Protective role of vascular endothelial growth factor in endotoxin-induced acute lung injury in mice

<p>Abstract</p> <p>Background</p> <p>Vascular endothelial growth factor (VEGF), a substance that stimulates new blood vessel formation, is an important survival factor for endothelial cells. Although overexpressed VEGF in the lung induces pulmonary edema with increased lung vascular permeability, the role of VEGF in the development of acute lung injury remains to be determined.</p> <p>Methods</p> <p>To evaluate the role of VEGF in the pathogenesis of acute lung injury, we first evaluated the effects of exogenous VEGF and VEGF blockade using monoclonal antibody on LPS-induced lung injury in mice. Using the lung specimens, we performed TUNEL staining to detect apoptotic cells and immunostaining to evaluate the expression of apoptosis-associated molecules, including caspase-3, Bax, apoptosis inducing factor (AIF), and cytochrome C. As a parameter of endothelial permeability, we measured the albumin transferred across human pulmonary artery endothelial cell (HPAEC) monolayers cultured on porous filters with various concentrations of VEGF. The effect of VEGF on apoptosis HPAECs was also examined by TUNEL staining and active caspase-3 immunoassay.</p> <p>Results</p> <p>Exogenous VEGF significantly decreased LPS-induced extravascular albumin leakage and edema formation. Treatment with anti-VEGF antibody significantly enhanced lung edema formation and neutrophil emigration after intratracheal LPS administration, whereas extravascular albumin leakage was not significantly changed by VEGF blockade. In lung pathology, pretreatment with VEGF significantly decreased the numbers of TUNEL positive cells and those with positive immunostaining of the pro-apoptotic molecules examined. VEGF attenuated the increases in the permeability of the HPAEC monolayer and the apoptosis of HPAECs induced by TNF-α and LPS. In addition, VEGF significantly reduced the levels of TNF-α- and LPS-induced active caspase-3 in HPAEC lysates.</p> <p>Conclusion</p> <p>These results suggest that VEGF suppresses the apoptosis induced by inflammatory stimuli and functions as a protective factor against acute lung injury.</p

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Role of toll-like receptor 4 in acute neutrophilic lung inflammation induced by intratracheal bacterial products in mice

Wakako Yamada1, Sadatomo Tasaka1, Hidefumi Koh1, Mie Shimizu1, Yuko Ogawa1, Naoki Hasegawa1, Taku Miyasho2, Kazuhiro Yamaguchi1, Akitoshi Ishizaka11Division of Pulmonary Medicine, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan; 2Laboratory of Veterinary Biochemistry, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, JapanBackground: Toll-like receptors (TLRs) represent a conserved family of innate immune recognition receptors. Among TLRs, TLR4 is important for the recognition of Gram-negative bacteria, whereas TLR2 recognizes cell wall constituents of Gram-positive microorganisms, such as peptidoglycan (PGN).Methods: To evaluate the role of TLR4 in the pathogenesis of acute lung injury induced by Escherichia coli endotoxin (lipopolysaccharide; LPS) or PGN, we compared inflammatory cell accumulation in bronchoalveolar lavage (BAL) fluid and lung pathology between C3H/HeJ (TLR4 mutant) and wild-type C3H/HeN mice. The levels of proinflammatory cytokines and chemokines in plasma and BAL fluid and nuclear factor-&amp;kappa;B (NF-&amp;kappa;B) translocation in the lung were also evaluated.Results: In C3H/HeJ mice, LPS-induced neutrophil emigration was significantly decreased compared with C3H/HeN mice, whereas PGN-induced neutrophil emigration did not differ. Differential cell count in BAL fluid revealed comparable neutrophil recruitment in the alveolar space. In TLR4 mutant mice, LPS-induced upregulation of tumor necrosis factor-alpha (TNF-&amp;alpha;), KC, and CXCL10 in plasma and BAL fluid was attenuate, which was not different after PGN. NF-&amp;kappa;B translocation in the lung was significantly decreased in C3H/HeJ compared with C3H/HeN mice, whereas PGN-induced NF-&amp;kappa;B translocation was not different.Conclusion: These results suggest that TLR4 mediates inflammatory cascade induced by Gram-negative bacteria that is locally administered.Keywords: rodent, TLR4, endotoxin, neutrophils, NF-&amp;kappa;

THE EFFECTS OF TRIAZOLAM AND RILMAZAFONE ON THE PHYSICAL AND COGNITIVE FUNCTIONS IN HEALTHY ELDERLY PERSONS

The purpose of this study is to investigate whether often used hypnotics are safe or not to elderlypeople by evaluating the early morning effects and residual effects on the physical and cognitivefunctions after hypnotic administration.Fourteen healthy elders (3 male, 11 female ; mean age 64 years) took part in the study. Triazolam0.125 mg, rilmazafone hydrochloride 1 mg or placebo was orally given to each subjectbefore going to bed (11 pm). Objective evaluations including 1. Total Sway Path test (openedand closed eye)　2. Functional Reach test　3. Timed Up and Go test　4. Simply DiscriminatoryReaction test　5. Critical Flicker Fusion test　6. Short-Term Memory test were done at 10 pm(an hour before subjects went to bed), 4 am, 6 am, 10 am and 2 pm.Among all evaluations for the physical and cognitive functions, main effects between medicineswere only seen in the Total Sway Path test, which is the indicator for static balance ability. Wheneyes were opened, trembling was significantly small in the rilmazafon group compared to the placebogroup (p=0.04).In this study, the hypnotics did not affect physical and cognitive functions for healthy elders. Furtherstudies using an interdisciplinary approach with different professionals such nurses, physicaltherapists, pharmacists and physicians are needed for evaluating the safety use of hypnotics in theelderly patients with insomnia