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At least 37 records · Page 2

A pollen-specific novel calmodulin-binding protein with tetratricopeptide repeats

Calcium is essential for pollen germination and pollen tube growth. A large body of information has established a link between elevation of cytosolic Ca(2+) at the pollen tube tip and its growth. Since the action of Ca(2+) is primarily mediated by Ca(2+)-binding proteins such as calmodulin (CaM), identification of CaM-binding proteins in pollen should provide insights into the mechanisms by which Ca(2+) regulates pollen germination and tube growth. In this study, a CaM-binding protein from maize pollen (maize pollen calmodulin-binding protein, MPCBP) was isolated in a protein-protein interaction-based screening using (35)S-labeled CaM as a probe. MPCBP has a molecular mass of about 72 kDa and contains three tetratricopeptide repeats (TPR) suggesting that it is a member of the TPR family of proteins. MPCBP protein shares a high sequence identity with two hypothetical TPR-containing proteins from Arabidopsis. Using gel overlay assays and CaM-Sepharose binding, we show that the bacterially expressed MPCBP binds to bovine CaM and three CaM isoforms from Arabidopsis in a Ca(2+)-dependent manner. To map the CaM-binding domain several truncated versions of the MPCBP were expressed in bacteria and tested for their ability to bind CaM. Based on these studies, the CaM-binding domain was mapped to an 18-amino acid stretch between the first and second TPR regions. Gel and fluorescence shift assays performed with CaM and a CaM-binding synthetic peptide further confirmed MPCBP binding to CaM. Western, Northern, and reverse transcriptase-polymerase chain reaction analysis have shown that MPCBP expression is specific to pollen. MPCBP was detected in both soluble and microsomal proteins. Immunoblots showed the presence of MPCBP in mature and germinating pollen. Pollen-specific expression of MPCBP, its CaM-binding properties, and the presence of TPR motifs suggest a role for this protein in Ca(2+)-regulated events during pollen germination and growth.

NASA Discipline Plant Biology↗

Identification of a new EF-hand superfamily member from Trypanosoma brucei

We identified several open reading frames between the regions encoding calmodulin and ubiquitin-EP52/1 in the genome of Trypanosoma brucei. One of these, EFH5, encodes a protein 192 amino acids long. The EFH5 transcript is present in poly(A)+ mRNA and is present at similar levels in the mammalian bloodstream form and the insect procyclic form. EFH5 contains four EF-hand homolog domains, two of which are inferred to bind Ca2+ ions. We expressed EFH5 as a fusion protein in Escherichia coli and demonstrated calcium-binding activity of the fusion protein using the 45Ca-overlay technique. The function of EFH5 remains unknown; however, as the fourth EF-hand homolog identified in trypanosomes, it attests to the broad range of functions assumed by calcium functioning as a second messenger. EFH5, which is most closely related to LAV1-2 from Physarum, represents a distinct subfamily among the EF-hand-containing proteins.

Non-NASA Center↗

Testing putative hemichordate homologues of the chordate dorsal nervous system and endostyle: expression of NK2.1 (TTF-1) in the acorn worm Ptychodera flava (Hemichordata, Ptychoderidae)

Recent phylogenetic investigations have confirmed that hemichordates and echinoderms are sister taxa. However, hemichordates share several cardinal characterstics with chordates and are thus an important taxon for testing hypotheses of homology between key chordate characters and their putative hemichordate antecedents. The chordate dorsal nervous system (DNS) and endostyle are intriguing characters because both hemichordate larval and adult structures have been hypothesized as homologues. This study attempts to test these purported homologies through examination of the expression pattem of a Ptychodera flava NK2 gene, PfNK2.1, because this gene is expressed both in the DNS and endostyle/thyroid in a wide range of chordate taxa. We found that PfNK2.1 is expressed in both neuronal and pharyngeal structures, but its expression pattem is broken up into distinct embryonic and juvenile phases. During embryogenesis, PfNK2.1 is expressed in the apical ectoderm, with transcripts later detected in presumable neuronal structures, including the apical organ and ciliated feeding band. In the developing juvenile we detected PfNK2.1 signal throughout the pharynx, including the stomochord, and later in the hindgut. We conclude that the similar utilization of NK2.1 in apical organ development and chordate DNS is probably due to a more general role for NK2.1 in neurogenesis and that hemichordates do not possess a homologue of the chordate DNS. In addition, we conclude that P. flava most likely does not possess a true endostyle; rather during the evolution of the endostyle NK2.1 was recruited from its more general role in pharynx development.

NASA Discipline Evolutionary Biology↗

Id expression in amphioxus and lamprey highlights the role of gene cooption during neural crest evolution

Neural crest cells are unique to vertebrates and generate many of the adult structures that differentiate them from their closest invertebrate relatives, the cephalochordates. Id genes are robust markers of neural crest cells at all stages of development. We compared Id gene expression in amphioxus and lamprey to ask if cephalochordates deploy Id genes at the neural plate border and dorsal neural tube in a manner similar to vertebrates. Furthermore, we examined whether Id expression in these cells is a basal vertebrate trait or a derived feature of gnathostomes. We found that while expression of Id genes in the mesoderm and endoderm is conserved between amphioxus and vertebrates, expression in the lateral neural plate border and dorsal neural tube is a vertebrate novelty. Furthermore, expression of lamprey Id implies that recruitment of Id genes to these cells occurred very early in the vertebrate lineage. Based on expression in amphioxus we postulate that Id cooption conferred sensory cell progenitor-like properties upon the lateral neurectoderm, and pharyngeal mesoderm-like properties upon cranial neural crest. Amphioxus Id expression is also consistent with homology between the anterior neurectoderm of amphioxus and the presumptive placodal ectoderm of vertebrates. These observations support the idea that neural crest evolution was driven in large part by cooption of multipurpose transcriptional regulators from other tissues and cell types.

NASA Program Fundamental Space Biology↗

Analysis and expression of the alpha-expansin and beta-expansin gene families in maize

Expansins comprise a multigene family of proteins in maize (Zea mays). We isolated and characterized 13 different maize expansin cDNAs, five of which are alpha-expansins and eight of which are beta-expansins. This paper presents an analysis of these 13 expansins, as well as an expression analysis by northern blotting with materials from young and mature maize plants. Some expansins were expressed in restricted regions, such as the beta-expansins ExpB1 (specifically expressed in maize pollen) and ExpB4 (expressed principally in young husks). Other expansins such as alpha-expansin Exp1 and beta-expansin ExpB2 were expressed in several organs. The expression of yet a third group was not detected in the selected organs and tissues. An analysis of expansin sequences from the maize expressed sequence tag collection is also presented. Our results indicate that expansin genes may have general, overlapping expression in some instances, whereas in other cases the expression may be highly specific and limited to a single organ or cell type. In contrast to the situation in Arabidopsis, beta-expansins in maize seem to be more numerous and more highly expressed than are alpha-expansins. The results support the concept that beta-expansins multiplied and evolved special functions in the grasses.

NASA Program Fundamental Space Biology↗

A low, adaptive dose of gamma-rays reduced the number and altered the spectrum of S1- mutants in human-hamster hybrid AL cells

We examined the effects of a low, adaptive dose of 137Cs-gamma-irradiation (0.04 Gy) on the number and kinds of mutants induced in AL human-hamster hybrid cells by a later challenge dose of 4 Gy. The yield of S1- mutants was significantly less (by 53%) after exposure to both the adaptive and challenge doses compared to the challenge dose alone. The yield of hprt- mutants was similarly decreased. Incubation with cycloheximide (CX) or 3-aminobenzamide largely negated the decrease in mutant yield. The adaptive dose did not perturb the cell cycle, was not cytotoxic, and did not of itself increase the mutant yield above background. The adaptive dose did, however, alter the spectrum of S1- mutants from populations exposed only to the adaptive dose, as well as affecting the spectrum of S1- mutants generated by the challenge dose. The major change in both cases was a significant increase in the proportion of complex mutations compared to small mutations and simple deletions.

Non-NASA Center↗

Non-random distribution of DNA double-strand breaks induced by particle irradiation

Induction of DNA double-strand breaks (dsbs) in mammalian cells is dependent on the spatial distribution of energy deposition from the ionizing radiation. For high LET particle radiations the primary ionization sites occur in a correlated manner along the track of the particles, while for X-rays these sites are much more randomly distributed throughout the volume of the cell. It can therefore be expected that the distribution of dsbs linearly along the DNA molecule also varies with the type of radiation and the ionization density. Using pulsed-field gel and conventional gel techniques, we measured the size distribution of DNA molecules from irradiated human fibroblasts in the total range of 0.1 kbp-10 Mbp for X-rays and high LET particles (N ions, 97 keV/microns and Fe ions, 150 keV/microns). On a mega base pair scale we applied conventional pulsed-field gel electrophoresis techniques such as measurement of the fraction of DNA released from the well (FAR) and measurement of breakage within a specific NotI restriction fragment (hybridization assay). The induction rate for widely spaced breaks was found to decrease with LET. However, when the entire distribution of radiation-induced fragments was analysed, we detected an excess of fragments with sizes below about 200 kbp for the particles compared with X-irradiation. X-rays are thus more effective than high LET radiations in producing large DNA fragments but less effective in the production of smaller fragments. We determined the total induction rate of dsbs for the three radiations based on a quantitative analysis of all the measured radiation-induced fragments and found that the high LET particles were more efficient than X-rays at inducing dsbs, indicating an increasing total efficiency with LET. Conventional assays that are based only on the measurement of large fragments are therefore misleading when determining total dsb induction rates of high LET particles. The possible biological significance of this non-randomness for dsb induction is discussed.

NASA Discipline Radiation Health↗

Multiple melanocortin receptors are expressed in bone cells

Melanocortin receptors belong to the seven transmembrane domain, G-protein coupled family of receptors. There are five members of this receptor family labeled MC1R-MC5R. These receptors are activated by fragments derived from a larger molecule, proopiomelanocortin (POMC) and include ACTH, alpha beta and gamma-MSH and beta-endorphin. Because of in vitro and in vivo data suggesting direct effects of these POMC molecules on bone and bone turnover, we examined bone and bone derived cells for the presence of the various members of the melanocortin receptor family. We report that the five known melanocortin receptors are expressed to varying degrees in osteoblast-like and osteoclastic cells. POMC fragments increased proliferation and expression of a variety of genes in osteoblastic cells. Furthermore, POMC mRNA was detected in osteoclastic cells. These data demonstrate that POMC-derived peptide hormones acting through high affinity melanocortin receptors have specific effects on bone cells. Thus, in addition to the indirect effects of POMC-derived hormones on bone turnover through their modulation of steroid hormone secretion, POMC fragments may have direct and specific effects on bone cell subpopulations.

NASA Discipline Musculoskeletal↗

Sequence, molecular properties, and chromosomal mapping of mouse lumican

PURPOSE. Lumican is a major proteoglycan of vertebrate cornea. This study characterizes mouse lumican, its molecular form, cDNA sequence, and chromosomal localization. METHODS. Lumican sequence was determined from cDNA clones selected from a mouse corneal cDNA expression library using a bovine lumican cDNA probe. Tissue expression and size of lumican mRNA were determined using Northern hybridization. Glycosidase digestion followed by Western blot analysis provided characterization of molecular properties of purified mouse corneal lumican. Chromosomal mapping of the lumican gene (Lcn) used Southern hybridization of a panel of genomic DNAs from an interspecific murine backcross. RESULTS. Mouse lumican is a 338-amino acid protein with high-sequence identity to bovine and chicken lumican proteins. The N-terminus of the lumican protein contains consensus sequences for tyrosine sulfation. A 1.9-kb lumican mRNA is present in cornea and several other tissues. Antibody against bovine lumican reacted with recombinant mouse lumican expressed in Escherichia coli and also detected high molecular weight proteoglycans in extracts of mouse cornea. Keratanase digestion of corneal proteoglycans released lumican protein, demonstrating the presence of sulfated keratan sulfate chains on mouse corneal lumican in vivo. The lumican gene (Lcn) was mapped to the distal region of mouse chromosome 10. The Lcn map site is in the region of a previously identified developmental mutant, eye blebs, affecting corneal morphology. CONCLUSIONS. This study demonstrates sulfated keratan sulfate proteoglycan in mouse cornea and describes the tools (antibodies and cDNA) necessary to investigate the functional role of this important corneal molecule using naturally occurring and induced mutants of the murine lumican gene.

NASA Discipline Cell Biology↗

Time-Integrated Neutrino Source Searches with 10 Years of IceCube Data

This Letter presents the results from pointlike neutrino source searches using ten years of IceCube data collected between April 6, 2008 and July 10, 2018. We evaluate the significance of an astrophysical signal from a pointlike source looking for an excess of clustered neutrino events with energies typically above ~1 TeV among the background of atmospheric muons and neutrinos. We perform a full-sky scan, a search within a selected source catalog, a catalog population study, and three stacked Galactic catalog searches. The most significant point in the northern hemisphere from scanning the sky is coincident with the Seyfert II galaxy NGC 1068, which was included in the source catalog search. The excess at the coordinates of NGC 1068 is inconsistent with background expectations at the level of 2.9σ after accounting for statistical trials from the entire catalog. The combination of this result along with excesses observed at the coordinates of three other sources, including TXS 0506+056, suggests that, collectively, correlations with sources in the northern catalog are inconsistent with background at 3.3σ significance. The southern catalog is consistent with background. Finally these results, all based on searches for a cumulative neutrino signal integrated over the 10 years of available data, motivate further study of these and similar sources, including time-dependent analyses, multimessenger correlations, and the possibility of stronger evidence with coming upgrades to the detector.

79 ASTRONOMY AND ASTROPHYSICS↗

All-sky Neutrino Point-source Search with IceCube Combined Track and Cascade Data

Despite extensive efforts, discovery of high-energy astrophysical neutrino sources remains elusive. We present an event-level simultaneous maximum likelihood analysis of tracks and cascades using IceCube data collected from 2008 April 6 to 2022 May 23 to search the whole sky for neutrino sources, and using a source catalog, for coincidence of neutrino emission with gamma-ray emission. This is the first time a simultaneous fit of different detection channels is used to conduct a time-integrated all-sky scan with IceCube. Combining all-sky tracks, with superior pointing power and sensitivity in the northern sky, with all-sky cascades, with good energy resolution and sensitivity in the southern sky, we have developed the most sensitive point-source search to date by IceCube that targets the entire sky. The most significant point in the northern sky aligns with NGC 1068, a Seyfert II galaxy, which, from the catalog search, shows a 3.5σ excess over background after accounting for trials. The most significant point in the southern sky does not align with any source in the catalog and is not significant after accounting for trials. A search for the single most significant Gaussian flare at the locations of NGC 1068, PKS 1424+240, and the southern highest-significance point shows results consistent with expectations for steady emission. Notably, this is the first time that a flare shorter than four years has been excluded as being responsible for NGC 1068’s emergence as a neutrino source. Our results show that combining tracks and cascades when conducting neutrino source searches improves sensitivity and can lead to new discoveries.

Abbasi, R. [Loyola University, Chicago, IL (United↗

Characterization of the astrophysical diffuse neutrino flux using starting track events in IceCube

In this article, a measurement of the diffuse astrophysical neutrino spectrum is presented using IceCube data collected from 2011-2022 (10.3 years). We developed novel detection techniques to search for events with a contained vertex and exiting track induced by muon neutrinos undergoing a charged-current interaction. Searching for these starting track events allows us to not only more effectively reject atmospheric muons but also atmospheric neutrino backgrounds in the southern sky, opening a new window to the sub-100 TeV astrophysical neutrino sky. The event selection is constructed using a dynamic starting track veto and machine learning algorithms. We use this data to measure the astrophysical diffuse flux as a single power law flux (SPL) with a best-fit spectral index of γ=2.58$_{-0.09}^{+0.10}$ and per-flavor normalization of $\phi$$_{per-flavor}^{Astro}$=1.68$_{-0.22}^{+0.19}$×10 -18 ×GeV -1 cm -2 s -1 sr -1 (at 100 TeV). The sensitive energy range for this dataset is 3-550 TeV under the SPL assumption. This data was also used to measure the flux under a broken power law, however we did not find any evidence of a low energy cutoff.

79 ASTRONOMY AND ASTROPHYSICS↗

Evidence for Neutrino Emission from X-Ray Bright Seyfert Galaxies in the Southern Hemisphere Using Enhanced Starting Track Events with IceCube

IceCube recently reported the observation of TeV neutrinos from the nearby Seyfert galaxy NGC 1068, and the corresponding neutrino flux is significantly higher than the upper limit implied by observations of GeV–TeV gamma rays. This suggests that neutrinos are produced near the supermassive black hole, where the radiation density is high enough to obscure gamma rays. We use a set of muon neutrinos with interaction vertices inside the detector, which have good sensitivity to sources in the southern sky, from IceCube data recorded between 2011 and 2021. We then search for individual and collective neutrino signals from 14 Seyfert galaxies in the southern sky selected from the Swift Burst Alert Telescope AGN Spectroscopic Survey. Using the correlations between keV X-rays and TeV neutrinos predicted by disk–corona models, and assuming production characteristics similar to NGC 1068, a collective neutrino signal search reveals an excess of $6.{7}_{-3.2}^{+4.0}$ events, which is inconsistent with background expectations at the 3σ level of significance. In this Letter, we present new independent evidence that Seyfert galaxies contribute to the extragalactic flux of high-energy neutrinos.

Seyfert galaxies↗

Time-integrated Southern-sky Neutrino Source Searches with 10 yr of IceCube Starting-track Events at Energies Down to 1 TeV

In the IceCube Neutrino Observatory, a signal of astrophysical neutrinos is obscured by backgrounds from atmospheric neutrinos and muons produced in cosmic-ray interactions. IceCube event selections used to isolate the astrophysical neutrino signal often focus on the morphology of the light patterns recorded by the detector. The analyses presented here use the new IceCube Enhanced Starting Track Event Selection (ESTES), which identifies events likely generated by muon–neutrino interactions within the detector geometry, focusing on neutrino energies of 1–500 TeV with a median angular resolution of 1.4°. Selecting for starting-track events filters out not only the atmospheric-muon background but also the atmospheric-neutrino background in the southern sky. This improves IceCube’s muon–neutrino sensitivity to southern-sky neutrino sources, especially for Galactic sources that are not expected to produce a substantial flux of neutrinos above 100 TeV. In this work, the ESTES sample was applied for the first time to search for astrophysical sources of neutrinos, including a search for diffuse neutrino emission from the Galactic plane. No significant excesses were identified from any of the analyses; however, constraining limits are set on the hadronic emission from TeV gamma-ray Galactic plane objects and models of the diffuse Galactic plane neutrino flux.

Abbasi, R. [Loyola University, Chicago, IL (United↗

Probing the PeV region in the astrophysical neutrino spectrum using 𝜈 𝜇 from the Southern sky

IceCube has observed a diffuse astrophysical neutrino flux over the energy region from a few TeV to a few PeV. At PeV energies, the spectral shape is not yet well measured due to the low statistics of the data. This analysis probes the gap between 1 and 10 PeV by using high-energy downgoing muon neutrinos. Here, to reject the large atmospheric muon background, two complementary techniques are combined. The first technique selects events with high stochasticity to reject atmospheric muon bundles whose stochastic energy losses are smoothed due to high muon multiplicity. The second technique vetoes atmospheric muons with the IceTop surface array. Using 9 yrs of data, we found two neutrino candidate events in the signal region, consistent with expectation from background, each with relatively high signal probabilities. A joint maximum likelihood estimation is performed using this sample and an independent 9.5-yr sample of tracks to measure the neutrino spectrum. A likelihood ratio test is done to compare the single power-law (SPL) vs SPL+cutoff hypothesis; the SPL+cutoff model is not significantly better than the SPL. High-energy astrophysical objects from four source catalogs are also checked around the direction of the two events. No significant coincidence was found.

Abbasi, R. [Loyola University Chicago] (ORCID:0000↗

Observation of Cosmic-Ray Anisotropy in the Southern Hemisphere with 12 yr of Data Collected by the IceCube Neutrino Observatory

We analyzed the 7.92 × 10 11 cosmic-ray-induced muon events collected by the IceCube Neutrino Observatory from 2011 May 13, when the fully constructed experiment started to take data, to 2023 May 12. This data set provides an up-to-date cosmic-ray arrival direction distribution in the Southern Hemisphere with unprecedented statistical accuracy covering more than a full period length of a solar cycle. Improvements in Monte Carlo event simulation and better handling of year-to-year differences in data processing significantly reduce systematic uncertainties below the level of statistical fluctuations compared to the previously published results. We confirm the observation of a change in the angular structure of the cosmic-ray anisotropy between 10 TeV and 1 PeV, more specifically in the 100–300 TeV energy range. For the first time, we analyzed the angular power spectrum at different energies. The observed variations of the power spectra with energy suggest relatively reduced large-scale features at high energy compared to those of medium and small scales. The large volume of data enhances the statistical significance at higher energies, up to the PeV scale, and smaller angular scales, down to approximately 6° compared to previous findings.

79 ASTRONOMY AND ASTROPHYSICS↗

Search for PeV Gamma-Ray Emission from the Southern Hemisphere with 5 Yr of Data from the IceCube Observatory

The measurement of diffuse PeV gamma-ray emission from the Galactic plane would provide information about the energy spectrum and propagation of Galactic cosmic rays, and the detection of a pointlike source of PeV gamma-rays would be strong evidence for a Galactic source capable of accelerating cosmic rays up to at least a few PeV. This paper presents several unbinned maximum-likelihood searches for PeV gamma-rays in the Southern Hemisphere using 5 yr of data from the IceTop air shower surface detector and the in-ice array of the IceCube Observatory. The combination of both detectors takes advantage of the low muon content and deep shower maximum of gamma-ray air showers and provides excellent sensitivity to gamma-rays between ~0.6 and 100 PeV. Our measurements of pointlike and diffuse Galactic emission of PeV gamma-rays are consistent with the background, so we constrain the angle-integrated diffuse gamma-ray flux from the Galactic plane at 2 PeV to 2.61 × 10 -19 cm -2 s -1 TeV -1 at 90% confidence, assuming an E -3 spectrum, and we estimate 90% upper limits on pointlike emission at 2 PeV between 10 -21 and 10 -20 cm -2 s -1 TeV -1 for an E -2 spectrum, depending on decl. Furthermore, we exclude unbroken power-law emission up to 2 PeV for several TeV gamma-ray sources observed by the High Energy Spectroscopic System and calculate upper limits on the energy cutoffs of these sources at 90% confidence. We also find no PeV gamma-rays correlated with neutrinos from IceCube's high-energy starting event sample. Lastly, these are currently the strongest constraints on PeV gamma-ray emission.

79 ASTRONOMY AND ASTROPHYSICS↗

Probing neutrino emission at GeV energies from compact binary mergers with the IceCube Neutrino Observatory

The advent of multimessenger astronomy has allowed for new types of source searches by neutrino detectors. Here, we present the results of the search for 0.5–100 GeV astrophysical neutrinos detected with IceCube and emitted from compact binary mergers detected by the LIGO, Virgo, and KAGRA interferometers from their first run of observation (O1) to the end of the first part of the fourth (O4a). An innovative approach is used to lower the energy threshold to 0.5 GeV and to search for an excess of GeV neutrinos in time coincidence with astrophysical transient events. Furthermore, we use a statistical combination of all observations, a binomial test, to search for a subpopulation of neutrino emitters. No significant excess was found from the studied mergers, with a best post-trial 𝑝-value of 40%, and there is currently no hint of a population of GeV neutrino emitters found in the IceCube data (post-trial 𝑝−value=81%).

Abbasi, R. [Loyola University Chicago]↗