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1.
J Anim Ecol ; 92(5): 1089-1101, 2023 05.
Artículo en Inglés | MEDLINE | ID: mdl-36932966

RESUMEN

Range boundaries are long-term biogeographic features of species distributions and abundance. However, many species demonstrate dynamic range boundaries, reflecting strong seasonal and annual variability in migratory behaviour. As a form of facultative migration, irruptions involve the movement of many individuals outside of their resident range in response to climate variability, resource availability, and demographic processes. Many species have experienced range shifts and altered phenology in response to modern climate change, but spatiotemporal changes in irruption dynamics are less well known. We quantified changes in the geography and periodicity of boreal bird irruptions across eastern North America from 1960 to 2021. Using data from Audubon's Christmas Bird Count for nine finch species, including several exhibiting recent population declines, we evaluated latitudinal trends in southern range and irruption boundaries and characterized irruption periodicity using spectral wavelet analysis. Six boreal birds exhibited significant northward shifts in their southern range boundaries and three species displayed shifts in their southern irruption boundaries. Irruption periodicity across multiple species was consistent across the 1960s and 1970s, culminating in frequent and synchronized irruptions of multiple species (superflights) during earlier decades. Coherence between species dampened beginning in the early 1980s as superflight periodicity became increasingly unstructured, finally reforming in recent decades, after 2000. Boreal birds are considered important sentinels of the boreal forests, and northward shifts and altered periodicity of irruptions may indicate broad-scale changes in climate- and resource-associated drivers operating across the boreal forests.


Asunto(s)
Migración Animal , Aves , Animales , Aves/fisiología , Migración Animal/fisiología , Estaciones del Año , Cambio Climático , Geografía
3.
Conserv Biol ; 29(3): 784-94, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25627009

RESUMEN

The sustained decline in marine fisheries worldwide underscores the need to understand and monitor fisheries trends and fisher behavior. Recreational fisheries are unique in that they are not subject to the typical drivers that influence commercial and artisanal fisheries (e.g., markets or food security). Nevertheless, although exposed to a different set of drivers (i.e., interest or relaxation), recreational fisheries can contribute to fishery declines. Recreational fisheries are also difficult to assess due to an absence of past monitoring and traditional fisheries data. Therefore, we utilized a nontraditional data source (a chronology of spearfishing publications) to document historical trends in recreational spearfishing in Australia between 1952 and 2009. We extracted data on reported fish captures, advertising, and spearfisher commentary and used regression models and ordination analyses to assess historical change. The proportion of coastal fish captures reported declined approximately 80%, whereas the proportion of coral reef and pelagic fish reports increased 1750% and 560%, respectively. Catch composition shifted markedly from coastal temperate or subtropical fishes during the 1950s to 1970s to coral reef and pelagic species in the 1990s to 2000s. Advertising data and commentary by spearfishers indicated that pelagic fish species became desired targets. The mean weight of trophy coral reef fishes also declined significantly over the study period (from approximately 30-8 kg). Recreational fishing presents a highly dynamic social-ecological interface and a challenge for management. Our results emphasize the need for regulatory agencies to work closely with recreational fishing bodies to observe fisher behavior, detect shifts in target species or fishing intensity, and adapt regulatory measures.


Asunto(s)
Explotaciones Pesqueras/estadística & datos numéricos , Recreación , Animales , Australia , Conservación de los Recursos Naturales , Peces/fisiología , Dinámica Poblacional
4.
Cancer Res ; 74(18): 4955-66, 2014 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-25172841

RESUMEN

The ubiquitin-proteasome system (UPS) has emerged as a therapeutic focus and target for the treatment of cancer. The most clinically successful UPS-active agents (bortezomib and lenalidomide) are limited in application to hematologic malignancies, with only marginal efficacy in solid tumors. Inhibition of specific ubiquitin E3 ligases has also emerged as a valid therapeutic strategy, and many targets are currently being investigated. Another emerging and promising approach in regulation of the UPS involves targeting deubiquitinases (DUB). The DUBs comprise a relatively small group of proteins, most with cysteine protease activity that target several key proteins involved in regulation of tumorigenesis, apoptosis, senescence, and autophagy. Through their multiple contacts with ubiquitinated protein substrates involved in these pathways, DUBs provide an untapped means of modulating many important regulatory proteins that support oncogenic transformation and progression. Ubiquitin-specific proteases (USP) are one class of DUBs that have drawn special attention as cancer targets, as many are differentially expressed or activated in tumors or their microenvironment, making them ideal candidates for drug development. This review attempts to summarize the USPs implicated in different cancers, the current status of USP inhibitor-mediated pharmacologic intervention, and future prospects for USP inhibitors to treat diverse cancers.


Asunto(s)
Neoplasias/tratamiento farmacológico , Neoplasias/enzimología , Proteasas Ubiquitina-Específicas/metabolismo , Animales , Humanos , Terapia Molecular Dirigida , Inhibidores de Proteasas/uso terapéutico , Complejo de la Endopetidasa Proteasomal/metabolismo , Proteasas Ubiquitina-Específicas/antagonistas & inhibidores , Ubiquitinación/efectos de los fármacos
5.
World J Urol ; 31(6): 1327-32, 2013 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-23595605

RESUMEN

PURPOSE: To preliminarily evaluate the potential for an improvement in diagnostic performance by a combined interpretation of In-111 capromab pendetide single photon emission computed tomography (SPECT) including computed tomography (CT) image fusion with magnetic resonance diffusion-weighted imaging (MR-DWI) for identifying prostate cancer in pelvic lymph nodes thru correlation with histopathology. MATERIALS AND METHODS: This institutional approved, retrospective study identified patients with available histopathology of lymph nodes removed at the time of radical prostatectomy and who had undergone staging with In-111 capromab pendetide SPECT-CT and/or pelvic MRI (including DWI). The performance of In-111 capromab pendetide SPECT for identifying malignant lymph nodes was assessed. Subsequently, a combined reading of In-111 capromab pendetide SPECT and prostate MRI with DWI was performed and the performance assessed. RESULTS: 18 patients underwent In-111 capromab pendetide SPECT-CT. Of these, 12 patients had also undergone imaging with MR-DWI. In-111 capromab pendetide SPECT-CT had a sensitivity of 40.0% and specificity of 96.7% for identification of malignant lymph nodes. However, In-111 capromab pendetide SPECT-CT combined with MRI with DWI had a sensitivity of 88.9% and specificity of 98.5%. CONCLUSIONS: The addition of MR-DWI to the interpretation of In-111 capromab pendetide SPECT-CT may increase the sensitivity for detecting malignant lymph nodes in prostate cancer. Future prospective evaluation of combined In-111 capromab pendetide SPECT-CT and MR-DWI is indicated and may improve clinical evaluation of nodal disease in prostate cancer.


Asunto(s)
Anticuerpos Monoclonales , Imagen de Difusión por Resonancia Magnética , Radioisótopos de Indio , Neoplasias Pélvicas/diagnóstico , Neoplasias de la Próstata/patología , Tomografía Computarizada de Emisión de Fotón Único/métodos , Tomografía Computarizada por Rayos X/métodos , Anciano , Humanos , Ganglios Linfáticos/diagnóstico por imagen , Ganglios Linfáticos/patología , Metástasis Linfática/diagnóstico , Masculino , Persona de Mediana Edad , Neoplasias Pélvicas/secundario , Estudios Prospectivos , Próstata/diagnóstico por imagen , Próstata/patología , Neoplasias de la Próstata/diagnóstico por imagen , Estudios Retrospectivos , Sensibilidad y Especificidad
6.
Conserv Biol ; 27(2): 261-9, 2013 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-23140101

RESUMEN

We examined the development of coral reef science and the policies, institutions, and governance frameworks for management of coral reefs in China in order to highlight the wicked problem of preserving reefs while simultaneously promoting human development and nation building. China and other sovereign states in the region are experiencing unprecedented economic expansion, rapid population growth, mass migration, widespread coastal development, and loss of habitat. We analyzed a large, fragmented literature on the condition of coral reefs in China and the disputed territories of the South China Sea. We found that coral abundance has declined by at least 80% over the past 30 years on coastal fringing reefs along the Chinese mainland and adjoining Hainan Island. On offshore atolls and archipelagos claimed by 6 countries in the South China Sea, coral cover has declined from an average of >60% to around 20% within the past 10-15 years. Climate change has affected these reefs far less than coastal development, pollution, overfishing, and destructive fishing practices. Ironically, these widespread declines in the condition of reefs are unfolding as China's research and reef-management capacity are rapidly expanding. Before the loss of corals becomes irreversible, governance of China's coastal reefs could be improved by increasing public awareness of declining ecosystem services, by providing financial support for training of reef scientists and managers, by improving monitoring of coral reef dynamics and condition to better inform policy development, and by enforcing existing regulations that could protect coral reefs. In the South China Sea, changes in policy and legal frameworks, refinement of governance structures, and cooperation among neighboring countries are urgently needed to develop cooperative management of contested offshore reefs.


Asunto(s)
Conservación de los Recursos Naturales/legislación & jurisprudencia , Arrecifes de Coral , Política Ambiental/legislación & jurisprudencia , Animales , Antozoos/crecimiento & desarrollo , China , Cambio Climático , Ambiente , Explotaciones Pesqueras , Crecimiento Demográfico
7.
J Am Chem Soc ; 134(37): 15357-70, 2012 Sep 19.
Artículo en Inglés | MEDLINE | ID: mdl-22891849

RESUMEN

Incorporation of divalent metal ions into an active site is a fundamental catalytic tool used by diverse enzymes. Divalent cations are used by protein kinases to both stabilize ATP binding and accelerate chemistry. Kinetic analysis establishes that Cyclin-dependent kinase 2 (CDK2) requires simultaneous binding of two Mg(2+) ions for catalysis of phosphoryl transfer. This tool, however, comes with a price: the rate-acceleration effects are opposed by an unavoidable rate-limiting consequence of the use of two Mg(2+) ions by CDK2. The essential metal ions stabilize ADP product binding and limit the overall rate of the reaction. We demonstrate that product release is rate limiting for activated CDK2 and evaluate the effects of the two catalytically essential Mg(2+) ions on the stability of the ADP product within the active site. We present two new crystal structures of CDK2 bound to ADP showing how the phosphate groups can be coordinated by either one or two Mg(2+) ions, with the occupancy of one site in a weaker equilibrium. Molecular dynamics simulations indicate that ADP phosphate mobility is more restricted when ADP is coordinated by two Mg(2+) ions compared to one. The structural similarity between the rigid ADP·2Mg product and the cooperatively assembled transition state provides a mechanistic rational for the rate-limiting ADP release that is observed. We demonstrate that although the simultaneous binding of two Mg(2+) ions is essential for efficient phosphoryl transfer, the presence of both Mg(2+) ions in the active site also cooperatively increases ADP affinity and opposes its release. Evolution of protein kinases must have involved careful tuning of the affinity for the second Mg(2+) ion in order to balance the needs to stabilize the chemical transition state and allow timely product release. The link between Mg(2+) site affinity and activity presents a chemical handle that may be used by regulatory factors as well as explain some mutational effects.


Asunto(s)
Magnesio/química , Metales/química , Proteínas Quinasas/química , Calorimetría , Catálisis , Cristalografía por Rayos X , Modelos Moleculares , Simulación de Dinámica Molecular , Electricidad Estática
8.
Structure ; 19(5): 675-90, 2011 May 11.
Artículo en Inglés | MEDLINE | ID: mdl-21565702

RESUMEN

We have determined high-resolution crystal structures of a CDK2/Cyclin A transition state complex bound to ADP, substrate peptide, and MgF(3)(-). Compared to previous structures of active CDK2, the catalytic subunit of the kinase adopts a more closed conformation around the active site and now allows observation of a second Mg(2+) ion in the active site. Coupled with a strong [Mg(2+)] effect on in vitro kinase activity, the structures suggest that the transient binding of the second Mg(2+) ion is necessary to achieve maximum rate enhancement of the chemical reaction, and Mg(2+) concentration could represent an important regulator of CDK2 activity in vivo. Molecular dynamics simulations illustrate how the simultaneous binding of substrate peptide, ATP, and two Mg(2+) ions is able to induce a more rigid and closed organization of the active site that functions to orient the phosphates, stabilize the buildup of negative charge, and shield the subsequently activated γ-phosphate from solvent.


Asunto(s)
Adenosina Difosfato/metabolismo , Ciclina A/química , Quinasa 2 Dependiente de la Ciclina/química , Fluoruros/metabolismo , Compuestos de Magnesio/metabolismo , Proteínas Recombinantes/química , Adenosina Trifosfato/metabolismo , Biocatálisis , Dominio Catalítico , Clonación Molecular , Cristalización , Cristalografía por Rayos X , Ciclina A/genética , Ciclina A/metabolismo , Quinasa 2 Dependiente de la Ciclina/genética , Quinasa 2 Dependiente de la Ciclina/metabolismo , Activación Enzimática , Escherichia coli , Humanos , Cinética , Modelos Moleculares , Simulación de Dinámica Molecular , Fosfatos/metabolismo , Unión Proteica , Conformación Proteica , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Especificidad por Sustrato
9.
Mol Endocrinol ; 24(8): 1650-64, 2010 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-20610536

RESUMEN

Thyroid hormone receptor (TR), as a member of the nuclear hormone receptor family, can recognize and bind different classes of DNA response element targets as either a monomer, a homooligomer, or a heterooligomer. We report here the first crystal structure of a homodimer TR DNA-binding domain (DBD) in complex with an inverted repeat class of thyroid response element (TRE). The structure shows a nearly symmetric structure of the TR DBD assembled on the F2 TRE where the base recognition contacts in the homodimer DNA complex are conserved relative to the previously published structure of a TR-9-cis-retinoic acid receptor heterodimer DNA complex. The new structure also reveals that the T-box region of the DBD can function as a structural hinge that enables a large degree of flexibility in the position of the C-terminal extension helix that connects the DBD to the ligand-binding domain. Although the isolated TR DBDs exist as monomers in solution, we have measured highly cooperative binding of the two TR DBD subunits onto the inverted repeat DNA sequence. This suggests that elements of the DBD can influence the specific TR oligomerization at target genes, and it is not just interactions between the ligand-binding domains that are responsible for TR oligomerization at target genes. Mutational analysis shows that intersubunit contacts at the DBD C terminus account for some, but not all, of the cooperative homodimer TR binding to the inverted repeat class TRE.


Asunto(s)
Receptores de Hormona Tiroidea/química , Receptores de Hormona Tiroidea/metabolismo , Elementos de Respuesta/genética , Animales , Polarización de Fluorescencia , Unión Proteica/genética , Unión Proteica/fisiología , Multimerización de Proteína/genética , Multimerización de Proteína/fisiología , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína/genética , Estructura Terciaria de Proteína/fisiología , Ratas , Receptores de Hormona Tiroidea/genética , Inversión de Secuencia/genética , Inversión de Secuencia/fisiología
10.
FEBS J ; 275(13): 3397-407, 2008 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-18494797

RESUMEN

We have shown that hormone-induced downregulation of luteinizing hormone receptor (LHR) in the ovary is post-transcriptionally regulated by an mRNA binding protein. This protein, later identified as mevalonate kinase (MVK), binds to the coding region of LHR mRNA, suppresses its translation, and the resulting ribonucleoprotein complex is targeted for degradation. Mutagenesis and crystallographic studies of rat MVK have established Ser146, Glu193, Asp204 and Lys13 as being crucial for its catalytic function. The present study examined the structural aspects of MVK required for LHR mRNA recognition and translational suppression. Single MVK mutants (S146A, E193Q, D204N and K13A) were overexpressed in 293T cells. Cytosolic fractions were examined for LHR mRNA binding activities by RNA electrophoretic mobility shift analysis. All the single MVK mutants showed decreased LHR mRNA binding activity compared with the wild-type MVK. Double mutants (S146A & E193Q, E193Q & D204N and E193Q & K13A) of MVK also showed a significant decrease in binding to LHR mRNA, suggesting that the residues required for catalytic function are also involved in LHR mRNA recognition. Mutation of the residues outside the catalytic site (D316A and S314A) did not cause any change in LHR mRNA binding activity of MVK when compared with wild-type MVK. To examine the biological effects of these mutants on LHR mRNA expression, a full-length capped rat LHR mRNA was synthesized and translated using a rabbit reticulocyte lysate system in the presence or absence of the MVK mutant proteins. The results showed that mutations of the active site residues of MVK abrogated the inhibitory effect on LHR mRNA translation. Therefore, these data indicate that an intact active site of MVK is required for its binding to rat LHR mRNA and for its translational suppressor function.


Asunto(s)
Regulación Enzimológica de la Expresión Génica , Hormona Luteinizante/metabolismo , Fosfotransferasas (Aceptor de Grupo Alcohol)/fisiología , Receptores de HL/química , Animales , Dominio Catalítico , Cristalografía por Rayos X , Humanos , Mutación , Fosfotransferasas (Aceptor de Grupo Alcohol)/metabolismo , Unión Proteica , ARN Mensajero/metabolismo , Conejos , Ratas , Reticulocitos/metabolismo
11.
Langmuir ; 22(25): 10354-64, 2006 Dec 05.
Artículo en Inglés | MEDLINE | ID: mdl-17129003

RESUMEN

Electrochemical surface-enhanced hyper-Raman scattering (SEHRS) and surface-enhanced Raman scattering (SERS) of centrosymmetric molecules on Ag film over nanosphere (AgFON) electrodes are presented. The SEHR spectra of trans-1,2-bis(4-pyridyl)ethylene (BPE) at different potentials (vs Ag/AgCl) are presented for the first time, and a reversible potential tuning of the SEHR spectra of BPE is demonstrated. The SEHRS and SERS techniques were used to determine to what extent either site symmetry reduction or field gradient effects dictate the origin of the observed vibrational spectra. It is found that the SEHR and SER spectra for the molecules studied were distinctly different at all frequency regions at a fixed voltage, suggesting that centrosymmetry is largely retained upon adsorption to the AgFON surface and that field gradient effects are negligible. This work also shows that the SEHR spectra clearly depend on potential, whereas the SER spectra are essentially independent of potenial. It is determined that the combination of changes in deltaGads and the presence of coadsorbed counterions are responsible for altering the local symmetry of the adsorbate and only SEHRS has the sensitivity to detect these changes in the surface environment. Thus, SEHRS is a uniquely useful spectroscopic tool that is much more sensitive to the local adsorption environment than is SERS.


Asunto(s)
Membranas Artificiales , Nanotubos/química , Plata/química , Adsorción , Electrodos , Dispersión de Radiación , Sensibilidad y Especificidad , Espectrometría Raman/métodos , Propiedades de Superficie
12.
Faraday Discuss ; 132: 9-26, 2006.
Artículo en Inglés | MEDLINE | ID: mdl-16833104

RESUMEN

Surface-enhanced Raman spectroscopy (SERS) is currently experiencing a renaissance in its development driven by the remarkable discovery of single molecule SERS (SMSERS) and the explosion of interest in nanophotonics and plasmonics. Because excitation of the localized surface plasmon resonance (LSPR) of a nanostructured surface or nanoparticle lies at the heart of SERS, it is important to control all of the factors influencing the LSPR in order to maximize signal strength and ensure reproducibility. These factors include material, size, shape, interparticle spacing, and dielectric environment. All of these factors must be carefully controlled to ensure that the incident laser light maximally excites the LSPR in a reproducible manner. This article describes the use of nanosphere lithography for the fabrication of highly reproducible and robust SERS substrates for both fundamental studies and applications. Atomic layer deposition (ALD) is introduced as a novel fabrication method for dielectric spacers to study the SERS distance dependence and control the nanoscale dielectric environment. Wavelength scanned SER excitation spectroscopy (WS SERES) measurements show that enhancement factors approximately 10(8) are obtainable from NSL-fabricated surfaces and provide new insight into the electromagneticfield enhancement mechanism. Tip-enhanced Raman spectroscopy (TERS) is an extremely promising new development to improve the generality and information content of SERS. A 2D correlation analysis is applied to SMSERS data. Finally, the first in vivo SERS glucose sensing study is presented.


Asunto(s)
Espectrometría Raman , Técnicas Biosensibles/instrumentación , Glucosa/análisis , Nanopartículas , Nanotubos , Espectrometría Raman/instrumentación , Espectrometría Raman/métodos , Resonancia por Plasmón de Superficie , Propiedades de Superficie
13.
PLoS Biol ; 4(5): e144, 2006 May.
Artículo en Inglés | MEDLINE | ID: mdl-16640460

RESUMEN

The improper activation of the Abl tyrosine kinase results in chronic myeloid leukemia (CML). The recognition of an inactive conformation of Abl, in which a catalytically important Asp-Phe-Gly (DFG) motif is flipped by approximately 180 degrees with respect to the active conformation, underlies the specificity of the cancer drug imatinib, which is used to treat CML. The DFG motif is not flipped in crystal structures of inactive forms of the closely related Src kinases, and imatinib does not inhibit c-Src. We present a structure of the kinase domain of Abl, determined in complex with an ATP-peptide conjugate, in which the protein adopts an inactive conformation that resembles closely that of the Src kinases. An interesting aspect of the Src-like inactive structure, suggested by molecular dynamics simulations and additional crystal structures, is the presence of features that might facilitate the flip of the DFG motif by providing room for the phenylalanine to move and by coordinating the aspartate side chain as it leaves the active site. One class of mutations in BCR-Abl that confers resistance to imatinib appears more likely to destabilize the inactive Src-like conformation than the active or imatinib-bound conformations. Our results suggest that interconversion between distinctly different inactive conformations is a characteristic feature of the Abl kinase domain.


Asunto(s)
Proteínas Proto-Oncogénicas c-abl/química , Familia-src Quinasas/química , Secuencias de Aminoácidos , Antineoplásicos/farmacología , Benzamidas , Sitios de Unión , Resistencia a Antineoplásicos , Activación Enzimática , Humanos , Mesilato de Imatinib , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Modelos Moleculares , Piperazinas/farmacología , Unión Proteica , Estructura Terciaria de Proteína , Proteínas Proto-Oncogénicas c-abl/metabolismo , Pirimidinas/farmacología
14.
Cancer Res ; 66(2): 1007-14, 2006 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-16424036

RESUMEN

We present a high-resolution (2.0 A) crystal structure of the catalytic domain of a mutant form of the Abl tyrosine kinase (H396P; Abl-1a numbering) that is resistant to the Abl inhibitor imatinib. The structure is determined in complex with the small-molecule inhibitor VX-680 (Vertex Pharmaceuticals, Cambridge, MA), which blocks the activity of various imatinib-resistant mutant forms of Abl, including one (T315I) that is resistant to both imatinib and BMS-354825 (dasatinib), a dual Src/Abl inhibitor that seems to be clinically effective against all other imatinib-resistant forms of BCR-Abl. VX-680 is shown to have significant inhibitory activity against BCR-Abl bearing the T315I mutation in patient-derived samples. The Abl kinase domain bound to VX-680 is not phosphorylated on the activation loop in the crystal structure but is nevertheless in an active conformation, previously unobserved for Abl and inconsistent with the binding of imatinib. The adoption of an active conformation is most likely the result of synergy between the His(396)Pro mutation, which destabilizes the inactive conformation required for imatinib binding, and the binding of VX-680, which favors the active conformation through hydrogen bonding and steric effects. VX-680 is bound to Abl in a mode that accommodates the substitution of isoleucine for threonine at residue 315 (the "gatekeeper" position). The avoidance of the innermost cavity of the Abl kinase domain by VX-680 and the specific recognition of the active conformation explain the effectiveness of this compound against mutant forms of BCR-Abl, including those with mutations at the gatekeeper position.


Asunto(s)
Proteínas de Fusión bcr-abl/genética , Genes abl , Piperazinas/farmacología , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas c-abl/química , Pirimidinas/farmacología , Aurora Quinasas , Benzamidas , Dominio Catalítico , Cristalografía , Dasatinib , Resistencia a Antineoplásicos , Escherichia coli/genética , Humanos , Enlace de Hidrógeno , Mesilato de Imatinib , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Fosforilación , Conformación Proteica , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-abl/metabolismo , Tiazoles/farmacología
15.
J Am Chem Soc ; 127(12): 4484-9, 2005 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-15783231

RESUMEN

A rapid detection protocol suitable for use by first-responders to detect anthrax spores using a low-cost, battery-powered, portable Raman spectrometer has been developed. Bacillus subtilis spores, harmless simulants for Bacillus anthracis, were studied using surface-enhanced Raman spectroscopy (SERS) on silver film over nanosphere (AgFON) substrates. Calcium dipicolinate (CaDPA), a biomarker for bacillus spores, was efficiently extracted by sonication in nitric acid and rapidly detected by SERS. AgFON surfaces optimized for 750 nm laser excitation have been fabricated and characterized by UV-vis diffuse reflectance spectroscopy and SERS. The SERS signal from extracted CaDPA was measured over the spore concentration range of 10(-14)-10(-12) M to determine the saturation binding capacity of the AgFON surface and to calculate the adsorption constant (Kspore=1.7 x 10(13) M(-1)). At present, an 11 min procedure is capable of achieving a limit of detection (LOD) of approximately 2.6 x 10(3) spores, below the anthrax infectious dose of 10(4) spores. The data presented herein also demonstrate that the shelf life of prefabricated AgFON substrates can be as long as 40 days prior to use. Finally, these sensing capabilities have been successfully transitioned from a laboratory spectrometer to a field-portable instrument. Using this technology, 10(4) bacillus spores were detected with a 5 s data acquisition period on a 1 month old AgFON substrate. The speed and sensitivity of this SERS sensor indicate that this technology can be used as a viable option for the field analysis of potentially harmful environmental samples.


Asunto(s)
Bacillus anthracis/química , Ácidos Picolínicos/análisis , Espectrometría Raman/métodos , Biomarcadores/análisis , Calcio/análisis , Nanoestructuras/química , Plata/química , Esporas Bacterianas/química
16.
J Phys Chem B ; 109(22): 11279-85, 2005 Jun 09.
Artículo en Inglés | MEDLINE | ID: mdl-16852377

RESUMEN

A detailed wavelength-scanned surface-enhanced Raman excitation spectroscopy (WS SERES) study of benzenethiol adsorbed on Ag nanoparticle arrays, fabricated by nanosphere lithography (NSL), is presented. These NSL-derived Ag nanoparticle array surfaces are both structurally well-characterized and extremely uniform in size. The WS SERES spectra are correlated, both spatially and spectrally, with the corresponding localized surface plasmon resonance (LSPR) spectra of the nanoparticle arrays. The surface-enhanced Raman scattering (SERS) spectra were measured in two excitation wavelength ranges: (1) 425-505 nm, and (2) 610-800 nm, as well as with the 532-nm line from a solid-state diode-pumped laser. The WS SERES spectra have line shapes similar to those of the LSPR spectra. The maximum SERS enhancement factor is shown to occur for excitation wavelengths that are blue-shifted with respect to the LSPR lambda(max) of adsorbate-covered nanoparticle arrays. Three vibrational modes of benzenethiol (1575, 1081, and 1009 cm(-1)) are studied simultaneously on one substrate, and it is demonstrated that the smaller Raman shifted peak shows a maximum enhancement closer to the LSPR lambda(max) than that of a larger Raman shifted peak. This is in agreement with the predictions of the electromagnetic (EM) enhancement mechanism of SERS. Enhancement factors of up to approximately 10(8) are achieved, which is also in good agreement with our previous SERES studies.


Asunto(s)
Espectrometría Raman/métodos , Ciclohexanos/química , Campos Electromagnéticos , Microesferas , Nanopartículas , Fenoles/química , Fotones , Espectrometría Raman/instrumentación , Compuestos de Sulfhidrilo/química , Propiedades de Superficie
17.
Biophys J ; 87(6): 3799-813, 2004 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-15326025

RESUMEN

We describe herein a computationally intensive project aimed at carrying out molecular dynamics (MD) simulations including water and counterions on B-DNA oligomers containing all 136 unique tetranucleotide base sequences. This initiative was undertaken by an international collaborative effort involving nine research groups, the "Ascona B-DNA Consortium" (ABC). Calculations were carried out on the 136 cases imbedded in 39 DNA oligomers with repeating tetranucleotide sequences, capped on both ends by GC pairs and each having a total length of 15 nucleotide pairs. All MD simulations were carried out using a well-defined protocol, the AMBER suite of programs, and the parm94 force field. Phase I of the ABC project involves a total of approximately 0.6 mus of simulation for systems containing approximately 24,000 atoms. The resulting trajectories involve 600,000 coordinate sets and represent approximately 400 gigabytes of data. In this article, the research design, details of the simulation protocol, informatics issues, and the organization of the results into a web-accessible database are described. Preliminary results from 15-ns MD trajectories are presented for the d(CpG) step in its 10 unique sequence contexts, and issues of stability and convergence, the extent of quasiergodic problems, and the possibility of long-lived conformational substates are discussed.


Asunto(s)
ADN/química , Modelos Químicos , Modelos Moleculares , Oligodesoxirribonucleótidos/química , Secuencia de Bases , Simulación por Computador , Cinética , Datos de Secuencia Molecular , Movimiento (Física) , Conformación de Ácido Nucleico , Relación Estructura-Actividad
18.
Cell ; 112(6): 859-71, 2003 Mar 21.
Artículo en Inglés | MEDLINE | ID: mdl-12654251

RESUMEN

c-Abl is normally regulated by an autoinhibitory mechanism, the disruption of which leads to chronic myelogenous leukemia. The details of this mechanism have been elusive because c-Abl lacks a phosphotyrosine residue that triggers the assembly of the autoinhibited form of the closely related Src kinases by internally engaging the SH2 domain. Crystal structures of c-Abl show that the N-terminal myristoyl modification of c-Abl 1b binds to the kinase domain and induces conformational changes that allow the SH2 and SH3 domains to dock onto it. Autoinhibited c-Abl forms an assembly that is strikingly similar to that of inactive Src kinases but with specific differences that explain the differential ability of the drug STI-571/Gleevec/imatinib (STI-571) to inhibit the catalytic activity of Abl, but not that of c-Src.


Asunto(s)
Proteínas Proto-Oncogénicas c-abl/química , Benzamidas , Catálisis , Cristalografía por Rayos X , Inhibidores Enzimáticos/farmacología , Mesilato de Imatinib , Modelos Moleculares , Fosforilación , Piperazinas/farmacología , Unión Proteica , Conformación Proteica , Proteínas Proto-Oncogénicas c-abl/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-abl/aislamiento & purificación , Proteínas Proto-Oncogénicas c-abl/metabolismo , Piridinas , Pirimidinas/química , Pirimidinas/farmacología , Proteínas Recombinantes de Fusión/metabolismo , Relación Estructura-Actividad , Dominios Homologos src
19.
Biopolymers ; 68(1): 3-15, 2003 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-12579576

RESUMEN

To assess the accuracy of the molecular dynamics (MD) models of nucleic acids, a detailed comparison between MD-calculated and NMR-observed indices of the dynamical structure of DNA in solution has been carried out. The specific focus of our comparison is the oligonucleotide duplex, d(CGCGAATTCGCG)(2), for which considerable structural data have been obtained from crystallography and NMR spectroscopy. An MD model for the structure of d(CGCGAATTCGCG)(2) in solution, based on the AMBER force field, has been extended with a 14 ns trajectory. New NMR data for this sequence have been obtained in order to allow a detailed and critical comparison between the calculated and observed parameters. Observable two-dimensional (2D) nuclear Overhauser effect spectroscopy (NOESY) volumes and scalar coupling constants were back-calculated from the MD trajectory and compared with the corresponding NMR data. The comparison of these results indicate that the MD model is in generally good agreement with the NMR data, and shows closer accord with experiment than back-calculations based on the crystal structure of d(CGCGAATTCGCG)(2) or the canonical A or B forms of the sequence. The NMR parameters are not particularly sensitive to the known deficiency in the AMBER MD model, which is a tendency toward undertwisting of the double helix when the parm.94 force field is used. The MD results are also compared with a new determination of the solution structure of d(CGCGAATTCGCG)(2) using NMR dipolar coupling data.


Asunto(s)
ADN/química , Resonancia Magnética Nuclear Biomolecular , Conformación de Ácido Nucleico , Secuencia de Bases , Cristalización , ADN/genética , Modelos Moleculares , Soluciones/química
20.
Cancer Res ; 62(15): 4244-55, 2002 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-12154026

RESUMEN

The early stage of chronic myelogenous leukemia (CML) is caused by the tyrosine kinase Bcr-Abl. Imatinib mesylate (also known as STI-571 and Gleevec), a tyrosine kinase inhibitor, has shown encouraging results in CML clinical trials and has become a paradigm for targeted cancer therapeutics. Recent reports of resistance to imatinib argue for further development of therapies for CML. During studies of signal transduction, we observed that the pyrido[2,3-d]pyrimidine src tyrosine kinase inhibitor PD173955 inhibited Bcr-Abl-dependent cell growth. Subsequently, a related compound, PD180970, was reported as a potent inhibitor of Bcr-Abl. We have compared the potency of these two compounds and four other analogues with imatinib on Bcr-Abl-dependent cell growth, cytokine-dependent cell growth, and tyrosine kinase inhibition. PD173955 inhibited Bcr-Abl-dependent cell growth with an IC(50) of 2-35 nM in different cell lines. Fluorescence-activated cell-sorting analyses of cells treated with PD173955 showed cell cycle arrest in G(1). PD173955 has an IC(50) of 1-2 nM in kinase inhibition assays of Bcr-Abl, and in cellular growth assays it inhibits Bcr-Abl-dependent substrate tyrosine phosphorylation. Of the six pyrido[2,3-d]pyrimidine analogues studied, PD166326 was the most potent inhibitor of Bcr-Abl-dependent cell growth. PD173955 inhibited kit ligand-dependent c-kit autophosphorylation (IC(50) = approximately 25 nM) and kit ligand-dependent proliferation of M07e cells (IC(50) = 40 nM) but had a lesser effect on interleukin 3-dependent (IC(50) = 250 nM) or granulocyte macrophage colony-stimulating factor (IC(50) = 1 microM)-dependent cell growth. These compounds are potent inhibitors of both the Bcr-Abl and c-kit receptor tyrosine kinases and deserve further study as potential treatments for both CML and for diseases in which c-kit has a role.


Asunto(s)
Inhibidores Enzimáticos/farmacología , Proteínas Tirosina Quinasas/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-kit/efectos de los fármacos , Piridonas/farmacología , Pirimidinas/farmacología , Proteínas Tirosina Quinasas Receptoras/antagonistas & inhibidores , División Celular/efectos de los fármacos , División Celular/fisiología , Inhibidores Enzimáticos/química , Proteínas de Fusión bcr-abl , Fase G1/efectos de los fármacos , Células Madre Hematopoyéticas/efectos de los fármacos , Células Madre Hematopoyéticas/enzimología , Células Madre Hematopoyéticas/patología , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Leucemia Mielógena Crónica BCR-ABL Positiva/enzimología , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Modelos Moleculares , Células Madre Neoplásicas/efectos de los fármacos , Células Madre Neoplásicas/enzimología , Células Madre Neoplásicas/patología , Fosforilación/efectos de los fármacos , Proteínas Tirosina Quinasas/fisiología , Proteínas Proto-Oncogénicas c-kit/metabolismo , Piridonas/química , Pirimidinas/química , Proteínas Tirosina Quinasas Receptoras/metabolismo , Relación Estructura-Actividad
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