Previous studies have found that inappropriate elevation of matrix metalloproteinase-9 (MMP9) expression and activity is coincident with early onset of emphysema in Smad3-null mice. Herein, we further investigated the role of increased MMP9 in emphysema pathogenesis and the related molecular regulatory mechanisms of elevated MMP9 in Smad3-null lung. Genetic blockade of MMP9 in Smad3-null mice significantly attenuated emphysema pathology, but not hypoalveolarization during early postnatal lung development. Furthermore, Smad3 was found to be a transcription factor to positively regulate a protein deacetylase SIRT1 by binding to an AP-1 site of SIRT1 promoter. A synergistic regulatory effect on SIRT1 expression was also detected between Smad3 and c-Jun. Consistently, Smad3 knockout lung at P28 had reduced SIRT1 expression, which in turn resulted in increased acetylation of histone H3 at the transcription factor AP-1, NF-κB, and Pea3 binding sites of MMP9 promoter and increased acetylation of NF-κB. In addition, increased Pea3 expression and nuclear accumulation was also detected in Smad3-null lungs at P28. Consistently, bindings of acetylated NF-κB and Pea3 to the MMP9 promoter were elevated in Smad3-null lung. We thus propose that deficiency of Smad3 causes downregulation of SIRT1 and increased Pea3 expression/nuclear accumulation, respectively. Decreased SIRT1 activity resulted in increased acetylation of histone H3 and NF-κB. Subsequently, increased bindings of transcription factors including NF-κB and Pea3 to MMP9 promoter significantly upregulate MMP9 transcription, contributing to emphysema pathogenesis.

OBJECTIVE: Transferred CD4(+) CD25(+) Foxp3(+) regulatory cells (Tregs) can prevent autoimmune disease, but generally fail to ameliorate established disease. Here we demonstrate that antigen-specific Tregs induced with IL-2 and TGF-β ex-vivo (iTregs), but not equivalent expanded thymus-derived nTregs can prevent progression of established collagen-induced arthritis. METHODS: DBA/1 mice with collagen-induced arthritis were treated with iTreg or nTreg generated or expanded in vitro before or shortly after collagen II immunization, and clinical scores were determined. Inflammatory responses were determined by measuring the levels of anti-CII antibody in the serum and histological pathology in joints. The Th1/Th17-mediated autoreactive response was evaluated by determining the cytokine profile of draining lymph node cells by flow cytometry. RESULTS: Following transfer, nTregs exhibited decreased Foxp3 and Bcl-2 expression, decreased suppressive activity, and many converted to Th17 cells. By contrast, transferred iTregs were more numerous, retained Foxp3 expression and their suppressive activity in the presence of IL-6, and were resistant to Th17 conversion. Remarkably, ten days after transfer of donor iTregs shifted the predominance from Th17 to Treg cells in recipient draining LNs. CONCLUSION: These findings provide evidence that transferred TGF-β-induced iTregs are more stable and functional than nTregs in mice with established autoimmunity. Moreover, iTregs can have tolerogenic effects even in the presence of ongoing inflammation. The therapeutic potential of human iTregs in subjects with chronic, immune-mediated inflammatory diseases deserves to be investigated.

The RV144 trial demonstrated that an experimental AIDS vaccine can prevent human immunodeficiency virus-1 (HIV-1) infection in humans. Because of its limited efficacy, further understanding of the mechanism of preventive AIDS vaccines remains a priority, and non-human primate models of lentiviral infection provide an opportunity to define immunogens, vectors, and correlates of immunity. In this study, we show that prime-boost vaccination with a mis-matched HIV-1 Envelope (env) gene, derived from SIVmac239, prevents infection by the SIVsmE660 virus intra-rectally. Analysis of different gene-based prime-boost immunization regimens revealed that recombinant adenovirus 5 (rAd5) prime followed by replication-defective lymphocytic choriomeningitis virus (rLCMV) boost elicited robust CD4 and CD8 T-cell and humoral immune responses. This vaccine protected against infection after repetitive mucosal challenge with an efficacy of 82% per exposure and 62% cumulatively. No effect was seen on viremia in infected vaccinated monkeys compared to controls. Protection correlated with the presence of neutralizing antibodies to the challenge viruses tested in peripheral blood mononuclear cells. These data indicate that a vaccine expressing a mis-matched Env gene alone can prevent SIV infection in NHP and identifies an immune correlate that may guide immunogen selection and immune monitoring for clinical efficacy trials.

It is desirable to obtain the maximum assist without suction in ventricular assist devices (VADs). However, high driving power of a VAD may cause severe ventricle suction that can induce arrhythmia, hemolysis, and pump damage. In this report, an appropriate VAD driving level that maximizes the assist effect without severe systolic suction was explored. The target driving level was set at the boundary between low driving power without suction and high driving power with frequent suction. In the boundary range, intermittent mild suction may occur. Driving power was regulated by the suction occurrence. The normal-suction boundary control method was evaluated in a female goat implanted with an undulation pump ventricular assist device (UPVAD). The UPVAD was driven in a semipulsatile mode with heartbeat synchronization control. Systolic driving power was adjusted using a normal-suction boundary control method developed for this study. We confirmed that driving power could be maintained in the boundary range. Occurrences of suction were evaluated using the suction ratio. We defined this ratio as the number of suction occurrences divided by the number of heartbeats. The suction ratio decreased by 70% when the normal-suction boundary control method was used.

BACKGROUND A high level of matrix metalloproteinase-9 (MMP-9) is associated with human tumor invasion and/or metastasis. The HT1080 human fibrosarcoma cell line is highly invasive and metastatic which constitutively express MMP-9. METHODS HT1080 cells transfected with a double stranded RNA that targeted the MMP-9 mRNA and the cellular characteristics were examined before and after interference. The inhibition effects of MMP-9 interference on the tumor growth of HT1080 cells in nude mice was also tested by xenograft assay. RESULTS MMP-9 extinction in HT1080 resulted in the following:(1) inhibited cell mobility;(2) increased cell adhesion, and (3) attenuated tumor cell migration. In addition, MMP-9 knockdown concomitantly resulted in decreased levels of soluble ICAM-1, leading to an adhesion defect and tumor metastasis. Moreover, in vivo assay further demonstrated MMP-9 interference affecting the tumorigenesis of HT1080 cells in mice as follows (1) inhibition of tumor growth;(2) reduced tumor volume, and (3) prolonged survival time. CONCLUSIONS Our observations defined a novel critical role for MMP-9 in the progression of HT1080 fibrosarcoma by changing the inter-cellular adhesion molecular-1 from membrane-anchored state to a soluble one which provides a target for promising tumor therapy in clinics.

Objective: To compare same-day and next-day pain control and safety of two anesthetic techniques utilizing 4% liquid lidocaine applied with sterile cotton swabs versus 3.5% lidocaine gel for intravitreal injections. Main outcome measures were: discomfort during anesthetic preparation and needle penetration, 1 and 24 h after injection. Methods: Patients were randomized to alternate anesthetic method at two consecutive injections in one eye or in different eyes on the same day if requiring bilateral injections. Overall satisfaction, corneal staining, and subconjunctival hemorrhage (SCH) were compared. Results: Fifty patients were enrolled. Both methods resulted in similar mild discomfort during anesthetic preparation, 1 and 24 h later. The gel resulted in slightly higher discomfort during needle penetration (p = 0.026). Patients were satisfied with both techniques (p = 0.91), however, 52% patients preferred gel, 33% were indifferent, and 15% preferred cotton swabs (p = 0.002). There were significantly less corneal staining (p = 0.001) and SCH (p = 0.004) after the gel. Conclusion: Both techniques are equally effective and yield mild discomfort scores during the procedure and the next day. The gel method results in significantly less ocular surface irritation.

The outbreak of highly pathogenic avian influenza (HPAI) H5N1 disease has led to significant loss of poultry and wild life and case fatality rates in humans of 60%. Wild birds are natural hosts for all avian influenza virus subtypes and over120 bird species have been reported with evidence of H5N1 infection. Influenza A viruses possess a segmented RNA genome and are characterized by frequently occurring genetic reassortment events, which play a very important role in virus evolution and the spread of novel gene constellations in immunologically naïve human and animal populations. Phylogenetic analysis of whole genome or sub-genomic sequences is a standard means for delineating genetic variation, novel reassortment events, and surveillance to trace the global transmission pathways. In this paper, special emphasis is given to the transmission and circulation of H5N1 among wild life populations, and to the reassortment events that are associated with inter-host transmission of the H5N1 viruses when they infect different hosts, such as birds, pigs and humans. In addition, we review the inter-subtype reassortment of the viral segments encoding inner proteins between the H5N1 viruses and viruses of other subtypes, such as H9N2 and H6N1. Finally, we highlight the usefulness of genomic sequences in molecular epidemiological analysis of HPAI H5N1 and the technical limitations in existing analytical methods that hinder them from playing a greater role in virological research.

The crystallinity, textural characteristics, optical absorption properties, as well as the photocatalytic hydrogen production activities of three-dimensional interconnected mesoporous In(2)O(3)/Ta(2)O(5) composites were investigated as functions of In(2)O(3) content (x) and calcination temperature (T). The results show that the incorporation of In(2)O(3) endows intimate heterostructured junctions in the composites and significantly improves the thermal stability of mesopores. With increasing x, the as-prepared composites possess similar textural properties and continuously increased light absorbencies, but a maximum heterojunction area, and thus, the optimal value for the average hydrogen evolution rate ([Formula: see text]) as well as the special surface hydrogen evolution rate ([Formula: see text]) at x=20%. For the 20% In(2)O(3)/Ta(2)O(5) composites prepared at different T, the special surface area decreases and the pore size enlarges with increasing T from 450 to 650°C. An obvious collapse of mesopores accompanying with remarkable crystallization occurs at 750°C. The highest [Formula: see text] occurs on the sample calcined at 550°C, while the optimal [Formula: see text] appears at 750°C. This suggests that good charge carrier separation and transport properties, rapid mass transfer of reactants and gases desorption are as important as large surface area and high crystallinity for the photocatalysts.