Endoscopic Image Overlay for the Targeting of Hidden Anatomy in Laparoscopic Visceral Surgery

Limitations associated with the visual information provided to surgeons during laparoscopic surgery increases the difficulty of procedures and thus, reduces clinical indications and increases training time. This work presents a novel augmented reality visualization approach that aims to improve visual data supplied for the targeting of non visible anatomical structures in laparoscopic visceral surgery. The approach aims to facilitate the localisation of hidden structures with minimal damage to surrounding structures and with minimal training requirements. The proposed augmented reality visualization approach incorporates endoscopic images overlaid with virtual 3D models of underlying critical structures in addition to targeting and depth information pertaining to targeted structures. Image overlay was achieved through the implementation of camera calibration techniques and integration of the optically tracked endoscope into an existing image guidance system for liver surgery. The approach was validated in accuracy, clinical integration and targeting experiments. Accuracy of the overlay was found to have a mean value of 3.5 mm ± 1.9 mm and 92.7% of targets within a liver phantom were successfully located laparoscopically by non trained subjects using the approach.

Identification of a highly conserved valine-glycine-phenylalanine amino acid triplet required for HIV-1 Nef function

Background The Nef protein of HIV facilitates virus replication and disease progression in infected patients. This role as pathogenesis factor depends on several genetically separable Nef functions that are mediated by interactions of highly conserved protein-protein interaction motifs with different host cell proteins. By studying the functionality of a series of nef alleles from clinical isolates, we identified a dysfunctional HIV group O Nef in which a highly conserved valine-glycine-phenylalanine (VGF) region, which links a preceding acidic cluster with the following proline-rich motif into an amphipathic surface was deleted. In this study, we aimed to study the functional importance of this VGF region. Results The dysfunctional HIV group O8 nef allele was restored to the consensus sequence, and mutants of canonical (NL4.3, NA-7, SF2) and non-canonical (B2 and C1422) HIV-1 group M nef alleles were generated in which the amino acids of the VGF region were changed into alanines (VGF→AAA) and tested for their capacity to interfere with surface receptor trafficking, signal transduction and enhancement of viral replication and infectivity. We found the VGF motif, and each individual amino acid of this motif, to be critical for downregulation of MHC-I and CXCR4. Moreover, Nef’s association with the cellular p21-activated kinase 2 (PAK2), the resulting deregulation of cofilin and inhibition of host cell actin remodeling, and targeting of Lck kinase to the trans-golgi-network (TGN) were affected as well. Of particular interest, VGF integrity was essential for Nef-mediated enhancement of HIV virion infectivity and HIV replication in peripheral blood lymphocytes. For targeting of Lck kinase to the TGN and viral infectivity, especially the phenylalanine of the triplet was essential. At the molecular level, the VGF motif was required for the physical interaction of the adjacent proline-rich motif with Hck. Conclusion Based on these findings, we propose that this highly conserved three amino acid VGF motif together with the acidic cluster and the proline-rich motif form a previously unrecognized amphipathic surface on Nef. This surface appears to be essential for the majority of Nef functions and thus represents a prime target for the pharmacological inhibition of Nef.

Intraperitoneal mesh implantation for fascial dehiscence and open abdomen

Postoperative fascial dehiscence and open abdomen are severe postoperative complications and are associated with surgical site infections, fistula, and hernia formation at long-term follow-up. This study was designed to investigate whether intraperitoneal implantation of a composite prosthetic mesh is feasible and safe.

Phosphoinositide 3-kinase C2β regulates RhoA and the actin cytoskeleton through an interaction with Dbl

The regulation of cell morphology is a dynamic process under the control of multiple protein complexes acting in a coordinated manner. Phosphoinositide 3-kinases (PI3K) and their lipid products are widely involved in cytoskeletal regulation by interacting with proteins regulating RhoGTPases. Class II PI3K isoforms have been implicated in the regulation of the actin cytoskeleton, although their exact role and mechanism of action remain to be established. In this report, we have identified Dbl, a Rho family guanine nucleotide exchange factor (RhoGEF) as an interaction partner of PI3KC2β. Dbl was co-immunoprecipitated with PI3KC2β in NIH3T3 cells and cancer cell lines. Over-expression of Class II phosphoinositide 3-kinase PI3KC2β in NIH3T3 fibroblasts led to increased stress fibres formation and cell spreading. Accordingly, we found high basal RhoA activity and increased serum response factor (SRF) activation downstream of RhoA upon serum stimulation. In contrast, the dominant-negative form of PI3KC2β strongly reduced cell spreading and stress fibres formation, as well as SRF response. Platelet-derived growth factor (PDGF) stimulation of wild-type PI3KC2β over-expressing NIH3T3 cells strongly increased Rac and c-Jun N-terminal kinase (JNK) activation, but failed to show similar effect in the cells with the dominant-negative enzyme. Interestingly, epidermal growth factor (EGF) and PDGF stimulation led to increased extracellular signal-regulated kinase (Erk) and Akt pathway activation in cells with elevated wild-type PI3KC2β expression. Furthermore, increased expression of PI3KC2β protected NIH3T3 from detachment-dependent death (anoikis) in a RhoA-dependent manner. Taken together, these findings suggest that PI3KC2β modulates the cell morphology and survival through a specific interaction with Dbl and the activation of RhoA.

Proposing new approaches for dating young volcanic eruptions by luminescence methods

The application of luminescence dating to young volcanic sediments has been first investigated over three decades ago, but it was only with the technical innovations of the last decade that such analyses became viable. While current analytical procedures show promise for dating late Quaternary volcanic events, most efforts have been aimed at unconsolidated volcanic tephra. Investigations into direct dating of lava flows or of non-heated volcanoclastics like phreatic explosion layers, however, remain scarce. These volcanic deposits are of common occurrence and represent important chrono- and volcanostratigraphic markers. Their age determination is therefore of great importance in volcanologic, tectonic, geomorphological and climate studies. In this article, we propose the use of phreatic explosion deposits and xenolithic inclusions in lava flows as target materials for luminescence dating applications. The main focus is on the crucial criterion whether it is probable that such materials experience complete luminescence signal resetting during the volcanic event to be dated. This is argued based on the findings from existing literature, model calculations and laboratory tests.

Low-temperature evolution of the Morondava rift basin shoulder in western Madagascar: An apatite fission track study

[1] The evolution of the rift shoulder and the sedimentary sequence of the Morondava basin in western Madagascar was mainly influenced by a Permo-Triassic continental failed rift (Karroo rift), and the early Jurassic separation of Madagascar from Africa. Karroo deposits are restricted to a narrow corridor along the basement-basin contact and parts of this contact feature a steep escarpment. Here, apatite fission track (AFT) analysis of a series of both basement and sediment samples across the escarpment reveals the low-temperature evolution of the exhuming Precambrian basement in the rift basin shoulder and the associated thermal evolution of the sedimentary succession. Seven basement and four Karroo sediment samples yield apparent AFT ages between ∼330 and ∼215 Ma and ∼260 and ∼95 Ma, respectively. Partially annealed fission tracks and thermal modeling indicate post-depositional thermal overprinting of both basement and Karroo sediment. Rocks presently exposed in the rift shoulder indicate temperatures of >60°C associated with this reheating whereby the westernmost sample in the sedimentary plain experienced almost complete resetting of the detrital apatite grains at temperatures of about ∼90–100°C. The younging of AFT ages westward indicates activity of faults, re-activating inherited Precambrian structures during Karroo sedimentation. Furthermore, our data suggest onset of final cooling/exhumation linked to (1) the end of Madagascar's drift southward relative to Africa during the Early Cretaceous, (2) activity of the Marion hot spot and associated Late Cretaceous break-up between Madagascar and India, and (3) the collision of India with Eurasia and subsequent re-organization of spreading systems in the Indian Ocean.

Sodium/hydrogen exchanger NHA2 is critical for insulin secretion in β-cells

NHA2 is a sodium/hydrogen exchanger with unknown physiological function. Here we show that NHA2 is present in rodent and human β-cells, as well as β-cell lines. In vivo, two different strains of NHA2-deficient mice displayed a pathological glucose tolerance with impaired insulin secretion but normal peripheral insulin sensitivity. In vitro, islets of NHA2-deficient and heterozygous mice, NHA2-depleted Min6 cells, or islets treated with an NHA2 inhibitor exhibited reduced sulfonylurea- and secretagogue-induced insulin secretion. The secretory deficit could be rescued by overexpression of a wild-type, but not a functionally dead, NHA2 transporter. NHA2 deficiency did not affect insulin synthesis or maturation and had no impact on basal or glucose-induced intracellular Ca(2+) homeostasis in islets. Subcellular fractionation and imaging studies demonstrated that NHA2 resides in transferrin-positive endosomes and synaptic-like microvesicles but not in insulin-containing large dense core vesicles in β-cells. Loss of NHA2 inhibited clathrin-dependent, but not clathrin-independent, endocytosis in Min6 and primary β-cells, suggesting defective endo-exocytosis coupling as the underlying mechanism for the secretory deficit. Collectively, our in vitro and in vivo studies reveal the sodium/proton exchanger NHA2 as a critical player for insulin secretion in the β-cell. In addition, our study sheds light on the biological function of a member of this recently cloned family of transporters.

Collagen XII interacts with avian tenascin-X through its NC3 domain

Large oligomeric proteins often contain several binding sites for different molecules and can therefore induce formation of larger protein complexes. Collagen XII, a multidomain protein with a small collagenous region, interacts with fibrillar collagens through its C-terminal region. However, no interactions to other extracellular proteins have been identified involving the non-collagenous N-terminal NC3 domain. To further elucidate the components of protein complexes present close to collagen fibrils, different extracellular matrix proteins were tested for interaction in a solid phase assay. Binding to the NC3 domain of collagen XII was found for the avian homologue of tenascin-X that in humans is linked to Ehlers-Danlos disease. The binding was further characterized by surface plasmon resonance spectroscopy and supported by immunohistochemical co-localization in chick and mouse tissue. On the ultrastructural level, detection of collagen XII and tenascin-X by immunogold labeling confirmed this finding.