20 results on '"Piechowicz M"'
Search Results
2. 352 Gpr143 and dopamine signalling control lysosomal activity in retinal pigment epithelial cells
- Author
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Piechowicz, M and McKay, BS
- Published
- 2018
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3. A report on emergent uranyl binding phenomena by an amidoxime phosphonic acid co-polymer.
- Author
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Abney, C. W., Das, S., Mayes, R. T., Kuo, L.-J., Wood, J., Gill, G., Piechowicz, M., Lin, Z., Lin, W., and Dai, S.
- Abstract
The development of technology to harvest the uranium dissolved in seawater would enable access to vast quantities of this critical metal for nuclear power generation. Amidoxime polymers are the most promising platforms for achieving this separation, yet the design of advanced adsorbents is hindered by uncertainty regarding the uranium binding mode. In this work we use XAFS to investigate the uranium coordination environment in an amidoxime–phosphonic acid copolymer adsorbent. In contrast to the binding mode predicted computationally and from small molecule studies, a cooperative chelating model is favoured, attributable to emergent behavior resulting from inclusion of amidoxime in the polymer. Samples exposed to seawater also display a feature consistent with a μ
2 -oxo-bridged transition metal, suggesting the formation of an in situ specific binding site. These findings challenge long held assumptions and provide new opportunities for the design of advanced adsorbent materials. [ABSTRACT FROM AUTHOR]- Published
- 2016
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4. XAFS investigation of polyamidoxime-bound uranyl contests the paradigm from small molecule studies.
- Author
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Abney, C. W., Mayes, R. T., Piechowicz, M., Lin, Z., Bryantsev, V. S., Veith, G. M., Dai, S., and Lin, W.
- Published
- 2016
- Full Text
- View/download PDF
5. Effect of peer-education on the willingness to vaccinate against COVID-19 among high school students.
- Author
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Pasek, O., Michalska, J., Piechowicz, M., Stoliński, M., and Ganczak, M.
- Subjects
AFFINITY groups ,VACCINATION ,COVID-19 ,ATTITUDE (Psychology) ,COVID-19 vaccines ,CONFERENCES & conventions ,HIGH school students ,EDUCATIONAL outcomes - Abstract
Introduction: The willingness to vaccinate against COVID-19 among adolescents remains low. Peer-education about SARS-CoV-2 may positively influence attitudes to vaccinate among this group. Objective: To assess the influence of peer-based educational intervention on the willingness to vaccinate against COVID-19 among Polish high school students. Methods: In the Polish Lubuskie province, a peer education campaign was introduced among final year high school students from 24 randomly selected schools. Before and after the 45-minute lecture, conducted online by 13 medical students in 51 classes with science and other programs, the willingness to vaccinate was checked using an anonymous questionnaire. The p-value was calculated with the use of chi-square and Wilcoxon signed rank tests. Results: Out of 883 students, 638 responded (72%); 268 males (42%), mean age: 18.4±0.6 years, 69% lived in cities with <100,000 inhabitants, 32% were in the science program. The willingness to vaccinate improved from 31.8% to 35.2% after education (p<0.001); in females from 29.7% to 33.1%; in males from 34% to 37.7% (p<0.002; p<0.01 respectively). Before intervention 43.5% students living in the large cities and 26.5% in small cities wanted to be vaccinated, this increased to 48.0% and 29.5% respectively (p<0.003; p<0.005) after intervention. There were significant between-locations and between-programs differences in the willingness to vaccinate before and after intervention (p<0.001; p<0.001 and p<0.001; p<0.001 respectively). The improvement in the science program was from 44.3% to 47.8%; in the other programs from 26.1% to 29.5% (p<0.01; p<0.002 respectively). Conclusions: Adolescents’ willingness to vaccinate against COVID-19 was alarmingly low, especially among females, students from small cities and attending the non-science program. Concise integrated teaching designed to address mistrust and knowledge deficiencies can be effective in improving the uptake. Key messages: Research provide data about students’ attitudes towards COVID-19 vaccination and indicate the need for universal educational campaigns. Online education is an effective method which may substantially increase COVID-19 vaccination rate in Poland. [ABSTRACT FROM AUTHOR]
- Published
- 2021
6. Low androgen/progesterone or high oestrogen/androgen receptors ratio in serous ovarian cancer predicts longer survival.
- Author
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Gogola-Mruk J, Pietrus M, Piechowicz M, Milian-Ciesielska K, Głód P, Wolnicka-Glubisz A, Szpor J, and Ptak A
- Subjects
- Humans, Female, Cell Line, Tumor, Estrogen Receptor alpha metabolism, Estrogen Receptor alpha genetics, Middle Aged, Prognosis, Cystadenocarcinoma, Serous metabolism, Cystadenocarcinoma, Serous pathology, Cystadenocarcinoma, Serous mortality, Gene Expression Regulation, Neoplastic, Ovarian Neoplasms metabolism, Ovarian Neoplasms mortality, Receptors, Androgen metabolism, Receptors, Androgen genetics, Receptors, Progesterone metabolism, Receptors, Progesterone genetics
- Abstract
The treatment of ovarian cancer (OC) remains one of the greatest challenges in gynaecological oncology. The presence of classic steroid receptors in OC makes hormone therapy an attractive option; however, the response of OC to hormone therapy is modest. Here, we compared the expression patterns of progesterone (PGR), androgen (AR) and oestrogen alpha (ERα) receptors between serous OC cell lines and non-cancer ovarian cells. These data were analysed in relation to steroid receptor expression profiles from patient tumour samples and survival outcomes using a bioinformatics approach. The results showed that ERα, PGR and AR were co-expressed in OC cell lines, and patient samples from high-grade and low-grade OC co-expressed at least two steroid receptors. High AR expression was negatively correlated, whereas ERα and PGR expression was positively correlated with patient survival. AR showed the opposite expression pattern to that of ERα and PGR in type 1 (SKOV-3) and 2 (OVCAR-3) OC cell lines compared with non-cancer (HOSEpiC) ovarian cells, with AR downregulated in type 1 and upregulated in type 2 OC. A low AR/PGR ratio and a high ESR1/AR ratio were associated with favourable survival outcomes in OC compared with other receptor ratios. Although the results must be interpreted with caution because of the small number of primary tumour samples analysed, they nevertheless suggest that the evaluation of ERα, AR and PGR by immunohistochemistry should be performed in patient biological material to plan future clinical trials., 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 © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2024
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7. Preventing and treating delirium in clinical settings for older adults.
- Author
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Faeder M, Hale E, Hedayati D, Israel A, Moschenross D, Peterson M, Peterson R, Piechowicz M, Punzi J, and Gopalan P
- Abstract
Delirium is a serious consequence of many acute or worsening chronic medical conditions, a side effect of medications, and a precipitant of worsening functional and cognitive status in older adults. It is a syndrome characterized by fluctuations in cognition and impaired attention that develops over a short period of time in response to an underlying medical condition, a substance (prescribed, over the counter, or recreational), or substance withdrawal and can be multi-factorial. We present a narrative review of the literature on nonpharmacologic and pharmacologic approaches to prevention and treatment of delirium with a focus on older adults as a vulnerable population. Older adult patients are most at risk due to decreasing physiologic reserves, with delirium rates of up to 80% in critical care settings. Presentation of delirium can be hyperactive, hypoactive, or mixed, making identification and study challenging as patients with hypoactive delirium are less likely to come to attention in an inpatient or long-term care setting. Studies of delirium focus on prevention and treatment with nonpharmacological or medication interventions, with the preponderance of evidence favoring multi-component nonpharmacological approaches to prevention as the most effective. Though use of antipsychotic medication in delirium is common, existing evidence does not support routine use, showing no clear benefit in clinically significant outcome measures and with evidence of harm in some studies. We therefore suggest that antipsychotics be used to treat agitation, psychosis, and distress associated with delirium at the lowest effective doses and shortest possible duration and not be considered a treatment of delirium itself. Future studies may clarify the use of other agents, such as melatonin and melatonin receptor agonists, alpha-2 receptor agonists, and anti-epileptics., Competing Interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s), 2023.)
- Published
- 2023
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8. Correction: A modified aeroponic system for growing small-seeded legumes and other plants to study root systems.
- Author
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Cai J, Veerappan V, Arildsen K, Sullivan C, Piechowicz M, Frugoli J, and Dickstein R
- Published
- 2023
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9. A modified aeroponic system for growing small-seeded legumes and other plants to study root systems.
- Author
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Cai J, Veerappan V, Arildsen K, Sullivan C, Piechowicz M, Frugoli J, and Dickstein R
- Abstract
Background: Various growth systems are available for studying plant root growth and plant-microbe interactions including hydroponics and aeroponics. Although some of these systems work well with Arabidopsis thaliana and smaller cereal model plants, they may not scale up as well for use with hundreds of plants at a time from a larger plant species. The aim of this study is to present step-by-step instructions for fabricating an aeroponic system, also called a "caisson," that has been in use in several legume research labs studying the development of symbiotic nitrogen fixing nodules, but for which detailed directions are not currently available. The aeroponic system is reusable and is adaptable for many other types of investigations besides root nodulation., Results: An aeroponic system that is affordable and reusable was adapted from a design invented by French engineer René Odorico. It consists of two main components: a modified trash can with a lid of holes and a commercially available industrial humidifier that is waterproofed with silicon sealant. The humidifier generates a mist in which plant roots grow, suspended from holes in trash can lid. Results from use of the aeroponic system have been available in the scientific community for decades; it has a record as a workhorse in the lab., Conclusions: Aeroponic systems present a convenient way for researchers to grow plants for studying root systems and plant-microbe interactions in root systems. They are particularly attractive for phenotyping roots and following the progress of nodule development in legumes. Advantages include the ability to precisely control the growth medium in which the plants grow and easy observations of roots during growth. In this system, mechanical shear potentially killing microbes found in some other types of aeroponic devices is not an issue. Disadvantages of aeroponic systems include the likelihood of altered root physiology compared to root growth on soil and other solid substrates and the need to have separate aeroponic systems for comparing plant responses to different microbial strains., (© 2023. The Author(s).)
- Published
- 2023
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10. Impact of Water Extraction on Malonamide Aggregation: A Molecular Dynamics and Graph Theoretic Approach.
- Author
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Servis MJ, Piechowicz M, and Soderholm L
- Subjects
- Malonates, Solutions, Molecular Dynamics Simulation, Water
- Abstract
Solution structure in liquid-liquid extraction affects the efficacy of separation; however, even for simplified organic phases, structural characterization and attribution of aggregation to intermolecular interactions are fundamental challenges. We investigate water uptake into organic phases for two malonamides commonly applied to actinide and lanthanide separations. Extracted water induces reorganization of the amphiphilic extractant molecules, although we find this rearrangement is not strongly manifested in small-angle X-ray scattering making it challenging to probe without methods such as atomistic simulation. Using a graph theoretic approach to define hydrogen bonded water/malonamide aggregates from molecular dynamics simulations, we find evidence of a characteristic aggregate size by water number that results from geometric accommodation of the surrounding malonamide molecules. This implies a degree of size selectivity inherent to these water-in-oil aggregates. Conversely, we find no evidence of a characteristic size of the aggregates with respect to their malonamide number. By defining a separate graphical representation of self-association of the amphiphilic malonamides, we quantify how water affects the local and nonlocal topology of the malonamide network, providing a basis for characterization of the structure and impact of polar solutes in increasingly complex organic phases.
- Published
- 2021
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11. Molecular-scale origins of solution nanostructure and excess thermodynamic properties in a water/amphiphile mixture.
- Author
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Servis MJ, Piechowicz M, Skanthakumar S, and Soderholm L
- Abstract
The molecular and nanoscale origins of nonideality in excess thermodynamic properties are essential to understanding cosolvent mixtures, yet they remain challenging to determine. Here, we consider a binary mixture of water and an amphiphile, N,N,N',N'-tetramethylmalonamide (TMMA), which is characterized by strong hydrogen bonding between the two components and no hydrogen bonding between amphiphiles. Using molecular dynamics simulation, validated with excess volume measurements and X-ray scattering, we identify three distinct solution regimes across the composition range of the binary mixture and find that the transition between two of these regimes, marked by the water percolation threshold, is closely correlated with minima in the excess volume and excess enthalpy. Structural analysis of the simulations reveals an interplay between local interactions and solution nanostructure, determined by the relative strength of the water-water and water-amphiphile hydrogen bonding interactions. By comparison with other amphiphiles, such as linear alcohols, the relative strength of like and unlike interactions between water and amphiphile affects the relationship between thermodynamics and structural regimes. This provides insight into how molecular forces of mutual solvation interact across length scales and how they manifest in excess thermodynamic properties.
- Published
- 2021
- Full Text
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12. Amphiphile Organization in Organic Solutions: An Alternative Explanation for Small-Angle X-ray Scattering Features in Malonamide/Alkane Mixtures.
- Author
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Servis MJ, Piechowicz M, Shkrob IA, Soderholm L, and Clark AE
- Abstract
The role of different intermolecular interactions in the aggregation of amphiphiles in an organic solvent is studied for systems of relevance to liquid-liquid extraction (LLE), a chemical process used to selectively recover metals from complex mixtures. Of specific interest is the role, or lack thereof, of hydrogen bonding, which is often assumed to be a main driver of the organic phase structural organization that has been linked to separation efficacy. Toward that end, a series of malonamide extractants in n -dodecane have been studied in the absence of any extracted aqueous solutes, including water. The series of extractants includes N , N '-dimethyl- N , N '-dibutyltetradecylmalonamide (DMDBTDMA), two of its homologs, and N , N '-dimethyl- N , N '-dioctylhexylethoxymalonamide (DMDOHEMA). This simplified model LLE system enables systematic investigation of the role of dipole-dipole and alkyl tail steric interactions in amphiphile aggregation. Small-angle X-ray scattering (SAXS) profiles computed from molecular dynamics trajectories are in good agreement with experimental SAXS data. Molecular dynamics simulations show that malonamide aggregation results from dipole-driven self-association and lacks characteristic aggregate sizes. Mid- q correlation peaks in the SAXS profiles emerge at high concentration for each malonamide. In those densely packed solutions, the correlation peaks are observed to result from alkyl tail-induced spacing between electron-rich polar head groups, with peak positions determined by the different alkyl tail lengths present in the malonamide molecule. This explanation of the SAXS correlation peaks contrasts with the prevailing literature, which attributes mesoscale features observed in small-angle scattering to the formation of microemulsions. Instead, this work finds that these features are present in the absence of water or any reverse micellar organization of the malonamides. As such, molecular-scale malonamide self-association and packing, rather than microemulsion-based colloidal-scale descriptions, is a more appropriate framework for these LLE systems.
- Published
- 2020
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13. Leveraging Actinide Hydrolysis Chemistry for Targeted Th and U Separations using Amidoxime-Functionalized Poly(HIPE)s.
- Author
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Piechowicz M, Chiarizia R, Skanthakumar S, Rowan SJ, and Soderholm L
- Abstract
Polymerized high internal phase emulsions (poly(HIPE)s) are porous polymer monoliths whose synthesis can easily be tailored to allow incorporation of functional units. In this work, nitrile containing poly(HIPE)s have been prepared with either acrylonitrile (AN) or 4-cyanostyrene (4CS) comonomers. Post-synthetic modification of these nitrile-containing poly(HIPE)s yields their corresponding amidoximated analogues, which were studied for actinide uptake. These amidoxime-functionalized, porous polymers were shown to adsorb 95 % Th
4+ species from aqueous solution within 30 minutes. In contrast to other amidoxime containing polymers the uptake of UO2 2+ in these poly(HIPE)s is lower under similar conditions. A critical analysis of actinide separations and high-energy X-ray scattering data provides insight into the polymers' selectivity, enabled by the uptake of multinuclear Th clusters., (© 2020 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)- Published
- 2020
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14. Metastatic and non-metastatic sentinel inguinofemoral lymph nodes in vulvar cancer show an increased lymphangiogenesis.
- Author
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Piechowicz M, Mikos M, Banas T, Okon K, Pietrus M, Balajewicz-Nowak M, Szczudlik L, Kojs Z, Czerw A, Juszczyk G, and Pityński K
- Subjects
- Aged, Animals, Antibodies, Monoclonal, Murine-Derived, Carcinoma, Squamous Cell pathology, Female, Groin, Humans, Immunohistochemistry, Mice, Neoplasm Staging, Prospective Studies, Rosaniline Dyes, Technetium Tc 99m Sulfur Colloid, Vulvar Neoplasms pathology, Carcinoma, Squamous Cell diagnosis, Lymphangiogenesis, Lymphatic Metastasis diagnosis, Sentinel Lymph Node pathology, Vulvar Neoplasms diagnosis
- Abstract
Introduction and Objective: Lymph node involvement is a strong predictor of disease recurrence and patient survival in vulvar cancer. The aim of the study was to evaluate the feasibility of sentinel lymph node (SLN) screening, the incidence of skip metastases, and lymph node lymphangiogenesis., Material and Methods: Fifty-five patients participated in this prospective, single centre study. A double SLN screening method was employed using radiocolloid (technetium-99 sulfur colloid) and 1.0% Isosulfan Blue. Immunohistochemistry, using a mouse monoclonal antibody against D2-40, was used to evaluate lymphatic vessel density (LVD). All calculations were performed using STATISTICA software v. 10 (StatSoft, USA, 2011); p < 0.05 was considered significant., Results: Using both methods of SLN detection, 100% accuracy was achieved, and skip metastases were diagnosed in only one woman (1.82%). Peri-tumour median LVD was significantly increased compared with matched intra-tumour samples (p < 0.001), while median LVD was significantly lower in negative, compared with positive SLN, regardless of whether matched non-SLN were negative (p < 0.001) or positive (p = 0.005). Metastatic SLN exhibited significantly higher median LVD compared with matched negative non-SLN (p = 0.015), while no significant difference in median LVD was detected between positive SLN and matched positive non-SLN. However, negative SLN had a significantly higher median LVD compared with matched negative non-SLN (p = 0.012)., Conclusions: SLN detection is a safe and feasible procedure in vulvar cancer. In patients without nodular involvement, SLN, compared with non-SLN, exhibited significantly higher median LVD, which may be an indication of its preparation to host metastases, and thus requires further investigation.
- Published
- 2020
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15. Disruption of MET Receptor Tyrosine Kinase, an Autism Risk Factor, Impairs Developmental Synaptic Plasticity in the Hippocampus.
- Author
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Ma X, Chen K, Lu Z, Piechowicz M, Liu Q, Wu J, and Qiu S
- Subjects
- Age Factors, Animals, Animals, Newborn, Enzyme Inhibitors pharmacology, Female, Hippocampus drug effects, Homeodomain Proteins genetics, Homeodomain Proteins metabolism, In Vitro Techniques, Indoles pharmacology, Lysine analogs & derivatives, Lysine metabolism, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Nerve Net drug effects, Nerve Net physiology, Neuronal Plasticity drug effects, Neurons drug effects, Proto-Oncogene Proteins c-met genetics, RNA Interference physiology, Signal Transduction drug effects, Signal Transduction genetics, Sulfones pharmacology, Transcription Factors genetics, Transcription Factors metabolism, Hippocampus cytology, Neuronal Plasticity physiology, Neurons physiology, Proto-Oncogene Proteins c-met metabolism
- Abstract
As more genes conferring risks to neurodevelopmental disorders are identified, translating these genetic risk factors into biological mechanisms that impact the trajectory of the developing brain is a critical next step. Here, we report that disrupted signaling mediated MET receptor tyrosine kinase (RTK), an established risk factor for autism spectrum disorders, in the developing hippocampus glutamatergic circuit leads to profound deficits in neural development, synaptic transmission, and plasticity. In cultured hippocampus slices prepared from neonatal mice, pharmacological inhibition of MET kinase activity suppresses dendritic arborization and disrupts normal dendritic spine development. In addition, single-neuron knockdown (RNAi) or overexpression of Met in the developing hippocampal CA1 neurons leads to alterations, opposite in nature, in basal synaptic transmission and long-term plasticity. In forebrain-specific Met conditional knockout mice (Met
fx/fx ;emx1cre ), an enhanced long-term potentiation (LTP) and long-term depression (LTD) were observed at early developmental stages (P12-14) at the Schaffer collateral to CA1 synapses compared with wild-type littermates. In contrast, LTP and LTD were markedly reduced at young adult stage (P56-70) during which wild-type mice show robust LTP and LTD. The altered trajectory of synaptic plasticity revealed by this study indicate that temporally regulated MET signaling as an intrinsic, cell autonomous, and pleiotropic mechanism not only critical for neuronal growth and functional maturation, but also for the timing of synaptic plasticity during forebrain glutamatergic circuits development., (© 2018 Wiley Periodicals, Inc.)- Published
- 2019
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16. A Versatile Colorimetric Probe based on Thiosemicarbazide-Amine Proton Transfer.
- Author
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Calvino C, Piechowicz M, Rowan SJ, Schrettl S, and Weder C
- Abstract
There is significant interest in the rapid and efficient detection of amines, which are widely used in different industries and also serve as markers in many biological processes. This work reports that the coupling of a thiosemicarbazide binding motif and a naphthalimide-based chromophore affords highly sensitive sensor molecules, which can indicate the presence of amines with a pronounced and readily visible color change. It was demonstrated that the binding mechanism involves a proton transfer from the thiosemicarbazide to the analyte. This process renders the mechanism highly sensitive and broadly exploitable. The potential usefulness of the sensor is demonstrated by fabricating an indicator paper, which allows for the detection of volatile amines at concentrations as low as ca. 10 ppm., (© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2018
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17. Insight into selectivity: uptake studies of radionuclides 90 Sr 2+ , 137 Cs + , and 233 UO 2 2+ with bis-amidoxime polymers.
- Author
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Piechowicz M, Chiarizia R, and Soderholm L
- Abstract
Uptake characteristics of a bis-amidoximated polymer are presented for 90Sr2+, 137Cs+, and 233UO22+ to assess rational ligand design and polymer engineering efforts applied to selective uranium extraction. Functionalized with the bis-amidoxime diaryl ether ligand at a loading of 1.98 mmol per gram of polymer, the polymer was found to sorb uranium from a pH 6 solution with a separation factor (α) over cesium of 1.2 × 103. Strontium uptake was negligible throughout the pH range studied whereas cesium uptake averaged <30%. Moreover, at near neutral pH, the sorbent was able to purify water to below the United States' set maximum contaminant level for uranium, decreasing the U concentration from 330 ppb to below 3 ppb in 30 minutes. Density functional theory (DFT) calculations, used to probe the nature of the metal-bis-amidoxime interaction, help inform the empirically realized uptake selectivity.
- Published
- 2018
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18. Successful Coupling of a Bis-Amidoxime Uranophile with a Hydrophilic Backbone for Selective Uranium Sequestration.
- Author
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Piechowicz M, Abney CW, Thacker NC, Gilhula JC, Wang Y, Veroneau SS, Hu A, and Lin W
- Abstract
The amidoxime group (-RNH
2 NOH) has long been used to extract uranium from seawater on account of its high affinity toward uranium. The development of tunable sorbent materials for uranium sequestration remains a research priority as well as a significant challenge. Herein, we report the design, synthesis, and uranium sorption properties of bis-amidoxime-functionalized polymeric materials (BAP 1-3). Bifunctional amidoxime monomers were copolymerized with an acrylamide cross-linker to obtain bis-amidoxime incorporation as high as 2 mmol g-1 after five synthetic steps. The resulting sorbents were able to uptake nearly 600 mg of uranium per gram of polymer after 37 days of contact with a seawater simulant containing 8 ppm uranium. Moreover, the polymeric materials exhibited low vanadium uptake with a maximum capacity of 128 mg of vanadium per gram of polymer. This computationally predicted and experimentally realized selectivity of uranium over vanadium, nearly 5 to 1 w/w, is one of the highest reported to date and represents an advancement in the rational design of sorbent materials with high uptake capacity and selectivity.- Published
- 2017
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19. A Simplified Method for Ultra-Low Density, Long-Term Primary Hippocampal Neuron Culture.
- Author
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Lu Z, Piechowicz M, and Qiu S
- Subjects
- Animals, Cells, Cultured, Mice, Neurons physiology, Hippocampus cytology, Neurons cytology, Primary Cell Culture methods
- Abstract
Culturing primary hippocampal neurons in vitro facilitates mechanistic interrogation of many aspects of neuronal development. Dissociated embryonic hippocampal neurons can often grow successfully on glass coverslips at high density under serum-free conditions, but low density cultures typically require a supply of trophic factors by co-culturing them with a glia feeder layer, preparation of which can be time-consuming and laborious. In addition, the presence of glia may confound interpretation of results and preclude studies on neuron-specific mechanisms. Here, a simplified method is presented for ultra-low density (~2,000 neurons/cm2), long-term (>3 months) primary hippocampal neuron culture that is under serum free conditions and without glia cell support. Low density neurons are grown on poly-D-lysine coated coverslips, and flipped on high density neurons grown in a 24-well plate. Instead of using paraffin dots to create a space between the two neuronal layers, the experimenters can simply etch the plastic bottom of the well, on which the high density neurons reside, to create a microspace conducive to low density neuron growth. The co-culture can be easily maintained for >3 months without significant loss of low density neurons, thus facilitating the morphological and physiological study of these neurons. To illustrate this successful culture condition, data are provided to show profuse synapse formation in low density cells after prolonged culture. This co-culture system also facilitates the survival of sparse individual neurons grown in islands of poly-D-lysine substrates and thus the formation of autaptic connections.
- Published
- 2016
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20. Plasticity of binocularity and visual acuity are differentially limited by nogo receptor.
- Author
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Stephany CÉ, Chan LL, Parivash SN, Dorton HM, Piechowicz M, Qiu S, and McGee AW
- Subjects
- Animals, GPI-Linked Proteins metabolism, Immunohistochemistry, Mice, Mice, Inbred C57BL, Mice, Knockout, Microscopy, Confocal, Neurogenesis physiology, Nogo Receptor 1, Parvalbumins metabolism, Patch-Clamp Techniques, Interneurons metabolism, Myelin Proteins metabolism, Neuronal Plasticity physiology, Receptors, Cell Surface metabolism, Vision, Binocular physiology, Visual Acuity physiology
- Abstract
The closure of developmental critical periods consolidates neural circuitry but also limits recovery from early abnormal sensory experience. Degrading vision by one eye throughout a critical period both perturbs ocular dominance (OD) in primary visual cortex and impairs visual acuity permanently. Yet understanding how binocularity and visual acuity interrelate has proven elusive. Here we demonstrate the plasticity of binocularity and acuity are separable and differentially regulated by the neuronal nogo receptor 1 (NgR1). Mice lacking NgR1 display developmental OD plasticity as adults and their visual acuity spontaneously improves after prolonged monocular deprivation. Restricting deletion of NgR1 to either cortical interneurons or a subclass of parvalbumin (PV)-positive interneurons alters intralaminar synaptic connectivity in visual cortex and prevents closure of the critical period for OD plasticity. However, loss of NgR1 in PV neurons does not rescue deficits in acuity induced by chronic visual deprivation. Thus, NgR1 functions with PV interneurons to limit plasticity of binocularity, but its expression is required more extensively within brain circuitry to limit improvement of visual acuity following chronic deprivation., (Copyright © 2014 the authors 0270-6474/14/3411631-10$15.00/0.)
- Published
- 2014
- Full Text
- View/download PDF
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