Materials and Methods: Thirteen patients with metastatic well differentiated midgut NETS underwent simultaneous removal of their primary tumor, nodal metastasis, and organ metastasis. Each tumor site was stained quantitatively for Ki-67, chromogranin A (CGA), synaptophysin, CD31, and Factor VIII. Samples were also evaluated with in vitro tumor angiogenesis and drug chemoresistance assays.Results: Ki-67 staining was nearly identical at all sites tested. Quantitative stains for CGA, synaptophysin, cluster of differentiation 31 (CD31), and Factor VIII varied considerably among the patient’s three tissue site samples. Only 6% of the tissue samples tested against a battery of chemotherapeutic agents exhibited susceptibility to a single drug at all three tumor sites. In contrast, several antiangiogenic agents exhibited uniform effectiveness across all three tissue sites in multiple patients.Conclusions: Sampling only one NET tumor site may lead to erroneous assumptions about the tumor’s histologic features and functional behavior. Evaluation of primary tumors and their nodal and organ metastasis may be necessary to optimize clinical decision making.

Hypercoagulability in Porcine Hemorrhagic Shock Is Present Early After Trauma and Resuscitation - Uncorrected Proof

Saad Shebrain, Elizabeth Steensma — 2011-12-23

Despite an increased awareness of the presence and impact of deep venous thrombosis (DVT) and pulmonary embolism (PE), they remain a major source of morbidity and mortality, especially in trauma patients with multisystem injuries. Furthermore, the practice of initiating chemical or even mechanical prophylaxis in this population is greatly limited by their injury and accompanying physiology. Indeed, there is no standardized measurement or definitive algorithm to determine when it is appropriate to initiate DVT prophylaxis, as this remains largely a matter of clinical judgment. Thromboelastography (TEG) is one tool that has been studied in traumatically injured patients as a measure of the degree of coagulopathy, and perhaps of subsequent susceptibility to complications of both hyper- and hypocoagulopathic states.

Systems Biology Approach to Transplant Tolerance: Proof of Concept Experiments Using RNA Interference (RNAi) to Knock Down Hub Genes in Jurkat and HeLa Cells In Vitro1 - Uncorrected Proof

Wint Wah Lwin, Ken Park, Matthew Wauson, Qin Gao, David Perkins, Ajai Khanna — 2011-12-23

Background: Systems biology is gaining importance in studying complex systems such as the functional interconnections of human genes. To investigate the molecular interactions involved in T cell immune responses, we used databases of physical gene-gene interactions to constructed molecular interaction networks (interconnections) with R language algorithms. This helped to identify highly interconnected “hub” genes AT(1)P5C1, IL6ST, PRKCZ, MYC, FOS, JUN, and MAPK1. We hypothesized that suppression of these hub genes in the gene network would result in significant phenotypic effects on T cells and examined this in vitro. The molecular interaction networks were then analyzed and visualized with Cytoscape.Materials and Methods: Jurkat and HeLa cells were transfected with siRNA for the selected hub genes. Cell proliferation was measured using ATP luminescence and BrdU labeling, which were measured 36, 72, and 96 h after activation.Results: Following T cell stimulation, we found a significant decrease in ATP production (P < 0.05) when the hub genes ATP5C1 and PRKCZ were knocked down using siRNA transfection, whereas no difference in ATP production was observed in siRNA transfected HeLa cells. However, HeLa cells showed a significant (P < 0.05) decrease in cell proliferation when the genes MAPK1, IL6ST, ATP5C1, JUN, and FOS were knocked down.Conclusion: In both Jurkat and HeLa cells, targeted gene knockdown using siRNA showed decreased cell proliferation and ATP production in both Jurkat and HeLa cells. However, Jurkat T cells and HELA cells use different hub genes to regulate activation responses. This experiment provides proof of principle of applying siRNA knockdown of T cell hub genes to evaluate their proliferative capacity and ATP production. This novel concept outlines a systems biology approach to identify hub genes for targeted therapeutics.

The Postoperative Serum Interleukin-15 Concentration Correlates with Organ Dysfunction and the Prognosis of Septic Patients Following Emergency Gastrointestinal Surgery - Uncorrected Proof

2011-12-23

Objective: To clarify the time course of changes in the serum interleukin-15 (IL-15) concentrations in septic patients undergoing emergency surgery for abdominal infection and to investigate whether the serum IL-15 levels correlate with the postoperative clinical course of septic patients.Methods: Twenty-four septic patients who had intra-abdominal infection and who underwent an emergency operation were enrolled in this study. The serum IL-15 levels were measured before surgery, and on postoperative d 1 (POD1), POD3, and POD5, and the relationship between the serum IL-15 levels and the postoperative clinical course estimated by the Systemic Inflammatory Response Syndrome (SIRS) criteria, Acute Physiology and Chronic Health Evaluation (APACHE-II) score, the parameters of organ function, and the 30-d mortality were evaluated.Results: The time course of changes of the serum IL-15 levels were significantly different between survivors and non-survivors (P < 0.05, by repeated measures analysis of variance [ANOVA]). There was a statistically significant relationship between the serum IL-15 levels on POD1 or POD3 and the duration of SIRS (R = 0.50, P < 0.05, R = 0.65, P < 0.01, respectively). Furthermore, a significant positive correlation was observed between the serum IL-15 levels on POD1 and the creatinine levels on POD1 or POD3 (R = 0.48, P < 0.05, R = 0.50, P < 0.05, respectively), and a significant negative correlation between the serum IL-15 levels on POD1 or POD 3 and the PaO2/FiO2 on POD3 (R = –0.51, P < 0.05, R = –0.69, P < 0.01, respectively).Conclusions: The measurement of postoperative serum level of IL-15 might be useful for predicting the severity of SIRS and organ dysfunction, especially renal and pulmonary dysfunction.

NBD Peptides Protect Against Ischemia Reperfusion After Orthotopic Liver Transplantation in Rats - Uncorrected Proof

Ming-xiang Cheng, Jian-ping Gong, Yong Chen, Zuo-jin Liu, Bing Tu, Chang-an Liu — 2011-12-23

Background: NBD (NEMO binding domain) peptides could selectively inhibit the inflammation induced NF-κB activity, while sparing the protective functions of basal NF-κB activity. The aim of this study was to determine whether NBD peptides inhibited the transcriptional activity of nuclear factor-κB (NF-κB) during liver transplant ischemia-reperfusion injury (IRI), without affecting its basal function.Materials and Methods: Sprague Dawley (SD) rats were randomly divided into two groups. Orthotropic liver transplantation according to the Kamada technique was performed on 24 pairs of animals. Donors were given NBD peptides (8 mg/kg, intraperitoneal) 2 h before surgery and the controls were treated with the same volume of physiologic saline. An additional 16 animals in normal condition (did not undergo any surgery) were also divided into two groups and given the same treatment as above to assess the effect of NBD peptides on basal function. We analyzed levels of alanine aminotransferase (ALT), tumor necrosis factor (TNF-α), IKK (IκB kinase) complex phosphorylation, IκBα degradation, NF-κB transcriptional activity, apoptosis, and performed a morphologic study of liver tissues at 3, 6, and 24 h after portal vein reperfusion and in normal condition (n = 8).Results: Pretreatment with NBD peptides significantly improved liver function, attenuating liver parenchymal cell damage, apoptosis by down-regulating TNF-α level, inhibiting IKK complex phosphorylation, IκBα degradation, and NF-κB transcriptional activity, but had no effect in normal condition.Conclusion: NBD peptides attenuated hepatic IRI by preventing NF-κB activation, without affecting basal NF-κB activity.

Curcumin for the Prevention of Acute Lung Injury in Sepsis: Is It More Than the Flavor of the Month? - Uncorrected Proof

I. Michael Leitman — 2011-12-21

Curcumin is found in the popular spice turmeric, which is a member of the ginger family. It gives curry its bright yellow color and it is even used as a food color . It has been used for thousands of years as a Chinese and Indian herbal medication . Curcumin has long been known to have anti-inflammatory properties since the 1950s, especially in murine models. Tham and co-workers demonstrated that a curcuminoid substance reduced the lethality in induced abdominal sepsis in mice . The mechanism might be due to up-regulation of peroxisome proliferator-activated receptor (PPAR)-gamma, an anti-inflammatory nuclear receptor. In one study, a bolus injection of curcumin given to rats reduced mortality by this mechanism . It has also been found to have activity in alleviating the chronic changes in asthma . Also, its anti-inflammatory effect appears to be responsible for curcumin's ability to prevent pulmonary toxicity from paraquat . It has also been shown to reduce pulmonary toxicity from bleomycin and oleic acid . Others have also shown the prevention of acute lung injury due to sepsis. Lian and colleagues demonstrated that curcumin reduced endotoxin-mediated lung injury in a rat model . It may also have an effect as an oxygen radical scavenger . In studies examining prevention of radiation-induced lung injury, dietary curcumin was found to ameliorate pulmonary fibrosis and increased mouse survival while not impairing tumor cell killing by radiation . In an article previously published in the Journal of Surgical Research, Liu et al. demonstrated that curcumin helped to prevent cardiopulmonary bypass induced lung injury . Unfortunately, the poor solubility of curcumin, due to its hydrophobic property and preferential interaction with lipid membranes, limits its bioavailability and clinical efficacy. To increase its solubility and bioavailability, Sun and co-workers have demonstrated increased efficacy in preventing lung injury in experimental sepsis in mice through encapsulation in liposomes . Others have shown that injection of Sertoli cells, preloaded with curcumin, reduces the incidence of oxidative injury to the lung and ARDS in a lung transplantation model . Also, on a cellular level, pretreatment with curcumin modulates leukocyte and platelet adhesion improves survival in mice sepsis .

Are There Predictors of Malignancy in Patients with Multinodular Goiter?1 - Uncorrected Proof

Jie Luo, Catherine McManus, Herbert Chen, Rebecca S. Sippel — 2011-12-21

Background: Multinodular goiters (MNG) have recently been shown to have an incidence of cancer that approaches that of solitary thyroid nodule. However, fine needle aspiration (FNA) of a MNG is limited due to the presence of multiple nodules. Therefore we sought to identify risk factors for malignancy in patients with MNG.Methods: A total of 1791 consecutive patients underwent thyroidectomy at a single academic institution between May 1994 and December 2009. Of these, 838 patients had a MNG, which we defined as ≥2 nodules on preoperative ultrasound. The medical records of these patients were reviewed and analyzed.Results: A final pathologic diagnosis of malignancy was found in 260 of 838 (31%) of MNG patients. Of the 260 patients with malignancy, 113 (44%) had a focus of cancer <1 cm. Of the patients with malignancy on final pathology, preoperative FNA detected only 46% (n = 120). Of the 140 cancers not recognized preoperatively, 61 (44%) were >1 cm in size. On univariate analysis risk factors for malignancy were younger age and male gender. Patients with malignant nodules also had smaller nodules, smaller thyroids, and fewer nodules than those patients with benign findings on pathology. On multivariate analysis all predictors remained independently associated with malignancy with the exception of thyroid weight.Conclusion: Risk factors for malignancy in a MNG included male gender, younger age, fewer nodules, and smaller nodule size. The low predictive value of FNA in our population suggests there needs to be better ways to predict malignancy in patients with MNG. Therefore, these clinical risk factors should be considered when consulting patients with MNG in regards to their risk of malignancy.

Phenacetin O-Deethylation is a Useful Tool for Evaluation of Hepatic Functional Reserve in Rats with CCl4-Induced Chronic Liver Injury - Uncorrected Proof

2011-12-21

Background: Mortality of liver resection is as high as 3.1% to 25% in patients with chronic liver disease. Evaluation of hepatic functional reserve is critical for the prediction of risk of postoperation death. Phenacetin O-deethylation is a marker reaction of cytochrome P4501A2 (CYP1A2) activity. In this study, our aim is to investigate whether phenacetin O-deethylation is a useful tool for the evaluation of hepatic functional reserve in rats with chronic liver injury.Materials and Methods: Rat model for chronic liver injury was established by subcutaneous administration of 50% CCl4, 1 mL/kg twice per week for 12 wk. Hepatic CYP1A2 activity, content, and mRNA expression were determined (n = 10). Effects of 15%, 30%, and 45% hepatectomy on phenacetin O-deethylation were evaluated in the rats (n = 5 in each group). Additionally, the correlation of risk of death after 70% hepatectomy with phenacetin O-deethylation was studied in 27 rats with chronic liver injury.Results: Compared with normal controls, CYP1A2 activity, content, and mRNA expression decreased 33%, 60%, and 50% in the rats with chronic liver injury (P < 0.05), respectively. Following the increasing of liver-resected size, CYP1A2 activity decreased proportionally (rs = −0.877, P < 0.05). Six of 27 rats with chronic liver injury died within 7 d after 70% hepatectomy. Phenacetin metabolism was impaired more severely in 6 rats that died than in 21 living rats (P < 0.05).Conclusions: Phenacetin O-deethylation is a useful tool for the evaluation of hepatic functional reserve in the rats with CCl4-induced chronic liver injury.

Bromelain Ameliorates the Wound Microenvironment and Improves the Healing of Firearm Wounds - Uncorrected Proof

Si-Yu Wu, Wei Hu, Bo Zhang, Shuai Liu, Jian-Min Wang, Ai-Min Wang — 2011-12-19

Background: In a previous study, we proposed a new therapy using topical bromelain as a supplement to simple wound-track incision for the debridement of firearm wounds. This enzymatic debridement greatly simplified the management of high-velocity gunshot wounds in a pig model, and bromelain was confirmed to improve wound healing. The purpose of the present study was to investigate the effect of bromelain on the microenvironment of firearm wounds.Methods: Sixteen Chinese landrace pigs wounded by high-velocity projectiles were divided randomly into four groups: wound incision (group I), incision + bromelain (group IB), wound excision (group E), and control. Blood perfusion, oxygen partial pressure (pO2), and the content of tumor necrosis factor (TNF)-α and transforming growth factor (TGF)-β in wound-track tissue were measured. Wound healing was also noted.Results: The recovery of blood perfusion in tissue and pO2 in wound tracks was significantly more rapid in group IB and group E than in group I and control. The tissue level of TNF-α was significantly lower in group IB than in group I and control 48 h and 72 h post-wounding, and was lower than in group E 48 h post-wounding. The tissue level of TGF-β in group IB was sustained at a significantly higher level than in the other three groups. Wound healing time was also shorter in group IB.Conclusions: Enzymatic debridement using topical bromelain in incised wound tracks accelerates the recovery of blood perfusion, pO2 in wound tissue, controls the expression of TNF-α and raises the expression of TGF-β.

Damage Control Resuscitation Decreases Systemic Inflammation After Hemorrhage - Uncorrected Proof

2011-12-19

Background: Severe hemorrhagic shock and resuscitation initiates a dysfunctional systemic inflammatory response leading to end-organ injury. Clinical evidence supports the transfusion of high ratios of plasma and packed red blood cells (pRBCs) in the treatment of hemorrhagic shock. The effects of resuscitation with different ratios of fresh blood products on inflammation and organ injury have not yet been characterized.Materials and Methods: Mice underwent femoral artery cannulation and pressure-controlled hemorrhage for 60 min, then resuscitation with fresh plasma and pRBCs collected from donor mice. Plasma alone, pRBCs alone, and ratios of 2:1, 1:1, and 1:2 plasma:pRBCs were used for resuscitation strategies. Mice were sacrificed to determine biochemical and hematologic parameters, serum cytokine concentrations, tissue myeloperoxidase levels, and vascular permeability.Results: Compared with other resuscitation strategies, mice resuscitated with pRBCs alone exhibited increased hemoglobin levels, while other hematologic and biochemical parameters were not significantly different among groups. Compared with 1:1, mice resuscitated with varying ratios of plasma:pRBCs exhibited increased cytokine concentrations of KC, MIP-1α, and MIP-2, and increased intestinal and lung myeloperoxidase levels. Mice resuscitated with 1:1 had decreased vascular permeability in the intestine and lung as compared with other groups.Conclusions: Resuscitation with a 1:1 ratio of fresh plasma:pRBCs results in decreased systemic inflammation and attenuated organ injury. These findings support the potential advantage of transfusing blood products in physiologic ratios to improve the treatment of severe hemorrhagic shock.

Biologic Scaffold Remodeling in a Dog Model of Complex Musculoskeletal Injury - Uncorrected Proof

Neill J. Turner, John S. Badylak, Douglas J. Weber, Stephen F. Badylak — 2011-12-19

Background: Current treatment principles for muscle injuries with volumetric loss have been largely derived from empirical observations. Differences in severity or anatomic location have determinant effects on the tissue remodeling outcome. Biologic scaffolds composed of extracellular matrix (ECM) have been successfully used to restore vascularized, innervated, and contractile skeletal muscle in animal models but limited anatomic locations have been evaluated. The aim of this study was to determine the ability of a xenogeneic ECM scaffold to restore functional skeletal muscle in a canine model of a complex quadriceps injury involving bone, tendon, and muscle.Materials and Methods: Sixteen dogs were subjected to unilateral resection of the distal third of the vastus lateralis and medial half of the distal third of the vastus medialis muscles including the proximal half of their associated quadriceps tendon. This defect was replaced with a biologic scaffold composed of small intestinal submucosa extracellular matrix (SIS-ECM) and the remodeling response was evaluated at 1, 2, 3, and 6 mo (N = 4 per group).Results: The initial remodeling process followed a similar pattern to other studies of ECM-mediated muscle repair with rapid vascularization and migration of myoblasts into the defect site. However, over time the remodeling response resulted in the formation of dense collagenous tissue with islands of muscle in the segments of the scaffold not in contact with bone, and foci of bone and cartilage in the segments that were adjacent to the underlying bone.Conclusions: SIS-ECM was not successful at restoring functional muscle tissue in this model. However, the results also suggest that SIS-ECM may have potential to promote integration of soft and boney tissues when implanted in close apposition to bone.

Dynamics of Hepatic Gene Expression Profile in a Rat Cecal Ligation and Puncture Model - Uncorrected Proof

2011-12-19

Background: Sepsis remains a major clinical challenge in intensive care units. The difficulty in developing new and more effective treatments for sepsis exemplifies our incomplete understanding of the underlying pathophysiology of it. One of the more widely used rodent models for studying polymicrobial sepsis is cecal ligation and puncture (CLP). While a number of CLP studies investigated the ensuing systemic inflammatory response, they usually focus on a single time point post-CLP and therefore fail to describe the dynamics of the response. Furthermore, previous studies mostly use surgery without infection (herein referred to as sham CLP, SCLP) as a control for the CLP model, however, SCLP represents an aseptic injurious event that also stimulates a systemic inflammatory response. Thus, there is a need to better understand the dynamics and expression patterns of both injury- and sepsis-induced gene expression alterations to identify potential regulatory targets. In this direction, we characterized the response of the liver within the first 24 h in a rat model of SCLP and CLP using a time series of microarray gene expression data.Methods: Rats were randomly divided into three groups: sham, SCLP, and CLP. Rats in SCLP group are subjected to laparotomy, cecal ligation, and puncture while those in CLP group are subjected to the similar procedures without cecal ligation and puncture. Animals were saline resuscitated and sacrificed at defined time points (0, 2, 4, 8, 16, and 24 h). Liver tissues were explanted and analyzed for their gene expression profiles using microarray technology. Unoperated animals (Sham) serve as negative controls. After identifying differentially expressed probesets between sham and SCLP or CLP conditions over time, the concatenated data sets corresponding to these differentially expressed probesets in sham and SCLP or CLP groups were combined and analyzed using a “consensus clustering” approach. Promoters of genes that share common characteristics were extracted and compared with gene batteries comprised of co-expressed genes to identify putatative transcription factors, which could be responsible for the co-regulation of those genes.Results: The SCLP/CLP genes whose expression patterns significantly changed compared with sham over time were identified, clustered, and finally analyzed for pathway enrichment. Our results indicate that both CLP and SCLP triggered the activation of a proinflammatory response, enhanced synthesis of acute-phase proteins, increased metabolism, and tissue damage markers. Genes triggered by CLP, which can be directly linked to bacteria removal functions, were absent in SCLP injury. In addition, genes relevant to oxidative stress induced damage were unique to CLP injury, which may be due to the increased severity of CLP injury versus SCLP injury. Pathway enrichment identified pathways with similar functionality but different dynamics in the two injury models, indicating that the functions controlled by those pathways are under the influence of different transcription factors and regulatory mechanisms. Putatively identified transcription factors, notably including CREB, NF-κB, and STAT, were obtained through analysis of the promoter regions in the SCLP/CLP genes. Our results show that while transcription factors such as NF-κB, HOMF, and GATA were common in both injuries for the IL-6 signaling pathway, there were many other transcription factors associated with that pathway which were unique to CLP, including FKHD, HESF, and IRFF. There were 17 transcription factors that were identified as important in at least two pathways in the CLP injury, but only seven transcription factors with that property in the SCLP injury. This also supports the hypothesis of unique regulatory modules that govern the pathways present in both the CLP and SCLP response.Conclusions: By using microarrays to assess multiple genes in a high throughput manner, we demonstrate that an inflammatory response involving different dynamics and different genes is triggered by SCLP and CLP. From our analysis of the CLP data, the key characteristics of sepsis are a proinflammatory response, which drives hypermetabolism, immune cell activation, and damage from oxidative stress. This contrasts with SCLP, which triggers a modified inflammatory response leading to no immune cell activation, decreased detoxification potential, and hyper metabolism. Many of the identified transcription factors that drive the CLP-induced response are not found in the SCLP group, suggesting that SCLP and CLP induce different types of inflammatory responses via different regulatory pathways.

TGF-α Equalizes Age Disparities in Stem Cell-Mediated Cardioprotection - Uncorrected Proof

2011-12-16

Background: Neonatal mesenchymal stem cells exhibit less cardioprotective potential than their adult counterparts. Transforming growth factor-α (TGF-α) has been shown to stimulate adult stem cell VEGF production, however, it remains unknown whether it may augment neonatal stem cell paracrine function. We hypothesized that TGF-α would equalize adult and neonatal stem cell paracrine function and cardioprotection during acute ischemia/reperfusion.Materials and Methods: Bone marrow mesenchymal stem cells isolated from adult and 2.5 wk-old mice were treated with TGF-α (250 ng/mL) for 24 h. VEGF, HGF, IGF-1, IL-1β, and IL-6 production were measure in vitro, and cells were infused via an intracoronary route using a model of isolated heart perfusion.Results: TGF-α equalized adult and neonatal stem cell VEGF production but did not affect production of HGF, IGF-1, IL-1β, or IL-6. ERK, p38 MAPK, and JNK phosphorylation were greater in adult cells in response to TGF-α. Whereas infusion of adult but not neonatal stem cells was associated with improved myocardial functional recovery during reperfusion, infusions of either TGF-α-pretreated cell group were associated with the greatest functional recovery. TGF-α equalizes adult and neonatal mesenchymal stem cell VEGF production and cardioprotection in association with differential regulation of ERK, p38 MAPK, and JNK phosphorylation.

Impact of Age on Liver Regeneration Response to Injury After Partial Hepatectomy in a Rat Model - Uncorrected Proof

2011-12-16

Background: Liver resection is a feasible treatment for multiple liver diseases. There is no evidence about the impact of age on liver regeneration.Objective: To assess the effect of age on liver regeneration in an experimental in vivo animal model of 70%-partial hepatectomy.Methods: Forty young (Y) and old (O) Wistar male rats (n = 80) were distributed into four groups [controls (C), sham operated (SO), hepatectomy 6 h (H6), and 48 h (H48)]. Different morphometric and biochemical factors, oxidative and nitrosative stress, lipid peroxidation, cytokines kinetics, and histopathologic tissular parameters were determined.Results: Early postoperative mortality was higher in aged rats (P = 0.049). Morphometric determinations, liver regeneration index, and total volume weight were favorable to young rats. Serum transaminase levels were higher in aged rats. Parameters of necrosis (measured by histopathologic injury [HI: 0-I-II-III]), regeneration (measured by bromodeoxyuridine-BrdU incorporation) and apoptosis (determined by the TDT-mediated dUTP nick end labeling-TUNEL) were well-synchronized in young rats. Parameters of oxidative stress such as reduced (GSH), oxidized (GSSG) glutathione and lipid peroxidation (measured by hepatic malondialdehyde –MDA-) were lower in young animals throughout the studied period. Nitrosative stress measured by nitric oxide (NO) end-products was higher in late stages in resected old rats. Pro-inflammatory cytokines (TNF- α) reached higher and earlier levels in aged rats while pro-regenerative cytokines (IL-6) were significantly higher in early stages for young rats and in late stages for aged rats. The levels of TGF-β were higher in young rats.Conclusion: Liver regeneration is delayed and reduced in aged animals submitted to liver resection.

Gender Differences at the Cellular Level - Uncorrected Proof

Robert Allen, Luke Packard Brewster — 2011-12-16

In a paper recently published in the Journal of Surgical Research, Drs. Hogg et al. from the Kibbe laboratory provide a report describing the putative interaction between nitric oxide (NO) therapy and estrogen receptors in vascular smooth muscle cells (VSMCs) isolated and cultured from male and female rodents . Current literature continues to show both systems based and cellular differences due to gender , and the clinical challenge to translate these differences into intelligent treatment strategies makes this work very relevant. We applaud the authors for their work in moving this line of investigation forward by utilizing VSMCs from a variety of origins, including knockout murine cells to define estrogen receptors' impact on proliferation.

Increased JNK in Males Compared with Females in a Rodent Model of Abdominal Aortic Aneurysm - Uncorrected Proof

2011-12-16

Background: In humans, there is a 4:1 male:female ratio in the incidence of abdominal aortic aneurysms (AAAs). c-Jun-N-terminal kinase (JNK) is an important upstream regulator of several enzymes involved in AAA formation, including the matrix metalloproteinases (MMPs). The purpose of this study was to determine if there is a gender difference between males and females in JNK during AAA formation.Materials and Methods: Male and female C57/B6 mice underwent aortic perfusion with elastase or heat inactivated elastase with aortas harvested at d 3 and 14 for phenotype determination, RT-PCR, Western blot, and zymography. Additionally, in vitro experiments using siRNA were conducted to define JNK regulation of matrix metalloproteinases (MMPs). A t-test was used to compare between groups.Results: Males formed larger AAAs at d 14 compared with females (P < 0.001), with significantly higher levels of JNK1 protein, proMMP9, proMMP2, and active MMP2. At d 3, males had more JNK1 mRNA, protein, and MMP activity. Knockdown of JNK 1 or 2 in vitro decreased MMP activity, while knockdown of JNK 1 and 2 together blocked all MMP activity.Conclusion: Alterations in JNK between genders is partially responsible for the differential rates of experimental AAA formation, likely through differential regulation of MMPs.

Effective Oncolytic Vaccinia Therapy for Human Sarcomas - Uncorrected Proof

2011-12-16

Background: Approximately one fourth of bone and soft-tissue sarcomas recur after prior treatment. GLV-1h68 is a recombinant, replication-competent vaccinia virus that has been shown to have oncolytic effects against many human cancer types. We sought to determine whether GLV-1h68 could selectively target and lyse a panel of human bone and soft-tissue sarcoma cell lines in vitro and in vivo.Methods: GLV-1h68 was tested in a panel of four cell lines including: fibrosarcoma HT-1080, osteosarcoma U-2OS, fibrohistiocytoma M-805, and rhabdomyosarcoma HTB-82. Gene expression, infectivity, viral proliferation, and cytotoxicity were characterized in vitro. HT-1080 xenograft flank tumors grown in vivo were injected intratumorally with a single dose of GLV-1h68.Results: All four cell lines supported robust viral transgene expression in vitro. At a multiplicity of infection (MOI) of five, GLV-1h68 was cytotoxic to three cell lines, resulting in >80% cytotoxicity over 7 d. In vivo, a single injection of GLV-1h68 into HT-1080 xenografts exhibited localized intratumoral luciferase activity peaking at d 2–4, with gradual resolution over 8 d and no evidence of spread to normal tissues. Treated animals exhibited near-complete tumor regression over a 28-d period without observed toxicity.Conclusion: GLV-1h68 has potent direct oncolytic effects against human sarcoma in vitro and in vivo. Recombinant vaccinia oncolytic virotherapy could provide a new platform for the treatment of patients with bone and soft tissue sarcomas. Future clinical trials investigating oncolytic vaccinia as a therapy for sarcomas are warranted.

The Association of Manganese Superoxide Dismutase Expression in Barrett's Esophageal Progression With MnTBAP and Curcumin Oil Therapy - Uncorrected Proof

Suzanne C. Schiffman, Yan Li, Robert C.G. Martin — 2011-12-15

Background: The aim of this study was to investigate the relationship between reflux induced bile insult and MnSOD expression, as well as to examine therapies to preserve MnSOD expression. Additionally, we sought to examine the relationship between MnSOD protein expression and MnSOD enzymatic activity.Methods: MnSOD protein expression was determined by Western blot assay and enzymatic activity was determined by SOD assay. The enzymatic activity of the Het-1A and Bar-T cells were compared both before and after treatments.Results: MnSOD expression in Het-1A cells was decreased after bile salt exposure. The cells that received MnTBAP or curcumin oil pretreatment showed increased MnSOD expression compared with control untreated cells. The Bar-T cells showed an increase in MnSOD expression after treatment with bile salts. The cells that were pretreated with MnTBAP displayed a larger increase in MnSOD expression compared with the cells that were not pretreated prior to bile salt exposure. The MnSOD activity was significantly different between the untreated cell lines (P = 0.01) and after treatment with bile salt (P = 0.03). Additionally, Bar-T cells had significantly less MnSOD activity than Het-1A cells after each of the pretreatments.Conclusions: We demonstrated preservation of MnSOD expression in Het-1A cells that were pretreated with antioxidants including MnTBAP, curcumin oil, and certain berry extracts. Additionally, we demonstrated that Bar-T cells have significantly less MnSOD activity than Het-1A cells. These finding have important implications for future studies regarding chemoprevention and the treatment of esophageal cancer.

Effects of Thyroid Hormone Supplementation on Anastomotic Healing After Segmental Colonic Resection - Uncorrected Proof

2011-12-15

Background: Alterations of thyroid hormones in colorectal surgery were previously studied. The aim of the present study was to determine the effects of triiodothyronine (T3) supplementation on anastomotic healing after segmental colectomy.Material and Methods: Thirty male Wistar albino rats were divided into sham (n = 6), control (n = 12), and experimental (n = 12) groups. Sham group rats were immediately sacrificed after segmental colonic resection. Control and experimental group rats underwent resection and anastomosis. Experimental group rats received a single dose of T3 (400 μg/100 g) in postoperative day 1. Half of both control and experimental group rats were sacrificed on postoperative d 3 and the remaining half were sacrificed on postoperative d 7. Hydroxiproline (HP), myeloperoxidase (MPO), thyroid stimulating hormone (TSH), free T3 (FT3), and free thyroxine (FT4) levels, bursting pressure, and histologic analyses of the anastomotic segments were compared.Results: FT3 levels significantly decreased in control groups rats compared with the sham group (P < 0.01). However, T3 hormone given rats had no decline in FT3 levels. Anastomotic bursting pressure was significantly higher in the experimental group rats on postoperative d 7 (P = 0.015). Histopathologic analyses of the anastomotic segments determined significantly more severe edema and necrosis in control group rats (P < 0.05). Collagen deposition in the anastomotic tissue was significantly higher in experimental group rats on postoperative d 7 (P = 0.015).Conclusion: Anastomosis after colon resection is associated with decreased FT3 level. T3 supplementation ameliorates the reduction in FT3 and seems to provide constructive therapeutic effects on anastomotic healing.

Tensile Strength of Biological Fibrin Sealants: A Comparative Study - Uncorrected Proof

2011-12-15

Background: Fibrin sealants are commonly used in liver surgery, although their effectiveness in routine clinical practice remains controversial. Individual sealant characteristics are based on hemostatic effects and adhesion properties that can be experimentally measured using the ‘rat skin test’ or the ‘pig skin test’. This study used a more relevant and realistic experimental canine model to compare the differences in the adhesive properties of four fibrin sealants in hepatectomy: Tisseel/Tissucol, Tachosil, Quixil, and Beriplast.Materials and Methods: A partial hepatectomy was performed in beagle dogs under general anesthesia to obtain liver cross-sections. Fibrin sealants were allocated to dog livers using a Youden square design. The tensile strength measurement was performed using a traction system to measure the rupture stress point of a small wooden cylinder bonded to the liver cross-section.Results: Significantly greater adhesion properties were observed with Tisseel/Tissucol compared with Quixil or Beriplast (P = 0.002 and 0.001, respectively). Similarly, Tachosil demonstrated significantly greater adhesive properties compared with Beriplast (P = 0.009) or Quixil (P = 0.014). No significant differences were observed between Tisseel/Tissucol and Tachosil or between Beriplast and Quixil.Conclusions: The results of this comparative study demonstrate that different fibrin sealants exhibit different adhesive properties. Tisseel/Tissucol and Tachosil provided greatest adhesion to liver cross-section in our canine model of hepatectomy. These results may enable the optimal choice of fibrin sealants for this procedure in clinical practice.

Modified Map-Seeking Circuit: Use of Computer-Aided Detection in Locating Postoperative Retained Foreign Bodies1 - Uncorrected Proof

2011-12-15

Background: More than 98% of intra-operative X-rays taken to search for postoperative retained foreign bodies (RFBs) have negative findings; in over 30% of cases of such X-rays, the finding is a false negative. Newer technologies created to find RFBs must not only reduce the false-negative rate, but also must not increase the burden of detecting RFBs. We have introduced the use of computer-aided detection (CAD) to facilitate the detection of RFBs on X-rays utilizing a modified version of map-seeking circuit (MSC) algorithm the referenced map-seeking circuit (RMSC), for our proof-of-concept study for detection of needles in plain abdominal X-rays.Methods: Images were obtained by using a portable cassette-based X-ray machine and a C-arm (digital) machine, both of which are commonly used in the operating room. The images obtained using these machines were divided into subimages of approximately 250 × 250 pixels each, for a total of 455 subimages from the cassette-based machine (A) and 365 from the digital machine (B) for use as test samples. Images obtained from A and B were analyzed separately using our modified MSC algorithm with a minimum (τ = 0) and a maximum threshold (τ = 0.5).Results: The automated detection rate (positive predictive value) was 86%, with a false positive/negative rate of 10% to 15% when τ was zero.Conclusion: The CAD-based RMSC algorithm has the potential to improve the accuracy with which RFBs can be found in X-rays. Further research is needed to optimize the detection rate and to identify a wider range of RFBs.

Regeneration: Letting the Scaffold do the Work - Uncorrected Proof

Aaron M. Rosado, Luke Packard Brewster — 2011-12-15

Barakat et al,. in a paper recently published in the Journal of Surgical Research, report on the development of a humanized liver segment from a decellularized porcine liver by seeding the “liver” with human fetal hepatocytes and stellate cells The authors demonstrate the ability to remove the cellular contents from the porcine liver and employ a fixation agent to cross-link collagen fibers for scaffold support. The cells were seeded into the scaffold via a perfusion apparatus, and this “liver” segment then demonstrated synthetic function ex vivo and tolerated implantation for 2 h in vivo.

Working Towards a Better Understanding of the Biomechanics of the Carotid Artery - Uncorrected Proof

Robert Alan McCready — 2011-12-12

Perhaps no other arterial bed and its management have been as intensely studied as the carotid artery. Whether to patch or not following carotid endarterectomy (CEA), what type of patch to use, complications associated with patching, and long-term results with the various methods of treatment are still not resolved. Added to the treatment options are carotid artery stents (CAS), which have their own attendant complications. Understanding the biomechanics of the carotid artery is essential in our ongoing efforts to treat carotid artery disease in the most effective way.

Germline Mutations in SMAD4 Disrupt Bone Morphogenetic Protein Signaling - Uncorrected Proof

Jennifer C. Carr, Fadi S. Dahdaleh, Donghong Wang, James R. Howe — 2011-12-02

Introduction: Juvenile polyposis (JP) is an autosomal dominant disease that predisposes to GI malignancies. Germline mutations in the tumor suppressor gene SMAD4 account for approximately 20% of JP cases. SMAD4 is the common intracellular mediator of the TGF-β and bone morphogenetic protein (BMP) pathways. Since mutations in BMP receptor 1A also cause JP, we hypothesize that altered BMP signaling is the underlying defect in JP. We therefore set out to investigate the effect of SMAD4 mutations on BMP signaling.Methods: SMAD4 mutations identified in JP patients were selected for analysis. These were created in SMAD4 pCMV expression vectors (EV) using a PCR-based, site-directed mutagenesis (SDM) approach. SDM clones were confirmed by direct sequencing, then co-transfected with an IdI-BMP Luciferase Responsive Element (BRE-Luc) vector and Renilla control vector into HEK-293T cells. Lysates were then collected after 48 hours, and luciferase activity was quantified using a luminometer. A pCMV empty vector was used as a negative control, and its luciferase activity was considered the baseline for cellular BMP signaling. Results obtained for each SDM clone were compared to those with the wild type (WT) vector. Statistical analysis was performed with the Student's t-test.Results: Eleven distinct mutations from 16 JP patients were analyzed; seven mutations were nonsense, and four were missense. Both type of mutations resulted in reduction of BMP signaling; missense mutations produced an 8–30% reduction in luciferase activity, whereas nonsense mutations led to 30–60% reduction in luciferase activity when compared to the WT clone (). All nonsense mutations led to significantly reduced activity relative to WT (P < 0.05), while the reduction in signaling seen in missense mutations was not statistically significant.Conclusion: SMAD4 germline mutations as seen in the JP patients appear to negatively impact downstream BMP signaling. Nonsense mutations resulted in significantly reduced luciferase activity when compared to missense mutations. These results support the hypothesis that disruption of the BMP signaling pathway is the likely etiology of JP in patients with SMAD4 mutations.

Clinical Significance of Serum Soluble E-cadherin in Colorectal Carcinoma1 - Corrected Proof

2011-12-02

Background: E-cadherin expression has been associated with an outcome in patients with colorectal cancer, and serum levels of soluble E-cadherin (sE-cadherin) are significantly elevated in patients with malignant disease. However, the prognostic value of serum sE-cadherin level has not been demonstrated in colorectal cancer.Methods: Serum samples were collected from 186 patients with colorectal cancer and 21 normal volunteers. Serum sE-cadherin levels were measured using an enzyme-linked immunosorbent assay kit. We investigated the relationship between serum sE-cadherin level and clinicopathologic findings.Results: Mean serum sE-cadherin levels were significantly higher in CRC patients than in controls. Mean sE-cadherin levels were significantly correlated with hepatic metastasis, UICC classification, and poor prognosis. Elevated serum sE-cadherin level was an independent risk factor for predicting poor prognosis, and was an independent marker for predicting hepatic metastasis. Among patients with synchronous hepatic metastases, the prognosis was significantly worse in patients with elevated serum sE-cadherin levels than in those with lower levels.Conclusions: Preoperative elevated sE-cadherin level is associated with poor prognosis in colorectal cancer. Measuring serum sE-cadherin may provide valuable information for predicting prognosis in patients with hepatic metastasis.

Immunocompetent Orthotopic Pancreatic Cancer Murine Model: A Step in the Right Direction - Corrected Proof

Quyen D. Chu, Roger H. Kim — 2011-12-02

Pancreatic cancer is the fourth leading cause of cancer-related deaths in the United States. Despite recent advances in the modern care of this recalcitrant disease, survival rate remains dismal . In the early 1970s, the overall 5-yr survival rate for pancreatic cancer was 3%; this rate now stands at 6% . In addition, pancreatic cancer outcomes have not contributed to the decline in the overall cancer death rates; from 1990 to 2007, the death rate for pancreatic cancer actually had risen by 0.97% in women compared with a decline of 22.9% for prostate cancer and 35.2% for female breast cancer . These sobering statistics only underscore an obvious fact—the need for us to have a better grasp on the biology of the disease. So, where do we begin? For starters, we need an animal model that not only recapitulates human pancreatic cancer but also is feasible so that other investigators can validate our findings. This is paramount if we are to eliminate this deadly disease.

Remodeling of Extracellular Matrix Patch used for Carotid Artery Repair - Uncorrected Proof

Anna Fallon, Traci Goodchild, Ruoya Wang, Robert G. Matheny — 2011-11-28

Background: We evaluated the in vitro strength and in vivo arterial-wall response to an extracellular-matrix-based patch material in a sheep model of carotid artery repair.Materials and Methods: A six-ply sheet of acellular, porcine extracellular matrix (ECM) was subjected to in vitro material strength testing and implanted in 15 sheep for 30, 90, and 180 d. Bovine pericardium was used as a control in some animals. In vivo graft patency was assessed by angiography. Explanted grafts were evaluated by histopathology and burst-strength testing.Results: Mean (SD) in vitro suture retention force of the ECM sheet was 14.5 (3.06) N; tensile strength was 29.7 (6.11) N; and probe burst strength was 185 (22.6) N. In vivo, mild stenosis was observed at 30 d for all patches; stenosis was absent at 90 d in the ECM-repaired arteries but not bovine pericardium controls. Pseudoaneurysm was not observed in any animal. Histopathology showed progressive graft degradation, collagen deposition, formation of neocapillaries and fibrocellular neointima, and endothelialization, but no calcification. Mean (SD) burst pressure for unrepaired arteries was 2608 (858) mmHg and 1473 (694) mmHg for ECM-repaired vessels. Mean change in diameter from unloaded state to burst pressure was 29% (9.7) for unrepaired vessels and 24% (13.4) for ECM-repaired vessels.Conclusions: The six-ply ECM sheet can withstand the forces encountered after carotid artery repair. In sheep, it shows evidence of progressive, constructive remodeling as early as 30 d post-implantation with rapid deposition of endothelium. ECM shows promise as a patch material for CEA repair.

Protective Effects of Melatonin and S-Methylisothiourea on Mechlorethamine Induced Nephrotoxicity - Corrected Proof

2011-11-28

Background: In this study, we aimed to investigate the protective effects of melatonin (MEL) and S-methylisothiourea (SMT) on mechlorethamine (MEC) induced nephrotoxicity.Materials and Methods: A total of 36 male Sprague-Dawley rats were divided into four groups: control, MEC, MEC+MEL, and MEC+SMT. Three groups received single dose of MEC (3.5 mg/kg) via transdermal route. Control animals were given saline only via transdermal route. MEL (100 mg/kg) was administered intraperitoneally 30 min after the application of MEC, and after the same dose of MEL was given every 12 h for a total of six doses. SMT (50 mg/kg) was also given intraperitoneally 30 min after the application of MEC.Results: The tissue TNF-α, IL-1β, and NOx levels were found significantly different for all groups (P < 0.001). MEC application resulted in severe histopathological changes. Melatonin showed meaningful protection against kidney damage. But protection by SMT was weaker. TNF-α and IL-1β levels increased significantly with MEC application, and MEL and SMT ameliorated these increases in kidney tissue. MEC also elevated NOx levels in kidney tissue.Conclusions: Both inflammation and oxidative stress may have an important role in the MEC induced nephrotoxicity. MEL and SMT may also have anti-inflammatory properties, as well as anti-oxidant properties.

Functional Analysis of Alloreactive Memory CD4+ T Cells Derived from Skin Transplantation Recipient and Naïve CD4+ T Cells Derived from Untreated Mice - Corrected Proof

2011-11-28

Background: Memory T cells (TMs) exhibit differential susceptibility to many immunomodulatory strategies that induce immunologic tolerance in naïve T cells, which are believed to be an important barrier to inhibiting rejection and inducing tolerance. As skin grafts are a common model for acquiring TMs, we evaluated function of TMs derived from skin grafts. We also assessed the modulatory effects on memory T cells function of the microRNAs miR-155 and miR-181a, which are involved in regulating cytokine secretion and TCR sensitivity to antigen, respectively.Methods: Memory CD4+ T cells derived from skin-graft recipient mice, and naïve CD4+ T cells from untreated mice, were isolated by negative magnetic selection, and then stimulated with dendritic cells pulsed with donor-specific antigens. Effector function and regulating mechanisms were assessed.Results: In contrast to naïve CD4+ T cells, CD4+ TMs stimulated with donor-specific antigen could quickly generate effector function in terms of proliferation and cytokine secretion; miR-155 and miR-181a levels in CD4+ TMs rapidly increased during immune response compared to naïve CD4+ T cells.Conclusion: Memory CD4+ T cells derived from skin grafts could be used as an experimental tool for evaluating effects of different immune-modulating strategies on TMs. Levels of miR-155 and miR-181a up-regulated quickly in TMs, which could be an important mechanism by which TMs mediate immune responses rapidly.

Berberine Inhibits Lipopolysaccharide- and Polyethylene Particle-Induced Mouse Calvarial Osteolysis In Vivo - Corrected Proof

Xiaoxiao Zhou, Chao Zhang, Xiaoqing Wang, Bingchen An, Peng Zhang, Zhen’an Zhu — 2011-11-24

Background: Wear particle-induced osteolysis could lead to the aseptic loosening of implants. Studies have suggested that endotoxins, such as lipopolysaccharides (LPS), may be the primary causes of wear particle-mediated osteolysis, and that osteolysis may originate from subclinical levels of bacterial infection. However, effective therapies against wear particles and gram-negative bacterial or LPS-induced bone resorption are limited.Materials and Methods: In the current study, the effect of berberine on LPS- and polyethylene (PE) particle-induced osteolysis in vivo was investigated using a mouse calvarial model. Osteoclast number per bone perimeter and eroded surface per bone surface were measured.Results: Berberine (10 mg/kg), injected either simultaneously with LPS or 3 d after LPS (25 mg/kg) treatment, blocked LPS-induced osteoclast recruitment and bone resorption in the mouse calvarial model. A daily single-dose of berberine (10 mg/kg), injected either simultaneously with PE particles or 4 d after treatment with PE particles, blocked PE particle-induced osteoclast recruitment and bone resorption. Berberine treatment markedly decreased LPS and PE particle-induced osteoclast recruitment and bone resorption in the murine calvarial model.Conclusion: These results suggest that berberine may have therapeutic effect for osteolysis induced by wear particles and LPS in gram-negative bacteria.

Activation of Cholinergic Anti-Inflammatory Pathway Contributes to the Protective Effects of 100% Oxygen Inhalation on Zymosan-Induced Generalized Inflammation in Mice - Corrected Proof

2011-11-23

Background: The 100% oxygen inhalation has been demonstrated to have a protective effect on mice with zymosan-induced generalized inflammation. However, the underlying mechanism is largely unknown. The present study was designed to explore the role of the cholinergic anti-inflammatory pathway in this animal model.Methods: Oxygen inhalation was given to mice at 4 and 12 h after zymosan injection. One group of mice underwent vagotomy 7 d before zymosan injection. The other two groups of mice either received nicotinic acetylcholine receptor (nAChR) antagonist mecamylamine, or α7 nicotinic acetylcholine receptor (α7nAChR) antagonist methyllycaconitine 30 min before oxygen was given.Results: The 100% oxygen treatment significantly decreased the serum level of TNF-α and increased the serum level of IL-10. The pathologic changes of the heart, lung, liver, and kidney were attenuated, as well as the dysfunction of liver and kidney. The 7-d survival rate of zymosan-challenged mice was also improved. Conversely, all these protective effects caused by pure oxygen treatment were abolished in those animals that received anti-cholinergic treatments.Conclusions: The cholinergic anti-inflammatory pathway may be involved in the 100% oxygen protective mechanism against zymosan-induced generalized inflammation in mice.

Blockade of Chloride Ion Transport Enhances the Cytocidal Effect of Hypotonic Solution in Gastric Cancer Cells - Corrected Proof

2011-11-23

Background: Cancer cells that are exfoliated into the peritoneal cavity during surgery are viable and have the potential to produce peritoneal recurrence. Although peritoneal lavage with distilled water is applied in some cancer surgeries to kill tumor cells, there is no consensus regarding the optimal methodology and its effects.Methods: Three human gastric cancer cell lines, MKN28, MKN45, and Kato-III, were exposed to distilled water, and the resultant morphologic changes were observed using a microscope. Analysis of cell volume changes was performed using a flow cytometer. To investigate the cytocidal effects of the water, re-incubation of the cells was performed after exposing them to hypotonic solution. Additionally, the effects of 5-nitro-2-3-phenylpropylamino)-benzoic acid (NPPB), a Cl− channel blocker, and R(+)-[(dihydroindenyl)oxy] alkanoic acid (DIOA), a blocker of the K+/Cl− co-transporter, on the cells during their exposure to hypotonic solution were analyzed.Results: After the cells had been exposed to the distilled water, a rapid increase in cell volume occurred followed by cell rupture. In the MKN45 and Kato-III cells, treatment with NPPB increased cell volume by inhibiting regulatory volume decrease and enhanced the cytocidal effects of the hypotonic solution, whereas no such effects were observed in the MKN28 cells. On the other hand, treatment of the MKN28 cells with DIOA inhibited RVD and enhanced the cytocidal effects of hypotonic shock.Conclusion: These findings support the efficacy of peritoneal lavage with distilled water during surgery for gastric cancer and suggest that the regulation of Cl− transport enhances the cytocidal effects of hypotonic shock.

Histologic and Biomechanical Evaluation of Biologic Meshes following Colonization with Pseudomonas aeruginosa - Corrected Proof

2011-11-23

Background: Biologic meshes have become increasingly popular for the repair of abdominal wall defects, especially in contaminated sites. The purpose of this study was to evaluate the histologic and biomechanical properties of biologic mesh in response to a bacterial encounter.Material and Methods: A rat model of Pseudomonas aeruginosa colonization and infection of subcutaneously implanted biologic mesh was used. Samples of biologic meshes [acellular human dermis (ADM) and porcine small intestine submucosa (SIS)] were inoculated with P. aeruginosa (105 or 109 cfu) or saline as a control prior to wound closure (n = 6 per group). After 10 or 20 d, the meshes were harvested. The recovered meshes were analyzed for histologic changes and bacterial recovery as well as the material strength properties. Statistical significance (P < 0.05) was determined using 1-way analysis of variance or Mann-Whitney test.Results: ADM and SIS colonized with 109 cfu P. aeruginosa showed an increased inflammatory response with an associated decrease in neo-vascularization (P < 0.05) at 20 d post-implantation compared with controls. P. aeruginosa had no effect on the tensile strength of ADM, but the tensile strength and modulus of elasticity were reduced for SIS compared with controls at 20 d.Conclusion: Bacterial colonization of ADM and SIS with 109cfu P. aeruginosa negatively effected neovascularization and cellular re-population of the material over time but only SIS showed alterations in their biomechanical properties in response to this gram-negative bacterial challenge.

PKI-587 and Sorafenib Targeting PI3K/AKT/mTOR and Ras/Raf/MAPK Pathways Synergistically Inhibit HCC Cell Proliferation - Corrected Proof

2011-11-23

Background: Deregulated Ras/Raf/MAPK and PI3K/AKT/mTOR signaling pathways are found in hepatocellular carcinoma (HCC). This study aimed to test the inhibitory effects of PKI-587 and sorafenib as single agents or in combination on HCC (Huh7 cell line) proliferation.Materials and Methods: 3H-thymidine incorporation and MTT assay were used to assess Huh7 cell proliferation. Phosphorylation of the key enzymes in the Ras/Raf/MAPK and PI3K/AKT/mTOR pathways was detected by Western blot.Results: We found that PKI-587 is a more potent PI3K/mTOR inhibitor than PI-103. Combination of PKI-587 and sorafenib was a more effective inhibitor of Huh7 proliferation than the combination of PI-103 and sorafenib. Combination of PKI-587 and sorafenib synergistically inhibited epidermal growth factor (EGF)-stimulated Huh7 proliferation compared with monodrug therapy. EGF increased phosphorylation of Ras/Raf downstream signaling proteins MEK and ERK; EGF-stimulated activation was inhibited by sorafenib. However, sorafenib, as a single agent, increased AKT (Ser473) phosphorylation. EGF-stimulated AKT (ser473) activation was inhibited by PKI-587. PKI-587 is a potent inhibitor of AKT (Ser473), mTOR (Ser2448), and S6K (Thr389) phosphorylation; in contrast, rapamycin stimulated mTOR complex 2 substrate AKT(Ser473) phosphorylation although it inhibited mTOR complex 1 substrate S6K phosphorylation. PKI-587, as a single agent, stimulated MEK and ERK phosphorylation. However, when PKI-587 and sorafenib were used in combination, they inhibited all the tested kinases in the Ras/Raf /MAPK and PI3K/AKT/mTOR pathways.Conclusion: The combination of PKI-587 and sorafenib has the advantage over monodrug therapy on inhibition of HCC cell proliferation by blocking both PI3K/AKT/mTOR and Ras/Raf/MAPK signaling pathways.

Ultrasound Visualization of the Spinal Accessory Nerve In Vivo1 - Corrected Proof

Seyed A. Mirjalili, Jill C. Muirhead, Mark D. Stringer — 2011-11-23

Background: Inadvertent injury of the spinal accessory nerve during surgical procedures is a cause of significant morbidity with medicolegal repercussions. Surface anatomy is an unreliable guide to the nerve’s location. We suggest that ultrasound can be used to map the course of the nerve in the posterior triangle of the neck.Materials and Methods: Fifty healthy subjects (28 females, mean age 37 y) were scanned using a VF13-5 linear probe and a Siemens Sonoline Antares ultrasound machine (Siemens Medical Solutions USA Inc., Malvern, PA). The caliber, course, and distribution of the nerve in the posterior triangle of the neck were recorded.Results: The nerve was visualized bilaterally in all subjects, running superficially across the posterior triangle with either a straight (56%) or tortuous (44%) course at a depth of about 3 mm beneath the skin surface. It had a mean caliber of 0.76 ± 0.12 mm. It exited the posterior border of sternocleidomastoid at a mean of 6.7 (4.0–9.4) cm below the mastoid process and 1.1 (0.1–2.1) cm above the great auricular point and penetrated the anterior border of trapezius 5.4 (2.1–9.2) cm above the clavicle. Importantly, 58% of nerves divided into 2–4 branches before penetrating trapezius; the nerve branched on at least one side in 49 of 50 individuals.Conclusions: The spinal accessory nerve and its anatomical variants can be consistently and reliably demonstrated by ultrasound in normal individuals. Surface anatomical landmarks are not a reliable guide to the position and course of the nerve in the posterior triangle. Preoperative mapping of the nerve with ultrasound may reduce the risk of iatrogenic injury.

Elderly Recipients of Hepatitis C Positive Renal Allografts Can Quickly Develop Liver Disease - Uncorrected Proof

2011-11-21

Our institution explored using allografts from donors with Hepatitis C virus (HCV) for elderly renal transplantation (RT). Thirteen HCV– elderly recipients were transplanted with HCV+ allografts (eD+/R–) between January 2003 and April 2009. Ninety HCV– elderly recipients of HCV– allografts (eD–/R–), eight HCV+ recipients of HCV+ allografts (D+/R+) and thirteen HCV+ recipients of HCV– allografts (D–/R+) were also transplanted. Median follow-up was 1.5 (range 0.8–5) years. Seven eD+/R– developed a positive HCV viral load and six had elevated liver transaminases with evidence of hepatitis on biopsy. Overall, eD+/R– survival was 46% while the eD–/R– survival was 85% (P = 0.003). Seven eD+/R– died during follow-up. Causes included multi-organ failure and sepsis (n = 4), cancer (n = 1), failure-to-thrive (n = 1) and surgical complications (n = 1). One eD+/R– died from causes directly related to HCV infection. In conclusion, multiple eD+/R– quickly developed HCV-related liver disease and infections were a frequent cause of morbidity and mortality.

Fluorescence Diagnosis of Metastatic Lymph Nodes Using 5-Aminolevulinic Acid (5-ALA) in a Mouse Model of Colon Cancer - Corrected Proof

Shigeru Kato, Junichiro Kawamura, Kenji Kawada, Suguru Hasegawa, Yoshiharu Sakai — 2011-11-21

Background: Lymph node metastasis is one of the most critical prognostic factors in patients with colorectal cancer. Although regional lymph nodes should be surgically resected and pathologically examined, techniques for the intraoperative diagnosis of lymph node metastasis remain to be well established. Fluorescence diagnosis using 5-aminolevulinic acid (5-ALA) is a promising technique for evaluating various malignancies. After exogenous administration of 5-ALA, protoporphyrin IX (PPIX) accumulates in malignant cells and can be detected as red fluorescence. In this study, we investigated the usefulness of fluorescence diagnosis using 5-ALA for the detection of lymph node metastasis in a mouse model of colon cancer.Materials and Methods: An orthotopic colon cancer model was prepared by inoculating the cecal wall of nude mice with HCA7, a human colon adenocarcinoma cell line. After 3 wk, 40 mg/kg of 5-ALA was administered intraperitoneally (IP) or orally (PO). Fluorescence diagnosis with a D-Light System (Karl Storz) was then performed after 3 or 6 h.Results: In the IP group, PPIX fluorescence was detected in metastatic lymph nodes as well as in other malignant lesions, including primary tumors and abdominal implantations, while non-metastatic nodes were fluorescence-negative. In contrast, no obvious fluorescence was detected in cancerous tissues in the PO group.Conclusions: PPIX fluorescence induced by intraperitoneal injection of 5-ALA allows metastatic lymph nodes to be accurately diagnosed in this mouse model. This technique may facilitate the intraoperative diagnosis of lymph node metastases from colon cancer in a clinical setting.

Antioxidant Effects of Propofol on Tourniquet-Induced Ischemia-Reperfusion Injury: An Experimental Study - Corrected Proof

Fatih Ozkan, Yeşim Şenayli, Huseyin Ozyurt, Unal Erkorkmaz, Bora Bostan — 2011-11-21

Purpose: This experimental study aimed to investigate the antioxidant effects of propofol anesthesia at induction doses in a rat skeletal muscle ischemia/reperfusion injury model.Methods: Twenty-six rats were randomly divided into three groups to receive one of the following interventions: sham operation (n = 6), ischemia/reperfusion (I/R) injury (n = 10), or propofol administration in addition to I/R injury (n = 10). I/R injury was attained by 2-h clamping of femoral artery followed by 3-h perfusion. Then blood and tissue samples were collected for biochemical analysis and histopathologic examination. Glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) enzyme activities and nitric oxide (NO) and malondialdehyde (MDA) levels were measured in both plasma and muscle tissue. In addition, catalase (CAT) activity and protein carbonyl (PC) content were measured in muscle tissue.Results: I/R group had significantly higher SOD activity (9.05 versus 5.63 and 6.18 U/mL, P < 0.05) and NO level (46.77 versus 30.62 and 33.90 μmol/L, P < 0.05) compared with sham-operated group and I/R plus propofol group. In addition, GSH-Px activity of the I/R group was significantly higher than sham-operated group (1.26 versus 1.05 U/mL, P < 0.05). I/R group had significantly higher tissue activities of CAT (0.11 versus 0.06 and 0.04 k/g protein, P < 0.05) and SOD (0.12 versus 0.08 and 0.07 U/mg protein, P < 0.05) compared with the sham and I/R plus propofol group. Histopathologic examination showed that I/R plus propofol group had significantly lower degeneration (P = 0.021) and inflammation (P = 0.028) scores compared with I/R group.Conclusion: Propofol anesthesia seems to enhance the antioxidant capacity against tourniquet induced ischemia-reperfusion injury.

Rapamycin Induces Heme Oxygenase-1 in Liver but Inhibits Bile Flow Recovery after Ischemia - Uncorrected Proof

2011-11-21

Background/Aims: Rapamycin, which is employed in the management of patients undergoing liver surgery, induces the synthesis of heme oxygenase-1 (HO-1) in some non-liver cell types. The aim was to investigate whether rapamycin can induce HO-1 expression in the liver, and to test the effects of rapamycin on liver function in the early phase of ischemia reperfusion (IR) injury.Methods: Isolated rat hepatocytes and a rat model of segmental hepatic ischemia and reperfusion were employed. Bile flow was measured gravimetrically or by using indocyanine green. mRNA and protein (by quantitative PCR and Western blot, respectively) and blood concentrations of rapamycin, bilirubin, and liver marker enzymes were measured.Results: In isolated hepatocytes, rapamycin induced a 6-fold increase in HO-1, comparable to that induced by cobalt proporphyrin (CoPP), and a 2-fold increase in peroxiredoxin-1. Pretreatment of rats with rapamycin resulted in a small increase in liver HO-1 expression, a 20% inhibition of the basal rate of bile flow, and a 50% inhibition in the rate of bile flow recovery after ischemia. CoPP increased basal bile flow by 20% and inhibited bile flow recovery by 50%. These effects were associated with small increases in the blood concentrations of bilirubin and liver marker enzymes.Conclusions: Rapamycin, through HO-1 induction, has the potential to protect the liver against damage in the late phase of IR. The inhibition by rapamycin of bile flow indicates that its actions on liver function in the acute phase of IR injury are complex.

Gastric Emptying is Delayed in Transgastric Notes: A Randomized Study in Swine - Corrected Proof

2011-11-21

Aim: The aim of this study was to evaluate the restoration of gastrointestinal motility after NOTES using capsule endoscopy (CE).Materials and Methods: Twenty adult Yorkshire pigs were randomly assigned to four groups: transgastric NOTES (gNOTES), transrectal NOTES (rNOTES), transvaginal NOTES (vNOTES), and laparoscopy (LAP). At the end of a 30-min peritoneoscopy with identification of seven predetermined organs, an array of eight receivers and the recorder were attached to the abdominal wall. The CE was delivered into the antrum with the help of an endoscope and a polypectomy snare. Animals were kept alive for 14 d.Results: Median time for surgery was longer in gNOTES (56 min, range 47–63) and vNOTES (54 min, range 44–79) than in LAP (32 min, range 32–33; P < 0.05 and P < 0.01) and in rNOTES (45.5 min, range 33–56) (P = ns). This increase was related to a larger incision and longer closure times. Images from the CE were successfully retrieved in 19 cases. The CE was retained in the stomach in all animals in gNOTES (459 min; range 360–600), but only in one animal in rNOTES and vNOTES and in none in the LAP group. Failure of passage of the CE beyond the stomach was associated with gNOTES and longer closure of the incision. Animals in the gNOTES group gained less weight than the others and this change was statistical significant when compared with vNOTES animals (1.7 kg, range –1.98 to 4.5 versus 8.4 kg, range 5.8 to 11.45; P < 0.01).Conclusion: Gastric emptying is delayed after gNOTES peritoneoscopy compared with rNOTES, vNOTES, and LAP and this effect is associated with less weight gain.

The Effect of Remnant Preservation on Patterns of Gene Expression in a Rabbit Model of Anterior Cruciate Ligament Reconstruction - Corrected Proof

Guo ming Xie, Xiao qiao Huang fu, Jin zhong Zhao — 2011-11-21

Background: Anterior cruciate ligament (ACL) reconstruction with remnant preservation technique had been thought to be a more favorable milieu for graft reinnervation, revascularization, and ligamentization. However, the influence of preserving tibial residual fibers on mRNA expression during the graft remodeling process has never been investigated.Materials and Methods: Healthy mature New Zealand white rabbits were randomly assigned to one of four groups: remnant dissected, remnant preserved, sham operated, and normal control. Ligament tissue was dissected at 2, 6, and 12 wk after surgery, and real-time PCR was performed using primers for VEGF, TGF-β1, COLlAl, COL3A1, GAP-43, and NT-3.Results: In the remnant preservation group, mRNA levels for matrix components COL l Al, COL3A1, growth factor TGF-β1, and nerve-related genesGAP-43 all increased 6 wk after surgery, compared with the remnant dissection group (P < 0.05). An increased level of VEGF mRNA was also detected in the remnant preservation group 12 wk after operation (P < 0.05). An increased level of NT-3 mRNA was also observed in the remnant preservation group 2 and 12 wk after operation (P < 0.05).Conclusions: Our results suggest that there is a time dependent alteration of angiogenesis-promoting, repair-related, and nerve-related gene expression after ACL reconstruction during the process of graft remodeling. Furthermore, they demonstrate that remnant preservation in ACL reconstruction determines the different molecular profiles of these target genes, especially during the early stages of graft remodeling, which perhaps explains the potential role in promoting revascularization, reinnervation, and ligamentization.

Mucolysis by Ascorbic Acid and Hydrogen Peroxide on Compact Mucin Secreted in Pseudomyxoma Peritonei - Corrected Proof

Krishna Pillai, Javed Akhter, Terence C. Chua, David L. Morris — 2011-11-21

Background: This study examines the potential efficacy of hydrogen peroxide and ascorbic acid in the dissolution of mucinous ascites from pseudomyxoma peritonei.Methods: The mucolytic action of both ascorbic acid (0%–0.2%) and hydrogen peroxide (0%–3%) are investigated as single agent on mucin samples derived from patient. This was followed by examining the joint action of ascorbic acid (0.2%) and hydrogen peroxide (0%–3.0%) on mucin. To lower the concentration of hydrogen peroxide in the mixture, the action of equal concentration of ascorbic acid/hydrogen peroxide ranging from 0%–0.3% are then examined. Finally, the pH (4.5–7.0) effect on mucolytic properties of equal concentration (0.2%) of ascorbic acid/hydrogen peroxide was studied.Results: At the concentrations examined (0%–0.2%), ascorbic acid showed highest mucolytic activity at 0.2%. Similarly, hydrogen peroxide as a single agent (0%–3.0%) showed highest mucolytic activity at 3.0%. The mucolytic action of hydrogen peroxide (0%–3.0%) containing 0.2% ascorbic acid demonstrated synergistic effects. At equal concentration of the two agents, ranging from 0%–0.5%, maximal mucolytic action was observed at 0.2%. The mucolytic property of the final mixture (0.2% ascorbic acid/0.2% hydrogen peroxide) was pH-dependant and showed maximal degradation at pH 4.5 and declined as it reached towards neutral pH.Conclusion: The current study introduces the potential applicability of a formulation that holds promise as a mucolytic agent in patients with mucinous ascites from pseudomyxoma peritonei.