.-In the lungs, high-pressure mechanical ventilation induces an inflammatory response similar to that observed in acute respiratory distress syndrome. To further characterize these responses and to compare them with classical inflammatory pathways, we performed gene expression profiling analysis of 20,000 mouse genes in isolated blood-free (to exclude genes from sequestered leukocytes) perfused mouse lungs exposed to lowpressure ventilation (10 cmH2O), high-pressure ventilation (25 cmH2O, overventilation), and LPS treatment. A large number of inflammatory and apoptotic genes were increased by both overventilation and LPS. However, certain growth factor-related genes, as well as genes related to development, cellular communication, and the cytoskeleton, were only regulated by overventilation. We validated and confirmed increased mRNA expression pattern of five genes (amphiregulin, gravin, Nur77, Cyr61, interleukin-11) by real-time PCR; furthermore, we confirmed increased protein expression of amphiregulin by immunohistochemistry and immunoblotting assays. These genes represent novel candidate genes in ventilator-induced lung injury. overventilation; acute respiratory distress syndrome; gene expression analysis DESPITE INTENSIVE INVESTIGATIONS, the mortality of acute respiratory distress syndrome (ARDS) remains high (1a). Patients at risk for ARDS as well as ARDS patients themselves (13, 20) are prone to ventilator-induced lung injury (VILI) and benefit from low tidal volume ventilation (1a). Clinical and animal studies suggest that the release of proinflammatory cytokines and other factors during alveolar overdistension contributes to lung injury in VILI (1a,9,12,17). One important insight in this area is that ventilation with high distending pressures (overventilation) may activate, if by different signaling mechanisms, canonical inflammation pathways that also become activated by well-known proinflammatory stimuli such as bacterial endotoxin (31). For instance, both overventilation and LPS activate nuclear factor-B (NF-B) (17, 32) and mitogen-activated protein kinase (MAPK) (33), which subsequently activate chemokines, cytokines, and adhesion molecules (32). One important objective is to identify genes that are specifically activated by overventilation but not by endotoxin. Such genes might provide specific targets to reduce the side effects of mechanical ventilation without interfering with the innate immune system. Microarray technology allows us to study gene expression of a significant fraction of the genome and has been used to identify candidate genes critical to a variety of lung diseases including chronic obstructive pulmonary disease, idiopathic pulmonary fibrosis, lung cancer, pulmonary hypertension, and lung transplantation (2,8,24,36,37). However, application of this approach to ventilator-induced gene expression in vivo is complicated by at least two factors: 1) in intact noninjured animals, alveolar overdistension is limited by both the chest wall and adjacent alveoli, and 2) infiltrating inflam...

Cupin-type phosphoglucose isomerases (cPGIs) were identified in some archaeal and bacterial genomes and the respective coding function of cpgi's from the euryarchaeota Archaeoglobus fulgidus and Methanosarcina mazei, as well as the bacteria Salmonella enterica serovar Typhimurium and Ensifer meliloti, was proven by functional overexpression. These cPGIs and the cPGIs from Pyrococcus and Thermococcus spp. represent the cPGI family and were compared with respect to kinetic, inhibitory, thermophilic, and metal-binding properties. cPGIs showed a high specificity for the substrates fructose-6-phosphate and glucose-6-phosphate and were inhibited by millimolar concentrations of sorbitol-6-phosphate, erythrose-4-phosphate, and 6-phosphogluconate. Treatment of cPGIs with EDTA resulted in a complete loss of catalytic activity, which could be regained by the addition of some divalent cations, most effectively by Fe 2؉ and Ni 2؉ , indicating a metal dependence of cPGI activity. The motifs TX 3 PX 3 GXEX 3 TXGHXHX 6-11 EXY and PPX 3 HX 3 N were deduced as the two signature patterns of the novel cPGI family. Phylogenetic analysis suggests lateral gene transfer for the bacterial cPGIs from euryarchaeota.Phosphoglucose isomerase (PGI; EC 5.3.1.9) catalyzes the reversible isomerization of glucose-6-phosphate to fructose-6-phosphate. PGI plays a central role in sugar metabolism of eukarya, bacteria, and archaea, both in glycolysis via the Embden-Meyerhof pathway in eukarya and bacteria and in its modified versions found in archaea. PGI is also involved in gluconeogenesis, where the enzyme operates in the reverse direction (see references 22, 26, and 40). PGIs have evolved convergently. Most PGIs belong to the PGI superfamily, which can be divided into the PGI family and the recently identified bifunctional phosphoglucose/phosphomannose isomerase (PGI/PMI) family (22). PGIs from the PGI superfamily, often referred to as conventional PGIs, are found in all domains of life and are well studied. Crystal structures have been determined for the eukaryotic PGIs from pigs, rabbits, humans and from the bacterium Bacillus stearothermophilus, and conserved amino acids proposed to be involved in substrate binding and/ or catalysis have been identified (3, 6, 10-13, 28, 29, 42). Bifunctional PGI/PMIs, which have been characterized as a novel family within the PGI superfamily, were predominantly found in the crenarchaeotal branch of the archaea (26).Recently, a novel type of PGI has been identified and characterized from the hyperthermophilic euryarchaeon Pyrococcus furiosus (22, 52) and later from the closely related Thermococcus litoralis (30). These PGIs belong to the cupin superfamily and thus represent a convergent line of PGI evolution. The cupin superfamily is present in all three domains of life-eukarya, bacteria, and archaea-and comprises a group of functionally diverse proteins that contain a central domain composed of ␤-strands forming a small ␤-barrel called "cupin."Proteins from the cupin superfamily range, for example, from manno...