Lung Isolation in the Patient With a Difficult Airway.

One-lung ventilation is routinely used to facilitate exposure for thoracic surgical procedures and can be achieved via several lung isolation techniques. The optimal method for lung isolation depends on a number of factors that include (1) the indication for lung isolation, (2) anatomic features of the upper and lower airway, (3) availability of equipment and devices, and (4) the anesthesiologist's proficiency and preferences. Though double-lumen endobronchial tubes (DLTs) are most commonly utilized to achieve lung isolation, the use of endobronchial blockers offer advantages in patients with challenging airway anatomy. Anesthesiologists should be familiar with existing alternatives to the DLT for lung isolation and alternative techniques for DLT placement in the patient with a difficult airway. Newer technologies such as videolaryngoscopy with or without adjunctive fiberoptic bronchoscopy may facilitate intubation and lung isolation in difficult airway management.

Biogeochemical Research Priorities for Sustainable Biofuel and Bioenergy Feedstock Production in the Americas.

Rapid expansion in biomass production for biofuels and bioenergy in the Americas is increasing demand on the ecosystem resources required to sustain soil and site productivity. We review the current state of knowledge and highlight gaps in research on biogeochemical processes and ecosystem sustainability related to biomass production. Biomass production systems incrementally remove greater quantities of organic matter, which in turn affects soil organic matter and associated carbon and nutrient storage (and hence long-term soil productivity) and off-site impacts. While these consequences have been extensively studied for some crops and sites, the ongoing and impending impacts of biomass removal require management strategies for ensuring that soil properties and functions are sustained for all combinations of crops, soils, sites, climates, and management systems, and that impacts of biomass management (including off-site impacts) are environmentally acceptable. In a changing global environment, knowledge of cumulative impacts will also become increasingly important. Long-term experiments are essential for key crops, soils, and management systems because short-term results do not necessarily reflect long-term impacts, although improved modeling capability may help to predict these impacts. Identification and validation of soil sustainability indicators for both site prescriptions and spatial applications would better inform commercial and policy decisions. In an increasingly inter-related but constrained global context, researchers should engage across inter-disciplinary, inter-agency, and international lines to better ensure the long-term soil productivity across a range of scales, from site to landscape.

Incidence and risk factors for acute kidney injury after spine surgery using the RIFLE classification.

OBJECT: Earlier definitions of acute renal failure are not sensitive in identifying milder forms of acute kidney injury (AKI). The authors hypothesized that by applying the RIFLE criteria for acute renal failure (Risk of renal dysfunction, Injury to the kidney, Failure of kidney function, Loss of kidney function, and End-stage kidney disease) to thoracic and lumbar spine surgery, there would be a higher incidence of AKI. They also developed a model to predict the postoperative glomerular filtration rate (GFR). METHODS: A hospital data repository was used to identify patients undergoing thoracic and/or lumbar spine surgery over a 5-year period (2006-2011). The lowest GFR in the first week after surgery was used to identify and categorize kidney injury if present. Risk factors were identified and a model was developed to predict postoperative GFR based on the defined risk factors. RESULTS: A total of 726 patients were identified over the study period. The incidence of AKI was 3.9% (n = 28) based on the RIFLE classification with 23 patients in the risk category and 5 in the injury category. No patient was classified into the failure category or required renal replacement therapy. The baseline GFR in the non-AKI and AKI groups was 80 and 79.8 ml/min, respectively. After univariate analysis, only hypertension was associated with postoperative AKI (p = 0.02). A model was developed to predict the postoperative GFR. This model accounted for 64.4% of the variation in the postoperative GFRs (r(2) = 0.644). CONCLUSIONS: The incidence of AKI in spine surgery is higher than previously reported, with all of the patients classified into either the risk or injury RIFLE categories. Because these categories have previously been shown to be associated with poor long-term outcomes, early recognition, management, and follow-up of these patients is important.

Initial responses of rove and ground beetles (Coleoptera, Staphylinidae, Carabidae) to removal of logging residues following clearcut harvesting in the boreal forest of Quebec, Canada.

Increased interest in biomass harvesting for bioenergetic applications has raised questions regarding the potential ecological consequences on forest biodiversity. Here we evaluate the initial changes in the abundance, species richness and community composition of rove (Staphylinidae) and ground beetles (Carabidae), immediately following 1) stem-only harvesting (SOH), in which logging debris (i.e., tree tops and branches) are retained on site, and 2) whole-tree harvesting (WTH), in which stems, tops and branches are removed in mature balsam fir stands in Quebec, Canada. Beetles were collected throughout the summer of 2011, one year following harvesting, using pitfall traps. Overall catch rates were greater in uncut forest (Control) than either stem-only or whole-tree harvested sites. Catch rates in WTH were greater than SOH sites. Uncut stands were characterized primarily by five species: Atheta capsularis, Atheta klagesi, Atheta strigosula, Tachinus fumipennis/frigidus complex (Staphylinidae) and to a lesser extent to Pterostichus punctatissimus(Carabidae). Increased catch rates in WTH sites, where post-harvest biomass was less, were attributable to increased catches of rove beetles Pseudopsis subulata, Quedius labradorensis and to a lesser extent Gabrius brevipennis. We were able to characterize differences in beetle assemblages between harvested and non-harvested plots as well as differences between whole tree (WTH) and stem only (SOH) harvested sites where logging residues had been removed or left following harvest. However, the overall assemblage response was largely a recapitulation of the responses of several abundant species.

The influence of condensed tannin structure on rate of microbial mineralization and reactivity to chemical assays.

We examined how tannin structure influences reactivity in tannin assays and carbon and nitrogen mineralization. Condensed tannins from the foliage of ten tree and shrub species and from pecan shells (Carya illinoensis) had different proportions of: (a) epicatechin (cis) and catechin (trans) isomers, (b) procyanidin (PC) and prodelphinidin (PD) monomers, and (c) different chain lengths. The response of each tannin to several widely used tannin assays was determined. Although there was some variation in response to proanthocyanidin (butanol/HCl) and Folin Ciocalteu assays, we did not deduce any predictable relationship between tannin structure and response to either assay. There was little variation in protein precipitation among the different tannins. To assess biological activity, six of the tannins were incubated with forest humus for 22 days. We determined that, while PC-based tannins remained at least partly extractable for the duration of the incubation, tannins with a high proportion of PD subunits rapidly became unextractable from soil. There was a positive correlation between net nitrogen mineralization and cis chemical structure. Carbon mineralization was enhanced initially by the addition of tannins to humus, but after 22 days, a negative correlation between the proportion of cis subunits and respiration was determined. Overall, we were not able to demonstrate consistent effects of structure on either microbial mineralization or reactivity to chemical assays; such relationships remain elusive.

Cadherin-dependent adhesion of human U373MG glioblastoma cells promotes neurite outgrowth and increases migratory capacity. Laboratory investigation.

OBJECT: The current management of primary CNS tumors involves a multimodal approach, incorporating cytoreductive techniques including resection, radiotherapy, and antiproliferative chemotherapeutic agents. Despite these attempts, the majority of patients with a diagnosis of a high-grade glioma have a dismal prognosis, with the leading cause of treatment failure and tumor recurrence attributable to local invasion of adjacent brain parenchyma. The current study examines the capacity of glioma tumor cells to undergo neurite outgrowth and local migration, specifically focusing on the role of the cadherin cell adhesion system. METHODS: Using a recombinant cadherin ectodomain protein, U373MG human glioblastoma cells were assessed for their ability to adhere and migrate in a cadherin-dependent manner in culture. Adhesion was evaluated via growth assessment and neurite length at 72 hours growth on an immobilized cadherin substrate and compared with other matrix adhesion proteins, such as Type IV collagen and vitronectin. Migratory capacity was measured via modified transwell assays, also using recombinant cadherin ectodomain in comparison with collagen and vitronectin. Results Cadherin adherent cells adopt a fasciculated morphology, with a significant increase in neurite extension, measuring 104 ± 13.3 µm in length, compared with background adhesion on bovine serum albumin and nonfunctional cadherin ectodomain controls measuring 55 ± 4.4 and 47 ± 3.84 µm, respectively (p = 0.029). Significant increases in neurite length compared with controls were also observed in the vitronectin (81 ± 4.69 µm) and Type IV collagen (91 ± 7.7 µm) groups (p = 0.017 and 0.025, respectively). With respect to migration, U373 cells demonstrate increased invasion in response to cadherin ectodomain exposure, whereas vitronectin and Type IV collagen were not potent initiators of migration through the transwell barrier. Both adhesion and migration outcomes were noted in the absence of any relative changes in cell proliferation, indicating a primary role for the cadherin-based adhesion system in tumor invasion. CONCLUSIONS: Cadherin-based adhesion promotes increased adhesion, neurite outgrowth, and migration in human U373MG glioblastoma cells, providing a novel area of research for the development of therapeutic targets addressing local tumor invasion.

The adenosine 2A receptor agonist ATL-146e attenuates experimental posthemorrhagic vasospasm.

OBJECTIVE: Selective adenosine 2A receptor agonists, such as ATL-146e, are known to be potent anti-inflammatory agents devoid of systemic side effects and have been used clinically in a number of disease states. However, adenosine 2A receptor agonists have not been studied in the treatment of cerebral vasospasm after subarachnoid hemorrhage. The present study investigated the efficacy of ATL-146e in the prevention of leukocyte infiltration and attenuation of posthemorrhagic vasospasm. METHODS: The rodent femoral artery model of vasospasm was used. Forty male Sprague-Dawley rats were randomly assigned to four different groups (vehicle, 1 ng/kg/min, 10 ng/kg/min, or 100 ng/kg/min ATL-146e administered via subcutaneous osmotic minipump). Vasospasm was evaluated at posthemorrhage Day 8 (period of peak constriction) by calculating the lumen cross-sectional area (expressed as percent change in luminal area: ratio of blood-exposed vessel to normal saline-exposed vessel) and radial wall thickness. Immunostaining with anti-CD45 monoclonal antibody to detect leukocytes was used to evaluate localized inflammation. RESULTS: Significant vasospasm was noted in the vehicle-treated (blood-exposed) control group (78.5%, P < 0.001; expressed as a ratio of luminal area of the saline [no blood] control), but not in the ATL-146e-treated groups (lumen ratio to control: 105.0, 83.4, and 91.3% for the 1, 10, and 100 ng/kg/min groups, respectively). Additionally, infiltration of inflammatory cells was reduced significantly and radial wall thickness was decreased in the ATL-146e-treated groups compared with the vehicle-treated control group. CONCLUSION: Selective activation of the adenosine 2A receptor with ATL-146e prevented posthemorrhagic vasospasm and reduced leukocyte infiltration in this experimental model. This agent is worthy of further investigation and lends credence to the hypothesis supporting a role for inflammation in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage.

RalA and RalB: antagonistic relatives in cancer cell migration.

The Ral family of small G proteins has been implicated in tumorigenesis, invasion, and metastasis. However, little emphasis has been placed on clarifying the individual roles of the two Ral proteins, RalA and RalB, in these processes in view of their high sequence homology. Here we analyze the separate contributions of RalA and RalB in regulating cell migration, a necessary component of the invasive phenotype, in two human cancer cell lines; UMUC-3, a bladder carcinoma line, and the prostate carcinoma line, DU145. Although inhibiting RalA protein expression by approximately 80% with two different small interfering RNA duplexes had no effect on migration, inhibiting RalB expression to the same extent with two different duplexes resulted in a marked reduction in migration. Inhibiting RalB expression did trigger a significant loss of actin cytoskeleton fibers in UMUC-3 that was not seen with inhibition of RalA expression. Interestingly, simultaneous inhibition of RalA and RalB expression had no effect on migration. However, dual inhibition of RalA and RalB expression in UMUC-3 did result in an almost total loss of actin fibers as well as a reduction in proliferation, particularly in reduced serum conditions. These results suggest that RalA and RalB have different roles in cell migration and that they may in fact act as antagonists with regard to this phenotype. As further verification of this hypothesis, we found that expression of constitutively active RalA inhibited migration, whereas expression of constitutively active RalB stimulated migration, consistent with this model. In summary, we present the first demonstration that despite their significant sequence homology, RalA and RalB have nonoverlapping and opposing functions in cancer cell migration but overlapping functions in cell growth.

Endothelin axis is a target of the lung metastasis suppressor gene RhoGDI2.

Half of patients treated for locally advanced bladder cancer relapse with often fatal metastatic disease to the lung. We have recently shown that reduced expression of the GDP dissociation inhibitor, RhoGDI2, is associated with decreased survival of patients with advanced bladder cancer. However, the effectors by which RhoGDI2 affects metastasis are unknown. Here we use DNA microarrays to identify genes suppressed by RhoGDI2 reconstitution in lung metastatic bladder cancer cell lines. We identify such RNAs and focus only on those that also increase with tumor stage in human bladder cancer samples to discover only clinically relevant targets of RhoGDI2. Levels of endothelin-1 (ET-1), a potent vasoconstrictor, were affected by both RhoGDI2 reconstitution and tumor stage. To test the hypothesis that the endothelin axis is important in lung metastasis, lung metastatic bladder carcinoma cells were injected in mice treated with the endothelin receptor-specific antagonist, atrasentan, thereby blocking engagement of the up-regulated ET-1 ligand with its cognate receptor. Endothelin antagonism resulted in a dramatic reduction of lung metastases, similar to the effect of reexpressing RhoGDI2 in these metastatic cells. Taken together, these experiments show a novel approach of identifying therapeutic targets downstream of metastasis suppressor genes. The data also suggest that blockade of the ET-1 axis may prevent lung metastasis, a new therapeutic concept that warrants clinical evaluation.

Rho proteins in cell migration and metastasis.

The ability of tumors to invade, colonize, and destroy distant organs was first recognized by Jean-Claude Recamier in 1829. He termed this phenomenon metastasis. Since it was first described, metastasis has been shown to be a complex multistep process. One important step is cell migrationa critical component of both cancer-cell invasion at the primary site (allowing cells to gain access to the vasculature) and for cells to penetrate the host tissue at distant sites. The aim of this review is to examine the fundamental role of migration in cancer progression and to discuss the role of the Rho family of small G proteins and their effectors in this process.

The EF-hand Ca2+-binding protein p22 plays a role in microtubule and endoplasmic reticulum organization and dynamics with distinct Ca2+-binding requirements.

We have reported that p22, an N-myristoylated EF-hand Ca(2+)-binding protein, associates with microtubules and plays a role in membrane trafficking. Here, we show that p22 also associates with membranes of the early secretory pathway membranes, in particular endoplasmic reticulum (ER). On binding of Ca(2+), p22's ability to associate with membranes increases in an N-myristoylation-dependent manner, which is suggestive of a nonclassical Ca(2+)-myristoyl switch mechanism. To address the intracellular functions of p22, a digitonin-based "bulk microinjection" assay was developed to load cells with anti-p22, wild-type, or mutant p22 proteins. Antibodies against a p22 peptide induce microtubule depolymerization and ER fragmentation; this antibody-mediated effect is overcome by preincubation with the respective p22 peptide. In contrast, N-myristoylated p22 induces the formation of microtubule bundles, the accumulation of ER structures along the bundles as well as an increase in ER network formation. An N-myristoylated Ca(2+)-binding p22 mutant, which is unable to undergo Ca(2+)-mediated conformational changes, induces microtubule bundling and accumulation of ER structures along the bundles but does not increase ER network formation. Together, these data strongly suggest that p22 modulates the organization and dynamics of microtubule cytoskeleton in a Ca(2+)-independent manner and affects ER network assembly in a Ca(2+)-dependent manner.