Germline transformation of the western corn rootworm, Diabrotica virgifera virgifera.

The western corn rootworm (WCR), a major pest of maize, is notorious for rapidly adapting biochemically, behaviourally and developmentally to a variety of control methods. Despite much effort, the genetic basis of WCR adaptation remains a mystery. Since transformation-based applications such as transposon tagging and enhancer trapping have facilitated genetic dissection of model species such as Drosophila melanogaster, we developed a germline-transformation system for WCR in an effort to gain a greater understanding of the basic biology of this economically important insect. Here we report the use of a fluorescent-marked Minos element to create transgenic WCR. We demonstrate that the transgenic strains express both an eye-specific fluorescent marker and piggyBac transposase. We identified insertion-site junction sequences via inverse PCR and assessed insertion copy number using digital droplet PCR (ddPCR). Interestingly, most WCR identified as transgenic via visual screening for DsRed fluorescence proved to carry multiple Minos insertions when tested via ddPCR. A total of eight unique insertion strains were created by outcrossing the initial transgenic strains to nontransgenic WCR mates. Establishing transgenic technologies for this beetle is the first step towards bringing a wide range of transformation-based tools to bear on understanding WCR biology.

Interactions and competition processes among tree species in young experimental mixed forests, assessed with chlorophyll fluorescence and leaf morphology.

Chlorophyll a fluorescence (ChlF) and leaf morphology were assessed in two sites in Europe (Kaltenborn, Germany, and Satakunta, Finland) within a forest diversity experiment. Trees at Satakunta, planted in 1999, form a stratified canopy, while in Kaltenborn the trees are 7 years old, with no apparent canopy connection among broadleaf species. The following ChlF parameters from measured OJIP transient curves were examined: F(V)/F(M) (a proxy for maximum quantum yield); ΨEo (a proxy for efficiency in transferring an electron from reduced QA to the electron transport chain); I-P phase (a proxy for efficiency of reducing final acceptors beyond PSI); and PItot (total performance index for potential energy conservation from photons absorbed by PSII to reduction of PSI end acceptors). At Satakunta F(V)/F(M) and ΨEo in Betula pendula were higher in monocultures and lower in mixed plots, perhaps due to increasing light availability in mixed plots, which can induce photoinhibition. The opposite trend was observed in Picea abies, which was shaded in mixed plots. At Kaltenborn F(V)/F(M) decreased in Fagus sylvatica and P. abies in mixed plots due to competition both above- and belowground. At Satakunta LMA increased in B. pendula leaves with increasing species richness. Leaf area of ten leaves was reduced in F. sylvatica in mixed plots at Kaltenborn. By up-scaling the overall fluorescence response to plot level (PItot_plot ), a significant positive correlation with tree diversity was found at Kaltenborn, but not at Satakunta. This could suggest that competition/facilitation processes in mixed stands play a significant role in the early stages of forest establishment, but then tend to be compensated in more mature stands.

Postoperative monitoring of distal intraarticular radial fractures treated with osteosynthesis by means of multislice CT.

PURPOSE: This study evaluates multislice CT (MSCT) for the postsurgical control of intraarticular fractures of the distal radius. MATERIALS AND METHODS: In 131 patients conventional X-rays in two planes and MSCT were performed. In a consensus process two experienced radiologists judged conventional X-ray and computed tomography images in a random view. The correct position of the osteosynthetic material was assessed and it was ascertained whether an articular surface incongruity, a gap between two fracture fragments, or an intraarticular bone element was detectable. For each point of evaluation a confidence level of the evaluation was assessed on a five-point scale. RESULTS: Ninety patients were classified as having correct alignment and osteosynthesis according to conventional X-ray, while when using the findings from axial CT scans only, 82 patients, and after the addition of multiplanar re-constructions (MPR) only 73 patients were found to have almost proper alignment and osteosynthesis. In 42 patients dehiscence of the fragments was diagnosed with MSCT, but was not visualized by X-ray, leading to surgical revision in eight patients, confirming the diagnosis. In five patients an intraarticular position of intraarticular material was confirmed with MSCT and surgical revision. A significant advantage for the evaluation confidence level was achieved for MPRs in comparison with axial CT scans and X-ray. CONCLUSION: The marked diagnostic advantage with a high evaluation confidence level in comparison with conventional X-ray methods justify the use of MSCT during postoperative monitoring of articular radial fractures treated with osteosynthesis.

General stabilizing effects of plant diversity on grassland productivity through population asynchrony and overyielding.

Insurance effects of biodiversity can stabilize the functioning of multispecies ecosystems against environmental variability when differential species' responses lead to asynchronous population dynamics. When responses are not perfectly positively correlated, declines in some populations are compensated by increases in others, smoothing variability in ecosystem productivity. This variance reduction effect of biodiversity is analogous to the risk-spreading benefits of diverse investment portfolios in financial markets. We use data from the BIODEPTH network of grassland biodiversity experiments to perform a general test for stabilizing effects of plant diversity on the temporal variability of individual species, functional groups, and aggregate communities. We tested three potential mechanisms: reduction of temporal variability through population asynchrony; enhancement of long-term average performance through positive selection effects; and increases in the temporal mean due to overyielding. Our results support a stabilizing effect of diversity on the temporal variability of grassland aboveground annual net primary production through two mechanisms. Two-species communities with greater population asynchrony were more stable in their average production over time due to compensatory fluctuations. Overyielding also stabilized productivity by increasing levels of average biomass production relative to temporal variability. However, there was no evidence for a performance-enhancing effect on the temporal mean through positive selection effects. In combination with previous work, our results suggest that stabilizing effects of diversity on community productivity through population asynchrony and overyielding appear to be general in grassland ecosystems.

piggyBac-based insertional mutagenesis in Tribolium castaneum using donor/helper hybrids.

We describe an efficient method for generating new piggyBac insertions in the germline of F(1) hybrid Tribolium castaneum derived from crosses between transgenic helper and donor strains. Helper strains carried single Minos elements encoding piggyBac transposase. The donor strain carried a single piggyBac element inserted into an actin gene, expanding the eye-specific, 3xP3-EGFP (enhanced green fluorescent protein) reporter expression domain to include muscle. Remobilization of the donor element is accompanied by loss of muscle fluorescence but retention of eye fluorescence. In a pilot screen, the piggyBac donor was remobilized in 84% of the hybrid crosses, generating hundreds of new lethal, enhancer-trap, semisterile and other insertions. The jumpstarter system described herein makes genome-wide, saturation insertional mutagenesis a realistic goal in this coleopteran species.

[Clinical relevance of multislice CT of the spine after osteosynthesis]. / Postoperative Kontrolle von osteosynthetischen Versorgungen der Wirbelsäule mit der Mehrzeilen-Spiral-CT.

PURPOSE: To examine the clinical relevance of multislice CT (MSCT) scans in postoperative checks of the spine after osteosynthesis. MATERIAL AND METHODS: The results of x-rays (apical lateral) in 30 patients having spinal surgery involving osteosynthesis (5 x metastasis, 20 x fractures, 3 x spondylolisthesis, 2 x scoliosis) were correlated to those of MSCT (140 kV, 200 mAs, collimation 4 x 1 mm, pitch 0.75; VolumeZoom, Siemens, Erlangen, Germany) with multiplanar reconstruction. Two radiologists independently checked the conventional x-ray and multislice CT scans for anatomical positioning, damage of osteosynthetic material, and intraspinal dislocation of bone fragments. RESULTS: By correlating conventional spinal x-rays with MSCT, additional diagnostic information was gained. In 9 of 30 patients MSCT revealed the extraosseal location of a screw tip (x-ray, 5/30), in 8/30 patients a narrowing of the spinal canal by osteosynthetic material was detected in MSCT (x-ray, 4/30), in 2/30 patients osteal fragments were detected in the vertebral canal by MSCT (x-ray, 0/30). In MSCT and in conventional x-ray a fracture of the osteosynthesis was correctly diagnosed in 3 patients. One patient underwent corrective surgery for dislocated osteosynthetic material, which was solely diagnosed with MSCT. CONCLUSION: Due to the high degree of additional diagnostic information MSCT seems to be the method of choice for postoperative spinal surgery involving osteosynthesis.

The Tribolium chitin synthase genes TcCHS1 and TcCHS2 are specialized for synthesis of epidermal cuticle and midgut peritrophic matrix.

Functional analysis of the two chitin synthase genes, TcCHS1 and TcCHS2, in the red flour beetle, Tribolium castaneum, revealed unique and complementary roles for each gene. TcCHS1-specific RNA interference (RNAi) disrupted all three types of moult (larval-larval, larval-pupal and pupal-adult) and greatly reduced whole-body chitin content. Exon-specific RNAi showed that splice variant 8a of TcCHS1 was required for both the larval-pupal and pupal-adult moults, whereas splice variant 8b was required only for the latter. TcCHS2-specific RNAi had no effect on metamorphosis or on total body chitin content. However, RNAi-mediated down-regulation of TcCHS2, but not TcCHS1, led to cessation of feeding, a dramatic shrinkage in larval size and reduced chitin content in the midgut.

[Dose optimization for multislice computed tomography protocols of the midface]. / Dosisoptimierung der Mehrzeilen-Spiral-CT (MSCT) des Mittelgesichts.

PURPOSE: To optimize multislice computed tomography (MSCT) protocols of the midface for dose reduction and adequate image quality. MATERIALS AND METHODS: MSCT (Somatom Volume Zoom, Siemens) of the midface was performed on 3 cadavers within 24 hours of death with successive reduction of the tube current, applying 150, 100, 70 and 30 mAs at 120 kV as well as 40 and 21 mAs at 80 kV. At 120 kV, a pitch of 0.875 and collimation of 4x1 mm were used, and at 80 kV, a pitch of 0.7 and collimation of 2x0.5 mm. Images were reconstructed in transverse and coronal orientation. Qualitative image analysis was separately performed by two radiologists using a five-point scale (1 = excellent; 5 = poor) applying the following parameters: image quality, demarcation and sharpness of lamellar bone, overall image quality, and image noise (1 = minor; 5 = strong). The effective body dose [mSv] and organ dose [mSv] of the ocular lens (using the dosimetry system "WINdose") were calculated, and the interobserver agreement (kappa coefficient) was determined. RESULTS: For the evaluation of the lamellar bone, adequate sharpness, demarcation and image quality was demonstrated at 120 kV/30 mAs, and for the overall image quality and noise, 120 kV/40 mAs was acceptable. With regard to image quality, the effective body dose could be reduced from 1.89 mSv to 0.34 mSv and the organ dose of the ocular lens from 27.2 mSv to 4.8 mSv. Interobserver agreement was moderate (kappa = 0.39). CONCLUSION: Adequate image quality was achieved for MSCT protocols of the midface with 30 mAs at 120 kV, resulting in a dose reduction of 70 % in comparison to standard protocols.

[Image quality of thin- and thick-slice MSCT reconstructions in low-contrast objects (liver lesions) with equal doses]. / Dünn- und dickschichtige MSCT-Untersuchungen bei Niedrigkontrastobjekten (Leberläsionen): Vergleich der Bildqualität bei gleicher Dosis.

PURPOSE: To evaluate the image quality of thin-section MSCT examinations of low-contrast objects such as liver lesions, and to find out whether a thin slice technique requires an increase in radiation dose. MATERIALS AND METHODS: MSCT examinations of the liver were performed in the portal venous phase on a Volume Zoom Scanner (Siemens, Erlangen) with a collimation of 4 x 2.5 mm, 120 kV, 125 mAs and table speed of 12.5 mm. Forty small hypodense liver lesions with a mean size of 3 mm (1 to 8 mm) were reconstructed using a slice thickness of 3, 5, 7, and 10 mm. All lesions were evaluated for each slice thickness by 3 independent radiologists using a 5-point scale (excellent to poor) for the following 7 criteria: contrast, demarcation and contour of the lesion, image noise and quality, demarcation of liver veins and liver contour. The mean values of the reviewers' scores were calculated. The correlation of the three observers in the evaluation of the criteria was determined using the Kendall's Tau coefficient. RESULTS: The 3-mm thin sections were excellent in the evaluation of lesion detection, lesion contrast and lesion contour with a mean score of 1.4 compared to 4.1 for 10-mm sections. Concerning the criteria image quality and liver contour, thin sections achieved the best results in our series. Image noise was pronounced in thin sections but did not affect negatively the image analysis. Interrater agreement was 0.53 for the criterium image noise. CONCLUSION: Thin sections in MSCT examinations of low contrast objects such as liver lesions do not require an increase in radiation dose because the increase in image noise is compensated by improved lesion contrast.

[Diagnostic imaging in polytrauma: comparison of radiation exposure from whole-body MSCT and conventional radiography with organ-specific CT]. / Radiologische Bildgebung beim Polytrauma: Dosisvergleich von Ganzkörper-MSCT und konventionellem Röntgen mit organspezifischer CT.

PURPOSE: To compare the radiation dose of whole-body multislice CT (MSCT) and conventional radiography with organ-specific CT in polytrauma. MATERIALS AND METHODS: The whole-body MSCT encompassing brain, neck and midface, chest, abdomen and pelvis was performed on a Somatom Volume Zoom (Siemens). Conventional radiography consisted of chest and cervical, thoracic and lumbar spine in two views as well as pelvis (Polymat, Siemens). Three combinations of organ specific CT were chosen: CT examination of (1) head and cervical spine, (2) head, cervical spine and chest, (3) head, cervical spine and abdomen. The effective doses of whole-body MSCT and conventional radiography with organ-specific CT were calculated. RESULTS: Effective doses were 20 mSv for whole-body MSCT, 2 mSv for conventional x-ray, and 5 mSv for combination (1), 8 mSv for combination (2) and (3) 16 mSv for combination (3) of the organ-specific CT. The ratio of radiation dose between whole-body MSCT and radiography was 10 : 1. This ratio was reduced to 3 : 1, 2 : 1 and 1 : 1 when a combination of radiography and CT was performed. CONCLUSIONS: Whole-body MSCT in polytrauma compared to conventional radiography with organ-specific CT induces a threefold increased dose in unfavorable situations and no increased dose in favorable situations. Nevertheless, routine use of whole-body MSCT should be critically evaluated and should be adapted to the clinical benefit.

DIRS retroelements in arthropods: identification of the recently active TcDirs1 element in the red flour beetle Tribolium castaneum.

Members of the DIRS family of retrotransposons differ from most other known retrotransposons in that they encode a tyrosine recombinase (YR), a type of enzyme frequently involved in site-specific recombination. This enzyme is believed to insert the extrachromosomal DNA intermediate of DIRS element retrotransposition into the host genome. DIRS elements have been found in plants, a slime mold, fungi, and a variety of animals including vertebrates, echinoderms and nematodes. They have a somewhat patchy distribution, however, apparently being absent from a number of model organisms such as Saccharomyces cerevisiae, Arabidopsis thaliana and Drosophila melanogaster. In this report we describe the first DIRS retroelement to be identified in an arthropod. This element, TcDirs1, was found in the red flour beetle Tribolium castaneum (Coleoptera). It is generally similar in sequence and structure to several previously described members of the DIRS group: it is bordered by inverted terminal repeats and it has a similar set of protein-coding domains (Gag, reverse transcriptase/ribonuclease H, and the YR), although these are arranged in a novel fashion. TcDirs1 elements exhibit several features indicative of recent activity, such as intact coding regions, a high level of sequence similarity between distinct elements and polymorphic insertion sites. Given their presence in an experimentally tractable host, these potentially active elements might serve as useful models for the study of DIRS element retrotransposition. An element closely related to TcDirs1 was also detected in sequences from a second arthropod, the honey bee Apis mellifera (Hymenoptera), suggesting that these retrotransposons are long-term residents of arthropod genomes.

[Multislice CT of the pelvis: dose reduction with regard to image quality]. / Mehrzeilen-Spiral CT (MSCT) des Beckenskelettes: Dosisoptimierung unter Berücksichtigung der Bildqualität.

PURPOSE: To optimize the examination protocols of multislice CT (MSCT) of the pelvis for dose reduction with regard to image quality. MATERIALS AND METHODS: MSCT of the pelvis was performed on 5 cadaver specimens with stepwise reduction of the tube current at 140 kV (250, 200, 150, 100, 75, 50, 35, 25 mAs) and 120 kV (50, 27 mAs). The remaining scan parameters like collimation (4 x 1 mm) and table speed of 4 mm per rotation remained unchanged. Axial sections and coronal reconstructions were used to evaluate cortex, trabecular structures, subjective image quality, image noise and detail detectability (pelvis and SI joint), with evaluation performed independently by four blinded experienced radiologists on a 5-point scale. Kappa coefficient, accuracy of the observers to sort the films with regard to dose reduction and mean scores of image evaluation were determined for statistical analysis. RESULTS: The deterioration of the image quality was most pronounced for all criteria between 75 mAs and 50 mAs at 140 kV. Good results with adequate image quality were obtained for detail detectability at 50 mAs and 140 kV (effective dose [E]: 3.3 mSv) and for the remaining four criteria at 75 mAs (E: 4.9 mSv). There was a moderate agreement between the four observers (kappa coefficient: 0.27). All observers were excellent in arranging the images according to the increasing dose reduction. CONCLUSIONS: Image quality of MSCT of the pelvis appears to be acceptable at 75 mAs and 140 kV with the dose reduced to 46% of the average value of the nation-wide survey of the German Roentgen Society in 1999 for this type of examination.

piggyBac-mediated germline transformation in the beetle Tribolium castaneum.

The lepidopteran transposable element piggyBac can mediate germline insertions in at least four insect orders. It therefore shows promise as a broad-spectrum transformation vector, but applications such as enhancer trapping and transposon-tag mutagenesis are still lacking. We created, cloned, sequenced and genetically mapped a set of piggyBac insertions in the red flour beetle, Tribolium castaneum. Transpositions were precise, and specifically targeted the canonical TTAA recognition sequence. We detected several novel reporter-expression domains, indicating that piggyBac could be used to identify enhancer regions. We also demonstrated that a primary insertion of a non-autonomous element can be efficiently remobilized to non-homologous chromosomes by injection of an immobile helper element into embryos harbouring the primary insertion. These developments suggest potential for more sophisticated methods of piggyBac-mediated genome manipulation.

[Quality rating of MR-cholangiopancreatography with oral application of iron oxide particles]. / Wertigkeit der MR-Cholangiopankreatikographie nach Gabe eines oralen eisenhaltigen Kontrastmittels.

PURPOSE: To compare image quality in magnetic resonance cholangiopancreatography (MRCP) performed with and without oral application of Lösferron (ferrous gluconate, Lilly Pharma, Hamburg). MATERIALS AND METHODS: A prospective study compares MRCPs performed on 52 patients with a 1.5 T clinical whole body scanner using a standard body coil. After randomization, patients ingested either 0.5 l of Lösferron (n = 27, group 1) or no oral contrast agent (n = 25, group 2) prior to the examination. 7 RARE (40 to 20 degrees) sequences were obtained, followed by selected 3 mm HASTE (T 2 -weighted with fat suppression) sequences. After blinding, image quality was rated by two radiologists using a scale of 1 (not discernible) to 5 (very well discernible). The following sections of the biliary ductal system were evaluated: left and right hepatic duct, extrahepatic bile duct and intrapancreatic bile duct. The pancreatic duct was evaluated by its location: head, body and tail of the pancreas. A Wilcoxon-Mann-Whitney test was used to determine significant differences (p < 0.05) between sampled ductal segments. Correction for multiple testing was applied. RESULTS: The oral application of Lösferron was well tolerated by all patients, and all sequences could be acquired and evaluated in all 52 patients. For the different sections of the biliary system, the mean ratings with and without Lösferron were, respectively, 3.28 and 3.36 for the left hepatic duct, 3.26 and 3.33 for the right hepatic duct, 3.46 and 4.0 for the extrahepatic bile duct, and 2.8 and 3.48 for the intrapancreatic bile duct. The corresponding ratings for the pancreatic duct were 2.8 and 3.24 for the pancreatic head, 2.84 and 3.38 for the pancreatic body, and 2.68 and 3.22 for the pancreatic tail. The differences with and without contrast agent were not statistically significant. Interobserver variability was between 0.37 for the pancreatic duct in the tail of the pancreas and 0.66 for the right hepatic duct. CONCLUSION: Despite the trend toward a better rating of the image quality for all sections of the pancreaticobiliary ductal system with Lösferron, a significant difference was not found in any ductal section after correction for multiple testing. Thus, we believe that the ingestion of Lösferron is not absolutely required prior MRCP.

Transgene expression from the Tribolium castaneum Polyubiquitin promoter.

The highly conserved Ubiquitin proteins are expressed from genes with strong, constitutively active promoters in many species, making these promoters attractive candidates for use in driving transgene expression. Here we report the cloning and characterization of the Tribolium castaneum Polyubiquitin (TcPUb) gene. We placed the TcPUb promoter upstream of the coding region of the T. castaneum eye-colour gene Tc vermilion (Tcv) and injected this construct into embryos from a Tcv-deficient strain. Transient expression of Tcv during embryogenesis resulted in complete rescue of the larval mutant phenotype. We then incorporated the TcPUb-Tcv chimera into a piggyBac donor. Resulting germline transformants were easily recognized by rescue of eye pigmentation, illustrating the potential of the TcPUb promoter for use in driving transgene expression.

Pressure evolution of the high-frequency sound velocity in liquid water.

The high-frequency sound velocity v( infinity ) of liquid water has been determined to densities of 1.37 g/cm(3) by inelastic x-ray scattering. In comparison to the hydrodynamic sound velocity v(0), the increase of v( infinity ) with density is substantially less pronounced, indicating that, at high density, the hydrogen-bond network is decreasingly relevant to the physical properties of liquid water. Furthermore, we observe an anomaly in v( infinity ) at densities around 1.12 g/cm(3), contrasting the smooth density evolution of v(0).

MR elastography of the breast:preliminary clinical results.

PURPOSE: Imaging of breast tumors and various breast tissues using magnetic resonance (MR) elastography (MRE) to explore the potential of elasticity as a new parameter for the diagnosis of breast lesions. METHODS: Low-frequency mechanical waves are transmitted into breast tissue by means of an oscillator. The local characteristics of the mechanical wave are determined by the underlying elastic properties of the tissue. Theses waves can be displayed by means of a motion-sensitive spin-echo MR sequence within the phase of the MR image. Elasticity reconstruction is performed on the basis of 8 "snapshots" of each wave within the three spatial directions. We performed in-vivo measurements in 15 female patients with malignant tumors of the breast, 5 patients with benign breast tumors, and 15 healthy volunteers. RESULTS: Malignant invasive breast tumors documented the highest values of elasticity with a median of 15.9 kPa and a wide range of stiffnesses between 8 and 28 kPa. In contrast, benign breast lesions represented low values of elasticity, which were significantly different from malignant breast tumors (median elasticity: 7.0 kPa; p = 0.0012). This was comparable to the stiffest tissue areas in healthy volunteers (median elasticity 7.0 kPa), whereas breast parenchyma (median: 2.5 kPa) and fatty breast tissue (median: 1.7 kPa) showed the lowest values of elasticity. Two invasive ductal carcinomas had elasticity values of 8 kPa and two stiff parenchyma areas in healthy volunteers had elasticities of 13 and 15 kPa. These lesions could not be differentiated by their elasticity. CONCLUSION: We conclude that MRE is a promising new imaging modality with the capability to assess the viscoelastic properties of breast tumors and the surrounding tissues. However, from our preliminary results in a small number of patients it is obvious that there is an overlap in the elasticity ranges of soft malignant tumors and stiff benign lesions.

Computer navigation of parapedicular screw fixation in the thoracic spine: a cadaver study.

STUDY DESIGN: An in vitro study to investigate the advantages of computer assistance for the purpose of parapedicular screw fixation in the upper and middle thoracic spine. OBJECTIVES: To evaluate the feasibility and application accuracy of parapedicuar screw insertion with the assistance of an optoelectronic navigation system. SUMMARY OF BACKGROUND DATA: Because of anatomic limitations, thoracic pedicle screw insertion in the upper and middle thoracic spine remains a matter of controversy. The technique of parapedicular screw insertion has been described as an alternative, although the exact screw position is difficult to control. With the assistance of computer navigation for the screw placement, it might become possible to overcome these challenges. METHODS: Four human specimens were harvested for this study; 6-mm screws were inserted from T2 to T8 with the assistance of a CT-based optoelectronic navigation system. During surgery virtual images of the screw position were documented and compared with postoperative contact radiographs to determine the application accuracy. The following measurements were obtained: axial and sagittal screw angles as well as the screw distances to the anterior vertebral cortex and the medial pedicle wall. RESULTS: All 54 screws were inserted in a parapedicular technique without violation of the medial pedicle wall or the anterior or lateral vertebral cortex. The mean +/- standard deviation difference between the virtual images and the radiographs was 1.0 +/- 0.94 mm for the distance to the medial pedicle wall and 1.9 +/- 1.44 mm for the distance to the anterior cortex. The angular measurements showed a difference of 1.6 +/- 1.1 degrees for the transverse screw angle and 2.1 +/- 1.6 degrees for the sagittal screw orientation. CONCLUSION: With the assistance of computer navigation it is possible to achieve a safe and reliable parapedicular screw insertion in the upper and middle thoracic spine in vitro. The application accuracy varies for the linear and angular measurements and is higher in the axial than in the sagittal plane. It is important for the surgeon to understand these limitations when using computer navigation in spinal surgery.

In vitro function of islets of Langerhans encapsulated with a membrane of porcine chondrocytes for immunoisolation.

BACKGROUND/AIMS: Widespread clinical application of islet transplantation remains restricted, because of insufficient methods to prevent rejection and autoimmune destruction of islet grafts. In this study we demonstrate long-term function of islets of Langerhans within a capsule of porcine chondrocytes which may serve as an immunoisolation barrier utilizing the immunoprivileged properties of the chondrocyte matrix. METHODS: Islets of Langerhans were isolated from Lewis rats, seeded on biodegradable polyglycolic acid polymer, and encapsulated with a monolayer of porcine chondrocytes. The encapsulated constructs and controls were kept in culture for 5 weeks. One group was exposed to a glucose challenge every 5th day. The insulin concentration of the culture medium was measured. Histological and insulin-immunohistochemical studies were performed. RESULTS: Hematoxylin and eosin histology demonstrated viability of the islets of Langerhans. The intact morphology was demonstrated by Heidenhain staining. Toluidine blue showed viability of surrounding chondrocyte layers. Immunohistochemistry was positive for insulin within the beta cells of the islets. Both encapsulated constructs and nonencapsulated controls showed increasing insulin levels after glucose challenge. CONCLUSIONS: We can tissue engineer a chondrocyte encapsulation membrane which permits diffusion of glucose and insulin. Islets of Langerhans survive within the chondrocyte capsule, and the glucose/insulin feedback mechanism remains intact.