2,731 research outputs found
Experimental Methods in Cryogenic Spectroscopy: Stark Effect Measurements in Substituted Myoglobin
Dawning from well-defined tertiary structure, the active regions of enzymatic proteins exist as specifically tailored electrostatic microenvironments capable of facilitating chemical interaction. The specific influence these charge distributions have on ligand binding dynamics, and their impact on specificity, reactivity, and biological functionality, have yet to be fully understood. A quantitative determination of these intrinsic fields would offer insight towards the mechanistic aspects of protein functionality. This work seeks to investigate the internal molecular electric fields that are present at the oxygen binding site of myoglobin.
Experiments are performed at 1 K on samples located within a glassy matrix, using the high-resolution technique spectral hole-burning. The internal electric field distributions can be explored by implementing a unique mathematical treatment for analyzing the effect that externally applied electric fields have on the spectral hole profiles. Precise control of the light field, the temperature, and the externally applied electric field at the site of the sample is crucial.
Experimentally, the functionality of custom cryogenic temperature confocal scanning microscope was extended to allow for collection of imaging and spectral data with the ability to modulate the polarization of the light at the sample. Operation of the instrumentation was integrated into a platform allowing for seamless execution of input commands with high temporal inter-instrument resolution for collection of data streams. For the regulated control and cycling of the sample temperature. the thermal characteristics of the research Dewar were theoretically modeled to systematically predict heat flows throughout the system. A high voltage feedthrough for delivering voltages of up to 5000 V to the sample as positioned within the Dewar was developed. The burning of spectral holes with this particular experimental setup is highly repeatable.
The quantum mechanical treatment that is employed during analysis of the experimental data requires the state energies and the transition dipole moments of the porphyrin probe. The configuration interaction, as well as the coupled-cluster approaches, have been investigated for their ability to produce realistic valuations for these calculated quantities as gauged by their ability to accurately reproduce valuations for spectroscopically observable transition energies. A capacitive cell, for the determination of a material’s dielectric permittivity, necessary for defining the magnitude of the externally applied electric field at the sample, was developed and shown to successfully yield permittivity valuations for various media in accordance with those reported the literature, while offering the ability to provide measures for permittivities over the temperature range of 1-300 K
Size-fractionated 234Th in continental shelf waters off New England: Implications for the role of colloids in oceanic trace metal scavenging
Measurements of 234Th (t1/2 = 24.1 days) in dissolved, colloidal, and particulate forms have been made to investigate the role of colloids in reactive metal scavenging in the surface waters of Buzzards Bay, over an annual cycle, and in the shelf and slope waters off New England. At-sea sampling involved prefiltering seawater through 0.2 μm filters followed by cross-flow filtration using a 10,000 nominal molecular weight filter to collect colloidal (10,000 NMW-0.2 μm) and dissolved (\u3c10,000 NMW) phases. Total 234Th activities increase with distance from shore, indicative of enhanced scavenging in the particle-rich nearshore waters. Clearly seen in Buzzards Bay are seasonal changes in total 234Th, with activities ranging from ≈0.7 dpm I−1 in the winter, preceeding a phytoplankton bloom, to ≈0.2 dpm I−1 in the summer. Throughout the annual cycle, 2–16% of total 234Th is colloidal, 22–40% is dissolved, and 45–75% is particulate. In the offshore waters, ≈1% of total 234Th is colloidal, 2–6% is particulate, and 93–98% is dissolved. The 234Th size-distribution exhibits a systematic increase in the association of 234Th with particulate and, to a lesser extent, colloidal matter with increasing suspended particle concentration (Cp). Moreover, a first-order prediction of the fractionation of 234Th between the various size classes is demonstrated using measured solid-solution partition coefficients. Box model calculations indicate a mean residence time of colloidal 234Th with respect to aggregation into particles of 0.3 days in Buzzards Bay, which compares with 2 days for dissolved and 4 days for particulate 234Th. In the offshore surface waters, colloidal and particulate 234Th residence times are ≈0.5 days and 2–3 days respectively, compared with 30–85 days for the dissolved phase. The short and relatively invariant residence time of colloidal 234Th suggests that colloidal aggregation may not be rate-limiting in controlling the scavenging of thorium and, by analogy, other particle-reactive trace metals. An implication of these results is that colloidal 234Th may be tracing a reactive intermediate in the bacterially mediated decomposition of large, rapidly-sinking biogenic aggregates. Using the size-fractionated 234Th data, we demonstrate that Kd values for thorium are invariant with Cp and that scavenging rate constants exhibit a first-order dependence on Cp. Thus, “particle-concentration effects” are negligible for oceanic waters (Cp ≈0.01–1 mg I−1)
Gallium nanoparticles facilitate phagosome maturation and inhibit growth of virulent Mycobacterium tuberculosis in macrophages.
New treatments and novel drugs are required to counter the growing problem of drug-resistant strains of Mycobacterium tuberculosis (M.tb). Our approach against drug resistant M.tb, as well as other intracellular pathogens, is by targeted drug delivery using nanoformulations of drugs already in use, as well as drugs in development. Among the latter are gallium (III) (Ga)-based compounds. In the current work, six different types of Ga and rifampin nanoparticles were prepared in such a way as to enhance targeting of M.tb infected-macrophages. They were then tested for their ability to inhibit growth of a fully pathogenic strain (H37Rv) or a non-pathogenic strain (H37Ra) of M.tb. Encapsulating Ga in folate- or mannose-conjugated block copolymers provided sustained Ga release for 15 days and significantly inhibited M.tb growth in human monocyte-derived macrophages. Nanoformulations with dendrimers encapsulating Ga or rifampin also showed promising anti-tuberculous activity. The nanoparticles co-localized with M.tb containing phagosomes, as measured by detection of mature cathepsin D (34 kDa, lysosomal hydrogenase). They also promoted maturation of the phagosome, which would be expected to increase macrophage-mediated killing of the organism. Delivery of Ga or rifampin in the form of nanoparticles to macrophages offers a promising approach for the development of new therapeutic anti-tuberculous drugs
Agouti C57BL/6N embryonic stem cells for mouse genetic resources.
We report the characterization of a highly germline competent C57BL/6N mouse embryonic stem cell line, JM8. To simplify breeding schemes, the dominant agouti coat color gene was restored in JM8 cells by targeted repair of the C57BL/6 nonagouti mutation. These cells provide a robust foundation for large-scale mouse knockout programs that aim to provide a public resource of targeted mutations in the C57BL/6 genetic background
Recommended from our members
Selective incorporation of proteinaceous over nonproteinaceous cationic amino acids in model prebiotic oligomerization reactions.
Numerous long-standing questions in origins-of-life research center on the history of biopolymers. For example, how and why did nature select the polypeptide backbone and proteinaceous side chains? Depsipeptides, containing both ester and amide linkages, have been proposed as ancestors of polypeptides. In this paper, we investigate cationic depsipeptides that form under mild dry-down reactions. We compare the oligomerization of various cationic amino acids, including the cationic proteinaceous amino acids (lysine, Lys; arginine, Arg; and histidine, His), along with nonproteinaceous analogs of Lys harboring fewer methylene groups in their side chains. These analogs, which have been discussed as potential prebiotic alternatives to Lys, are ornithine, 2,4-diaminobutyric acid, and 2,3-diaminopropionic acid (Orn, Dab, and Dpr). We observe that the proteinaceous amino acids condense more extensively than these nonproteinaceous amino acids. Orn and Dab readily cyclize into lactams, while Dab and Dpr condense less efficiently. Furthermore, the proteinaceous amino acids exhibit more selective oligomerization through their α-amines relative to their side-chain groups. This selectivity results in predominantly linear depsipeptides in which the amino acids are α-amine-linked, analogous to today's proteins. These results suggest a chemical basis for the selection of Lys, Arg, and His over other cationic amino acids for incorporation into proto-proteins on the early Earth. Given that electrostatics are key elements of protein-RNA and protein-DNA interactions in extant life, we hypothesize that cationic side chains incorporated into proto-peptides, as reported in this study, served in a variety of functions with ancestral nucleic acid polymers in the early stages of life
Cross-shelf mixing and mid-shelf front dynamics in the Mid-Atlantic Bight evaluated using the radium quartet
Mid-shelf fronts (MSFs) are thought to be ubiquitous in shelf areas. However, their dynamical role in cross-shelf mixing has yet to be fully characterized. In January, February, and April of 2007, radium isotopes (223Ra, t1/2 = 11 d; 224Ra, t1/2 = 3.7 d; 226Ra, t1/2 = 1600 yr; 228Ra, t1/2 = 5.7 yr) were measured along a transect in the Mid-Atlantic Bight to constrain mixing rates at and around a MSF. Cross-shelf eddy diffusivities (Kx) were determined from 223Ra and 224Ra distributions using a variable-depth model. Two key assumptions – minimal advection and negligible benthic radium input – involving the use of 223Ra and 224Ra as tracers of mixing were quantitatively evaluated in order to assess the accuracy of the Kx estimates. Eddy diffusivities over the three-month sampling period range from 0.1 ± 0.05 – 1.6 ± 0.5 × 102 m2 s–1 for 223Ra and from 1.7 ± 0.4 – 2.2 ± 0.6 × 102 m2 s–1 for 224Ra. The temporal variability in Kx is low in comparison to the uncertainty of the derived values, indicating that eddy diffusivity in this area is relatively constant throughout the sampling period. Observations in the Mid-Atlantic Bight differ from theoretical data corresponding to the tidal dispersion frontogenesis model, suggesting that a different mechanism is responsible for MSF formation. Variability in supported 223Ra and 228Ra observed near the front indicates that cross-shelf mixing may be inhibited by MSFs. Conversely, along-shelf transport is enhanced by the front\u27s presence. These results indicate that the equatorward jet associated with the front is capable of effectively transporting dissolved chemicals over hundreds of kilometers
Changes in the relationship between self-reference and emotional valence as a function of dysphoria
The self-positivity bias is found to be an aspect of normal cognitive function. Changes in this bias are usually associated with changes in emotional states, such as dysphoria or depression. The aim of the present study was to clarify the role of emotional valence within self-referential processing. By asking non-dysphoric and dysphoric individuals to rate separately the emotional and self-referential content of a set of 240 words, it was possible to identify the differences in the relationship between self-reference and emotional valence, which are associated with dysphoria. The results support the existence of the self-positivity bias in non-dysphoric individuals. More interestingly, dysphoric individuals were able to accurately identify the emotional content of the word stimuli. They failed, however, to associate this emotional valence with self-reference. These findings are discussed in terms of attributional self-biases and their consequences for cognition
Platelet-rich plasma and other cellular strategies in orthopedic surgery
The use of biologics in the treatment of musculoskeletal disease has become increasingly more common as research studies continue to provide further elucidation of their mechanisms in healing. Platelet-rich plasma, patches, growth factors, and stem cells are among the many biologics under active investigation and have varying levels of success in augmenting surgical or nonoperative interventions. However, the limitations of these treatments exist, and clear guidelines for their indications and application have yet to be established. Well-designed clinical trials will help determine the appropriate future use of biologics to ensure consistent outcomes
Content analysis of promotional material for asthma-related products and therapies on Instagram
BACKGROUND: Increasingly, social media is a source for information about health and disease self-management. We conducted a content analysis of promotional asthma-related posts on Instagram to understand whether promoted products and services are consistent with the recommendations found in the Global Initiative for Asthma (GINA) 2019 guidelines.
METHODS: We collected every Instagram post incorporating a common, asthma-related hashtag between September 29, 2019 and October 5, 2019. Of these 2936 collected posts, we analyzed a random sample of 266, of which, 211 met our inclusion criteria. Using an inductive, qualitative approach, we categorized the promotional posts and compared each post\u27s content with the recommendations contained in the 2019 GINA guidelines. Posts were categorized as consistent with GINA if the content was supported by the GINA guidelines. Posts that promoted content that was not recommended by or was unrelated to the guidelines were categorized as not supported by GINA .
RESULTS: Of 211 posts, 89 (42.2%) were promotional in nature. Of these, a total of 29 (32.6%) were categorized as being consistent with GINA guidelines. The majority of posts were not supported by the guidelines. Forty-one (46.1%) posts promoted content that was not recommended by the current guidelines. Nineteen (21.3%) posts promoted content that was unrelated to the guidelines. The majority of unsupported content promoted non-pharmacological therapies (n = 39, 65%) to manage asthma, such as black seed oil, salt-room therapy, or cupping.
CONCLUSIONS: The majority of Instagram posts in our sample promoted products or services that were not supported by GINA guidelines. These findings suggest a need for providers to discuss online health information with patients and highlight an opportunity for providers and social media companies to promote evidence-based asthma treatments and self-management advice online
Estimates of Micro-, Nano-, and Picoplankton Contributions to Particle Export in the Northeast Pacific
The contributions of micro-, nano-, and picoplankton to particle export were estimated from measurements of size-fractionated particulate 234Th, organic carbon, and phytoplankton indicator pigments obtained during five cruises between 2010 and 2012 along Line P in the subarctic northeast Pacific Ocean. Sinking fluxes of particulate organic carbon (POC) and indicator pigments were calculated from 234Th–238U disequilibria and, during two cruises, measured by sediment trap at Ocean Station Papa. POC fluxes at 100 m ranged from 0.65–7.95 mmol m−2 d−1, similar in magnitude to previous results at Line P. Microplankton pigments dominate indicator pigment fluxes (averaging 69 ± 19% of total pigment flux), while nanoplankton pigments comprised the majority of pigment standing stocks (averaging 64 ± 23% of total pigment standing stock). Indicator pigment loss rates (the ratio of pigment export flux to pigment standing stock) point to preferential export of larger microplankton relative to smaller nano- and picoplankton. However, indicator pigments do not quantitatively trace particle export resulting from zooplankton grazing, which may be an important pathway for the export of small phytoplankton. These results have important implications for understanding the magnitude and mechanisms controlling the biological pump at Line P in particular, and more generally in oligotrophic gyres and high-nutrient, low-chlorophyll regions where small phytoplankton represent a major component of the autotrophic community
- …