bluuebunny/crossref_metadata_2025_split huggingface dataset:
Reactivation of Aggregated Proteins by the ClpB/DnaK Bi‐Chaperone System
Authors: Zolkiewski Michal , Chesnokova Liudmila S. , Witt Stephan N. 
Published: February 2016

AbstractProtein aggregation is a common problem in protein biochemistry and is linked to many cellular pathologies and human diseases. The molecular chaperone ClpB can resolubilize and reactivate aggregated proteins. This unit describes the procedure for following reactivation of an aggregated enzyme glucose‐6‐phosphate dehydrogenase mediated by ClpB from Escherichia coli in cooperation with another molecular chaperone, DnaK. The procedures for purification of these chaperones are also described. © 2016 by John Wiley & Sons, Inc.
Expression, Solubilization, and Purification of Bacterial Membrane Proteins
Authors: Jeffery Constance J. 
Published: February 2016

AbstractBacterial integral membrane proteins play many important roles, including sensing changes in the environment, transporting molecules into and out of the cell, and in the case of commensal or pathogenic bacteria, interacting with the host organism. Working with membrane proteins in the lab can be more challenging than working with soluble proteins because of difficulties in their recombinant expression and purification. This protocol describes a standard method to express, solubilize, and purify bacterial integral membrane proteins. The recombinant protein of interest with a 6His affinity tag is expressed in E. coli. After harvesting the cultures and isolating cellular membranes, mild detergents are used to solubilize the membrane proteins. Protein‐detergent complexes are then purified using IMAC column chromatography. Support protocols are included to help select a detergent for protein solubilization and for use of gel filtration chromatography for further purification. © 2016 by John Wiley & Sons, Inc.
Isolation of Human Basophils
Authors: Schroeder John T. , Bieneman Anja P. 
Published: February 2016

AbstractIsolating human basophils from blood has long been hampered by the fact that these granulocytes represent just 1% or less of the circulating leukocyte population. We describe herein laboratory protocols that have been refined over the past ∼25 years that now enable investigators to prepare basophils for use in a variety of assays to assess the in vitro biology of these immune cells, both in IgE ‐dependent and ‐independent responses. © 2016 by John Wiley & Sons, Inc.
Measuring Apoptosis by Microscopy and Flow Cytometry
Authors: Hollville Emilie , Martin Seamus J. 
Published: February 2016

AbstractApoptosis is a mode of programmed cell death that plays an important role during development and in the maintenance of tissue homeostasis. Numerous physiological as well as pathological stimuli trigger apoptosis such as engagement of Fas, TRAIL, or TNF receptors, growth factor deprivation, hypoxia, or exposure to cytotoxic drugs. Apoptosis is coordinated from within by members of the caspase family of cysteine proteases that, upon activation, trigger a series of morphological changes including cell shrinkage, extensive plasma membrane blebbing, chromatin condensation, DNA hydrolysis, and nuclear fragmentation. These dramatic structural and biochemical alterations result not only in the controlled dismantling of the cell, but also in the efficient recognition and removal of apoptotic cells by phagocytes. Necrosis, which is typically nonprogrammed or imposed upon the cell by overwhelming membrane or organelle damage, is characterized by rapid plasma membrane rupture followed by organelle and cell swelling. Necrosis is often provoked by infectious agents or a severe departure from physiological conditions. This unit describes protocols for the measurement of apoptosis and for distinguishing apoptosis from necrosis. © 2016 by John Wiley & Sons, Inc.
Culture, Expansion, and Differentiation of Murine Megakaryocytes from Fetal Liver, Bone Marrow, and Spleen
Authors: Schulze Harald 
Published: February 2016

AbstractMegakaryocytes (MKs) are the source of circulating platelets and are readily recognized by their large size and distinctive morphology. Their poor representation in hematopoietic tissues often requires considerable ex vivo expansion to generate cells for biochemical and cell biological studies. These experimental protocols describe the assessment of megakaryocytic potential within hematopoietic precursor cells in the bone marrow by colony‐forming assays and expansion and enrichment of MKs from cultured fetal liver or spleen or bone marrow cells. Although these MKs are not synchronized in their maturation, they can be enriched over an albumin step gradient, and one‐third to one‐half of recovered cells will typically elaborate proplatelets, the immediate precursors of blood platelets. Both protocols require recombinant thrombopoietin (TPO) as a growth factor. Support protocols describe methods for preparing fetal liver cells, identifying mature rodent MKs by staining for acetylcholinesterase activity, and staining (May‐Grünwald‐Giemsa) mixed populations on cytocentrifuged blood cell preparations. © 2016 by John Wiley & Sons, Inc.
Setting constraints on the nature and origin of the two major hydrous sulfates on Mars: Monohydrated and polyhydrated sulfates
Authors: Wang Alian , Jolliff Bradley L. , Liu Yang , Connor Kathryn 
Published: February 2016

AbstractMonohydrated Mg sulfate (MgSO4·H2O) and polyhydrated sulfate are the most common and abundant hydrous sulfates observed thus far on Mars. They are widely distributed and coexist in many locations. On the basis of results from two new sets of experiments, in combination with past experimental studies and the subsurface salt mineralogy observed at a saline playa (Dalangtan, DLT) in a terrestrial analogue hyperarid region on the Tibet Plateau, we can now set new constraints on the nature and origin of these two major Martian sulfates. Starkeyite (MgSO4·4H2O) is the best candidate for polyhydrated sulfate. MgSO4·H2O in the form of “LH‐1w,” generated from dehydration of Mg sulfates with high degrees of hydration, is the most likely mineral form for the majority of Martian monohydrated Mg sulfate. Two critical properties of Mg sulfates are responsible for the coexistence of these two phases that have very different degrees of hydration: (1) the metastability of a substructural unit in starkeyite at relatively low temperatures, and (2) catalytic effects attributed to coprecipitated species (sulfates, chlorides, oxides, and hydroxides) from chemically complex brines that help overcome the metastability of starkeyite. The combination of these two properties controls the coexistence of the LH‐1w layer and starkeyite layers at many locations on Mars, which sometimes occur in an interbedded stratigraphy. The structural H2O held by these two broadly distributed sulfates represents a large H2O reservoir at the surface and in the shallow subsurface on current Mars.
Groundwater transit time distribution and mean from streambed sampling in an agricultural coastal plain watershed, North Carolina, USA
Authors: Gilmore Troy E. , Genereux David P. , Solomon D. Kip , Solder John E. 
Published: February 2016

AbstractWe measured groundwater apparent age (τ) and seepage rate (v) in a sandy streambed using point‐scale sampling and seepage blankets (a novel seepage meter). We found very similar MTT estimates from streambed point sampling in a 58 m reach (29 years) and a 2.5 km reach (31 years). The TTD for groundwater discharging to the stream was best fit by a gamma distribution model and was very similar for streambed point sampling in both reaches. Between adjacent point‐scale and seepage blanket samples, water from the seepage blankets was generally younger, largely because blanket samples contained a fraction of “young” stream water. Correcting blanket data for the stream water fraction brought τ estimates for most blanket samples closer to those for adjacent point samples. The MTT estimates from corrected blanket data were in good agreement with those from sampling streambed points adjacent to the blankets. Collectively, agreement among age‐dating tracers, general accord between tracer data and piston‐flow model curves, and large groundwater age gradients in the streambed, suggested that the piston flow apparent ages were reasonable estimates of the groundwater transit times for most samples. Overall, our results from two field campaigns suggest that groundwater collected in the streambed can provide reasonable estimates of apparent age of groundwater discharge, and that MTT can be determined from different age‐dating tracers and by sampling with different groundwater collection devices. Coupled streambed point measurements of groundwater age and groundwater seepage rate represent a novel, reproducible, and effective approach to estimating aquifer TTD and MTT.
Improving the theoretical underpinnings of process‐based hydrologic models
Authors: Clark Martyn P. , Schaefli Bettina , Schymanski Stanislaus J. , Samaniego Luis , Luce Charles H. , Jackson Bethanna M. , Freer Jim E. , Arnold Jeffrey R. , Moore R. Dan , Istanbulluoglu Erkan , Ceola Serena 
Published: February 2016

AbstractIn this Commentary, we argue that it is possible to improve the physical realism of hydrologic models by making better use of existing hydrologic theory. We address the following questions: (1) what are some key elements of current hydrologic theory; (2) how can those elements best be incorporated where they may be missing in current models; and (3) how can we evaluate competing hydrologic theories across scales and locations? We propose that hydrologic science would benefit from a model‐based community synthesis effort to reframe, integrate, and evaluate different explanations of hydrologic behavior, and provide a controlled avenue to find where understanding falls short.
Influence of soil and climate on root zone storage capacity
Authors: de Boer‐Euser Tanja , McMillan Hilary K. , Hrachowitz Markus , Winsemius Hessel C. , Savenije Hubert H. G. 
Published: February 2016

AbstractRoot zone storage capacity (Sr) is an important variable for hydrology and climate studies, as it strongly influences the hydrological functioning of a catchment and, via evaporation, the local climate. Despite its importance, it remains difficult to obtain a well‐founded catchment representative estimate. This study tests the hypothesis that vegetation adapts its Sr to create a buffer large enough to sustain the plant during drought conditions of a certain critical strength (with a certain probability of exceedance). Following this method, Sr can be estimated from precipitation and evaporative demand data. The results of this “climate‐based method” are compared with traditional estimates from soil data for 32 catchments in New Zealand. The results show that the differences between catchments in climate‐derived catchment representative Sr values are larger than for soil‐derived Sr values. Using a model experiment, we show that the climate‐derived Sr can better reproduce hydrological regime signatures for humid catchments; for more arid catchments, the soil and climate methods perform similarly. This makes the climate‐based Sr a valuable addition for increasing hydrological understanding and reducing hydrological model uncertainty.
Potential for long‐lead prediction of the western North Pacific monsoon circulation beyond seasonal time scales
Authors: Choi Jung , Son Seok‐Woo , Seo Kyong‐Hwan , Lee June‐Yi , Kang Hyun‐Suk 
Published: February 2016

AbstractAlthough the western North Pacific (WNP) monsoon circulation significantly impacts the socioeconomic communities around Asia, its prediction is only limited to a few months. By examining the Coupled Model Intercomparison Project phase 5 decadal hindcast experiments, we explore a possibility of the extended prediction skill for the WNP monsoon circulation beyond seasonal time scales. It is found that the multimodel ensemble (MME) predictions, initialized in January, successfully predict the WNP circulation in spring and early summer. Somewhat surprisingly, a reliable prediction of the WNP circulation appears even in the following spring with a maximum lead time of 14 months. This unexpected prediction skill is likely caused by the improved El Niño–Southern Oscillation (ENSO) prediction and the exaggerated dynamical link between the ENSO and premonsoon circulation in the MME prediction. Although further studies are needed, this result may open up new opportunities for the multiseasonal prediction of the WNP monsoon circulation.
Ambroxol chaperone therapy for neuronopathic Gaucher disease: A pilot study
Authors: Narita Aya , Shirai Kentarou , Itamura Shinji , Matsuda Atsue , Ishihara Akiko , Matsushita Kumi , Fukuda Chisako , Kubota Norika , Takayama Rumiko , Shigematsu Hideo , Hayashi Anri , Kumada Tomohiro , Yuge Kotaro , Watanabe Yoriko , Kosugi Saori , Nishida Hiroshi , Kimura Yukiko , Endo Yusuke , Higaki Katsumi , Nanba Eiji , Nishimura Yoko , Tamasaki Akiko , Togawa Masami , Saito Yoshiaki , Maegaki Yoshihiro , Ohno Kousaku , Suzuki Yoshiyuki 
Published: February 2016

AbstractObjectiveGaucher disease (GD) is a lysosomal storage disease characterized by a deficiency of glucocerebrosidase. Although enzyme‐replacement and substrate‐reduction therapies are available, their efficacies in treating the neurological manifestations of GD are negligible. Pharmacological chaperone therapy is hypothesized to offer a new strategy for treating the neurological manifestations of this disease. Specifically, ambroxol, a commonly used expectorant, has been proposed as a candidate pharmacological chaperone. The purpose of this study was to evaluate the safety, tolerability, and neurological efficacy of ambroxol in patients with neuronopathic GD.MethodsThis open‐label pilot study included five patients who received high‐dose oral ambroxol in combination with enzyme replacement therapy. Safety was assessed by adverse event query, physical examination, electrocardiography, laboratory studies, and drug concentration. Biochemical efficacy was assessed through evidence of glucocerebrosidase activity in the lymphocytes and glucosylsphingosine levels in the cerebrospinal fluid. Neurological efficacy was evaluated using the Unified Myoclonus Rating Scale, Gross Motor Function Measure, Functional Independence Measure, seizure frequency, pupillary light reflex, horizontal saccadic latency, and electrophysiologic studies.ResultsHigh‐dose oral ambroxol had good safety and tolerability, significantly increased lymphocyte glucocerebrosidase activity, permeated the blood–brain barrier, and decreased glucosylsphingosine levels in the cerebrospinal fluid. Myoclonus, seizures, and pupillary light reflex dysfunction markedly improved in all patients. Relief from myoclonus led to impressive recovery of gross motor function in two patients, allowing them to walk again.InterpretationPharmacological chaperone therapy with high‐dose oral ambroxol shows promise in treating neuronopathic GD, necessitating further clinical trials.
Mesoporous Silica as Nanoreactors to Prepare Gd‐Encapsulated Carbon Dots of Controllable Sizes and Magnetic Properties
Authors: Chen Hongmin , Wang Geoffrey D. , Sun Xilin , Todd Trever , Zhang Fan , Xie Jin , Shen Baozhong 
Published: February 2016

Gd‐encapsulated carbonaceous dots (Gd@C‐dots) hold great potential in clinical applications as a novel type of T1 contrast agent for magnetic resonance imaging (MRI). However, current synthetic methods require multiple purification steps due to poor size control, making them unsuitable for high throughput. Herein, a novel, mesoporous silica nanoparticle (MSN)‐templated method for the size‐controlled synthesis of Gd@C‐dots is reported. Briefly, MSNs nanoreactors of different pore sizes are loaded with Gd precursors. Upon calcination, carbon layers are grown around the Gd cations. The spatial restraint of the silica cavity facilitates size control of the produced Gd@C‐dots. Specifically, using 3, 7, and 11 nm MSNs as templates allows the synthesis of 3.0, 7.4, and 9.6 nm Gd@C‐dots, respectively. A significant size impact on the magnetic and optical properties of the nanoparticles is shown, with the smallest Gd@C‐dots showing the highest r1 relaxivity (10 mM−1 s−1) and fluorescence quantum yield (30.2%). The 3.0‐nm Gd@C‐dots were then conjugated with a tumor‐targeting ligand, c(RGDyK), and injected into U87MG xenograft tumor models. Good tumor targeting was observed in T1‐weighted MRI images; whereby the unbound nanoparticles were efficiently excreted through renal clearance, avoiding long‐term toxicity to the host.
Contorted Arene‐fused Metallophthalocyanines
Authors: Suzuki Hiroyuki , Kobayashi Nagao , Kimura Mutsumi 
Published: February 2016

AbstractThree phthalonitriles 9–11 fused with planar and nonplanar polycyclic aromatic hydrocarbons were synthesized through a combination of palladium‐catalyzed coupling, electrophilic cyclization, and Scholl oxidation reactions. The equilibrium geometries and electronic structures of the frontier orbitals of 9–11 were simulated by computation calculations. The optimized structure of 11 is found to have a larger torsional angle than that of 10 because of the non‐planarity arising from some cove and fjord regions. Zinc complexes 1–3 fused with polycyclic aromatic hydrocarbons were synthesized by the cyclotetramerization of 9–11. The attachment of nonplanar polycyclic aromatic hydrocarbons with phthalocyanine rings prevented the formation of aggregates in the solid state.
Metal‐Free Direct Nucleophilic Perfluoroalkylthiolation with Perfluoroalkanesulfenamides
Authors: Glenadel Quentin , Bordy Mathieu , Alazet Sébastien , Tlili Anis , Billard Thierry 
Published: February 2016

AbstractWe report herein a practical method to generate CF3S− and RFS− anions in situ from shelf‐stable reagents, which were initially designed for electrophilic reactions. With an “iodide activation” step, these in situ released anions can directly participate in metal‐free nucleophilic substitution reactions with the loss of various leaving groups. Our method is compatible with various functional groups and provides good yields. With this strategy, the first direct nucleophilic perfluoroalkylthiolation reactions have been described.
The association between airline flight and sinonasal symptoms
Authors: Shargorodsky Josef , Zheng Laura , Stillman Frances , Soong Andrea , Navas‐Acien Ana , Reh Douglas 
Published: February 2016

BackgroundAirplane cabin supply air has been shown to contain multiple possible respiratory irritants. In addition, changes in barometric pressure in flight may contribute to specific respiratory conditions. Therefore, there may be an association between commercial airline flight and sinus disease.MethodsParticipants of the Secondhand‐Smoke, Air Quality and Respiratory Health Among Flight Attendants Study were administered an online questionnaire pertaining to their flight experience and respiratory health. Working years, working days per month, and number of trips per month were quantified, as well as smoking exposure and self‐reported physician diagnoses of sinusitis, asthma, and rhinitis. The sinonasal outcomes were quantified using a Respiratory Questionnaire Survey (RQS) score. Multivariable analyses were performed to evaluate the associations between flight time and sinus disease.ResultsA total of 579 participants met the inclusion criteria for this study, with cohort prevalence of sinusitis, asthma, and rhinitis of 25.3%, 14.4%, and 20.5%, respectively. Tertiles 2 and 3 of working days per month were associated with higher RQS scores compared to tertile 1 (p for trend <0.01). Individual symptoms significantly associated with increasing number of working days per month included “need to blow nose,” “sneezing,” and “thick nasal discharge,” and the number of international trips per month was significantly associated with “coughing” and “facial pain and pressure,” among other symptoms.ConclusionThis is the largest study to analyze the relations between airline flight time and sinonasal disease. The results suggest a possible association between sinusitis diagnosis, symptom scores, and specific sinonasal symptoms, and airline flight time.
Patient preferences for route of allergy immunotherapy: a comparison of four delivery methods
Authors: Chester Jennifer G. , Bremberg Maria G. , Reisacher William R. 
Published: February 2016

BackgroundAllergen‐specific immunotherapy (IT) is a disease‐modifying treatment for allergic rhinitis, and adherence to the treatment schedule is important for a successful outcome. Several methods for delivering IT are now available, but little information is known concerning patient preference for these options.MethodsOver a 12‐month period, 228 adults scheduled to undergo outpatient allergy testing were offered a survey to rank 4 different IT delivery methods: subcutaneous IT (SCIT), sublingual IT using liquid extracts (SLIT), sublingual allergy immunotherapy tablets (AIT), and oral mucosal immunotherapy (OMIT) using a toothpaste delivery vehicle.ResultsOf the participants who completed the survey, ranking scores (mean ± SD) for SCIT, SLIT, AIT, and OMIT, with 1 being the highest rank, were 3.36 ± 1.02, 2.03 ± 0.92, 2.30 ± 1.02, and 2.32 ± 1.05, respectively. The number of participants who ranked SCIT, SLIT, AIT, or OMIT as their number 1 choice was 24 (10.5%), 79 (34.6%), 61 (26.8%), and 64 (28.1%), respectively. When comparing first choice rankings, there were no statistically significant differences between SLIT, AIT, or OMIT, yet all 3 groups ranked higher than SCIT (p < 0.0001).ConclusionSCIT earned the lowest mean rank and had the least number of participants rank it as the most preferred method. Among the other 3 choices, by mean scores, SLIT was preferred most compared to either AIT or OMIT; but when considering first choices only, there were no significant differences in preference. Most of the study participants identified the convenience of home IT administration as the most important factor in their ranking.
Draf III mucosal graft technique: long‐term results
Authors: Illing Elisa A. , Cho Do Yeon , Riley Kristen O. , Woodworth Bradford A. 
Published: February 2016

BackgroundThe Draf III procedure is an effective endoscopic approach to the treatment of frontal sinus disorders, but has a significant restenosis rate. The objective of the current study is to assess long‐term success of the Draf III mucosal grafting technique.MethodsPatients undergoing Draf III with mucosal grafting technique were prospectively enrolled. Demographics, indication for surgery, anterior‐posterior (AP) diameter of frontal ostium, and complications were recorded. Failure was defined a priori as closure ≥50% of the intraoperative AP diameter at last clinical follow‐up.ResultsNinety‐six patients requiring Draf III with mucosal grafting were enrolled, 67 of which (average age 54 years; range, 15–84 years) had at least 1 year of follow‐up (average 34 months; range, 12–85 months) and were included in the analysis. Reasons for the procedure included chronic rhinosinusitis with frontal ostium stenosis (n = 37), tumor (n = 26), and cerebrospinal fluid leak (n = 4). Average preoperative Lund‐Mackay score was 13.9 ± 7.8. Average intraoperative AP diameter was 11.0 ± 1.9 mm. The procedure was highly effective with 97% (65/67) of patients maintaining a patent frontal sinus ostium (>50% intraoperative AP diameter) for the duration of follow‐up (average postoperative diameter 9.9 ± 2.2 mm). However, the 2 Draf III failures remained patent, did not need further intervention, and were considered clinically successful. Three patients required reoperation for reasons unrelated to closure of the Draf III.ConclusionCommon causes of failure following Draf III procedures include osteoneogenesis and stenosis. This study provides long‐term data demonstrating excellent outcomes using the mucosal grafting technique.