Cross-talk between human mast cells and bronchial epithelial cells in plasminogen activator inhibitor-1 production via transforming growth factor-ß1.

Previous reports suggest that plasminogen activator inhibitor-1 (PAI-1) promotes airway remodeling and that human and mouse mast cells (MCs) are an important source of PAI-1. In the present study we investigated MC-epithelial cell (EC) interactions in the production of PAI-1. We stimulated the human MC line LAD2 with IgE-receptor cross-linking and collected the supernatants. We incubated the human bronchial EC line BEAS-2B with the LAD2 supernatants and measured the level of PAI-1. When the supernatants from IgE-stimulated LAD2 were added to BEAS-2B, there was a significant enhancement of PAI-1 production by BEAS-2B. When we treated the MC supernatants with a transforming growth factor (TGF)-ß1 neutralizing antibody, the MC-derived induction of PAI-1 from BEAS-2B was completely abrogated. Although TGF-ß1 mRNA was constitutively expressed in resting LAD2, it was not highly induced by IgE-mediated stimulation. Nonetheless, active TGF-ß1 protein was significantly increased in LAD2 after IgE-mediated stimulation. Active TGF-ß1 produced by primary cultured human MCs was significantly reduced in the presence of a chymase inhibitor, suggesting a role of MC chymase as an activator of latent TGF-ß1. This study indicates that stimulation of human MCs by IgE receptor cross-linking triggers activation of TGF-ß1, at least in part via chymase, which in turn induces the production of PAI-1 by bronchial ECs. Our data suggest that human MCs may play an important role in airway remodeling in asthma as a direct source of PAI-1 and by activating bronchial ECs to produce further PAI-1 via a TGF-ß1-mediated activation pathway.

Age-related increased prevalence of asthma and nasal polyps in chronic rhinosinusitis and its association with altered IL-6 trans-signaling.

We report that S100 proteins were reduced in patients with chronic rhinosinusitis (CRS). S100A8/9, which is important in epithelial barrier function, was particularly decreased in elderly patients with CRS. Epithelial expression of S100A8/9 is partly regulated by the IL-6 trans-signaling pathway. The goal of this study was to investigate whether or not age-related reduction of S100A8/9 in CRS is associated with blunting of IL-6 trans-signaling. The levels of IL-6, soluble IL-6 receptor (sIL-6R), soluble gp130 (sgp130), and S100A8/9 from control subjects (n = 10), and patients with CRS without nasal polyps (n = 13) and those with CRS with nasal polyps (CRSwNP) (n = 14), were measured by ELISA. Age-related differences in the level of each protein were investigated. Normal human bronchial epithelial cells were cultured in air-liquid interface and stimulated with IL-6/sIL-6R and tumor necrosis factor (TNF)-α with or without the addition of sgp130, a natural inhibitor of IL-6 trans-signaling. There was a significant age-related decline in S100A8/9 and an increase in sgp130 in nasal tissue samples from patients with CRSwNP, although there was no age-related difference in IL-6/sIL-6R production. Additionally, expression of the S100A8/9 gene and protein was increased significantly by IL-6/sIL-6R plus TNF-α in normal human bronchial epithelial cells. This increase was blocked by sgp130. These results suggest that increased sgp130 in older patients may inhibit IL-6 trans-signaling, impair barrier function, and decrease S1008/9 production in elderly patients with CRSwNP. Restoration of barrier function by targeting sgp130 may be a novel treatment strategy.

Temperature and magnetic field effects on electron transport through DNA molecules in a two-dimensional four-channel system.

We utilize a two-dimensional four-channel DNA model, with a tight-binding (TB) Hamiltonian, and investigate the temperature and the magnetic field dependence of the transport behavior of a short DNA molecule. Random variation of the hopping integrals due to the thermal structural disorder, which partially destroy phase coherence of electrons and reduce quantum interference, leads to a reduction of the localization length and causes suppressed overall transmission. We also incorporate a variation of magnetic field flux density into the hopping integrals as a phase factor and observe Aharonov-Bohm (AB) oscillations in the transmission. It is shown that for non-zero magnetic flux, the transmission zero leaves the real-energy axis and moves up into the complex-energy plane. We also point out that the hydrogen bonds between the base pair with flux variations play a role to determine the periodicity of AB oscillations in the transmission.

A plasminogen activator inhibitor-1 inhibitor reduces airway remodeling in a murine model of chronic asthma.

We previously reported that plasminogen activator inhibitor (PAI)-1 deficiency prevents collagen deposition in the airways of ovalbumin (OVA)-challenged mice. In this study, we explored the therapeutic utility of blocking PAI-1 in preventing airway remodeling, using a specific PAI-1 inhibitor, tiplaxtinin. C57BL/6J mice were immunized with intraperitoneal injections of OVA on Days 0, 3, and 6. Starting on Day 11, mice were challenged with phosphate-buffered saline or OVA by nebulization three times per week for 4 weeks. Tiplaxtinin was mixed with chow and administered orally from 1 day before the phosphate-buffered saline or OVA challenge. Lung tissues were harvested after challenge and characterized histologically for infiltrating inflammatory cells, mucus-secreting goblet cells, and collagen deposition. Airway hyperresponsiveness was measured using whole-body plethysmography. Tiplaxtinin treatment significantly decreased levels of PAI-1 activity in bronchoalveolar lavage fluids, which indicates successful blockage of PAI-1 activity in the airways. The number of infiltrated inflammatory cells was reduced by tiplaxtinin treatment in the lungs of the OVA-challenged mice. Furthermore, oral administration of tiplaxtinin significantly attenuated the degree of goblet cell hyperplasia and collagen deposition in the airways of the OVA-challenged mice, and methacholine-induced airway hyperresponsiveness was effectively reduced by tiplaxtinin in these animals. This study supports our previous findings that PAI-1 promotes airway remodeling in a murine model of chronic asthma, and suggests that PAI-1 may be a novel target of treatment of airway remodeling in asthma.

Impact of agr Functionality on the Outcome of Patients with Methicillin-Susceptible Staphylococcus aureus Bacteremia.

Dysfunctional accessory gene regulator (agr) is associated with unfavorable outcomes in invasive methicillin-resistant Staphylococcus aureus infections. However, it is unknown whether this association persists in methicillin-susceptible Staphylococcus aureus bacteremia (MSSA-B). This study evaluated the association between agr dysfunction and mortality in patients with MSSA-B. This retrospective cohort study included MSSA-B patients (≥15 years) enrolled from June 2014 to June 2019 and retrospectively collected their demographic and clinical information. Stored causative strains were measured for agr functionality by δ-hemolysin production assays. Among 244 MSSA-B patients, 91 (37.3%) and 153 (62.7%) had dysfunctional and functional agr MSSA-B, respectively. Ninety-day mortality occurred in 18.7% and 17.6% dysfunctional and functional groups, respectively (P = 0.97). Kaplan-Meier analysis showed that mortality due to dysfunctional agr MSSA-B was not significantly higher (P = 0.82). Age, sites, the severity of infection, and comorbidity adjusted hazard ratio (aHR) of the dysfunctional group for 90-day mortality was 1.303 (95% confidence interval [CI], 0.698 to 2.436, P = 0.41). Mortality due to MSSA-B with sequential organ failure assessment (SOFA) scores of 2 to 5 was significantly higher in the dysfunctional group (P = 0.03), and the dysfunctional agr aHR for 90-day mortality was 3.260 (95% CI, 1.050 to 10.118, P = 0.04). The agr dysfunction of causative organisms can have a significant effect on the outcomes of MSSA-B in patients with moderate severity (SOFA scores 2 to 5). IMPORTANCE Few studies have examined the association between methicillin-susceptible Staphylococcus aureus (MSSA) infection and accessory gene regulator (agr) functionality. We evaluated the association between agr dysfunction and mortality in patients with MSSA bacteremia. Dysfunctional agr is associated with lower survival in MSSA bacteremia patients with moderately severe sequential organ failure assessment (SOFA) scores of 2 to 5. We found that the agr functionality of causative organisms may have an effect on patients' outcomes in MSSA like in methicillin-resistant S. aureus.

Transduced human PEP-1-heat shock protein 27 efficiently protects against brain ischemic insult.

Reactive oxygen species contribute to the development of various human diseases. Ischemia is characterized by both significant oxidative stress and characteristic changes in the antioxidant defense mechanism. Heat shock protein 27 (HSP27) has a potent ability to increase cell survival in response to oxidative stress. In the present study, we have investigated the protective effects of PEP-1-HSP27 against cell death and ischemic insults. When PEP-1-HSP27 fusion protein was added to the culture medium of astrocyte and primary neuronal cells, it rapidly entered the cells and protected them against cell death induced by oxidative stress. Immunohistochemical analysis revealed that, when PEP-1-HSP27 fusion protein was intraperitoneally injected into gerbils, it prevented neuronal cell death in the CA1 region of the hippocampus in response to transient forebrain ischemia. Our results demonstrate that transduced PEP-1-HSP27 protects against cell death in vitro and in vivo, and suggest that transduction of PEP-1-HSP27 fusion protein provides a potential strategy for therapeutic delivery in various human diseases in which reactive oxygen species are implicated, including stroke.

Risk factors for fractures of the proximal humerus: results from the EPIDOS prospective study.

Fracture of the proximal humerus is one of the most frequent fractures attributable to osteoporosis; yet, it has seldom been studied. Two types of factors (related to bone fragility and falls) were evaluated to identify risk factors for proximal humerus fractures as well as to examine possible interactions between them. Subjects were 6901 white women aged > or =75 years and all participated in the EPIDOS study of risk factors for osteoporotic fractures (France, 1992-1998). The baseline examination included measurements of femoral neck bone mineral density (BMD) and calcaneal ultrasound parameters (speed of sound [SOS] and broadband ultrasound attenuation [BUA]), a functional clinical examination, and completing a questionnaire on health status and lifestyle. During a mean of 3.6 (0.8) years of follow-up, 165 women had a humeral fracture. Using multivariate Cox regression models, we identified three predictors related to bone fragility-low BMD (relative risk [RR] = 1.4; 95% CI, 1.1-1.7), low SOS (RR = 1.3; 95% CI, 1.0-1.6), and maternal history of hip fracture (RR = 1.8; 95% CI, 1.0-3.0)-and four fall-related predictors-a previous fall (RR = 3.0; 95% CI, 1.5-6.1), a low level of physical activity (RR = 2.2; 95% CI, 1.1-4.4), impaired balance (RR = 1.8; 95% CI, 1.1-2.9), and pain in lower limb extremity (RR = 1.4; 95% CI, 1.0-2.1). The effect of these fall-related predictors varied according to the BMD level; they were significantly associated with proximal humerus fractures in women with osteoporosis (BMD T score < -2.5) but not in nonosteoporotic women. The incidence of proximal humerus fracture in women with osteoporosis and a low fall risk score (5.1 per 1000 woman-years) was only slightly higher than in nonosteoporotic women (4.6 per 1000 woman-years) and similar to the incidence in women without osteoporosis but a high fall risk score (5.3 per 1000 woman-years). On the other hand, the incidence in women who had both types of risk factors was more than two times higher (12.1 per 1000 woman-years) than in women with only one of the two risk factors. These results suggest that women who have both types of risk factors should receive the highest priority for prevention.

Current results of the surgical management of acoustic neuroma.

A retrospective analysis of 162 consecutive cases in 160 patients who underwent microsurgical resection of vestibular schwannomas between October 1995 and June 2001 was undertaken to compare the results with those of other treatment modalities. Patient hospital records, operative video pictures, neuroimaging studies, audiograms, and follow-up data were reviewed. The mean follow-up period was 24 months.There were 34 small (<1.5 cm), 92 medium (1.5-3 cm), and 36 (>3 cm) large tumors. Six were recurrent tumors. Gross total resection was accomplished in all 34 small tumors and 92 medium tumors but only in 50% of the large tumors. Among the 126 small and medium tumors, the facial nerve was saved anatomically in 124 patients. On long-term follow up, facial function was preserved in 94.4% of all patients. Anatomically, the cochlear nerve was preserved in 55.9% of the small and 20.7% of the medium tumors. Function was preserved (Gardner-Robertson class 1 and 2) in 25% of the small and in 19.4% of the medium tumors. Cerebrospinal leakage was present in 10.5%, meningitis in 9.9%, wound infection in 3.7%, and hematoma or contusion in 2.5%. Only one patient died (mortality rate 0.6%). Our data reflect that surgical removal should be the standard management for acoustic tumors, particularly for large and medium tumors, and can be accomplished with acceptable complication rates.

Glyceollins, a novel class of soy phytoalexins, inhibit angiogenesis by blocking the VEGF and bFGF signaling pathways.

SCOPE: Glyceollins are a novel class of soybean phytoalexins with potential cancer-preventive and antiestrogenic effects. The angiogenic cascade during tumor development consists of the release of angiogenic factors and binding of angiogenic factors to receptors on endothelial cells to activate downstream signaling pathways. However, the potential medicinal value of glyceollins, especially in antiangiogenesis, remains unexplored. METHODS AND RESULTS: Here, we investigated the antiangiogenic activity of glyceollins and their underlying mechanisms. Glyceollins inhibited vascular endothelial growth factor (VEGF) or basic fibroblast growth factor (bFGF) induced in vitro angiogenic activity. Glyceollins inhibited VEGF receptor-2 or FGF receptor-1 activity and their downstream signaling pathways such as extracellular regulated kinase 1/2, c-Jun N-terminal kinase, as well as p38 mitogen-activated protein kinase and focal adhesion kinase induced by VEGF or bFGF. Glyceollins significantly suppressed VEGF receptor-2 kinase activity assayed by the ELISA. Glyceollins significantly attenuated in vivo and ex vivo microvessel development in a dose-dependent manner and tumor growth by suppressing microvessel density in Lewis lung carcinoma (LLC) mouse xenograft. CONCLUSION: Thus, glyceollins, elicited ingredients of soy source, target the signaling pathways mediated by VEGF or bFGF, providing new perspectives into potential therapeutics for preventing and treating hypervascularized diseases including cancer.

Effective biochemical decomposition of chlorinated aromatic hydrocarbons with a biocatalyst immobilized on a natural enzyme support.

The enzymatic decomposition of 4-chlorophenol metabolites using an immobilized biocatalyst was investigated in this study. Catechol 1,2-dioxygenase for ortho ring cleavage obtained via cloning of the corresponding gene cphA-I from Arthrobacter chlorophenolicus A6 was overexpressed and purified. It was found that the cphA-I enzyme could catalyze the degradation of catechol, 4-chlorocatechol, and 3-methylcatechol. The expressed enzyme was immobilized onto a natural enzyme support, fulvic acid-activated montmorillonite. The immobilization yield was as high as 63%, and the immobilized enzyme maintained high substrate utilization activity, with only a 15-24% reduction in the specific activity. Kinetic analysis demonstrated marginal differences in νmax and KM values for the free and immobilized enzymes, indicating that inactivation of the immobilized enzyme was minimal. The immobilized enzyme exhibited notably increased stability against changes in the surrounding environment (temperature, pH, and ionic strength). Our results provide useful information for the effective enzymatic biochemical treatment of hazardous organic compounds.

Synthesis and in vitro antibacterial activity of novel 3-azabicyclo[3.3.0]octanyl oxazolidinones.

We synthesized a series of oxazolidinone-type antibacterials in which morpholine C-ring of linezolid has been modified by substituted 3-azabicyclo[3.3.0]octanyl rings. Acetamide or 1,2,3-triazole heterocycle was used as C-5 side chain of oxazolidinone. The resulting series of compounds was then screened in vitro against panel of susceptible and resistant Gram-positive, Gram-negative bacteria, and Mycobacterium tuberculosis (Mtb). Several analogs in this series exhibited potent in vitro antibacterial activity comparable or superior to linezolid against the tested bacteria. Compounds 10a, 10b, 11a, and 15a displayed highly potent activity against M. tuberculosis. Selected compound 10b showed good human microsomal stability and CYP-profile, and showed low activity against hERG channel.

Pharmacokinetic scaling of SJ-8029, a novel anticancer agent possessing microtubule and topoisomerase inhibiting activities, by species-invariant time methods.

This study examined the pharmacokinetic disposition of SJ-8029, a novel anticancer agent possessing microtubule and topoisomerase inhibiting activities, in mice, rats, rabbits and dogs after i.v. administration. The serum concentration-time curves of SJ-8029 were best described by tri-exponential equations in all these animal species. The mean Cl, V(ss) and t(1/2) were 0.3 l/h, 0.1 l and 63.2 min in mice, 1.5 l/h, 1.6 l and 247.7 min in rats, 13.8 l/h, 39.6 l and 245.9 min in rabbits, and 29.2 l/h, 44.6 l and 117.4 min in dogs, respectively. Based on animal data, the pharmacokinetics of SJ-8029 were predicted in humans using simple allometry and also by several species-invariant time transformations using kallynochron, apolysichron and dienetichron times. The human pharmacokinetic parameters of Cl, V(ss) and t(1/2) predicted by the simple allometry and various species-invariant time methods were 50.4-145.0 l/h, 369.0-579.8 l and 242.0-1448.3 min, respectively. These preliminary parameter values may be useful in designing early pharmacokinetic studies of SJ-8029 in humans.