90 research outputs found
Sputum Glucose and Glycemic Control in Cystic Fibrosis-Related Diabetes: A Cross-Sectional Study
Cystic fibrosis-related diabetes affects up to half of cystic fibrosis patients and is associated with increased mortality and more frequent pulmonary exacerbations. However, it is unclear to what degree good glycemic control might mitigate these risks and clinical outcomes have not previously been studied in relation to glucose from the lower airways, the site of infection and CF disease progression. We initially hypothesized that diabetic cystic fibrosis patients with glycosylated hemoglobin (HbA_(1c)) > 6.5% have worse pulmonary function, longer and more frequent exacerbations and also higher sputum glucose levels than patients with HbA_(1c) ≤ 6.5% or cystic fibrosis patients without diabetes. To test this, we analyzed spontaneously expectorated sputum samples from 88 cystic fibrosis patients. The median sputum glucose concentration was 0.70 mM (mean, 4.75 mM; range, 0-64.6 mM). Sputum glucose was not correlated with age, sex, body mass index, diabetes diagnosis, glycemic control, exacerbation frequency or length, or pulmonary function. Surprisingly, sputum glucose was highest in subjects with normal glucose tolerance, suggesting the dynamics of glycemic control, sputum glucose and pulmonary infections are more complex than previously thought. Two-year mean HbA_(1c) was positively correlated with the length of exacerbation admission (p 6.5% were hospitalized on average 6 days longer than those with HbA_(1c) ≤ 6.5% (p < 0.01). Current clinical care guidelines for cystic fibrosis-related diabetes target HbA_(1c) ≤ 7% to limit long-term microvascular damage, but more stringent glycemic control (HbA_(1c) ≤ 6.5%) may further reduce the short-term pulmonary complications
Further evidence for fungivory in the Lower Devonian (Lochkovian) of the Welsh Borderland, UK
The recent demonstrations that widespread mid-Palaeozoic Prototaxites and other nematophytes had fungal affinities indicate that terrestrial fungi were important elements in carbon cycling in the Early Devonian. Here, we provide evidence for their participation in the recycling of nutrients by early terrestrial invertebrates. Evidence is in the form of coprolites, both those associated with nematophytes or containing their fragmentary remains. Cylindrical coprolites consistently associated with fungal mats are placed in a new ichnospecies, Bacillafaex myceliorum. Their contents are granular to amorphous, suggestive of complete digestion of the ingested hyphae, with the inference of possession of chitinases in the digestive tracts of the consumers. A further single example comprises a cluster of cylindrical bodies attached to the lower surface of a Nematothallus fragment. Here, homogenisation was less complete, with traces of hyphae remaining. Terrestrial animal fossils have not been found at the locality, but scorpions, pseudoscorpions, Opiliones, mites, centipedes (carnivores) and millipedes, and Collembola (detritivores) have been recorded from the slightly younger Rhynie cherts. Studies of fungivory in extant arthropods have concentrated on Collembola and, to a lesser extent, mites, but their faecal pellets are much smaller than the fossil examples. Millipedes, based on body size and faeces of extant forms, are considered more realistic producers, but little is known about fungal feeding in these animals. Regardless of the affinities of the producers, the diversity in morphology, sizes, aggregations, and composition of nematophyte-containing examples suggests that fungivory was an important component of carbon cycling in early terrestrial ecosystems
Anatomically preserved Silurian 'nematophytes' from the Welsh Borderland (UK)
Stratified charcoalified fragments of thalloid organisms with tripartite tissue construction have been isolated from the basal member of the Upper Silurian (upper Ludlow) Downton Castle Sandstone Formation, exposed near Ludlow, Shropshire (England) and are considered to have had fungal affinity. They are divided into two major groups. The more novel of these is characterized by a superficial cortex separated from a basal layer of interweaving hyphae by an intermediate zone of compressed indeterminate tissue and members are placed in a new taxon, Tristratothallus ludfordensis. In the second, the intermediate zone comprises hyphae arranged at right angles to the cortex (termed palisade). Some members resemble the tissue construction of Nematothallus described from the Lower Devonian (Lochkovian) of the Welsh Borderland and considered to belong to fungi, some of which were lichenized. A further type, which shows remains of polysporic asci, is thought to represent a fragment of an apothecium (a disc-shaped ascoma of an ascomycete) of a pezizomycete and is the earliest such record. Yet others are characterized by a perforate cortex with occasional protruding hyphae, tissue construction of which was also recorded in the Lower Devonian of the Welsh Borderland and considered to display fungal characteristics. Coalified ‘black patches’ are common on bedding surfaces throughout the latest Silurian and Early Devonian and frequently are associated with basal embryophytes and tracheophytes. Those reported here are the oldest known with three-dimensional organization, studied via scanning electron microscopy (SEM) and attributed to fungi, and include some ascomycetes. Similar encrustations occur in even earlier rocks and may have been important constituents of the cryptogamic ground cover, which is postulated to have preceded higher plant life on land
Provenance and age of bacteria-like structures on mid-Palaeozoic plant fossils
Structures, termed microbioids, comforming to bacteria in size and shape (e.g. rods, spheres, chains and clusters of spheres) have been observed by field emission scanning electron microscopy (FE-SEM) on coalified Silurian and Lower Devonian spores, sporangia, cuticles and coprolites. Some were sectioned for transmission electron microscopy. The elemental composition of both microbioids and ‘substrates’ was investigated using a X-ray microanalysis system. These analyses combined with comparative studies on recent bacteria and cyanobacteria were undertaken to evaluate the biogenicity, nature and age of the microbioids. Spheres with a Si signature (0.03–0.5 μm diameter) and assumed composed of silica are interpreted as artefacts produced abiotically during the extraction procedures. A similar origin is proposed for hollow spheres that are composed of CaF2. These occur singly, in short chains simulating filaments, and in clusters. Considerable differences in size (0.2–2.0 μm diameter) and appearance relate to local variation in the chemical environment during extraction. Spheres (0.2–1.5 μm diameter), that lack a mineral signature, with a framboidal surface ornament and occur within sporangia are identified as by-products of spore development. A biotic origin is also postulated for C-containing rod-shaped structures (>3.1 μm long, <1.4 μm wide), some with collapsed surfaces, although comparisons with living bacteria indicate recent contamination. More elongate rod-shaped microbioids (<8.6 μm long, 1.2 μm wide) have been identified as detrital rutile crystals (TiO2). Minute naviculate structures (<2.2 μm long) resembling diatoms are of unknown origin but are probably composed of thorium hydroxide. Unmineralized filaments of cyanobacterial morphology are recent contaminants. Some of the sporangia and spore masses are partially covered by associations of fragmented sheets, interconnecting strands, rods and spheres that are interpreted as dehydrated biofilms. Being unmineralized they are probably also of recent origin, although they might have survived wild-fire along with the charcoalified mesofossils. Many of the structures illustrated here were initially identified casually as bacteria on the small fossils extracted for biodiversity studies using well-tried, conventional, palaeobotanical techniques. Our subsequent more detailed analyses have shown how such processes can produce artefacts that are morphological analogues of mineralized bacteria, leave residues that mimic bacterial shapes and, despite some efforts such as storage in dilute HCl to eliminate living bacteria, introduce contamination. They reinforce previous concerns that verification of the biogenicity and syngenicity of bacterial-like objects in ancient Earth and extra-terrestrial rocks should not only rely on size and morphological look-alikes, but must encompass a thorough understanding of fossilization processes and extraction techniques plus, ideally, other measures of biogenicity (e.g. biomarkers) and syngenicity
Trace incorporation of heavy water reveals slow and heterogeneous pathogen growth rates in cystic fibrosis sputum
Effective treatment for chronic infections is undermined by a significant gap in understanding of the physiological state of pathogens at the site of infection. Chronic pulmonary infections are responsible for the morbidity and mortality of millions of immunocompromised individuals worldwide, yet drugs that are successful in laboratory culture are far less effective against pathogen populations persisting in vivo. Laboratory models, upon which preclinical development of new drugs is based, can only replicate host conditions when we understand the metabolic state of the pathogens and the degree of heterogeneity within the population. In this study, we measured the anabolic activity of the pathogen Staphylococcus aureus directly in the sputum of pediatric patients with cystic fibrosis (CF), by combining the high sensitivity of isotope ratio mass spectrometry with a heavy water labeling approach to capture the full range of in situ growth rates. Our results reveal S. aureus generation times with a median of 2.1 d, with extensive growth rate heterogeneity at the single-cell level. These growth rates are far below the detection limit of previous estimates of CF pathogen growth rates, and the rates are slowest in acutely sick patients undergoing pulmonary exacerbations; nevertheless, they are accessible to experimental replication within laboratory models. Treatment regimens that include specific antibiotics (vancomycin, piperacillin/tazobactam, tobramycin) further appear to correlate with slow growth of S. aureus on average, but follow-up longitudinal studies must be performed to determine whether this effect holds for individual patients
Piecing together the eophytes - a new group of ancient plants containing cryptospores
The earliest evidence for land plants comes from dispersed cryptospores from the Ordovician, which dominated assemblages for 60 million years. Direct evidence of their parent plants comes from minute fossils in Welsh Borderland Upper Silurian to Lower Devonian rocks. We recognize a group that had forking, striated axes with rare stomata terminating in valvate sporangia containing permanent cryptospores, but their anatomy was unknown especially regarding conducting tissues.
Charcoalified fossils extracted from the rock using HF were selected from macerates and observed using scanning electron microscopy. Promising examples were split for further examination and compared with electron micrographs of the anatomy of extant bryophytes.
Fertile fossil axes possess central elongate cells with thick walls bearing globules, occasional strands and plasmodesmata-sized pores. The anatomy of these cells best matches desiccation-tolerant food-conducting cells (leptoids) of bryophytes. Together with thick-walled epidermal cells and extremely small size, these features suggest that these plants were poikilohydric.
Our new data on conducting cells confirms a combination of characters that distinguish the permanent cryptospore-producers from bryophytes and tracheophytes. We therefore propose the erection of a new group, here named the Eophytidae (eophytes)
Earliest record of transfer cells in Lower Devonian plants
Key sources of information on the nature of early terrestrial ecosystems are the fossilized remains of plants and associated organic encrustations, which are interpreted as either biofilms, biological soil crusts or lichens. The hypothesis that some of these encrustations might be the remains of the thalloid gametophytes of embryophytes provided the stimulus for this investigation.
Fossils preserved in charcoal were extracted from Devonian Period (Lochkovian Stage, c. 410–419 Myr old) sediments at a geological site in Shropshire (UK). Scanning electron micrographs (SEMs) of the fossils were compared with new and published SEMs of extant bryophytes and tracheophytes, respectively. One specimen was further prepared and imaged by transmission electron microscopy.
Fossils of thalloid morphology were composed almost entirely of cells with labyrinthine ingrowths; these also were present in fossils of axial morphology where they were associated with putative food-conducting cells. Comparison with modern embryophytes demonstrates that these distinctive cells are transfer cells (TCs).
Our fossils provide by far the earliest geological evidence of TCs. They also show that some organic encrustations are the remains of thalloid land plants and that these are possibly part of the life cycle of a newly recognized group of plants called the eophyte
Exposing the Three-Dimensional Biogeography and Metabolic States of Pathogens in Cystic Fibrosis Sputum via Hydrogel Embedding, Clearing, and rRNA Labeling
Physiological resistance to antibiotics confounds the treatment of many chronic bacterial infections, motivating researchers to identify novel therapeutic approaches. To do this effectively, an understanding of how microbes survive in vivo is needed. Though much can be inferred from bulk approaches to characterizing complex environments, essential information can be lost if spatial organization is not preserved. Here, we introduce a tissue-clearing technique, termed MiPACT, designed to retain and visualize bacteria with associated proteins and nucleic acids in situ on various spatial scales. By coupling MiPACT with hybridization chain reaction (HCR) to detect rRNA in sputum samples from cystic fibrosis (CF) patients, we demonstrate its ability to survey thousands of bacteria (or bacterial aggregates) over millimeter scales and quantify aggregation of individual species in polymicrobial communities. By analyzing aggregation patterns of four prominent CF pathogens, Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus sp., and Achromobacter xylosoxidans, we demonstrate a spectrum of aggregation states: from mostly single cells (A. xylosoxidans), to medium-sized clusters (S. aureus), to a mixture of single cells and large aggregates (P. aeruginosa and Streptococcus sp.). Furthermore, MiPACT-HCR revealed an intimate interaction between Streptococcus sp. and specific host cells. Lastly, by comparing standard rRNA fluorescence in situ hybridization signals to those from HCR, we found that different populations of S. aureus and A. xylosoxidans grow slowly overall yet exhibit growth rate heterogeneity over hundreds of microns. These results demonstrate the utility of MiPACT-HCR to directly capture the spatial organization and metabolic activity of bacteria in complex systems, such as human sputum
New counselor educators’ scholarly productivity: Supportive and discouraging environments
Eighteen new counselor educators shared in-depth reports of their efforts to establish an early record of scholarly productivity, critical to their success in academe. Analyses of their experiences using consensual qualitative research methods revealed components of both highly positive and highly discouraging program environments and their impact on participants’ research goals
Fertile Prototaxites taiti: a basal ascomycete with inoperculate, polysporous asci lacking croziers
The affinities of Prototaxites have been debated ever since its fossils, some attaining tree-trunk proportions, were discovered in Canadian Lower Devonian rocks in 1859. Putative assignations include conifers, red and brown algae, liverworts and fungi (some lichenised). Detailed anatomical investigation led to the reconstruction of the type species, P. logani, as a giant sporophore (basidioma) of an agaricomycete (= holobasidiomycete), but evidence for its reproduction remained elusive. Tissues associated with P. taiti in the Rhynie chert plus charcoalified fragments from southern Britain are investigated here to describe the reproductive characters and hence affinities of Prototaxites. Thin sections and peels (Pragian Rhynie chert, Aberdeenshire) were examined using light and confocal microscopy; PÅ™Ãdolà and Lochkovian charcoalified samples (Welsh Borderland) were liberated from the rock and examined with scanning electron microscopy. Prototaxites taiti possessed a superficial hymenium comprising an epihymenial layer, delicate septate paraphyses, inoperculate polysporic asci lacking croziers and a subhymenial layer composed predominantly of thin-walled hyphae and occasional larger hyphae. Prototaxites taiti combines features of extant Taphrinomycotina (Neolectomycetes lacking croziers) and Pezizomycotina (epihymenial layer secreted by paraphyses) but is not an ancestor of the latter. Brief consideration is given to its nutrition and potential position in the phylogeny of the Ascomycota
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