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A novel, highly diastereoselective, and metal-free synthesis of multisubstituted piperidines via an S N 1 approach is reported in this study. The method allows for the preparation of highly functionalized compounds with exceptional diastereomeric selectivities and consistently reproducible yields. These compounds are of significant interest due to their remarkable biological activities toward influenza endonuclease.

Metasurfaces have exhibited exceptional proficiency in precisely modulating light properties within narrow wavelength spectra. However, there is a growing demand for multi-resonant metasurfaces capable of wavefront engineering across broad spectral ranges. In this study, we introduce a microcavity-assisted multi-resonant metasurface platform that integrates subwavelength meta-atoms with a specially designed distributed Bragg reflector (DBR) substrate. This platform enables the simultaneous excitation of various resonant modes within the metasurface, resulting in multiple high-Q resonances spanning from the visible to the near-infrared (NIR) regions. The developed metasurface generates up to 15 high-Q resonant peaks across the visible-NIR spectrum, achieving a maximum efficiency of 81% (70.7%) in simulation (experiment) with an average efficiency of 76.6% (54.5%) and a standard deviation of 4.1% (11.1%). Additionally, we demonstrate the versatility of the multi-resonant metasurface in amplitude, phase, and wavefront modulations at peak wavelengths. By integrating structural color printing and vectorial holographic imaging, our proposed metasurface platform shows potential for applications in optical displays and encryption. This work paves the way for the development of next-generation multi-resonant metasurfaces with broad-ranging applications in photonics and beyond., (© 2024. The Author(s).)

Septic arthritis is a frustrating disease in sea turtle rehabilitation because of its unclear pathogenesis, delayed onset during rehabilitation, long-term treatment requirements, and potentially poor prognosis. Radiography, blood cultures, and arthrocentesis have been used as diagnostic tools for suspected cases. However, there is currently a lack of data on the characteristics of synovial fluid in healthy sea turtles. To establish reference data for synovial fluid in sea turtles, we enrolled 14 green turtles Chelonia mydas rescued between 2019 and 2022 from 3 facilities using the following inclusion criteria: normal attitude and appetite, normal motor functions of the 4 limbs, no joint swelling, and no ongoing use of antibiotics for at least 1 mo. Bacterial cultures of blood and synovial fluid from the shoulder joints of these turtles were obtained and a qualitative analysis of the synovial fluid was performed. The results revealed bacterial culture-negative blood and synovial fluids at 37°C. Most characteristics of normal synovial fluid in green turtles, such as being transparent, colorless, and able to create a strand of over 2.5 cm by being pulled with a needle in viscosity trials, as well as the cytology of the normal synovial fluids being dominated by histiocytes and synovial lining cells, lymphocytes, and occasionally a few heterophils or erythrocytes were similar to those in mammals. This study provides information on the normal synovial fluid characteristics of green turtles in Taiwan, which may be beneficial for the diagnosis of joint diseases in sea turtles.

Objective: To integrate the qualitative research on the self-management experience of breast cancer patients and conduct a systematic review of their self-management experience., Methods: Using a computer to search a series of databases such as CNKI, Wanfang, VIP, and China Biomedical Database, systematically collect and integrate qualitative research on the self-management experience of breast cancer patients, and the search time is limited to January 2010 to December 2022. The qualitative research quality evaluation standard of the Joanna Briggs Institute Centre for Evidence-Based Health Care in Australia was used as the evaluation standard of this project to complete the accurate evaluation of the literature; Meta-analysis was used to complete the effective integration of the results., Results: 17 pieces of literature were included in this project, and 37 research results with strong integrity were extracted accordingly. On this basis, 7 different categories were summarised, and three integrated results were obtained: the experience of maintaining self-management, symptom recognition, and self-management., Conclusion: In the different stages of self-management of breast cancer patients, medical staff should give targeted guidance to help patients obtain a good prognosis., (© 2024 The Authors. The International Journal of Health Planning and Management published by John Wiley & Sons Ltd.)

The baculovirus expression system is a powerful and widely used method to generate large quantities of recombinant protein. However, challenges exist in workflows utilizing either liquid baculovirus stocks or the Titerless Infected-Cells Preservation and Scale-Up (TIPS) method, including the time and effort to generate baculoviruses, screen for protein expression and store large numbers of baculovirus stocks. To mitigate these challenges, we have developed a streamlined, hybrid workflow which utilizes high titer liquid virus stocks for rapid plate-based protein expression screening, followed by a TIPS-based scale-up for larger protein production efforts. Additionally, we have automated each step in this screening workflow using a custom robotic system. With these process improvements, we have significantly reduced the time, effort and resources required to manage large baculovirus generation and expression screening campaigns., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Traditional varifocal lenses are bulky and mechanically complex. Emerging active metalenses promise compactness and design flexibility but face issues like mechanical tuning reliability and nonlinear focal length tuning due to additional medium requirements. In this work, we propose a varifocal metalens design based on superimposing light intensity distributions from two orthogonal polarization states. This approach enables continuous and precise focal length control within the visible spectrum, while maintaining relatively high focusing efficiencies (∼41% in simulation and ∼28% in measurement) and quality. In experimental validation, the metalens exhibited flexible tunability, with the focal length continuously adjustable between two spatial positions upon variation of the incident polarization angle. The MTF results showed high contrast reproduction and sharp imaging, with a Strehl ratio of >0.7 for all polarization angles. With compactness, design flexibility, and high focusing quality, the proposed varifocal metalens holds potential for diverse applications, advancing adaptive and versatile optical devices.

Microplastics (MPs) have emerged as a global concern, with a recent study being the first to detect them in the bloodstream of healthy people. However, precise information regarding the toxic effects of MPs on the human vascular system is currently lacking. In this study, we used human vascular endothelial EA. hy926 cells to examine the toxic potential of polystyrene MPs (PSMPs) under realistic blood concentrations. Our findings indicated that PSMPs can cause oxidative stress by reducing the expression of antioxidants, thereby leading to apoptotic cytotoxicity in EA. hy926 cells. Furthermore, the protective potential of heat shock proteins can be reduced by PSMPs. PSMP-induced apoptosis might also lower the expression of rho-associated protein kinase-1 and nuclear factor-κB expression, thus dampening LRR- and pyrin domain-containing protein 3 in EA. hy926 cells. Moreover, we observed that PSMPs induce vascular barrier dysfunction via the depletion of zonula occludens-1 protein. However, although protein expression of the nuclear hormone receptor 77 was inhibited, no significant increase in ectin-like oxidized low-density lipoprotein receptor-1 was noted in PSMP-treated EA. hy926 cells. These results demonstrate that exposure to PSMPs may not sufficiently increase the risk of developing atherosclerosis. Overall, our research signifies that exposure to realistic blood concentrations of PSMPs is associated with low atherosclerotic cardiovascular risk in humans., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier Ltd.)

Tungsten trioxide (WO 3 )-based nanoparticles (NPs) are gaining popularity because of their exciting potential for photocatalytic applications; however, the toxic potential of WO 3 -based NPs remains a concern. In this study, we evaluated the toxic risk of WO 3 NPs and hydrated WO 3 NPs (WO 3 ·H 2 O NPs) using lung cells and explored the underlying mechanism. WO 3 NPs and WO 3 ·H 2 O NPs significantly decreased the number of viable cells (59.5 %-85.8 % of control) and promoted apoptosis in human alveolar basal epithelial A549 cells after a 24-h exposure. Both WO 3 NPs and WO 3 ·H 2 O NPs reduced the expression of heme oxygenase-1 (0.15-0.33 folds of control) and superoxide dismutase 2 (0.31-0.66 folds of control) and increased reactive oxygen species production (1.4-2.6 folds of control) and 8-hydroxy-2'-deoxyguanosine accumulation (1.22-1.43 folds of control). The results showed that WO 3 NPs have higher cytotoxicity and oxidative potential than WO 3 ·H 2 O NPs. In addition, the WO 3 NP cellular uptake rate was significantly higher than the WO 3 ·H 2 O NPs uptake rate in pulmonary cells. The greater extent of oxidative adverse effects induced by WO 3 -based NPs appears to be related to the enhanced particle uptake. WO 3 NPs and WO 3 ·H 2 O NPs exposure led to the secretion of inflammatory factor interleukin 6 (1.63-3.42 folds of control). Decreases in serpin family A member 1 gene expression (0.28-0.58 folds of control) and increases in the oxidation of neutrophil elastase inhibitor (1.34-1.62 folds of control) in pulmonary cells also suggest that exposure to WO 3 NPs and WO 3 ·H 2 O NPs raises the risk of developing chronic obstructive pulmonary disease. Taken together, our findings indicate that the toxic risk of WO 3 NPs and WO 3 ·H 2 O NPs must be considered when manufacturing and applying WO 3 -based NPs., Competing Interests: Declaration of competing interest All the authors declare no conflict of interest., (Copyright © 2022. Published by Elsevier B.V.)

Human metapneumovirus (hMPV) belongs to the Pneumoviridae family and is closely related to respiratory syncytial virus (RSV). The surface fusion (F) glycoprotein mediates viral fusion and is the primary target of neutralizing antibodies against hMPV. Here we report 113 hMPV-F specific monoclonal antibodies (mAbs) isolated from memory B cells of human donors. We characterize the antibodies' germline usage, epitopes, neutralization potencies, and binding specificities. We find that unlike RSV-F specific mAbs, antibody responses to hMPV F are less dominant against the apex of the antigen, and the majority of the potent neutralizing mAbs recognize epitopes on the side of hMPV F. Furthermore, neutralizing epitopes that differ from previously defined antigenic sites on RSV F are identified, and multiple binding modes of site V and II mAbs are discovered. Interestingly, mAbs that bind preferentially to the unprocessed prefusion F show poor neutralization potency. These results elucidate the immune recognition of hMPV infection and provide novel insights for future hMPV antibody and vaccine development., (© 2022. Merck & Co., Inc., Kenilworth, NJ, USA and its affiliates.)

In this work, a cost-effective wax was synthesized from waste cooking oil (WCO), and its properties including melting point, color, hardness, combustion performance and micro-morphology were tested and analyzed. The obtained results showed that the epoxy waste cooking oil had lighter color, higher melting point and hardness than that of original WCO, which could be used as wax. Moreover, introducing stearic acid further improved the performances of WCO-based wax. The WCO-based wax made of epoxy waste cooking oil and stearic acid (containing ≥50 wt% stearic acid) displayed a relatively high melting point (≥46 °C), light color (Lovibond color code Y ≤ 16.1, R ≤ 2.3), good hardness (needle penetration index ≤2.95 mm) and long combustion time (≥227 min), and could achieve the required national standard and be used as a substitute for the commercially available soybean wax. Together with many additional benefits such as low synthesis cost, mild reaction conditions, convenient synthesis route, and no secondary pollution, producing wax based on WCO could provide a new path for WCO recycling in economically trailing regions., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

TREM2 has been identified by genomic analysis as a potential and novel target for the treatment of Alzheimer's disease. To enable structure-based screening of potential small molecule therapeutics, we sought to develop a robust crystallization platform for the TREM2 Ig-like domain. A systematic set of constructs containing the structural chaperone, maltose binding protein (MBP), fused to the Ig domain of TREM2, were evaluated in parallel expression and purification, followed by crystallization studies. Using protein crystallization and high-resolution diffraction as a readout, a MBP-TREM2 Ig fusion construct was identified that generates reproducible protein crystals diffracting at 2.0 Å, which makes it suitable for soaking of potential ligands. Importantly, analysis of crystal packing interfaces indicates that most of the surface of the TREM2 Ig domain is available for small molecule binding. A proof of concept co-crystallization study with a small library of fragments validated potential utility of this system for the discovery of new TREM2 therapeutics., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Respiratory syncytial virus (RSV) infection is the leading cause of hospitalization and infant mortality under six months of age worldwide; therefore, the prevention of RSV infection in all infants represents a significant unmet medical need. Here we report the isolation of a potent and broadly neutralizing RSV monoclonal antibody derived from a human memory B-cell. This antibody, RB1, is equipotent on RSV A and B subtypes, potently neutralizes a diverse panel of clinical isolates in vitro and demonstrates in vivo protection. It binds to a highly conserved epitope in antigenic site IV of the RSV fusion glycoprotein. RB1 is the parental antibody to MK-1654 which is currently in clinical development for the prevention of RSV infection in infants.

Background: We examined the risk for Group B streptococcus (GBS)-related diseases in newborns born to mothers who participated in a universal GBS screening program and to determine whether differences are observed in factors affecting the morbidity for neonatal early-onset GBS-related diseases., Methods: This is a retrospective study and the study subjects were women who had undergone GBS screening and who gave birth naturally and their newborns between April 15, 2012 and December 31, 2013. Data from the GBS screening system database and the National Health Insurance database were collected to calculate the GBS prevalence in pregnant women and morbidity of newborns with early-onset GBS-related diseases., Results: The GBS prevalence in pregnant women who gave birth naturally was 19.58%. The rate of early-onset infection caused by GBS in newborns decreased from the original 0.1% to 0.02%, a decrease of as high as 80%. After the implementation of the universal GBS screening program, only three factors, including positive GBS screening result (OR = 2.84), CCI (OR = 2.45), and preterm birth (OR = 4.81) affected the morbidity for neonatal early-onset GBS-related diseases, whereas other factors had no significant impact., Conclusion: The implementation of the universal GBS screening program decreased the infection rate of neonatal early-onset GBS diseases. The effects of socioeconomic factors and high-risk pregnancy on early-onset GBS infections were weakened.

An internal HTS effort identified a novel PDE2 inhibitor series that was subsequently optimized for improved PDE2 activity and off-target selectivity. The optimized lead, compound 4, improved cognitive performance in a rodent novel object recognition task as well as a non-human primate object retrieval task. In addition, co-crystallization studies of close analog of 4 in the PDE2 active site revealed unique binding interactions influencing the high PDE isoform selectivity., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Objectives: The increasing prevalence of mutations in HIV-1 reverse transcriptase (RT) that confer resistance to existing NRTIs and NNRTIs underscores the need to develop RT inhibitors with novel mode-of-inhibition and distinct resistance profiles., Methods: Biochemical assays were employed to identify inhibitors of RT activity and characterize their mode of inhibition. The antiviral activity of the inhibitors was assessed by cell-based assays using laboratory HIV-1 isolates and MT4 cells. RT variants were purified via avidin affinity columns., Results: Compound A displayed equal or greater potency against many common NNRTI-resistant RTs (K103N and Y181C RTs) relative to WT RT. Despite possessing certain NNRTI-like properties, such as being unable to inhibit an engineered variant of RT lacking an NNRTI-binding pocket, we found that compound A was dependent on Mg2+ for binding to RT. Optimization of compound A led to more potent analogues, which retained similar activities against WT and K103N mutant viruses with submicromolar potency in a cell-based assay. One of the analogues, compound G, was crystallized in complex with RT and the structure was determined at 2.6 Å resolution. The structure indicated that compound G simultaneously interacts with the active site (Asp186), the highly conserved primer grip region (Leu234 and Trp229) and the NNRTI-binding pocket (Tyr188)., Conclusions: These findings reveal a novel class of RT bifunctional inhibitors that are not sensitive to the most common RT mutations, which can be further developed to address the deficiency of current RT inhibitors., (© The Author 2017. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

We have identified a novel PDE2 inhibitor series using fragment-based screening. Pyrazolopyrimidine fragment 1, while possessing weak potency (K i  = 22.4 μM), exhibited good binding efficiencies (LBE = 0.49, LLE = 4.48) to serve as a start for structure-based drug design. With the assistance of molecular modeling and X-ray crystallography, this fragment was developed into a series of potent PDE2 inhibitors with good physicochemical properties. Compound 16, a PDE2 selective inhibitor, was identified that exhibited favorable rat pharmacokinetic properties., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Using the HIV-1 protease binding mode of MK-8718 and PL-100 as inspiration, a novel aspartate binding bicyclic piperazine sulfonamide core was designed and synthesized. The resulting HIV-1 protease inhibitor containing this core showed an 60-fold increase in enzyme binding affinity and a 10-fold increase in antiviral activity relative to MK-8718 ., Competing Interests: The authors declare no competing financial interest.

Current therapies for chronic pain can have insufficient efficacy and lead to side effects, necessitating research of novel targets against pain. Although originally identified as an oncogene, Tropomyosin-related kinase A (TrkA) is linked to pain and elevated levels of NGF (the ligand for TrkA) are associated with chronic pain. Antibodies that block TrkA interaction with its ligand, NGF, are in clinical trials for pain relief. Here, we describe the identification of TrkA-specific inhibitors and the structural basis for their selectivity over other Trk family kinases. The X-ray structures reveal a binding site outside the kinase active site that uses residues from the kinase domain and the juxtamembrane region. Three modes of binding with the juxtamembrane region are characterized through a series of ligand-bound complexes. The structures indicate a critical pharmacophore on the compounds that leads to the distinct binding modes. The mode of interaction can allow TrkA selectivity over TrkB and TrkC or promiscuous, pan-Trk inhibition. This finding highlights the difficulty in characterizing the structure-activity relationship of a chemical series in the absence of structural information because of substantial differences in the interacting residues. These structures illustrate the flexibility of binding to sequences outside of-but adjacent to-the kinase domain of TrkA. This knowledge allows development of compounds with specificity for TrkA or the family of Trk proteins., Competing Interests: The work described in this manuscript was performed while all authors were employed at Merck and Co., Inc.

The initial structure activity relationships around an isoindoline uHTS hit will be described. Information gleaned from ligand co-crystal structures allowed for rapid refinements in both MARK potency and kinase selectivity. These efforts allowed for the identification of a compound with properties suitable for use as an in vitro tool compound for validation studies on MARK as a viable target for Alzheimer's disease., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Inhibition of microtubule affinity regulating kinase (MARK) represents a potentially attractive means of arresting neurofibrillary tangle pathology in Alzheimer's disease. This manuscript outlines efforts to optimize a pyrazolopyrimidine series of MARK inhibitors by focusing on improvements in potency, physical properties and attributes amenable to CNS penetration. A unique cylcyclohexyldiamine scaffold was identified that led to remarkable improvements in potency, opening up opportunities to reduce MW, Pgp efflux and improve pharmacokinetic properties while also conferring improved solubility., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: The Taiwanese government considers fluoride varnish to be a major component of preventive dental cares for preschool children. This study aimed to explore the extent of utilization of fluoride varnish and its determining factors among Taiwanese preschool children., Methods: Using preschool children under the age of 5 years as our participants, this study was conducted based on the 2008 Taiwan database of the Ministry of the Interior, linked with information gathered between 2006 and 2008 on preventive healthcare and health insurance from the Bureau of Health Promotion and the National Health Research Institute. A total of 949,023 preschool children (< 5 years old) were identified to meet the requirement for fluoride varnish services., Results: The percentage of Taiwanese preschool children that used fluoride varnish was 34.85%.The probability that fluoride varnish would be utilized was higher among children with catastrophic illness/injury or relevant chronic illnesses than those without. In addition, the probability of children with disabilities using fluoride varnish was lower than that of nondisabled children. Finally, parent sex, parent age, urbanization level of residence, and parents' premium-based salary significantly affected the children's utilization probability of fluoride varnish., Conclusion: In order to increase the utilization of fluoride varnish among preschool children in Taiwan in the future, target groups consisting of females, children < 3 years of age, and disabled children should be prioritized. Parental factors are also important in affecting the utilization of fluoride varnish in children., (Copyright © 2016. Published by Elsevier Taiwan LLC.)

A novel HIV protease inhibitor was designed using a morpholine core as the aspartate binding group. Analysis of the crystal structure of the initial lead bound to HIV protease enabled optimization of enzyme potency and antiviral activity. This afforded a series of potent orally bioavailable inhibitors of which MK-8718 was identified as a compound with a favorable overall profile.

Unlabelled: The highly conserved H3 poxvirus protein is a major target of the human antibody response against poxviruses and is likely a key contributor to protection against infection. Here, we present the crystal structure of H3 from vaccinia virus at a 1.9-Å resolution. H3 looks like a glycosyltransferase, a family of enzymes that transfer carbohydrate molecules to a variety of acceptor substrates. Like glycosyltransferases, H3 binds UDP-glucose, as shown by saturation transfer difference (STD) nuclear magnetic resonance (NMR) spectroscopy, and this binding requires Mg(2+) Mutation of the glycosyltransferase-like metal ion binding motif in H3 greatly diminished its binding to UDP-glucose. We found by flow cytometry that H3 binds to the surface of human cells but does not bind well to cells that are deficient in surface glycosaminoglycans. STD NMR experiments using a heparin sulfate decasaccharide confirmed that H3 binds heparin sulfate. We propose that a surface of H3 with an excess positive charge may be the binding site for heparin. Heparin binding and glycosyltransferase activity may be involved in the function of H3 in the poxvirus life cycle., Importance: Poxviruses are under intense research because of bioterrorism concerns, zoonotic infections, and the side effects of existing smallpox vaccines. The smallpox vaccine using vaccinia virus has been highly successful, but it is still unclear why the vaccine is so effective. Studying the antigens that the immune system recognizes may allow a better understanding of how the vaccine elicits immunity and how improved vaccines can be developed. Poxvirus protein H3 is a major target of the immune system. The H3 crystal structure shows that it has a glycosyltransferase protein fold. We demonstrate that H3 binds the sugar nucleotide UDP-glucose, as do glycosyltransferases. Our experiments also reveal that H3 binds cell surface molecules that are involved in the attachment of poxviruses to cells. These structural and functional studies of H3 will help in designing better vaccines and therapeutics., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

Background: The oral condition of people with disabilities has considerable influence on their physical and mental health. However, nationwide surveys regarding this group have not been conducted. For this study, we used the National Health Insurance Research Database to explore the tooth filling utilization among people with disabilities., Methods: Using the database of the Ministry of the Interior in 2008 which included people with disabilities registered, we merged with the medical claims database in 2008 of the Bureau of National Health Insurance to calculate the tooth filling utilization and to analyze relative factors. We recruited 993,487 people with disabilities as the research sample., Results: The tooth filling utilization was 17.53 %. The multiple logistic regression result showed that the utilization rate of men was lower than that of women (OR = 0.78, 95 % CI = 0.77-0.79) and older people had lower utilization rates (aged over 75, OR = 0.22, 95 % CI = 0.22-0.23) compared to those under the age of 20. Other factors that significantly influenced the low tooth filling utilization included a low education level, living in less urbanized areas, low economic capacity, dementia, and severe disability., Conclusion: We identified the factors that influence and decrease the tooth-filling service utilization rate: male sex, old age, low education level, being married, indigenous ethnicity, residing in a low urbanization area, low income, chronic circulatory system diseases, dementia, and severe disabilities. We suggest establishing proper medical care environments for high-risk groups to maintain their quality of life.

Objective: To examine the variations in the prevalence of skin reactions and the association between skin reactions and efficacy of summer acupoint application treatment (SAAT) on chronic pulmonary disease (CPD)., Methods: A total of 2,038 patients with CPD were enrolled at 3 independent hospitals (defined as Groups A, B and C, respectively) in China. All patients were treated by SAAT, as applying a herbal paste onto the acupoints of Fengmen (BL 12) and Feishu (BL 13) on the dog days of summer, according to the lunar calendar, in 2008. Ten days after treatment, skin reaction data (no reaction, itching, stinging, blistering, and infection) were obtained via face-to-face interviews. Patients were retreated in the same hospital one year later, thereby allowing doctors to assess treatment efficacy based on the patients' symptoms, the severity of the spirometric abnormalities, and the concomitant medications used., Results: A large number of patients (85.3%) displayed reactive symptoms; however, the marked associations between reactive symptoms and age or gender were not observed. An increased number of patients from Group B (99.3%) and Group C (76.5%) displayed reactive symptoms due to the increased mass of crude Semen Sinapis Albae. The effective rate of SAAT was as high as 90.4% for patients of Group B, which was followed by Group A (70.9%) and Group C (42.2%). Using stratified analyses, a convincing association between reactive symptoms and therapeutic efficacy was observed for patients with asthma [itching: odds ratio (OR)=2.17, 95% confidence interval (CI): 1.49 to 3.14; blistering: OR=0.43, 95% CI: 0.25 to 0.73; and no reaction: OR=0.56, 95% CI: 0.35 to 0.90]. However, the same tendency was not observed for patients with chronic bronchitis and chronic obstructive pulmonary disease., Conclusions: SAAT can induce very mild skin reactions for patients with CPD, among which patients with asthma displayed a strong association between skin reactions and therapeutic efficacy. The skin reactions may be induced by the crude Semen Sinapis Albae.

We demonstrated a simple and cost-effective method to fabricate all fiber Mach-Zehnder interferometer (MZI) based on cascading a short section of liquid crystal (LC)-filled hollow-optic fiber (HOF) between two single mode fibers by using automatically splicing technique. The transmission spectra of the proposed MZI with different LC-infiltrated length were measured and the temperature-induced wavelength shifts of the interference fringes were recorded. Both blue shift and red shift were observed, depending the temperature range. Based on our experimental results, interference fringe was observed with a maximum interferometric contrast over 35dB. The temperature-induced resonant wavelength blue-shifts 70.4 nm for the MZI with an LC length of 9.79 mm and the wavelength temperature sensitivity of -1.55 nm/°C is easily achieved as the temperature increases from 25°C to 77°C.

Periodontitis and osteoporosis are primary concerns in public health and clinical management. The aim of this study was to investigate the association between periodontitis and osteoporosis by gender.Data were retrieved from the National Health Insurance Research Database, Taiwan. A diagnosis of periodontitis was defined on the basis of subgingival curettage, periodontal flap operation, and gingivectomy (excluding those with restorative or aesthetic indications). Multiple logistic regression was used for analysis. After adjusting for age, sex, income, and geographical region, there was a significant association between periodontitis and osteoporosis among women (odds ratio: 1.96; 95% confidence interval 1.17-3.26). The association between periodontitis and osteoporosis was significant among women.

Limited studies with large samples have been conducted on the utilization of dental calculus scaling among people with physical or mental disabilities. This study aimed to investigate the utilization of dental calculus scaling among the national disabled population. This study analyzed the utilization of dental calculus scaling among the disabled people, using the nationwide data between 2006 and 2008. Descriptive analysis and logistic regression were performed to analyze related influential factors for dental calculus scaling utilization. The dental calculus scaling utilization rate among people with physical or mental disabilities was 16.39%, and the annual utilization frequency was 0.2 times. Utilization rate was higher among the female and non-aboriginal samples. Utilization rate decreased with increased age and disability severity while utilization rate increased with income, education level, urbanization of residential area and number of chronic illnesses. Related influential factors for dental calculus scaling utilization rate were gender, age, ethnicity (aboriginal or non-aboriginal), education level, urbanization of residence area, income, catastrophic illnesses, chronic illnesses, disability types, and disability severity significantly influenced the dental calculus scaling utilization rate., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

We have identified several series of small molecule inhibitors of TrkA with unique binding modes. The starting leads were chosen to maximize the structural and binding mode diversity derived from a high throughput screen of our internal compound collection. These leads were optimized for potency and selectivity employing a structure based drug design approach adhering to the principles of ligand efficiency to maximize binding affinity without overly relying on lipophilic interactions. This endeavor resulted in the identification of several small molecule pan-Trk inhibitor series that exhibit high selectivity for TrkA/B/C versus a diverse panel of kinases. We have also demonstrated efficacy in both inflammatory and neuropathic pain models upon oral dosing. Herein we describe the identification process, hit-to-lead progression, and binding profiles of these selective pan-Trk kinase inhibitors.

Efficient oral endotracheal intubation of laboratory animals is a challenging technique in veterinary research. This study introduces a miniaturized lighted stylet for rabbit intubation. An experiment with repeated measures on two factors was used to assess the feasibility and efficacy of this method. The first factor compared stylet intubation vs. laryngoscopic intubation. The second compared three practitioners, one with prior experience and two without. Success rates on the initial attempt were not statistically different (χ(2)=2.46, P=0.12). The time difference between methods was significant (F=41.007, P<0.001), although the effect of practitioners was not (F=1.038, P=0.365). The mean±SD of the intubation time, combining results from the three practitioners, was 20.34±17.15s for the stylet method and 57.58±64.21s for the laryngoscopic method. The results of this study demonstrate that lighted stylet intubation is efficient, robust, and independent of practitioner experience., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The JAK-STAT pathway mediates signaling by cytokines, which control survival, proliferation, and differentiation of a variety of cells. In recent years, a single point mutation (V617F) in the tyrosine kinase JAK2 was found to be present with a high incidence in myeloproliferative disorders (MPDs). This mutation led to hyperactivation of JAK2, cytokine-independent signaling, and subsequent activation of downstream signaling networks. The genetic, biological, and physiological evidence suggests that JAK2 inhibitors could be effective in treating MPDs. De novo design efforts of new scaffolds identified 1-amino-5H-pyrido[4,3-b]indol-4-carboxamides as a new viable lead series. Subsequent optimization of cell potency, metabolic stability, and off-target activities of the leads led to the discovery of 7-(2-aminopyrimidin-5-yl)-1-{[(1R)-1-cyclopropyl-2,2,2-trifluoroethyl]amino}-5H-pyrido[4,3-b]indole-4-carboxamide (65). Compound 65 is a potent, orally active inhibitor of JAK2 with excellent selectivity, PK profile, and in vivo efficacy in animal models.

HIV/AIDS continues to be a menace to public health. Several drugs currently on the market have successfully improved the ability to manage the viral burden in infected patients. However, new drugs are needed to combat the rapid emergence of mutated forms of the virus that are resistant to existing therapies. Currently, approved drugs target three of the four major enzyme activities encoded by the virus that are critical to the HIV life cycle. Although a number of inhibitors of HIV RNase H activity have been reported, few inhibit by directly engaging the RNase H active site. Here, we describe structures of naphthyridinone-containing inhibitors bound to the RNase H active site. This class of compounds binds to the active site via two metal ions that are coordinated by catalytic site residues, D443, E478, D498, and D549. The directionality of the naphthyridinone pharmacophore is restricted by the ordering of D549 and H539 in the RNase H domain. In addition, one of the naphthyridinone-based compounds was found to bind at a second site close to the polymerase active site and non-nucleoside/nucleotide inhibitor sites in a metal-independent manner. Further characterization, using fluorescence-based thermal denaturation and a crystal structure of the isolated RNase H domain reveals that this compound can also bind the RNase H site and retains the metal-dependent binding mode of this class of molecules. These structures provide a means for structurally guided design of novel RNase H inhibitors.

A series of HIV-1 protease inhibitors containing an epsilon substituted lysinol backbone was synthesized. Two novel synthetic routes using N-boc-L-glutamic acid alpha-benzyl ester and 2,6-diaminopimelic acid were developed. Incorporation of this epsilon substituent enabled access to the S2 pocket of the enzyme, affording high potency inhibitors. Modeling studies and synthetic efforts suggest the potency increase is due to both conformational bias and van der Waals interactions with the S2 pocket., (2010 Elsevier Ltd. All rights reserved.)

Objective: To investigate the expressions of 15- and 5-lipoxygenases in leukocytes and the changes of the levels of blood lipoxin A₄ (LXA₄) and leukotriene C₄ (LTC₄) in children with asthma., Methods: The mRNA levels of 15- and 5-lipoxygenases in leukocytes were assessed by RT-PCR, and the levels of blood LXA₄ and LTC₄ were determined by ELISA, in 106 children with mild, moderate and severe asthma. Forty healthy children served as the controls., Results: In children with mild, moderate and severe asthma, the relative mRNA levels of 15-lipoxygenase in leukocytes were 1.78 ± 0.56, 1.28 ± 0.45 and 0.58 ± 0.22 (F = 16.72, P < 0.01), respectively, and all were higher than that of the controls (0.26 ± 0.12, P < 0.05). The levels of blood LXA₄ were (5.52 ± 1.97), (1.86 ± 0.72) and (0.81 ± 0.36) µg/L (F = 22.59, P < 0.01), respectively, decreasing with the severity of asthma, and all were higher than that of the controls [(0.04 ± 0.01) µg/L, P < 0.05]. There was a positive correlation between PEF, FEV(1) and blood LXA₄. The relative levels of 5-lipoxygenase mRNA in leukocytes were 0.26 ± 0.12, 0.79 ± 0.34 and 1.21 ± 0.52, respectively in children with asthma of mild, moderate and severe degree (F = 18.64, P < 0.01), which showed an increase with the severity of the disease, and all of which were higher than that of the controls (0.12 ± 0.05, P < 0.05). The levels of blood LTC₄ were (22.4 ± 8.2), (54.6 ± 28.4) and (118.7 ± 41.1) ng/L (F = 25.91, P < 0.01), respectively, also showing an increase with the severity of asthma, and were higher than that of the controls [(6.8 ± 2.5) ng/L, P < 0.05]. There was a negative correlation between PEF, FEV₁ and blood LTC₄., Conclusion: The reversed changes of 15-lipoxygenase product LXA₄ and 5-lipoxygenase product LTC₄ in children with asthma of mild, moderate and severe degree suggests that insufficiency of LXA₄, an physiological antagonist to leukotrienes, and an overproduction of LTC₄, may be involved in the pathogenesis of worsening of asthma in children.

Background: The objective of this study was to develop a low-fat (LF) mayonnaise containing polysaccharide gums as functional ingredients. Xanthan gum (XG, 15 g kg(-1)), citrus fiber (CF, 100 g kg(-1)) and variable concentration of guar gum (GG) were used to formulate the optimum ratios of polysaccharide gums as fat replacers. The fat content in LF mayonnaise was reduced to 50% if compared with full-fat (FF) mayonnaise, and the products still maintained ideal rheological properties., Results: The rheological parameters showed that there were no (P > 0.05) differences in yield stress, viscosity and flow behavior index between XG + 10 g kg(-1) GG, CF + 5 g kg(-1) GG and FF control. LF mayonnaises had lower caloric values and higher dietary fiber content than the FF counterpart. Scanning electron microscopy (SEM) micrographs illustrated that the network of aggregated droplets in LF treatments contained a large number of interspaced voids of varying dimensions. Furthermore, in a comparison of sensory evaluation of LF treatments with commercial and our FF mayonnaises, there were no (P > 0.05) differences in any sensory scores among XG + 10 g kg(-1) GG control., Conclusion: This study shows that XG + 10 g kg(-1) GG and CF + 5 g kg(-1) GG could be used in LF mayonnaise formulations based on its multiple functions on processing properties., ((c) 2010 Society of Chemical Industry.)

The current vaccine against smallpox is an infectious form of vaccinia virus that has significant side effects. Alternative vaccine approaches using recombinant viral proteins are being developed. A target of subunit vaccine strategies is the poxvirus protein A33, a conserved protein in the Chordopoxvirinae subfamily of Poxviridae that is expressed on the outer viral envelope. Here we have determined the structure of the A33 ectodomain of vaccinia virus. The structure revealed C-type lectin-like domains (CTLDs) that occur as dimers in A33 crystals with five different crystal lattices. Comparison of the A33 dimer models shows that the A33 monomers have a degree of flexibility in position within the dimer. Structural comparisons show that the A33 monomer is a close match to the Link module class of CTLDs but that the A33 dimer is most similar to the natural killer (NK)-cell receptor class of CTLDs. Structural data on Link modules and NK-cell receptor-ligand complexes suggest a surface of A33 that could interact with viral or host ligands. The dimer interface is well conserved in all known A33 sequences, indicating an important role for the A33 dimer. The structure indicates how previously described A33 mutations disrupt protein folding and locates the positions of N-linked glycosylations and the epitope of a protective antibody.

The genus Legionella contains a diverse group of motile, asaccharolytic, nutritionally fastidious gram-negative rods. Legionella pneumophila is the most important human pathogen, followed by L. micdadei, L. longbeachae, L. dumoffii, and other rare species. Accurate identification of Legionella spp. other than L. pneumophila is difficult because of biochemical inertness and phenotypic identity of different species. The feasibility of using an oligonucleotide array for identification of 18 species of Legionella was evaluated in this study. The method consisted of PCR amplification of the macrophage infectivity potentiator mip gene, followed by hybridization of the digoxigenin-labeled PCR products to a panel of 30 oligonucleotide probes (16- to 24-mers) immobilized on a nylon membrane. A collection of 144 target strains (strains we aimed to identify) and 50 nontarget strains (44 species) were analyzed by the array. Both test sensitivity (144/144 strains) and specificity (50/50 strains) of the array were 100%. The whole procedure for identification of Legionella species by the array can be finished within a working day, starting from isolated colonies. It was concluded that species identification of clinically relevant Legionella spp. by the array method is very reliable and can be used as an accurate alternative to conventional or other molecular methods for identification of Legionella spp.

Prostasin (also called channel activating protease-1 (CAP1)) is an extracellular serine protease implicated in the modulation of fluid and electrolyte regulation via proteolysis of the epithelial sodium channel. Several disease states, particularly hypertension, can be affected by modulation of epithelial sodium channel activity. Thus, understanding the biochemical function of prostasin and developing specific agents to inhibit its activity could have a significant impact on a widespread disease. We report the expression of the prostasin proenzyme in Escherichia coli as insoluble inclusion bodies, refolding and activating via proteolytic removal of the N-terminal propeptide. The refolded and activated enzyme was shown to be pure and monomeric, with kinetic characteristics very similar to prostasin expressed from eukaryotic systems. Active prostasin was crystallized, and the structure was determined to 1.45 A resolution. These apoprotein crystals were soaked with nafamostat, allowing the structure of the inhibited acyl-enzyme intermediate structure to be determined to 2.0 A resolution. Comparison of the inhibited and apoprotein forms of prostasin suggest a mechanism of regulation through stabilization of a loop which interferes with substrate recognition.

Plasmodium falciparum malaria parasites, living in red blood cells, express proteins of the erythrocyte membrane protein-1 (PfEMP1) family on the red blood cell surface. The binding of PfEMP1 molecules to human cell surface receptors mediates the adherence of infected red blood cells to human tissues. The sequences of the 60 PfEMP1 genes in each parasite genome vary greatly from parasite to parasite, yet the variant PfEMP1 proteins maintain receptor binding. Almost all parasites isolated directly from patients bind the human CD36 receptor. Of the several kinds of highly polymorphic cysteine-rich interdomain region (CIDR) domains classified by sequence, only the CIDR1alpha domains bind CD36. Here we describe the CD36-binding portion of a CIDR1alpha domain, MC179, as a bundle of three alpha-helices that are connected by a loop and three additional helices. The MC179 structure, containing seven conserved cysteines and 10 conserved hydrophobic residues, predicts similar structures for the hundreds of CIDR sequences from the many genome sequences now known. Comparison of MC179 with the CIDR domains in the genome of the P. falciparum 3D7 strain provides insights into CIDR domain structure. The CIDR1alpha three-helix bundle exhibits less than 20% sequence identity with the three-helix bundles of Duffy-binding like (DBL) domains, but the two kinds of bundles are almost identical. Despite the enormous diversity of PfEMP1 sequences, the CIDR1alpha and DBL protein structures, taken together, predict that a PfEMP1 molecule is a polymer of three-helix bundles elaborated by a variety of connecting helices and loops. From the structures also comes the insight that DBL1alpha domains are approximately 100 residues larger and that CIDR1alpha domains are approximately 100 residues smaller than sequence alignments predict. This new understanding of PfEMP1 structure will allow the use of better-defined PfEMP1 domains for functional studies, for the design of candidate vaccines, and for understanding the molecular basis of cytoadherence.

Antibody responses are critical components of protective immune responses to many pathogens, but parameters determining which proteins are targeted remain unclear. Vaccination with individual MHC-II-restricted vaccinia virus (VACV, smallpox vaccine) epitopes revealed that CD4(+) T cell help to B cells was surprisingly nontransferable to other virion protein specificities. Many VACV CD4(+) T cell responses identified in an unbiased screen targeted antibody virion protein targets, consistent with deterministic linkage between specificities. We tested the deterministic linkage model by efficiently predicting new vaccinia MHC II epitopes (830% improved efficiency). Finally, we showed CD4(+) T cell help was limiting for neutralizing antibody development and protective immunity in vivo. In contrast to the standard model, these data indicate individual proteins are the unit of B cell-T cell recognition for a large virus. Therefore, MHC restriction is a key selective event for the antiviral antibody response and is probably important for vaccine development to large pathogens.

The smallpox vaccine is widely considered the gold standard for human vaccines, yet the key antibody targets in humans remain unclear. We endeavored to identify a stereotypic, dominant, mature virion (MV) neutralizing antibody target in humans which could be used as a diagnostic serological marker of protective humoral immunity induced by the smallpox vaccine (vaccinia virus [VACV]). We have instead found that diversity is a defining characteristic of the human antibody response to the smallpox vaccine. We show that H3 is the most immunodominant VACV neutralizing antibody target, as determined by correlation analysis of immunoglobulin G (IgG) specificities to MV neutralizing antibody titers. It was determined that purified human anti-H3 IgG is sufficient for neutralization of VACV; however, depletion or blockade of anti-H3 antibodies revealed no significant reduction in neutralization activity, showing anti-H3 IgG is not required in vaccinated humans (or mice) for neutralization of MV. Comparable results were obtained for human (and mouse) anti-L1 IgG and even for anti-H3 and anti-L1 IgG in combination. In addition to H3 and L1, human antibody responses to D8, A27, D13, and A14 exhibited statistically significant correlations with virus neutralization. Altogether, these data indicate the smallpox vaccine succeeds in generating strong neutralizing antibody responses not by eliciting a stereotypic response to a single key antigen but instead by driving development of neutralizing antibodies to multiple viral proteins, resulting in a "safety net" of highly redundant neutralizing antibody responses, the specificities of which can vary from individual to individual. We propose that this is a fundamental attribute of the smallpox vaccine.

Signaling through the programmed death 1 (PD-1) inhibitory receptor upon binding its ligand, PD-L1, suppresses immune responses against autoantigens and tumors and plays an important role in the maintenance of peripheral immune tolerance. Release from PD-1 inhibitory signaling revives "exhausted" virus-specific T cells in chronic viral infections. Here we present the crystal structure of murine PD-1 in complex with human PD-L1. PD-1 and PD-L1 interact through the conserved front and side of their Ig variable (IgV) domains, as do the IgV domains of antibodies and T cell receptors. This places the loops at the ends of the IgV domains on the same side of the PD-1/PD-L1 complex, forming a surface that is similar to the antigen-binding surface of antibodies and T cell receptors. Mapping conserved residues allowed the identification of residues that are important in forming the PD-1/PD-L1 interface. Based on the structure, we show that some reported loss-of-binding mutations involve the PD-1/PD-L1 interaction but that others compromise protein folding. The PD-1/PD-L1 interaction described here may be blocked by antibodies or by designed small-molecule drugs to lower inhibitory signaling that results in a stronger immune response. The immune receptor-like loops offer a new surface for further study and potentially the design of molecules that would affect PD-1/PD-L1 complex formation and thereby modulate the immune response.

This paper proposes a two-stage approach to positioning and identifying tracheal intubations by computer-assisted diagnosis system. First, an innovated LED-based lightwand is developed for positioning the opening of the trachea and inserting the endotracheal tube rapidly. The proposed LED-based lightwand instrument significantly reduces the effects of temperature reaction without changing transillumination through the tissue. After intubation, breath sound analysis instrumentation is developed and assists for identifying the accurate tube position. To overcome the fast and overstepped variations of amplitude, a high sensitivity omni dimensional microphone and an automatic gain control device with linear phase property are calibrated and conducted into experiments of operating intubations. User-friendly interface software is also developed to analyze and transcribe the physiological characteristics of the collected breath sound corpus. Several subjective and objective experiments are performed to examine the practicability of the proposed approach and systems. The preliminary results show that the proposed LED-based lightwand instrument outperformed in the tip temperature without the influence on transillumination. The proposed parameters of average vibration parameter variance (AVPV) and frequency-domain energy variance (FEV) revealed the potential for distinguishing between the tracheal and esophageal intubations.

Medical countermeasures to prevent or treat smallpox are needed due to the potential use of poxviruses as biological weapons. Safety concerns with the currently available smallpox vaccine indicate a need for research on alternative poxvirus vaccine strategies. Molecular vaccines involving the use of proteins and/or genes and recombinant antibodies are among the strategies under current investigation. The poxvirus L1 protein, encoded by the L1R open reading frame, is the target of neutralizing antibodies and has been successfully used as a component of both protein subunit and DNA vaccines. L1-specific monoclonal antibodies (e.g., mouse monoclonal antibody mAb-7D11, mAb-10F5) with potent neutralizing activity bind L1 in a conformation-specific manner. This suggests that proper folding of the L1 protein used in molecular vaccines will affect the production of neutralizing antibodies and protection. Here, we co-crystallized the Fab fragment of mAb-7D11 with the L1 protein. The crystal structure of the complex between Fab-7D11 and L1 reveals the basis for the conformation-specific binding as recognition of a discontinuous epitope containing two loops that are held together by a disulfide bond. The structure of this important conformational epitope of L1 will contribute to the development of molecular poxvirus vaccines and also provides a novel target for anti-poxvirus drugs. In addition, the sequence and structure of Fab-7D11 will contribute to the development of L1-targeted immunotherapeutics.

An increase in melioidosis cases compared to other areas in Taiwan was observed in the Er-Ren River Basin, southwestern Taiwan, from November 2001 to August 2006. The objective of this study was to determine the association between the level of exposure to Burkholderia pseudomallei and the incidence rate of melioidosis and to survey the transmission modes of B. pseudomallei in the Er-Ren River Basin. The serosurveillance of melioidosis gave seropositivity rates of 36.6%, 21.6%, and 10.9%, respectively, for residents in regions A, B, and C within the Er-Ren Basin area. Culture and PCR-based detection of B. pseudomallei from soil demonstrated that the geographical distribution of this bacterium was confined to a particular site in region B. The distribution of seropositive titers was significantly associated with the incidence rate of melioidosis (120, 68, or 36 incidence cases per 100,000 population in region A, B, or C in 2005), whereas it did not correlate with the geographical distribution of B. pseudomallei within the soil. A survey of transmission modes showed that residents with seropositivity were linked to factors such as having confronted flooding and having walked barefoot on soil, which are potential risk factors associated with exposure to B. pseudomallei. Our findings indicated that the Er-Ren River Basin in Taiwan has the potential to become a high-prevalence area for melioidosis. This is the first report that documents a high prevalence of melioidosis in an area north of latitude 20 degrees N.