DataSheet_1_In Vivo Activation and Pro-Fibrotic Function of NF-κB in Fibroblastic Cells During Pulmonary Inflammation and Fibrosis Induced by Carbon Nanotubes.pdf

Exposure to insoluble particles in the lung elicits inflammatory responses that eliminate deposited particulates and repair damaged tissue. Overzealous or prolonged responses lead to chronic conditions, such as fibrosis and malignancy, which are frequently progressive and refractory to drug therapy leading to high rates of disability and mortality. The molecular events underlying the progression of lung inflammation to chronic pathology, in particular, the conversion to fibrosis, remain poorly understood. Fibrogenic multi-walled carbon nanotubes (MWCNTs) have been shown to stimulate prominent acute inflammation that evolves into chronic lesions characterized by chronic inflammation, interstitial fibrosis, and granulomas in mouse lungs. In this communication, we examined the in vivo activation of nuclear factor-κB (NF-κB) signaling in fibroblastic cells during the inflammatory and fibrotic progression induced by MWCNTs. Wild-type C57BL/6J male mice were exposed to two fibrogenic MWCNTs (Mitsui XNRI MWNT-7 and long MWCNTs) by pharyngeal aspiration. Both MWCNTs strongly stimulated the nuclear translocation of NF-κB p65 in lung fibroblasts and myofibroblasts during the acute and chronic responses. Phosphorylated NF-κB p65 at serine 276, a marker of NF-κB activation, was markedly induced by MWCNTs in the nucleus of fibroblastic cells. Moreover, two NF-κB-regulated genes encoding pro-fibrotic mediators, tissue inhibitor of metalloproteinase 1 (TIMP1), and osteopontin (OPN), respectively, were significantly induced in lung fibroblasts and myofibroblasts. These results demonstrate that NF-κB is activated to mediate transactivation of pro-fibrotic genes in fibroblastic cells during pulmonary acute and chronic responses to CNTs, providing a mechanistic framework for analyzing gene regulation in pulmonary fibrotic progression through NF-κB signaling.</p

TIMP1 promotes multi-walled carbon nanotube-induced lung fibrosis by stimulating fibroblast activation and proliferation

<p>Pulmonary exposure to multi-walled carbon nanotubes (MWCNTs) may cause fibrosing lesions in animal lungs, raising health concerns about such exposure in humans. The mechanisms underlying fibrosis development remain unclear, but they are believed to involve the dysfunction of fibroblasts and myofibroblasts. Using a mouse model of MWCNT exposure, we found that the tissue inhibitor of metalloproteinase 1 (Timp1) gene was rapidly and highly induced in the lungs by MWCNTs in a time- and dose-dependent manner. Concomitantly, a pronounced elevation of secreted TIMP1 was observed in the bronchoalveolar lavage (BAL) fluid and serum. Knockout (KO) of Timp1 in mice caused a significant reduction in fibrotic focus formation, collagen fiber deposition, recruitment of fibroblasts and differentiation of fibroblasts into myofibroblasts in the lungs, indicating that TIMP1 plays a critical role in the pulmonary fibrotic response to MWCNTs. At the molecular level, MWCNT exposure significantly increased the expression of the cell proliferation markers Ki-67 and PCNA and a panel of cell cycle-controlling genes in the lungs in a TIMP1-dependent manner. MWCNT-stimulated cell proliferation was most prominent in fibroblasts but not myofibroblasts. Furthermore, MWCNTs elicited a significant induction of CD63 and integrin β1 in lung fibroblasts, leading to the formation of a TIMP1/CD63/integrin β1 complex on the surface of fibroblasts <i>in vivo</i> and <i>in vitro</i>, which triggered the phosphorylation and activation of Erk1/2. Our study uncovers a new pathway through which induced TIMP1 critically modulates the pulmonary fibrotic response to MWCNTs by promoting fibroblast activation and proliferation via the TIMP1/CD63/integrin β1 axis and ERK signaling.</p

Suppression of basal and carbon nanotube-induced oxidative stress, inflammation and fibrosis in mouse lungs by Nrf2

<p>The lungs are susceptible to oxidative damage by inhaled pathogenic agents, including multi-walled carbon nanotubes (MWCNT). The nuclear factor erythroid 2-related factor 2 (Nrf2) has been implicated in regulating the body’s defense against oxidative stress. Here, we analyzed the function of Nrf2 in the lungs. Under a basal condition, Nrf2 knockout (KO) mice showed apparent pulmonary infiltration of granulocytes, macrophages and B and T lymphocytes, and elevated deposition of collagen fibers. Exposure to MWCNT (XNRI MWNT-7, Mitsui, Tokyo, Japan) by pharyngeal aspiration elicited rapid inflammatory and fibrotic responses in a dose (0, 5, 20 and 40 μg) and time (1, 3, 7 and 14 d)-dependent manner. The responses reached peak levels on day 7 post-exposure to 40 μg MWCNT, evidenced by massive inflammatory infiltration and formation of inflammatory and fibrotic foci, which were more evident in Nrf2 KO than wild-type (WT) lungs. At the molecular level, Nrf2 protein was detected at a low level under a basal condition, and was dramatically increased by MWCNT in WT, but not Nrf2 KO, lungs. Activation of Nrf2 was inversely correlated with induced expression of fibrosis marker genes and profibrotic cytokines. Furthermore, the levels of ROS and oxidative stress were remarkably higher in Nrf2 KO than WT lungs under a physiological condition, and were dramatically increased by MWCNT, with the increase significantly more striking in KO lungs. The findings reveal that Nrf2 plays an important role in suppressing the basal and MWCNT-induced oxidant production, inflammation and fibrosis in the lungs, thereby protecting against MWCNT lung toxicity.</p

Synthesis, Structures, and Norbornene Polymerization Behavior of Imidazo[1,5‑<i>a</i>]pyridine-sulfonate-Ligated Palladacycles

Two imidazo­[1,5-a]­pyridine-sulfonate proligands, L1 and L2, were synthesized in five-step reactions. Treatment of the proligands with palladacycles {[Pd­(OAc)­(8-Me-quin-H)]2, [Pd­(dmba)­(μ-Cl)]2, and [Pd­(o-acetanilido)­(μ-trifluoroacetato)]2} yielded the desired five-membered C­(sp3),N-chelated (Pd1, Pd2), C­(sp2),N-chelated (Pd3, Pd4), and six-membered C­(sp2),O-chelated (Pd5, Pd6) palladacycles, respectively. All these complexes were fully characterized by 1H and 13C NMR, IR, high-resolution mass spectrometry, and elemental analysis. The molecular structures of complexes Pd1, Pd2, Pd4, and Pd5 were determined by single-crystal X-ray diffraction analysis. In the presence of MAO or Et2AlCl, Pd1–Pd6 exhibited activities toward the addition polymerization of norbornene which decreased in the order Pd6 > Pd5 > Pd4 > Pd2 > Pd3 > Pd1. The Pd1–Pd6/MAO catalytic system showed high thermal stability and reached the highest activity at 100 °C (6.0 × 107 g of PNB (mol of Pd)−1 h–1 with 99.9% conversion). In the presence of Et2AlCl with low loading (100 equiv), Pd5 and Pd6 exhibited high activities (up to 2.9 × 107 g of PNB (mol of Pd)−1 h–1 with 96.5% conversion). It was demonstrated that the structures of palladacycles and the substituents on the ligands significantly affected the activities of these complexes

Real Time Evaluation of Composition and Structure of Concanavalin A Adsorbed on a Polystyrene Surface

In situ qualitative and quantitative evaluations of adsorbed submonolayers and multilayers of the protein Concanavalin A (ConA) on a polystyrene surface were carried out by attenuated total reflection infrared spectroscopy. The influence of pH and adsorption time on the composition and structure of the adsorbed protein layers was investigated by comparison of the experimental spectra with simulated spectra of hypothetical multilayer systems with the assumed composition, thickness, and structure. This methodology allows the differentiation of observed spectral changes that result from pure optical effects, associated with the passing of an incident beam through the multilayer system, from the chemical and structural changes caused by physicochemical interactions of proteins with polymer surfaces. This represents significant progress since small variations in the band positions or intensities of amide I and amide II infrared absorbance bands have an important interpretation consequence. The applied methodology significantly reduces the misinterpretation of recorded spectra of protein layers and is rewarded by a deep insight of the structure and composition of the samples. The composition, structure, and kinetics of the adsorption of ConA and hydration level of the adsorbed layers were evaluated in detail. Competitive adsorption of bovine serum albumin on pre-adsorbed ConA layers also was investigated to characterize the ConA surface distribution. Parallel studies using X-ray photoelectron spectroscopy support the conclusions drawn from infrared spectroscopic investigations on ConA molecular distributions at the polymer surface. Two-step models that describe ConA submonolayer formation at pH 4.8 and multilayer formation at pH 7.8 are proposed

Fig 4 -

The calibration curve indicates good consistency between training set(A) and validation set(B).</p

Fig 3 -

The ROC analyses for the predictive model in training set(A) and validation set(B).</p

Nomogram for predicting drug shock-outs.

IntroductionReasons for drug shortages are multi-factorial, and patients are greatly injured. So we needed to reduce the frequency and risk of drug shortages in hospitals. At present, the risk of drug shortages in medical institutions rarely used prediction models. To this end, we attempted to proactively predict the risk of drug shortages in hospital drug procurement to make further decisions or implement interventions.ObjectivesThe aim of this study is to establish a nomogram to show the risk of drug shortages.MethodsWe collated data obtained using the centralized procurement platform of Hebei Province and defined independent and dependent variables to be included in the model. The data were divided into a training set and a validation set according to 7:3. Univariate and multivariate logistic regression were used to determine independent risk factors, and discrimination (using the receiver operating characteristic curve), calibration (Hosmer-Lemeshow test), and decision curve analysis were validated.ResultsAs a result, volume-based procurement, therapeutic class, dosage form, distribution firm, take orders, order date, and unit price were regarded as independent risk factors for drug shortages. In the training (AUC = 0.707) and validation (AUC = 0.688) sets, the nomogram exhibited a sufficient level of discrimination.ConclusionsThe model can predict the risk of drug shortages in the hospital drug purchase process. The application of this model will help optimize the management of drug shortages in hospitals.</div

Univariate and multivariate analysis of drug shortages in the training set.

Univariate and multivariate analysis of drug shortages in the training set.</p