Pulmonary arteries in coelacanths shed light on the vasculature evolution of air-breathing organs in vertebrates.

To date, the presence of pulmonary organs in the fossil record is extremely rare. Among extant vertebrates, lungs are described in actinopterygian polypterids and in all sarcopterygians, including coelacanths and lungfish. However, vasculature of pulmonary arteries has never been accurately identified neither in fossil nor extant coelacanths due to the paucity of fossil preservation of pulmonary organs and limitations of invasive studies in extant specimens. Here we present the first description of the pulmonary vasculature in both fossil and extant actinistian, a non-tetrapod sarcopterygian clade, contributing to a more in-depth discussion on the morphology of these structures and on the possible homology between vertebrate air-filled organs (lungs of sarcopterygians, lungs of actinopterygians, and gas bladders of actinopterygians).

Life history and ossification patterns in Miguashaia bureaui reveal the early evolution of osteogenesis in coelacanths.

The study of development is critical for revealing the evolution of major vertebrate lineages. Coelacanths have one of the longest evolutionary histories among osteichthyans, but despite access to extant representatives, the onset of their weakly ossified endoskeleton is still poorly understood. Here we present the first palaeohistological and skeletochronological study of Miguashaia bureaui from the Upper Devonian of Canada, pivotal for exploring the palaeobiology and early evolution of osteogenesis in coelacanths. Cross sections of the caudal fin bones show that the cortex is made of layers of primary bone separated by lines of arrested growth, indicative of a cyclical growth. The medullary cavity displays remnants of calcified cartilage associated with bony trabeculae, characteristic of endochondral ossification. A skeletochronological analysis indicates that rapid growth during a short juvenile period was followed by slower growth in adulthood. Our new analysis highlights the life history and palaeoecology of Miguashaia bureaui and reveals that, despite differences in size and habitat, the poor endoskeletal ossification known in the extant Latimeria chalumnae can be traced back at least 375 million years ago.

A microanatomical and histological study of the scales of the Devonian sarcopterygian Miguashaia bureaui and the evolution of the squamation in coelacanths.

Coelacanths have traditionally been described as morphologically conservative throughout their long evolutionary history, which spans more than 400 million years. After an initial burst during the Devonian, a morphological stasis was long thought to have prevailed since the Carboniferous, as shown by the extant Latimeria. New fossil discoveries have challenged this view, with punctual and sometimes unusual departures from the general coelacanth Bauplan. The dermal skeleton is considered to represent one, if not the main, example of morphological stasis in coelacanth evolution and as a consequence, has remained poorly surveyed. The lack of palaeohistological data on the dermoskeleton has resulted in a poor understanding of the early establishment and evolution of the coelacanth squamation. Here we describe the scales of Miguashaia bureaui from the Upper Devonian of Miguasha, Québec (Canada), revealing histological data for a Palaeozoic coelacanth in great detail and adding to our knowledge on the dermal skeleton of sarcopterygians. Miguashaia displays rounded scales ornamented by tubercules and narrow ridges made of dentine and capped with enamel. At least two generations of superimposed odontodes occur, which is reminiscent of the primitive condition of stem osteichthyans like Andreolepis or Lophosteus, and onychodonts like Selenodus. The middle vascular layer is well developed and shows traces of osteonal remodelling. The basal plate consists of a fully mineralised lamellar bone with a repetitive rotation pattern every five layers indicating a twisted plywood-like arrangement of the collagen plies. Comparisons with the extant Latimeria and other extinct taxa show that these features are consistently conserved across coelacanth evolution with only minute changes in certain taxa. The morphological and histological features displayed in the scales of Miguashaia enable us to draw a comprehensive picture of the onset of the coelacanth squamation and to propose and discuss evolutionary scenarios for the coelacanth dermoskeleton.

Diagnosis and management of male urinary tract infections: a need for new guidelines. Study from a French general practice electronic database.

BACKGROUND: The definition and the treatment of male urinary tract infections (UTIs) are imprecise. This study aims to determine the frequency of male UTIs in consultations of general practice, the diagnostic approach and the prescribed treatments. METHODS: We extracted the consultations of male patients, aged 18 years or more, during the period 2012-17 with the International Classification of Primary Care, version 2 codes for UTIs or associated symptoms from PRIMEGE/MEDISEPT databases of primary care. For eligible consultations in which all symptoms or codes were consistent with male UTIs, we identified patient history, prescribed treatments, antibiotic duration, clinical conditions, additional examinations and bacteriological results of urine culture. RESULTS: Our study included 610 consultations with 396 male patients (mean age 62.5 years). Male UTIs accounted for 0.097% of visits and 1.44 visits per physician per year. The UTIs most commonly identified were: undifferentiated (52%), prostatitis (36%), cystitis (8.5%) and pyelonephritis (3.5%). Fever was recorded in 14% of consultations. Urine dipstick test was done in 1.8% of consultations. Urine culture was positive for Escherichia coli in 50.4% of bacteriological tests. Fluoroquinolones were the most prescribed antibiotics (64.9%), followed by beta-lactams (17.4%), trimethoprim-sulfamethoxazole (11.9%) and nitrofurantoin (2.6%). CONCLUSIONS: Male UTIs are rare in general practice and have different presentations. The definition of male UTIs needs to be specified by prospective studies. Diagnostic evidence of male cystitis may reduce the duration of antibiotic therapy and spare critical antibiotics.

The definition and the treatment of male urinary tract infections (UTIs) are imprecise. We aimed to determine the frequency of male UTIs, the diagnostic approach and the prescribed treatments in French electronic health records of general practice. Our study included 610 consultations with 396 male patients with UTIs. In most cases, the organic site of the UTI was not determined. Prostatitis, cystitis and pyelonephritis were diagnosed to a lesser degree. Most patients did not have fever. Half of urine cultures were positive for Escherichia coli, a bacterium from the gastrointestinal tract. Antibiotics were the treatment of choice for male UTIs. In our study, fluoroquinolones (FQs) were the most prescribed antibiotics, then beta-lactams, trimethoprim-sulfamethoxazole and nitrofurantoin. All infections were treated in the same way. Male UTIs are rare in general practice and have different presentations. The resistance of bacteria to FQs is increasing. General practitioners should prescribe antibiotics carefully to avoid failure in the event of recurrent infections. Treating cystitis, prostatitis and pyelonephritis differently may reduce the duration of antibiotic therapy and spare critical antibiotics.

A tomographic study of the histological structure of teeth in the gilthead sea bream, Sparus aurata (Teleostei, Perciformes, Sparidae).

X-ray tomography shows that caniniform and molariform teeth of the gilthead sea bream, Sparus aurata, have a simplexodont plicidentine organization. Together with an insertion of the teeth in alveolae, and the presence of bony shafts sustaining the dental plate, the simplexodont plicidentine is linked to the durophagous diet of the fish.

The phylogenetic origin and evolution of acellular bone in teleost fishes: insights into osteocyte function in bone metabolism.

Vertebrate bone is composed of three main cell types: osteoblasts, osteoclasts and osteocytes, the latter being by far the most numerous. Osteocytes are thought to play a fundamental role in bone physiology and homeostasis, however they are entirely absent in most extant species of teleosts, a group that comprises the vast majority of bony 'fishes', and approximately half of vertebrates. Understanding how this acellular (anosteocytic) bone appeared and was maintained in such an important vertebrate group has important implications for our understanding of the function and evolution of osteocytes. Nevertheless, although it is clear that cellular bone is ancestral for teleosts, it has not been clear in which specific subgroup the osteocytes were lost. This review aims to clarify the phylogenetic distribution of cellular and acellular bone in teleosts, to identify its precise origin, reversals to cellularity, and their implications. We surveyed the bone type for more than 600 fossil and extant ray-finned fish species and optimised the results on recent large-scale molecular phylogenetic trees, estimating ancestral states. We find that acellular bone is a probable synapomorphy of Euteleostei, a group uniting approximately two-thirds of teleost species. We also confirm homoplasy in these traits: acellular bone occurs in some non-euteleosts (although rarely), and cellular bone was reacquired several times independently within euteleosts, in salmons and relatives, tunas and the opah (Lampris sp.). The occurrence of peculiar ecological (e.g. anadromous migration) and physiological (e.g. red-muscle endothermy) strategies in these lineages might explain the reacquisition of osteocytes. Our review supports that the main contribution of osteocytes in teleost bone is to mineral homeostasis (via osteocytic osteolysis) and not to strain detection or bone remodelling, helping to clarify their role in bone physiology.

Histology of the endothermic opah (Lampris sp.) suggests a new structure-function relationship in teleost fish bone.

Endothermy, production and retention of heat by the body, appeared convergently in mammals, birds and four spiny-rayed teleost fish lineages. Of these, red-muscle endothermy over most or all of the body has only appeared in two groups: tunas and the opah (Lampris). Hitherto, tunas have been the only spiny-rayed fishes known to have bones containing embedded osteocyte cells; others have acellular bone. We examined bone histology in Lampris for the first time, demonstrating the presence of cellular bone very similar to that of tunas. This contrasts with the acellular condition of its ectothermic close relatives. The distribution of this character suggests that it co-evolved with red-muscle endothermy, hinting at a common physiological mechanism that would link bone histology to endothermy in these distantly related teleost lineages.

Earliest known lepisosteoid extends the range of anatomically modern gars to the Late Jurassic.

Lepisosteoids are known for their evolutionary conservatism, and their body plan can be traced at least as far back as the Early Cretaceous, by which point two families had diverged: Lepisosteidae, known since the Late Cretaceous and including all living species and various fossils from all continents, except Antarctica and Australia, and Obaichthyidae, restricted to the Cretaceous of northeastern Brazil and Morocco. Until now, the oldest known lepisosteoids were the obaichthyids, which show general neopterygian features lost or transformed in lepisosteids. Here we describe the earliest known lepisosteoid (Nhanulepisosteus mexicanus gen. and sp. nov.) from the Upper Jurassic (Kimmeridgian - about 157 Myr), of the Tlaxiaco Basin, Mexico. The new taxon is based on disarticulated cranial pieces, preserved three-dimensionally, as well as on scales. Nhanulepisosteus is recovered as the sister taxon of the rest of the Lepisosteidae. This extends the chronological range of lepisosteoids by about 46 Myr and of the lepisosteids by about 57 Myr, and fills a major morphological gap in current understanding the early diversification of this group.

The homology and function of the lung plates in extant and fossil coelacanths.

The presence of a pulmonary organ that is entirely covered by true bone tissue and fills most of the abdominal cavity is hitherto unique to fossil actinistians. Although small hard plates have been recently reported in the lung of the extant coelacanth Latimeria chalumnae, the homology between these hard structures in fossil and extant forms remained to be demonstrated. Here, we resolve this question by reporting the presence of a similar histological pattern-true cellular bone with star-shaped osteocytes, and a globular mineralisation with radiating arrangement-in the lung plates of two fossil coelacanths (Swenzia latimerae and Axelrodichthys araripensis) and the plates that surround the lung of the most extensively studied extant coelacanth species, L. chalumnae. The point-for-point structural similarity of the plates in extant and fossil coelacanths supports their probable homology and, consequently, that of the organ they surround. Thus, this evidence questions the previous interpretations of the fatty organ as a component of the pulmonary complex of Latimeria.

†Sorbinicharax verraesi: An unexpected case of a benthic fish outside Acanthomorpha in the Upper Cretaceous of the Tethyan Sea.

†Sorbinicharax verraesi is a marine teleostean fish from the Upper Cretaceous of Nardò (Italy). It was first attributed to the otophysan order Characiformes, which represents potential evidence for the controversial marine origin of the clade. Through a review of all the available material, we demonstrate that this species is not an otophysan since it lacks key structures that would allow for its inclusion in this group. †Sorbinicharax has a body shape that recalls ground fishes classically assigned to Acanthomorpha. However, no unambiguous feature allows us to relate it to this clade. In fact, the presence of cellular bony tissue supports its exclusion from Eurypterygii. Since no feature permits the definitive attribution of †Sorbinicharax to any teleost group, it remains as Teleostei incertae sedis. We infer that the morphology of †Sorbinicharax indicates a benthic ecology. It displays: an anteriorly wide body with enlarged ribs; large pectoral fins, while anal and dorsal fins are reduced; a large head measuring » of the total body length; and a mouth opening dorsally in a high position. Such morphology was so far undescribed in Nardo. It is surprisingly displayed by a non-eurypterygian teleost fish which means by a fish which does not belong to the clades that diversify since the upper Cretaceous and include the extant families that show ground ecomorphologies.

Lung anatomy and histology of the extant coelacanth shed light on the loss of air-breathing during deep-water adaptation in actinistians.

Lungs are specialized organs originated from the posterior pharyngeal cavity and considered as plesiomorphic for osteichthyans, as they are found in extant basal actinopterygians (i.e. Polypterus) and in all major groups of extant sarcopterygians. The presence of a vestigial lung in adult stages of the extant coelacanth Latimeria chalumnae is the result of allometric growth during ontogeny, in relation with long-time adaptation to deep water. Here, we present the first detailed histological and anatomical description of the lung of Latimeria chalumnae, providing new insights into its arrested differentiation in an air-breathing complex, mainly represented by the absence of pneumocytes and of compartmentalization in the latest ontogenetic stages.

Allometric growth in the extant coelacanth lung during ontogenetic development.

Coelacanths are lobe-finned fishes known from the Devonian to Recent that were long considered extinct, until the discovery of two living species in deep marine waters of the Mozambique Channel and Sulawesi. Despite extensive studies, the pulmonary system of extant coelacanths has not been fully investigated. Here we confirm the presence of a lung and discuss its allometric growth in Latimeria chalumnae, based on a unique ontogenetic series. Our results demonstrate the presence of a potentially functional, well-developed lung in the earliest known coelacanth embryo, and its arrested growth at later ontogenetic stages, when the lung is clearly vestigial. The parallel development of a fatty organ for buoyancy control suggests a unique adaptation to deep-water environments. Furthermore, we provide the first evidence for the presence of small, hard, flexible plates around the lung in L. chalumnae, and consider them homologous to the plates of the 'calcified lung' of fossil coelacanths.

Wall-selective probing of double-walled carbon nanotubes using covalent functionalization.

Double-walled carbon nanotubes (DWNTs) present an original coaxial geometry in which the inner wall is naturally protected from the environment by the outer wall. Covalent functionalization is introduced here as an effective approach to investigate DWNT devices. Performed using an aryldiazonium salt, the functionalization is reversible upon thermal annealing and occurs strictly at the surface of the outer wall, leaving the inner wall essentially unaltered by the chemical bonding. Measurements on functionalized DWNT transistors show that the electrical current is carried by the inner wall and provide unambiguous identification of the metallic or semiconducting character of both walls. New insights about current saturation at high bias in DWNTs are also presented as an illustration of new experiments unlocked by the method. The wall-selectivity of the functionalization not only enables selective optical and electrical probing of the DWNTs, but it also paves the way to designing novel electronic devices in which the inner wall is used for electrical transport while the outer wall chemically interacts with the environment.

High performance resonance Raman spectroscopy using volume Bragg gratings as tunable light filters.

We designed a near infrared tunable resonance Raman spectroscopy system based on a tandem of thick volume Bragg gratings (VBGs). VBGs are here the constituents of two light filtering units: a tunable laser line filter (LLF) and a tunable notch filter (NF). When adapted in a micro-Raman setup with a single stage monochromator (1800 gr/mm grating), the tandem of LLF and NF allowed measurements of Raman signals down to +/-20 cm(-1). The good performance and fast tunability of the VBG Raman system was demonstrated on a sulfur powder and on a bulk single-walled carbon nanotube sample through a series of 22 Stokes and anti-Stokes spectra recorded at excitation wavelengths between 800 and 990 nm. The main drawbacks of the setup are the limited spectral range to the near infrared and the small angular acceptance of the filters (approximately 1 mrad), which causes mainly attenuation problems with the NF. The impact of the main limitations is discussed and solutions are provided.

Detection of choline transporter-like 1 protein CTL1 in neuroblastoma x glioma cells and in the CNS, and its role in choline uptake.

Choline is an essential nutrient necessary for synthesis of membrane phospholipids, cell signalling molecules and acetylcholine. The aim of this study was to detect and characterize the choline transporter-like 1 (CTL1/SLC44A1) protein in CNS tissues and the hybrid neuroblastoma x glioma cell line NG108-15, which synthesizes acetylcholine and has high affinity choline transport but does not express the cholinergic high affinity choline transporter 1. The presence of CTL1 protein in NG108-15 cells was confirmed using our antibody G103 which recognizes the C-terminal domain of human CTL1. Three different cognate small interfering RNAs were used to decrease CTL1 mRNA in NG108-15 cells, causing lowered CTL1 protein expression, choline uptake and cell growth. None of the small interfering RNAs influenced carnitine transport, demonstrating the absence of major non-specific effects. In parental C6 cells knockdown of CTL1 also reduced high affinity choline transport. Our results support the concept that CTL1 protein is necessary for the high affinity choline transport which supplies choline for cell growth. The presence of CTL1 protein in rat and human CNS regions, where it is found in neuronal, glial and endothelial cells, suggests that malfunction of this transporter could have important implications in nervous system development and repair following injury, and in neurodegenerative diseases.

Cortisol mobilizes mineral stores from vertebral skeleton in the European eel: an ancestral origin for glucocorticoid-induced osteoporosis?

Endogenous excess cortisol and glucocorticoid (GC) therapy are a major cause of secondary osteoporosis in humans. Intense bone resorption can also be observed in other vertebrates such as migratory teleost fish at the time of reproductive migration and during fasting when large amounts of calcium and phosphate are required. Using a primitive teleost, the European eel, as a model, we investigated whether cortisol could play an ancestral role in the induction of vertebral skeleton demineralization. Different histological and histomorphometric methods were performed on vertebral samples of control and cortisol-treated eels. We demonstrated that cortisol induced a significant bone demineralization of eel vertebrae, as shown by significant decreases of the mineral ratio measured by incineration, and the degree of mineralization measured by quantitative microradiography of vertebral sections. Histology and image analysis of ultrathin microradiographs showed the induction by cortisol of different mechanisms of bone resorption, including periosteocytic osteolysis and osteoclastic resorption. Specificity of cortisol action was investigated by comparison with the effects of sex steroids. Whereas, testosterone had no effect, estradiol induced vertebral skeleton demineralization, an effect related to the stimulated synthesis of vitellogenin (Vg), an oviparous specific phospho-calcio-lipoprotein. By contrast, the cortisol demineralization effect was not related to any stimulation of Vg. This study demonstrates GC-induced bone demineralization in an adult non-mammalian vertebrate, which undergoes natural bone resorption during its life cycle. Our data suggest that the stimulatory action of cortisol on bone loss may represent an ancestral and conserved endocrine regulation in vertebrates.

Thyroid hormone-induced demineralisation of the vertebral skeleton of the eel, Anguilla anguilla.

The role of thyroid hormones (TH) in bone remodelling is controversial. Indeed, in humans, while they are necessary for normal growth and development, their overproduction can induce important mineral bone loss and osteoporosis. Intense bone resorption is a natural phenomenon also observed in some teleosts, during reproductive migration and fasting. Our work aimed at investigating the effects of chronic treatments with TH (thyroxin, T4 or triiodothyronine, T3) on bone resorption in a migratory fish, the European eel (Anguilla anguilla), a representative species of an ancient group of teleosts (Elopomorphs). The incineration method showed that TH induced a significant mineral loss in eel vertebral skeleton. Histology and histophysical (qualitative and quantitative microradiographs) methods were then applied to vertebral sections to determine which types of resorption were induced by TH. Quantitative image analysis of microradiographs showed that TH significantly increased the porosity of the vertebrae, demonstrating the induction of a severe bone loss. Histology revealed the appearance of large osteoclastic lacunae, indicating a stimulation of osteoclastic resorption. Quantitative image analysis of ultrathin microradiographs showed a significant increase of the size of osteocytic lacunae, indicating a stimulation of periosteocytic osteolysis. Finally, quantitative microradiographs indicated a significant fall of mineralisation degree. TH treatments did not stimulate the production of the calcium-bonded lipo-phospho-protein vitellogenin, indicating that TH-induced bone demineralisation was not mediated by any indirect effect on vitellogenesis. Our study demonstrates that TH may participate in the mobilisation of bone mineral stores in the eel, by inducing different types of vertebral bone resorption, such as osteoclastic resorption and periosteocytic osteolysis. These data suggest that the stimulatory action of TH on bone resorption may be an ancient regulatory mechanism in vertebrates.

Synapsin associates with cyclophilin B in an ATP- and cyclosporin A-dependent manner.

Immunophilins are ubiquitous enzymes responsible for proline isomerisation during protein synthesis and for the chaperoning of several membrane proteins. These activities can be blocked by the immunosuppressants cyclosporin A, FK506 and rapamycin. It has been shown that all three immunosuppressants have neurotrophic activity and can modulate neurotransmitter release, but the molecular basis of these effects is currently unknown. Here, we show that synapsin I, a synaptic vesicle-associated protein, can be purified from Torpedo cholinergic synaptosomes through its affinity to cyclophilin B, an immunophilin that is particularly abundant in brain. The interaction is direct and conserved in mammals, and shows a dissociation constant of about 0.5 microM in vitro. The binding between the two proteins can be disrupted by cyclosporin A and inhibited by physiological concentrations of ATP. Furthermore, cyclophilin B co-localizes with synapsin I in rat synaptic vesicle fractions and its levels in synaptic vesicle-containing fractions are decreased in synapsin knockout mice. These results suggest that immunophilins are involved in the complex protein networks operating at the presynaptic level and implicate the interaction between cyclophilin B and synapsins in presynaptic function.

Structural peculiarities of the tubercles in the skin of the turbot, Scophthalmus maximus (L., 1758) (Osteichthyes, Pleuronectiformes, Scophthalmidae).

The structure of the bony tubercles of the turbot, Scophthalmus maximus (L., 1758), was examined using ground sections, microradiography, SEM, and TEM. The tubercles are small, isolated, mineralized conical plates randomly distributed in the eyed side of the body. They are composed of three layers: the outer limiting layer, the external layer, and the basal plate, which make up the thin and flat elasmoid scales of Teleostei. The main difference between regular elasmoid scales and bony tubercles lies in the organization and the growth of the basal plate. Indeed, the conical shape of the tubercle is the result of a prominent thickening of the central part of the basal plate where the collagen matrix is organized in a complicated three-dimensional network. Densely packed thick collagen fibrils form superimposed plies organized in a plywood-like structure that resembles that of the elasmoid scales but it is criss-crossed by numerous vertical sheets of thin collagen fibrils. The tubercles originate from thin and flat plates located in the skin of larvae and juveniles, whose structure is that of regular-developing elasmoid scales. Thus, the tubercles of Scophthalmus maximus could be considered as modified elasmoid scales rather than bony structures. They might be the result of specific arrangements related to the general trend of reduction of the dermal skeleton in the teleostean lineage.

Selection and characterization of the choline transport mutation suppressor from Torpedo electric lobe, CTL1.

The presumptive choline transporter, CTL1, was initially identified through functional complementation of a triple yeast mutant (ctr ise URA3delta) with deficiencies in both choline transport and choline neosynthesis under selective conditions that cause perturbations in membrane synthesis and growth. After transformation of these yeasts with a heterologous yeast expression library made from Torpedo electric lobe cDNAs, several colonies showed increased growth but only one clone increased the accumulation of external choline. The corresponding full-length cDNA was isolated and encodes a protein with 10 transmembrane domains. Northern analysis of Torpedo mRNA indicates that CTL1 is expressed at high levels in the spinal cord and brain. In Xenopus oocytes, Torpedo CTL1 expression was associated with the appearance of sodium independent high-affinity choline uptake. We propose that CTL1 plays a role in providing choline for membrane synthesis in the nervous system.