Local resection versus radical surgery for parathyroid carcinoma: A National Cancer Database analysis.

INTRODUCTION: Parathyroid carcinoma (PC) is rare and often diagnosed incidentally after local resection (LR) for other indications. Although recommended treatment has traditionally been radical surgery (RS), more recent guidelines suggest that LR alone may be adequate. We sought to further investigate outcomes of RS versus LR for localized PC. MATERIALS AND METHODS: PC patients from 2004 to 2015 with localized disease were identified from the National Cancer Database, then stratified by surgical therapy: LR or RS. Demographic and clinicopathologic data were compared. Cox proportional hazard models were constructed to estimate associations of variables with overall survival (OS). OS was estimated from time of diagnosis using Kaplan-Meier curves. RESULTS: A total of 555 patients were included (LR = 522, RS = 33). The groups were comparable aside from LR patients having higher rates of unknown nodal status (66.9% versus 39.4%; p = 0.003). By multivariable analysis, RS did not have a significant association with OS (hazard ratio (HR) = 0.43, 95% confidence interval (95%CI) = 0.10, 1.83; p = 0.255), nor did positive nodal status (HR = 0.66, 95%CI = 0.09, 5.03; p = 0.692) and unknown nodal status (HR = 1.30, 95%CI = 0.78, 2.17; p = 0.311). There was no difference in OS between the LR and RS groups, with median survival not reached by either group at 10 years (median follow-up = 60.4 months; p = 0.20). CONCLUSIONS: There was no difference in OS between LR and RS for localized PC. RS and nodal status may not impact survival as previously identified, and LR should remain a valid initial surgical approach. Future higher-powered studies are necessary to assess the effects of surgical approaches on morbidity and oncologic outcomes.

BIOACCUMULATION OF MERCURY IN A TERRESTRIAL CARNIVORE, AMERICAN MARTEN (MARTES AMERICANA).

Studies that assess mercury bioaccumulation in small carnivores in terrestrial habitats are limited. We quantified total mercury (THg) in American marten (Martes americana) that were harvested for fur in Michigan, US, during 2013 and 2014. We quantified THg (dry weight) in hair, kidney, and liver samples and further analyzed hair for potential demographic and ecological factors that influence THg bioaccumulation. We found THg concentrations to be the highest in hair (1.228±0.475 µg/g, n=40), followed by kidney (0.922±0.651 µg/g, n=29) and liver (0.344±0.219 µg/g, n=26). Total mercury distributed predictably and significantly between tissue types, and hair was moderately predictive at modeling THg in kidney (R2=0.50, P<0.001, n=29) and weakly predictive in liver (R2=0.35, P<0.001, n=26), suggesting that hair, which is easily obtained, could be a useful sample type for future biomonitoring programs. The concentrations of THg in hair were higher in adults relative to juveniles, and adult female martens had the highest levels of THg (1.980±0.188 µg/g), as compared to juveniles and adult males. Results of generalized linear modeling suggested that THg hair concentrations were positively associated with marten age and trophic position (stable isotope ratio, δ15N). An interaction between δ15N and the year marten carcasses were collected showed that δ15N alone could be highly predictive of THg in some years but not in others. Annual changes in diet could lead to differing rates of mercury bioaccumulation and alter the usefulness of δ15N to predict THg in marten tissues. Further research should explore the connections between changes in prey availability, types of prey consumed, and the influence on bioaccumulation rates of mercury in terrestrial system mesocarnivores.

Pancreaticoduodenectomy and placement of operative enteral access: Better or worse?

BACKGROUND: It is unclear whether placement of operative enteral access (OEA) during pancreaticoduodenectomy (PD) correlates with decreased morbidity. METHODS: A retrospective chart review of patients undergoing PD with and without OEA placement between January 2016 and May 2018 was undertaken. Outcomes included length of stay (LOS), 30- and 90-day readmission, initiation of total parenteral nutrition (TPN), postoperative pancreatic fistula (POPF), delayed gastric emptying (DGE), and surgical site infection (SSI). RESULTS: 69 patients were evaluated; there was a trend toward decreased LOS for patients without OEA (9 vs. 7.5 days, p = 0.07). There were no significant differences in initiation of TPN (9.1% vs 19.4%, p = 0.311), POPF (21.2% vs 11.1%, p = 0.999), DGE (24.2% vs 22.2%, p = 0.999), organ/space SSI (12.1% vs 8.3%, p = 0.702). CONCLUSION: OEA placement at the time of PD is not necessarily associated with improved perioperative morbidity and outcomes, suggesting that OEA may not be necessary and should be considered on a case by case basis. SUMMARY: It is unclear whether placement of operative enteral access (OEA) during pancreaticoduodenectomy (PD) correlates with decreased morbidity. A retrospective review of patients undergoing PD with and without OEA placement between January 2016 and May 2018 was performed, demonstrating that there were no overall significant differences in postoperative complications and outcomes.

Core versus specialty rotations do not affect students' surgical development.

BACKGROUND: Research is scarce on how the diversity of surgical rotations affects students. We sought to assess the effect of core rotations compared to specialty rotations on students' development. METHODS: Students were given a suturing workshop at the beginning of their surgical clerkship along with a questionnaire. They performed both a simple and a complex suturing task at the beginning and end of the 2-month clerkship. The students were divided into 2 groups based on their surgical rotations. Technical skill and exam scores were compared. RESULTS: Thirty-eight students were included in the study. Objective scores increased for the simple task (14.2, standard deviation 4.5 vs 16.4, standard deviation 4.2, P = .04) and the complex task (12.9, standard deviation 5.3 vs 16.5, standard deviation 4.1, P < .01). Times decreased for the simple task (5.1, standard deviation 1.8 vs 4.1, standard deviation 1.3, min, P < .01) and the complex task (7.9, standard deviation 2.7 vs 6.3, standard deviation 1.5, min, P < .01). Using multivariate analysis, we found that reported hours in the operating room per week and previous hands-on experience affected proficiency of the simple suturing task only. Sixteen students had predominantly core surgical rotations. When compared to the 22 students with more specialty rotations, the only difference was gender (87.5% male vs 50.0% male, P = 0.02). There was no significant difference in the completion times (P = .96, .82), the objective scores (P = .06, .120), the written exam scores (P = .57), or the oral exam scores (P = .89). CONCLUSION: In this small study, it was found that the type of students' rotations does not affect surgical skill or knowledge acquisition.

Selection bias: Examining the feasibility, utility, and participant receptivity to incorporating simulation into the general surgery residency selection process.

BACKGROUND: Opportunities exist to revise the current residency selection process to capture desirable candidate competencies. We examined the extent to which components of the American College of Surgeons/Association for Surgical Education simulation-based medical student curriculum combined with a teamwork activity could be used as potential screening method. METHODS: Students participated in a workshop consisting of training/evaluation of knot tying, suturing, airway management, gowning/gloving, and teamwork. Surveys were given to medical students (MS) and faculty/resident/staff (FRS) to examine their opinions about the residency screening process, the most critical competencies to assess, and the effectiveness of each station for candidate evaluation. RESULTS: Communication (FRS, 4.86 ± .35; MS, 4.93 ± .26), leadership (FRS, 4.41 ± .80; MS, 4.5 ± .76), judgment (FRS, 4.62 ± .74; MS, 4.67 ± .62), professionalism (FRS, 4.64 ± .73; MS, 5.00 ± .00), integrity (FRS, 4.71 ± .78; MS, 4.87 ± .35), and grit/resilience (FRS, 4.71 ± .78; MS, 4.53 ± .74) were considered most valuable for candidate screening. The simulation-based curriculum for evaluation of residency candidates was rated lowest by both groups. Open response comments indicated positive perceptions of this process. CONCLUSIONS: Employing simulation to assess candidates may be most beneficial for examining nontechnical attributes. Future work should continue to explore this area.

MicroRNA-101 inhibits growth of epithelial ovarian cancer by relieving chromatin-mediated transcriptional repression of p21(waf¹/cip¹).

PURPOSE: MicroRNA-101 (miR-101) expression is negatively associated with tumor growth and proliferation in several solid epithelial cancers. Enhancer of zeste homolog 2 (EzH2) appears to be a functional target of miR-101. We explore the role of miR-101 and its interaction with EzH2 in epithelial ovarian carcinoma (EOC). METHODS: In situ hybridization (ISH) for miR-101 was performed on EOC patient tissues and normal controls. EOC cell lines were transfected with miR-101 and subjected to growth analysis and clonogenic assays. Cell motility was assessed by Boyden chamber and wound-healing assays. P21(waf1/cip1) and EzH2 interaction was assessed by Chromatin Immunoprecipitation (ChIP) assay in MDAH-2774 cells. SCID mice were assessed for tumor burden after injection with miR-101 or control vector-treated MDAH-2774 cells. RESULTS: ISH analysis revealed a decrease in miR-101 expression in EOC compared with normal tissue. MiR-101 re-expression in EOC cell lines resulted in increased apoptosis, decreased cellular proliferation, invasiveness, and reduced growth of tumor xenografts. CHIP assays revealed that re-expression of miR-101 inhibited the interaction of EzH2 with p21(waf1/cip1) promoter. CONCLUSIONS: MiR-101 re-expression appears to have antitumor effects, providing a better understanding of the role of miR-101 in EOC.

Sulforaphane induces cell cycle arrest by protecting RB-E2F-1 complex in epithelial ovarian cancer cells.

BACKGROUND: Sulforaphane (SFN), an isothiocyanate phytochemical present predominantly in cruciferous vegetables such as brussels sprout and broccoli, is considered a promising chemo-preventive agent against cancer. In-vitro exposure to SFN appears to result in the induction of apoptosis and cell-cycle arrest in a variety of tumor types. However, the molecular mechanisms leading to the inhibition of cell cycle progression by SFN are poorly understood in epithelial ovarian cancer cells (EOC). The aim of this study is to understand the signaling mechanisms through which SFN influences the cell growth and proliferation in EOC. RESULTS: SFN at concentrations of 5-20 microM induced a dose-dependent suppression of growth in cell lines MDAH 2774 and SkOV-3 with an IC50 of ~8 microM after a 3 day exposure. Combination treatment with chemotherapeutic agent, paclitaxel, resulted in additive growth suppression. SFN at ~8 microM decreased growth by 40% and 20% on day 1 in MDAH 2774 and SkOV-3, respectively. Cells treated with cytotoxic concentrations of SFN have reduced cell migration and increased apoptotic cell death via an increase in Bak/Bcl-2 ratio and cleavage of procaspase-9 and poly (ADP-ribose)-polymerase (PARP). Gene expression profile analysis of cell cycle regulated proteins demonstrated increased levels of tumor suppressor retinoblastoma protein (RB) and decreased levels of E2F-1 transcription factor. SFN treatment resulted in G1 cell cycle arrest through down modulation of RB phosphorylation and by protecting the RB-E2F-1 complex. CONCLUSIONS: SFN induces growth arrest and apoptosis in EOC cells. Inhibition of retinoblastoma (RB) phosphorylation and reduction in levels of free E2F-1 appear to play an important role in EOC growth arrest.

Ritonavir blocks AKT signaling, activates apoptosis and inhibits migration and invasion in ovarian cancer cells.

BACKGROUND: Ovarian cancer is the leading cause of mortality from gynecological malignancies, often undetectable in early stages. The difficulty of detecting the disease in its early stages and the propensity of ovarian cancer cells to develop resistance to known chemotherapeutic treatments dramatically decreases the 5-year survival rate. Chemotherapy with paclitaxel after surgery increases median survival only by 2 to 3 years in stage IV disease highlights the need for more effective drugs. The human immunodeficiency virus (HIV) infection is characterized by increased risk of several solid tumors due to its inherent nature of weakening of immune system. Recent observations point to a lower incidence of some cancers in patients treated with protease inhibitor (PI) cocktail treatment known as HAART (Highly Active Anti-Retroviral Therapy). RESULTS: Here we show that ritonavir, a HIV protease inhibitor effectively induced cell cycle arrest and apoptosis in ovarian cell lines MDH-2774 and SKOV-3 in a dose dependent manner. Over a 3 day period with 20 muM ritonavir resulted in the cell death of over 60% for MDAH-2774 compared with 55% in case of SKOV-3 cell line. Ritonavir caused G1 cell cycle arrest of the ovarian cancer cells, mediated by down modulating levels of RB phosphorylation and depleting the G1 cyclins, cyclin-dependent kinase and increasing their inhibitors as determined by gene profile analysis. Interestingly, the treatment of ritonavir decreased the amount of phosphorylated AKT in a dose-dependent manner. Furthermore, inhibition of AKT by specific siRNA synergistically increased the efficacy of the ritonavir-induced apoptosis. These results indicate that the addition of the AKT inhibitor may increase the therapeutic efficacy of ritonavir. CONCLUSION: Our results demonstrate a potential use of ritonavir for ovarian cancer with additive effects in conjunction with conventional chemotherapeutic regimens. Since ritonavir is clinically approved for human use for HIV, drug repositioning for ovarian cancer could accelerate the process of traditional drug development. This would reduce risks, limit the costs and decrease the time needed to bring the drug from bench to bedside.

Telomere maintenance in laser capture microdissection-purified Barrett's adenocarcinoma cells and effect of telomerase inhibition in vivo.

PURPOSE: The aims of this study were to investigate telomere function in normal and Barrett's esophageal adenocarcinoma (BEAC) cells purified by laser capture microdissection and to evaluate the effect of telomerase inhibition in cancer cells in vitro and in vivo. EXPERIMENTAL DESIGN: Epithelial cells were purified from surgically resected esophagi. Telomerase activity was measured by modified telomeric repeat amplification protocol and telomere length was determined by real-time PCR assay. To evaluate the effect of telomerase inhibition, adenocarcinoma cell lines were continuously treated with a specific telomerase inhibitor (GRN163L) and live cell number was determined weekly. Apoptosis was evaluated by Annexin labeling and senescence by beta-galactosidase staining. For in vivo studies, severe combined immunodeficient mice were s.c. inoculated with adenocarcinoma cells and following appearance of palpable tumors, injected i.p. with saline or GRN163L. RESULTS: Telomerase activity was significantly elevated whereas telomeres were shorter in BEAC cells relative to normal esophageal epithelial cells. The treatment of adenocarcinoma cells with telomerase inhibitor, GRN163L, led to loss of telomerase activity, reduction in telomere length, and growth arrest through induction of both the senescence and apoptosis. GRN163L-induced cell death could also be expedited by addition of the chemotherapeutic agents doxorubicin and ritonavir. Finally, the treatment with GRN163L led to a significant reduction in tumor volume in a subcutaneous tumor model. CONCLUSIONS: We show that telomerase activity is significantly elevated whereas telomeres are shorter in BEAC and suppression of telomerase inhibits proliferation of adenocarcinoma cells both in vitro and in vivo.

Lipopolysaccharide up-regulates heat shock protein expression in rat lung pericytes.

BACKGROUND: Heat shock proteins (HSP) function as molecular chaperones, participating in protein folding and maturation throughout the cell. Serum HSPs may correlate with acute lung injury. Pericytes are perivascular cells located abluminally from endothelial cells, and play a regulatory role in capillary leak. It is our hypothesis that pericytes express HSP 60 and HSP 70, and these HSPs are up-regulated in response to lipopolysaccharide (LPS). METHODS: Rat microvascular lung pericytes were isolated and cultured. Cells from passages three to five were used and treated with LPS (control, 10 ng/mL, and 100 ng/mL) for either 4 or 18 h. Immunoblotting and real-time PCR were used to analyze the presence and quantity of HSP 60 and HSP 70. RESULTS: Immunoblotting revealed the presence of HSP 60 and HSP 70 in control pericytes. After 4 h of treatment with LPS (10 ng/mL and 100 ng/mL), no increase in protein expression of HSP 60 or HSP 70 was seen. However, after 18 h an increase in protein expression of HSP 60 and HSP 70 was seen. Real-time PCR demonstrated the presence of HSP 60 mRNA and HSP 70 mRNA in control pericytes. An increase in mRNA was seen after 18 h of LPS treatment, but not after 4 h. CONCLUSIONS: This study provides the first in vitro evidence that rat lung pericytes express HSP 60 and HSP 70. HSP 60 and HSP 70 are up-regulated after 18 h of LPS exposure. Pericyte heat shock protein expression may contribute to the lung's response seen in sepsis.

Lipopolysaccharide activation of pericyte's Toll-like receptor-4 regulates co-culture permeability.

BACKGROUND: Pericytes (PCs) have a synergistic relationship with endothelial cells (MVEC) in regulating capillary permeability. PCs express Toll-like receptor-4 (TLR-4). We hypothesize one mechanism of MVEC/PC co-culture permeability is regulated through lipopolysaccharide (LPS) activation of pericyte TLR-4. METHODS: Rat PCs were harvested and cultured. PCs were transfected with siRNA targeted to TLR-4. Western blotting was used to confirm gene silencing of TLR-4. A previously described co-culture permeability assay was performed after LPS treatment. RESULTS: Western blot confirmed successful silencing of TLR-4 in PCs, which was sustained for 7 days. A dose- and time-dependent effect of LPS on albumin clearance was seen in MVEC/PC co-cultures. Co-cultures with TLR-4 silenced in PCs eliminated the LPS dose-dependent increase in albumin clearance. CONCLUSIONS: TLR-4 regulates pericyte mediated capillary leak seen with LPS exposure. Our TLR-4 silencing model can be used to further investigate TLR-4's role in pericyte mediated capillary leak.

Cytokine production in lipopolysaccharide-exposed rat lung pericytes.

BACKGROUND: Vessels of the pulmonary microvasculature are composed of two cell types: endothelial cells and pericytes. Pericytes are crucial to the development of capillary leak and pulmonary edema seen in acute respiratory distress syndrome (ARDS). Pericytes express toll-like receptor-4, and is upregulated in response to lipopolysaccharide (LPS). The objective of this study was to evaluate secretory cytokine production by rat microvascular pericytes. It is our hypothesis that pericytes secrete interleukin (IL)-1B, IL-6, and tumor necrosis factor (TNF)-A in response to LPS. METHODS: Rat lung pericytes (RLPs) were isolated and grown either alone or in coculture with rat endothelial cells. Cells from passages 3 to 5 were used and treated with LPS (control, 10 ng/mL, and 100 ng/mL) for varying amounts of time. Immunoblotting and reverse transcriptase polymerase chain reaction (RT-PCR) was used for detection and quantification of NF-kB. Enzyme-linked immunosorbent assay and RT-PCR were used for detection and quantification of cytokines. RESULTS: The protein and mRNA for NF-kB was detected in RLPs. Additionally, NF-kB mRNA increased with exposure to LPS. The supernatant of RLPs exposed to LPS contained IL-1B, and IL-1B increased in a time- and dose-dependant manner. An increase in mRNA for IL-1B, IL-6, and TNF-A was seen in a dose-dependant fashion. Cocultures produced significantly less IL-1B when exposed to similar concentrations of LPS. CONCLUSIONS: Pericytes contain the machinery necessary, and produce pro-inflammatory cytokines. Cocultures manufacture less IL-1B then pericytes alone, which is similar to previous coculture observations. Pericyte activation and cytokine production may play a role in capillary leak seen in gram-negative sepsis.

Solid-pseudopapillary tumors of the pancreas: case report and literature review.

Solid pseudopapillary tumors (SPT) of the pancreas are rare neoplasms of low malignant potential that mostly affect young women. These tumors are of unclear pathogenesis, are slow growing, and can become considerably large before causing symptoms. Complete resection is curative in most cases. This is the case of a 39-year-old African-American woman undergoing evaluation for Roux-en-Y gastric bypass, who was found to have a pancreatic mass. Image-guided biopsy revealed SPT. The patient underwent complete excision of the tumor and had an open Roux-en-Y gastric bypass performed concurrently. The patient had an uneventful postoperative course. A review of the literature is presented.

Pericytes and their role in microvasculature homeostasis.

BACKGROUND: The microvascular pericyte was first described in 1873, though it is a cell that has largely been ignored in the clinical literature. Pericytes are multifunctional, polymorphic, perivascular cells that lie within, and contribute to the production of the microvessel basil lamina. MATERIALS: The pericyte is the second cell that comprises the capillary wall, and is in a prime location to be involved with microvascular permeability. The exact sequence of events in Acute Respiratory Distress Syndrome (ARDS) is unknown, though increased permeability (pulmonary edema) is the primary physiological abnormality seen in the early stages. Pericytes are crucial in the development of capillary leak and pulmonary edema seen in ARDS. Pericytes regulate permeability through contractility and apoptosis. RESULTS: Changes in pericyte contractility alter the physical capillary barrier by opening the endothelial junctional space, and are reversible. Pericyte apoptosis leads to a compromise of the barrier effect of the capillary wall, and is a more permanent change. CONCLUSIONS: The purpose of this paper is to review publications of pericyte physiological and pathophysiologic interactions in regards to contractility, apoptosis, and permeability.

Toll-like receptor-4 message is up-regulated in lipopolysaccharide-exposed rat lung pericytes.

BACKGROUND: Pericytes are multifunctional, polymorphic perivascular cells that lie within the microvessel basal lamina, are located on the abluminal side of endothelial cells, and are thought to play a regulatory role in capillary leak observed in sepsis. Toll-Like receptor 4 (TLR-4) has been implicated as the proximal transmembrane receptor for the LPS/CD 14 complex during the activation of lipopolysacharide (LPS)-induced sepsis. It is our hypothesis that TLR-4 is present on lung pericytes and is up-regulated in response to LPS. METHODS: Rat microvascular lung pericytes were isolated and cultured. Cells from passage 3-5 were used and treated with LPS (control, 10 ng/mL, and 100 ng/mL) for 18 h. Immunostaining and immunoblotting were performed to detect the presence of CD-14, TLR-2, and TLR-4. Real-time polymerase chain reaction was used to analyze the presence and quantity of mRNA for CD-14, TLR-2, and TLR-4. RESULTS: Immunostaining and immunoblotting revealed the presence of CD-14, TLR-2, and TLR-4 in pericytes from each treatment group, and real-time polymerase chain reaction confirmed the presence of mRNA for CD-14, TLR-2, and TLR-4. An increase in the mRNA was observed in CD-14, TLR-2, and TLR-4 in the presence of increasing LPS 4 h after treatment. At 18 h after LPS treatment, a decrease in mRNA was noted. CONCLUSIONS: The up-regulation of TLR-4 in the presence of increasing LPS suggests its importance in pericyte LPS-induced activation. Pericyte TLR-4 recognition of LPS could play a role in capillary leak seen in sepsis. These data also demonstrates that pericytes, once thought to be passive participants in the inflammatory cascade, may be active members.

Vascular endothelial growth factor modulates contractile response in microvascular lung pericytes.

BACKGROUND: Pericytes are capillary support cells that may play a role in regulating permeability by their contractile responses. Vascular endothelial growth factor (VEGF) may play a role in the increased permeability found in sepsis and other inflammatory conditions. The purpose of this study was to evaluate the role of VEGF in regulating pericyte contraction. METHODS: Rat microvascular lung pericytes were isolated according to previously described methods and cultured on collagen gel matrices. Cells were exposed to VEGF (10, 100, and 1000 pg/mL) for varying time periods (0, 10, 30, 60, and 120 minutes). The gels were released and their contractile responses digitally quantified. RESULTS: At all doses, VEGF induced initial pericyte relaxation (contraction 85% to 90% of controls; P < .001). This was followed-up by increased and sustained contraction (107% to 120% of controls; P < .01). CONCLUSIONS: VEGF modifies the contractile response of microvascular lung pericytes. This mechanism may play a role in the increased permeability demonstrated in inflammatory states.

TNF-alpha and IL-1beta increase pericyte/endothelial cell co-culture permeability.

BACKGROUND: Pericytes (PC) have a unique synergistic relationship with microvascular endothelial cells (MVEC) in the regulation of capillary permeability. This study investigates the effect of TNF-alpha, IL-1beta, and IL-6 on the microvasculature by measuring changes in PC contractility, and also, albumin permeability across MVEC/PC co-cultures. MATERIALS AND METHODS: Semi-permeable inserts were plated first with rat lung MVEC and then PCs (on the fourth day) at a ratio of 10:1 MVEC/PC. On day 5, 50 ng/ml of TNF-alpha, IL-1beta, and IL-6 were added with or without a secretory phospholipase A(2)-IIA (sPLA(2)-IIA) inhibitor for 24 h. After treatments, albumin clearances were quantified. For measuring contractility, PCs were cultured on collagen matrices and exposed for 24 h to TNF-alpha, IL-1beta, and IL-6 at 1 ng/ml, 10 ng/ml, and 50 ng/ml with/without inhibitors for sPLA(2)-IIA, phospholipase A(2) (PLA(2)), and cyclooxygenase-II (COX-II). After treatments, the surface area of the collagen disks was digitally quantified. RESULTS: TNF-alpha and IL-1beta significantly increased albumin clearance in MVEC/PC co-cultures (P < 0.05) and induced dose-dependent relaxation of PCs (P < 0.05). PC relaxation was completely attenuated with the sPLA(2)-IIA and pLA(2) inhibitors; the COX-II inhibitor provided partial blockade. IL-6 had no effect on PC contractility or permeability. CONCLUSION: TNF-alpha and IL-1beta directly increased microvascular permeability in co-cultures. They also induced relaxation of PCs through a sPLA(2)-IIA dependent mechanism. Interestingly, IL-6 had no effect, although its presence in high levels has been demonstrated in inflamed lungs. These findings may help elucidate the significance of PC in regulating the capillary response to various pro-inflammatory cytokines.

Long-term follow up and prognostic factors for cryotherapy of malignant liver tumors.

BACKGROUND: The aims of this study were to determine the long-term outcomes of cryotherapy in patients with hepatic malignancies and to describe prognostic factors that may affect survival. METHODS: Ninety-eight patients (56 colorectal metastases, 28 noncolorectal metastases, 14 hepatocellular carcinomas) undergoing hepatic cryosurgery were identified in a retrospective review from January 1994 to December 2002. RESULTS: Overall survival rates at 1-, 2-, 3-, and 5- years were 81%, 62%, 48%, and 28%, respectively(median survival, 33 months) compared to a hepatic recurrence-free survival of 76%, 42%, 24%, and 16%, respectively (median hepatic recurrence-free survival, 20 months). Median follow-up was 54 months. Three hundred lesions were cryoablated; the recurrence per cryolesion was 5%. Major complications were the lone factor that significantly reduced overall (P=.0005) and hepatic recurrence-free survival (P=.0005). The number of lesions (TNL) and total estimated area (TEA) cryoablated did not significantly affect overall or hepatic recurrence-free survival. Additionally, outcomes depending on tumor type were not significantly different. CONCLUSIONS: Cryotherapy is an important option for a wide range of unresectable malignant hepatic tumors and provides the potential for long-term survival. Patients with major complications at the time of cryotherapy suffer a decreased overall and hepatic recurrence-free survival.

Endothelial cells protect against lipopolysaccharide-induced caspase-3-mediated pericyte apoptosis in a coculture system.

BACKGROUND: Cells that comprise the pulmonary capillary walls, the pericytes and endothelial cells, may undergo apoptosis in inflammatory states. This study examined the effects of lipopolysaccharide (LPS) on apoptosis in pericytes and endothelial cells, both individually and grown together in a coculture system. METHODS: Pericytes and endothelial cells were isolated and cultured separately and in coculture as previously described. The cells were subsequently exposed to LPS for 12, 24, 48, and 72 hours. The cellular contents were then examined by Western blot analysis for products of apoptosis. TUNEL staining was also performed to analyze for apoptosis. RESULTS: Pericytes alone exposed to LPS showed increased levels of p11 and p17, which are activated fragments of capase-3, a cysteine effector protease involved in cleaving cytoskeletal and nuclear proteins to induce apoptosis. When grown in coculture with endothelial cells and exposed to LPS in coculture but harvested independently, pericytes showed decreased levels of p11 and p17 and increased levels of Bcl-xL, an antiapoptotic protein that protects the integrity of mitochondria, and prevents cytochrome c release and subsequent caspase-9 activation. CONCLUSIONS: In response to LPS, pericytes undergo apoptosis involving the caspase-3 pathway. Endothelial cells may decrease this effect through the expression of a soluble mediator.

Prognostic determinants in duodenal injuries.

A retrospective review of 222 consecutive patients with duodenal injuries admitted to an urban Level 1 Trauma Center who subsequently underwent laparotomy during the period July 1980 to April 2002 was performed in an effort to elucidate factors associated with mortality, infectious morbidity, and length of stay in these patients. Predictably, the patients were predominantly male (92.7%) and young (mean age, 31.6 years). The overall mortality rate was 22.5 per cent, with a mortality rate of 18 per cent seen in the first 48 hours. Penetrating trauma was suffered by 88.3 per cent of the patients. Multivariate analysis revealed the performance of a thoracotomy, initial emergency department (ED) systolic blood pressure (SBP) <90 mm Hg, final operating room (OR) core body temperature less than 35 degrees C, and presence of a splenic injury to be the most important predictors of mortality (all P < 0.05). Mortality in the patients undergoing a resuscitative thoracotomy was 88.9 per cent versus 13.3 per cent in those patients not requiring thoracotomy. An initial SBP in the ED <90 was associated with a 46 per cent mortality rate, as compared with 4 per cent in those patients not in shock. A final OR core body temperature of less than 35 degrees C led to a 60 per cent mortality rate versus 8.3 per cent for warmer patients. Patients with a concomitant splenic injury were noted to have a 62.5 per cent mortality rate; those without had a 19.4 per cent mortality rate. The mean length of stay among survivors greater than 48 hours was 16.0 +/- 24.7 days. Univariate analyses revealed lowest OR core body temperature below 35 degrees C, initial OR SBP <90, presence of infection, >5 transfusions, initial ED SBP <90, final OR core temperature <35 degrees C, colon injury, spleen injury, and an injury severity score (ISS) >25 all to be significantly associated with increased length of stay. Multivariate analysis revealed an initial operating room blood pressure of less than 90 mm Hg systolic, the presence of an infection, and greater than 5 blood transfusions to be the factors most significantly correlated with increased length of stay (all P < 0.02). Of 182 patients surviving 48 hours, 98 (54%) developed an infection. Fifty-seven (31%) patients were noted to have wound-related infections, 92 (51%) patients had nosocomial infections, and 50 (27%) patients had both types. The presence of an abdominal arterial injury, an ISS >25, pancreatic injury, and lowest OR core body temperature <35 degrees C were the factors identified on multivariate analysis most significantly correlated with infectious morbidity (all P < 0.05). This data suggests that early efforts to prevent shock and rapidly control bleeding are the most likely efforts to reduce mortality rates in these patients. Those patients with duodenal injury presenting in shock or requiring a thoracotomy for resuscitation did poorly. Splenic injury was the associated injury found on multivariate analysis to be most closely associated with increased mortality. Early control of bleeding and the prevention of infection provide the most significant opportunity for decreasing length of stay. Infections are common with duodenal injuries, and aggressive surveillance should especially be performed in those patients with an abdominal arterial injury, an ISS >25, pancreatic injury, or lowest OR core body temperature <35 degrees C.