Targeted disruption of fibulin-4 abolishes elastogenesis and causes perinatal lethality in mice

Elastic fibers provide tissues with elasticity which is critical to the function of arteries, lungs, skin, and other dynamic organs. Loss of elasticity is a major contributing factor in aging and diseases. However, the mechanism of elastic fiber development and assembly is poorly understood. Here, we show that lack of fibulin-4, an extracellular matrix molecule, abolishes elastogenesis. fibulin-4(−/−) mice generated by gene targeting exhibited severe lung and vascular defects including emphysema, artery tortuosity, irregularity, aneurysm, rupture, and resulting hemorrhages. All the homozygous mice died perinatally. The earliest abnormality noted was a uniformly narrowing of the descending aorta in fibulin-4(−/−) embryos at embryonic day 12.5 (E12.5). Aorta tortuosity and irregularity became noticeable at E15.5. Histological analysis demonstrated that fibulin-4(−/−) mice do not develop intact elastic fibers but contain irregular elastin aggregates. Electron microscopy revealed that the elastin aggregates are highly unusual in that they contain evenly distributed rod-like filaments, in contrast to the amorphous appearance of normal elastic fibers. Desmosine analysis indicated that elastin cross-links in fibulin-4(−/−) tissues were largely diminished. However, expression of tropoelastin or lysyl oxidase mRNA was unaffected in fibulin-4(−/−) mice. In addition, fibulin-4 strongly interacts with tropoelastin and colocalizes with elastic fibers in culture. These results demonstrate that fibulin-4 plays an irreplaceable role in elastogenesis

Preliminary evaluation of a robotic apparatus for the analysis of passive glenohumeral joint kinematics

Background: The shoulder has the greatest range of motion of any joint in the human body. This is due, in part, to the complex interplay between the glenohumeral (GH) joint and the scapulothoracic (ST) articulation. Currently, our ability to study shoulder kinematics is limited, because existing models isolate the GH joint and rely on manual manipulation to create motion, and have low reproducibility. Similarly, most established techniques track shoulder motion discontinuously with limited accuracy. Methods: To overcome these problems, we have designed a novel system in which the shoulder girdle is studied intact, incorporating both GH and ST motions. In this system, highly reproducible trajectories are created using a robotic actuator to control the intact shoulder girdle. High-speed cameras are employed to track retroreflective bone markers continuously. Results: We evaluated this automated system’s capacity to reproducibly capture GH translation in intact and pathologic shoulder conditions. A pair of shoulders (left and right) were tested during forward elevation at baseline, with a winged scapula, and after creation of a full thickness supraspinatus tear. Discussion The system detected differences in GH translations as small as 0.5 mm between different conditions. For each, three consecutive trials were performed and demonstrated high reproducibility and high precision

Joint effect of phosphorus limitation and temperature on alkaline phosphatase activity and somatic growth in Daphnia magna

Alkaline phosphatase (AP) is a potential biomarker for phosphorus (P) limitation in zooplankton. However, knowledge about regulation of AP in this group is limited. In a laboratory acclimation experiment, we investigated changes in body AP concentration for Daphnia magna kept for 6 days at 10, 15, 20 and 25°C and fed algae with 10 different molar C:P ratios (95–660). In the same experiment, we also assessed somatic growth of the animals since phosphorus acquisition is linked to growth processes. Overall, non-linear but significant relationships of AP activity with C:P ratio were observed, but there was a stronger impact of temperature on AP activity than of P limitation. Animals from the lowest temperature treatment had higher normalized AP activity, which suggests the operation of biochemical temperature compensation mechanisms. Body AP activity increased by a factor of 1.67 for every 10°C decrease in temperature. These results demonstrate that temperature strongly influences AP expression. Therefore, using AP as a P limitation marker in zooplankton needs to consider possible confounding effects of temperature. Both temperature and diet affected somatic growth. The temperature effect on somatic growth, expressed as the Q10 value, responded non-linearly with C:P, with Q10 ranging between 1.9 for lowest food C:P ratio and 1.4 for the most P-deficient food. The significant interaction between those two variables highlights the importance of studying temperature-dependent changes of growth responses to food quality

The Complete Genome Sequence of the Pathogenic Intestinal Spirochete Brachyspira pilosicoli and Comparison with Other Brachyspira Genomes

Background: The anaerobic spirochete Brachyspira pilosicoli colonizes the large intestine of various species of birds and mammals, including humans. It causes ''intestinal spirochetosis'', a condition characterized by mild colitis, diarrhea and reduced growth. This study aimed to sequence and analyse the bacterial genome to investigate the genetic basis of its specialized ecology and virulence. Methodology/Principal Findings: The genome of B. pilosicoli 95/1000 was sequenced, assembled and compared with that of the pathogenic Brachyspira hyodysenteriae and a near-complete sequence of Brachyspira murdochii. The B. pilosicoli genome was circular, composed of 2,586,443 bp with a 27.9 mol% G+C content, and encoded 2,338 genes. The three Brachyspira species shared 1,087 genes and showed evidence of extensive genome rearrangements. Despite minor differences in predicted protein functional groups, the species had many similar features including core metabolic pathways. Genes distinguishing B. pilosicoli from B. hyodysenteriae included those for a previously undescribed bacteriophage that may be useful for genetic manipulation, for a glycine reductase complex allowing use of glycine whilst protecting from oxidative stress, and for aconitase and related enzymes in the incomplete TCA cycle, allowing glutamate synthesis and function of the cycle during oxidative stress. B. pilosicoli had substantially fewer methyl-accepting chemotaxis genes than B. hyodysenteriae and hence these species are likely to have different chemotactic responses that may help to explain their different host range and colonization sites. B. pilosicoli lacked the gene for a new putative hemolysin identified in B. hyodysenteriae WA1. Both B. pilosicoli and B. murdochii lacked the rfbBADC gene cluster found on the B. hyodysenteriae plasmid, and hence were predicted to have different lipooligosaccharide structures. Overall, B. pilosicoli 95/1000 had a variety of genes potentially contributing to virulence. Conclusions/Significance: The availability of the complete genome sequence of B. pilosicoli 95/1000 will facilitate functional genomics studies aimed at elucidating host-pathogen interactions and virulence

Persistent Gastric Colonization with Burkholderia pseudomallei and Dissemination from the Gastrointestinal Tract following Mucosal Inoculation of Mice

Melioidosis is a disease of humans caused by opportunistic infection with the soil and water bacterium Burkholderia pseudomallei. Melioidosis can manifest as an acute, overwhelming infection or as a chronic, recurrent infection. At present, it is not clear where B. pseudomallei resides in the mammalian host during the chronic, recurrent phase of infection. To address this question, we developed a mouse low-dose mucosal challenge model of chronic B. pseudomallei infection and investigated sites of bacterial persistence over 60 days. Sensitive culture techniques and selective media were used to quantitate bacterial burden in major organs, including the gastrointestinal (GI) tract. We found that the GI tract was the primary site of bacterial persistence during the chronic infection phase, and was the only site from which the organism could be consistently cultured during a 60-day infection period. The organism could be repeatedly recovered from all levels of the GI tract, and chronic infection was accompanied by sustained low-level fecal shedding. The stomach was identified as the primary site of GI colonization as determined by fluorescent in situ hybridization. Organisms in the stomach were associated with the gastric mucosal surface, and the propensity to colonize the gastric mucosa was observed with 4 different B. pseudomallei isolates. In contrast, B. pseudomallei organisms were present at low numbers within luminal contents in the small and large intestine and cecum relative to the stomach. Notably, inflammatory lesions were not detected in any GI tissue examined in chronically-infected mice. Only low-dose oral or intranasal inoculation led to GI colonization and development of chronic infection of the spleen and liver. Thus, we concluded that in a mouse model of melioidosis B. pseudomallei preferentially colonizes the stomach following oral inoculation, and that the chronically colonized GI tract likely serves as a reservoir for dissemination of infection to extra-intestinal sites

Optics and Quantum Electronics

Contains table of contents on Section 3 and reports on nineteen research projects.Defense Advanced Research Projects Agency Grant F49620-96-0126Joint Services Electronics Program Grant DAAH04-95-1-0038National Science Foundation Grant ECS 94-23737U.S. Air Force - Office of Scientific Research Contract F49620-95-1-0221U.S. Navy - Office of Naval Research Grant N00014-95-1-0715Defense Advanced Research Projects Agency/National Center for Integrated Photonics TechnologyMultidisciplinary Research InitiativeU.S. Air Force - Office of Scientific ResearchNational Science Foundation/MRSECU.S. Navy - Office of Naval Research (MFEL) Contract N00014-91-J-1956National Institutes of Health Grant R01-EY11289U.S. Navy - Office of Naval Research (MFEL) Contract N00014-94-0717Defense Advanced Research Projects Agency Contract N66001-96-C-863

Optics and Quantum Electronics

Contains table of contents for Section 3 and reports on eighteen research projects.Defense Advanced Research Projects Agency/MIT Lincoln Laboratory Contract MDA972-92-J-1038Joint Services Electronics Program Grant DAAH04-95-1-0038National Science Foundation Grant ECS 94-23737U.S. Air Force - Office of Scientific Research Contract F49620-95-1-0221U.S. Navy - Office of Naval Research Grant N00014-95-1-0715MIT Center for Material Science and EngineeringNational Center for Integrated Photonics Technology Contract DMR 94-00334National Center for Integrated Photonics TechnologyU.S. Navy - Office of Naval Research (MFEL) Contract N00014-94-1-0717National Institutes of Health Grant 9-R01-EY11289MIT Lincoln Laboratory Contract BX-5098Electric Power Research Institute Contract RP3170-25ENEC

Optics and Quantum Electronics

Contains table of contents for Section 3 and reports on twenty research projects.Charles S. Draper Laboratories Contract DL-H-467138Joint Services Electronics Program Contract DAAL03-92-C-0001Joint Services Electronics Program Grant DAAH04-95-1-0038U.S. Air Force - Office of Scientific Research Contract F49620-91-C-0091MIT Lincoln LaboratoryNational Science Foundation Grant ECS 90-12787Fujitsu LaboratoriesNational Center for Integrated PhotonicsHoneywell Technology CenterU.S. Navy - Office of Naval Research (MFEL) Contract N00014-94-1-0717U.S. Navy - Office of Naval Research (MFEL) Grant N00014-91-J-1956National Institutes of Health Grant NIH-5-R01-GM35459-09U.S. Air Force - Office of Scientific Research Grant F49620-93-1-0301MIT Lincoln Laboratory Contract BX-5098Electric Power Research Institute Contract RP3170-25ENEC