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Urinary stone is conceptualized as a chronic metabolic disorder punctuated by symptomatic stone events. It has been shown that the occurrence of calcium oxalate monohydrate (COM) during stone formation is regulated by crystal growth modifiers. Although crystallization inhibitors have been recognized as a therapeutic modality for decades, limited progress has been made in the discovery of effective modifiers to intervene with stone disease. In this study, we have used metabolomics technologies, a powerful approach to identify biomarkers by screening the urine components of the dynamic progression in a bladder stone model. By in-depth mining and analysis of metabolomics data, we have screened five differential metabolites. Through density functional theory studies and bulk crystallization, we found that three of them (salicyluric, gentisic acid and succinate) could effectively inhibit nucleation in vitro. We thereby assessed the impact of the inhibitors with an EG-induced rat model for kidney stones. Notably, succinate, a key player in the tricarboxylic acid cycle, could decrease kidney calcium deposition and injury in the model. Transcriptomic analysis further showed that the protective effect of succinate was mainly through anti-inflammation, inhibition of cell adhesion and osteogenic differentiation. These findings indicated that succinate may provide a new therapeutic option for urinary stones.

An interpatient classification method using the second lead for the Association for the Advancement of Medical Instrumentation (AAMI) standard is introduced. This study includes three parts. In the first part, the fuzzy matching algorithm is used to locate the key waveform points of Electrocardiogram (ECG) data. In the second part, the feature engineering algorithm is used to filter the extracted data sets. In the third part, the random forest model is carried out to realize the five classifications of heart disease by the selected features. The final precision, recall, and F1-score are 91%, 89%, and 90%, respectively.

This study proposes a personal healthcare system to monitor, identify ectopic heartbeats, and provide services to patients with heart diseases. The proposed system includes three subsystems: (1) an electrocardiography (ECG) signal acquisition device to acquire ECG signals accurately; (2) an Android-based smartphone to display the actual ECG waveform and upload the signal to the Web platform for analysis of cardiac disorder events; and (3) a Web platform for feature selection and arrhythmia classification with the support vector machine algorithm. The classification algorithm can identify three types of heartbeats, namely, normal, atrial premature beat, and premature ventricular contraction. The average sensitivity, specificity, and accuracy of the classification algorithm over the training set are 98.5%, 96.7%, and 97.7%, respectively.

Power efficiency and stability are critical for wearable ECG application. The paper presents a Pulse-Width-Modulation (PWM) voltage type DC-DC converter, which has a small area of 0.32 mm2 on active chip, output voltage range of 0.8V to 3.5V, output power up to 7.5 mW, and peak efficiency of 95.8%. The simulation results show that the output voltage of the system is 1.8V, the output current is 4mA, the ripple voltage is 13mV and the efficiency is 86.1%, which meets the requirements of a presented system. The circuit will be fabricated by the SMIC in the near future with $0.18\ \mu\mathrm{m}$ Bipolar-CMOS-DMOS (BCD) process.

Lignin only accounts for about 6% of total mass in tobacco stem, but it influences the harmful substances in the side stream smoke of cigarette in a significant way. Traditional researches focus only on the determination of lignin content. In the present work, we investigate four typical imidazolium-based ionic liquids for efficient extraction of lignin under mild conditions and 1-ethyl-3-methylimidazolium diethylphosphate ([Emim][DEP]) shows the best results. The pretreatment of stem using water at 80 °C for 30 min can not only remove most of the sugars but also loose the microfibers. The extractive rate of lignin reaches 85.38% at 150 °C for 4 h and the purity of lignin is 90.21%.

In the present work, we reported a novel route for the conversion of lignocellulosic biomass (sawdust) to a high-value chemical of benzoic acid under atmospheric pressure. The transformation involved the catalytic pyrolysis of sawdust into aromatics, the decomposition of heavier alkylaromatics to toluene, and the liquid-phase oxidation of toluene-rich aromatics to benzoic acid. The production of the desired benzoic acid from the sawdust-derived aromatics, with the benzoic acid selectivity of 85.1 C-mol% and nearly complete conversion of toluene, was achieved using the MnO2/NHPI catalyst at 100 °C for 5 h. The influence of adding methanol on the catalytic conversion of sawdust to the core intermediate of toluene was also investigated in detail.

The directional production of benzene is achieved by the current-enhanced catalytic conversion of lignin. The synergistic effect between catalyst and current promotes the depolymerization of lignin and the selective recombinant of the functional groups in the aromatic monomers. A high benzene yield of 175 gbenzene/kglignin was obtained with an excellent selectivity of 92.9 C-mol%. The process potentially provides a promising route for the production of basic petrochemical materials or high value-added chemicals using renewable biomass.

The study proposed a switched-capacitor delta-sigma modulator with single-loop and third-order structures for wearable electrocardiogram acquisition device for low-power and high-precision applications. Several techniques, including chain of integrators with weighted feedforward structure, gain-enhanced current-mirror push-pull operational transconductance amplifier, and dynamic comparator with a set-reset latch are adopted to improve the signal-to-noise ratio (SNR) and reduce the power consumption. Simulation results shows that the SNR and the corresponding effective number of bits are 104.2 dB and 17.02 bits, respectively, which fulfill the requirements of an presented system. The circuit will be fabricated by the SMIC using a 0.18 μm general purpose process in the near future.

The AlCl3/Ti co-doped 4MgH2-Li3AlH6 has been successfully synthesized using solid ball-milled method. The effect of AlCl3/Ti and different preparation conditions on reversible hydrogen storage of 4MgH2-Li3AlH6 were investigated. It was found that the formed Al3Ti and Al can improve the de/rehydriding performance. The onset temperature of the system dehydrogenation decreased to 58 °C, and it can release hydrogen of 7.2 wt.% at 400 °C. The hydriding of the system was found that the adsorption rate greatly increased from 0.02 to 0.35 wt.% min-1. The activation energy (Ea) of MgH2 dehydrogenation decreased from 147 to 113.7 kJ mol-1 by adding additive AlCl3 and Ti. The AlCl3/Ti improved the thermodynamic and kinetic performance of the 4MgH2-Li3AlH6 composite, which was attributed to the high catalytic activity of Al3Ti and Al.

Acute respiratory distress syndrome is an often fatal disease that develops after acute lung injury and trauma. How released tissue damage signals, or alarmins, orchestrate early inflammatory events is poorly understood. Herein we reveal that IL-33, an alarmin sequestered in the lung epithelium, is required to limit inflammation after injury due to an unappreciated capacity to mediate Foxp3(+) Treg control of local cytokines and myeloid populations. Specifically, Il33(–/–) mice are more susceptible to lung damage–associated morbidity and mortality that is typified by augmented levels of the proinflammatory cytokines and Ly6C(hi) monocytes in the bronchoalveolar lavage fluid. Local delivery of IL-33 at the time of injury is protective but requires the presence of Treg cells. IL-33 stimulates both mouse and human Tregs to secrete IL-13. Using Foxp3(Cre) × Il4/Il13(fl/fl) mice, we show that Treg expression of IL-13 is required to prevent mortality after acute lung injury by controlling local levels of G-CSF, IL-6, and MCP-1 and inhibiting accumulation of Ly6C(hi) monocytes. Our study identifies a regulatory mechanism involving IL-33 and Treg secretion of IL-13 in response to tissue damage that is instrumental in limiting local inflammatory responses and may shape the myeloid compartment after lung injury.

Objective: To analysis of risk factors for length of stay in intensive care unit and survival outcome from premature infants with premature rupture of membranes (PROM). Methods: A retrospective study including 189 singleton pregnant women with PPROM at 28– 34 gestational weeks was conducted. Gestational age at delivery, route of delivery, oligohydramnios, maternal age and maternal complications including gestational diabetes, preeclampsia and chorioamnionitis, newborn birth weight, gender, premature retinopathy (ROP), necrotizing enterocolitis (NEC), sepsis, fetal growth retardation (FGR), intracranial hemorrhage (ICH), bronchopulmonary dysplasia (BPD), respiratory distress syndrome (RDS), primary pulmonary hypertension (PPH), congenital cardiac disease (CCD), patent ductus arteriosus (PDA) and use of betamethasone were used to predict neonatal outcomes in terms of length of stay in the ICU and survival. Results: The preeclampsia, low birth weight, ROP, CCD, BPD, and PDA were significant confounders for length of stay in the ICU. Among them, low birth weight was the most powerful confounder for prolongation of the ICU stay (P

Motion artifact removal is one of the most important issues in portable ECG (electrocardiogram) monitoring. Motion artifacts are usually accompanied by significant baseline drift. Adaptive filtering has been used to filter motion artifacts and baseline drift. The 3-axis acceleration signal is selected as the noise reference signal. Results show that motion artifacts and baseline drift have been removed in filtered ECG, and the QRS complex of the filtered ECG has clearly appeared.

Although extracorporeal membrane oxygenation (ECMO) has been used frequently as a bridge to primary lung transplantation, active centers are conservative with this approach in patients requiring redo lung transplantation. We report the use of extracorporeal carbon dioxide removal, using the Hemolung respiratory assist system, as a prolonged bridge to lung transplantation, and the first use of the Hemolung as a bridge to redo lung transplantation. Hemolung support improved the patient's clinical status and allowed redo lung transplantation.

IL-33, a relatively new member of the IL-1 cytokine family, plays a crucial role in allergic inflammation and acute lung injury. Long form ST2 (ST2L), the receptor for IL-33, is expressed on immune effector cells and lung epithelia and plays a critical role in triggering inflammation. We have previously shown that ST2L stability is regulated by the ubiquitin-proteasome system; however, its upstream internalization has not been studied. In this study, we demonstrate that glycogen synthase kinase 3β (GSK3β) regulates ST2L internalization and IL-33 signaling. IL-33 treatment induced ST2L internalization, and an effect was attenuated by inhibition or downregulation of GSK3β. GSK3β was found to interact with ST2L on serine residue 446 in response to IL-33 treatment. GSK3β binding site mutant (ST2LS446A) and phosphorylation site mutant (ST2LS442A) are resistant to IL-33–induced ST2L internalization. We also found that IL-33 activated focal adhesion kinase (FAK). Inhibition of FAK impaired IL-33–induced GSK3β activation and ST2L internalization. Furthermore, inhibition of ST2L internalization enhanced IL-33–induced cytokine release in lung epithelial cells. These results suggest that modulation of the ST2L internalization by FAK/GSK3β might serve as a unique strategy to lessen pulmonary inflammation.

A three-lead electrocardiogram acquisition system is constructed with an analog front-end circuit (ADS1293), a microprocessor (MSP430X), and a personal phone or computer. The acquired three-lead (namely, Lead I, Lead II, and Lead V1) ECG data can not only be displayed on a personal computer by a MATLAB simulator but can also be stored in the cloud.

Systemic sclerosis (SSc), or scleroderma, is a heterogeneous and complex autoimmune disease characterized by varying degrees of skin and organ fibrosis and obliterative vasculopathy. The disease results in significant morbidity and mortality, and to date, available treatments are limited. Lung involvement is the leading cause of death of patients with SSc. Over the past year, significant advances have been made in our understanding of SSc-associated lung disease, and this review attempts to encapsulate these most recent findings and place them in context.We divide our discussion of the most recent literature into the following: first, clinical aspects of SSc lung management, including classification, imaging, biomarkers, and treatment; second, promising new animal models that may improve our ability to accurately study this disease; and third, studies that advance or change our understanding of SSc lung disease pathogenesis, thereby raising the potential for new targets for therapeutic intervention.Recent advances have resulted in a better understanding of SSc-associated lung disease, the development of new in-vivo models for exploring disease pathogenesis, and the identification of potential novel targets for the development of therapies.

This paper presents a bandgap reference and an output-capacitorless LDO regulator with adaptive power transistors. The bandgap reference consists of a current reference circuit, a bipolar transistor and proportional-to-absolute-temperature (PTAT) voltage generators. The proposed LDO improves load transient and light load efficiency by permitting the regulator to transform itself between 2-stage and 3-stage topologies, depending on the load current condition. Cadence simulation with SMIC 0.18 μm process shows that the bandgap reference generates a reference voltage 569 mV and the quiescent current is only 0.23 μA, the proposed LDO generates an output voltage 1 V, the quiescent current is 0.88 μA (including bandgap reference) at no-load condition, the undershoot /overshoot voltage is 187 mV/152 mV and the settling time is 5 μs as load current suddenly changes from 0 to 100 mA, or vice versa.

This paper presents a capacitor-less CMOS low dropout regulator (LDO) with a push-pull class AB amplifier, and a fast transient controller to achieve a better transient response. The undershoot/overshoot voltage and the settling time are effectively reduced. Through the theoretical analysis of the circuit, cadence simulation with SMIC 0.18μm process and under the condition of the input voltage range 1.4~4 V shows the output voltage is 1.2 V, with the fast controller the total quiescent current is 8.2 μA, the undershoot /overshoot voltage is 97 mV/47 mV and the settling time is 0.3 μs as load current suddenly changes from 1 to 100 mA, or vice versa. Compared with this paper without fast transient controller, the undershoot voltage, the overshoot voltage and the settling time are enhanced by 30%, 64% and 80%, respectively.

Conversion of biomass to chemicals or fuels under mild condition is still a challenge. As a platform molecule for chemicals and fuels, levulinic acid (LA) has been prepared by liquefaction of biomass at high pressure. In order to carry out the conversion from wheat straw to LA at atmosphere pressure, continuous extraction of the reactive system by an organic solvent with a higher density than that of water was utilized for degradation of pretreated biomass. Yields of LA were measured by means of gas chromatography-mass spectrometry and nuclear magnetic resonance. The results revealed that a maximum yield of 30.66% of LA can be obtained from wheat straw. In addition, the effects of biomass pretreated conditions on the LA conversion have been studied. The study provides a new route to convert biomass to valuable chemicals at atmosphere pressure.

This paper describes in detail an electrocardiogram (ECG) acquisition system comprised of ADS1291, MSP430 and CC2541. ADS1291 amplifies weak analog ECG signal and changes it into digital signals, and then sends it to back-end system and displays it. Patient can use it to acquire ECG signals effectively and simply.

Although advances in surgical technique, immunosuppressive regimens, and medical management have led to improved survival and quality of life after solid organ transplantation, infection continues to represent a major cause of morbidity and mortality in transplant recipients. Immunosuppressive therapy after transplantation compromises cell-mediated immunity in particular, leaving the patient at risk for opportunistic as well as routine community-acquired infections. Mycobacterial infection is a rare but important complication of solid organ transplantation, presenting significant risk to the patient and challenges in terms of treatment. The available literature consists predominantly of case reports and institutional experiences. This article examines both Mycobacterium tuberculosis and nontuberculous mycobacterial infection in the transplant setting.

Due to the difficulties in diagnosing arrhythmia and patients' enthusiasm to attach greater attention to their health, traditional electrocardiogram (ECG) monitoring is facing many challenges. In addition, medical resource distribution problems are particularly obvious. Therefore, the ECG network system for medical resource comprehensive application is proposed in this paper. It is conducive to increasing the cure rate significantly and reducing imbalance of distribution in medical resources. The network system can also effectively improve medical services. The initial implementation proves that ECG network system proposed in this paper is feasible and reasonable.

The effect of sodium chloride on activity and stability of polyphenol oxidase (PPO, EC1.14.18.1) from Fuji apple was investigated. Partially purified PPO was activated with 0.17–3.4 M NaCl. Purified PPO stored with NaCl was stable for 2 months at 4C. PPO in the presence of NaCl (0–5.1 M) was scanned by Differential UV from 240 to 300 nm and spectral analysis showed that there were conformational changes as a function of NaCl concentration.

Acetaminophen toxicity is the most common cause of acute liver failure in the United States and Europe. Although much is known about the metabolism of acetaminophen, many questions remain regarding the pathogenesis of liver injury. In this study, we examined the role of lipopolysaccharide-binding protein (LBP), a protein important in mediating cellular response to lipopolysaccharides, by using LBP wild-type and knockout (KO) mice. We found that LBP KO mice were protected from acetaminophen-induced hepatotoxicity. At 350 mg/kg of acetaminophen, LBP KO mice had significantly less liver injury and necrosis than wild-type mice. Repletion studies in LBP KO mice using an LBP-adenoviral construct resulted in significantly more hepatic injury and necrosis after acetaminophen exposure compared with mice receiving the control adenoviral construct. In conclusion, LBP KO mice are protected from toxicity with a decrease in hepatic necrosis following acetaminophen challenge. This suggests a novel role for LBP in modulating acetaminophen-induced liver injury. Supplementary material for this article can be found on the HEPATOLOGY website (http://interscience.wiley.com/jpages/O270-9139/suppmat/index.html).

The resection of primary and secondary liver tumors has become accepted as the only curative therapy that can be offered to patients with these cancers. Technical advances made over the last two decades have improved the ability of the surgeon to perform these procedures with decreased morbidity. This article reviews hepatic anatomy, the preoperative evaluation of patients and various technical aspects involved in liver resections. The latter includes the role of intraoperative ultrasound and techniques of vascular occlusion and hepatic parenchymal dissection.

Septic complications and the emergence of drug-resistant microbes represent serious risks to patients. Recently, naturally occurring peptides have been discovered that possess potent and broad-spectrum antimicrobial activity. Protegrin-1 is particularly attractive for clinical use in human wounds because, unlike defensins, protegrin-1 retains broad antimicrobial and antifungal activity at physiologic salt concentration and in the presence of serum. The objective of this study was to examine the efficacy of protegrin-1 in killing multiple drug-resistant microbes isolated from human burn patients.For thein vitroexperiment, bilayer radial diffusion was performed comparing standard antibiotics with protegrin-1 on multiple-drug-resistant microbial organisms isolated from infected burn wounds. In vivo, rats received a 20% total body surface area partial-thickness burn by immersion in 60 degrees C water for 20 secs followed by wound seeding with 106 colony forming units of Silvadene-resistant Pseudomonas aeruginosa.University of Michigan research laboratory.Adult, male Sprague-Dawley rats.Rats were randomized into three groups: those receiving synthetic protegrin-1, acetic acid (carrier), or gentamicin (positive control). Protegrin-1 was administered by topical application or intradermal injection. Wound tissues were harvested aseptically at different time points for quantitative bacterial counts.In vivo and in vitro experiments revealed rapid and significant decreases in bacterial counts for protegrin-1-treated groups compared with controls.This study shows that protegrin-1 potentially may be used as an alternative or adjunct therapy to standard agents used to treat wound infections.

Recently, developmental neurobiologists have identified multipotential stem cells in the mature nervous systems of vertebrates. The identification of these progenitor cells, which perdure into adulthood, may have important implications for how we think about brain tumor pathology, as we must now consider the possibility that some tumors may arise from these multipotential precursor cells. Currently, neuropathological classification systems are based primarily upon morphological criteria. While these classification systems provide important information regarding prognosis, they have thus far provided little insight into how, from where, and why any given tumor arises. Even prior to having the knowledge that stem cells are present in the adult central nervous system (CNS), several investigators began to explore the idea that antigenic markers, which have proven invaluable to our understanding of what constitutes the normal repertoire of behaviors and lineages of CNS cells, may provide insights into the basic biology of brain tumors. In the following exegesis, we first highlight two papers that review some of the exciting recent advances in developmental neurobiology and point out what implications this may have on our understanding of brain malignancies. We then present several studies in which the investigators sought to test the hypothesis that the antigenic markers that have proved so useful for the developmental neurobiologist may also prove useful in trying to understand the similarities and differences in various categories of brain tumors. While none of these studies demonstrate definitively that brain tumors can or do arise from the aberrant behavior of CNS stem cells, they do provide enough circumstantial evidence that this may be the case to warrant further serious investigation of this hypothesis.

Polar residues within the transmembrane domains (TMs) of G protein-coupled receptors have been implicated to be important determinants of receptor function. We have identified mutations at two polar sites in the TM regions of the rat parathyroid hormone (PTH)/PTH-related peptide receptor, Arg-233 in TM 2 and Gln-451 in TM 7, that caused 17-200-fold reductions in the binding affinity of the agonist peptide PTH-(1-34) without affecting the binding affinity of the antagonist/partial agonist PTH-(3-34). When mutations at the TM 2 and TM 7 sites were combined, binding affinity for PTH-(1-34) was restored to nearly that of the wild type receptor. The double mutant receptors, however, were completely defective in signaling cAMP or inositol phosphate production in response to PTH-(1-34) agonist ligand. The results demonstrate that Arg-233 and Gln-451 have important roles in determining agonist binding affinity and transmembrane signaling. Furthermore, the finding that residues in TM 2 and TM 7 are functionally linked suggests that the TM domain topology of the PTH/PTH-related peptide receptor may resemble that of receptors in the rhodopsin/beta-adrenergic receptor family, for which structural and mutagenesis data suggest interactions between TMs 2 and 7.

Non-lymphoid tissue-resident CD4+ Foxp3+ regulatory T (Treg) cells with the capacity to modulate non-immunological processes including organismal metabolism and tissue repair have been recently described. Notably, a large fraction of non-lymphoid tissue-resident Treg cells express ST2, the receptor for the tissue-derived cytokine and alarmin, IL-33. However, the relationship between IL-33 and ST2+ Treg cells in quiescent and pathogenic states is only starting to be understood. Using FACS and Foxp3 reporter mice, we demonstrate that ST2+ Treg cells from naïve animals are phenotypically and functionally unique relative to ST2− Treg isolated from the same location. Yet, our studies establish that peripheral and lymphoid tissue ST2+ Treg cells are highly comparable. Interestingly, ST2+ Treg exhibit a high level of aerobic glycolysis, which supports their augmented potential for cytokine secretion, especially that of IL-5, IL-10, and IL-13. IL-5 and IL-13 secretion in response to IL-33 is an innate function of ST2+ Treg cells as it does not require TCR-stimulation. Conversely, ST2+ Treg release of other cytokines, including IL-10, is antigen-restricted and amplified by IL-33. Among Treg subsets, ST2+ Treg cells are the exclusive source of Type 2 cytokines, especially IL-13, which we reveal supports Treg reparative responses. Using mice deficient in IL-33 or Treg cells, we establish that Treg cells and IL-33 are both required to protect mice from lethal lung injury. In line with these data, delivery of IL-13, but not IL-33, at the time of lung injury rescues Treg-depleted mice. This study establishes that ST2+ Treg cells possess an innate capacity for cytokine production that is crucial to tissue repair and inflammation control.

Background: We previously demonstrated an essential role for lipopolysaccharide binding protein (LBP) in the pulmonary immune response to Gram-negative bacterial infection. LBP knockout mice had significantly higher mortality, greater rates of bacteremia, and higher counts of viable bacteria in their lungs at killing than did wild-type controls. We postulate that systemic LBP gene therapy will reconstitute a protective innate immune response in LBP knockout mice and that overexpression of LBP in wild-type mice may offer a survival advantage. Methods: Twelve-to 16-week-old female C57BL/6 wild-type mice and age-matched LBP knockout mice were given 5 × 109 plaque-forming units of recombinant adenovirus containing either the gene for LBP or the irrelevant control protein β-galactosidase by tail vein injection; 72 hours later; each mouse was administered 1 × 103 colony-forming units of Klebsiella pneumoniae by intratracheal injection. Results: Administration of LBP by systemic gene therapy to LBP knockout mice improved survival from Klebsiella pneumonia to a level equivalent to or better than that in wild-type mice exposed to the same dose of bacteria (36% versus 25%). Wild-type mice given the LBP gene therapy demonstrated increased 7-day survival from Klebsiella pneumonia when compared with controls treated with β-galactosidase (68% versus 30%; p = 0.03). Conclusion: Systemic gene therapy with intravenous adenoviral vector transfer of LBP significantly improves survival in LBP knockout mice. Overexpression of LBP in wild-type mice improves survival from Klebsiella pneumonia. Raising levels of LBP in the setting of Gram-negative pneumonia may be of therapeutic benefit.

As semiconductor manufacturing moves to the 90nm node and below, shrinking feature sizes and increasing IC complexity have combined to significantly stretch out the time needed to optimize and qualify process anchored OPC models and recipes. Process distortion and non-linearity become non-trivial issues and conspire to reduce the quality of the resulting corrections. Additionally, optimizing the OPC model and recipe on a limited set of test chip designs may not provide sufficient coverage across the range of designs to be produced in the process. Finally, the increased complexity of the transformation of the target pattern into a corrected mask pattern also increases the probability of system lithography errors. Fatal errors (pinch or bridge) or poor CD distribution may still occur. As a result, more than one reticle tape-out cycle is non uncommon to prove models and recipes that approach the center of process for a range of designs. In this paper, we describe a full-chip simulation based verification flow using a commercialized product that serves both OPC model and recipe development as well as post OPC verification after production release of the OPC.

Background An altered Kupffer cell (KC) response is thought to be responsible for the characteristic phenotype observed after biliary obstruction: a phenotype marked by a defect in the hepatic reticuloendothelial system and a hypersensitivity to endotoxin. Few studies, however, have directly examined KC function. We have sought to define the specific alterations in function and phenotype that occur in the KC after biliary obstruction. Methods KCs were isolated from female C57BL/6 mice 4 days after a sham or common bile duct ligation (CBDL) operation. Phagocytosis, oxidative burst potential, and intracellular bacterial killing were measured as markers of reticuloendothelial system function. The KC response to endotoxin was assessed by measuring tumor necrosis factor alpha and interleukin 6 levels in the media after stimulation with lipopolysaccharide (LPS) or with LPS plus LPS-binding protein (LBP). Results CBDL KCs demonstrated a significant increase in phagocytic ability and significantly decreased baseline oxidative stress, compared with Shams. The oxidative burst potential, however, was equivalent or higher for CBDL KCs. CBDL KCs also demonstrated increased numbers of viable intracellular bacteria after infection; however, it is unclear if this finding represents impaired intracellular bacterial killing or increased phagocytosis of bacteria. With respect to the KC response to endotoxin, CBDL KCs were found to be less sensitive to the stimulatory effects of LPS alone but were exquisitely sensitive to the effects of LBP. LBP levels were found to be significantly elevated in CBDL animals, and CBDL KCs demonstrated a dose-dependent, exaggerated tumor necrosis factor alpha and interleukin 6 response to LPS administered with LBP. Conclusions KC function is clearly altered after biliary obstruction. Phagocytic ability is actually increased, although the ability of CBDL KCs to kill bacteria within the phagosome remains ill defined. CBDL KCs are exquisitely sensitive to the effects of LBP, and LBP levels are elevated after biliary obstruction. LBP may be responsible for the increased proinflammatory response observed after endotoxin challenge in animals with biliary obstruction.

Lipopolysaccharide (LPS)-binding protein (LBP) greatly facilitates LPS activation of monocytic cells through the CD14 receptor, triggering activation of innate immune responses. An acute phase protein, LBP is produced predominantly by the liver; however, we and others have shown that LBP is produced extrahepatically in multiple locations, including the lung. The importance of LBP in the lung has remained unclear. LBP may make the host more acutely sensitive to LPS and development of septic complications; alternatively, it may be protective, aiding in detection, opsonization, and killing of bacteria. Our objective was to determine the role LBP plays in local pulmonary immune defenses to bacterial challenge. LBP knockout mice and age-matched C57BL/6 wild-type controls were challenged with direct intratracheal inoculation of Klebsiella pneumoniae. We observed a significant increase in mortality, earlier onset of bacteremia, and greater pulmonary bacterial loads in LBP knockout mice compared with controls. Total lung myeloperoxidase (MPO) activity, neutrophil recruitment to the alveolar space, and levels of KC--a chemokine involved in neutrophil recruitment--in bronchoalveolar lavage (BAL) fluid and lung homogenates were found to be significantly diminished in knockout mice compared with controls. Together, our findings suggest that LBP is essential in local pulmonary innate immune responses against bacteria.

Upregulation of CD14 in Kupffer cells has been implicated in the pathogenesis of several forms of liver injury, including alcoholic liver disease. However, it remains unclear whether CD14 mediates lipopolysaccharide (LPS) signaling in this specialized liver macrophage population. In this series of experiments, we determined the role of CD14 in LPS activation of Kupffer cells by using several complementary approaches. First, we isolated Kupffer cells from human livers and studied the effects of anti-CD14 antibodies on LPS activation of these cells. Kupffer cells were incubated with increasing concentrations of LPS in the presence and absence of recombinant human LPS binding protein (LBP). With increasing concentrations of LPS, human Kupffer cell tumor necrosis factor-α (TNF-α) production (a marker for Kupffer cell activation) increased in a dose-dependent manner in the presence and absence of LBP. In the presence of anti-human CD14 antibodies, the production of TNF-α was significantly diminished. Second, we compared LPS activation of Kupffer cells isolated from wild-type and CD14 knockout mice. Kupffer cells from CD14 knockout mice produced significantly less TNF-α in response to the same amount of LPS. Together, these data strongly support a critical role for CD14 in Kupffer cell responses to LPS.

Using a full via first (FVF) dual damascene (DD) scheme for copper processing with low-k dielectrics has presented many new challenges to the semiconductor industry. Among those challenges, for photolithography, resist poisoning at the trench level has been the most daunting. Resist and bottom anti-reflective coating (BARC) screenings for poisoning at the 0.13 micrometers and 0.10 micrometers technology nodes have been performed on a variety of KrF and ArF resist and BARC platforms using a simple semi-qualitative method. By varying resist parameters such as resin, photoacid generator (PAG), and solvent types, a lithographically suitable KrF resist is found for the 0.13 micrometers node with minimal sensitivity to poisoning. In addition, ArF resists and BARCs were screened for their sensitivity to poisoning for the 0.10 micrometers node. Suitable resist and BARC candidates are identified for preliminary use for the 0.10 micrometers node.

The emergence of multidrug-resistant microbes has serious implications for managing infection and sepsis and has stimulated efforts to develop alternative treatments, such as antimicrobial peptides. The objective of this study was to test a designer peptide, novispirin G10, against multidrug-resistant microorganisms. By two-stage radial diffusion assays, its activity against such organisms compared favorably with that of standard antibiotics and other antimicrobial peptides. It killed bacteria very rapidly, was nonhemolytic, and was relatively noncytotoxic. The peptide induced an immediate, massive efflux of potassium fromPseudomonas aeruginosa, suggesting that it altered the permeability of its inner membrane. The presence of human serum reduced but did not eliminate its activity. We tested the in vivo activity of novispirin G10 in rats with an infected, partial-thickness burn that covered 20% of their total body surface area. The burned area was seeded with 106CFU of a Silvadene-resistantP. aeruginosastrain, and 24 h later a single treatment with 0, 1, 3, or 6 mg of synthetic novispirin G10 (n= 16 at each concentration) per kg was given intradermally. Significant bacterial killing (P< 0.0001) was evident within 4 h in each peptide group compared to controls receiving vehicle. Antimicrobial peptides such as novispirin G10 may provide a useful alternative or adjunct to standard antibiotic agents in treating burns or other wound infections.

Skin is equipped with an array of immune mediators aimed at fighting invading microbes. CD14 has been shown to play a key role in modulating the activation of cells by LPS. Since LPS levels within burn wounds are often found to be elevated, we sought to examine the expression of CD14 within human skin following thermal injury.Patients who sustained partial thickness burns, were recruited into the study (n=57). Total RNA was isolated from both burn and normal (control) skin. Northern blot analysis and TaqMan RT-PCR were used to determine skin CD14 mRNA levels. Immunohistochemistry was used to localize CD14 expression in burned and normal skin.Quantitative PCR showed significantly increased CD14 expression levels in the immediate post-burn period (P0.05 burn versus non-burn). Immunohistochemistry revealed more pronounced CD14 staining 24 h after the injury, reaching normal levels approximately 5-7 days post-burn.CD14 expression peaks within the first week post-burn before declining, reaching normal levels after 14 days. This loss of supranormal CD14 expression locally within the wound may contribute to a weakened host defense response 5-6 days after injury, when patients become especially vulnerable to infection.