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Sample records for intact mouse vomeronasal

  1. Responses to sulfated steroids of female mouse vomeronasal sensory neurons

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    Celsi, F.; d'Errico, A.; Menini, A.

    2012-01-01

    The rodent vomeronasal organ plays an important role in many social behaviors. Using the calcium imaging technique with the dye fluo-4 we measured intracellular calcium concentration changes induced by the application of sulfated steroids to neurons isolated from the vomeronasal organ of female mice. We found that a mix of 10 sulfated steroids from the androgen, estrogen, pregnanolone, and glucocorticoid families induced a calcium response in 71% of neurons. Moreover, 31% of the neurons respo...

  2. Signaling mechanisms and behavioral function of the mouse basal vomeronasal neuroepithelium

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    Anabel ePérez-Gómez

    2014-11-01

    Full Text Available The vomeronasal organ (VNO is a sensory organ that is found in most terrestrial vertebrates and that is principally implicated in the detection of pheromones. The VNO contains specialized sensory neurons organized in a pseudostratified neuroepithelium that recognize chemical signals involved in initiating innate behavioral responses. In rodents, the VNO neuroepithelium is segregated into two distinct zones, apical and basal. The molecular mechanisms involved in ligand detection by apical and basal VNO sensory neurons differ extensively. These two VNO subsystems express different subfamilies of vomeronasal receptors and signaling molecules, detect distinct chemosignals, and project to separate regions of the accessory olfactory bulb (AOB. The roles that these olfactory subdivisions play in the control of specific olfactory-mediated behaviors are largely unclear. However, analysis of mutant mouse lines for signal transduction components together with identification of defined chemosensory ligands has revealed a fundamental role of the basal part of the mouse VNO in mediating a wide range of instinctive behaviors, such as aggression, predator avoidance, and sexual attraction. Here we will compare the divergent functions and synergies between the olfactory subsystems and consider new insights in how higher neural circuits are defined for the initiation of instinctive behaviors.

  3. Sex- and strain-specific expression and vomeronasal activity of mouse ESP family peptides.

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    Kimoto, Hiroko; Sato, Koji; Nodari, Francesco; Haga, Sachiko; Holy, Timothy E; Touhara, Kazushige

    2007-11-06

    Male mice secrete exocrine-gland-secreting peptide 1 (ESP1) from the extraorbital lacrimal gland into tear fluid [1]. Other mice detect ESP1 through sensory neurons in the vomeronasal organ (VNO), a secondary olfactory system that senses pheromonal information, including sex, strain, and species. ESP1 is now known to be a member of a multigene family that encodes peptides of various lengths. We herein performed genomic and expression analyses of the ESP family. The ESP family consists of 38 members in mice and 10 members in rat but is absent from the human genome, suggesting rapid molecular evolution. In addition to the male-specific ESP1, we discovered one, which we designated ESP36, that, in adult BALB/c mice, is expressed only in the female extraorbital lacrimal gland. The sexually dimorphic expression is ensured by the release of testosterone after puberty. However, we observed dramatic differences in the expression levels of ESPs between strains. Finally, all ESPs elicited an electrical response in the vomeronasal epithelium but not in the main olfactory epithelium. Multielectrode recording of VNO activity demonstrated that ESP1 induces action potentials in vomeronasal neurons, leading to an increase in the spike firing rate, and that ESP1 is recognized by narrowly tuned vomeronasal sensory neurons. Sexual dimorphism and strain differences of ESPs and their reception in the VNO suggest that the ESP family can convey information about sex and individual identity via the vomeronasal system. The chemosensation of this nonvolatile peptide family by direct contact appears to be one of strategies for sociosexual communication in rodent species.

  4. NanoCAGE analysis of the mouse olfactory epithelium identifies the expression of vomeronasal receptors and of proximal LINE elements

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    Giovanni ePascarella

    2014-02-01

    Full Text Available By coupling laser capture microdissection to nanoCAGE technology and next-generation sequencing we have identified the genome-wide collection of active promoters in the mouse Main Olfactory Epithelium (MOE. Transcription start sites (TSSs for the large majority of Olfactory Receptors (ORs have been previously mapped increasing our understanding of their promoter architecture.Here we show that in our nanoCAGE libraries of the mouse MOE we detect a large number of tags mapped in loci hosting Type-1 and Type-2 Vomeronasal Receptors genes (V1Rs and V2Rs. These loci also show a massive expression of Long Interspersed Nuclear Elements (LINEs. We have validated the expression of selected receptors detected by nanoCAGE with in situ hybridization, RT-PCR and qRT-PCR. This work extends the repertory of receptors capable of sensing chemical signals in the MOE, suggesting intriguing interplays between MOE and VNO for pheromone processing. It positions transcribed LINEs as candidate regulatory RNAs for VRs expression.

  5. Chemoreception regulates chemical access to mouse vomeronasal organ: role of solitary chemosensory cells.

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    Tatsuya Ogura

    Full Text Available Controlling stimulus access to sensory organs allows animals to optimize sensory reception and prevent damage. The vomeronasal organ (VNO detects pheromones and other semiochemicals to regulate innate social and sexual behaviors. This semiochemical detection generally requires the VNO to draw in chemical fluids, such as bodily secretions, which are complex in composition and can be contaminated. Little is known about whether and how chemical constituents are monitored to regulate the fluid access to the VNO. Using transgenic mice and immunolabeling, we found that solitary chemosensory cells (SCCs reside densely at the entrance duct of the VNO. In this region, most of the intraepithelial trigeminal fibers innervate the SCCs, indicating that SCCs relay sensory information onto the trigeminal fibers. These SCCs express transient receptor potential channel M5 (TRPM5 and the phospholipase C (PLC beta2 signaling pathway. Additionally, the SCCs express choline acetyltransferase (ChAT and vesicular acetylcholine transporter (VAChT for synthesizing and packaging acetylcholine, a potential transmitter. In intracellular Ca2+ imaging, the SCCs responded to various chemical stimuli including high concentrations of odorants and bitter compounds. The responses were suppressed significantly by a PLC inhibitor, suggesting involvement of the PLC pathway. Further, we developed a quantitative dye assay to show that the amount of stimulus fluid that entered the VNOs of behaving mice is inversely correlated to the concentration of odorous and bitter substances in the fluid. Genetic knockout and pharmacological inhibition of TRPM5 resulted in larger amounts of bitter compounds entering the VNOs. Our data uncovered that chemoreception of fluid constituents regulates chemical access to the VNO and plays an important role in limiting the access of non-specific irritating and harmful substances. Our results also provide new insight into the emerging role of SCCs in

  6. Genomic organization and sequence analysis of the vomeronasal receptor V2R genes in mouse genome

    Institute of Scientific and Technical Information of China (English)

    YANG Hui; Zhang YaPing

    2007-01-01

    Two multigene superfamilies, named V1R and V2R, encoding seven-transmembrane-domain G-protein coupled receptors (GPCRs) have been identified as pheromone receptors in mammals. Three V2R gene families have been described in mouse and rat. Here we screened the updated mouse genome sequence database and finally retrieved 63 putative functional V2R genes including three newly identified genes which formed a new additional family. We described the genomic organization of these genes and also characterized the conservation of mouse V2R protein sequences. These genomic and sequence information we described are useful as part of the evidence to speculate the functional domain of V2Rs and should give aid to the functionality study in the future.

  7. Tuning Properties and Dynamic Range of Type 1 Vomeronasal Receptors

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    Sachiko eHaga-Yamanaka

    2015-07-01

    Full Text Available The mouse vomeronasal organ expresses chemosensory receptors that detect intra-species as well as inter-species cues. The vomeronasal neurons are thought to be highly selective in their responses. The tuning properties of individual receptors remain difficult to characterize due to the lack of a robust heterologous expression system. Here, we take a transgenic approach to ectopically express two Type 1 vomeronasal receptors in the mouse vomeronasal organ and characterize their responses to steroid compounds. We find that V1rj2 and V1rj3 are sensitive to two sulfated estrogens and can be activated by a broad variety of sulfated and glucuronidated steroids at high concentrations. Individual neurons exhibit narrow range of concentration-dependent activation. Collectively, a neuronal population expressing the same receptor covers a wide dynamic range in their responses to sulfated estrogens. These properties recapitulate the response profiles of endogenous neurons to sulfated estrogens.

  8. Mechanisms of neuronal chloride accumulation in intact mouse olfactory epithelium.

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    Nickell, William T; Kleene, Nancy K; Kleene, Steven J

    2007-09-15

    When olfactory receptor neurons respond to odours, a depolarizing Cl(-) efflux is a substantial part of the response. This requires that the resting neuron accumulate Cl(-) against an electrochemical gradient. In isolated olfactory receptor neurons, the Na(+)-K(+)-2Cl(-) cotransporter NKCC1 is essential for Cl(-) accumulation. However, in intact epithelium, a robust electrical olfactory response persists in mice lacking NKCC1. This response is largely due to a neuronal Cl(-) efflux. It thus appears that NKCC1 is an important part of a more complex system of Cl(-) accumulation. To identify the remaining transport proteins, we first screened by RT-PCR for 21 Cl(-) transporters in mouse nasal tissue containing olfactory mucosa. For most of the Cl(-) transporters, the presence of mRNA was demonstrated. We also investigated the effects of pharmacological block or genetic ablation of Cl(-) transporters on the olfactory field potential, the electroolfactogram (EOG). Mice lacking the common Cl(-)/HCO(3)(-) exchanger AE2 had normal EOGs. Block of NKCC cotransport with bumetanide reduced the EOG in epithelia from wild-type mice but had no effect in mice lacking NKCC1. Hydrochlorothiazide, a blocker of the Na(+)-Cl(-) cotransporter, had only a small effect. DIDS, a blocker of some KCC cotransporters and Cl(-)/HCO(3)(-) exchangers, reduced the EOG in epithelia from both wild-type and NKCC1 knockout mice. A combination of bumetanide and DIDS decreased the response more than either drug alone. However, no combination of drugs completely abolished the Cl(-) component of the response. These results support the involvement of both NKCC1 and one or more DIDS-sensitive transporters in Cl(-) accumulation in olfactory receptor neurons.

  9. Interleukin 6 in intact and injured mouse peripheral nerves.

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    Reichert, F; Levitzky, R; Rotshenker, S

    1996-03-01

    The multifunctional cytokine interleukin 6 (IL-6) has direct growth, survival and differentiation effects on peripheral and central neurons. Furthermore, it can modulate the production by non-neuronal cells of other cytokines and growth factors, and thereby affect nerve cells indirectly. We have studied IL-6 expression and production in intact and injured peripheral nerves of C57/BL/6NHSD mice, which display the normal rapid progression of Wallerian degeneration. The IL-6 mRNA was detected in nerves degenerating in vitro or in vivo, but not in intact nerves. In vitro- and in vivo-degenerating nerve segments and neuroma nerve segments synthesized and secreted IL-6. The onset of IL-6 production was rapid and prolonged. It was detected as early as 2 h after injury and persisted for the entire period of 21 days tested after the injury. Of the non-neuronal cells that reside in intact and injured nerves, macrophages and fibroblasts were the major contributors to IL-6 production. We also studied IL-6 production in intact and injured nerves of mutant C57BL/6-WLD/OLA/NHSD mice, which display very slow progression of Wallerian degeneration. Injured nerves of C57BL/6-WLD/OLA/NHSD mice produced significantly lower amounts of IL-6 than did rapidly degenerating nerves of C57/BL/6NHSD mice.

  10. Tuning properties and dynamic range of type 1 vomeronasal receptors

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    Haga-Yamanaka, Sachiko; Ma, Limei; Yu, C. Ron

    2015-01-01

    The mouse vomeronasal organ (VNO) expresses chemosensory receptors that detect intra-species as well as inter-species cues. The vomeronasal neurons are thought to be highly selective in their responses. The tuning properties of individual receptors remain difficult to characterize due to the lack of a robust heterologous expression system. Here, we take a transgenic approach to ectopically express two type 1 vomeronasal receptors in the mouse VNO and characterize their responses to steroid compounds. We find that V1rj2 and V1rj3 are sensitive to two sulfated estrogens (SEs) and can be activated by a broad variety of sulfated and glucuronidated steroids at high concentrations. Individual neurons exhibit narrow range of concentration-dependent activation. Collectively, a neuronal population expressing the same receptor covers a wide dynamic range in their responses to SEs. These properties recapitulate the response profiles of endogenous neurons to SEs. PMID:26236183

  11. Strain-specific Loss of Formyl Peptide Receptor 3 in the Murine Vomeronasal and Immune Systems.

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    Stempel, Hendrik; Jung, Martin; Pérez-Gómez, Anabel; Leinders-Zufall, Trese; Zufall, Frank; Bufe, Bernd

    2016-04-29

    Formyl peptide receptor 3 (Fpr3, also known as Fpr-rs1) is a G protein-coupled receptor expressed in subsets of sensory neurons of the mouse vomeronasal organ, an olfactory substructure essential for social recognition. Fpr3 has been implicated in the sensing of infection-associated olfactory cues, but its expression pattern and function are incompletely understood. To facilitate visualization of Fpr3-expressing cells, we generated and validated two new anti-Fpr3 antibodies enabling us to analyze acute Fpr3 protein expression. Fpr3 is not only expressed in murine vomeronasal sensory neurons but also in bone marrow cells, the primary source for immune cell renewal, and in mature neutrophils. Consistent with the notion that Fpr3 functions as a pathogen sensor, Fpr3 expression in the immune system is up-regulated after stimulation with a bacterial endotoxin (lipopolysaccharide). These results strongly support a dual role for Fpr3 in both vomeronasal sensory neurons and immune cells. We also identify a large panel of mouse strains with severely altered expression and function of Fpr3, thus establishing the existence of natural Fpr3 knock-out strains. We attribute distinct Fpr3 expression in these strains to the presence or absence of a 12-nucleotide in-frame deletion (Fpr3Δ424-435). In vitro calcium imaging and immunofluorescence analyses demonstrate that the lack of four amino acids leads to an unstable, truncated, and non-functional receptor protein. The genome of at least 19 strains encodes a non-functional Fpr3 variant, whereas at least 13 other strains express an intact receptor. These results provide a foundation for understanding the in vivo function of Fpr3. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  12. Label-free structural photoacoustic tomography of intact mouse brain

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    Li, Lei; Xia, Jun; Li, Guo; Garcia-Uribe, Alejandro; Wang, Lihong V.

    2015-03-01

    Capitalizing on endogenous hemoglobin contrast, photoacoustic computed tomography (PACT), a deep-tissue highresolution imaging modality, has drawn increasing interest in neuro-imaging. However, most existing studies are limited to functional imaging on the cortical surface, and the deep-brain structural imaging capability of PACT has never been demonstrated. Here, we explicitly studied the limiting factors of deep-brain PACT imaging. We found that the skull distorted the acoustic signal and blood suppressed the structural contrast from other chromophores. When the two effects are mitigated, PACT can provide high-resolution label-free structural imaging through the entire mouse brain. With 100 μm in-plane resolution, we can clearly identify major structures of the brain, and the image quality is comparable to that of magnetic resonance microscopy. Spectral PACT studies indicate that structural contrasts mainly originate from cytochrome and lipid. The feasibility of imaging the structure of the brain in vivo has also been discussed. Our results demonstrate that PACT is a promising modality for both structural and functional brain imaging.

  13. Conserved repertoire of orthologous vomeronasal type 1 receptor genes in ruminant species

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    Okamura Hiroaki

    2009-09-01

    Full Text Available Abstract Background In mammals, pheromones play an important role in social and innate reproductive behavior within species. In rodents, vomeronasal receptor type 1 (V1R, which is specifically expressed in the vomeronasal organ, is thought to detect pheromones. The V1R gene repertoire differs dramatically between mammalian species, and the presence of species-specific V1R subfamilies in mouse and rat suggests that V1R plays a profound role in species-specific recognition of pheromones. In ruminants, however, the molecular mechanism(s for pheromone perception is not well understood. Interestingly, goat male pheromone, which can induce out-of-season ovulation in anestrous females, causes the same pheromone response in sheep, and vice versa, suggesting that there may be mechanisms for detecting "inter-species" pheromones among ruminant species. Results We isolated 23 goat and 21 sheep intact V1R genes based on sequence similarity with 32 cow V1R genes in the cow genome database. We found that all of the goat and sheep V1R genes have orthologs in their cross-species counterparts among these three ruminant species and that the sequence identity of V1R orthologous pairs among these ruminants is much higher than that of mouse-rat V1R orthologous pairs. Furthermore, all goat V1Rs examined thus far are expressed not only in the vomeronasal organ but also in the main olfactory epithelium. Conclusion Our results suggest that, compared with rodents, the repertoire of orthologous V1R genes is remarkably conserved among the ruminants cow, sheep and goat. We predict that these orthologous V1Rs can detect the same or closely related chemical compound(s within each orthologous set/pair. Furthermore, all identified goat V1Rs are expressed in the vomeronasal organ and the main olfactory epithelium, suggesting that V1R-mediated ligand information can be detected and processed by both the main and accessory olfactory systems. The fact that ruminant and rodent V1Rs

  14. Formyl Peptide Receptors from Immune and Vomeronasal System Exhibit Distinct Agonist Properties*

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    Bufe, Bernd; Schumann, Timo; Zufall, Frank

    2012-01-01

    The formyl peptide receptor (Fpr) family is well known for its contribution to immune defense against pathogens in human and rodent leukocytes. Recently, several structurally related members of these receptors were discovered in sensory neurons of the mouse vomeronasal organ (VNO), key detectors of pheromones and related semiochemicals. Although the biological role of vomeronasal Fprs is not yet clear, the known contribution of other Fprs to host immune defense suggested that they could contribute to vomeronasal pathogen sensing. Precise knowledge about the agonist properties of mouse Fprs is required to determine their function. We expressed all seven mouse and three human Fprs using an in vitro system and tested their activation with 32 selected compounds by conducting high throughput calcium measurements. We found an intriguing functional conservation between human and mouse immune Fprs that is most likely a consequence of closely similar biological constraints. By contrast, our data suggest a neofunctionalization of the vomeronasal Fprs. We show that the vomeronasal receptor mFpr-rs1 can be activated robustly by W-peptide and structural derivatives but not by other typical ligands of immune Fprs. mFpr-rs1 exhibits a stereo-selective preference for peptides containing d-amino acids. The same peptide motifs are contained in pathogenic microorganisms. Thus, the ligand profile of mFpr-rs1 is consistent with a role in vomeronasal pathogen sensing. PMID:22859307

  15. Vomeronasal organ and human pheromones.

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    Trotier, D

    2011-09-01

    For many organisms, pheromonal communication is of particular importance in managing various aspects of reproduction. In tetrapods, the vomeronasal (Jacobson's) organ specializes in detecting pheromones in biological substrates of congeners. This information triggers behavioral changes associated, in the case of certain pheromones, with neuroendocrine correlates. In human embryos, the organ develops and the nerve fibers constitute a substrate for the migration of GnRH-secreting cells from the olfactory placode toward the hypothalamus. After this essential step for subsequent secretion of sex hormones by the anterior hypophysis, the organ regresses and the neural connections disappear. The vomeronasal cavities can still be observed by endoscopy in some adults, but they lack sensory neurons and nerve fibers. The genes which code for vomeronasal receptor proteins and the specific ionic channels involved in the transduction process are mutated and nonfunctional in humans. In addition, no accessory olfactory bulbs, which receive information from the vomeronasal receptor cells, are found. The vomeronasal sensory function is thus nonoperational in humans. Nevertheless, several steroids are considered to be putative human pheromones; some activate the anterior hypothalamus, but the effects observed are not comparable to those in other mammals. The signaling process (by neuronal detection and transmission to the brain or by systemic effect) remains to be clearly elucidated.

  16. Fluorescent-protein stabilization and high-resolution imaging of cleared, intact mouse brains.

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    Martin K Schwarz

    Full Text Available In order to observe and quantify long-range neuronal connections in intact mouse brain by light microscopy, it is first necessary to clear the brain, thus suppressing refractive-index variations. Here we describe a method that clears the brain and preserves the signal from proteinaceous fluorophores using a pH-adjusted non-aqueous index-matching medium. Successful clearing is enabled through the use of either 1-propanol or tert-butanol during dehydration whilst maintaining a basic pH. We show that high-resolution fluorescence imaging of entire, structurally intact juvenile and adult mouse brains is possible at subcellular resolution, even following many months in clearing solution. We also show that axonal long-range projections that are EGFP-labelled by modified Rabies virus can be imaged throughout the brain using a purpose-built light-sheet fluorescence microscope. To demonstrate the viability of the technique, we determined a detailed map of the monosynaptic projections onto a target cell population in the lateral entorhinal cortex. This example demonstrates that our method permits the quantification of whole-brain connectivity patterns at the subcellular level in the uncut brain.

  17. Cladistic Analysis of Olfactory and Vomeronasal Systems

    OpenAIRE

    Ubeda-Bañon, Isabel; Pro-Sistiaga, Palma; Mohedano-Moriano, Alicia; Saiz-Sanchez, Daniel; de la Rosa-Prieto, Carlos; Gutierrez-Castellanos, Nicolás; Lanuza, Enrique; Martinez-Garcia, Fernando; Martinez-Marcos, Alino

    2011-01-01

    Most tetrapods possess two nasal organs for detecting chemicals in their environment, which are the sensory detectors of the olfactory and vomeronasal systems. The seventies’ view that the olfactory system was only devoted to sense volatiles, whereas the vomeronasal system was exclusively specialized for pheromone detection was challenged by accumulating data showing deep anatomical and functional interrelationships between both systems. In addition, the assumption that the vomeronasal system...

  18. Cladistic analysis of olfactory and vomeronasal systems

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    Alino eMartinez-Marcos

    2011-01-01

    Most tetrapods possess two nasal organs for detecting chemicals in their environment, which are the sensory detectors of the olfactory and vomeronasal systems. The seventies’ view that the olfactory system was only devoted to sense volatiles, whereas the vomeronasal system was exclusively specialized for pheromone detection was challenged by accumulating data showing deep anatomical and functional interrelationships between both systems. In addition, the assumption that the vomeronasal system...

  19. An effective manual deboning method to prepare intact mouse nasal tissue with preserved anatomical organization.

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    Dunston, David; Ashby, Sarah; Krosnowski, Kurt; Ogura, Tatsuya; Lin, Weihong

    2013-08-10

    The mammalian nose is a multi-functional organ with intricate internal structures. The nasal cavity is lined with various epithelia such as olfactory, respiratory, and squamous epithelia which differ markedly in anatomical locations, morphology, and functions. In adult mice, the nose is covered with various skull bones, limiting experimental access to internal structures, especially those in the posterior such as the main olfactory epithelium (MOE). Here we describe an effective method for obtaining almost the entire and intact nasal tissues with preserved anatomical organization. Using surgical tools under a dissecting microscope, we sequentially remove the skull bones surrounding the nasal tissue. This procedure can be performed on both paraformaldehyde-fixed and freshly dissected, skinned mouse heads. The entire deboning procedure takes about 20-30 min, which is significantly shorter than the experimental time required for conventional chemical-based decalcification. In addition, we present an easy method to remove air bubbles trapped between turbinates, which is critical for obtaining intact thin horizontal or coronal or sagittal sections from the nasal tissue preparation. Nasal tissue prepared using our method can be used for whole mount observation of the entire epithelia, as well as morphological, immunocytochemical, RNA in situ hybridization, and physiological studies, especially in studies where region-specific examination and comparison are of interest.

  20. Simultaneous high-speed imaging and optogenetic inhibition in the intact mouse brain

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    Bovetti, Serena; Moretti, Claudio; Zucca, Stefano; Dal Maschio, Marco; Bonifazi, Paolo; Fellin, Tommaso

    2017-01-01

    Genetically encoded calcium indicators and optogenetic actuators can report and manipulate the activity of specific neuronal populations. However, applying imaging and optogenetics simultaneously has been difficult to establish in the mammalian brain, even though combining the techniques would provide a powerful approach to reveal the functional organization of neural circuits. Here, we developed a technique based on patterned two-photon illumination to allow fast scanless imaging of GCaMP6 signals in the intact mouse brain at the same time as single-photon optogenetic inhibition with Archaerhodopsin. Using combined imaging and electrophysiological recording, we demonstrate that single and short bursts of action potentials in pyramidal neurons can be detected in the scanless modality at acquisition frequencies up to 1 kHz. Moreover, we demonstrate that our system strongly reduces the artifacts in the fluorescence detection that are induced by single-photon optogenetic illumination. Finally, we validated our technique investigating the role of parvalbumin-positive (PV) interneurons in the control of spontaneous cortical dynamics. Monitoring the activity of cellular populations on a precise spatiotemporal scale while manipulating neuronal activity with optogenetics provides a powerful tool to causally elucidate the cellular mechanisms underlying circuit function in the intact mammalian brain. PMID:28053310

  1. Simultaneous high-speed imaging and optogenetic inhibition in the intact mouse brain

    Science.gov (United States)

    Bovetti, Serena; Moretti, Claudio; Zucca, Stefano; Dal Maschio, Marco; Bonifazi, Paolo; Fellin, Tommaso

    2017-01-01

    Genetically encoded calcium indicators and optogenetic actuators can report and manipulate the activity of specific neuronal populations. However, applying imaging and optogenetics simultaneously has been difficult to establish in the mammalian brain, even though combining the techniques would provide a powerful approach to reveal the functional organization of neural circuits. Here, we developed a technique based on patterned two-photon illumination to allow fast scanless imaging of GCaMP6 signals in the intact mouse brain at the same time as single-photon optogenetic inhibition with Archaerhodopsin. Using combined imaging and electrophysiological recording, we demonstrate that single and short bursts of action potentials in pyramidal neurons can be detected in the scanless modality at acquisition frequencies up to 1 kHz. Moreover, we demonstrate that our system strongly reduces the artifacts in the fluorescence detection that are induced by single-photon optogenetic illumination. Finally, we validated our technique investigating the role of parvalbumin-positive (PV) interneurons in the control of spontaneous cortical dynamics. Monitoring the activity of cellular populations on a precise spatiotemporal scale while manipulating neuronal activity with optogenetics provides a powerful tool to causally elucidate the cellular mechanisms underlying circuit function in the intact mammalian brain.

  2. 3D perfusion mapping in the intact mouse heart after myocardial infarction using myocardial contrast echocardiography

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    Li, Yinbo; Yang, Zequan; French, Brent A.; Hossack, John A.

    2005-04-01

    An intact mouse model of surgically-induced myocardial infarction (MI) caused by permanent occlusion of the Left Anterior Descending (LAD) coronary artery was studied. Normal mice with no occlusion were also studied as controls. For each mouse, contrast enhanced ultrasound images of the heart were acquired in parallel cross-sections perpendicular to the sternum at millimeter increments. For accurate 3D reconstruction, ECG gating and a tri-axial adjustable micromanipulator were used for temporal and spatial registration. Ultrasound images at steady-state of blood refilling were color-coded in each slice to show relative perfusion. Myocardial perfusion defects and necrosis were also examined postmortem by staining with Phthalo blue and TTC red dyes. Good correlation (R>0.93) in perfused area size was observed between in vivo measurements and histological staining. A 3D multi-slice model and a 3D rendering of perfusion distribution were created and showed a promising match with postmortem results, lending further credence to its use as a more comprehensive and more reliable tool for in vivo assessment of myocardial perfusion than 2D tomographic analysis.

  3. Imaging of the intact mouse cochlea by spectral domain optical coherence tomography

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    Gao, Simon S.; Yuan, Tao; Xia, Anping; Raphael, Patrick; Shelton, Ryan L.; Applegate, Brian E.; Oghalai, John S.

    2011-03-01

    Current medical imaging modalities, such as MRI and CT, do not provide high enough resolution to detect many changes within the cochlea that cause hearing loss. We sought to develop the technique of optical coherence tomography (OCT) to image the cochlea noninvasively and within its native environment. We used spectral domain OCT with 950 nm as the center wavelength and a bandwidth of ~100 nm to image freshly excised normal mouse cochlea at different developmental ages. The OCT system has an axial resolution of ~4 μm (in air) and a lateral resolution of ~10 μm. When we imaged normal adult mouse cochleae through the round window membrane, Reissner's membrane, the basilar membrane, the tectorial membrane, the spiral ligament, the spiral limbus, and the modiolus could be clearly identified. When we imaged intact adult cochleae, we were able to image through ~130 μm of bone and tissue to see up to a depth of ~600 μm, and all of the previously identified structures were still visible. Imaging of early postnatal mice during the timeline of cochlear development permitted visualization of the expected structural differences from adult cochleae. Therefore, we conclude that spectral domain OCT is an effective technique for noninvasive imaging of the murine cochlea.

  4. Intact fetal ovarian cord formation promotes mouse oocyte survival and development

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    Pera Renee

    2010-01-01

    Full Text Available Abstract Background Female reproductive potential, or the ability to propagate life, is limited in mammals with the majority of oocytes lost before birth. In mice, surviving perinatal oocytes are enclosed in ovarian follicles for subsequent oocyte development and function in the adult. Before birth, fetal germ cells of both sexes develop in clusters, or germline cysts, in the undifferentiated gonad. Upon sex determination of the fetal gonad, germ cell cysts become organized into testicular or ovarian cord-like structures and begin to interact with gonadal somatic cells. Although germline cysts and testicular cords are required for spermatogenesis, the role of cyst and ovarian cord formation in mammalian oocyte development and female fertility has not been determined. Results Here, we examine whether intact fetal ovarian germ and somatic cell cord structures are required for oocyte development using mouse gonad re-aggregation and transplantation to disrupt gonadal organization. We observed that germ cells from disrupted female gonad prior to embryonic day e13.5 completed prophase I of meiosis but did not survive following transplantation. Furthermore, re-aggregated ovaries from e13.5 to e15.5 developed with a reduced number of oocytes. Oocyte loss occurred before follicle formation and was associated with an absence of ovarian cord structure and ovary disorganization. However, disrupted ovaries from e16.5 or later were resistant to the re-aggregation impairment and supported robust oocyte survival and development in follicles. Conclusions Thus, we demonstrate a critical window of oocyte development from e13.5 to e16.5 in the intact fetal mouse ovary, corresponding to the establishment of ovarian cord structure, which promotes oocyte interaction with neighboring ovarian somatic granulosa cells before birth and imparts oocytes with competence to survive and develop in follicles. Because germline cyst and ovarian cord structures are conserved in the

  5. Role of the vomeronasal organ on the estral cycle reduction by pheromones in the rat.

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    Mora, O A; Sánchez-Criado, J E; Guisado, S

    1985-09-01

    The role of he vomeronasal organ on the estral cycle reduction induced by pheromones is studied in adult female wistar rats. The animals were divided in three groups: I, intact rats; II, vomeronasalectomized rats (VNX); and III, sham operated rats (sham). Each group was submitted to another three distinct conditions from the day they were weaned (21 days old): Isolated female rats; with male odors from two adult males of tested sexual potency, and isolated rats again. The isolated intact rats show mainly 5 day length cycles. The groups I and III (intacts and sham) with male odors, show 4 day length cycles. The VNX animals show 5 day cycles in any one experimental conditions. These results support the idea that the vomeronasal organ is the receptor of the male reducing cycle pheromone in the female rat.

  6. Effect of temperature on crossbridge force changes during fatigue and recovery in intact mouse muscle fibers.

    Directory of Open Access Journals (Sweden)

    Marta Nocella

    Full Text Available Repetitive or prolonged muscle contractions induce muscular fatigue, defined as the inability of the muscle to maintain the initial tension or power output. In the present experiments, made on intact fiber bundles from FDB mouse, fatigue and recovery from fatigue were investigated at 24°C and 35°C. Force and stiffness were measured during tetani elicited every 90 s during the pre-fatigue control phase and recovery and every 1.5 s during the fatiguing phase made of 105 consecutive tetani. The results showed that force decline could be split in an initial phase followed by a later one. Loss of force during the first phase was smaller and slower at 35°C than at 24°C, whereas force decline during the later phase was greater at 35°C so that total force depression at the end of fatigue was the same at both temperatures. The initial force decline occurred without great reduction of fiber stiffness and was attributed to a decrease of the average force per attached crossbridge. Force decline during the later phase was accompanied by a proportional stiffness decrease and was attributed to a decrease of the number of attached crossbridge. Similarly to fatigue, at both 24 and 35°C, force recovery occurred in two phases: the first associated with the recovery of the average force per attached crossbridge and the second due to the recovery of the pre-fatigue attached crossbridge number. These changes, symmetrical to those occurring during fatigue, are consistent with the idea that, i initial phase is due to the direct fast inhibitory effect of [Pi]i increase during fatigue on crossbridge force; ii the second phase is due to the delayed reduction of Ca(2+ release and /or reduction of the Ca(2+ sensitivity of the myofibrils due to high [Pi]i.

  7. Cladistic Analysis of Olfactory and Vomeronasal Systems

    Science.gov (United States)

    Ubeda-Bañon, Isabel; Pro-Sistiaga, Palma; Mohedano-Moriano, Alicia; Saiz-Sanchez, Daniel; de la Rosa-Prieto, Carlos; Gutierrez-Castellanos, Nicolás; Lanuza, Enrique; Martinez-Garcia, Fernando; Martinez-Marcos, Alino

    2010-01-01

    Most tetrapods possess two nasal organs for detecting chemicals in their environment, which are the sensory detectors of the olfactory and vomeronasal systems. The seventies’ view that the olfactory system was only devoted to sense volatiles, whereas the vomeronasal system was exclusively specialized for pheromone detection was challenged by accumulating data showing deep anatomical and functional interrelationships between both systems. In addition, the assumption that the vomeronasal system appeared as an adaptation to terrestrial life is being questioned as well. The aim of the present work is to use a comparative strategy to gain insight in our understanding of the evolution of chemical “cortex.” We have analyzed the organization of the olfactory and vomeronasal cortices of reptiles, marsupials, and placental mammals and we have compared our findings with data from other taxa in order to better understand the evolutionary history of the nasal sensory systems in vertebrates. The olfactory and vomeronsasal cortices have been re-investigated in garter snakes (Thamnophis sirtalis), short-tailed opossums (Monodelphis domestica), and rats (Rattus norvegicus) by tracing the efferents of the main and accessory olfactory bulbs using injections of neuroanatomical anterograde tracers (dextran-amines). In snakes, the medial olfactory tract is quite evident, whereas the main vomeronasal-recipient structure, the nucleus sphaericus is a folded cortical-like structure, located at the caudal edge of the amygdala. In marsupials, which are acallosal mammals, the rhinal fissure is relatively dorsal and the olfactory and vomeronasal cortices relatively expanded. Placental mammals, like marsupials, show partially overlapping olfactory and vomeronasal projections in the rostral basal telencephalon. These data raise the interesting question of how the telencephalon has been re-organized in different groups according to the biological relevance of chemical senses. PMID:21290004

  8. Cladistic analysis of olfactory and vomeronasal systems.

    Science.gov (United States)

    Ubeda-Bañon, Isabel; Pro-Sistiaga, Palma; Mohedano-Moriano, Alicia; Saiz-Sanchez, Daniel; de la Rosa-Prieto, Carlos; Gutierrez-Castellanos, Nicolás; Lanuza, Enrique; Martinez-Garcia, Fernando; Martinez-Marcos, Alino

    2011-01-01

    Most tetrapods possess two nasal organs for detecting chemicals in their environment, which are the sensory detectors of the olfactory and vomeronasal systems. The seventies' view that the olfactory system was only devoted to sense volatiles, whereas the vomeronasal system was exclusively specialized for pheromone detection was challenged by accumulating data showing deep anatomical and functional interrelationships between both systems. In addition, the assumption that the vomeronasal system appeared as an adaptation to terrestrial life is being questioned as well. The aim of the present work is to use a comparative strategy to gain insight in our understanding of the evolution of chemical "cortex." We have analyzed the organization of the olfactory and vomeronasal cortices of reptiles, marsupials, and placental mammals and we have compared our findings with data from other taxa in order to better understand the evolutionary history of the nasal sensory systems in vertebrates. The olfactory and vomeronsasal cortices have been re-investigated in garter snakes (Thamnophis sirtalis), short-tailed opossums (Monodelphis domestica), and rats (Rattus norvegicus) by tracing the efferents of the main and accessory olfactory bulbs using injections of neuroanatomical anterograde tracers (dextran-amines). In snakes, the medial olfactory tract is quite evident, whereas the main vomeronasal-recipient structure, the nucleus sphaericus is a folded cortical-like structure, located at the caudal edge of the amygdala. In marsupials, which are acallosal mammals, the rhinal fissure is relatively dorsal and the olfactory and vomeronasal cortices relatively expanded. Placental mammals, like marsupials, show partially overlapping olfactory and vomeronasal projections in the rostral basal telencephalon. These data raise the interesting question of how the telencephalon has been re-organized in different groups according to the biological relevance of chemical senses.

  9. Cladistic analysis of olfactory and vomeronasal systems

    Directory of Open Access Journals (Sweden)

    Alino eMartinez-Marcos

    2011-01-01

    Full Text Available Most tetrapods possess two nasal organs for detecting chemicals in their environment, which are the sensory detectors of the olfactory and vomeronasal systems. The seventies’ view that the olfactory system was only devoted to sense volatiles, whereas the vomeronasal system was exclusively specialized for pheromone detection was challenged by accumulating data showing deep anatomical and functional interrelationships between both systems. In addition, the assumption that the vomeronasal system appeared as an adaptation to terrestrial life is being questioned as well. The aim of the present work is to use a comparative strategy to gain insight in our understanding of the evolution of chemical cortex. We have analyzed the organization of the olfactory and vomeronasal cortices of reptiles, marsupials and placental mammals and we have compared our findings with data from other taxa in order to better understand the evolutionary history of the nasal sensory systems in vertebrates. The olfactory and vomeronsasal cortices have been re-investigated in garter snakes (Thamnophis sirtalis, short-tailed opossums (Monodelphis domestica and rats (Rattus norvegicus by tracing the efferents of the main and accessory olfactory bulbs using injections of neuroanatomical anterograde tracers (dextran-amines. In snakes, the medial olfactory tract is quite evident, whereas the main vomeronasal-recipient structure, the nucleus sphericus is a folded cortical-like structure, located at the caudal edge of the amygdala. In marsupials, which are acallosal mammals, the rhinal fissure is relatively dorsal and the olfactory and vomeronasal cortices relatively expanded. Placental mammals, like marsupials, show partially overlapping olfactory and vomeronasal projections in the rostral basal telencephalon. These data raise the interesting question of how the telencephalon has been re-organized in different groups according to the biological relevance of chemical senses.

  10. Lungfishes, like tetrapods, possess a vomeronasal system

    Directory of Open Access Journals (Sweden)

    Agustín González

    2010-09-01

    Full Text Available The vomeronasal system (VNS is an accessory olfactory system that in tetrapod vertebrates is composed of specific receptors neurons in the nasal organ and a set of centers in the forebrain that receive and relay the information consecutively towards the hypothalamus. Thus, only in tetrapods the VNS comprises a discrete vomeronasal (Jacobson’s organ, which contains receptor cells that are morphologically distinct from those of the olfactory epithelium and use different transduction mechanisms. The axons of the vomeronasal receptors in tetrapods project to the accessory olfactory bulb (AOB in the rostral telencephalon. Secondary vomeronasal connections exist through the medial amygdala to the hypothalamus. Currently, the lungfishes are considered the closest living relatives of tetrapods, based on genetic and molecular data. Here we show that the African lungfish, Protopterus dolloi, has epithelial crypts at the base of the lamellae of the olfactory epithelium that express markers of the vomeronasal receptors in tetrapods. The projections of these crypts allow us to identify an AOB on the lateral margin of the main olfactory bulb. The projections of this AOB reach a region that is topologically, hodologically, and immunohistochemically identical to the medial amygdala and could represent its homolog. Neurons of this putative medial amygdala were demonstrated to project to the lateral hypothalamus, as they do in tetrapods. All these features that lungfishes share with tetrapods indicate that lungfishes have the complete set of brain centers and connections involved in processing vomeronasal information and that these features were already present in the last common ancestor of lungfishes and tetrapods.

  11. Human vomeronasal epithelium development: An immunohistochemical overview.

    Science.gov (United States)

    Dénes, Lóránd; Pap, Zsuzsanna; Szántó, Annamária; Gergely, István; Pop, Tudor Sorin

    2015-06-01

    The vomeronasal organ (VNO) is the receptor structure of the vomeronasal system (VNS) in vertebrates. It is found bilaterally in the submucosa of the inferior part of the nasal septum. There are ongoing controversies regarding the functionality of this organ in humans. In this study we propose the immunohistochemical evaluation of changes in components of the human vomeronasal epithelium during foetal development. We used 45 foetuses of different age, which were included in three age groups. After VNO identification immunohistochemical reactions were performed using primary antibodies against the following: neuron specific enolase, calretinin, neurofilament, chromogranin, synaptophysin, cytokeratin 7, pan-cytokeratin and S100 protein. Digital slides were obtained and following colorimetric segmentation, surface area measurements were performed. The VNO was found in less than half of the studied specimens (42.2%). Neuron specific enolase and calretinin immunoexpression showed a decreasing trend with foetal age, while the other neural/neuroendocrine markers were negative in all specimens. Cytokeratin 7 expression increased with age, while Pan-Ctk had no significant variations. S100 protein immunoexpression also decreased around the VNO. The results of the present work uphold the theory of regression of the neuroepithelium that is present during initial stages of foetal development.

  12. El órgano vomeronasal humano

    OpenAIRE

    Naser G,Alfredo; Fullá O,Juan; Varas P,Mª Antonieta; Nazar S,Rodolfo

    2008-01-01

    El órgano vomeronasal (OVN) es una estructura que estudiamos alguna vez en anatomía, sin embargo su ubicación, frecuencia y función específica en humanos ha sido poco estudiada. Por este motivo se realizó una revisión bibliográfica actualizada acerca del OVN humano, enf atizando en puntos Importantes como su anatomía y relación con algunas conductas. Hoy en día es considerado como un órgano olfatorio accesorio, capaz de percibir la presencia de vomeroferinas. Estas corresponden a un grupo de ...

  13. Modelling the electrical activity of pancreatic alpha-cells based on experimental data from intact mouse islets

    DEFF Research Database (Denmark)

    Diderichsen, Paul Matthias; Gopel, S.O.

    2006-01-01

    Detailed experimental data from patch clamp experiments on pancreatic alpha-cells in intact mouse islets are used to model the electrical activity associated with glucagon secretion. Our model incorporates L- and T-type Ca2+ currents, delayed rectifying and A-type K+ currents, a voltage-gated Na......+ current, a KATP conductance, and an unspecific leak current. Tolbutamide closes KATP channels in the alpha-cell, leading to a reduction of the resting conductance from 1.1 nS to 0.4 nS. This causes the alpha-cell to depolarise from -76 mV to 33 mV. When the basal membrane potential passes the range...... between -60 and -35 mV, the alpha-cell generates action potentials. At higher voltages, the alpha-cell enters a stable depolarised state and the electrical activity ceases. The effects of tolbutamide are simulated by gradually reducing the KATP conductance (g (K,ATP) ) from 500 pS to 0 pS. When g (K...

  14. The vomeronasal organ of Lemur catta.

    Science.gov (United States)

    Smith, Timothy D; Muchlinski, Magdalena N; Bhatnagar, Kunwar P; Durham, Emily L; Bonar, Christopher J; Burrows, Anne M

    2015-02-01

    The vomeronasal organ (VNO), also known as the Jacobson's organ, is a bilateral chemosensory organ found at the base of the nasal cavity specialized for the detection of higher-molecular weight (non-volatile) chemostimuli. It has been linked to pheromone detection. The VNO has been well studied in nocturnal lemurs and lorises, but poorly studied in diurnal/cathemeral species despite the large repertoire of olfactory behaviors noted in species such as Lemur catta. Here, the VNO and associated structures were studied microanatomically in one adult female and one adult male L. catta. Traditional and immunohistochemical procedures demonstrate the VNO epithelium consists of multiple rows of sensory neurons. Immunoreactivity to Growth-associated protein 43 (GAP43) indicates the VNO is postnatally neurogenic. In volume, the VNO neuroepithelium scales similarly to palatal length compared to nocturnal strepsirrhines. Numerous taste buds present at the oral opening to the nasopalatine duct, with which the VNO communicates, provide an additional (or alternative) explanation for the flehmen behavior that has been observed in this species. The VNO of L. catta is shown to be microanatomically comparable to that of nocturnal strepsirrhines. Like nocturnal strepsirrhines, the VNO of L. catta may be functional in the reception of high-molecular weight secretions. © 2014 Wiley Periodicals, Inc.

  15. Suv39h-dependent H3K9me3 marks intact retrotransposons and silences LINE elements in mouse embryonic stem cells.

    Science.gov (United States)

    Bulut-Karslioglu, Aydan; De La Rosa-Velázquez, Inti A; Ramirez, Fidel; Barenboim, Maxim; Onishi-Seebacher, Megumi; Arand, Julia; Galán, Carmen; Winter, Georg E; Engist, Bettina; Gerle, Borbala; O'Sullivan, Roderick J; Martens, Joost H A; Walter, Jörn; Manke, Thomas; Lachner, Monika; Jenuwein, Thomas

    2014-07-17

    Heterochromatin is required to restrict aberrant expression of retrotransposons, but it remains poorly defined due to the underlying repeat-rich sequences. We dissected Suv39h-dependent histone H3 lysine 9 trimethylation (H3K9me3) by genome-wide ChIP sequencing in mouse embryonic stem cells (ESCs). Refined bioinformatic analyses of repeat subfamilies indicated selective accumulation of Suv39h-dependent H3K9me3 at interspersed repetitive elements that cover ∼5% of the ESC epigenome. The majority of the ∼8,150 intact long interspersed nuclear elements (LINEs) and endogenous retroviruses (ERVs), but only a minor fraction of the >1.8 million degenerate and truncated LINEs/ERVs, are enriched for Suv39h-dependent H3K9me3. Transcriptional repression of intact LINEs and ERVs is differentially regulated by Suv39h and other chromatin modifiers in ESCs but governed by DNA methylation in committed cells. These data provide a function for Suv39h-dependent H3K9me3 chromatin to specifically repress intact LINE elements in the ESC epigenome.

  16. Akhirin regulates the proliferation and differentiation of neural stem cells in intact and injured mouse spinal cord.

    Science.gov (United States)

    Abdulhaleem, Felemban Athary M; Song, Xiaohong; Kawano, Rie; Uezono, Naohiro; Ito, Ayako; Ahmed, Giasuddin; Hossain, Mahmud; Nakashima, Kinichi; Tanaka, Hideaki; Ohta, Kunimasa

    2015-05-01

    Although the central nervous system is considered a comparatively static tissue with limited cell turnover, cells with stem cell properties have been isolated from most neural tissues. The spinal cord ependymal cells show neural stem cell potential in vitro and in vivo in injured spinal cord. However, very little is known regarding the ependymal niche in the mouse spinal cord. We previously reported that a secreted factor, chick Akhirin, is expressed in the ciliary marginal zone of the eye, where it works as a heterophilic cell-adhesion molecule. Here, we describe a new crucial function for mouse Akhirin (M-AKH) in regulating the proliferation and differentiation of progenitors in the mouse spinal cord. During embryonic spinal cord development, M-AKH is transiently expressed in the central canal ependymal cells, which possess latent neural stem cell properties. Targeted inactivation of the AKH gene in mice causes a reduction in the size of the spinal cord and decreases BrdU incorporation in the spinal cord. Remarkably, the expression patterns of ependymal niche molecules in AKH knockout (AKH-/-) mice are different from those of AKH+/+, both in vitro and in vivo. Furthermore, we provide evidence that AKH expression in the central canal is rapidly upregulated in the injured spinal cord. Taken together, these results indicate that M-AKH plays a crucial role in mouse spinal cord formation by regulating the ependymal niche in the central canal.

  17. Establishment of NOD/SCID mouse models of human hepatocellular carcinoma via subcutaneous transplantation of histologically intact tumor tissue

    Institute of Scientific and Technical Information of China (English)

    Mingxia Yan; Hong Li; Fangyu Zhao; Lixing Zhang; Chao Ge; Ming Yao; Jinjun Li

    2013-01-01

    Hepatocellular carcinoma (HCC) is one of the most deadly human cancers,but it is very difficult to establish an animal model by using surgical specimens.In the present experiment,histologically intact fresh surgical specimens of HCC were subcutaneously transplanted in non-obese diabetic/severe combined immunodeficienccy (NOD/SCID) mice.The biological characteristics of the original and the corresponding transplanted tumors and cell lines were investigated.The results showed that 5 new animal models and 2 primary cell lines were successfully established from surgical specimens.Hematoxylin-eosin staining showed that xenografts retained major histological features of the original surgical specimens.The two new cell lines had been cultivated for 3 years and successively passaged for more than 100 passages in vitro.The morphological characteristics and biologic features of the two cell lines were genetically similar to the original tumor.The subcutaneous transplant animal models with histologically intact tumor tissue and primary cell lines could be useful for in vivo and in vitro testing of anti-cancer drugs and be ideal models to study various biologic features of HCC.

  18. Intact Pericellular Matrix of Articular Cartilage Is Required for Unactivated Discoidin Domain Receptor 2 in the Mouse Model

    Science.gov (United States)

    Xu, Lin; Polur, Ilona; Servais, Jacqueline M.; Hsieh, Sirena; Lee, Peter L.; Goldring, Mary B.; Li, Yefu

    2011-01-01

    Increased expression of the discoidin domain receptor 2 (DDR2) results from its interaction with collagen type II. This induces expression of matrix metalloproteinase (MMP)-13, leading to osteoarthritis (OA). To investigate the impact of the pericellular matrix of chondrocytes on DDR2, we generated a mouse model with inducible overexpression of DDR2 in cartilage. Conditional overexpression of DDR2 in mature mouse articular cartilage was controlled via the cartilage oligomeric matrix protein promoter using the Tet-Off-inducible system. Doxycycline was withdrawn at 1 month of age, and knee joints were examined at 2, 3, and 4 months of age. Microsurgery was performed on 3-month-old transgenic mice overexpressing DDR2 to destabilize the medial meniscus, and serial paraffin sections were examined at 2, 4, 8, and 12 weeks after surgery. DDR2 expression increased in the knee joints of transgenic mice. However, the increased DDR2 did not induce MMP-13 expression. No OA-like changes were observed in the transgenic mice at the age of 4 months. When transgenic mice were subjected to destabilizing of the medial meniscus, we observed accelerated progression to OA, which was associated with DDR2 activation. Therefore, conditionally overexpressing DDR2 in the mature articular cartilage of mouse knee joints requires activation to induce OA, and altered biomechanical stress can accelerate the onset of cartilage loss and progression to OA in transgenic mice. PMID:21855682

  19. Intact pericellular matrix of articular cartilage is required for unactivated discoidin domain receptor 2 in the mouse model.

    Science.gov (United States)

    Xu, Lin; Polur, Ilona; Servais, Jacqueline M; Hsieh, Sirena; Lee, Peter L; Goldring, Mary B; Li, Yefu

    2011-09-01

    Increased expression of the discoidin domain receptor 2 (DDR2) results from its interaction with collagen type II. This induces expression of matrix metalloproteinase (MMP)-13, leading to osteoarthritis (OA). To investigate the impact of the pericellular matrix of chondrocytes on DDR2, we generated a mouse model with inducible overexpression of DDR2 in cartilage. Conditional overexpression of DDR2 in mature mouse articular cartilage was controlled via the cartilage oligomeric matrix protein promoter using the Tet-Off-inducible system. Doxycycline was withdrawn at 1 month of age, and knee joints were examined at 2, 3, and 4 months of age. Microsurgery was performed on 3-month-old transgenic mice overexpressing DDR2 to destabilize the medial meniscus, and serial paraffin sections were examined at 2, 4, 8, and 12 weeks after surgery. DDR2 expression increased in the knee joints of transgenic mice. However, the increased DDR2 did not induce MMP-13 expression. No OA-like changes were observed in the transgenic mice at the age of 4 months. When transgenic mice were subjected to destabilizing of the medial meniscus, we observed accelerated progression to OA, which was associated with DDR2 activation. Therefore, conditionally overexpressing DDR2 in the mature articular cartilage of mouse knee joints requires activation to induce OA, and altered biomechanical stress can accelerate the onset of cartilage loss and progression to OA in transgenic mice.

  20. Sustained Administration of β-cell Mitogens to Intact Mouse Islets Ex Vivo Using Biodegradable Poly(lactic-co-glycolic acid) Microspheres.

    Science.gov (United States)

    Pasek, Raymond C; Kavanaugh, Taylor E; Duvall, Craig L; Gannon, Maureen A

    2016-11-05

    The development of biomaterials has significantly increased the potential for targeted drug delivery to a variety of cell and tissue types, including the pancreatic β-cells. In addition, biomaterial particles, hydrogels, and scaffolds also provide a unique opportunity to administer sustained, controllable drug delivery to β-cells in culture and in transplanted tissue models. These technologies allow the study of candidate β-cell proliferation factors using intact islets and a translationally relevant system. Moreover, determining the effectiveness and feasibility of candidate factors for stimulating β-cell proliferation in a culture system is critical before moving forward to in vivo models. Herein, we describe a method to co-culture intact mouse islets with biodegradable compound of interest (COI)-loaded poly(lactic-co-glycolic acid) (PLGA) microspheres for the purpose of assessing the effects of sustained in situ release of mitogenic factors on β-cell proliferation. This technique describes in detail how to generate PLGA microspheres containing a desired cargo using commercially available reagents. While the described technique uses recombinant human Connective tissue growth factor (rhCTGF) as an example, a wide variety of COI could readily be used. Additionally, this method utilizes 96-well plates to minimize the amount of reagents necessary to assess β-cell proliferation. This protocol can be readily adapted to use alternative biomaterials and other endocrine cell characteristics such as cell survival and differentiation status.

  1. Genomic variation in the vomeronasal receptor gene repertoires of inbred mice

    Directory of Open Access Journals (Sweden)

    Wynn Elizabeth H

    2012-08-01

    Full Text Available Abstract Background Vomeronasal receptors (VRs, expressed in sensory neurons of the vomeronasal organ, are thought to bind pheromones and mediate innate behaviours. The mouse reference genome has over 360 functional VRs arranged in highly homologous clusters, but the vast majority are of unknown function. Differences in these receptors within and between closely related species of mice are likely to underpin a range of behavioural responses. To investigate these differences, we interrogated the VR gene repertoire from 17 inbred strains of mice using massively parallel sequencing. Results Approximately half of the 6222 VR genes that we investigated could be successfully resolved, and those that were unambiguously mapped resulted in an extremely accurate dataset. Collectively VRs have over twice the coding sequence variation of the genome average; but we identify striking non-random distribution of these variants within and between genes, clusters, clades and functional classes of VRs. We show that functional VR gene repertoires differ considerably between different Mus subspecies and species, suggesting these receptors may play a role in mediating behavioural adaptations. Finally, we provide evidence that widely-used, highly inbred laboratory-derived strains have a greatly reduced, but not entirely redundant capacity for differential pheromone-mediated behaviours. Conclusions Together our results suggest that the unusually variable VR repertoires of mice have a significant role in encoding differences in olfactory-mediated responses and behaviours. Our dataset has expanded over nine fold the known number of mouse VR alleles, and will enable mechanistic analyses into the genetics of innate behavioural differences in mice.

  2. Optimal clearance of Sporothrix schenckii requires an intact Th17 response in a mouse model of systemic infection.

    Science.gov (United States)

    Ferreira, Lucas Souza; Gonçalves, Amanda Costa; Portuondo, Deivys Leandro; Maia, Danielle Cardoso Geraldo; Placeres, Marisa Campos Polesi; Batista-Duharte, Alexander; Carlos, Iracilda Zeppone

    2015-08-01

    The discovery of Th17 cells, along with many other Th cell subsets in the recent years, has expanded the Th1/Th2 paradigm that had persisted since its proposition by Mosmann in 1986. Defined by the characteristic expression of the transcription factor retinoic-related orphan receptor γt (RORγt) and production of IL-17A (IL-17), Th17 cells are powerful inducers of tissue inflammation with a recognized role against extracellular bacteria and fungi. Despite this, the interest in their study came from the pivotal role they play in the development and maintenance of major chronic inflammatory conditions such as multiple sclerosis, rheumatoid arthritis and Crohn's disease, hence they have been the target of promising new anti-Th17 therapies. Accordingly, the identification of opportunistic pathogens whose clearance relies on the Th17 response is of huge prophylactic importance. As shown here for the first time, this applies to Sporothrix schenckii, a thermo-dimorphic fungus and the causative agent of sporotrichosis. Our results show that both Th17 and Th1/Th17 mixed cells are developed during the S. schenckii systemic mice infection, which also leads to augmented production of IL-17 and IL-22. Also, by using an antibody-mediated IL-23 depletion model, we further demonstrate that optimal fungal clearance, but not survival, depends on an intact Th17 response.

  3. Variations in Local Calcium Signaling in Adjacent Cardiac Myocytes of the Intact Mouse Heart Detected with Two-Dimensional Confocal Microscopy

    Directory of Open Access Journals (Sweden)

    Karin P Hammer

    2015-01-01

    Full Text Available Dyssynchronous local Ca release within individual cardiac myocytes has been linked to cellular contractile dysfunction. Differences in Ca kinetics in adjacent cells may also provide a substrate for inefficient contraction and arrhythmias. In a new approach we quantify variation in local Ca transients between adjacent myocytes in the whole heart.Langendorff-perfused mouse hearts were loaded with Fluo-8 AM to detect Ca and Di-4-ANEPPS to visualize cell membranes. A spinning disc confocal microscope with a fast camera allowed us to record Ca signals within an area of 465 µm by 315 µm with an acquisition speed of 55 fps. Images from multiple transients recorded at steady state were registered to their time point in the cardiac cycle to restore averaged local Ca transients with a higher temporal resolution. Local Ca transients within and between adjacent myocytes were compared with regard to amplitude, time to peak and decay at steady state stimulation (250 ms cycle length.Image registration from multiple sequential Ca transients allowed reconstruction of high temporal resolution (2.4 ±1.3ms local CaT in 2D image sets (N= 4 hearts, n= 8 regions. During steady state stimulation, spatial Ca gradients were homogeneous within cells in both directions and independent of distance between measured points. Variation in CaT amplitudes was similar across the short and the long side of neighboring cells. Variations in TAU and TTP were similar in both directions. Isoproterenol enhanced the CaT but not the overall pattern of spatial heterogeneities.Here we detected and analyzed local Ca signals in intact mouse hearts with high temporal and spatial resolution, taking into account 2D arrangement of the cells. We observed significant differences in the variation of CaT amplitude along the long and short axis of cardiac myocytes. Variations of Ca signals between neighboring cells may contribute to the substrate of cardiac remodeling.

  4. Developmental studies on the rat vomeronasal organ: vascular pattern and neuroepithelial differentiation. I. Light microscopy.

    Science.gov (United States)

    Szabó, K; Mendoza, A S

    1988-04-01

    The origin and the developmental sequence of the rat vomeronasal organ and its vascular supply are followed by means of India ink injection in serial sections of celloidin-embedded embryos from the eleventh day of gestation up to birth. The anlage of the vomeronasal organ has been established by the twelfth day of gestation (E 12). It appears as a shallow longitudinal impression of the medial wall of the nasal pit. At day E 14, it separates from the epithelium of the primary nasal cavity, forming a tube. The lumen of the organ remains continuous with the nasal cavity frontally, but ends blindly at the edge of the primary palate dorsally. From day E 16 to E 18 the lateral surface of the tubular vomeronasal organ invaginates toward the lumen forming a wide longitudinal furrow. The lumen is bordered by the developing neuroepithelium and receptor-free epithelium by this time. The vomeronasal organ receives a separate arterial blood supply arising from septal tributaries of the olfactory artery, a branch of the anterior cerebral artery from the earliest stage of development. Blood from the vomeronasal complex is collected in the vomeronasal vein lying in the longitudinal furrow next to the receptor-free epithelium. The typical vascular pattern of the vomeronasal organ is established by the eighteenth day of gestation. At this time, the first capillary loops appear within the neuroepithelium and the vomeronasal vein can already be seen to extend along the long axis of the organ.

  5. Evolution and origin of vomeronasal-type odorant receptor gene repertoire in fishes

    Directory of Open Access Journals (Sweden)

    Nishida Mutsumi

    2006-10-01

    Full Text Available Abstract Background In teleost fishes that lack a vomeronasal organ, both main odorant receptors (ORs and vomeronasal receptors family 2 (V2Rs are expressed in the olfactory epithelium, and used for perception of water-soluble chemicals. In zebrafish, it is known that both ORs and V2Rs formed multigene families of about a hundred copies. Whereas the contribution of V2Rs in zebrafish to olfaction has been found to be substantially large, the composition and structure of the V2R gene family in other fishes are poorly known, compared with the OR gene family. Results To understand the evolutionary dynamics of V2R genes in fishes, V2R sequences in zebrafish, medaka, fugu, and spotted green pufferfish were identified from their draft genome sequences. There were remarkable differences in the number of intact V2R genes in different species. Most V2R genes in these fishes were tightly clustered in one or two specific chromosomal regions. Phylogenetic analysis revealed that the fish V2R family could be subdivided into 16 subfamilies that had diverged before the separation of the four fishes. Genes in two subfamilies in zebrafish and another subfamily in medaka increased in their number independently, suggesting species-specific evolution in olfaction. Interestingly, the arrangements of V2R genes in the gene clusters were highly conserved among species in the subfamily level. A genomic region of tetrapods corresponding to the region in fishes that contains the V2R cluster was found to have no V2R gene in any species. Conclusion Our results have indicated that the evolutionary dynamics of fish V2Rs are characterized by rapid gene turnover and lineage-specific phylogenetic clustering. In addition, the present phylogenetic and comparative genome analyses have shown that the fish V2Rs have expanded after the divergence between teleost and tetrapod lineages. The present identification of the entire V2R repertoire in fishes would provide useful foundation to

  6. Prey detection by vomeronasal chemoreception in a plethodontid salamander.

    Science.gov (United States)

    Placyk, John S; Graves, Brent M

    2002-05-01

    While chemoreception is involved in a wide variety of salamander behaviors, the chemosensory system that mediates specific behaviors is rarely known. We investigated the role of the vomeronasal system (VNS) in foraging behavior of the red-backed salamander (Plethodon cinereus) by manipulating salamanders' abilities to detect nonvolatile chemical cues emitted by potential prey. Subjects received one of three treatments: (1) impaired vomeronasal system, (2) sham manipulation, and (3) no manipulation. The role of the VNS in mediating foraging on motile prey (Drosophila melanogaster) was investigated under three light conditions (bright, dim, dark). Salamanders with impaired VNSs foraged less efficiently than either of the other experimental groups by displaying the longest latency to attack and the lowest rate of prey capture, especially in the absence of visual cues. A second experiment utilized freshly killed prey to determine whether the VNS takes on added importance in the absence of visual or tactile cues associated with moving prey. Animals with impaired VNSs showed a decreased foraging efficiency on stationary prey under both dark and light conditions. In addition, a mark-recapture study of VNS-impaired and sham salamanders in the field also indicated that salamanders with impaired VNSs consumed fewer stationary prey compared to shams. The study indicates that the VNS plays a substantial role in the foraging behavior of the plethodontid salamander, P. cinereus.

  7. Sexual and seasonal differences in the vomeronasal epithelium of the red-backed salamander (Plethodon cinereus).

    Science.gov (United States)

    Dawley, E M; Crowder, J

    1995-08-28

    Sexually dimorphic behaviors often are associated with sexually dimorphic neural structures. Perinatal hormonal levels influence structural sexual dimorphism, and seasonal structural changes also can be the result of seasonal hormonal fluctuations. We compared the volume of vomeronasal organs of male and female red-backed salamanders (Plethodon cinereus) throughout the year. Odorants are delivered to vomeronasal receptors through nasolabial grooves when salamanders touch the bases of these grooves to objects (nose tapping). Males may locate and identify potential mates through nose tapping during the prolonged breeding season that lasts from October through May. We compared vomeronasal organ data through multiple regression by using total body size, sex, and season as variables that may influence vomeronasal organ volume. Gonads also were examined as an indicator of reproductive status. Total body size and sex significantly affect vomeronasal organ volume; as body size increases, so does vomeronasal organ volume, and males have significantly larger vomeronasal organs than females at all times of the year. During the summer, both males and females have larger vomeronasal organs than at any other time of the year. Summer also is a period of intense foraging and the initiation of a new cycle of gametogenesis. Previous studies of circulating hormone levels in amphibians have shown that the initial stages of gametogenesis correspond to a period of low estradiol and testosterone levels but high gonadotropin-releasing hormone levels. The functional significance of sexual and seasonal differences in the vomeronasal organs of P. cinereus may relate to the neurogenesis of specialized receptors for courtship and mating.

  8. The role of the vomeronasal organ in rattlesnake (Crotalus viridis oreganus) predatory behavior.

    Science.gov (United States)

    Alving, W R; Kardong, K V

    1996-01-01

    During predatory behavior, rattlesnakes depend primarily upon thermal and visual cues to initially aim a strike. However, it has been hypothesized that prey-related odors sensed by the vomeronasal system act as releasing stimuli of the strike and that such vomodors are primary stimuli during poststrike trailing and swallowing of the envenomated rodent. To test this, northern Pacific rattlesnakes were rendered avomic by bilateral lesions of the vomeronasal nerves, and their vomic and avomic predatory behaviors were compared. Avomic rattlesnakes exhibited fewer strikes and complete elimination of trailing and swallowing behavior. These results support the hypothesis that vomodors sensed via the vomeronasal organ are capable of acting as releasing stimuli of selected rattlesnake predatory behaviors. Sensory input via the vomeronasal organ is important during prestrike/strike behavior, and it is a major route of sensory input during poststrike trailing and ingestion of envenomated prey.

  9. Fate and Development of Human Vomeronasal Organ - A Microscopic Fetal Study.

    Science.gov (United States)

    Vasuki, A K Manicka; Fenn, T K Aleyemma; Devi, M Nirmala; Hebzibah, T Deborah Joy; Jamuna, M; Sundaram, K Kalyana

    2016-03-01

    The existence of Vomeronasal organ in human is a controversial subject. Presence of Vomeronasal organ and its structure was not reported in standard text books. The presence of Vomeronasal organ in fetal life is doubtful. Hence identification of the organ by histological examination was planned. A study was conducted on resected specimens of nasal septum obtained from 45 spontaneously aborted fetuses from Obstetrics and Gynaecology department, PSG Institute of Medical Sciences and Research, Coimbatore, after ethical clearance. The histological structure of Vomeronasal organ was observed from 11 weeks old fetus. The epithelial lining of the organ, presence of cilia, presence of lamina propria, acini and the blood vessel and the types of cells were observed. The organ was lined by pseudostratified columnar epithelium. The organ showed Lamina propria with serous acini from 18 weeks fetus. Vomeronasal duct opening into the nasal cavity and three types of cells were observed in 28 weeks fetus. Knowledge about the persistence of Vomeronasal organ in fetuses and its structure need to be known. The organ may be found as a putative pit posterior to anterior nasal spine. The organ may be damaged in nasal septal surgeries and nasal endoscopic procedures. The organ may not be seen on gross examination in all human fetuses and cadavers.

  10. Neurogenesis in the vomeronasal epithelium of adult garter snakes: 3. Use of /sup 3/H-thymidine autoradiography to trace the genesis and migration of bipolar neurons

    Energy Technology Data Exchange (ETDEWEB)

    Wang, R.T.; Halpern, M.

    1988-10-01

    Use of 3H-thymidine autoradiography and unilateral vomeronasal (VN) axotomy has permitted us to demonstrate directly the existence of VN stem cells in the adult garter snake and to trace continuous bipolar neuron development and migration in the normal VN and deafferentated VN epithelium in the same animal. The vomeronasal epithelium and olfactory epithelium of adult garter snakes are both capable of incorporating 3H-thymidine. In the sensory epithelium of the vomeronasal organ, 3H-thymidine-labeled cells were initially restricted to the base of the undifferentiated cell layer in animals surviving 1 day following 3H-thymidine injection. With increasing survival time, labeled cells progressively migrated vertically within the receptor cell column toward the apex of the bipolar neuron layer. In both the normal and denervated VN epithelium, labeled cells were observed through the 56 days of postoperative survival. In the normal epithelium, labeled cells were always located within the matrix of the intact receptor cell columns. However, labeled cells of the denervated epithelium were always located at the apical front of the newly formed cell mass following depletion of the original neuronal cell population. In addition, at postoperative days 28 and 56, labeled cells of the denervated VN epithelium achieved neuronal differentiation and maturation by migrating much farther away from the base of the receptor cell column than the labeled cells on the normal, unoperated contralateral side. This study directly demonstrates that basal cells initially incorporating 3H-thymidine are indeed stem cells of the VN epithelium in adult garter snakes.

  11. Male pheromone protein components activate female vomeronasal neurons in the salamander Plethodon shermani

    Directory of Open Access Journals (Sweden)

    Feldhoff Pamela W

    2006-03-01

    Full Text Available Abstract Background The mental gland pheromone of male Plethodon salamanders contains two main protein components: a 22 kDa protein named Plethodon Receptivity Factor (PRF and a 7 kDa protein named Plethodon Modulating Factor (PMF, respectively. Each protein component individually has opposing effects on female courtship behavior, with PRF shortening and PMF lengthening courtship. In this study, we test the hypothesis that PRF or PMF individually activate vomeronasal neurons. The agmatine-uptake technique was used to visualize chemosensory neurons that were activated by each protein component individually. Results Vomeronasal neurons exposed to agmatine in saline did not demonstrate significant labeling. However, a population of vomeronasal neurons was labeled following exposure to either PRF or PMF. When expressed as a percent of control level labeled cells, PRF labeled more neurons than did PMF. These percentages for PRF and PMF, added together, parallel the percentage of labeled vomeronasal neurons when females are exposed to the whole pheromone. Conclusion This study suggests that two specific populations of female vomeronasal neurons are responsible for responding to each of the two components of the male pheromone mixture. These two neural populations, therefore, could express different receptors which, in turn, transmit different information to the brain, thus accounting for the different female behavior elicited by each pheromone component.

  12. [Comparative anatomic studies of the vomeronasal complex and the rostral palate of various mammals].

    Science.gov (United States)

    Wöhrmann-Repenning, A

    1984-01-01

    The structures of the rostral palate in regard to the vomeronasal complex of different species of mammals were studied. In all cases, we find a very interesting system of furrows which preserves a connection between the nasopalatine ducts and the preoral surroundings. For rodents, lagomorphs, Solenodon, Setifer, and Echinops, we find a special situation in this part of the palate. Here the incisors are not separated by a diastema nor the oral openings of the nasopalatine ducts are overgrown by a bipartite caudal branch of the rhinarium. The results of the anatomic studies of the vomeronasal complex and the rostral palate of the mammals investigated are discussed: First of all, some elements of the vomeronasal complex needed to be analysed in regard to structure and nomenclature, specifically the cartilago paraseptalis with its outer bar, the cartilago ductus nasopalatini and the cartilago palatina. Because of 2 criterions, the vomeronasal complex could be classified as either primitive or progressive. We find a primitive one in Didelphis, Tupaia, Solenodon, Oryctolagus, and all rodents. In contrast, the other insectivores studied and all primates show progressive structures at their vomeronasal complex. Finally, conclusions in regard to the function of the organs of Jacobson are derived from these studies. The significance of the "flehmen" mechanism for the functioning of the organs is questioned.

  13. The structure of the nasal chemosensory system in squamate reptiles. 2. Lubricatory capacity of the vomeronasal organ

    Indian Academy of Sciences (India)

    Susan J Rehorek; Bruce T Firth; Mark N Hutchinson

    2000-06-01

    The vomeronasal organ is a poorly understood accessory olfactory organ, present in many tetrapods. In mammals, amphibians and lepidosaurian reptiles, it is an encapsulated structure with a central, fluid-filled lumen. The morphology of the lubricatory system of the vomeronasal organ (the source of this fluid) varies among classes, being either intrinsic (mammalian and caecilian amphibian vomeronasal glands) or extrinsic (anuran and urodele nasal glands). In the few squamate reptiles thus far examined, there are no submucosal vomeronasal glands. In this study, we examined the vomeronasal organs of several species of Australian squamates using histological, histochemical and ultrastructural techniques, with the goal of determining the morphology of the lubricatory system in the vomeronasal organ. Histochemically, the fluid within the vomeronasal organ of all squamates is mucoserous, though it is uncertain whether mucous and serous constituents constitute separate components. The vomeronasal organ produces few secretory granules intrinsically, implying an extrinsic source for the luminal fluid. Of three possible candidates, the Harderian gland is the most likely extrinsic source of this secretion.

  14. TRICK or TRP? What Trpc2-/- Mice Tell Us about Vomeronasal Organ Mediated Innate Behaviors

    Directory of Open Access Journals (Sweden)

    C. Ron eYu

    2015-06-01

    Full Text Available The vomeronasal organ (VNO plays an important role in mediating semiochemical communications and social behaviors in terrestrial species. Genetic knockout of individual components in the signaling pathways have been used to probe vomeronasal functions, and have provided much insights into how the VNO orchestrates innate behaviors. However, all data do not agree. In particular, knocking out Trpc2, a member of the TRP family of non-selective cationic channel thought to be the main transduction channel in the VNO, results in a number of fascinating behavioral phenotypes that have not been observed in other animals whose vomeronasal function is disrupted. Recent studies have identified signaling pathways that operate in parallel of Trpc2, raising the possibility that Trpc2 mutant animals may display neomorphic behaviors. In this article, I provide a critical analysis of emerging evidence to reconcile the discrepancies and discuss their implications.

  15. Contrasted evolution of the vomeronasal receptor repertoires in mammals and squamate reptiles.

    Science.gov (United States)

    Brykczynska, Urszula; Tzika, Athanasia C; Rodriguez, Ivan; Milinkovitch, Michel C

    2013-01-01

    The vomeronasal organ (VNO) is an olfactory structure that detects pheromones and environmental cues. It consists of sensory neurons that express evolutionary unrelated groups of transmembrane chemoreceptors. The predominant V1R and V2R receptor repertoires are believed to detect airborne and water-soluble molecules, respectively. It has been suggested that the shift in habitat of early tetrapods from water to land is reflected by an increase in the ratio of V1R/V2R genes. Snakes, which have a very large VNO associated with a sophisticated tongue delivery system, are missing from this analysis. Here, we use RNA-seq and RNA in situ hybridization to study the diversity, evolution, and expression pattern of the corn snake vomeronasal receptor repertoires. Our analyses indicate that snakes and lizards retain an extremely limited number of V1R genes but exhibit a large number of V2R genes, including multiple lineages of reptile-specific and snake-specific expansions. We finally show that the peculiar bigenic pattern of V2R vomeronasal receptor gene transcription observed in mammals is conserved in squamate reptiles, hinting at an important but unknown functional role played by this expression strategy. Our results do not support the hypothesis that the shift to a vomeronasal receptor repertoire dominated by V1Rs in mammals reflects the evolutionary transition of early tetrapods from water to land. This study sheds light on the evolutionary dynamics of the vomeronasal receptor families in vertebrates and reveals how mammals and squamates differentially adapted the same ancestral vomeronasal repertoire to succeed in a terrestrial environment.

  16. The molecular evolutionary dynamics of the vomeronasal receptor (class 1) genes in primates: a gene family on the verge of a functional breakdown.

    Science.gov (United States)

    Yoder, Anne D; Larsen, Peter A

    2014-01-01

    Olfaction plays a critical role in both survival of the individual and in the propagation of species. Studies from across the mammalian clade have found a remarkable correlation between organismal lifestyle and molecular evolutionary properties of receptor genes in both the main olfactory system (MOS) and the vomeronasal system (VNS). When a large proportion of intact (and putatively functional) copies is observed, the inference is made that a particular mode of chemoreception is critical for an organism's fit to its environment and is thus under strong positive selection. Conversely, when the receptors in question show a disproportionately large number of pseudogene copies, this contraction is interpreted as evidence of relaxed selection potentially leading to gene family extinction. Notably, it appears that a risk factor for gene family extinction is a high rate of nonsynonymous substitution. A survey of intact vs. pseudogene copies among primate vomeronasal receptor Class one genes (V1Rs) appears to substantiate this hypothesis. Molecular evolutionary complexities in the V1R gene family combine rapid rates of gene duplication, gene conversion, lineage-specific expansions, deletions, and/or pseudogenization. An intricate mix of phylogenetic footprints and current adaptive landscapes have left their mark on primate V1Rs suggesting that the primate clade offers an ideal model system for exploring the molecular evolutionary and functional properties of the VNS of mammals. Primate V1Rs tell a story of ancestral function and divergent selection as species have moved into ever diversifying adaptive regimes. The sensitivity to functional collapse in these genes, consequent to their precariously high rates of nonsynonymous substitution, confer a remarkable capacity to reveal the lifestyles of the genomes that they presently occupy as well as those of their ancestors.

  17. The myelin basic protein-specific T cell repertoire in (transgenic) Lewis rat/SCID mouse chimeras: preferential V beta 8.2 T cell receptor usage depends on an intact Lewis thymic microenvironment.

    Science.gov (United States)

    Kääb, G; Brandl, G; Marx, A; Wekerle, H; Bradl, M

    1996-05-01

    In the Lewis rat, myelin basic protein (MBP)-specific, encephalitogenic T cells preferentially recognize sequence 68-88, and use the V beta 8.2 gene to encode their T cell receptors. To analyze the structural prerequisites for the development of the MBP-specific T cell repertoire, we reconstituted severe-combined immunodeficient (SCID) mice with fetal (embryonic day 15-16) Lewis rat lymphoid tissue, and then isolated MBP-specific T cell lines from the adult chimeras after immunization. Two types of chimera were constructed: SCID mice reconstituted with rat fetal liver cells only, allowing T cell maturation within a chimeric SCID thymus consisting of mouse thymic epithelium and rat interdigitating dendritic cells, and SCID mice reconstituted with rat fetal liver cells and rat fetal thymus grafts, allowing T cell maturation within the chimeric SCID and the intact Lewis rat thymic microenvironment. Without exception, the T cell lines isolated from MBP-immunized SCID chimeras were restricted by MHC class II of the Lewis rat (RT1.B1), and none by I-Ad of the SCID mouse. Most of the T cell lines recognized the immunodominant MBP epitope 68-88. In striking contrast to intact Lewis rats, in SCID mice reconstituted by rat fetal liver only, MBP-specific T cell clones used a seemingly random repertoire of V beta genes without a bias for V beta 8.2. In chimeras containing fetal Lewis liver plus fetal thymus grafted under the kidney capsule, however, dominant utilization of V beta 8.2 was restored. The migration of liver-derived stem cells through rat thymus grafts was documented by combining fetal tissues from wild-type and transgenic Lewis rats. The results confirm that the recognition of the immunodominant epitope 68-88 by MBP-specific encephalitogenic T cells is a genetically determined feature of the Lewis rat T cell repertoire. They further suggest that the formation of the repertoire requires T cell differentiation in a syngeneic thymic microenvironment.

  18. Evolutionary deterioration of the vomeronasal pheromone transduction pathway in catarrhine primates

    OpenAIRE

    2003-01-01

    Pheromones are water-soluble chemicals released and sensed by individuals of the same species to elicit social and reproductive behaviors or physiological changes; they are perceived primarily by the vomeronasal organ (VNO) in terrestrial vertebrates. Humans and some related primates possess only vestigial VNOs and have no or significantly reduced ability to detect pheromones, a phenomenon not well understood at the molecular level. Here we show that genes encoding the...

  19. Human Pheromone Detection by the Vomeronasal Organ: Unnecessary for Mate Selection?

    OpenAIRE

    2009-01-01

    Recently, Foltan and Sedy proposed a hypothesis stating that the adult human VNO is integral to the prevention of inappropriate mate selection. In this commentary, we address the authors’ assumption that humans have a functional VNO, that pheromones are detected exclusively by the VNO, and that human pheromones are responsible for negative stimuli during mate selection. After examining the published literature on human vomeronasal function, we argue that their hypothesis is critically flawed....

  20. The shrinking anthropoid nose, the human vomeronasal organ, and the language of anatomical reduction.

    Science.gov (United States)

    Smith, Timothy D; Laitman, Jeffrey T; Bhatnagar, Kunwar P

    2014-11-01

    Humans and most of our closest extant relatives, the anthropoids, are notable for their reduced "snout." The striking reduction in facial projection is only a superficial similarity. All anthropoids, including those with long faces (e.g., baboons), have lost numerous internal projections (turbinals) and spaces (recesses). In sum, this equates to the loss of certain regions of olfactory mucosa in anthropoids. In addition, an accessory olfactory organ, the vomeronasal organ, is non-functional or even absent in all catarrhine primates (humans, apes, monkeys). In this commentary, we revisit the concept of anatomical reductions as it pertains to the anthropoid nasal region. Certain nasal structures and spaces in anthropoids exhibit well-known attributes of other known vestiges, such as variability in form or number. The cupular recess (a vestige of the olfactory recess) and some rudimentary ethmoturbinals constitute reduced structures that presumably were fully functional in our ancestors. Humans and at least some apes retain a vestige that is bereft of chemosensory function (while in catarrhine monkeys it is completely absent). However, the function of the vomeronasal system also includes prenatal roles, which may be common to most or all mammals. Notably, neurons migrate to the brain along vomeronasal and terminal nerve axons during embryogenesis. The time-specific role of the VNO raises the possibility that our concept of functional reduction is too static. The vomeronasal system of humans and other catarrhine primates appears to qualify as a "chronological" vestige, one which fulfills part of its function during ontogeny, and then becomes lost or vestigial.

  1. Evolutionary deterioration of the vomeronasal pheromone transduction pathway in catarrhine primates.

    Science.gov (United States)

    Zhang, Jianzhi; Webb, David M

    2003-07-08

    Pheromones are water-soluble chemicals released and sensed by individuals of the same species to elicit social and reproductive behaviors or physiological changes; they are perceived primarily by the vomeronasal organ (VNO) in terrestrial vertebrates. Humans and some related primates possess only vestigial VNOs and have no or significantly reduced ability to detect pheromones, a phenomenon not well understood at the molecular level. Here we show that genes encoding the TRP2 ion channel and V1R pheromone receptors, two components of the vomeronasal pheromone signal transduction pathway, have been impaired and removed from functional constraints since shortly before the separation of hominoids and Old World monkeys approximately 23 million years ago, and that the random inactivation of pheromone receptor genes is an ongoing process even in present-day humans. The phylogenetic distribution of vomeronasal pheromone insensitivity is concordant with those of conspicuous female sexual swelling and male trichromatic color vision, suggesting that a vision-based signaling-sensory mechanism may have in part replaced the VNO-mediated chemical-based system in the social/reproductive activities of hominoids and Old World monkeys (catarrhines).

  2. Scanning electron microscopic studies of the surface morphology of the vomeronasal epithelium and olfactory epithelium of garter snakes.

    Science.gov (United States)

    Wang, R T; Halpern, M

    1980-04-01

    Fixed vomeronasal and olfactory epithelia from normal adult garter snakes were microdissected, fractured, and examined with a scanning electron microscope. The method permits a detailed comparative study of the structural organization and morphological characteristics of the constituent cells of the vomeronasal and olfactory epithelia. Despite similarities in the nomenclature of the constituent cells in both epithelia, significant differences exist in their surface morphology. A unique columnar structure composed of non-neuronal elements is present in the vomeronasal epithelium. These columns house the bioplar neurons and undifferentiated cells. Such a columnar organization is absent in the olfactory epithelium. In vomeronasal epithelium the bipolar neurons possess microvillous terminals at their dendritic tips, while the dendritic tips of the bipolar neurons of the olfactory epithelium possess cilia. Vomeronasal supporting cells are covered with microvilli, while olfactory supporting cells are covered with cytoplasmic protuberances in addition to the microvilli. In the vomeronasal epithelium the pear-shaped neurons have a grossly smooth surface and are organized into clusters, while in the olfactory epithelium the elliptical bipolar neurons are spinous, aligned side-by-side and interdigitate. The basal (undifferentiated) cell layer in the vomeronasal epithelium has a high packing density and is composed of several layers of irregularly shaped cells. In the olfactory epithelium the basal cell layer is loosely organized and composed of a single layer of oval cells. This information on the three-dimensional cell structure of both epithelia provides a basis for experimental observations on changes in morphology of the bipolar neurons during genesis, development, maturation, degeneration, and regeneration in postnatal, adult animals.

  3. Non-intact zona improves development of murine preimplantation ...

    African Journals Online (AJOL)

    ajl5

    2012-09-25

    Sep 25, 2012 ... African Journal of Biotechnology Vol. 11(77), pp. ... Key words: Mouse, non-intact zona embryos, adenovirus vector with green fluorescent protein (pAd-GFP), embryos ..... informational molecule, could be lysised or its function.

  4. Efferent connections of the "olfactostriatum": a specialized vomeronasal structure within the basal ganglia of snakes.

    Science.gov (United States)

    Martinez-Marcos, Alino; Ubeda-Bañon, Isabel; Lanuza, Enrique; Halpern, Mimi

    2005-05-01

    The olfactostriatum is a portion of the basal ganglia of snakes that receives substantial vomeronasal afferents through projections from the nucleus sphericus. In a preceding article, the olfactostriatum of garter snakes (Thamnophis sirtalis) was characterized on the basis of chemoarchitecture (distribution of serotonin, neuropeptide Y and tyrosine hydroxylase) and pattern of afferent connections [Martinez-Marcos, A., Ubeda-Banon, I., Lanuza, E., Halpern, M., 2005. Chemoarchitecture and afferent connections of the "olfactostriatum": a specialized vomeronasal structure within the basal ganglia of snakes. J. Chem. Neuroanat. 29, 49-69]. In the present study, its efferent connections have been investigated. The olfactostriatum projects to the main and accessory olfactory bulbs, lateral cortex, septal complex, ventral pallidum, external, ventral anterior and dorsolateral amygdalae, bed nucleus of the stria terminalis, preoptic area, lateral posterior hypothalamic nucleus, ventral tegmental area, substantia nigra and raphe nuclei. Tracer injections in the nucleus accumbens proper, a structure closely associated with the olfactostriatum, result in a similar pattern of efferent connections with the exception of those reaching the main and accessory olfactory bulbs, lateral cortex, external, ventral anterior and dorsolateral amygdalae and bed nucleus of the stria terminalis. These data, therefore, help to characterize the olfactostriatum, an apparently specialized area of the nucleus accumbens. Double labeling experiments after tracer injections in the nucleus sphericus and the lateral posterior hypothalamic nucleus demonstrate a pathway between these two structures through the olfactostriatum. Injections in the olfactostriatum and in the medial amygdala show parallel projections to the lateral posterior hypothalamic nucleus. Since this hypothalamic nucleus has been previously described as projecting to the hypoglossal nucleus, both, the medial amygdala and the

  5. Evolution of spatially coexpressed families of type-2 vomeronasal receptors in rodents.

    Science.gov (United States)

    Francia, Simona; Silvotti, Lucia; Ghirardi, Filippo; Catzeflis, François; Percudani, Riccardo; Tirindelli, Roberto

    2014-12-23

    The vomeronasal organ (VNO) is an olfactory structure for the detection of pheromones. VNO neurons express three groups of unrelated G-protein-coupled receptors. Type-2 vomeronasal receptors (V2Rs) are specifically localized in the basal neurons of the VNO and are believed to sense protein pheromones eliciting specific reproductive behaviors. In murine species, V2Rs are organized into four families. Family-ABD V2Rs are expressed monogenically and coexpress with family-C V2Rs of either subfamily C1 (V2RC1) or subfamily C2 (V2RC2), according to a coordinate temporal diagram. Neurons expressing the phylogenetically ancient V2RC1 coexpress family-BD V2Rs or a specific group of subfamily-A V2Rs (V2RA8-10), whereas a second neuronal subset (V2RC2-positive) coexpresses a recently expanded group of five subfamily-A V2Rs (V2RA1-5) along with vomeronasal-specific Major Histocompatibility Complex molecules (H2-Mv). Through database mining and Sanger sequencing, we have analyzed the onset, diversification, and expansion of the V2R-families throughout the phylogeny of Rodentia. Our results suggest that the separation of V2RC1 and V2RC2 occurred in a Cricetidae ancestor in coincidence with the evolution of the H2-Mv genes; this phylogenetic event did not correspond with the origin of the coexpressing V2RA1-5 genes, which dates back to an ancestral myomorphan lineage. Interestingly, the evolution of receptors within the V2RA1-5 group may be implicated in the origin and diversification of some of the V2R putative cognate ligands, the exocrine secreting peptides. The establishment of V2RC2, which probably reflects the complex expansion and diversification of family-A V2Rs, generated receptors that have probably acquired a more subtle functional specificity.

  6. Órgano vomeronasal: Estudio anatómico de prevalencia y su función

    OpenAIRE

    Sarría-Echegaray,Pedro L; Artigas-Sapiaín,Christian E; Rama-López,Julio; soler-Vilarrasa,Ramona; Tomás-Barberán,Manuel D

    2014-01-01

    Introducción: El órgano vomeronasal (OVN) descrito por Jacobson en mamíferos distintos al ser humano, es una incógnita tanto en lo que se refiere a su localización así como a su función en la raza humana. Se considera como un vestigio del olfato, que en los animales mamíferos parece influir en los hábitos sexuales (feromonas) y sociales. Hasta la fecha han sido escasos los estudios concluyentes al respecto en humanos. Objetivo: Conocer la prevalencia del órgano vomeronasal en nuestras consult...

  7. Contrasted Evolution of the Vomeronasal Receptor Repertoires in Mammals and Squamate Reptiles

    OpenAIRE

    Brykczynska, Urszula; Tzika, Athanasia C; Rodriguez, Ivan; Michel C Milinkovitch

    2013-01-01

    The vomeronasal organ (VNO) is an olfactory structure that detects pheromones and environmental cues. It consists of sensory neurons that express evolutionary unrelated groups of transmembrane chemoreceptors. The predominant V1R and V2R receptor repertoires are believed to detect airborne and water-soluble molecules, respectively. It has been suggested that the shift in habitat of early tetrapods from water to land is reflected by an increase in the ratio of V1R/V2R genes. Snakes, which have ...

  8. Human pheromone detection by the vomeronasal organ: unnecessary for mate selection?

    Science.gov (United States)

    Mast, Thomas G; Samuelsen, Chad L

    2009-07-01

    Recently, Foltan and Sedy proposed a hypothesis stating that the adult human VNO is integral to the prevention of inappropriate mate selection. In this commentary, we address the authors' assumption that humans have a functional VNO, that pheromones are detected exclusively by the VNO, and that human pheromones are responsible for negative stimuli during mate selection. After examining the published literature on human vomeronasal function, we argue that their hypothesis is critically flawed. We offer a brief review of the adult human VNO in support of our argument.

  9. The "olfactostriatum" of snakes: a basal ganglia vomeronasal structure in tetrapods.

    Science.gov (United States)

    Martinez-Marcos, Alino; Ubeda-Bañon, Isabel; Lanuza, Enrique; Halpern, Mimi

    2005-09-15

    The olfactostriatum is a portion of the basal ganglia of snakes situated ventromedially to the nucleus accumbens proper. It receives a major vomeronasal input from the nucleus sphericus, the primary target of accessory olfactory bulb efferents. Recently, the ophidian olfactostriatum has been characterized on the basis of chemoarchitecture (distribution of serotonin, neuropeptide Y and tyrosine hydroxylase) and hodology (afferent and efferent connections). In contrast to the nucleus accumbens proper, the olfactostriatum is densely immunoreactive for serotonin and neuropeptide Y and sparsely immunoreactive for tyrosine hydroxylase. The nucleus accumbens proper and the olfactostriatum share most afferent connections except those originating in the nucleus sphericus, which are exclusively directed to the olfactostriatum. Similarly, the nucleus accumbens proper and the olfactostriatum show a similar pattern of efferent connections including those going to the ventral pallidum, although the olfactostriatum alone projects to the main and accessory olfactory bulbs as well as some amygdaloid nuclei. On the basis of its chemoarchitecture, the olfactostriatum resembles the mammalian ventral pallidum (but also the shell of the nucleus accumbens). Its connections, however, suggests that the olfactostriatum could be a specialized portion of the shell of nucleus accumbens extended more ventromedially than previously believed and devoted to processing vomeronasal information. Comparative data suggest that a similar structure is present in the basal ganglia of amphibians and mammals.

  10. The risk of extrapolation in neuroanatomy: the case of the mammalian vomeronasal system

    Directory of Open Access Journals (Sweden)

    Ignacio Salazar

    2009-10-01

    Full Text Available The sense of smell plays a crucial role in mammalian social and sexual behaviour, identification of food, and detection of predators. Nevertheless, mammals vary in their olfactory ability. One reason for this concerns the degree of development of their pars basalis rhinencephali, an anatomical feature that has has been considered in classifying this group of animals as macrosmatic, microsmatic or anosmatic. In mammals, different structures are involved in detecting odours: the main olfactory system, the vomeronasal system (VNS, and two subsystems, namely the ganglion of Grüneberg and the septal organ. Here, we review and summarise some aspects of the comparative anatomy of the VNS and its putative relationship to other olfactory structures. Even in the macrosmatic group, morphological diversity is an important characteristic of the VNS, specifically of the vomeronasal organ and the accessory olfactory bulb. We conclude that it is a big mistake to extrapolate anatomical data of the VNS from species to species, even in the case of relatively close evolutionary proximity between them. We propose to study other mammalian VNS than those of rodents in depth as a way to clarify its exact role in olfaction. Our experience in this field leads us to hypothesise that the VNS, considered for all mammalian species, could be a system undergoing involution or regression, and could serve as one more integrated olfactory subsystem.

  11. Developmental changes affecting lectin binding in the vomeronasal organ of domestic pigs, Sus scrofa.

    Science.gov (United States)

    Park, Junwoo; Lee, Wonho; Jeong, Chanwoo; Kim, Hwangryong; Taniguchi, Kazumi; Shin, Taekyun

    2012-01-01

    This study investigated the developmental changes of glycoconjugate patterns in the porcine vomeronasal organs (VNOs) and associated glands (Jacobson's glands) from prenatal (9 weeks of gestation) and postnatal (2 days after birth) to the sexually mature stage (6 months old). The VNO of pigs (Sus scrofa) was examined using the following: Dolichos biflorus agglutinin (DBA), Bandeiraea simplicifolia agglutinin isolectin B4 (BSI-B4), Triticum vulgaris agglutinin (WGA), Ulex europaeus agglutinin I (UEA-I), and soybean agglutinin (SBA). At the fetal stage, all lectins examined were detected mainly in the free border of the vomeronasal epithelium, but few (WGA and UEA-I) and or absent in the VNO cell bodies. At the postnatal and sexually mature stages, the reactivity of some lectins, including WGA, UEA-I, DBA and SBA, were shown to increase in the VNO sensory epithelium as well as the free border. The increased reactivity of lectins as development progressed was also observed in Jacobson's gland acini. These findings suggest that binding sites of lectins, including those of WGA, UEA-I, DBA, and SBA, increase during development from fetal to postnatal growth, possibly contributing to the increased ability of chemoreception in the pig.

  12. MRI of intact plants.

    NARCIS (Netherlands)

    As, H. van; Scheenen, T.W.J.; Vergeldt, F.J.

    2009-01-01

    Nuclear magnetic resonance imaging (MRI) is a non-destructive and non-invasive technique that can be used to acquire two- or even three-dimensional images of intact plants. The information within the images can be manipulated and used to study the dynamics of plant water relations and water transpor

  13. MRI of intact plants

    NARCIS (Netherlands)

    As, van H.; Scheenen, T.; Vergeldt, F.J.

    2009-01-01

    Nuclear magnetic resonance imaging (MRI) is a non-destructive and non-invasive technique that can be used to acquire two- or even three-dimensional images of intact plants. The information within the images can be manipulated and used to study the dynamics of plant water relations and water transpor

  14. [Intact cervical pregnancy].

    Science.gov (United States)

    Habek, D; Bobic, M V; Dosen, L

    2003-01-01

    The authors describe a case of intact cervical pregnancy in a 24-year-old secundigravida. The patient was treated successfully with Methotrexate. Conservative treatment is the first choice in the therapy of uncomplicated cervical pregnancy. Conservative and operative therapeutic procedures are discussed.

  15. Dog and mouse: Towards a balanced view of the mammalian olfactory system

    Directory of Open Access Journals (Sweden)

    William Arthur Barrios Santos

    2014-09-01

    Full Text Available Although the most intensively studied mammalian olfactory system is that of the mouse, in which olfactory chemical cues of one kind or another are detected in four different nasal areas (the main olfactory epithelium, the septal organ, Grüneberg’s ganglion, and the sensory epithelium of the vomeronasal organ, the extraordinarily sensitive olfactory system of the dog is also an important model that is increasingly used, for example in genomic studies of species evolution. Here we describe the topography and extent of the main olfactory and vomeronasal sensory epithelia of the dog, and we report finding no structures equivalent to the Grüneberg ganglion and septal organ of the mouse. Since we examined adults, newborns and foetuses we conclude that these latter structures are absent in dogs, possibly as the result of regression or involution.The absence of a vomeronasal component based on VR2 receptors suggests that the vomeronasal organ may be undergoing a similar involutionary process.

  16. Identification of G protein α subunits in the main olfactory system and vomeronasal system of the Japanese Striped snake, Elaphe quadrivirgata.

    Science.gov (United States)

    Kondoh, Daisuke; Koshi, Katsuo; Ono, Hisaya K; Sasaki, Kuniaki; Nakamuta, Nobuaki; Taniguchi, Kazuyuki

    2013-01-01

    In the olfactory system, G proteins couple to the olfactory receptors, and G proteins expressed in the main olfactory system and vomeronasal system vary according to animal species. In this study, G protein α subunits expressed in the main olfactory system and vomeronasal system of the snake were identified by immunohistochemistry. In the olfactory epithelium, only anti-Gαolf/s antibody labeled the cilia of the receptor cells. In the vomeronasal epithelium, only anti-Gαo antibody labeled the microvilli of the receptor cells. In the accessory olfactory bulb, anti-Gαo antibody stained the whole glomerular layer. These results suggest that the main olfactory system and the vomeronasal system of the snake express Gαolf and Gαo as G proteins coupling to the olfactory receptors, respectively.

  17. [Late embryonic development of the vomeronasal complex of the cat (Felis silvestris)].

    Science.gov (United States)

    Wöhrmann-Repenning, A; Ciba, B

    1989-01-01

    An examination of 2 feline embryos in different stages of development (overall length 60 and 115 mm respectively) reveals a well developed vomeronasal complex in each case. Jacobson's Organs embedded within the paraseptal cartilage form long blind tubes at the base of the septum nasi. The cartilage is caudally tub-shaped and embraces rostrally completely the organ over a considerable length. In this manner a long, nearly tunnel-like tube is formed which represents a modified form of the original outer bar and which has not been described so far in cats. It stretches rostro-ventrally across the branching region of the paraseptal cartilage as far as the mouth of Jacobson's Organ. The dorsal branch of the cartilago paraseptalis on the other hand forms a vertically oriented strip which connects to the lamina transversalis anterior. The ductus nasopalatinus passing through the palate is laterally supported by a cartilago ductus nasopalatini which rostrally to the mouth of Jacobson's Organ forms a unified element with the ventral branch of the cartilago paraseptalis. In the case of the younger cat embryo, this cartilago ductus nasopalatini is yet weakly developed. The ductus nasopalatini of the embryos studied are in an amazingly retarded state of development. The ductus, which are blocked in the early stages of the embryonic development during secondary palate formation, form predominantly solid strands of epithelium. By dissolving the cemented epithelium, the ductus are open. But even in the case of the older embryo of the cat, this process is not completed yet. The short duct connecting Jacobson's Organ with the ductus nasopalatinus is also still closed in both embryos. Such cemented sections of epithelium of the younger embryo reveals an interesting relation between the ductus nasopalatinus and the ductus nasolacrimalis which so far has not been pointed out for mammals. From the point of view of phylogenetics, the locally specialized vomeronasal complex of cats

  18. (Photosynthesis in intact plants)

    Energy Technology Data Exchange (ETDEWEB)

    1990-01-01

    Progress in the two years since the last renewal application has been excellent. We have made substantial contributions on both main fronts of the projects, and are particularly happy with the progress of our research on intact plants. The approach of basing our field work on a sound foundation of laboratory studies has enabled is to use methods which provide unambiguous assays of well characterized reactions. We have also made excellent progress in several laboratory studies which will have direct applications in future field work, and have introduced to the laboratory a range of molecular genetics techniques which will allow us to explore new options in the attempt to understand function at the level of molecular structure.

  19. Origin of the Genetic Components of the Vomeronasal System in the Common Ancestor of all Extant Vertebrates

    Science.gov (United States)

    Grus, Wendy E.; Zhang, Jianzhi

    2009-01-01

    Comparative genomics provides a valuable tool for inferring the evolutionary history of physiological systems, particularly when this information is difficult to ascertain by morphological traits. One such example is the vomeronasal system (VNS), a vertebrate nasal chemosensory system that is responsible for detecting intraspecific pheromonal cues as well as environmental odorants. The morphological components of the VNS are found only in tetrapods, but the genetic components of the system have been found in teleost fish, in addition to tetrapods. To determine when the genetic components of the VNS originated, we searched for the VNS-specific genes in the genomes of two early diverging vertebrate lineages: the sea lamprey from jawless fishes and the elephant shark from cartilaginous fishes. Genes encoding vomeronasal type 1 receptors (V1Rs) and Trpc2, two components of the vomeronasal signaling pathway, are present in the sea lamprey genome, and both are expressed in the olfactory organ, revealing that the genetic components of the present-day VNS existed in the common ancestor of all extant vertebrates. Additionally, all three VNS genes, Trpc2, V1Rs, and vomeronasal type 2 receptors (V2Rs), are found in the elephant shark genome. Because V1Rs and V2Rs are related to two families of taste receptors, we also searched the early diverging vertebrate genomes for taste system genes and found them in the shark genome but not in the lamprey. Coupled with known distributions of the genetic components of the vertebrate main olfactory system, our results suggest staggered origins of vertebrate sensory systems. These findings are important for understanding the evolution of vertebrate sensory systems and illustrate the utility of the genome sequences of early diverging vertebrates for uncovering the evolution of vertebrate-specific traits. PMID:19008528

  20. El sistema vomeronasal y su posible funcionalidad en larvas de anuros

    Directory of Open Access Journals (Sweden)

    Pozzi, Andrea Gabriela

    2012-12-01

    Full Text Available El sistema vomeronasal (SVN es un sistema olfatorio accesorio presente en la mayoría de los tetrápodos. Clásicamente se lo ha asociado con el paso de los vertebrados al ambiente terrestre; sin embargo las evidencias surgidas en los últimos años indican que el SVN apareció tempranamente en la evolución de los tetrápodos y sería funcional en ambientes acuáticos. Este sistema sensorial ha sido descripto en etapas larvales de anuros. Pero ¿es funcional el SVN en renacuajos? No existen experimentos en donde se evalúe la participación del SVN en la quimiodetección en renacuajos. Sin embargo, un número considerable de evidencias indican que este sistema sensorial podría ser funcional en larvas de anuros: 1 El órgano vomeronasal (OVN aparece durante el desarrollo embrionario y está presente durante toda la etapa larval. 2 El OVN contiene neuronas bipolares cuyos axones proyectan al bulbo olfatorio accesorio (BOA donde establecen conexiones sinápticas con neuronas telencefálicas. 3 Las neuronas del OVN expresan los receptores de membrana descriptos en tetrápodos, así como la proteína G involucrada en la señalización intracelular. 4 Los análisis de microscopía electrónica demuestran que el OVN posee neuronas con microvellosidades apicales como se describe para otros grupos y sus características ultraestructurales no se modifican durante la metamorfosis. Más aún, no hay diferencias en la ultraestructura de las conexiones sinápticas entre larvas y adultos a nivel del BOA. Los renacuajos presentan una gran cantidad de comportamientos mediados por quimiodetección. Conocer si el SVN participa en la detección de alguno/s de estos estímulos ayudaría no sólo a dilucidar aspectos relacionados con la comunicación química y el comportamiento en renacuajos sino también a comprender aspectos evolutivos de los sistemas quimiosensoriales en vertebrados.

  1. Afferent and efferent connections of the nucleus sphericus in the snake Thamnophis sirtalis: convergence of olfactory and vomeronasal information in the lateral cortex and the amygdala.

    Science.gov (United States)

    Lanuza, E; Halpern, M

    1997-09-08

    This paper is an account of the afferent and efferent projections of the nucleus sphericus (NS), which is the major secondary vomeronasal structure in the brain of the snake Thamnophis sirtalis. There are four major efferent pathways from the NS: 1) a bilateral projection that courses, surrounding the accessory olfactory tract, and innervates several amygdaloid nuclei (nucleus of the accessory olfactory tract, dorsolateral amygdala, external amygdala, and ventral anterior amygdala), the rostral parts of the dorsal and lateral cortices, and the accessory olfactory bulb; 2) a bilateral projection that courses through the medial forebrain bundle and innervates the olfactostriatum (rostral and ventral striatum); 3) a commissural projection that courses through the anterior commissure and innervates mainly the contralateral NS; and 4) a meager bilateral projection to the lateral hypothalamus. On the other hand, important afferent projections to the NS arise solely in the accessory olfactory bulb, the nucleus of the accessory olfactory tract, and the contralateral NS. This pattern of connections has three important implications: first, the lateral cortex probably integrates olfactory and vomeronasal information. Second, because the NS projection to the hypothalamus is meager and does not reach the ventromedial hypothalamic nucleus, vomeronasal information from the NS is not relayed directly to that nucleus, as previously reported. Finally, a structure located in the rostral and ventral telencephalon, the olfactostriatum, stands as the major tertiary vomeronasal center in the snake brain. These three conclusions change to an important extent our previous picture of how vomeronasal information is processed in the brain of reptiles.

  2. Voltage-dependent currents in microvillar receptor cells of the frog vomeronasal organ.

    Science.gov (United States)

    Trotier, D; Døving, K B; Rosin, J F

    1993-08-01

    Vomeronasal receptor cells are differentiated bipolar neurons with a long dendrite bearing numerous microvilli. Isolated cells (with a mean dendritic length of 65 microns) and cells in mucosal slices were studied using whole-cell and Nystatin-perforated patch-clamp recordings. At rest, the membrane potential was -61 +/- 13 mV (mean +/- SD; n = 61). Sixty-four per cent of the cells had a resting potential in the range of -60 to -86 mV, with almost no spontaneous action potential. The input resistance was in the G omega range and overshooting repetitive action potentials were elicited by injecting depolarizing current pulses in the range of 2-10 pA. Voltage-dependent currents were characterized under voltage-clamp conditions. A transient fast inward current activating near -45 mV was blocked by tetrodotoxin. In isolated cells, it was half-deactivated at a membrane potential near -75 mV. An outward K+ current was blocked by internal Cs+ ions or by external tetraethylammonium or Ba2+ ions. A calcium-activated voltage-dependent potassium current was blocked by external Cd2+ ions. A voltage-dependent Ca2+ current was observed in an iso-osmotic BaCl2 solution. Finally, a hyperpolarization-activated inward current was recorded. Voltage-dependent currents in these microvillar olfactory receptor neurons appear qualitatively similar to those already described in ciliated olfactory receptor cells located in the principal olfactory epithelium.

  3. A candidate subspecies discrimination system involving a vomeronasal receptor gene with different alleles fixed in M. m. domesticus and M. m. musculus.

    Directory of Open Access Journals (Sweden)

    Robert C Karn

    Full Text Available Assortative mating, a potentially efficient prezygotic reproductive barrier, may prevent loss of genetic potential by avoiding the production of unfit hybrids (i.e., because of hybrid infertility or hybrid breakdown that occur at regions of secondary contact between incipient species. In the case of the mouse hybrid zone, where two subspecies of Mus musculus (M. m. domesticus and M. m. musculus meet and exchange genes to a limited extent, assortative mating requires a means of subspecies recognition. We based the work reported here on the hypothesis that, if there is a pheromone sufficiently diverged between M. m. domesticus and M. m. musculus to mediate subspecies recognition, then that process must also require a specific receptor(s, also sufficiently diverged between the subspecies, to receive the signal and elicit an assortative mating response. We studied the mouse V1R genes, which encode a large family of receptors in the vomeronasal organ (VNO, by screening Perlegen SNP data and identified one, Vmn1r67, with 24 fixed SNP differences most of which (15/24 are nonsynonymous nucleotide substitutions between M. m. domesticus and M. m. musculus. We observed substantial linkage disequilibrium (LD between Vmn1r67 and Abpa27, a mouse salivary androgen-binding protein gene that encodes a proteinaceous pheromone (ABP capable of mediating assortative mating, perhaps in conjunction with its bound small lipophilic ligand. The LD we observed is likely a case of association rather than residual physical linkage from a very recent selective sweep, because an intervening gene, Vmn1r71, shows significant intra(subspecific polymorphism but no inter(subspecific divergence in its nucleotide sequence. We discuss alternative explanations of these observations, for example that Abpa27 and Vmn1r67 are coevolving as signal and receptor to reinforce subspecies hybridization barriers or that the unusually divergent Vmn1r67 allele was not a product of fast positive

  4. Serum steroid levels in intact and endocrine ablated BALB/c nude mice and their intact littermates

    DEFF Research Database (Denmark)

    Brünner, N; Svenstrup, B; Spang-Thomsen, M;

    1986-01-01

    An investigation was made of the serum steroid levels found in intact and endocrine ablated nude mice of both sexes and in their intact homozygous littermates. The results showed that nude mice have a normal steroidogenesis, but with decreased levels of circulating steroids compared to those...... of the littermates. The efficacy of the endocrine ablations was confirmed by the reduction in serum oestrone following oophorectomy, and by the reduction in serum testosterone and progesterone following orchiectomy. The normal steroidogenesis in nude mice, and the similarities between mouse and man with regard...

  5. Chemoarchitecture and afferent connections of the "olfactostriatum": a specialized vomeronasal structure within the basal ganglia of snakes.

    Science.gov (United States)

    Martinez-Marcos, Alino; Ubeda-Bañon, Isabel; Lanuza, Enrique; Halpern, Mimi

    2005-01-01

    The olfactostriatum, a portion of the striatal complex of snakes, is the major tertiary vomeronasal structure in the ophidian brain, receiving substantial afferents from the nucleus sphericus, the primary target of accessory olfactory bulb efferents. In the present study, we have characterized the olfactostriatum of garter snakes (Thamnophis sirtalis) on the basis of chemoarchitecture (distribution of serotonin, neuropeptide Y and tyrosine hydroxylase) and hodology (afferent connections). The olfactostriatum is densely immunoreactive for serotonin and neuropeptide Y and shows moderate-to-weak immunoreactivity for tyrosine hydroxylase. In addition to afferents from the nucleus sphericus, the olfactostriatum receives inputs from the dorsal and lateral cortices, nucleus of the accessory olfactory tract, external and dorsolateral amygdalae, dorsomedial thalamic nucleus, ventral tegmental area and raphe nuclei. Double labeling experiments demonstrated that the distribution of serotonin and neuropeptide Y in this area almost completely overlaps the terminal field of projections from the nucleus sphericus. Also, serotonergic and dopaminergic innervation of the olfactostriatum likely arise, respectively, from the raphe nuclei and the ventral tegmental area, whereas local circuit neurons originate the neuropeptide Y immunoreactivity. These results indicate that the olfactostriatum of snakes could be a portion of the nucleus accumbens, with features characteristic of the accumbens shell, devoted to processing vomeronasal information. Comparative data suggest that a similar structure is present in the ventral striatum of amphibians and mammals.

  6. Sexual polymorphisms of vomeronasal 1 receptor family gene expression in bulls, steers, and estrous and early luteal-phase heifers.

    Science.gov (United States)

    Kubo, Haruna; Otsuka, Midori; Kadokawa, Hiroya

    2016-02-01

    Vomeronasal 1 receptors (V1R) are a family of receptors for intraspecies chemosignals, including pheromones, and are expressed in the olfactory epithelium (OE) and vomeronasal organ (VO). Even in the well-studied rodents, it is unclear which members of the V1R family cause sexual polymorphisms, as there are numerous genes and it is difficult to quantify their expressions individually. Bovine species carry only 34 V1R homologs, and the OE and VOs are large enough to sample. Here, V1R expression was quantified in the OE and VOs of individual bovines. Based on the 34 gene sequences, we obtained a molecular dendrogram consisting of four clusters and six independent branches. Semi-quantitative RT-PCR was used to obtain gene expression profiles in the VOs and OE of 5 Japanese Black bulls, 5 steers, 7 estrous heifers and 6 early luteal-phase heifers. Ten genes showed significant between-group differences, and 22 showed high expression in VOs than in OE. The bulls showed higher expression of one gene more in OE and another in VOs (both Pexpressed more abundantly in steers than in bulls. The estrous heifers showed higher expression of a gene of the second cluster in OE, and a gene of the third cluster in VOs (both Pexpression exhibits sexual polymorphisms in cattle.

  7. Functions of Vomeronasal System in Reproduction Behavior of the Rodent%犁鼻系统在鼠类繁殖行为中的功能

    Institute of Scientific and Technical Information of China (English)

    邰发道; 王廷正; 孙儒泳

    2002-01-01

    通过对犁鼻器在鼠类繁殖行为中的功能研究,并将其与主要嗅觉系统的功能进行比较,发现虽然犁鼻器是一个微小的、而且常常是被人们所忽视并有争论的器官,但它在鼠类行为,尤其在繁殖行为方面具有多种功能.对此要用新的研究方法对犁鼻系统的功能作进一步研究,以便和主要嗅觉系统在社会行为中的作用进行比较,进而更精细地探讨人类犁鼻系统的生理机能.%Functions in rodent reproduction behavior were discussed, difference between functions of vomeronasal system (VNS) and main olfactory system (MOS) in rodent behaviors were compared in this paper. It is found that although vomeronasal organ is a minute structure and often is disregarded and argued by people, it has a wide variety of functions in rodent behavior especially in reproduction behavior. In order to compare the functions of main olfactory system of mammals and accessory olfactory system and to probe physiologic function of vomeronasal system of human beings in detail, it is necessary to study vomeronasal system further using new methods.

  8. Morphological, histochemical and computed tomography on the vomeronasal organ (Jacobson’s organ of Egyptian native breeds of goats (Capra hircus

    Directory of Open Access Journals (Sweden)

    Usama Kamal Moawad

    2017-06-01

    Full Text Available Background: The vomeronasal organ (Jacobson’s organ is a chemosensory paired tubular organ located on both sides of the nasal septum at its base. It plays an essential role in reproduction process and social behaviors. Objective: The current study investigated both anatomical and histological structure of the vomeronasal organ (VNO in Egyptian native breeds of goats using cross sectional anatomy, histological techniques and computed tomography (CT. Methodology: A total of thirty heads obtained from adult and apparently healthy goats of both sexes were collected from Beni-Suef slaughterhouse in Beni-Suef province, Egypt then subjected to anatomical, histological and computed topographical studies. Results: Grossly, the VNO appeared as two blind ducts on the either sides of the nasal septum at the floor of nasal cavity extending from the nasal opening of incisive papilla rostrally to the upper 2nd premolar teeth caudally. It connected with mouth by two nasopalatine ducts. The histological examinations revealed two types of lining epithelium; non sensory type lining the cranial portion and the lateral wall of the middle portion of the vomeronasal duct (VND, whereas an olfactory type was the lining epithelium of the medial wall of the middle and the whole caudal portions. The lamina propria submucosa exhibited vascular loose connective tissues, serous glands, nerve bundles and encapsulated by hyaline cartilage. Conclusion: The obtaining olfactory epithelium in VNO may indicate an essential role of this organ in sexual relationships and sociosexual behaviors through perception of pheromones.

  9. Both olfactory epithelial and vomeronasal inputs are essential for activation of the medial amygdala and preoptic neurons of male rats.

    Science.gov (United States)

    Dhungel, S; Masaoka, M; Rai, D; Kondo, Y; Sakuma, Y

    2011-12-29

    Chemosensory inputs signaling volatile and nonvolatile molecules play a pivotal role in sexual and social behavior in rodents. We have demonstrated that olfactory preference in male rats, that is, attraction to receptive female odors, is regulated by the medial amygdala (MeA), the cortical amygdala (CoA), and the preoptic area (POA). In this paper, we investigated the involvement of two chemosensory organs, the olfactory epithelium (OE) and the vomeronasal organ (VNO), in olfactory preference and copulatory behavior in male rats. We found that olfactory preferences were impaired by zinc sulfate lesion of the OE but not surgical removal of the VNO. Copulatory behaviors, especially intromission frequency and ejaculation, were also suppressed by zinc sulfate treatment. Neuronal activation in the accessory olfactory bulb (AOB), the MeA, the CoA, and the POA was analyzed after stimulation by airborne odors or soiled bedding of estrous females using cFos immunohistochemistry. Although the OE and VNO belong to different neural systems, the main and accessory olfactory systems, respectively, both OE lesion and VNO removal almost equally suppressed the number of cFos-immunoreactive cells in those areas that regulate olfactory preference. These results suggest that signals received by the OE and VNO interact and converge in the early stage of olfactory processing, in the AOB and its targets, although they have distinct roles in the regulation of social behaviors.

  10. Intact Transition Epitope Mapping (ITEM)

    Science.gov (United States)

    Yefremova, Yelena; Opuni, Kwabena F. M.; Danquah, Bright D.; Thiesen, Hans-Juergen; Glocker, Michael O.

    2017-08-01

    Intact transition epitope mapping (ITEM) enables rapid and accurate determination of protein antigen-derived epitopes by either epitope extraction or epitope excision. Upon formation of the antigen peptide-containing immune complex in solution, the entire mixture is electrosprayed to translate all constituents as protonated ions into the gas phase. There, ions from antibody-peptide complexes are separated from unbound peptide ions according to their masses, charges, and shapes either by ion mobility drift or by quadrupole ion filtering. Subsequently, immune complexes are dissociated by collision induced fragmentation and the ion signals of the "complex-released peptides," which in effect are the epitope peptides, are recorded in the time-of-flight analyzer of the mass spectrometer. Mixing of an antibody solution with a solution in which antigens or antigen-derived peptides are dissolved is, together with antigen proteolysis, the only required in-solution handling step. Simplicity of sample handling and speed of analysis together with very low sample consumption makes ITEM faster and easier to perform than other experimental epitope mapping methods.

  11. Bacterial delivery of large intact genomic-DNA-containing BACs into mammalian cells.

    Science.gov (United States)

    Cheung, Wing; Kotzamanis, George; Abdulrazzak, Hassan; Goussard, Sylvie; Kaname, Tadashi; Kotsinas, Athanassios; Gorgoulis, Vassilis G; Grillot-Courvalin, Catherine; Huxley, Clare

    2012-01-01

    Efficient delivery of large intact vectors into mammalian cells remains problematical. Here we evaluate delivery by bacterial invasion of two large BACs of more than 150 kb in size into various cells. First, we determined the effect of several drugs on bacterial delivery of a small plasmid into different cell lines. Most drugs tested resulted in a marginal increase of the overall efficiency of delivery in only some cell lines, except the lysosomotropic drug chloroquine, which was found to increase the efficiency of delivery by 6-fold in B16F10 cells. Bacterial invasion was found to be significantly advantageous compared with lipofection in delivering large intact BACs into mouse cells, resulting in 100% of clones containing intact DNA. Furthermore, evaluation of expression of the human hypoxanthine phosphoribosyltransferase (HPRT) gene from its genomic locus, which was present in one of the BACs, showed that single copy integrations of the HPRT-containing BAC had occurred in mouse B16F10 cells and that expression of HPRT from each human copy was 0.33 times as much as from each endogenous mouse copy. These data provide new evidence that bacterial delivery is a convenient and efficient method to transfer large intact therapeutic genes into mammalian cells.

  12. Bacterial delivery of large intact genomic-DNA-containing BACs into mammalian cells

    Science.gov (United States)

    Cheung, Wing; Kotzamanis, George; Abdulrazzak, Hassan; Goussard, Sylvie; Kaname, Tadashi; Kotsinas, Athanassios; Gorgoulis, Vassilis G.; Grillot-Courvalin, Catherine; Huxley, Clare

    2012-01-01

    Efficient delivery of large intact vectors into mammalian cells remains problematical. Here we evaluate delivery by bacterial invasion of two large BACs of more than 150 kb in size into various cells. First, we determined the effect of several drugs on bacterial delivery of a small plasmid into different cell lines. Most drugs tested resulted in a marginal increase of the overall efficiency of delivery in only some cell lines, except the lysosomotropic drug chloroquine, which was found to increase the efficiency of delivery by 6-fold in B16F10 cells. Bacterial invasion was found to be significantly advantageous compared with lipofection in delivering large intact BACs into mouse cells, resulting in 100% of clones containing intact DNA. Furthermore, evaluation of expression of the human hypoxanthine phosphoribosyltransferase (HPRT) gene from its genomic locus, which was present in one of the BACs, showed that single copy integrations of the HPRT-containing BAC had occurred in mouse B16F10 cells and that expression of HPRT from each human copy was 0.33 times as much as from each endogenous mouse copy. These data provide new evidence that bacterial delivery is a convenient and efficient method to transfer large intact therapeutic genes into mammalian cells. PMID:22095052

  13. Intacting Integrity in coping with health issues

    DEFF Research Database (Denmark)

    Jepsen, Stine Leegaard; Bastrup Jørgensen, Lene; Fridlund, Bengt

    2017-01-01

    The aim of this study was to develop a formal substantive theory (FST) on the multidimensional behavioral process of coping with health issues. Intacting integrity while coping with health issues emerged as the core category of this FST. People facing health issues strive to safeguard and keep...... intact their integrity not only on an individual level but also as members of a group or a system. This intacting process is executed by attunement, continuously minimizing the discrepancy between personal values, personal health, self-expectations and external conditions as health- and cultural...

  14. NCBI nr-aa BLAST: CBRC-MMUS-06-0126 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0126 sp|Q8VBS7|VN1A6_MOUSE Vomeronasal type-1 receptor A6 (Vomeronasal... type-1 receptor A3) (Vomeronasal type-1 receptor A1) (Vomeronasal type-1 receptor A10) dbj|BAB79209.1| vomeronasal... receptor 1 A1 [Mus musculus] dbj|BAB79210.1| vomeronasal receptor 1 A3 [Mus musculus] gb|AAK98772.1| vomeronasal

  15. 菜花原矛头蝮嗅觉系统和犁鼻系统的显微结构%The microstructure of olfactory and vomeronasal systems in snake, Protobothrops jerdonii

    Institute of Scientific and Technical Information of China (English)

    王宏元; 柴丽红; 王晓雯; 李忻怡

    2011-01-01

    采用组织学方法观察了菜花原矛头蝮(Protobothrops jerdonii)的嗅觉系统和犁鼻系统.结果表明,嗅器位于嗅囊的背侧,犁鼻器则位于嗅器的腹内侧.嗅器的上皮分化为嗅上皮和呼吸上皮,嗅上皮的基底层有Bowman's腺,呼吸上皮中具有大量的杯状细胞.犁鼻器基底层未发现犁鼻腺.嗅球和副嗅球呈典型的板层构筑结构.推测菜花原矛头蝮嗅器内嗅上皮和呼吸上皮完全分开有利于背侧嗅上皮俘获气味信号,腹侧呼吸上皮参与呼吸作用.虽然菜花原矛头蝮等蛇类的犁鼻器缺少犁鼻腺,但是其眼眶周围的哈氏腺和口腔内的唾液腺可以代偿犁鼻腺机能.%The olfactory organ and vomeronasal system in snake, Protobothrops jerdonii, were investigated under the microscope. The results showed that the olfactory organ lies in the dorsal and the vomeronasal organ is in the ventral portion of the nose cavity. The epithelium of the olfactory cavity is divided into the two segments: olfactory epithelium and respiratory epithelium.Bowman's gland in the olfactory epithelium is present and goblets cells occur in the respiratory epithelium. Vomeronasal gland is not found in the vomeronasal organ. The olfactory bulb and accessory olfactory bulb are typical laminar pattern. Subdivision of dorsal olfactory epithelium and ventral respiratory epithelium in snake may be beneficial to that the olfactory epithelium capture odour and respiratory epithelium take party into the respiratory. In addition, the vomeronasal organ in snake lacks vomeronasal glands, but the harderian gland and the salivary glands compensate lack of the vomeronasal gland.

  16. Autism Spectrum Disorder and intact executive functioning.

    Science.gov (United States)

    Ferrara, R; Ansermet, F; Massoni, F; Petrone, L; Onofri, E; Ricci, P; Archer, T; Ricci, S

    2016-01-01

    Earliest notions concerning autism (Autism Spectrum Disorders, ASD) describe the disturbance in executive functioning. Despite altered definition, executive functioning, expressed as higher cognitive skills required complex behaviors linked to the prefrontal cortex, are defective in autism. Specific difficulties in children presenting autism or verbal disabilities at executive functioning levels have been identified. Nevertheless, the developmental deficit of executive functioning in autism is highly diversified with huge individual variation and may even be absent. The aim of the present study to examine the current standing of intact executive functioning intact in ASD.

  17. Xenopus egg cytoplasm with intact actin.

    Science.gov (United States)

    Field, Christine M; Nguyen, Phuong A; Ishihara, Keisuke; Groen, Aaron C; Mitchison, Timothy J

    2014-01-01

    We report optimized methods for preparing Xenopus egg extracts without cytochalasin D, that we term "actin-intact egg extract." These are undiluted egg cytoplasm that contains abundant organelles, and glycogen which supplies energy, and represents the least perturbed cell-free cytoplasm preparation we know of. We used this system to probe cell cycle regulation of actin and myosin-II dynamics (Field et al., 2011), and to reconstitute the large, interphase asters that organize early Xenopus embryos (Mitchison et al., 2012; Wühr, Tan, Parker, Detrich, & Mitchison, 2010). Actin-intact Xenopus egg extracts are useful for analysis of actin dynamics, and interaction of actin with other cytoplasmic systems, in a cell-free system that closely mimics egg physiology, and more generally for probing the biochemistry and biophysics of the egg, zygote, and early embryo. Detailed protocols are provided along with assays used to check cell cycle state and tips for handling and storing undiluted egg extracts.

  18. Myocardial ischemia, reperfusion, and infarction in chronically instrumented, intact, conscious, and unrestrained mice.

    Science.gov (United States)

    Lujan, Heidi L; Janbaih, Hussein; Feng, Han-Zhong; Jin, Jian-Ping; DiCarlo, Stephen E

    2012-06-15

    In the United States alone, the National Heart, Lung, and Blood Institute (NHLBI) has invested several hundred million dollars in pursuit of myocardial infarct-sparing therapies. However, due largely to methodological limitations, this investment has not produced any notable clinical application or cardioprotective therapy. Among the major methodological limitations is the reliance on animal models that do not mimic the clinical situation. In this context, the limited use of conscious animal models is of major concern. In fact, whenever possible, studies of cardiovascular physiology and pathophysiology should be conducted in conscious, complex models to avoid the complications associated with the use of anesthesia and surgical trauma. The mouse has significant advantages over other experimental models for the investigation of infarct-sparing therapies. The mouse is inexpensive, has a high throughput, and presents the ability of one to create genetically modified models. However, successful infarct-sparing therapies in anesthetized mice or isolated mouse hearts may not be successful in more complex models, including conscious mice. Accordingly, a conscious mouse model of myocardial ischemia and reperfusion has the potential to be of major importance for advancing the concepts and methods that drive the development of infarct-sparing therapies. Therefore, we describe, for the first time, the use of an intact, conscious, and unrestrained mouse model of myocardial ischemia-reperfusion and infarction. The conscious mouse model permits occlusion and reperfusion of the left anterior descending coronary artery in an intact, complex model free of the confounding influences of anesthetics and surgical trauma. This methodology may be adopted for advancing the concepts and ideas that drive cardiovascular research.

  19. NCBI nr-aa BLAST: CBRC-MMUS-06-0137 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0137 sp|Q8VBS7|VN1A6_MOUSE Vomeronasal type-1 receptor A6 (Vomeronasal... type-1 receptor A3) (Vomeronasal type-1 receptor A1) (Vomeronasal type-1 receptor A10) dbj|BAB79209.1| vomeronasa...l receptor 1 A1 [Mus musculus] dbj|BAB79210.1| vomeronasal receptor 1 A3 [Mus musculus] gb|AAK98772.1| vomeronasa

  20. How can we conserve intact tropical peatlands?

    Science.gov (United States)

    Lawson, Ian; Roucoux, Katherine

    2017-04-01

    The scientific community has, for more than three decades, been expressing increasing alarm about the fate of peatlands in parts of Indonesia and Malaysia, where extensive land-use conversion and drainage for rice and oil palm have greatly compromised peatland hydrology, ecology, biological richness, and carbon storage. The discourse in the literature on these peatlands is now moving on from attempts to preserve the last remaining fragments of peat-swamp forest, towards discussion of how best to restore damaged ecosystems, and whether it is possible to manage plantations more 'sustainably'. It is becoming increasingly clear, however, that peatlands occur quite widely in other parts of the lowland tropics, including parts of Amazonia and the Congo Basin, and many of these peatlands can reasonably be described as 'intact': although few if any parts of the tropics are totally unaffected by human actions, the hydrology and functional ecology of these systems appear to be close to a 'natural' state. The question then arises as to what should be done with the knowledge of their existence. Here we analyse the arguments in favour of protecting intact peatlands, and the potential conflicts with other priorities such as economic development and social justice. We evaluate alternative mechanisms for protecting intact peatlands, focusing on the particular issues raised by peatlands as opposed to other kinds of tropical ecosystem. We identify ways in which natural science agendas can help to inform these arguments, using our own contributions in palaeoecology and carbon mapping as examples. Finally, we argue for a radical reconsideration of research agendas in tropical peatlands, highlighting the potential contribution of methodologies borrowed from the social sciences and humanities.

  1. Modified alkaline elution allows the measurement of intact apurinic sites in mammalian genomic DNA.

    Science.gov (United States)

    Malvy, C; Lefrançois, M; Bertrand, J R; Markovits, J

    2000-08-01

    The presence of apurinic/apyrimidinic (AP) sites in cell genomes is known to be toxic and mutagenic. These lesions are therefore repaired in cells by efficient enzymatic systems. However, a report (Nakamura and Swenberg, Cancer Res. 59 (1999) 2522-2526) indicates an unexpected high rate of endogenous apurinic/apyrimidinic (AP) sites in genomic DNA in mammalian tissues. The technology used does not allow the authors to distinguish between intact AP sites and 3'cleaved AP sites. The corresponding values range between 2 and 4 sites per million of nucleotides in various human and rat tissues. Using a modified alkaline elution method we show here that the stationary level of intact AP sites is about 0.16 per million of nucleotides in leukemic mouse L1210 cells.

  2. Recollections of Parent Characteristics and Attachment Patterns for College Women of Intact vs. Non-Intact Families

    Science.gov (United States)

    Kilmann, Peter R.; Carranza, Laura V.; Vendemia, Jennifer M. C.

    2006-01-01

    This study contrasted offsprings' attachment patterns and recollections of parent characteristics in two college samples: 147 females from intact biological parents and 157 females of parental divorce. Secure females from intact or non-intact families rated parents positively, while insecure females rated parents as absent, distant, and demanding.…

  3. Robo-2 controls the segregation of a portion of basal vomeronasal sensory neuron axons to the posterior region of the accessory olfactory bulb.

    Science.gov (United States)

    Prince, Janet E A; Cho, Jin Hyung; Dumontier, Emilie; Andrews, William; Cutforth, Tyler; Tessier-Lavigne, Marc; Parnavelas, John; Cloutier, Jean-François

    2009-11-11

    The ability of sensory systems to detect and process information from the environment relies on the elaboration of precise connections between sensory neurons in the periphery and second order neurons in the CNS. In mice, the accessory olfactory system is thought to regulate a wide variety of social and sexual behaviors. The expression of the Slit receptors Robo-1 and Robo-2 in vomeronasal sensory neurons (VSNs) suggests they may direct the stereotypic targeting of their axons to the accessory olfactory bulb (AOB). Here, we have examined the roles of Robo-1 and Robo-2 in the formation of connections by VSN axons within the AOB. While Robo-1 is not necessary for the segregation of VSN axons within the anterior and posterior regions of the AOB, Robo-2 is required for the targeting of some basal VSN axons to the posterior region of the AOB but is dispensable for the fasciculation of VSN axons. Furthermore, the specific ablation of Robo-2 expression in VSNs leads to mistargeting of a portion of basal VSN axons to the anterior region of the AOB, indicating that Robo-2 expression is required on projecting VSN axons. Together, these results identify Robo-2 as a receptor that controls the targeting of basal VSN axons to the posterior AOB.

  4. NIH mouse study finds gut microorganisms may determine cancer treatment outcome

    Science.gov (United States)

    An intact gut commensal microbiota, which is a population of microorganisms living in the intestine, is required for optimal response to cancer therapy, according to a mouse study by scientists at the National Cancer Institute (NCI)

  5. Effects of hydrolysed casein, intact casein and intact whey protein on energy expenditure and appetite regulation

    DEFF Research Database (Denmark)

    Bendtsen, Line Quist; Lorenzen, Janne Kunchel; Gomes, Sisse

    2014-01-01

    Casein and whey differ in amino acid composition and in the rate of absorption; however, the absorption rate of casein can be increased to mimic that of whey by exogenous hydrolysis. The objective of the present study was to compare the effects of hydrolysed casein (HC), intact casein (IC......) and intact whey (IW) on energy expenditure (EE) and appetite regulation, and thereby to investigate the influence of amino acid composition and the rate of absorption. In the present randomised cross-over study, twenty-four overweight and moderately obese young men and women consumed three isoenergetic...... dietary treatments that varied in protein source. The study was conducted in a respiration chamber, where EE, substrate oxidation and subjective appetite were measured over 24 h at three independent visits. Moreover, blood and urine samples were collected from the participants. The results showed...

  6. Comparison of Intact PTH and Bio-Intact PTH Assays Among Non-Dialysis Dependent Chronic Kidney Disease Patients.

    Science.gov (United States)

    Einbinder, Yael; Benchetrit, Sydney; Golan, Eliezer; Zitman-Gal, Tali

    2017-09-01

    The third-generation bio-intact parathyroid hormone (PTH) (1-84) assay was designed to overcome problems associated with the detection of C-terminal fragments by the second-generation intact PTH assay. The two assays have been compared primarily among dialysis populations. The present study evaluated the correlations and differences between these two PTH assays among patients with chronic kidney disease (CKD) stages 3 to 5 not yet on dialysis. Blood samples were collected from 98 patients with CKD stages 3 to 5. PTH concentrations were measured simultaneously by using the second-generation - PTH intact-STAT and third-generation bio-intact 1-84 PTH assays. Other serum biomarkers of bone mineral disorders were also assessed. CKD stage was calculated by using the CKD-Epidemiology Collaboration (EPI) formula. Serum bio-intact PTH concentrations were strongly correlated but significantly lower than the intact PTH concentrations (r=0.963, Pbio-intact PTH) positively correlated with urea (r=0.523, r=0.504; P=0.002, respectively), phosphorus (r=0.532, r=0.521; Pbio-intact PTH assay detected significantly lower PTH concentrations compared with intact PTH assay. Additional studies that correlate the diagnosis and management of CKD mineral and bone disorders with bone histomorphometric findings are needed to determine whether bio-intact PTH assay results are better surrogate markers in these early stages of CKD.

  7. Organization and evolution of a gene-rich region of the mouse genome: a 12.7-Mb region deleted in the Del(13)Svea36H mouse.

    Science.gov (United States)

    Mallon, Ann-Marie; Wilming, Laurens; Weekes, Joseph; Gilbert, James G R; Ashurst, Jennifer; Peyrefitte, Sandrine; Matthews, Lucy; Cadman, Matthew; McKeone, Richard; Sellick, Chris A; Arkell, Ruth; Botcherby, Marc R M; Strivens, Mark A; Campbell, R Duncan; Gregory, Simon; Denny, Paul; Hancock, John M; Rogers, Jane; Brown, Steve D M

    2004-10-01

    Del(13)Svea36H (Del36H) is a deletion of approximately 20% of mouse chromosome 13 showing conserved synteny with human chromosome 6p22.1-6p22.3/6p25. The human region is lost in some deletion syndromes and is the site of several disease loci. Heterozygous Del36H mice show numerous phenotypes and may model aspects of human genetic disease. We describe 12.7 Mb of finished, annotated sequence from Del36H. Del36H has a higher gene density than the draft mouse genome, reflecting high local densities of three gene families (vomeronasal receptors, serpins, and prolactins) which are greatly expanded relative to human. Transposable elements are concentrated near these gene families. We therefore suggest that their neighborhoods are gene factories, regions of frequent recombination in which gene duplication is more frequent. The gene families show different proportions of pseudogenes, likely reflecting different strengths of purifying selection and/or gene conversion. They are also associated with relatively low simple sequence concentrations, which vary across the region with a periodicity of approximately 5 Mb. Del36H contains numerous evolutionarily conserved regions (ECRs). Many lie in noncoding regions, are detectable in species as distant as Ciona intestinalis, and therefore are candidate regulatory sequences. This analysis will facilitate functional genomic analysis of Del36H and provides insights into mouse genome evolution.

  8. Positive Youth Development, Life Satisfaction and Problem Behaviors of Adolescents in Intact and Non-Intact Families in Hong Kong

    Directory of Open Access Journals (Sweden)

    Daniel Tan Lei Shek

    2013-08-01

    Full Text Available This study investigated whether Chinese adolescents living in intact and non-intact families differed in their positive development, life satisfaction, and risk behavior. A total of 3,328 Secondary 1 students responded to measures of positive youth development (such as resilience and psychosocial competencies, life satisfaction, and risk behavior (substance abuse, delinquency, Internet addiction, consumption of pornographic materials, self-harm, and behavioral intention to engage in problem behavior. Findings revealed that adolescents growing up in intact families reported higher levels of positive developmental outcomes and life satisfaction as compared with adolescents from non-intact families. Adolescents in non-intact families also reported higher levels of risk behaviors than those growing up in intact families.

  9. 46 CFR 28.570 - Intact righting energy.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 1 2010-10-01 2010-10-01 false Intact righting energy. 28.570 Section 28.570 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY UNINSPECTED VESSELS REQUIREMENTS FOR COMMERCIAL FISHING INDUSTRY VESSELS Stability § 28.570 Intact righting energy. (a) Except as provided in paragraph (c) of...

  10. X-ray inspection for boreholes in intact trees

    NARCIS (Netherlands)

    Jansen, R.M.C.; Hemming, J.

    2010-01-01

    X-ray systems are commonly used for luggage inspection. The size of these systems is sufficient for inspection of intact trees. The first objective of this study is to determine whether such an X-ray system is able to visualise boreholes in intact trees. The present study is partly based on human in

  11. Development of spatial database on intact forest landscapes of India

    Science.gov (United States)

    Sudhakar Reddy, C.; Singh, Jyoti; Jha, C. S.; Diwakar, P. G.; Dadhwal, V. K.

    2017-01-01

    There is an increased interest in identifying the status of biodiversity in different spatial and temporal scales. The objective of the current research is to prepare a consistent spatial database of intact forest landscapes of India. The intact forest landscapes are located in the Himalayas, Andaman & Nicobar Islands, Western Ghats and Sunderbans. At national level 237 forest landscapes and 5.4% of the total natural forest remained as intact in India. Current intact forest landscapes of India consists of blocks larger than 10 km2 covering an area of 34,061 km2. Of the total area under intact forest landscapes, Eastern Himalayas represent 76.7% of the area, followed by Western Himalayas (8.8%), Andaman & Nicobar Islands (6.2%) and Western Ghats (5.7%). The largest intact forest landscape block occupies an area of 3342.9 km2 (9.8%) is found in western Arunachal Pradesh. Temperate forest zone represents the highest intactness (56.6%), followed by subtropical zone (19.2%), tropical zone (18.6%) and alpine zone (5.6%). Himalayan moist temperate forests represent the highest area (39.1%) of intact forest landscapes followed by subtropical broad-leaved hill forests, wet evergreen forests, and montane wet temperate forests. It is estimated that 4.4% of the area of intact forest landscapes fall inside the existing 47 protected areas. The results of the analysis best suited as input for the process of identification of new protected areas. The study recommends fine-scale mapping of biodiversity within the intact forest landscapes and to prepare efficient conservation plans.

  12. Vesta Is Not an Intact Protoplanet

    Science.gov (United States)

    Consolmagno, Guy; Turrini, Diego; Golabeck, Gregor; Jutzi, Martin; Sirono, Sin-iti; Svetsov, Vladimir; Tsiganis, Kleomenis

    2014-11-01

    Asteroid 4 Vesta has been identified as the likely source of howardite, eucrite, and diogenite (HED) basaltic achondrite meteorites, whose parent body differentiated and started solidifying within 3 Ma after the condensation of the Ca-Al-rich inclusions (CAIs). The formation of Jupiter and the disk-driven migration of the giant planets also occurred during this period; thus it was expected that Vesta could provide an intact record of large-scale early episodes of planetary migration and bombardment as in the proposed Jovian Early Bombardment and the “Grand Tack” scenarios. However, the results of the Dawn mission detailing Vesta’s mass, volume, density, and surface characteristics provide challenges for modeling the structure and evolution of this asteroid. All proposed models for the generation of the HEDs require the presence of a substantial olivine-rich mantle. But recent work on the depth of excavation of the large basins at the south pole of Vesta suggests that because there is not abundant mantle olivine visible on Vesta or in the Vestoid family asteroids, the crust of Vesta must be at least 80 km thick. Such a thick crust is radically at odds with previous models; should it exist, it ought to manifest itself in other ways such as Vesta’s density structure and bulk chemical composition. However, we find that no Vesta model of iron core, olivine-rich mantle, and HED crust can match the joint constraints of (a) Vesta’s density as derived from the gravity field observed by Dawn; (b) the observed depletion of sodium and potassium and trace element enrichments of the HED meteorites; and (c) the absence of exposed olivine on Vesta’s surface, among Vestoid asteroids, or in our collection of basaltic meteorites. Either Vesta was subjected to a radical change in composition, presumably due to the intense collisional environment where and when it formed, or the asteroid we see today is in fact a reaccretion of material formed elsewhere from now

  13. Mouse phenotyping.

    Science.gov (United States)

    Fuchs, Helmut; Gailus-Durner, Valérie; Adler, Thure; Aguilar-Pimentel, Juan Antonio; Becker, Lore; Calzada-Wack, Julia; Da Silva-Buttkus, Patricia; Neff, Frauke; Götz, Alexander; Hans, Wolfgang; Hölter, Sabine M; Horsch, Marion; Kastenmüller, Gabi; Kemter, Elisabeth; Lengger, Christoph; Maier, Holger; Matloka, Mikolaj; Möller, Gabriele; Naton, Beatrix; Prehn, Cornelia; Puk, Oliver; Rácz, Ildikó; Rathkolb, Birgit; Römisch-Margl, Werner; Rozman, Jan; Wang-Sattler, Rui; Schrewe, Anja; Stöger, Claudia; Tost, Monica; Adamski, Jerzy; Aigner, Bernhard; Beckers, Johannes; Behrendt, Heidrun; Busch, Dirk H; Esposito, Irene; Graw, Jochen; Illig, Thomas; Ivandic, Boris; Klingenspor, Martin; Klopstock, Thomas; Kremmer, Elisabeth; Mempel, Martin; Neschen, Susanne; Ollert, Markus; Schulz, Holger; Suhre, Karsten; Wolf, Eckhard; Wurst, Wolfgang; Zimmer, Andreas; Hrabě de Angelis, Martin

    2011-02-01

    Model organisms like the mouse are important tools to learn more about gene function in man. Within the last 20 years many mutant mouse lines have been generated by different methods such as ENU mutagenesis, constitutive and conditional knock-out approaches, knock-down, introduction of human genes, and knock-in techniques, thus creating models which mimic human conditions. Due to pleiotropic effects, one gene may have different functions in different organ systems or time points during development. Therefore mutant mouse lines have to be phenotyped comprehensively in a highly standardized manner to enable the detection of phenotypes which might otherwise remain hidden. The German Mouse Clinic (GMC) has been established at the Helmholtz Zentrum München as a phenotyping platform with open access to the scientific community (www.mousclinic.de; [1]). The GMC is a member of the EUMODIC consortium which created the European standard workflow EMPReSSslim for the systemic phenotyping of mouse models (http://www.eumodic.org/[2]). Copyright © 2010 Elsevier Inc. All rights reserved.

  14. Reflectance Spectra Comparison of Orbital Debris, Intact Spacecraft, and Intact Rocket Bodies in the GEO Regime

    Science.gov (United States)

    Albercromby, Kira J.; Abell, Paul; Barker, Ed

    2009-03-01

    A key objective of NASA's Orbital Debris program office at Johnson Space Center (JSC) is to characterize the debris environment by way of assessing the physical properties (type, mass, density, and size) of objects in orbit. Knowledge of the geosynchronous orbit (GEO) debris environment in particular can be used to determine the hazard probability at specific GEO altitudes and aid predictions of the future environment. To calculate an optical size from an intensity measurement of an object in the GEO regime, a 0.175 albedo is assumed currently. However, identification of specific material type or types could improve albedo accuracy and yield a more accurate size estimate for the debris piece. Using spectroscopy, it is possible to determine the surface materials of space objects. The study described herein used the NASA Infrared Telescope Facility (IRTF) to record spectral data in the ~ 0.65 to 2.5 micron regime on eight catalogued space objects. For comparison, all of the objects observed were in GEO or near-GEO. The eight objects consisted of two intact spacecraft, three rocket bodies, and three catalogued debris pieces. Two of the debris pieces stemmed from Titan 3C transtage breakup and the third is from COSMOS 2054. The reflectance spectra of the Titan 3C pieces share similar slopes (increasing with wavelength) and lack any strong absorption features. The COSMOS debris spectrum has a slight slope and has no absorption features. In contrast, the intact spacecraft show classic absorption features due to solar cells with a strong band gap feature near 1 micron. The two spacecraft were spin-stabilized objects and therefore have solar panels surrounding the outer surface. Two of the three rocket bodies are inertial upper stage (IUS) rocket bodies and have similar looking spectra. The slopes flatten out near 1.5 microns with absorption features in the near-infrared that are similar to that of white paint. The third rocket body has a similar flattening of slope but

  15. Treatment for burn blisters: debride or leave intact?

    Science.gov (United States)

    Murphy, Faye; Amblum, Jeshi

    2014-05-01

    This article presents findings from a systematic literature review of whether blisters arising from minor burns should be de-roofed or left intact. It discusses the risks of infection, healing outcomes, discomfort, choice of dressings and costs associated with each method, and reveals that debriding blisters larger than the patient's little fingernail while leaving smaller ones intact is generally agreed to be the best option. The article also explains external factors that influence the choice of whether to debride or leave blisters intact, reviews policy at the trust where one of the authors works in the context of the research and makes recommendations for practice.

  16. Vesta is not an intact protoplanet

    Science.gov (United States)

    Consolmagno, G.; Turrini, D.; Golabek, G.; Svetsov, V.; Sirono, S.; Tsiganis, K.

    2014-07-01

    The Dawn mission was designed to explore ''remnant intact protoplanets from the earliest epoch of solar system formation'' [1]. However, models of Vesta composed of an iron core, olivine mantle, and HED crust in chondritic proportions cannot match the joint constraints from Dawn [1] of Vesta's density, core size, and the extremely limited presence of exposed olivine on its surface. Vesta has a mean density of 3456 kg/m3 and its surface composition is well matched by howardites. The Dawn gravity data suggest a nickel-iron core of radius 110 km and density 7500--7800 kg/m3. The Rheasilvia impact basin, formed within a pre-existing large basin, Veneneia, should have excavated material from a depth of 50 km to 80 km or more below Vesta's surface [2]. If the howardite crust were thinner than 50--80 km, a significant amount of olivine-rich material, derived from depth, would have been exposed within this basin; models suggest that olivine would also be distributed both on Vesta's surface and in space as meteorite-source Vestoids. Such olivine is rare on Vesta, among the Vestoids, or in our meteorite collection. Vesta's density is similar to an L chondrite, but the Na and K abundances in Vesta are strongly depleted compared to chondrites and the average metal content of an L chondrite, 8.4% by mass, would give a core radius less than 90 km. A 110 km radius metallic core, via the Dawn data, represents 15% of Vesta's mass. The Mg/Al ratio in cosmic abundances is about 10:1, but roughly 1:1 within the eucrites; thus if Vesta started with cosmic abundances, the eucrites can only represent 10% of the parent body total mass. Likewise the 10 x chondritic rare earth trace elements (REE) abundance seen in most eucrites demands that, regardless of formation mechanism, these basalts were crystallized from a melt representing 10% of the mass of the source region [3]. Thus the howardite crust of a chondritic HED parent body, mixing all the available eucritic and diogenitic material

  17. Intact skin and not stripped skin is crucial for the safety and efficacy of peanut epicutaneous immunotherapy (EPIT in mice

    Directory of Open Access Journals (Sweden)

    Mondoulet Lucie

    2012-11-01

    Full Text Available Abstract Background Epicutaneous immunotherapy (EPIT on intact skin with an epicutaneous delivery system has already been used in preclinical and clinical studies. In epicutaneous vaccination and immunotherapy, the stripping of skin before application of the allergen is suggested to facilitate the passage of allergen through immune cells. Objectives The aim of this study was to compare the immunological response induced by EPIT performed on intact and stripped skin in a mouse model of peanut allergy. Methods After oral sensitization with peanut and cholera toxin, BALB/c mice were epicutaneously treated using an epicutaneous delivery system (Viaskin® (DBV Technologies, Paris applied either on intact skin or on stripped skin. Following EPIT, mice received an exclusive oral peanut regimen, aimed at triggering esophageal and jejunal lesions. We assessed eosinophil infiltration by histology, mRNA expression in the esophagus, antibody levels and peripheral T-cell response. Results EPIT on intact skin significantly reduced Th2 immunological response (IgE response and splenocyte secretion of Th2 cytokines as well as esophageal eosinophilia (2.7 ± 0.9, compared to Sham 19.9 ± 1.5, p  Conclusions Epicutaneous allergen-specific immunotherapy needs the integrity of superficial layers of the stratum corneum to warranty safety of treatment and to induce a tolerogenic profile of the immune response.

  18. NCBI nr-aa BLAST: CBRC-MMUS-06-0122 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0122 ref|NP_444458.1| vomeronasal 1 receptor, B7 [Mus musculus] sp|Q9EQ46|VN1B7_MOUSE Vomeronas...al type-1 receptor B7 (Vomeronasal type-1 receptor A11) gb|AAG42087.1|AF291493_1 vomeronasa...l receptor V1RB7 [Mus musculus] dbj|BAB79217.1| vomeronasal receptor 1 A11 [Mus musculus] gb|AAI41627.1| Vomeronasa...l 1 receptor, B7 [synthetic construct] gb|AAI40251.1| Vomeronasal

  19. NCBI nr-aa BLAST: CBRC-MMUS-06-0135 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0135 ref|NP_444458.1| vomeronasal 1 receptor, B7 [Mus musculus] sp|Q9EQ46|VN1B7_MOUSE Vomeronas...al type-1 receptor B7 (Vomeronasal type-1 receptor A11) gb|AAG42087.1|AF291493_1 vomeronasa...l receptor V1RB7 [Mus musculus] dbj|BAB79217.1| vomeronasal receptor 1 A11 [Mus musculus] gb|AAI41627.1| Vomeronasa...l 1 receptor, B7 [synthetic construct] gb|AAI40251.1| Vomeronasal

  20. NCBI nr-aa BLAST: CBRC-MMUS-06-0123 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0123 ref|NP_444458.1| vomeronasal 1 receptor, B7 [Mus musculus] sp|Q9EQ46|VN1B7_MOUSE Vomeronas...al type-1 receptor B7 (Vomeronasal type-1 receptor A11) gb|AAG42087.1|AF291493_1 vomeronasa...l receptor V1RB7 [Mus musculus] dbj|BAB79217.1| vomeronasal receptor 1 A11 [Mus musculus] gb|AAI41627.1| Vomeronasa...l 1 receptor, B7 [synthetic construct] gb|AAI40251.1| Vomeronasal

  1. NCBI nr-aa BLAST: CBRC-EEUR-01-0634 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-EEUR-01-0634 ref|NP_444458.1| vomeronasal 1 receptor, B7 [Mus musculus] sp|Q9EQ46|VN1B7_MOUSE Vomeronas...al type-1 receptor B7 (Vomeronasal type-1 receptor A11) gb|AAG42087.1|AF291493_1 vomeronasa...l receptor V1RB7 [Mus musculus] dbj|BAB79217.1| vomeronasal receptor 1 A11 [Mus musculus] gb|AAI41627.1| Vomeronasa...l 1 receptor, B7 [synthetic construct] gb|AAI40251.1| Vomeronasal

  2. NCBI nr-aa BLAST: CBRC-MMUS-06-0130 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0130 ref|NP_444448.1| vomeronasal 1 receptor, A3 [Mus musculus] sp|Q9EQ52|VN1A3_MOUSE Vomeronas...al type-1 receptor A3 (Vomeronasal type-1 receptor A6) gb|AAG42076.1|AF291482_1 vomeronasal... receptor V1RA3 [Mus musculus] dbj|BAB79212.1| vomeronasal receptor 1 A6 [Mus musculus] gb|AAI41612.1| Vomeronasal... 1 receptor, A3 [synthetic construct] gb|AAI40226.1| Vomeronasal 1 receptor, A3 [synthetic construct] NP_444448.1 1e-144 88% ...

  3. NCBI nr-aa BLAST: CBRC-OCUN-01-1320 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OCUN-01-1320 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal

  4. NCBI nr-aa BLAST: CBRC-MMUS-06-0139 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0139 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal

  5. NCBI nr-aa BLAST: CBRC-OCUN-01-0928 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OCUN-01-0928 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal

  6. NCBI nr-aa BLAST: CBRC-RNOR-04-0244 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-04-0244 ref|NP_444458.1| vomeronasal 1 receptor, B7 [Mus musculus] sp|Q9EQ46|VN1B7_MOUSE Vomeronas...al type-1 receptor B7 (Vomeronasal type-1 receptor A11) gb|AAG42087.1|AF291493_1 vomeronasal... receptor V1RB7 [Mus musculus] dbj|BAB79217.1| vomeronasal receptor 1 A11 [Mus musculus] gb|AAI41627.1| Vomeronasal... 1 receptor, B7 [synthetic construct] gb|AAI40251.1| Vomeronasal

  7. NCBI nr-aa BLAST: CBRC-MMUS-06-0133 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0133 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal

  8. NCBI nr-aa BLAST: CBRC-RNOR-04-0249 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-04-0249 ref|NP_444458.1| vomeronasal 1 receptor, B7 [Mus musculus] sp|Q9EQ46|VN1B7_MOUSE Vomeronas...al type-1 receptor B7 (Vomeronasal type-1 receptor A11) gb|AAG42087.1|AF291493_1 vomeronasal... receptor V1RB7 [Mus musculus] dbj|BAB79217.1| vomeronasal receptor 1 A11 [Mus musculus] gb|AAI41627.1| Vomeronasal... 1 receptor, B7 [synthetic construct] gb|AAI40251.1| Vomeronasal

  9. NCBI nr-aa BLAST: CBRC-OCUN-01-0037 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OCUN-01-0037 ref|NP_444448.1| vomeronasal 1 receptor, A3 [Mus musculus] sp|Q9EQ52|VN1A3_MOUSE Vomeronas...al type-1 receptor A3 (Vomeronasal type-1 receptor A6) gb|AAG42076.1|AF291482_1 vomeronasal... receptor V1RA3 [Mus musculus] dbj|BAB79212.1| vomeronasal receptor 1 A6 [Mus musculus] gb|AAI41612.1| Vomeronasal... 1 receptor, A3 [synthetic construct] gb|AAI40226.1| Vomeronasal 1 receptor, A3 [synthetic construct] NP_444448.1 5e-76 48% ...

  10. NCBI nr-aa BLAST: CBRC-MMUS-06-0136 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0136 ref|NP_444458.1| vomeronasal 1 receptor, B7 [Mus musculus] sp|Q9EQ46|VN1B7_MOUSE Vomeronas...al type-1 receptor B7 (Vomeronasal type-1 receptor A11) gb|AAG42087.1|AF291493_1 vomeronasal... receptor V1RB7 [Mus musculus] dbj|BAB79217.1| vomeronasal receptor 1 A11 [Mus musculus] gb|AAI41627.1| Vomeronasal... 1 receptor, B7 [synthetic construct] gb|AAI40251.1| Vomeronasal

  11. NCBI nr-aa BLAST: CBRC-RNOR-04-0240 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-04-0240 ref|NP_444458.1| vomeronasal 1 receptor, B7 [Mus musculus] sp|Q9EQ46|VN1B7_MOUSE Vomeronas...al type-1 receptor B7 (Vomeronasal type-1 receptor A11) gb|AAG42087.1|AF291493_1 vomeronasal... receptor V1RB7 [Mus musculus] dbj|BAB79217.1| vomeronasal receptor 1 A11 [Mus musculus] gb|AAI41627.1| Vomeronasal... 1 receptor, B7 [synthetic construct] gb|AAI40251.1| Vomeronasal

  12. NCBI nr-aa BLAST: CBRC-MMUS-06-0134 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0134 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal

  13. NCBI nr-aa BLAST: CBRC-OCUN-01-0037 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OCUN-01-0037 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal

  14. Analysis of 16S libraries of mouse gastrointestinal microflora reveals a large new group of mouse intestinal bacteria

    NARCIS (Netherlands)

    Salzman, NH; de Jong, H; Paterson, Y; Harmsen, HJM; Welling, GW; Bos, NA

    2002-01-01

    Total genomic DNA from samples of intact mouse small intestine, large intestine, caecum and faeces was used as template for PCR amplification of 16S rRNA gene sequences with conserved bacterial primers. Phylogenetic analysis of the amplification products revealed 40 unique 16S rDNA sequences. Of

  15. Subcellular distribution of glycogen and decreased tetanic Ca2+ in fatigued single intact mouse muscle fibres

    DEFF Research Database (Denmark)

    Nielsen, Joachim; Cheng, Arthur J; Ørtenblad, Niels

    2014-01-01

    , 350 ms tetani given at 2 s (high-intensity fatigue, HIF) or 10 s (low-intensity fatigue, LIF) intervals, while force and [Ca(2+)]i were measured. Stimulation continued until force decreased to 30% of its initial value. Fibres were then prepared for analyses of subcellular glycogen distribution...

  16. Blood flow dependence of the intratumoral distribution of peripheral benzodiazepine receptor binding in intact mouse fibrosarcoma

    Energy Technology Data Exchange (ETDEWEB)

    Amitani, Misato [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan) and Course of Allied Health Sciences, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871 (Japan)]. E-mail: amitani@sahs.med.osaka-u.ac.jp; Zhang, Ming-Rong [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); Noguchi, Junko [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); SHI Accelerator Service, Shinagawa-ku, Tokyo 141-8686 (Japan); Kumata, Katsushi [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); Ito, Takehito [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); SHI Accelerator Service, Shinagawa-ku, Tokyo 141-8686 (Japan); Takai, Nobuhiko [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); Suzuki, Kazutoshi [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); Hosoi, Rie [Course of Allied Health Sciences, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871 (Japan); Inoue, Osamu [Course of Allied Health Sciences, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871 (Japan)

    2006-11-15

    The intratumoral distribution of [{sup 11}C]AC-5216 binding, a novel peripheral benzodiazepine receptor (PBR) ligand, was examined by autoradiography both in vitro and in vivo using a murine fibrosarcoma model. The regional distribution of [{sup 11}C]AC-5216 in a tumor in vivo was significantly heterogeneous; the uptake of [{sup 11}C]AC-5216 was comparatively higher in the outer rim of the tumor and was lower in the central area. In contrast, the images obtained following the injection of [{sup 11}C]AC-5216 with a large amount of nonlabeled PK11195 showed a relatively homogeneous distribution, suggesting that [{sup 11}C]AC-5216 uptake represented specific binding to PBRs. In vitro autoradiograms of [{sup 11}C]AC-5216 binding were also obtained using the section of the fibrosarcoma that was the same as that used to examine in vivo binding. In vitro autoradiographic binding images showed homogeneous distribution, and significant discrepancies of the intratumoral distribution of [{sup 11}C]AC-5216 were observed between in vivo and in vitro images. The in vivo images of [{sup 11}C]AC-5216 uptake, compared with those of [{sup 14}C]iodoantipyrine uptake, obtained by dual autoradiography to evaluate the influence of blood flow revealed the similar intratumoral distributions of both tracers. These results indicate that the delivery process from the plasma to the tumor might be the rate-limiting step for the intratumoral distribution of PBR binding in vivo in a fibrosarcoma model.

  17. In vivo robotics: the automation of neuroscience and other intact-system biological fields.

    Science.gov (United States)

    Kodandaramaiah, Suhasa B; Boyden, Edward S; Forest, Craig R

    2013-12-01

    Robotic and automation technologies have played a huge role in in vitro biological science, having proved critical for scientific endeavors such as genome sequencing and high-throughput screening. Robotic and automation strategies are beginning to play a greater role in in vivo and in situ sciences, especially when it comes to the difficult in vivo experiments required for understanding the neural mechanisms of behavior and disease. In this perspective, we discuss the prospects for robotics and automation to influence neuroscientific and intact-system biology fields. We discuss how robotic innovations might be created to open up new frontiers in basic and applied neuroscience and present a concrete example with our recent automation of in vivo whole-cell patch clamp electrophysiology of neurons in the living mouse brain.

  18. Mouse models for methylmalonic aciduria.

    Directory of Open Access Journals (Sweden)

    Heidi L Peters

    Full Text Available Methylmalonic aciduria (MMA is a disorder of organic acid metabolism resulting from a functional defect of methylmalonyl-CoA mutase (MCM. MMA is associated with significant morbidity and mortality, thus therapies are necessary to help improve quality of life and prevent renal and neurological complications. Transgenic mice carrying an intact human MCM locus have been produced. Four separate transgenic lines were established and characterised as carrying two, four, five or six copies of the transgene in a single integration site. Transgenic mice from the 2-copy line were crossed with heterozygous knockout MCM mice to generate mice hemizygous for the human transgene on a homozygous knockout background. Partial rescue of the uniform neonatal lethality seen in homozygous knockout mice was observed. These rescued mice were significantly smaller than control littermates (mice with mouse MCM gene. Biochemically, these partial rescue mice exhibited elevated methylmalonic acid levels in urine, plasma, kidney, liver and brain tissue. Acylcarnitine analysis of blood spots revealed elevated propionylcarnitine levels. Analysis of mRNA expression confirms the human transgene is expressed at higher levels than observed for the wild type, with highest expression in the kidney followed closely by brain and liver. Partial rescue mouse fibroblast cultures had only 20% of the wild type MCM enzyme activity. It is anticipated that this humanised partial rescue mouse model of MMA will enable evaluation of long-term pathophysiological effects of elevated methylmalonic acid levels and be a valuable model for the investigation of therapeutic strategies, such as cell transplantation.

  19. Mouse Models for Methylmalonic Aciduria

    Science.gov (United States)

    Peters, Heidi L.; Pitt, James J.; Wood, Leonie R.; Hamilton, Natasha J.; Sarsero, Joseph P.; Buck, Nicole E.

    2012-01-01

    Methylmalonic aciduria (MMA) is a disorder of organic acid metabolism resulting from a functional defect of methylmalonyl-CoA mutase (MCM). MMA is associated with significant morbidity and mortality, thus therapies are necessary to help improve quality of life and prevent renal and neurological complications. Transgenic mice carrying an intact human MCM locus have been produced. Four separate transgenic lines were established and characterised as carrying two, four, five or six copies of the transgene in a single integration site. Transgenic mice from the 2-copy line were crossed with heterozygous knockout MCM mice to generate mice hemizygous for the human transgene on a homozygous knockout background. Partial rescue of the uniform neonatal lethality seen in homozygous knockout mice was observed. These rescued mice were significantly smaller than control littermates (mice with mouse MCM gene). Biochemically, these partial rescue mice exhibited elevated methylmalonic acid levels in urine, plasma, kidney, liver and brain tissue. Acylcarnitine analysis of blood spots revealed elevated propionylcarnitine levels. Analysis of mRNA expression confirms the human transgene is expressed at higher levels than observed for the wild type, with highest expression in the kidney followed closely by brain and liver. Partial rescue mouse fibroblast cultures had only 20% of the wild type MCM enzyme activity. It is anticipated that this humanised partial rescue mouse model of MMA will enable evaluation of long-term pathophysiological effects of elevated methylmalonic acid levels and be a valuable model for the investigation of therapeutic strategies, such as cell transplantation. PMID:22792386

  20. Intact skull chronic windows for mesoscopic wide-field imaging in awake mice

    Science.gov (United States)

    Silasi, Gergely; Xiao, Dongsheng; Vanni, Matthieu P.; Chen, Andrew C. N.; Murphy, Timothy H.

    2016-01-01

    Background Craniotomy-based window implants are commonly used for microscopic imaging, in head-fixed rodents, however their field of view is typically small and incompatible with mesoscopic functional mapping of cortex. New Method We describe a reproducible and simple procedure for chronic through-bone wide-field imaging in awake head-fixed mice providing stable optical access for chronic imaging over large areas of the cortex for months. Results The preparation is produced by applying clear-drying dental cement to the intact mouse skull, followed by a glass coverslip to create a partially transparent imaging surface. Surgery time takes about 30 minutes. A single set-screw provides a stable means of attachment for mesoscale assessment without obscuring the cortical field of view. Comparison with Existing Methods We demonstrate the utility of this method by showing seed-pixel functional connectivity maps generated from spontaneous cortical activity of GCAMP6 signals in both awake and anesthetized mice. Conclusions We propose that the intact skull preparation described here may be used for most longitudinal studies that do not require micron scale resolution and where cortical neural or vascular signals are recorded with intrinsic sensors. PMID:27102043

  1. In Vivo Optical Imaging of Neurogenesis: Watching New Neurons in the Intact Brain

    Directory of Open Access Journals (Sweden)

    Sebastien Couillard-Despres

    2008-01-01

    Full Text Available Adult neurogenesis is a highly dynamic process modulated by several pathologic and environmental factors, as well as by various compounds. So far, available techniques to study neurogenesis are lengthy and personnel and cost intensive. We developed a new tool based on the doublecortin promoter driving the expression of the luciferase reporter gene (DCX-promo-luciferase in transgenic mice to perform in vivo imaging of neurogenesis. Indeed, the DCX-promo-luciferase mice allowed optical in vivo imaging of the onset of and increase in neurogenesis in developing fetal brains, as well as imaging of neurogenesis in the intact adult mouse central nervous system. Moreover, the capacity to specifically detect a small number of migrating neuronal precursors in vivo after transplantation is for the first time feasible using this DCX-promo-luciferase transgenic tool. The present imaging approach offers several crucial advantages over methods currently available, such as bromodeoxyuridine incorporation or labeling using iron oxide nanoparticles. Hence, it allows longitudinal study of neurogenesis in intact animals without the requirement of cellular prelabeling. Moreover, it guarantees that detection is specific for neuronal precursors and restricted to viable cells. Hence, our DCX-promo-luciferase transgenic model constitutes an effective tool that answers the pressing need for rapid investigation of the impact on neurogenesis of a large number of candidate compounds waiting to be tested.

  2. Sexual incentive motivation, olfactory preference, and activation of the vomeronasal projection pathway by sexually relevant cues in non-copulating and naive male rats.

    Science.gov (United States)

    Portillo, Wendy; Paredes, Raúl G

    2004-09-01

    There are some apparently healthy male rats that fail to mate after repeated testing with receptive females. We have previously shown that these "non-copulator (NC)" males show no partner preference for a receptive female when given the opportunity to physically interact with a sexually receptive female or a sexually active male. We also demonstrated that although NC males prefer odors from estrous females to odors from anestrous females, this preference is significantly reduced in comparison to the preference displayed by copulating (C) males. The aim of the present study was to evaluate in NC males sexual incentive motivation, that is, the approach behavior of male rats to either a sexually receptive female or a sexually active male in a test where the subjects can smell, hear, and see the stimulus animal but prevents their physical interaction. In addition, we determined whether NC rats have alterations in their ability to detect odors from conspecifics or odors related to food. In the detection of odors from conspecifics, we determined if these NC males are sexually attracted toward odors from receptive females or sexually active males. For food-related odors, we quantified the time it took the subjects to locate a hidden a piece of apple. Finally, using the induction of Fos-immunoreactivity (Fos-IR) as an index of neuronal activation, we compared the response of the vomeronasal projection pathway (VN pathway) of C and NC male rats exposed to estrous bedding. Males without sexual experience (WSE) were included in all experiments to determine the importance of previous heterosexual experience in the different behavioral tests and in the activity of the VN pathway. In the sexual incentive motivation test, we found that C and WSE male rats have a clear preference for estrous females over sexually active males, whereas NC male rats showed no preference. In odor tests, our results showed that C males had a clear preference for odors from estrous females as opposed

  3. Vulnerability of ecosystems to climate change moderated by habitat intactness.

    Science.gov (United States)

    Eigenbrod, Felix; Gonzalez, Patrick; Dash, Jadunandan; Steyl, Ilse

    2015-01-01

    The combined effects of climate change and habitat loss represent a major threat to species and ecosystems around the world. Here, we analyse the vulnerability of ecosystems to climate change based on current levels of habitat intactness and vulnerability to biome shifts, using multiple measures of habitat intactness at two spatial scales. We show that the global extent of refugia depends highly on the definition of habitat intactness and spatial scale of the analysis of intactness. Globally, 28% of terrestrial vegetated area can be considered refugia if all natural vegetated land cover is considered. This, however, drops to 17% if only areas that are at least 50% wilderness at a scale of 48×48 km are considered and to 10% if only areas that are at least 50% wilderness at a scale of 4.8×4.8 km are considered. Our results suggest that, in regions where relatively large, intact wilderness areas remain (e.g. Africa, Australia, boreal regions, South America), conservation of the remaining large-scale refugia is the priority. In human-dominated landscapes, (e.g. most of Europe, much of North America and Southeast Asia), focusing on finer scale refugia is a priority because large-scale wilderness refugia simply no longer exist. Action to conserve such refugia is particularly urgent since only 1 to 2% of global terrestrial vegetated area is classified as refugia and at least 50% covered by the global protected area network. © 2014 John Wiley & Sons Ltd.

  4. Persistent truncus arteriosus with intact ventricular septum diagnosed by echocardiography

    Institute of Scientific and Technical Information of China (English)

    ZHANG Yu-qi; SHEN Rong; SUN Kun; ZHONG Shu-wen; WU Yu-rong

    2009-01-01

    Persistent truncus arteriosus (PTA) is a rare congenital cardiac anomaly, and has an incidence of about 0.5 to 0.9 per 10 000 live births. Almost all cases described in the literatures had a large ventricular septal defect, only few rare cases were reported with intact ventricular septum. From June 1998 to December 2008, cardiac angiography were performed in 10 880 patients with congenital heart disease in our hospital, 47 patients with PTA were diagnosed, one case with tricuspid atresia,hypoplastic right ventricle, and intact ventricular septum was encountered.

  5. Perceived parental control processes, parent-child relational qualities and psychological well-being of Chinese adolescents in intact and non-intact families in Hong Kong.

    Science.gov (United States)

    Shek, Daniel T L; Lee, Tak Yan

    2007-01-01

    This paper examines whether Chinese adolescents' perceptions (N = 3,017) of parental behavioral control (parental knowledge, expectation, monitoring, discipline, and demandingness as well as parental control based on indigenous Chinese concepts), parental psychological control, parent-child relational qualities (perceived parental trust, child's trust of the parents, child's readiness to communicate with the parents, and child's satisfaction with parental control), and adolescent psychological well-being (hopelessness, mastery, life satisfaction and self-esteem) differed in intact and non-intact families. Results showed that relative to non-intact families, parental behavioral control processes were higher and parent-child relational qualities were better in intact families. In contrast, parental psychological control was higher in non-intact families than in intact families. Finally, the psychological well-being of adolescents in non-intact families was poorer than that of adolescents in intact families.

  6. Bioavailability and in vivo metabolism of intact glucosinolates

    DEFF Research Database (Denmark)

    Sørensen, Jens Christian; Frandsen, Heidi Blok; Jensen, Søren Krogh

    2016-01-01

    Health benefits associated with consumption of cruciferous vegetables have received considerable attention with a hitherto focus on the role and bioactivity of glucosinolate degradation products. We investigated the in vivo metabolism of intact glucosinolates by following their fate in digesta an...

  7. Fostering Activities of Daily Living by Intact Nursing Home Residents

    Science.gov (United States)

    Blair, Charles E.; Glaister, Judy; Brown, Alston; Phillips, Carolyn

    2007-01-01

    We assessed effectiveness of four education programs in providing nursing assistants with ability to produce a therapeutic milieu supportive of intact residents' activities of daily living, positive self-esteem and mood: (1) a combination of Orem's Systems of Nursing Care and Skinner's Applied Behavioral Analysis, (2) Applied Behavioral Analysis,…

  8. Cholesteric carbohydrate liquid crystals incorporating an intact glucopyranose moiety

    NARCIS (Netherlands)

    Smits, E; Engberts, J.B.F.N.; Kellogg, R.M; van Doren, H.A.

    1997-01-01

    Recently, the first monosaccharide derivatives containing a fully intact monosaccharide and two vicinal OH-groups which display thermotropic chiral mesophases were synthesized. These liquid crystals have a rigid core, with a trans-decalin-like skeleton incorporating the D-glucopyranose ring, substit

  9. In amnio MRI of mouse embryos.

    Directory of Open Access Journals (Sweden)

    Thomas A Roberts

    Full Text Available Mouse embryo imaging is conventionally carried out on ex vivo embryos excised from the amniotic sac, omitting vital structures and abnormalities external to the body. Here, we present an in amnio MR imaging methodology in which the mouse embryo is retained in the amniotic sac and demonstrate how important embryonic structures can be visualised in 3D with high spatial resolution (100 µm/px. To illustrate the utility of in amnio imaging, we subsequently apply the technique to examine abnormal mouse embryos with abdominal wall defects. Mouse embryos at E17.5 were imaged and compared, including three normal phenotype embryos, an abnormal embryo with a clear exomphalos defect, and one with a suspected gastroschisis phenotype. Embryos were excised from the mother ensuring the amnion remained intact and stereo microscopy was performed. Embryos were next embedded in agarose for 3D, high resolution MRI on a 9.4T scanner. Identification of the abnormal embryo phenotypes was not possible using stereo microscopy or conventional ex vivo MRI. Using in amnio MRI, we determined that the abnormal embryos had an exomphalos phenotype with varying severities. In amnio MRI is ideally suited to investigate the complex relationship between embryo and amnion, together with screening for other abnormalities located outside of the mouse embryo, providing a valuable complement to histology and existing imaging methods available to the phenotyping community.

  10. Pharmacokinetics of erythropoietin in intact and anephric dogs

    Energy Technology Data Exchange (ETDEWEB)

    Fu, J.S.; Lertora, J.J.; Brookins, J.; Rice, J.C.; Fisher, J.W.

    1988-06-01

    The present studies were performed to determine the pharmacokinetic parameters of erythropoietin in intact and anephric dogs by use of unlabeled crude native erythropoietin (nEp) and iodine 125-labeled purified recombinant erythropoietin (rEp) given by intravenous infusion for 15 minutes. Sephadex G-75 gel filtration was used to confirm that the 125I-rEp molecule remained iodinated in dog plasma during the 24-hour period of these studies. The plasma disappearance of erythropoietin conformed to a biexponential equation for both nEp and 125I-rEp, with the central compartment being larger than the peripheral compartment. The mean distribution half-life of 75.3 +/- 21.2 minutes for nEp was significantly (p less than 0.05) longer than that of 125I-rEp (23.7 +/- 5.0 minutes) in intact dogs. The intercompartmental clearance (CIic) for nEp (0.018 +/- 0.006 L/kg/hr) was significantly smaller than that of 125I-rEp (0.068 +/- 0.018 L/kg/hr) in intact dogs (p less than 0.05). There were no significant differences in apparent volume of distribution, elimination half-life, and elimination clearance (CIe) for nEp and rEp in intact dogs. The mean elimination half-life for 125I-rEp in intact dogs (9.0 +/- 0.6 hours) and anephric dogs (13.8 +/- 1.4 hours) was significantly different (p less than 0.05). The CIe for 125I-rEp in anephric dogs (0.008 +/- 0.001 L/kg/hr) was significantly (p less than 0.05) smaller than that of 125I-rEp in intact dogs (0.011 +/- 0.001 L/kg/hr). There were no significant differences in apparent volume of distribution, distribution half-life, and CIic for 125I-rEp in intact and anephric dogs.

  11. Threats to intact tropical peatlands and opportunities for their conservation.

    Science.gov (United States)

    Roucoux, K H; Lawson, I T; Baker, T R; Del Castillo Torres, D; Draper, F C; Lähteenoja, O; Gilmore, M P; Honorio Coronado, E N; Kelly, T J; Mitchard, E T A; Vriesendorp, C F

    2017-03-08

    Large, intact areas of tropical peatland are highly threatened at a global scale by the expansion of commercial agriculture and other forms of economic development. Conserving peatlands on a landscape scale, with their hydrology intact, is of international conservation importance to preserve their distinctive biodiversity and ecosystem services and maintain their resilience to future environmental change. We explored threats to and opportunities for conserving remaining intact tropical peatlands; thus, we excluded peatlands of Indonesia and Malaysia, where extensive deforestation, drainage, and conversion to plantations means conservation in this region can protect only small fragments of the original ecosystem. We focused on a case study, the Pastaza-Marañón Foreland Basin (PMFB) in Peru, which is among the largest known intact tropical peatland landscapes in the world and is representative of peatland vulnerability. Maintenance of the hydrological conditions critical for carbon storage and ecosystem function of peatlands is, in the PMFB, primarily threatened by expansion of commercial agriculture linked to new transport infrastructure that is facilitating access to remote areas. There remain opportunities in the PMFB and elsewhere to develop alternative, more sustainable land-use practices. Although some of the peatlands in the PMFB fall within existing legally protected areas, this protection does not include the most carbon-dense (domed pole forest) areas. New carbon-based conservation instruments (e.g., REDD+, Green Climate Fund), developing markets for sustainable peatland products, transferring land title to local communities, and expanding protected areas offer pathways to increased protection for intact tropical peatlands in Amazonia and elsewhere, such as those in New Guinea and Central Africa which remain, for the moment, broadly beyond the frontier of commercial development. © 2017 The Authors. Conservation Biology published by Wiley Periodicals, Inc

  12. The Multilevel Mixed Intact Group Analysis: A Mixed Method to Seek, Detect, Describe, and Explain Differences Among Intact Groups

    Science.gov (United States)

    Schoonenboom, Judith

    2016-01-01

    Educational innovations often involve intact subgroups, such as school classes or university departments. In small-scale educational evaluation research, typically involving 1 to 20 subgroups, differences among these subgroups are often neglected. This article presents a mixed method from a qualitative perspective, in which differences among…

  13. Spelling-stress regularity effects are intact in developmental dyslexia.

    Science.gov (United States)

    Mundy, Ian R; Carroll, Julia M

    2013-01-01

    The current experiment investigated conflicting predictions regarding the effects of spelling-stress regularity on the lexical decision performance of skilled adult readers and adults with developmental dyslexia. In both reading groups, lexical decision responses were significantly faster and significantly more accurate when the orthographic structure of a word ending was a reliable as opposed to an unreliable predictor of lexical stress assignment. Furthermore, the magnitude of this spelling-stress regularity effect was found to be equivalent across reading groups. These findings are consistent with intact phoneme-level regularity effects also observed in dyslexia. The paper discusses how findings of intact spelling-sound regularity effects at both prosodic and phonemic levels, as well as other similar results, can be reconciled with the obvious difficulties that people with dyslexia experience in other domains of phonological processing.

  14. Structural determination of intact proteins using mass spectrometry

    Science.gov (United States)

    Kruppa, Gary; Schoeniger, Joseph S.; Young, Malin M.

    2008-05-06

    The present invention relates to novel methods of determining the sequence and structure of proteins. Specifically, the present invention allows for the analysis of intact proteins within a mass spectrometer. Therefore, preparatory separations need not be performed prior to introducing a protein sample into the mass spectrometer. Also disclosed herein are new instrumental developments for enhancing the signal from the desired modified proteins, methods for producing controlled protein fragments in the mass spectrometer, eliminating complex microseparations, and protein preparatory chemical steps necessary for cross-linking based protein structure determination.Additionally, the preferred method of the present invention involves the determination of protein structures utilizing a top-down analysis of protein structures to search for covalent modifications. In the preferred method, intact proteins are ionized and fragmented within the mass spectrometer.

  15. Binding of Clostridium botulinum C3 exoenzyme to intact cells.

    Science.gov (United States)

    Rohrbeck, Astrid; von Elsner, Leonie; Hagemann, Sandra; Just, Ingo

    2014-06-01

    C3 from Clostridium botulinum (C3) specifically modifies Rho GTPases RhoA, RhoB, and RhoC by mono-ADP-ribosylation. The confined substrate profile of C3 is the basis for its use as pharmacological tool in cell biology to study cellular functions of Rho GTPases. Although C3 exoenzyme does not possess a cell-binding/-translocation domain, C3 is taken up by intact cells via an unknown mechanism. In the present work, binding of C3 to the hippocampus-derived HT22 cells and J774A.1 macrophages was characterized. C3 bound concentration-dependent to HT22 and J774A.1 cells. Pronase treatment of intact cells significantly reduced both C3 binding and C3 cell entry. Removal of sugar residues by glycosidase F treatment resulted in an increased binding of C3, but a reduced cell entry. To explore the involvement of phosphorylation in the binding process of C3, intact HT22 and J774A.1 cells were pre-treated with vanadate prior to incubation with C3. Inhibition of de-phosphorylation by vanadate resulted in an increased binding of C3. To differentiate between intracellular and extracellular phosphorylation, intact cells were treated with CIP (calf intestine phosphatase) to remove extracellular phosphate residues. The removal of phosphate residues resulted in a strong reduction in binding of C3 to cells. In sum, the C3 membranous binding partner is proteinaceous, and the glycosylation as well as the phosphorylation state is critical for efficient binding of C3.

  16. Crisscross heart with dextrocardia and intact interventricular septum

    Directory of Open Access Journals (Sweden)

    P Kader Muneer

    2014-01-01

    Full Text Available Crisscross heart is a rare congenital heart disease characterized by a twisted atrioventricular connection, as a result of rotation of the ventricular mass along its long axis. We report an asymptomatic 48-year-old woman referred to us for evaluation of a cardiac murmur. Further evaluation showed situs solitus, dextrocardia with normal atrioventricular and ventriculoarterial connection, and a crisscross relation of the atrioventricular valves. Unlike the usual case of crisscross heart, our patient had an intact ventricular septum.

  17. The intact urokinase receptor is required for efficient vitronectin binding

    DEFF Research Database (Denmark)

    Høyer-Hansen, G; Behrendt, N; Ploug, M;

    1997-01-01

    The urokinase receptor (uPAR) is a receptor for both urokinase plasminogen activator (uPA) and the adhesion protein vitronectin. There are two forms of cell surface-bound uPAR; intact uPAR and a cleaved form, uPAR(2+3), which is formed by uPA-catalyzed cleavage of uPAR. In ligand-blotting experim...

  18. Pancreatic exocrine studies in intact animals: historic and current methods.

    Science.gov (United States)

    Niebergall-Roth, E; Teyssen, S; Singer, M V

    1997-12-01

    This report presents a review of the historic and current methods for performing pancreatic exocrine studies in intact animals. Special emphasis is given to the various surgical procedures--pancreatic fistulas, duodenal pouches, and duodenal fistulas--and practice of collecting pancreatic secretion in dogs. Procedures in other animal species--rat, cat, pig, rabbit, cattle, sheep, and horse--also are specified. The advantages and disadvantages, as well as the indications and limitations of the distinct methods, are discussed.

  19. Comparison of the stability of split and intact gabapentin tablets.

    Science.gov (United States)

    Volpe, Donna A; Gupta, Abhay; Ciavarella, Anthony B; Faustino, Patrick J; Sayeed, Vilayat A; Khan, Mansoor A

    2008-02-28

    The purpose of this study was to determine the stability differences between split and intact gabapentin tablets. Gabapentin tablets from three different manufacturers (G1, G2 and G3) were tested for a period of 9 weeks under long-term (25 degrees C/60% RH) and intermediate stability (30 degrees C/60% RH) storage conditions after storage in closed amber pharmacy dispensing containers. Samples were analyzed for dissolution and potency using validated HPLC methods. Potency test also included the quantitation of gabapentin's main degradation product. Tablets from all manufacturers and at all time points had potency >90%. At 9 weeks, a statistically significant decrease (psplit G2 and G3 tablets under the intermediate storage conditions. At the end of 9 weeks, all samples also showed slightly higher levels of degradation product which was statistically significant (psplit and intact tablets. No difference was observed between the potency and dissolution of the intact and the split tablets from the same manufacturer and the three products tested remained stable throughout the study period. The results suggest that splitting of gabapentin tablets did not affect the stability of these particular drug products tested as part of this study when stored under normal storage conditions for a period of up to 9 weeks. However, the results should not be extrapolated to other gabapentin drug products and to other tablet dosage forms.

  20. Does human leukocyte elastase degrade intact skin elastin?

    Science.gov (United States)

    Schmelzer, Christian E H; Jung, Michael C; Wohlrab, Johannes; Neubert, Reinhard H H; Heinz, Andrea

    2012-11-01

    This study aimed to investigate the susceptibility of intact fibrillar human elastin to human leukocyte elastase and cathepsin G. Elastin is a vital protein of the extracellular matrix of vertebrates, and provides exceptional properties including elasticity and tensile strength to many tissues and organs, including the aorta, lung, cartilage, elastic ligaments and skin, and is thus critical for their long-term function. Mature elastin is an insoluble and extremely durable protein that undergoes very little turnover, but sustained exposure to proteases may lead to irreversible and severe damage, and thus to functional loss of the elastic fiber network. Hence, it is a key issue to understand which enzymes actually initiate elastolysis under certain pathological conditions or during intrinsic aging. In this paper, we provide a complete workflow for isolation of pure and intact elastin from very small tissue samples to test enzymes for their elastolytic potential. This workflow was applied to skin samples from variously aged individuals, and it was found that strong differences exist in the degradability of the elastins investigated. In summary, human leukocyte elastase was unable to degrade intact elastin fibers but hydrolyzed elastin derived from the skin of old people. However, cathepsin G cleaved all elastin samples, even those derived from younger individuals. These results indicate that human leukocyte elastase is not a driving force for elastolysis, but may nevertheless promote further breakdown of elastic fibers after the action of other enzymes such as cathepsin G. © 2012 The Authors Journal compilation © 2012 FEBS.

  1. Facial lesions in piglets with intact or grinded teeth

    Directory of Open Access Journals (Sweden)

    Hansson Monica

    2012-04-01

    Full Text Available Abstract Background Piglets are born with eight sharp teeth that during nursing can cause facial lesions on littermates and teat lesions on the sow. Teeth grinding in piglets is therefore often practiced to reduce these lesions. The aim of this study was to assess the consequences of grinding piglet teeth in regard to the occurrence of lesions. In this study the piglets' teeth were grinded in 28 litters, and in 36 litters the piglets' teeth were kept intact. Twice, one time during the first week and one time during the second week after birth facial lesions of the piglets were scored and the teats of the sows were examined for lesions. The facial lesion score accounted for the amount and severity of lesions. The individual observations on piglets in the litter were synthesized in a litter facial lesion score. Findings 69.8% and 43.5% of the piglets had facial lesions in week 1 and week 2 respectively. The effect of treatment was not significant on litter facial lesion score. The litter facial lesion score was higher in week 1 than in week 2 (p p = 0.003 than in small litters. Mortality between week 1 and week 2 was higher in litters with intact teeth (p = 0.02. Sow teat lesions only occurred if litters had intact teeth. Conclusions According to our results teeth grinding is only justifiable in large litters.

  2. Modified Madigan Prostatectomy: A Procedure Preserved Prostatic Urethra Intact

    Institute of Scientific and Technical Information of China (English)

    LU Jun; YE Zhangqun; HU Weilie

    2005-01-01

    Summary: A total of 92 patients with benign prostatic hyperplasia (BPH) were subjected to modified Madigan prostatectomy (MPC) for a much satisfactory effect in open prostatectomy surgery. Exposing anterior prostatic urethra near the bladder neck and conjunct cystotomy modified the MPC procedure. This modified procedure preserved prostatic urethra intact and could also deal with intracystic lesions at the same time. The intact of prostatic urethra was kept completely or largely in 86 cases. The amount of blood loss during modified procedure was less. The mean operative time was 105 min. Seventy patients had been followed up for 3-24 months. The postoperative average Qmax was 19.2 ml/s. The cystourethrography revealed that the urethra and bladder neck were intact in 10 patients postoperatively. Furthermore, the prostatic urethra was obviously wider after modified MPC. The modified MPC can reduce the occurrence of urethra injury and enlarge the MPC indications. The modified technique is easy to perform with less complications and much satisfactory clinical result.

  3. High resolution X-ray structures of mouse major urinary protein nasal isoform in complex with pheromones

    Science.gov (United States)

    Perez-Miller, Samantha; Zou, Qin; Novotny, Milos V; Hurley, Thomas D

    2010-01-01

    In mice, the major urinary proteins (MUP) play a key role in pheromonal communication by binding and transporting semiochemicals. MUP-IV is the only isoform known to be expressed in the vomeronasal mucosa. In comparison with the MUP isoforms that are abundantly excreted in the urine, MUP-IV is highly specific for the male mouse pheromone 2-sec-butyl-4,5-dihydrothiazole (SBT). To examine the structural basis of this ligand preference, we determined the X-ray crystal structure of MUP-IV bound to three mouse pheromones: SBT, 2,5-dimethylpyrazine, and 2-heptanone. We also obtained the structure of MUP-IV with 2-ethylhexanol bound in the cavity. These four structures show that relative to the major excreted MUP isoforms, three amino acid substitutions within the binding calyx impact ligand coordination. The F103 for A along with F54 for L result in a smaller cavity, potentially creating a more closely packed environment for the ligand. The E118 for G substitution introduces a charged group into a hydrophobic environment. The sidechain of E118 is observed to hydrogen bond to polar groups on all four ligands with nearly the same geometry as seen for the water-mediated hydrogen bond network in the MUP-I and MUP-II crystal structures. These differences in cavity size and interactions between the protein and ligand are likely to contribute to the observed specificity of MUP-IV. PMID:20509168

  4. High resolution X-ray structures of mouse major urinary protein nasal isoform in complex with pheromones

    Energy Technology Data Exchange (ETDEWEB)

    Perez-Miller, Samantha; Zou, Qin; Novotny, Milos V.; Hurley, Thomas D. (Indiana-Med); (Indiana)

    2010-09-07

    In mice, the major urinary proteins (MUP) play a key role in pheromonal communication by binding and transporting semiochemicals. MUP-IV is the only isoform known to be expressed in the vomeronasal mucosa. In comparison with the MUP isoforms that are abundantly excreted in the urine, MUP-IV is highly specific for the male mouse pheromone 2-sec-butyl-4,5-dihydrothiazole (SBT). To examine the structural basis of this ligand preference, we determined the X-ray crystal structure of MUP-IV bound to three mouse pheromones: SBT, 2,5-dimethylpyrazine, and 2-heptanone. We also obtained the structure of MUP-IV with 2-ethylhexanol bound in the cavity. These four structures show that relative to the major excreted MUP isoforms, three amino acid substitutions within the binding calyx impact ligand coordination. The F103 for A along with F54 for L result in a smaller cavity, potentially creating a more closely packed environment for the ligand. The E118 for G substitution introduces a charged group into a hydrophobic environment. The sidechain of E118 is observed to hydrogen bond to polar groups on all four ligands with nearly the same geometry as seen for the water-mediated hydrogen bond network in the MUP-I and MUP-II crystal structures. These differences in cavity size and interactions between the protein and ligand are likely to contribute to the observed specificity of MUP-IV.

  5. High resolution X-ray structures of mouse major urinary protein nasal isoform in complex with pheromones.

    Science.gov (United States)

    Perez-Miller, Samantha; Zou, Qin; Novotny, Milos V; Hurley, Thomas D

    2010-08-01

    In mice, the major urinary proteins (MUP) play a key role in pheromonal communication by binding and transporting semiochemicals. MUP-IV is the only isoform known to be expressed in the vomeronasal mucosa. In comparison with the MUP isoforms that are abundantly excreted in the urine, MUP-IV is highly specific for the male mouse pheromone 2-sec-butyl-4,5-dihydrothiazole (SBT). To examine the structural basis of this ligand preference, we determined the X-ray crystal structure of MUP-IV bound to three mouse pheromones: SBT, 2,5-dimethylpyrazine, and 2-heptanone. We also obtained the structure of MUP-IV with 2-ethylhexanol bound in the cavity. These four structures show that relative to the major excreted MUP isoforms, three amino acid substitutions within the binding calyx impact ligand coordination. The F103 for A along with F54 for L result in a smaller cavity, potentially creating a more closely packed environment for the ligand. The E118 for G substitution introduces a charged group into a hydrophobic environment. The sidechain of E118 is observed to hydrogen bond to polar groups on all four ligands with nearly the same geometry as seen for the water-mediated hydrogen bond network in the MUP-I and MUP-II crystal structures. These differences in cavity size and interactions between the protein and ligand are likely to contribute to the observed specificity of MUP-IV.

  6. Computer mouse movement patterns: A potential marker of mild cognitive impairment

    OpenAIRE

    Seelye, Adriana; Hagler, Stuart; Mattek, Nora; Diane B Howieson; Wild, Katherine; Dodge, Hiroko H.; Kaye, Jeffrey A.

    2015-01-01

    Introduction Subtle changes in cognitively demanding activities occur in mild cognitive impairment (MCI) but are difficult to assess with conventional methods. In an exploratory study, we examined whether patterns of computer mouse movements obtained from routine home computer use discriminated between older adults with and without MCI. Methods Participants were 42 cognitively intact and 20 older adults with MCI enrolled in a longitudinal study of in-home monitoring technologies. Mouse pointe...

  7. NCBI nr-aa BLAST: CBRC-OCUN-01-1001 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OCUN-01-1001 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasa

  8. NCBI nr-aa BLAST: CBRC-PABE-04-0003 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PABE-04-0003 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasa

  9. NCBI nr-aa BLAST: CBRC-MMUS-06-0138 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0138 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasa

  10. NCBI nr-aa BLAST: CBRC-STRI-01-2495 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-2495 ref|NP_444448.1| vomeronasal 1 receptor, A3 [Mus musculus] sp|Q9E...Q52|VN1A3_MOUSE RecName: Full=Vomeronasal type-1 receptor A3; AltName: Full=Vomeronasal type-1 receptor A6 g...b|AAG42076.1|AF291482_1 vomeronasal receptor V1RA3 [Mus musculus] dbj|BAB79212.1| vomeronasal receptor 1 A6 ...[Mus musculus] gb|AAI41612.1| Vomeronasal 1 receptor, A3 [synthetic construct] gb|AAI40226.1| Vomeronasa

  11. NCBI nr-aa BLAST: CBRC-STRI-01-2627 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-2627 sp|Q8VBS7|VN1A6_MOUSE RecName: Full=Vomeronasal type-1 receptor A...6; AltName: Full=Vomeronasal type-1 receptor A3; AltName: Full=Vomeronasal type-1 receptor A1; AltName: Full=Vomeronasa...l type-1 receptor A10 dbj|BAB79209.1| vomeronasal receptor 1 A1 [Mus musculus] dbj|BAB79210.1| vomeronasa...l receptor 1 A3 [Mus musculus] gb|AAK98772.1| vomeronasal receptor V1RA10

  12. NCBI nr-aa BLAST: CBRC-CFAM-20-0001 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CFAM-20-0001 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasa

  13. NCBI nr-aa BLAST: CBRC-RNOR-04-0258 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-04-0258 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasa

  14. NCBI nr-aa BLAST: CBRC-STRI-01-0514 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-0514 ref|NP_444458.1| vomeronasal 1 receptor, B7 [Mus musculus] sp|Q9E...Q46|VN1B7_MOUSE RecName: Full=Vomeronasal type-1 receptor B7; AltName: Full=Vomeronasal type-1 receptor A11 ...gb|AAG42087.1|AF291493_1 vomeronasal receptor V1RB7 [Mus musculus] dbj|BAB79217.1| vomeronasal receptor 1 A1...1 [Mus musculus] gb|AAI41627.1| Vomeronasal 1 receptor, B7 [synthetic construct] gb|AAI40251.1| Vomeronasa

  15. NCBI nr-aa BLAST: CBRC-RNOR-04-0245 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-04-0245 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasa

  16. NCBI nr-aa BLAST: CBRC-STRI-01-2627 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-2627 ref|NP_444448.1| vomeronasal 1 receptor, A3 [Mus musculus] sp|Q9E...Q52|VN1A3_MOUSE RecName: Full=Vomeronasal type-1 receptor A3; AltName: Full=Vomeronasal type-1 receptor A6 g...b|AAG42076.1|AF291482_1 vomeronasal receptor V1RA3 [Mus musculus] dbj|BAB79212.1| vomeronasal receptor 1 A6 ...[Mus musculus] gb|AAI41612.1| Vomeronasal 1 receptor, A3 [synthetic construct] gb|AAI40226.1| Vomeronasa

  17. NCBI nr-aa BLAST: CBRC-STRI-01-2796 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-2796 ref|NP_444458.1| vomeronasal 1 receptor, B7 [Mus musculus] sp|Q9E...Q46|VN1B7_MOUSE RecName: Full=Vomeronasal type-1 receptor B7; AltName: Full=Vomeronasal type-1 receptor A11 ...gb|AAG42087.1|AF291493_1 vomeronasal receptor V1RB7 [Mus musculus] dbj|BAB79217.1| vomeronasal receptor 1 A1...1 [Mus musculus] gb|AAI41627.1| Vomeronasal 1 receptor, B7 [synthetic construct] gb|AAI40251.1| Vomeronasa

  18. NCBI nr-aa BLAST: CBRC-CPOR-01-0582 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CPOR-01-0582 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasa

  19. NCBI nr-aa BLAST: CBRC-STRI-01-2495 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-2495 sp|Q8VBS7|VN1A6_MOUSE RecName: Full=Vomeronasal type-1 receptor A...6; AltName: Full=Vomeronasal type-1 receptor A3; AltName: Full=Vomeronasal type-1 receptor A1; AltName: Full=Vomeronasa...l type-1 receptor A10 dbj|BAB79209.1| vomeronasal receptor 1 A1 [Mus musculus] dbj|BAB79210.1| vomeronasa...l receptor 1 A3 [Mus musculus] gb|AAK98772.1| vomeronasal receptor V1RA10

  20. NCBI nr-aa BLAST: CBRC-STRI-01-2691 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-2691 sp|Q8VBS7|VN1A6_MOUSE RecName: Full=Vomeronasal type-1 receptor A...6; AltName: Full=Vomeronasal type-1 receptor A3; AltName: Full=Vomeronasal type-1 receptor A1; AltName: Full=Vomeronasa...l type-1 receptor A10 dbj|BAB79209.1| vomeronasal receptor 1 A1 [Mus musculus] dbj|BAB79210.1| vomeronasa...l receptor 1 A3 [Mus musculus] gb|AAK98772.1| vomeronasal receptor V1RA10

  1. NCBI nr-aa BLAST: CBRC-MMUS-06-0126 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0126 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasa

  2. NCBI nr-aa BLAST: CBRC-RNOR-04-0242 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-04-0242 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasa

  3. NCBI nr-aa BLAST: CBRC-OCUN-01-0501 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OCUN-01-0501 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8VIC7|VN1A2_MOUSE Vomeronas...al type-1 receptor A2 (Pheromone receptor 2) (Vomeronasal type-1 receptor A9) (mV1R...2) dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal 1 receptor, A2 [synth...etic construct] gb|AAI40265.1| Vomeronasal 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasa

  4. Micron-scale voltage and [Ca2+]i imaging in the intact heart

    Directory of Open Access Journals (Sweden)

    Xiao-long eLu

    2014-12-01

    Full Text Available Studies in isolated cardiomyocytes have provided tremendous information at the cellular and molecular level concerning regulation of transmembrane voltage (Vm and intracellular calcium ([Ca2+]i. The ability to use the information gleaned to gain insight into the function of ion channels and Ca2+ handling proteins in a more complex system, e. g., the intact heart, has remained a challenge. We have developed laser scanning fluorescence microscopy-based approaches to monitor, at the sub-cellular to multi-cellular level in the immobilized, Langendorff-perfused mouse heart, dynamic changes in [Ca2+]i and Vm. This article will review the use of single- or dual-photon laser scanning microscopy [Ca2+]i imaging in conjunction with transgenic reporter technology to a interrogate the extent to which transplanted, donor-derived myocytes or cardiac stem cell-derived de novo myocytes are capable of forming a functional syncytium with the pre-existing myocardium, using entrainment of [Ca2+]i transients by the electrical activity of the recipient heart as a surrogate for electrical coupling, and b characterize the Ca2+ handling phenotypes of cellular implants. Further, we will review the ability of laser scanning fluorescence microscopy in conjunction with a fast-response voltage-sensitive to resolve, on a subcellular level in Langendorff-perfused mouse hearts, Vm dynamics that typically occur during the course of a cardiac action potential. Specifically, the utility of this technique to measure microscopic-scale voltage gradients in the normal and diseased heart is discussed.

  5. Sphingolipid metabolism in organotypic mouse keratinocyte cultures

    Energy Technology Data Exchange (ETDEWEB)

    Madison, K.C.; Swartzendruber, D.C.; Wertz, P.W.; Downing, D.T. (Univ. of Iowa College of Medicine, Iowa City (USA))

    1990-12-01

    Ceramides are the dominant component of the stratum corneum intercellular lipid lamellae, which constitute the epidermal permeability barrier. Only pig and human epidermal ceramides have been extensively characterized and the structures of the ceramides of cultured keratinocytes have not been previously investigated. In the present studies, we have characterized the ceramides synthesized by organotypic lifted mouse keratinocyte cultures for the first time and compared them to the ceramides of intact mouse epidermis. Both mouse epidermis and cultures contained five ceramides, ceramide 1 being the least polar and ceramide 5 the most polar. Ceramide 1 was a group of acylceramides, i.e., very-long-chain omega-hydroxyceramides with an ester-linked nonhydroxy fatty acid. Ceramide 2 contained medium-length saturated nonhydroxy fatty acids. (In culture, the ceramide 2 band was split into two parts with the slightly more polar ceramide 2' containing short-chain saturated nonhydroxy fatty acids.) Ceramide 5 contained short-chain alpha-hydroxy fatty acids. The structures of ceramides 1, 2, and 5 were analagous to those of pig and human epidermis. Mouse epidermal ceramide 3 was quite unusual, containing beta-hydroxy fatty acids, a structure not previously identified among mammalian ceramides. In contrast, culture ceramide 3 was composed of omega-hydroxy fatty acids with a chain-length distribution similar to that of ceramide 1. Mouse ceramide 4 was composed of fatty acids with chromatographic mobility similar to hydroxy fatty acids but with different chemical reactivity; it remains only partially characterized. Culture ceramide 4 was present in quantities too small for analysis. All ceramides in mouse epidermis and cultures contained only sphingosine bases, whereas pig and human ceramides also contain phytosphingosine.

  6. Intact Endogenous Metabolite Analysis of Mice Liver by Probe Electrospray Ionization/Triple Quadrupole Tandem Mass Spectrometry and Its Preliminary Application to in Vivo Real-Time Analysis.

    Science.gov (United States)

    Zaitsu, Kei; Hayashi, Yumi; Murata, Tasuku; Ohara, Tomomi; Nakagiri, Kenta; Kusano, Maiko; Nakajima, Hiroki; Nakajima, Tamie; Ishikawa, Tetsuya; Tsuchihashi, Hitoshi; Ishii, Akira

    2016-04-05

    Probe electrospray ionization (PESI) is a recently developed ionization technique that enables the direct detection of endogenous compounds like metabolites without sample preparation. In this study, we have demonstrated the first combination use of PESI with triple quadrupole tandem mass spectrometry (MS/MS), which was then applied to intact endogenous metabolite analysis of mice liver, achieving detection of 26 metabolites including amino acids, organic acids, and sugars. To investigate its practicality, metabolic profiles of control and CCl4-induced acute hepatic injury mouse model were measured by the developed method. Results showed clear separation of the two groups in score plots of principal component analysis and identified taurine as the primary contributor to group separation. The results were further validated by the established gas chromatography/MS/MS method, demonstrating the present method's usefulness. In addition, we preliminarily applied the method to real-time analysis of an intact liver of a living mouse. We successfully achieved monitoring of the real-time changes of two tricarboxylic acid cycle intermediates, α-ketoglutaric acid and fumaric acid, in the liver immediately after pyruvic acid injection via a cannulated tube to the portal vein. The present method achieved an intact analysis of metabolites in liver without sample preparation, and it also demonstrates future possibility to establish in vivo real-time metabolome analysis of living animals by PESI/MS/MS.

  7. Feasible pickup from intact ossicular chain with floating piezoelectric microphone

    Directory of Open Access Journals (Sweden)

    Kang Hou-Yong

    2012-02-01

    Full Text Available Abstract Objectives Many microphones have been developed to meet with the implantable requirement of totally implantable cochlear implant (TICI. However, a biocompatible one without destroying the intactness of the ossicular chain still remains under investigation. Such an implantable floating piezoelectric microphone (FPM has been manufactured and shows an efficient electroacoustic performance in vitro test at our lab. We examined whether it pick up sensitively from the intact ossicular chain and postulated whether it be an optimal implantable one. Methods Animal controlled experiment: five adult cats (eight ears were sacrificed as the model to test the electroacoustic performance of the FPM. Three groups were studied: (1 the experiment group (on malleus: the FPM glued onto the handle of the malleus of the intact ossicular chains; (2 negative control group (in vivo: the FPM only hung into the tympanic cavity; (3 positive control group (Hy-M30: a HiFi commercial microphone placed close to the site of the experiment ear. The testing speaker played pure tones orderly ranged from 0.25 to 8.0 kHz. The FPM inside the ear and the HiFi microphone simultaneously picked up acoustic vibration which recorded as .wav files to analyze. Results The FPM transformed acoustic vibration sensitively and flatly as did the in vitro test across the frequencies above 2.0 kHz, whereas inefficiently below 1.0 kHz for its overloading mass. Although the HiFi microphone presented more efficiently than the FPM did, there was no significant difference at 3.0 kHz and 8.0 kHz. Conclusions It is feasible to develop such an implantable FPM for future TICIs and TIHAs system on condition that the improvement of Micro Electromechanical System and piezoelectric ceramic material technology would be applied to reduce its weight and minimize its size.

  8. A case of mixed transcortical aphasia with intact naming.

    Science.gov (United States)

    Heilman, K M; Tucker, D M; Valenstein, E

    1976-09-01

    Altholgh Lichtheim recognized that Wernicke's 'reflex arch' (primary auditory area, to Wernicke's area, to Broca's area, to primary motor area) was important for repetition, he recognized that other areas of the brain (for example, area of concepts or semantic area) must be important in comprehension and voluntary speech. He suggested that Wernicke's area (phonemic area) not only projected to Broca's area (as Wernicke suggested) but that it also projected to the area of concepts. A lesion of this latter pathway or in the area of concepts would produce a syndrome where repetition was intact but comprehension was impaired (e.g. transcortical sensory aphasia). Lichtheim also thought that the area of concepts projected directly to Broca's area and that voluntary speech was mediated by this pathway. Although Lichtheim's model could explain the mechanism underlying transcortical aphasia, his schema could not explain anomic aphasia. Unlike Lichtheim's schema, Kussmaul's schema suggested that the area of concepts projects back to Wernicke's area before projecting to Broca's area. With this schema, a patient with a hypothetical lesion which interrupted the pathway from the area of concepts to Wernicke's area (but did not interrupt the pathway from Wernicke's area to the area of concepts) should be anomic, with normal comprehension and repetition. In order for this latter schema to be plausible there should also be a lesion which interrupts the pathway from Wernicke's area to the area of concepts but does not interrupt the pathway which goes from the area of concepts to Wernicke's area. A patient with this hypothetical lesion should comprehend poorly; however, in spite of poor comprehension, naming and repetition should be intact. We report a patient who demonstrates poor comprehension with intact naming and repetition. This patient could also read aloud but could not comprehend written language. Not only could this patient name objects but he could demonstrate their use

  9. Direct detection of radicals in intact soybean nodules

    DEFF Research Database (Denmark)

    Mathieu, C; Moreau, S; Frendo, P

    1998-01-01

    Electron paramagnetic resonance spectroscopy has been employed to examine the nature of the metal ions and radicals present in intact root nodules of soybean plants grown in the absence of nitrate. The spectra obtained from nodules of different ages using this non-invasive technique show dramatic...... of the soybean plants, in a manner analogous to that recently described for Lupinus albus. This Lb-NO complex is present at lower concentrations in older nodules, and is almost completely absent from senescent nodules. Exposure of young and mature nodules to oxidant stress, in the form of hydrogen peroxide...

  10. Rhesus monkey brain imaging through intact skull with thermoacoustic tomography

    OpenAIRE

    Xu, Yuan; Wang, Lihong V.

    2006-01-01

    Two-dimensional microwave-induced thermoacoustic tomography (TAT) is applied to imaging the Rhesus monkey brain through the intact skull. To reduce the wavefront distortion caused by the skull, only the low-frequency components of the thermoacoustic signals (< 1 MHz) are used to reconstruct the TAT images. The methods of signal processing and image reconstruction are validated by imaging a lamb kidney. The resolution of the system is found to be 4 mm when we image a 1-month-old monkey head co...

  11. Fast imaging of intact and shattered cryogenic neon pellets

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Zhehui, E-mail: zwang@lanl.gov [Los Alamos National Laboratory, Los Alamos, New Mexico 87545 (United States); Combs, S. K.; Baylor, L. R.; Foust, C. R.; Lyttle, M. S.; Meitner, S. J.; Rasmussen, D. A. [Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831 (United States)

    2014-11-15

    Compact condensed-matter injection technologies are increasingly used in magnetic fusion. One recent application is in disruption mitigation. An imaging system with less-than-100-µm- and sub-µs-resolution is described and used to characterize intact and shattered cryogenic neon pellets. Shattered pellets contain fine particles ranging from tens of µm to about 7 mm. Time-of-flight analyses indicate that pellets could slow down if hitting the wall of the guide tube. Fast high-resolution imaging systems are thus useful to neon and other condensed-matter injector development.

  12. Fast Imaging of Intact and Shattered Cryogenic Neon Pellets

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Zhehui [Los Alamos National Laboratory (LANL); Combs, Stephen Kirk [ORNL; Baylor, Larry R [ORNL; Foust, Charles R [ORNL; Lyttle, Mark S [ORNL; Meitner, Steven J [ORNL; Rasmussen, David A [ORNL

    2014-01-01

    Compact condensed-matter injection technologies are increasingly used in magnetic fusion. One recent application is in disruption mitigation. An imaging system with less-than-100- m- and sub- s-resolution is described and used to characterize intact and shattered cryogenic neon pellets. Shattered pellets contain fine particles ranging from tens of m to about 7 mm. Time-of-flight analyses indicate that pellets could slow down if hitting the wall of the guide tube. Fast high-resolution imaging systems are thus useful to neon and other condensed-matter injector development.

  13. Fast imaging of intact and shattered cryogenic neon pellets.

    Science.gov (United States)

    Wang, Zhehui; Combs, S K; Baylor, L R; Foust, C R; Lyttle, M S; Meitner, S J; Rasmussen, D A

    2014-11-01

    Compact condensed-matter injection technologies are increasingly used in magnetic fusion. One recent application is in disruption mitigation. An imaging system with less-than-100-µm- and sub-µs-resolution is described and used to characterize intact and shattered cryogenic neon pellets. Shattered pellets contain fine particles ranging from tens of µm to about 7 mm. Time-of-flight analyses indicate that pellets could slow down if hitting the wall of the guide tube. Fast high-resolution imaging systems are thus useful to neon and other condensed-matter injector development.

  14. A radiological study on lumbar spondylolisthesis with intact neural arch

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Myung Ju; Suh, Young Jun; Joe, Seong Jeh; Yoon, Jong Sup [Han-Gang Sung-Shim Hospital, Seoul (Korea, Republic of)

    1980-12-15

    The purpose of this paper is to evaluate and analyze the radiological findings of 13 cases of lumbar spondylolisthesis with intact neural arch at Han-Gang Sung- Shim Hospital during period from Jan. 1975 to June 1980. 1. The age distribution of the patients varied from 31 to 83 years old. The average of the age was 53 years old. 12 patients were female and only one patient was male. 2. All cases revealed anterior slipping of the L4 body on the L5 body, of which degree was classified by Meyerding's method, first degree in 12 patients and second degree in one patient. 3. The angle between the pedicle and inferior articular facet of affected vertebra was measured by Junghann's method. The average of angle was 123 degree. 4. Of 13 cases, 2 cases who had neural compression caused by dislocation of the vertebral bodies and intrusion of lamina and arthritic facets into stenotic spinal canal treated by laminar decompression including foraminotomy. Good results were obtained. 5. The secondary degenerative change of the vertebra was considered to be the cause of the spondylolisthesis with intact neural arch.

  15. Selective ion changes during spontaneous mitochondrial transients in intact astrocytes.

    Directory of Open Access Journals (Sweden)

    Guillaume Azarias

    Full Text Available The bioenergetic status of cells is tightly regulated by the activity of cytosolic enzymes and mitochondrial ATP production. To adapt their metabolism to cellular energy needs, mitochondria have been shown to exhibit changes in their ionic composition as the result of changes in cytosolic ion concentrations. Individual mitochondria also exhibit spontaneous changes in their electrical potential without altering those of neighboring mitochondria. We recently reported that individual mitochondria of intact astrocytes exhibit spontaneous transient increases in their Na(+ concentration. Here, we investigated whether the concentration of other ionic species were involved during mitochondrial transients. By combining fluorescence imaging methods, we performed a multiparameter study of spontaneous mitochondrial transients in intact resting astrocytes. We show that mitochondria exhibit coincident changes in their Na(+ concentration, electrical potential, matrix pH and mitochondrial reactive oxygen species production during a mitochondrial transient without involving detectable changes in their Ca(2+ concentration. Using widefield and total internal reflection fluorescence imaging, we found evidence for localized transient decreases in the free Mg(2+ concentration accompanying mitochondrial Na(+ spikes that could indicate an associated local and transient enrichment in the ATP concentration. Therefore, we propose a sequential model for mitochondrial transients involving a localized ATP microdomain that triggers a Na(+-mediated mitochondrial depolarization, transiently enhancing the activity of the mitochondrial respiratory chain. Our work provides a model describing ionic changes that could support a bidirectional cytosol-to-mitochondria ionic communication.

  16. Selective ion changes during spontaneous mitochondrial transients in intact astrocytes.

    Science.gov (United States)

    Azarias, Guillaume; Chatton, Jean-Yves

    2011-01-01

    The bioenergetic status of cells is tightly regulated by the activity of cytosolic enzymes and mitochondrial ATP production. To adapt their metabolism to cellular energy needs, mitochondria have been shown to exhibit changes in their ionic composition as the result of changes in cytosolic ion concentrations. Individual mitochondria also exhibit spontaneous changes in their electrical potential without altering those of neighboring mitochondria. We recently reported that individual mitochondria of intact astrocytes exhibit spontaneous transient increases in their Na(+) concentration. Here, we investigated whether the concentration of other ionic species were involved during mitochondrial transients. By combining fluorescence imaging methods, we performed a multiparameter study of spontaneous mitochondrial transients in intact resting astrocytes. We show that mitochondria exhibit coincident changes in their Na(+) concentration, electrical potential, matrix pH and mitochondrial reactive oxygen species production during a mitochondrial transient without involving detectable changes in their Ca(2+) concentration. Using widefield and total internal reflection fluorescence imaging, we found evidence for localized transient decreases in the free Mg(2+) concentration accompanying mitochondrial Na(+) spikes that could indicate an associated local and transient enrichment in the ATP concentration. Therefore, we propose a sequential model for mitochondrial transients involving a localized ATP microdomain that triggers a Na(+)-mediated mitochondrial depolarization, transiently enhancing the activity of the mitochondrial respiratory chain. Our work provides a model describing ionic changes that could support a bidirectional cytosol-to-mitochondria ionic communication.

  17. Large scale expression and purification of mouse melanopsin-L in the baculovirus expression system

    NARCIS (Netherlands)

    Shirzad-Wasei, N.; Oostrum, J. van; Bovee-Geurts, P.H.M.; Wasserman, M.; Bosman, G.J.C.G.M.; Degrip, W.J.

    2013-01-01

    Melanopsin is the mammalian photopigment that primarily mediates non-visual photoregulated physiology. So far, this photopigment is poorly characterized with respect to structure and function. Here, we report large-scale production and purification of the intact long isoform of mouse melanopsin (mel

  18. NCBI nr-aa BLAST: CBRC-MMUS-20-0071 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-20-0071 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 1e-137 79% ...

  19. NCBI nr-aa BLAST: CBRC-RNOR-04-0256 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-04-0256 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 2e-74 71% ...

  20. NCBI nr-aa BLAST: CBRC-MMUS-06-0128 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0128 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 1e-175 99% ...

  1. NCBI nr-aa BLAST: CBRC-RNOR-04-0239 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-04-0239 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 1e-139 80% ...

  2. NCBI nr-aa BLAST: CBRC-RNOR-04-0249 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-04-0249 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 1e-133 77% ...

  3. NCBI nr-aa BLAST: CBRC-RNOR-04-0259 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-04-0259 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 1e-124 73% ...

  4. NCBI nr-aa BLAST: CBRC-MMUS-06-0123 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0123 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 1e-137 79% ...

  5. NCBI nr-aa BLAST: CBRC-OCUN-01-0056 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OCUN-01-0056 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 8e-47 57% ...

  6. NCBI nr-aa BLAST: CBRC-MMUS-06-0141 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0141 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 1e-141 82% ...

  7. NCBI nr-aa BLAST: CBRC-MMUS-06-0122 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0122 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 1e-142 80% ...

  8. NCBI nr-aa BLAST: CBRC-MMUS-06-0131 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0131 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 1e-137 80% ...

  9. NCBI nr-aa BLAST: CBRC-MMUS-06-0136 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0136 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 1e-138 77% ...

  10. NCBI nr-aa BLAST: CBRC-RNOR-04-0238 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-04-0238 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 1e-135 82% ...

  11. NCBI nr-aa BLAST: CBRC-RNOR-04-0255 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-04-0255 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 1e-122 74% ...

  12. NCBI nr-aa BLAST: CBRC-MMUS-06-0135 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0135 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 1e-141 82% ...

  13. NCBI nr-aa BLAST: CBRC-CPOR-01-0246 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CPOR-01-0246 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 4e-32 55% ...

  14. NCBI nr-aa BLAST: CBRC-OCUN-01-0645 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OCUN-01-0645 sp|Q9EQ47|VN1B4_MOUSE Vomeronasal type-1 receptor B4 (Vomeronasal... type-1 receptor A10) (Vomeronasal type-1 receptor B11) gb|AAI07186.1| V1rb4 protein [Mus musculus] Q9EQ47 4e-49 57% ...

  15. Holographic intravital microscopy for 2-D and 3-D imaging intact circulating blood cells in microcapillaries of live mice

    CERN Document Server

    Kim, Kyoohyun; Park, Inwon; Kim, Pilhan; Park, YongKeun

    2016-01-01

    Intravital microscopy is an essential tool that reveals behaviours of live cells under conditions close to natural physiological states. So far, although various approaches for imaging cells in vivo have been proposed, most require the use of labelling and also provide only qualitative imaging information. Holographic imaging approach based on measuring the refractive index distributions of cells, however, circumvent these problems and offer quantitative and label-free imaging capability. Here, we demonstrate in vivo two- and three-dimensional holographic imaging of circulating blood cells in intact microcapillaries of live mice. The measured refractive index distributions of blood cells provide morphological and biochemical properties including three-dimensional cell shape, haemoglobin concentration, and haemoglobin contents at the individual cell level. With the present method, alterations in blood flow dynamics in live healthy and sepsis-model mouse were also investigated.

  16. Modification of bacterial artificial chromosomes (BACs) and preparation of intact BAC DNA for generation of transgenic mice.

    Science.gov (United States)

    Gong, Shiaoching; Yang, X William

    2005-05-01

    BAC transgenesis is a powerful tool for the study of gene expression and gene function in the mouse in vivo. In this unit, detailed protocols are provided for modification (i.e., marker gene insertion, deletion, or point mutation) of BACs by homologous recombination in E. coli. This method utilizes a shuttle vector that allows transient expression of the E. coli RecA gene to support homologous recombination in the BAC host bacteria. In addition, two protocols are provided for purification of BAC DNA for microinjection to generate transgenic mice. Since BAC DNA is prone to degradation, which may introduce positional effects in transgenic mice, two methods are given for purification of intact BAC DNA for subsequent microinjection.

  17. Gaze beats mouse

    DEFF Research Database (Denmark)

    Mateo, Julio C.; San Agustin, Javier; Hansen, John Paulin

    2008-01-01

    Facial EMG for selection is fast, easy and, combined with gaze pointing, it can provide completely hands-free interaction. In this pilot study, 5 participants performed a simple point-and-select task using mouse or gaze for pointing and a mouse button or a facial-EMG switch for selection. Gaze...... pointing was faster than mouse pointing, while maintaining a similar error rate. EMG and mouse-button selection had a comparable performance. From analyses of completion time, throughput and error rates, we concluded that the combination of gaze and facial EMG holds potential for outperforming the mouse....

  18. Diffusion tensor imaging measures of white matter compared to myelin basic protein immunofluorescence in tissue cleared intact brains

    Directory of Open Access Journals (Sweden)

    Eric H. Chang

    2017-02-01

    Full Text Available We provide datasets from combined ex vivo diffusion tensor imaging (DTI and Clear Lipid-exchanged, Anatomically Rigid, Imaging/immunostaining compatible, Tissue hYdrogel (CLARITY performed on intact mouse brains. DTI-derived measures of fractional anisotropy (FA, radial diffusivity (RD, and axial diffusivity (AD were compared to antibody-based labeling of myelin basic protein (MBP, as measured by fluorescence microscopy. We used a customized CLARITY hydrogel solution to facilitate whole brain tissue clearing and subsequent immunolabeling. We describe how CLARITY was made compatible with magnetic resonance imaging with the intention of facilitating future multimodal imaging studies that may combine noninvasive imaging with 3D immunohistochemistry. These data and methods are related to the accompanying research article entitled, ‘The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains’ (E.H. Chang, M. Argyelan, M. Aggarwal, T-S. Chandon, K.H. Karlsgodt, S. Mori, A.K. Malhotra, 2016 [1].

  19. Cell types, circuits, and receptive fields in the mouse visual cortex.

    Science.gov (United States)

    Niell, Cristopher M

    2015-07-08

    Over the past decade, the mouse has emerged as an important model system for studying cortical function, owing to the advent of powerful tools that can record and manipulate neural activity in intact neural circuits. This advance has been particularly prominent in the visual cortex, where studies in the mouse have begun to bridge the gap between cortical structure and function, allowing investigators to determine the circuits that underlie specific visual computations. This review describes the advances in our understanding of the mouse visual cortex, including neural coding, the role of different cell types, and links between vision and behavior, and discusses how recent findings and new approaches can guide future studies.

  20. Calcium Activation Profile In Electrically Stimulated Intact Rat Heart Cells

    Science.gov (United States)

    Geerts, Hugo; Nuydens, Rony; Ver Donck, Luc; Nuyens, Roger; De Brabander, Marc; Borgers, Marcel

    1988-06-01

    Recent advances in fluorescent probe technology and image processing equipment have made available the measurement of calcium in living systems on a real-time basis. We present the use of the calcium indicator Fura-2 in intact normally stimulated rat heart cells for the spatial and dynamic measurement of the calcium excitation profile. After electric stimulation (1 Hz), the activation proceeds from the center of the myocyte toward the periphery. Within two frame times (80 ms), the whole cell is activated. The activation is slightly faster in the center of the cell than in the periphery. The mean recovery time is 200-400 ms. There is no difference along the cell's long axis. The effect of a beta-agonist and of a calcium antagonist is described.

  1. Complete posterior migration of intact vertebral body in spinal tuberculosis

    Directory of Open Access Journals (Sweden)

    Sridhar Krishnamurthy

    2009-01-01

    Full Text Available Spinal tuberculosis most commonly presents as a paradiscal lesion involving the disc space and adjacent vertebral bodies. Atypical forms of spinal tuberculosis have been described and are most often a result of posterior element involvement. The authors report a patient, who presented with complete posterior migration of an intact vertebral body, a complication of spinal tuberculosis that has not been reported till date. A 12-year-old girl with history of pulmonary tuberculosis presented with progressive paraparesis and back pain. Plain X-rays and MRI revealed that the L2 vertebral body had migrated posteriorly into the spinal canal, without significant movement of the posterior elements. The vertebral body was normal, with no erosion or bone loss. However, bilateral pedicle and facet joint involvement was seen. The neural elements were decompressed through an anterolateral retroperitoneal approach and the spine reconstructed. The authors present this rare manifestation of spinal tuberculosis and discuss the possible mechanisms of this presentation.

  2. In vitro adherence of Moraxella bovis to intact corneal epithelium.

    Science.gov (United States)

    Jackman, S H; Rosenbusch, R F

    1984-09-01

    An in vitro assay is described using radiolabeled Moraxella bovis for studying adherence to intact bovine corneal epithelial surfaces. The assay was optimized for time (45 min) and for the ratio of epithelial cells to bacteria (1:1000) that demonstrated a significant difference in adherence between M. bovis strain 118F, a piliated organism and a nonpiliated variant, strain 118F/4-2. Adherence of these organisms correlated with previous pathogenicity studies involving experimental infection of calves. Scanning electron microscopy of tissues treated in the assay revealed a predilection of M. bovis for dark epithelial cells and for association with depressions in the tissue surface. This assay technique is discussed in comparison with other in vitro adherence assay methods.

  3. Rhesus monkey brain imaging through intact skull with thermoacoustic tomography.

    Science.gov (United States)

    Xu, Yuan; Wang, Lihong V

    2006-03-01

    Two-dimensional microwave-induced thermoacoustic tomography (TAT) is applied to imaging the Rhesus monkey brain through the intact skull. To reduce the wavefront distortion caused by the skull, only the low-frequency components of the thermoacoustic signals (images. The methods of signal processing and image reconstruction are validated by imaging a lamb kidney. The resolution of the system is found to be 4 mm when we image a 1-month-old monkey head containing inserted needles. We also image the coronal and axial sections of a 7-month-old monkey head. Brain features that are 3 cm deep in the head are imaged clearly. Our results demonstrate that TAT has potential for use in portable, cost-effective imagers for pediatric brains.

  4. Label-free volumetric optical imaging of intact murine brains

    Science.gov (United States)

    Ren, Jian; Choi, Heejin; Chung, Kwanghun; Bouma, Brett E.

    2017-04-01

    A central effort of today’s neuroscience is to study the brain’s ’wiring diagram’. The nervous system is believed to be a network of neurons interacting with each other through synaptic connection between axons and dendrites, therefore the neuronal connectivity map not only depicts the underlying anatomy, but also has important behavioral implications. Different approaches have been utilized to decipher neuronal circuits, including electron microscopy (EM) and light microscopy (LM). However, these approaches typically demand extensive sectioning and reconstruction for a brain sample. Recently, tissue clearing methods have enabled the investigation of a fully assembled biological system with greatly improved light penetration. Yet, most of these implementations, still require either genetic or exogenous contrast labeling for light microscopy. Here we demonstrate a high-speed approach, termed as Clearing Assisted Scattering Tomography (CAST), where intact brains can be imaged at optical resolution without labeling by leveraging tissue clearing and the scattering contrast of optical frequency domain imaging (OFDI).

  5. Chemical Probes for Visualizing Intact Animal and Human Brain Tissue.

    Science.gov (United States)

    Lai, Hei Ming; Ng, Wai-Lung; Gentleman, Steve M; Wu, Wutian

    2017-06-22

    Newly developed tissue clearing techniques can be used to render intact tissues transparent. When combined with fluorescent labeling technologies and optical sectioning microscopy, this allows visualization of fine structure in three dimensions. Gene-transfection techniques have proved very useful in visualizing cellular structures in animal models, but they are not applicable to human brain tissue. Here, we discuss the characteristics of an ideal chemical fluorescent probe for use in brain and other cleared tissues, and offer a comprehensive overview of currently available chemical probes. We describe their working principles and compare their performance with the goal of simplifying probe selection for neuropathologists and stimulating probe development by chemists. We propose several approaches for the development of innovative chemical labeling methods which, when combined with tissue clearing, have the potential to revolutionize how we study the structure and function of the human brain. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. True navigation in migrating gulls requires intact olfactory nerves

    DEFF Research Database (Denmark)

    Wikelski, Martin; Arriero, Elena; Gagliardo, Anna;

    2015-01-01

    debated. In this experiment we subjected adult lesser black-backed gulls migrating from their Finnish/Russian breeding grounds (from >60°N) to Africa (to systems required for navigation. We translocated birds westward (1080 km) or eastward (885 km......) to simulate natural navigational challenges. When translocated westwards and outside their migratory corridor birds with olfactory nerve section kept a clear directional preference (southerly) but were unable to compensate for the displacement, while intact birds and gulls with the ophthalmic branch...... of the trigeminal nerve sectioned oriented towards their population-specific migratory corridor. Thus, air-borne olfactory information seems to be important for migrating gulls to navigate successfully in some circumstances....

  7. Synthesis of pennogenin utilizing the intact skeleton of diosgenin

    Institute of Scientific and Technical Information of China (English)

    TIAN; Weisheng; XU; Qihai; CHEN; Ling; ZHAO; Chunfeng

    2004-01-01

    The first synthesis of pennogenin, an aglycone of bioactive components of Chinese traditional medicine named "Chonglou"(Paris), starting from diosgenin, has been reported, which displays a new strategy of utilizing the resource compounds. According to this new strategy, the full and rational utilization of the intact skeleton and functional groups of starting material has been realized in the conversion of diosgenin to pennogenin. The key step for synthesis of pennogenin is the regioselective transformation of cholest-5-en-16,22-dion-3,26-diol to cholest5,16-dien-22-on-3,26-diol, which can be used to synthesize other bioactive steroids such as cephalostatin and OSW-1.

  8. Simple Genome Editing of Rodent Intact Embryos by Electroporation.

    Directory of Open Access Journals (Sweden)

    Takehito Kaneko

    Full Text Available The clustered regularly interspaced short palindromic repeat (CRISPR/CRISPR-associated (Cas system is a powerful tool for genome editing in animals. Recently, new technology has been developed to genetically modify animals without using highly skilled techniques, such as pronuclear microinjection of endonucleases. Technique for animal knockout system by electroporation (TAKE method is a simple and effective technology that produces knockout rats by introducing endonuclease mRNAs into intact embryos using electroporation. Using TAKE method and CRISPR/Cas system, the present study successfully produced knockout and knock-in mice and rats. The mice and rats derived from embryos electroporated with Cas9 mRNA, gRNA and single-stranded oligodeoxynucleotide (ssODN comprised the edited targeted gene as a knockout (67% of mice and 88% of rats or knock-in (both 33%. The TAKE method could be widely used as a powerful tool to produce genetically modified animals by genome editing.

  9. Calcium sparks in the intact gerbil spiral modiolar artery

    Directory of Open Access Journals (Sweden)

    Berge Samantha

    2011-08-01

    Full Text Available Abstract Background Calcium sparks are ryanodine receptor mediated transient calcium signals that have been shown to hyperpolarize the membrane potential by activating large conductance calcium activated potassium (BK channels in vascular smooth muscle cells. Along with voltage-dependent calcium channels, they form a signaling unit that has a vasodilatory influence on vascular diameter and regulation of myogenic tone. The existence and role of calcium sparks has hitherto been unexplored in the spiral modiolar artery, the end artery that controls blood flow to the cochlea. The goal of the present study was to determine the presence and properties of calcium sparks in the intact gerbil spiral modiolar artery. Results Calcium sparks were recorded from smooth muscle cells of intact arteries loaded with fluo-4 AM. Calcium sparks occurred with a frequency of 2.6 Hz, a rise time of 17 ms and a time to half-decay of 20 ms. Ryanodine reduced spark frequency within 3 min from 2.6 to 0.6 Hz. Caffeine (1 mM increased spark frequency from 2.3 to 3.3 Hz and prolonged rise and half-decay times from 17 to 19 ms and from 20 to 23 ms, respectively. Elevation of potassium (3.6 to 37.5 mM, presumably via depolarization, increased spark frequency from 2.4 to 3.2 Hz. Neither ryanodine nor depolarization changed rise or decay times. Conclusions This is the first characterization of calcium sparks in smooth muscle cells of the spiral modiolar artery. The results suggest that calcium sparks may regulate the diameter of the spiral modiolar artery and cochlear blood flow.

  10. Static cerebrovascular pressure autoregulation remains intact during deep hypothermia.

    Science.gov (United States)

    Goswami, Dheeraj; McLeod, Katherine; Leonard, Samantha; Kibler, Kathleen; Easley, Ronald Blaine; Fraser, Charles D; Andropoulos, Dean; Brady, Ken

    2017-09-01

    Clinical studies measuring cerebral blood flow in infants during deep hypothermia have demonstrated diminished cerebrovascular pressure autoregulation. The coexistence of hypotension in these cohorts confounds the conclusion that deep hypothermia impairs cerebrovascular pressure autoregulation. We sought to compare the lower limit of autoregulation and the static rate of autoregulation between normothermic and hypothermic piglets. Twenty anesthetized neonatal piglets (5-7 days old; 10 normothermic and 10 hypothermic to 20°C) had continuous measurements of cortical red cell flux using laser Doppler flowmetry, while hemorrhagic hypotension was induced without cardiopulmonary bypass. Lower limit of autoregulation was determined for each subject using piecewise regression and SRoR was determined above and below each lower limit of autoregulation as (%change cerebrovascular resistance/%change cerebral perfusion pressure). The estimated difference in lower limit of autoregulation was 1.4 mm Hg (lower in the hypothermic piglets; 95% C.I. -10 to 14 mm Hg; P=0.6). The median lower limit of autoregulation in the normothermic group was 39 mm Hg [IQR 38-51] vs 35 mm Hg [31-50] in the hypothermic group. Intact steady-state pressure autoregulation was defined as static rate of autoregulation >0.5 and was demonstrated in all normothermic subjects (static rate of autoregulation=0.72 [0.65-0.87]) and in 9/10 of the hypothermic subjects (static rate of autoregulation=0.65 [0.52-0.87]). This difference in static rate of autoregulation of 0.06 (95% C.I. -0.3 to 0.1) was not significant (P=0.4). Intact steady-state cerebrovascular pressure autoregulation is demonstrated in a swine model of profound hypothermia. Lower limit of autoregulation and static rate of autoregulation were similar in hypothermic and normothermic subjects. © 2017 John Wiley & Sons Ltd.

  11. Mouse models of intracranial aneurysm.

    Science.gov (United States)

    Wang, Yutang; Emeto, Theophilus I; Lee, James; Marshman, Laurence; Moran, Corey; Seto, Sai-wang; Golledge, Jonathan

    2015-05-01

    Subarachnoid hemorrhage secondary to rupture of an intracranial aneurysm is a highly lethal medical condition. Current management strategies for unruptured intracranial aneurysms involve radiological surveillance and neurosurgical or endovascular interventions. There is no pharmacological treatment available to decrease the risk of aneurysm rupture and subsequent subarachnoid hemorrhage. There is growing interest in the pathogenesis of intracranial aneurysm focused on the development of drug therapies to decrease the incidence of aneurysm rupture. The study of rodent models of intracranial aneurysms has the potential to improve our understanding of intracranial aneurysm development and progression. This review summarizes current mouse models of intact and ruptured intracranial aneurysms and discusses the relevance of these models to human intracranial aneurysms. The article also reviews the importance of these models in investigating the molecular mechanisms involved in the disease. Finally, potential pharmaceutical targets for intracranial aneurysm suggested by previous studies are discussed. Examples of potential drug targets include matrix metalloproteinases, stromal cell-derived factor-1, tumor necrosis factor-α, the renin-angiotensin system and the β-estrogen receptor. An agreed clear, precise and reproducible definition of what constitutes an aneurysm in the models would assist in their use to better understand the pathology of intracranial aneurysm and applying findings to patients.

  12. The Knockout Mouse Project

    OpenAIRE

    Austin, Christopher P.; Battey, James F.; Bradley, Allan; Bucan, Maja; Capecchi, Mario; Collins, Francis S; Dove, William F.; Duyk, Geoffrey; Dymecki, Susan; Eppig, Janan T.; Grieder, Franziska B.; Heintz, Nathaniel; Hicks, Geoff; Insel, Thomas R; Joyner, Alexandra

    2004-01-01

    Mouse knockout technology provides a powerful means of elucidating gene function in vivo, and a publicly available genome-wide collection of mouse knockouts would be significantly enabling for biomedical discovery. To date, published knockouts exist for only about 10% of mouse genes. Furthermore, many of these are limited in utility because they have not been made or phenotyped in standardized ways, and many are not freely available to researchers. It is time to harness new technologies and e...

  13. Prophylactic antibiotics for inhibiting preterm labour with intact membranes.

    Science.gov (United States)

    Flenady, Vicki; Hawley, Glenda; Stock, Owen M; Kenyon, Sara; Badawi, Nadia

    2013-12-05

    The aetiology of preterm birth is complex and there is evidence that subclinical genital tract infection influences preterm labour in some women but the role of prophylactic antibiotic treatment in the management of preterm labour is controversial. Since rupture of the membranes is an important factor in the progression of preterm labour, it is important to see if the routine administration of antibiotics confers any benefit or causes harm, prior to membrane rupture. To assess the effects of prophylactic antibiotics administered to women in preterm labour with intact membranes, on maternal and neonatal outcomes. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (31 August 2013). Randomised trials that compared antibiotic treatment with placebo or no treatment for women in preterm labour (between 20 and 36 weeks' gestation) with intact membranes. Two review authors independently assessed trial eligibility, and undertook quality assessment and data extraction. We contacted study authors for additional information. Results are presented using risk ratio (RR) for categorical data and mean difference (MD) for data measured on a continuous scale with their respective 95% confidence intervals (CI). The number needed to treat to benefit (NNTB) and the number needed to treat to harm (NNTH) was calculated where appropriate. In this update (2013), with the addition of three trials (305 women), the large ORACLE II 2001 trial continues to dominate the results of this review. This review now includes a total of 14 studies randomising 7837 women. No significant difference was shown in perinatal or infant mortality for infants of women allocated to any prophylactic antibiotics compared with no antibiotics. However, an increase in neonatal deaths was shown for infants of women receiving any prophylactic antibiotics when compared with placebo (RR 1.57, 95% CI 1.03 to 2.40; NNTH 149, 95% CI 2500 to 61). No reduction in preterm birth or other clinically

  14. Replacing the computer mouse

    OpenAIRE

    Dernoncourt, Franck

    2014-01-01

    In a few months the computer mouse will be half-a-century-old. It is known to have many drawbacks, the main ones being: loss of productivity due to constant switching between keyboard and mouse, and health issues such as RSI. Like the keyboard, it is an unnatural human-computer interface. However the vast majority of computer users still use computer mice nowadays. In this article, we explore computer mouse alternatives. Our research shows that moving the mouse cursor can be done efficiently ...

  15. An encyclopedia of mouse DNA elements (Mouse ENCODE).

    Science.gov (United States)

    Stamatoyannopoulos, John A; Snyder, Michael; Hardison, Ross; Ren, Bing; Gingeras, Thomas; Gilbert, David M; Groudine, Mark; Bender, Michael; Kaul, Rajinder; Canfield, Theresa; Giste, Erica; Johnson, Audra; Zhang, Mia; Balasundaram, Gayathri; Byron, Rachel; Roach, Vaughan; Sabo, Peter J; Sandstrom, Richard; Stehling, A Sandra; Thurman, Robert E; Weissman, Sherman M; Cayting, Philip; Hariharan, Manoj; Lian, Jin; Cheng, Yong; Landt, Stephen G; Ma, Zhihai; Wold, Barbara J; Dekker, Job; Crawford, Gregory E; Keller, Cheryl A; Wu, Weisheng; Morrissey, Christopher; Kumar, Swathi A; Mishra, Tejaswini; Jain, Deepti; Byrska-Bishop, Marta; Blankenberg, Daniel; Lajoie, Bryan R; Jain, Gaurav; Sanyal, Amartya; Chen, Kaun-Bei; Denas, Olgert; Taylor, James; Blobel, Gerd A; Weiss, Mitchell J; Pimkin, Max; Deng, Wulan; Marinov, Georgi K; Williams, Brian A; Fisher-Aylor, Katherine I; Desalvo, Gilberto; Kiralusha, Anthony; Trout, Diane; Amrhein, Henry; Mortazavi, Ali; Edsall, Lee; McCleary, David; Kuan, Samantha; Shen, Yin; Yue, Feng; Ye, Zhen; Davis, Carrie A; Zaleski, Chris; Jha, Sonali; Xue, Chenghai; Dobin, Alex; Lin, Wei; Fastuca, Meagan; Wang, Huaien; Guigo, Roderic; Djebali, Sarah; Lagarde, Julien; Ryba, Tyrone; Sasaki, Takayo; Malladi, Venkat S; Cline, Melissa S; Kirkup, Vanessa M; Learned, Katrina; Rosenbloom, Kate R; Kent, W James; Feingold, Elise A; Good, Peter J; Pazin, Michael; Lowdon, Rebecca F; Adams, Leslie B

    2012-08-13

    To complement the human Encyclopedia of DNA Elements (ENCODE) project and to enable a broad range of mouse genomics efforts, the Mouse ENCODE Consortium is applying the same experimental pipelines developed for human ENCODE to annotate the mouse genome.

  16. Capillary electrophoresis of intact basic proteins using noncovalently triple-layer coated capillaries.

    Science.gov (United States)

    Haselberg, Rob; de Jong, Gerhardus J; Somsen, Govert W

    2009-07-01

    The usefulness of a noncovalent, positively charged capillary coating for the efficient analysis of intact basic proteins with CE was studied. Capillaries were coated by subsequent flushing with solutions of 10% w/v Polybrene (PB), 3% w/v dextran sulfate (DS), and again 10% w/v PB. Coating characterization studies showed that stable coatings could be produced which exhibited a pH-independent and highly reproducible EOF. The PB-DS-PB coating was evaluated with Tris phosphate BGEs of various pH using the four basic model proteins: alpha-chymotrypsinogen A, ribonuclease A, cytochrome c, and lysozyme. Typical migration time RSDs for the proteins were less than 0.85%, and apparent plate numbers were above 125,000 using a capillary length of 40 cm. The high separation efficiency allowed detection of several minor impurities in the model proteins. Using a BGE of medium pH, the CE system with triple-layer coating appeared to be useful for the repeatable profiling of recombinant humanized mouse monoclonal immunoglobulin G(1) showing a characteristic pattern of glycoforms. The CE system was also applied to the characterization of two llama antibodies, which were produced in Saccharomyces cerevisiae, revealing the presence of a side product in one of the antibodies. The high migration time stability allowed the reliable determination of antibody-antigen binding by monitoring migration time shifts. Finally, the feasibility of using the PB-DS-PB coated capillaries for CE with mass spectrometric detection was shown by the characterization of the impure llama antibody sample.

  17. Identification of Intrinsic Axon Growth Modulators for Intact CNS Neurons after Injury

    Directory of Open Access Journals (Sweden)

    Kathren L. Fink

    2017-03-01

    Full Text Available Functional deficits persist after spinal cord injury (SCI because axons in the adult mammalian central nervous system (CNS fail to regenerate. However, modest levels of spontaneous functional recovery are typically observed after trauma and are thought to be mediated by the plasticity of intact circuitry. The mechanisms underlying intact circuit plasticity are not delineated. Here, we characterize the in vivo transcriptome of sprouting intact neurons from Ngr1 null mice after partial SCI. We identify the lysophosphatidic acid signaling modulators LPPR1 and LPAR1 as intrinsic axon growth modulators for intact corticospinal motor neurons after adjacent injury. Furthermore, in vivo LPAR1 inhibition or LPPR1 overexpression enhances sprouting of intact corticospinal tract axons and yields greater functional recovery after unilateral brainstem lesion in wild-type mice. Thus, the transcriptional profile of injury-induced sprouting of intact neurons reveals targets for therapeutic enhancement of axon growth initiation and new synapse formation.

  18. NCBI nr-aa BLAST: CBRC-MMUS-06-0131 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0131 ref|NP_444459.1| vomeronasal 1 receptor, B8 [Mus musculus] sp|Q9EQ45|VN1B8_MOUSE Vomeronas...al type-1 receptor B8 (Vomeronasal type-1 receptor A13) gb|AAG42088.1|AF291494_1 vomeronasal... receptor V1RB8 [Mus musculus] dbj|BAB79220.1| vomeronasal receptor 1 A13 [Mus musculus] gb|AAI40252.1| Vomeronasal... 1 receptor, B8 [synthetic construct] gb|EDK99274.1| vomeronasal ...1 receptor, B8 [Mus musculus] gb|AAI46525.1| Vomeronasal 1 receptor, B8 [synthetic construct] NP_444459.1 1e-175 100% ...

  19. NCBI nr-aa BLAST: CBRC-OCUN-01-0391 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OCUN-01-0391 ref|NP_444459.1| vomeronasal 1 receptor, B8 [Mus musculus] sp|Q9EQ45|VN1B8_MOUSE Vomeronas...al type-1 receptor B8 (Vomeronasal type-1 receptor A13) gb|AAG42088.1|AF291494_1 vomeronasal... receptor V1RB8 [Mus musculus] dbj|BAB79220.1| vomeronasal receptor 1 A13 [Mus musculus] gb|AAI40252.1| Vomeronasal... 1 receptor, B8 [synthetic construct] gb|EDK99274.1| vomeronasal ...1 receptor, B8 [Mus musculus] gb|AAI46525.1| Vomeronasal 1 receptor, B8 [synthetic construct] NP_444459.1 8e-37 51% ...

  20. Mechanisms of the Frank-Starling phenomena studied in intact hearts.

    Science.gov (United States)

    Burkhoff, D; Stennett, R A; Ogino, K

    1995-01-01

    The impact of ventricular volume on the relationship between intracellular calcium and ventricular pressure under steady-state conditions was determined in intact ferret hearts. The results reveal major quantitative differences and minor qualitative differences between these relations and those previously measured in isolated intact and skinned cardiac muscle. The importance of these differences is discussed within the context of developing a comprehensive mechanistic theory to describe load-dependence of the intact ventricle.

  1. Bryostatins activate protein kinase C in intact human platelets

    Energy Technology Data Exchange (ETDEWEB)

    Smith, J.B.; Tallant, E.A.; Pettit, G.R.; Wallace, R.W.

    1986-05-01

    Bryostatins, macrocyclic lactones isolated from a marine bryozoan, have antineoplastic activity in the P388 lymphocytic leukemia system. These compounds also stimulate growth in Swiss 3T3 cells, induce secretion in leukocytes, inhibit phorbol dibutyrate binding to a high affinity receptor, and activate the C-kinase in vitro. In human platelets, phorbol esters induce aggregation and activate protein kinase C, resulting in phosphorylation of a 47K protein and the 20K myosin light chain. The authors now show that bryostatin 7 (B-7) triggers platelet aggregation to the same rate and extent as phorbol 12-myristate 13-acetate (PMA). B-7 also causes the in vivo activation of the C-kinase, resulting in phosphorylation of both the 47K and the 20K proteins; the time courses and dose-responses of these B-7-induced phosphorylations were similar to those found with PMA. In addition, B-7 increases the level of /sup 32/P-incorporation into the platelet polyphosphoinositides, which also occurs in response to PMA. Bryostatin 3 (B-3), which has been shown to be much less potent than B-7 in mimicking other PMA effects, was much less effective than PMA or B-7 in inducing platelet aggregation and in stimulating /sup 32/P-incorporation into both proteins and the phosphoinositides. These results demonstrate that, intact human platelets, bryostatins mimic the phorbol esters tumor promoters and directly activate protein kinase C.

  2. Rapid isolation of intact, viable fetal cartilage models

    Energy Technology Data Exchange (ETDEWEB)

    Schmidt, R.R.; Chepenik, K.P.; Paynton, B.V.; Cotler, J.M.

    1982-04-01

    A rapid procedure is described for the isolation of viable, intact, femoral cartilage models (humeri and femora) obtained from pregnant rats on the 18th day of gestation. Viability of these models is demonstrated in an in vitro system where the incorporation of /sup 35/S-sulfate was linear with time of incubation and with numbers of cartilage models utilized. Treatment of cartilage models with ice-cold trichloroacetic acid and a boiling water bath prior to incubation with radiolabel, reduced the amount of radioactivity incorporated to 1.3% of that observed for models incubated by routine procedures. Furthermore, digestion of cartilage model homogenates with protease yielded a supernatant from which 51% to 57% of the radioactivity was precipitated as GAG. This method may also be used to isolate fetal cartilage models as early as the 16th day of gestation. with this system, specific biochemical parameters of mammalian fetal chondrogenesis may be surveyed in normally and abnormally developing fetal cartilage free of surrounding soft tissue.

  3. Compensatory mechanisms underlie intact task-switching performance in schizophrenia.

    Science.gov (United States)

    Jamadar, S; Michie, P; Karayanidis, F

    2010-04-01

    Individuals with schizophrenia tend to perform poorly on many measures of cognitive control. However, recent task-switching studies suggest that they show intact task-switching performance, despite the fact that the regions involved in task-switching are known to be structurally and functionally impaired in the disorder. Behavioral, event-related potential (ERP) and functional magnetic resonance imaging (fMRI) measures were used to compare the temporal and spatial dynamics of task-switching performance in individuals with schizophrenia and controls. Consistent with previous studies, reaction time (RT) switch cost and its reduction with anticipatory preparation did not differ between groups. There were also no group differences on cue-locked ERP components associated with anticipatory preparation processes. However, both stimulus- and response-locked ERPs were significantly disrupted in schizophrenia, suggesting difficulty with task-set implementation. fMRI analyses indicated that individuals with schizophrenia showed hyperactivity in the dorsolateral prefrontal cortex and posterior parietal cortex. RT-fMRI and ERP-fMRI associations suggested that individuals with schizophrenia employ compensatory mechanisms to overcome difficulties in task-set implementation and thereby achieve the same behavioral outcomes as controls.

  4. Spontaneous NA+ transients in individual mitochondria of intact astrocytes.

    Science.gov (United States)

    Azarias, Guillaume; Van de Ville, Dimitri; Unser, Michael; Chatton, Jean-Yves

    2008-02-01

    Mitochondria in intact cells maintain low Na(+) levels despite the large electrochemical gradient favoring cation influx into the matrix. In addition, they display individual spontaneous transient depolarizations. The authors report here that individual mitochondria in living astrocytes exhibit spontaneous increases in their Na(+) concentration (Na(mit)(+) spiking), as measured using the mitochondrial probe CoroNa Red. In a field of view with approximately 30 astrocytes, up to 1,400 transients per minute were typically detected under resting conditions. Na(mit)(+) spiking was also observed in neurons, but was scarce in two nonneural cell types tested. Astrocytic Na(mit)(+) spikes averaged 12.2 +/- 0.8 s in duration and 35.5 +/- 3.2 mM in amplitude and coincided with brief mitochondrial depolarizations; they were impaired by mitochondrial depolarization and ruthenium red pointing to the involvement of a cation uniporter. Na(mit)(+) spiking activity was significantly inhibited by mitochondrial Na(+)/H(+) exchanger inhibition and sensitive to cellular pH and Na(+) concentration. Ca(2+) played a permissive role on Na(mit)(+) spiking activity. Finally, the authors present evidence suggesting that Na(mit)(+) spiking frequency was correlated with cellular ATP levels. This study shows that, under physiological conditions, individual mitochondria in living astrocytes exhibit fast Na(+) exchange across their inner membrane, which reveals a new form of highly dynamic and localized functional regulation.

  5. Intact implicit statistical learning in borderline personality disorder.

    Science.gov (United States)

    Unoka, Zsolt; Vizin, Gabriella; Bjelik, Anna; Radics, Dóra; Nemeth, Dezso; Janacsek, Karolina

    2017-09-01

    Wide-spread neuropsychological deficits have been identified in borderline personality disorder (BPD). Previous research found impairments in decision making, declarative memory, working memory and executive functions; however, no studies have focused on implicit learning in BPD yet. The aim of our study was to investigate implicit statistical learning by comparing learning performance of 19 BPD patients and 19 healthy, age-, education- and gender-matched controls on a probabilistic sequence learning task. Moreover, we also tested whether participants retain the acquired knowledge after a delay period. To this end, participants were retested on a shorter version of the same task 24h after the learning phase. We found intact implicit statistical learning as well as retention of the acquired knowledge in this personality disorder. BPD patients seem to be able to extract and represent regularities implicitly, which is in line with the notion that implicit learning is less susceptible to illness compared to the more explicit processes. Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.

  6. Stress dependence of permeability of intact and fractured shale cores.

    Science.gov (United States)

    van Noort, Reinier; Yarushina, Viktoriya

    2016-04-01

    Whether a shale acts as a caprock, source rock, or reservoir, understanding fluid flow through shale is of major importance for understanding fluid flow in geological systems. Because of the low permeability of shale, flow is thought to be largely confined to fractures and similar features. In fracking operations, fractures are induced specifically to allow for hydrocarbon exploration. We have constructed an experimental setup to measure core permeabilities, using constant flow or a transient pulse. In this setup, we have measured the permeability of intact and fractured shale core samples, using either water or supercritical CO2 as the transporting fluid. Our measurements show decreasing permeability with increasing confining pressure, mainly due to time-dependent creep. Furthermore, our measurements show that for a simple splitting fracture, time-dependent creep will also eliminate any significant effect of this fracture on permeability. This effect of confinement on fracture permeability can have important implications regarding the effects of fracturing on shale permeability, and hence for operations depending on that.

  7. Covert spatial attention is functionally intact in amblyopic human adults.

    Science.gov (United States)

    Roberts, Mariel; Cymerman, Rachel; Smith, R Theodore; Kiorpes, Lynne; Carrasco, Marisa

    2016-12-01

    Certain abnormalities in behavioral performance and neural signaling have been attributed to a deficit of visual attention in amblyopia, a neurodevelopmental disorder characterized by a diverse array of visual deficits following abnormal binocular childhood experience. Critically, most have inferred attention's role in their task without explicitly manipulating and measuring its effects against a baseline condition. Here, we directly investigate whether human amblyopic adults benefit from covert spatial attention-the selective processing of visual information in the absence of eye movements-to the same degree as neurotypical observers. We manipulated both involuntary (Experiment 1) and voluntary (Experiment 2) attention during an orientation discrimination task for which the effects of covert spatial attention have been well established in neurotypical and special populations. In both experiments, attention significantly improved accuracy and decreased reaction times to a similar extent (a) between the eyes of the amblyopic adults and (b) between the amblyopes and their age- and gender-matched controls. Moreover, deployment of voluntary attention away from the target location significantly impaired task performance (Experiment 2). The magnitudes of the involuntary and voluntary attention benefits did not correlate with amblyopic depth or severity. Both groups of observers showed canonical performance fields (better performance along the horizontal than vertical meridian and at the lower than upper vertical meridian) and similar effects of attention across locations. Despite their characteristic low-level vision impairments, covert spatial attention remains functionally intact in human amblyopic adults.

  8. Structure of the intact ATM/Tel1 kinase.

    Science.gov (United States)

    Wang, Xuejuan; Chu, Huanyu; Lv, Mengjuan; Zhang, Zhihui; Qiu, Shuwan; Liu, Haiyan; Shen, Xuetong; Wang, Weiwu; Cai, Gang

    2016-05-27

    The ataxia-telangiectasia mutated (ATM) protein is an apical kinase that orchestrates the multifaceted DNA-damage response. Normally, ATM kinase is in an inactive, homodimer form and is transformed into monomers upon activation. Besides a conserved kinase domain at the C terminus, ATM contains three other structural modules, referred to as FAT, FATC and N-terminal helical solenoid. Here we report the first cryo-EM structure of ATM kinase, which is an intact homodimeric ATM/Tel1 from Schizosaccharomyces pombe. We show that two monomers directly contact head-to-head through the FAT and kinase domains. The tandem N-terminal helical solenoid tightly packs against the FAT and kinase domains. The structure suggests that ATM/Tel1 dimer interface and the consecutive HEAT repeats inhibit the binding of kinase substrates and regulators by steric hindrance. Our study provides a structural framework for understanding the mechanisms of ATM/Tel1 regulation as well as the development of new therapeutic agents.

  9. Measurement of the Mechanical Properties of Intact Collagen Fibrils

    Science.gov (United States)

    Mercedes, H.; Heim, A.; Matthews, W. G.; Koob, T.

    2006-03-01

    Motivated by the genetic disorder Ehlers-Danlos syndrome (EDS), in which proper collagen synthesis is interrupted, we are investigating the structural and mechanical properties of collagen fibrils. The fibrous glycoprotein collagen is the most abundant protein found in the human body and plays a key role in the extracellular matrix of the connective tissue, the properties of which are altered in EDS. We have selected as our model system the collagen fibrils of the sea cucumber dermis, a naturally mutable tissue. This system allows us to work with native fibrils which have their proteoglycan complement intact, something that is not possible with reconstituted mammalian collagen fibrils. Using atomic force microscopy, we measure, as a function of the concentration of divalent cations, the fibril diameter, its response to force loading, and the changes in its rigidity. Through these experiments, we will shed light on the mechanisms which control the properties of the sea cucumber dermis and hope to help explain the altered connective tissue extracellular matrix properties associated with EDS.

  10. Measurement of tritiated norepinephrine metabolism in intact rat brain

    Energy Technology Data Exchange (ETDEWEB)

    Levitt, M.; Kowalik, S.; Barkai, A.I. (New York State Psychiatric Inst., New York (USA))

    1983-06-01

    A procedure for the study of NE metabolism in the intact rat brain is described. The method involves ventriculocisternal perfusion of the adult male rat with artificial CSF containing (/sup 3/H)NE. Radioactivity in the perfusate associated with NE and its metabolites 3,4-dihydroxymandelic acid (DOMA), 3,4-dihydroxphenylethyleneglycol (DHPG), 3-methoxy-4-hydroxymandelic acid (VMA), 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG), and normetanephrine (NMN) is separated using high-performance liquid chromatography (HPLC). After 80 min the radioactivity in the perfusate reaches an apparent steady-state. Analysis of the steady-state samples shows higher activity in the fractions corresponding to DHPG and MHPG than in those corresponding to DOMA and VMA, confirming glycol formation as the major pathway of NE metabolism in the rat brain. Pretreatment with an MAO inhibitor (tranylcypromine) results in a marked decrease in the deaminated metabolites DHPG and MHPG and a concurrent increase in NMN. The results indicate this to be a sensitive procedure for the in vivo determination of changes in NE metabolism.

  11. HIV-2 neutralization by intact V3-specific Fab fragments

    Directory of Open Access Journals (Sweden)

    Sourial Samer

    2008-08-01

    Full Text Available Abstract The V3 region of both HIV-1 gp120 and HIV-2 gp125 surface glycoprotein has been described as a target for neutralizing antibodies. In this study a conformation-sensitive (3C4 and a linear site-specific (7C8 anti-HIV-2 V3 monoclonal antibody (mAb were characterized. The neutralization capacity of the purified mAbs and their respective papain-generated Fab fragments was analyzed. The Fabs were further characterized by sequence analysis. Our results demonstrate that neither purified mAbs were capable of neutralizing HIV-2, while intact Fab fragments from both mAbs blocked in vitro infection of HIV-2 isolates. Moreover, the conformation sensitive 3C4 Fab neutralized both subtype A and B HIV-2 isolates and SIVsm. Sequence analysis of the hypervariable regions of 3C4 Fab and 7C8 Fab revealed that the third CDR of the heavy chain (CDRH3 of the antibodies was not as long as many of the previously characterized neutralizing antibodies. Our findings suggest that whole 7C8 and 3C4 mAbs are sterically hindered from neutralizing HIV-2, whereas the smaller size of Fab fragments enables access to the V3 region on the virion surface.

  12. Visual associative learning in restrained honey bees with intact antennae.

    Directory of Open Access Journals (Sweden)

    Scott E Dobrin

    Full Text Available A restrained honey bee can be trained to extend its proboscis in response to the pairing of an odor with a sucrose reward, a form of olfactory associative learning referred to as the proboscis extension response (PER. Although the ability of flying honey bees to respond to visual cues is well-established, associative visual learning in restrained honey bees has been challenging to demonstrate. Those few groups that have documented vision-based PER have reported that removing the antennae prior to training is a prerequisite for learning. Here we report, for a simple visual learning task, the first successful performance by restrained honey bees with intact antennae. Honey bee foragers were trained on a differential visual association task by pairing the presentation of a blue light with a sucrose reward and leaving the presentation of a green light unrewarded. A negative correlation was found between age of foragers and their performance in the visual PER task. Using the adaptations to the traditional PER task outlined here, future studies can exploit pharmacological and physiological techniques to explore the neural circuit basis of visual learning in the honey bee.

  13. Management of Relocation in Cognitively Intact Older Adults.

    Science.gov (United States)

    Hertz, Judith E; Koren, Mary Elaine; Rossetti, Jeanette; Tibbits, Kathryn

    2016-11-01

    Relocation, a major life transition that can affect health positively and negatively, is moving from one permanent home to another. Many older adults will relocate at some time during their life. Relocation is also a complex process that requires careful consideration and planning before the move (i.e., pre-location) and adjustment to the new home after the move (i.e., post-relocation). The current article is a summary of content based on a comprehensive evidence-based practice guideline focused on management of relocation in cognitively intact older adults. The guideline was designed to be used across diverse settings by nurses and other providers. Pre-relocation guidelines include assessment for the need for relocation, interventions prior to moving, and outcomes for evaluation of the pre-relocation process. For post-relocation, content focuses on assessment of risks for not adjusting after the move as well as intervention guidelines to promote adjustment and outcomes for evaluation. Implications include advocacy for older adults by using the guideline, disseminating it, and conducting future research. [Journal of Gerontological Nursing, 42(11), 14-23.].

  14. Activation and deactivation of vibronic channels in intact phycocyanin rods

    Science.gov (United States)

    Nganou, C.; David, L.; Meinke, R.; Adir, N.; Maultzsch, J.; Mkandawire, M.; Pouhè, D.; Thomsen, C.

    2014-02-01

    We investigated the excitation modes of the light-harvesting protein phycocyanin (PC) from Thermosynechococcus vulcanus in the crystalline state using UV and near-infrared Raman spectroscopy. The spectra revealed the absence of a hydrogen out-of-plane wagging (HOOP) mode in the PC trimer, which suggests that the HOOP mode is activated in the intact PC rod, while it is not active in the PC trimer. Furthermore, in the PC trimer an intense mode at 984 cm-1 is assigned to the C-C stretching vibration while the mode at 454 cm-1 is likely due to ethyl group torsion. In contrast, in the similar chromophore phytochromobilin the C5,10,15-D wag mode at 622 cm-1 does not come from a downshift of the HOOP. Additionally, the absence of modes between 1200 and 1300 cm-1 rules out functional monomerization. A correlation between phycocyanobilin (PCB) and phycoerythrobilin (PEB) suggests that the PCB cofactors of the PC trimer appear in a conformation similar to that of PEB. The conformation of the PC rod is consistent with that of the allophycocyanin (APC) trimer, and thus excitonic flow is facilitated between these two independent light-harvesting compounds. This excitonic flow from the PC rod to APC appears to be modulated by the vibration channels during HOOP wagging, C = C stretching, and the N-H rocking in-plan vibration.

  15. Amniotic Fluid Infection in Preterm Pregnancies with Intact Membranes

    Science.gov (United States)

    Rahkonen, Leena; Nupponen, Irmeli; Pätäri-Sampo, Anu; Tikkanen, Minna; Sorsa, Timo; Juhila, Juuso; Andersson, Sture; Paavonen, Jorma; Stefanovic, Vedran

    2017-01-01

    Introduction. Intra-amniotic infection (IAI) is a major cause of preterm labor and adverse neonatal outcome. We evaluated amniotic fluid (AF) proteolytic cascade forming biomarkers in relation to microbial invasion of the amniotic cavity (MIAC) and IAI in preterm pregnancies with intact membranes. Material and Methods. Amniocentesis was made to 73 women with singleton pregnancies; 27 with suspected IAI; and 46 controls. AF biomarkers were divided into three cascades: Cascade 1: matrix metalloproteinase-8 (MMP-8), MMP-9, myeloperoxidase (MPO), and interleukin-6; Cascade 2: neutrophil elastase (HNE), elafin, and MMP-9; Cascade 3: MMP-2, tissue inhibitor of matrix metalloproteinases-1 (TIMP-1), MMP-8/TIMP-1 molar ratio, and C-reactive protein (CRP). MMP-8 was measured by an immunoenzymometric assay and the others were measured by ELISA. Standard biochemical methods, molecular microbiology, and culture techniques were used. Results. MMP-8, MMP-9, MPO, elafin, and TIMP-1 concentrations were higher in IAI suspected cases compared to controls and also in IAI suspected cases with MIAC compared to those without MIAC when adjusted by gestational age at amniocentesis. All biomarkers except elafin and MMP-2 had the sensitivity of 100% with thresholds based on ROC-curve. Odd ratios of biomarkers for MIAC were 1.2-38 and 95% confidential intervals 1.0-353.6. Conclusions. Neutrophil based AF biomarkers were associated with IAI and MIAC. PMID:28167848

  16. The MOUSE Squad

    Science.gov (United States)

    Borja, Rhea R.

    2004-01-01

    This article presents a New York city after-school program started by MOUSE (Making Opportunities for Upgrading Schools and Education), a national nonprofit group that teaches students how to fix computers, and equips them with the communication and problem-solving skills to help them in the working world. The MOUSE program is part of a trend…

  17. DNA sequences within glioma-derived extracellular vesicles can cross the intact blood-brain barrier and be detected in peripheral blood of patients

    Science.gov (United States)

    Lázaro-Ibáñez, Elisa; Peris-Celda, María; Alonso, Marta M.; Guzmán-De-Villoria, Juan; Fernández-Carballal, Carlos; de Mendivil, Ana Ortiz; García-Duque, Sara; Escobedo-Lucea, Carmen; Prat-Acín, Ricardo; Belda-Iniesta, Cristóbal; Ayuso-Sacido, Angel

    2017-01-01

    Tumor-cell-secreted extracellular vesicles (EVs) can cross the disrupted blood-brain barrier (BBB) into the bloodstream. However, in certain gliomas, the BBB remains intact, which might limit EVs release. To evaluate the ability of tumor-derived EVs to cross the BBB, we used an orthotopic xenotransplant mouse model of human glioma-cancer stem cells featuring an intact BBB. We demonstrated that all types of tumor cells-derived EVs−apoptotic bodies, shedding microvesicles and exosomes−cross the intact BBB and can be detected in the peripheral blood, which provides a minimally invasive method for their detection compared to liquid biopsies obtained from cerebrospinal fluid (CSF). Furthermore, these EVs can be readily distinguished from total murine EVs, since they carry human-specific DNA sequences relevant for GBM biology. In a small cohort of glioma patients, we finally demonstrated that peripheral blood EVs cargo can be successfully used to detect the presence of IDH1G395A, an essential biomarker in the current management of human glioma PMID:27902458

  18. Mouse genome database 2016.

    Science.gov (United States)

    Bult, Carol J; Eppig, Janan T; Blake, Judith A; Kadin, James A; Richardson, Joel E

    2016-01-01

    The Mouse Genome Database (MGD; http://www.informatics.jax.org) is the primary community model organism database for the laboratory mouse and serves as the source for key biological reference data related to mouse genes, gene functions, phenotypes and disease models with a strong emphasis on the relationship of these data to human biology and disease. As the cost of genome-scale sequencing continues to decrease and new technologies for genome editing become widely adopted, the laboratory mouse is more important than ever as a model system for understanding the biological significance of human genetic variation and for advancing the basic research needed to support the emergence of genome-guided precision medicine. Recent enhancements to MGD include new graphical summaries of biological annotations for mouse genes, support for mobile access to the database, tools to support the annotation and analysis of sets of genes, and expanded support for comparative biology through the expansion of homology data.

  19. Isolation, culture and adenoviral transduction of parietal cells from mouse gastric mucosa

    Energy Technology Data Exchange (ETDEWEB)

    Gliddon, Briony L; Nguyen, Nhung V; Gunn, Priscilla A; Gleeson, Paul A; Driel, Ian R van [The Department of Biochemistry and Molecular Biology and Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, 30 Flemington Road, Parkville, Melbourne, Victoria 3010 (Australia)], E-mail: i.vandriel@unimelb.edu.au

    2008-09-01

    Here we describe a method for the isolation of intact gastric glands from mice and primary culture and transfection of mouse gastric epithelial cells. Collagenase digestion of PBS-perfused mouse stomachs released large intact gastric glands that were plated on a basement membrane matrix. The heterogeneous gland cell cultures typically contain {approx}60% parietal cells. Isolated mouse parietal cells remain viable in culture for up to 5 days and react strongly with an antibody specific to the gastric H{sup +}/K{sup +} ATPase. Isolated intact mouse gastric glands and primary cultures of mouse parietal cells respond to the secretagogue, histamine. Typical morphological changes from a resting to an acid-secreting active parietal cell were observed. In resting cultures of mouse parietal cells, the H{sup +}/K{sup +} ATPase displayed a cytoplasmic punctate staining pattern consistent with tubulovesicle element structures. Following histamine stimulation, an expansion of internal apical vacuole structures was observed together with a pronounced redistribution of the H{sup +}/K{sup +} ATPase from the cytoplasm to the apical vacuoles. A reproducible procedure to express genes of interest exogenously in these cultures of mouse parietal cells was also established. This method combines recombinant adenoviral transduction with magnetic field-assisted transfection resulting in {approx}30% transduced parietal cells. Adenoviral-transduced parietal cells maintain their ability to undergo agonist-induced activation. This protocol will be useful for the isolation, culture and expression of genes in parietal cells from genetically modified mice and as such will be an invaluable tool for studying the complex exocytic and endocytic trafficking events of the H{sup +}/K{sup +} ATPase which underpin the regulation of acid secretion.

  20. NIH Mouse Metabolic Phenotyping Centers: the power of centralized phenotyping

    Science.gov (United States)

    Kent Lloyd, K. C.; Cline, Gary W.; Wasserman, David H.

    2013-01-01

    The Mouse Metabolic Phenotyping Centers (MMPCs) were founded in 2001 by the National Institutes of Health (NIH) to advance biomedical research by providing the scientific community with standardized, high-quality phenotyping services for mouse models of diabetes, obesity, and their complications. The intent is to allow researchers to take optimum advantage of the many new mouse models produced in labs and in high-throughput public efforts. The six MMPCs are located at universities around the country and perform complex metabolic tests in intact mice and hormone and analyte assays in tissues on a fee-for-service basis. Testing is subsidized by the NIH in order to reduce the barriers for mouse researchers. Although data derived from these tests belong to the researcher submitting mice or tissues, these data are archived after publication in a public database run by the MMPC Coordinating and Bioinformatics Unit. It is hoped that data from experiments performed in many mouse models of metabolic diseases, using standard protocols, will be useful in understanding the nature of these complex disorders. The current areas of expertise include energy balance and body composition, insulin action and secretion, whole-body and tissue carbohydrate and lipid metabolism, cardiovascular and renal function, and metabolic pathway kinetics. In addition to providing services, the MMPC staff provides expertise and advice to researchers, and works to develop and refine test protocols to best meet the community’s needs in light of current scientific developments. Test technology is disseminated by publications and through annual courses. PMID:22940748

  1. Boron and Zinc Transport Through Intact Columns of Calcareous Soils

    Institute of Scientific and Technical Information of China (English)

    M. MAHMOOD-UL-HASSAN; M. S. AKHTAR; G. NABI

    2008-01-01

    Leaching of boron (B) and zinc (Zn) can be significant in some pedomorphic conditions, which can cause contamination of shallow groundwater and economic losses. Boron and Zn adsorption and transport was studied using 8.4 cm diameter ×28 cm long intact columns from two calcareous soil series with differing clay contents and vadose zone structures:Lyallpur soil series, clay loam (fine-silty, mixed, hyperthermic Ustalfic Haplargid), and Sultanpur soil series, sandy loam (coarse-silty, mixed, hyperthermic Ustollic Camborthid). The adsorption isotherms were developed by equilibrating soil with 0.01 mol L-1 CaCl2 aqueous solution containing varying amounts of B and Zn and were fitted to the Langmuir equation. The B and Zn breakthrough curves were fitted to the two-domain convective-dispersive equation. At the end of the leaching experiment, 0.11 L 10 g L-1 blue dye solution was also applied to each column to mark the flow paths.The Lyallpur soil columns had a slightly greater adsorption partition coefficient both for B and Zn than the Sultanpur soil columns. In the Lyallpur soil columns, B arrival was immediate but the peak concentration ratio (the concentration in solution at equilibrium/concentration applied) was lower than that in the Sultanpur soil columns. The breakthrough of B in the Sultanpur soil columns occurred after about 10 cm of cumulative drainage in both the columns; the rise in effluent concentration was fast and the peak concentration ratio was almost 1. Zinc leaching through the soil columns was very limited as only one column from the Lyallpur soil series showed Zn breakthrough in the effluent where the peak concentration ratio was only 0.05. This study demonstrates the effect of soil structure on B transport and has implications for the nutrient management in field soils.

  2. Raman mapping of intact biofilms on stainless steel surfaces

    Science.gov (United States)

    Nguyen, Julie K.; Heighton, Lynne; Xu, Yunfeng; Nou, Xiangwu; Schmidt, Walter F.

    2016-05-01

    Many issues occur when microbial bacteria contaminates human food or water; it can be dangerous to the public. Determining how the microbial are growing, it can help experts determine how to prevent the outbreaks. Biofilms are a tightly group of microbial cells that grow on living surfaces or surrounding themselves. Though biofilms are not necessarily uniform; when there are more than one type of microbial bacteria that are grown, Raman mapping is performed to determine the growth patterns. Depending on the type of microbial bacteria, they can grow in various patterns such as symmetrical or scattered on the surface. The biofilms need to be intact in order to preclude and potentially figuring out the relative intensity of different components in a biofilm mixture. In addition, it is important to determine whether one biofilms is a substrate for another biofilm to be detected. For example, it is possible if layer B appears above layer A, but layer A doesn't appear above layer B. In this case, three types of biofilms that are grown includes Listeria(L), Ralstonia(R), and a mixture of the two (LR). Since microbe deposits on metal surfaces are quite suitable, biofilms were grown on stainless steel surface slides. Each slide was viewed under a Raman Microscope at 100X and using a 532nm laser to provide great results and sharp peaks. The mapping of the laser helps determine how the bacteria growth, at which intensity the bacteria appeared in order to identify specific microbes to signature markers on biofilms.

  3. The effects of silver nanoparticles on intact wastewater biofilms

    Directory of Open Access Journals (Sweden)

    Zhiya eSheng

    2015-07-01

    Full Text Available Silver nanoparticles (Ag-NPs have strong antibacterial properties, which may adversely affect biological wastewater treatment processes. To determine the overall effect, intact biofilm samples were collected from the rotating biological contactor (RBC at the local wastewater treatment plant and treated with 200 mg Ag/L Ag-NPs for 24 h. The biofilm uptake of Ag-NPs was monitored with transmission electron microscopy (TEM. Forty-five min after Ag-NP application, Ag-NPs were seen in the biofilm extracellular polymeric substances (EPS. After 24 h, Ag-NPs had entered certain microbial cells, while other cells contained no observable Ag-NPs. Some cells were dying after the uptake of Ag-NPs. However, there was no significant reduction in cultivable bacteria in the biofilms, based on heterotrophic plate counts (HPC. While this may indicate that wastewater biofilms are highly resistant to Ag-NPs, the HPC represents only a small portion of the total microbial population. To further investigate the effects of Ag-NPs, a GeoChip microarray was used to directly detect changes in the functional gene structure of the microbial community in the biofilm. A clear decrease (34.6% decrease in gene number in gene diversity was evident in the GeoChip analysis. However, the complete loss of any specific gene was rare. Some gene families present in both treated and untreated biofilms. However, this doesn’t necessarily mean that there was no change in these families. Signal intensity decreased in certain variants in each family while other variants increased to compensate the effects of Ag-NPs. The results indicate that Ag-NP treatment decreased microbial community diversity but did not significantly affect the microbial community function. This provides direct evidence for the functional redundancy of microbial community in engineered ecosystems such as wastewater biofilms.

  4. Measurements and interpretations of light scattering from intact biological cells

    Science.gov (United States)

    Wilson, Jeremy D.

    Visible light interacts with biological cells primarily through elastic scattering. The details of how cells scatter light depend on their morphology and their substructures. In this thesis we first present a series of experiments and models to discern the specific contributions of certain sub-cellular constituents to whole-cell scattering. Exploiting the findings of those studies, we report on experiments within model systems of cell death that demonstrate the potential of light scattering measurements as a tool in modern biology. Instrumentation capable of exploiting the findings of this thesis from a biology-relevant microscopy platform is designed and developed. A Mie theory based interpretation of light scattering signals originating from a collection of particles with a broad size distribution is developed. Upon applying this model to scattering data from intact cells, we find that it robustly extracts the size scale of dominant light scattering particles, suggests that scattering measurements are sensitive primarily to mitochondrial and lysosomal morphology, and unites conflicting results in the literature. Using this model as a basis, we present a collection of studies in which we use various strategies of photodynamic therapy (PDT) as a biophysical tool to perturb mitochondria and lysosomes, and observe the effects of these perturbations on whole-cell scattering. Through these experiments, we are able to discern the individual contributions of mitochondria and lysosomes to whole-cell light scattering, and demonstrate that mitochondria are responsible for roughly 80% of the scattering signal. Results of experiments aimed at demonstrating the potential role that light scattering measurements have to play in future studies of cell death biology are presented. We first show that mitochondrial-PDT-induced morphology changes measured with light scattering map into the cell killing efficacy of the therapy. We next demonstrate that mitochondrial

  5. Mouse Genome Informatics (MGI)

    Data.gov (United States)

    U.S. Department of Health & Human Services — MGI is the international database resource for the laboratory mouse, providing integrated genetic, genomic, and biological data to facilitate the study of human...

  6. Mouse Phenome Database (MPD)

    Data.gov (United States)

    U.S. Department of Health & Human Services — The Mouse Phenome Database (MPD) has characterizations of hundreds of strains of laboratory mice to facilitate translational discoveries and to assist in selection...

  7. Visual speech fills in both discrimination and identification of non-intact auditory speech in children.

    Science.gov (United States)

    Jerger, Susan; Damian, Markus F; McAlpine, Rachel P; Abdi, Hervé

    2017-07-20

    To communicate, children must discriminate and identify speech sounds. Because visual speech plays an important role in this process, we explored how visual speech influences phoneme discrimination and identification by children. Critical items had intact visual speech (e.g. bæz) coupled to non-intact (excised onsets) auditory speech (signified by /-b/æz). Children discriminated syllable pairs that differed in intactness (i.e. bæz:/-b/æz) and identified non-intact nonwords (/-b/æz). We predicted that visual speech would cause children to perceive the non-intact onsets as intact, resulting in more same responses for discrimination and more intact (i.e. bæz) responses for identification in the audiovisual than auditory mode. Visual speech for the easy-to-speechread /b/ but not for the difficult-to-speechread /g/ boosted discrimination and identification (about 35-45%) in children from four to fourteen years. The influence of visual speech on discrimination was uniquely associated with the influence of visual speech on identification and receptive vocabulary skills.

  8. Brine Inclusions Migration in Intact Salt Crystals under Thermal Gradient

    Science.gov (United States)

    Caporuscio, F.; Boukhalfa, H.

    2013-12-01

    The behavior of water contained in rock salt under the influence of thermal gradients is critical to the performance of salt as a medium for the disposal of nuclear waste. Water contained in salt can be present as discrete inclusions within intact salt crystals, at the interface between salt crystals and aggregates, and also as hydration water and structural water present in accessory minerals present in salt. Water content in pure halite salt usually rages from 0.1 to 0.5 wt. % but is significantly higher in clay rich salt, for which water content can be up to several wt. %. Under the influence of thermal gradients brine inclusions and water associated to the accessory mineral is mobilized. Previous investigations have shown brine inclusions tend to move towards the heat source through a mechanism that involves the dissolution of salt at the hot face of the brine inclusion and its precipitation at the colder side of the inclusion. Uncertainties remain on the exact parameters that define the rate of brine migration and whether it truly migrates to towards the heat source. We performed studies under controlled thermal gradients to examine the behavior of brine inclusions in single salt crystals obtained from the underground salt mine at the Waste Isolation Power Plant (WIPP). We found that the behavior of the brine inclusions under thermal gradients is dependent on the thermal gradient magnitude and the nature of the inclusion. Full inclusions (liquid only) migrate predominantly towards the heat source, but when the inclusions are large and close to the surface they fracture the salt and release water near the surface. Inclusions that migrate towards the heat source migrate through a mechanism that involves the dissolution of salt at the hot side of the inclusion and its deposition along the migration path. SEM analysis of the migration pathways shows that brine migrates through the creation of a network of square shaped hollow channels of about 10 micron diameter

  9. Proteolysis breaks tolerance toward intact α345(IV) collagen, eliciting novel anti-glomerular basement membrane autoantibodies specific for α345NC1 hexamers.

    Science.gov (United States)

    Olaru, Florina; Wang, Xu-Ping; Luo, Wentian; Ge, Linna; Miner, Jeffrey H; Kleinau, Sandra; Geiger, Xochiquetzal J; Wasiluk, Andrew; Heidet, Laurence; Kitching, A Richard; Borza, Dorin-Bogdan

    2013-02-15

    Goodpasture disease is an autoimmune kidney disease mediated by autoantibodies against noncollagenous domain 1 (NC1) monomers of α3(IV) collagen that bind to the glomerular basement membrane (GBM), usually causing rapidly progressive glomerulonephritis (GN). We identified a novel type of human IgG4-restricted anti-GBM autoantibodies associated with mild nonprogressive GN, which specifically targeted α345NC1 hexamers but not α3NC1 monomers. The mechanisms eliciting these anti-GBM autoantibodies were investigated in mouse models recapitulating this phenotype. Wild-type and FcγRIIB(-/-) mice immunized with autologous murine GBM NC1 hexamers produced mouse IgG1-restricted autoantibodies specific for α345NC1 hexamers, which bound to the GBM in vivo but did not cause GN. In these mice, intact collagen IV from murine GBM was not immunogenic. However, in Col4a3(-/-) Alport mice, both intact collagen IV and NC1 hexamers from murine GBM elicited IgG Abs specific for α345NC1 hexamers, which were not subclass restricted. As heterologous Ag in COL4A3-humanized mice, murine GBM NC1 hexamers elicited mouse IgG1, IgG2a, and IgG2b autoantibodies specific for α345NC1 hexamers and induced anti-GBM Ab GN. These findings indicate that tolerance toward autologous intact α345(IV) collagen is established in hosts expressing this Ag, even though autoreactive B cells specific for α345NC1 hexamers are not purged from their repertoire. Proteolysis selectively breaches this tolerance by generating autoimmunogenic α345NC1 hexamers. This provides a mechanism eliciting autoantibodies specific for α345NC1 hexamers, which are restricted to noninflammatory IgG subclasses and are nonnephritogenic. In Alport syndrome, lack of tolerance toward α345(IV) collagen promotes production of alloantibodies to α345NC1 hexamers, including proinflammatory IgG subclasses that mediate posttransplant anti-GBM nephritis.

  10. Proteolysis breaks tolerance toward intact α345(IV) collagen, eliciting novel anti-GBM autoantibodies specific for α345NC1 hexamers

    Science.gov (United States)

    Olaru, Florina; Wang, Xu-Ping; Luo, Wentian; Ge, Linna; Miner, Jeffrey H; Kleinau, Sandra; Geiger, Xochiquetzal J.; Wasiluk, Andrew; Heidet, Laurence; Kitching, A. Richard; Borza, Dorin-Bogdan

    2012-01-01

    Goodpasture disease is an autoimmune kidney disease mediated by autoAbs against NC1 monomers of α3(IV) collagen that bind to the glomerular basement membrane (GBM), usually causing rapidly progressive glomerulonephritis. We identified a novel type of human IgG4-restricted anti-GBM autoAbs associated with mild non-progressive glomerulonephritis, which specifically targeted α345NC1 hexamers but not α3NC1 monomers. The mechanisms eliciting these anti-GBM autoAbs were investigated in mouse models recapitulating this phenotype. Wild type and FcγRIIB−/− mice immunized with autologous murine GBM NC1 hexamers produced mouse IgG1-restricted autoAbs specific for α345NC1 hexamers, which bound to the GBM in vivo but did not cause glomerulonephritis. In these mice, intact collagen IV from murine GBM was not immunogenic. However, in Col4a3−/− Alport mice, both intact collagen IV and NC1 hexamers from murine GBM elicited IgG antibodies specific for α3α4α5NC1 hexamers, which were not subclass restricted. As heterologous antigen in COL4A3-humanized mice, murine GBM NC1 hexamers elicited mouse IgG1, IgG2a and IgG2b autoAbs specific for α345NC1 hexamers and induced anti-GBM Ab glomerulonephritis. These findings indicate that tolerance toward autologous intact α3α4α5(IV) collagen is established in hosts expressing this antigen, even though autoreactive B cells specific for α345NC1 hexamers are not purged from their repertoire. Proteolysis selectively breaches this tolerance by generating autoimmunogenic α3α4α5NC1 hexamers. This provides a mechanism eliciting autoAbs specific for α345NC1 hexamers, which are restricted to non-inflammatory IgG subclasses and non-nephritogenic. In Alport syndrome, lack of tolerance toward α3α4α5(IV) collagen promotes production of alloantibodies to α345NC1 hexamers, including pro-inflammatory IgG subclasses which mediate post-transplant anti-GBM nephritis. PMID:23303673

  11. Autoradiographic visualization of the mouse egg's sperm receptor bound to sperm

    OpenAIRE

    1986-01-01

    The extracellular coat, or zona pellucida, of mammalian eggs contains species-specific receptors to which sperm bind as a prelude to fertilization. In mice, ZP3, one of only three zona pellucida glycoproteins, serves as sperm receptor. Acrosome-intact, but not acrosome-reacted, mouse sperm recognize and interact with specific O- linked oligosaccharides of ZP3 resulting in sperm-egg binding. Binding, in turn, causes sperm to undergo the acrosome reaction; a membrane fusion event that results i...

  12. Colorado Plateau Rapid Ecoregion Assessment Terrestrial Intactness and Potential For Change

    Data.gov (United States)

    Bureau of Land Management, Department of the Interior — This map shows current and near-term terrestrial intactness, as well as long term potential for development and climate change. These datasets are the results of a...

  13. Colorado Plateau Rapid Ecoregion Assessment Terrestrial Intactness and Potential For Change (HUC5)

    Data.gov (United States)

    Bureau of Land Management, Department of the Interior — This map shows current and near-term terrestrial intactness, as well as long term potential for development and climate change. These datasets are the results of a...

  14. Spectral sensitivity of light induced respiratory activity of photoreceptor mitochondria in the intact fly

    NARCIS (Netherlands)

    Tinbergen, J.; Stavenga, D.G.

    1987-01-01

    Fly Calliphora erythrocephala (white eyed) photoreceptors were investigated in intact, living animals by microspectrofluorometry in vivo. The fluorescence of mitochondrial flavoproteins was used to monitor transient changes in oxidative metabolism, which were induced by a test light following a stim

  15. Ultra fast magic angle spinning solid - state NMR spectroscopy of intact bone.

    Science.gov (United States)

    Singh, Chandan; Rai, Ratan Kumar; Kayastha, Arvind M; Sinha, Neeraj

    2016-02-01

    Ultra fast magic angle spinning (MAS) has been a potent method to significantly average out homogeneous/inhomogeneous line broadening in solid-state nuclear magnetic resonance (ssNMR) spectroscopy. It has given a new direction to ssNMR spectroscopy with its different applications. We present here the first and foremost application of ultra fast MAS (~60 kHz) for ssNMR spectroscopy of intact bone. This methodology helps to comprehend and elucidate the organic content in the intact bone matrix with resolution and sensitivity enhancement. At this MAS speed, amino protons from organic part of intact bone start to appear in (1) H NMR spectra. The experimental protocol of ultra-high speed MAS for intact bone has been entailed with an additional insight achieved at 60 kHz.

  16. Colorado Plateau Rapid Ecoregion Assessment Conservation Elements: Aquatic Intactness (HUC5)

    Data.gov (United States)

    Bureau of Land Management, Department of the Interior — This map provides an estimate of current and near-term aquatic intactness, which is based on the results of a fuzzy logic model integrating land use, water quality,...

  17. High resolution TOF MS coupled to CE for the analysis of isotopically resolved intact proteins.

    Science.gov (United States)

    Taichrib, Angelina; Pelzing, Matthias; Pellegrino, Cristoforo; Rossi, Mara; Neusüss, Christian

    2011-06-10

    Intact protein analysis by mass spectrometry is of great interest for the characterisation of biotechnological products. Exact mass measurement in combination with isotopic resolution allows the detection of modifications leading to small mass changes like deamidation or reduction of disulfide bonds directly on the level of the intact protein. Here, a concept is presented based on time-of-flight mass spectrometry. A bench top TOF MS and a high resolution TOF MS were used to resolve the isotopes of intact recombinant human growth hormone and intact human erythropoietin, respectively. Thus, these 22 and around 30kDa large proteins can be characterised sensitively in great detail and along with capillary electrophoretic separation unambiguous identification of minor protein modifications like deamidation is possible. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. Mouse bladder wall injection.

    Science.gov (United States)

    Fu, Chi-Ling; Apelo, Charity A; Torres, Baldemar; Thai, Kim H; Hsieh, Michael H

    2011-07-12

    Mouse bladder wall injection is a useful technique to orthotopically study bladder phenomena, including stem cell, smooth muscle, and cancer biology. Before starting injections, the surgical area must be cleaned with soap and water and antiseptic solution. Surgical equipment must be sterilized before use and between each animal. Each mouse is placed under inhaled isoflurane anesthesia (2-5% for induction, 1-3% for maintenance) and its bladder exposed by making a midline abdominal incision with scissors. If the bladder is full, it is partially decompressed by gentle squeezing between two fingers. The cell suspension of interest is intramurally injected into the wall of the bladder dome using a 29 or 30 gauge needle and 1 cc or smaller syringe. The wound is then closed using wound clips and the mouse allowed to recover on a warming pad. Bladder wall injection is a delicate microsurgical technique that can be mastered with practice.

  19. Responses of intact and injured sural nerve fibers to cooling and menthol.

    Science.gov (United States)

    Teliban, Alina; Bartsch, Fabian; Struck, Marek; Baron, Ralf; Jänig, Wilfrid

    2014-05-01

    Intact and injured cutaneous C-fibers in the rat sural nerve are cold sensitive, heat sensitive, and/or mechanosensitive. Cold-sensitive fibers are either low-threshold type 1 cold sensitive or high-threshold type 2 cold sensitive. The hypothesis was tested, in intact and injured afferent nerve fibers, that low-threshold cold-sensitive afferent nerve fibers are activated by the transient receptor potential melastatin 8 (TRPM8) agonist menthol, whereas high-threshold cold-sensitive C-fibers and cold-insensitive afferent nerve fibers are menthol insensitive. In anesthetized rats, activity was recorded from afferent nerve fibers in strands isolated from the sural nerve, which was either intact or crushed 6-12 days before the experiment distal to the recording site. In all, 77 functionally identified afferent C-fibers (30 intact fibers, 47 injured fibers) and 34 functionally characterized A-fibers (11 intact fibers, 23 injured fibers) were tested for their responses to menthol applied to their receptive fields either in the skin (10 or 20%) or in the nerve (4 or 8 mM). Menthol activated all intact (n = 12) and 90% of injured (n = 20/22) type 1 cold-sensitive C-fibers; it activated no intact type 2 cold-sensitive C-fibers (n = 7) and 1/11 injured type 2 cold-sensitive C-fibers. Neither intact nor injured heat- and/or mechanosensitive cold-insensitive C-fibers (n = 25) and almost no A-fibers (n = 2/34) were activated by menthol. These results strongly argue that cutaneous type 1 cold-sensitive afferent fibers are nonnociceptive cold fibers that use the TRPM8 transduction channel.

  20. Structural Studies on Intact Clostridium Botulinum Neurotoxins Complexed with Inhibitors Leading to Drug Design

    Science.gov (United States)

    2006-02-01

    structure1. Introduction Tetanus neurotoxin (TeNT) produced by Clostridium tetani and the seven antigenically distinct botulinum neurotoxins (BoNT/A-G...2-0011 TITLE: Structural Studies on Intact Clostridium Botulinum Neurotoxins Complexed with Inhibitors Leading to Drug...DATES COVERED (From - To) 28 Jan 2005 – 27 Jan 2006 4. TITLE AND SUBTITLE Structural Studies on Intact Clostridium Botulinum Neurotoxins Complexed

  1. 40 CFR 261.41 - Notification and Recordkeeping for Used, Intact Cathode Ray Tubes (CRTs) Exported for Reuse.

    Science.gov (United States)

    2010-07-01

    ... Used, Intact Cathode Ray Tubes (CRTs) Exported for Reuse. 261.41 Section 261.41 Protection of... Tubes (CRTs) Exported for Reuse. (a) Persons who export used, intact CRTs for reuse must send a one-time... to export used, intact CRTs for reuse, the notifier's name, address, and EPA ID number (if...

  2. Investigation of mitochondrial dysfunction by sequential microplate-based respiration measurements from intact and permeabilized neurons.

    Directory of Open Access Journals (Sweden)

    Pascaline Clerc

    Full Text Available Mitochondrial dysfunction is a component of many neurodegenerative conditions. Measurement of oxygen consumption from intact neurons enables evaluation of mitochondrial bioenergetics under conditions that are more physiologically realistic compared to isolated mitochondria. However, mechanistic analysis of mitochondrial function in cells is complicated by changing energy demands and lack of substrate control. Here we describe a technique for sequentially measuring respiration from intact and saponin-permeabilized cortical neurons on single microplates. This technique allows control of substrates to individual electron transport chain complexes following permeabilization, as well as side-by-side comparisons to intact cells. To illustrate the utility of the technique, we demonstrate that inhibition of respiration by the drug KB-R7943 in intact neurons is relieved by delivery of the complex II substrate succinate, but not by complex I substrates, via acute saponin permeabilization. In contrast, methyl succinate, a putative cell permeable complex II substrate, failed to rescue respiration in intact neurons and was a poor complex II substrate in permeabilized cells. Sequential measurements of intact and permeabilized cell respiration should be particularly useful for evaluating indirect mitochondrial toxicity due to drugs or cellular signaling events which cannot be readily studied using isolated mitochondria.

  3. Investigation of mitochondrial dysfunction by sequential microplate-based respiration measurements from intact and permeabilized neurons.

    Science.gov (United States)

    Clerc, Pascaline; Polster, Brian M

    2012-01-01

    Mitochondrial dysfunction is a component of many neurodegenerative conditions. Measurement of oxygen consumption from intact neurons enables evaluation of mitochondrial bioenergetics under conditions that are more physiologically realistic compared to isolated mitochondria. However, mechanistic analysis of mitochondrial function in cells is complicated by changing energy demands and lack of substrate control. Here we describe a technique for sequentially measuring respiration from intact and saponin-permeabilized cortical neurons on single microplates. This technique allows control of substrates to individual electron transport chain complexes following permeabilization, as well as side-by-side comparisons to intact cells. To illustrate the utility of the technique, we demonstrate that inhibition of respiration by the drug KB-R7943 in intact neurons is relieved by delivery of the complex II substrate succinate, but not by complex I substrates, via acute saponin permeabilization. In contrast, methyl succinate, a putative cell permeable complex II substrate, failed to rescue respiration in intact neurons and was a poor complex II substrate in permeabilized cells. Sequential measurements of intact and permeabilized cell respiration should be particularly useful for evaluating indirect mitochondrial toxicity due to drugs or cellular signaling events which cannot be readily studied using isolated mitochondria.

  4. Identification of Intrinsic Axon Growth Modulators for Intact CNS Neurons after Injury.

    Science.gov (United States)

    Fink, Kathren L; López-Giráldez, Francesc; Kim, In-Jung; Strittmatter, Stephen M; Cafferty, William B J

    2017-03-14

    Functional deficits persist after spinal cord injury (SCI) because axons in the adult mammalian central nervous system (CNS) fail to regenerate. However, modest levels of spontaneous functional recovery are typically observed after trauma and are thought to be mediated by the plasticity of intact circuitry. The mechanisms underlying intact circuit plasticity are not delineated. Here, we characterize the in vivo transcriptome of sprouting intact neurons from Ngr1 null mice after partial SCI. We identify the lysophosphatidic acid signaling modulators LPPR1 and LPAR1 as intrinsic axon growth modulators for intact corticospinal motor neurons after adjacent injury. Furthermore, in vivo LPAR1 inhibition or LPPR1 overexpression enhances sprouting of intact corticospinal tract axons and yields greater functional recovery after unilateral brainstem lesion in wild-type mice. Thus, the transcriptional profile of injury-induced sprouting of intact neurons reveals targets for therapeutic enhancement of axon growth initiation and new synapse formation. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  5. NCBI nr-aa BLAST: CBRC-STRI-01-0474 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-0474 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8V...IC7|VN1A2_MOUSE RecName: Full=Vomeronasal type-1 receptor A2; Short=mV1R2; AltName: Full=Pheromone receptor ...2; AltName: Full=Vomeronasal type-1 receptor A9 dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasa...l 1 receptor, A2 [synthetic construct] gb|AAI40265.1| Vomeronasa...l 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal 1 receptor, A2 [Mus musculus] NP_035814.2 7e-81 53% ...

  6. NCBI nr-aa BLAST: CBRC-STRI-01-2574 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-2574 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8V...IC7|VN1A2_MOUSE RecName: Full=Vomeronasal type-1 receptor A2; Short=mV1R2; AltName: Full=Pheromone receptor ...2; AltName: Full=Vomeronasal type-1 receptor A9 dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasa...l 1 receptor, A2 [synthetic construct] gb|AAI40265.1| Vomeronasa...l 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal 1 receptor, A2 [Mus musculus] NP_035814.2 2e-85 52% ...

  7. NCBI nr-aa BLAST: CBRC-MMUS-06-0143 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0143 ref|NP_444448.1| vomeronasal 1 receptor, A3 [Mus musculus] sp|Q9EQ52|VN1A3_MOUSE Vomeronas...al type-1 receptor A3 (Vomeronasal type-1 receptor A6) gb|AAG42076.1|AF291482_1 vomeronasa...l receptor V1RA3 [Mus musculus] dbj|BAB79212.1| vomeronasal receptor 1 A6 [Mus musculus] gb|AAI41612.1| Vomeronasa...l 1 receptor, A3 [synthetic construct] gb|AAI40226.1| Vomeronasal 1 receptor, A3 [synthetic construct] NP_444448.1 4e-85 55% ...

  8. NCBI nr-aa BLAST: CBRC-STRI-01-0722 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-0722 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8V...IC7|VN1A2_MOUSE RecName: Full=Vomeronasal type-1 receptor A2; Short=mV1R2; AltName: Full=Pheromone receptor ...2; AltName: Full=Vomeronasal type-1 receptor A9 dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasa...l 1 receptor, A2 [synthetic construct] gb|AAI40265.1| Vomeronasa...l 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal 1 receptor, A2 [Mus musculus] NP_035814.2 7e-65 49% ...

  9. NCBI nr-aa BLAST: CBRC-STRI-01-1378 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-1378 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8V...IC7|VN1A2_MOUSE RecName: Full=Vomeronasal type-1 receptor A2; Short=mV1R2; AltName: Full=Pheromone receptor ...2; AltName: Full=Vomeronasal type-1 receptor A9 dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasa...l 1 receptor, A2 [synthetic construct] gb|AAI40265.1| Vomeronasa...l 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal 1 receptor, A2 [Mus musculus] NP_035814.2 4e-66 47% ...

  10. NCBI nr-aa BLAST: CBRC-MMUS-06-0144 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0144 ref|NP_444448.1| vomeronasal 1 receptor, A3 [Mus musculus] sp|Q9EQ52|VN1A3_MOUSE Vomeronas...al type-1 receptor A3 (Vomeronasal type-1 receptor A6) gb|AAG42076.1|AF291482_1 vomeronasa...l receptor V1RA3 [Mus musculus] dbj|BAB79212.1| vomeronasal receptor 1 A6 [Mus musculus] gb|AAI41612.1| Vomeronasa...l 1 receptor, A3 [synthetic construct] gb|AAI40226.1| Vomeronasal 1 receptor, A3 [synthetic construct] NP_444448.1 2e-66 52% ...

  11. NCBI nr-aa BLAST: CBRC-STRI-01-0309 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-0309 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8V...IC7|VN1A2_MOUSE RecName: Full=Vomeronasal type-1 receptor A2; Short=mV1R2; AltName: Full=Pheromone receptor ...2; AltName: Full=Vomeronasal type-1 receptor A9 dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasa...l 1 receptor, A2 [synthetic construct] gb|AAI40265.1| Vomeronasa...l 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal 1 receptor, A2 [Mus musculus] NP_035814.2 4e-81 54% ...

  12. NCBI nr-aa BLAST: CBRC-MMUS-06-0134 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0134 ref|NP_444448.1| vomeronasal 1 receptor, A3 [Mus musculus] sp|Q9EQ52|VN1A3_MOUSE Vomeronas...al type-1 receptor A3 (Vomeronasal type-1 receptor A6) gb|AAG42076.1|AF291482_1 vomeronasa...l receptor V1RA3 [Mus musculus] dbj|BAB79212.1| vomeronasal receptor 1 A6 [Mus musculus] gb|AAI41612.1| Vomeronasa...l 1 receptor, A3 [synthetic construct] gb|AAI40226.1| Vomeronasal 1 receptor, A3 [synthetic construct] NP_444448.1 1e-166 100% ...

  13. NCBI nr-aa BLAST: CBRC-STRI-01-1159 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-1159 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8V...IC7|VN1A2_MOUSE RecName: Full=Vomeronasal type-1 receptor A2; Short=mV1R2; AltName: Full=Pheromone receptor ...2; AltName: Full=Vomeronasal type-1 receptor A9 dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasa...l 1 receptor, A2 [synthetic construct] gb|AAI40265.1| Vomeronasa...l 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal 1 receptor, A2 [Mus musculus] NP_035814.2 4e-81 55% ...

  14. NCBI nr-aa BLAST: CBRC-STRI-01-1410 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-1410 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8V...IC7|VN1A2_MOUSE RecName: Full=Vomeronasal type-1 receptor A2; Short=mV1R2; AltName: Full=Pheromone receptor ...2; AltName: Full=Vomeronasal type-1 receptor A9 dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal... 1 receptor, A2 [synthetic construct] gb|AAI40265.1| Vomeronasal... 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal 1 receptor, A2 [Mus musculus] NP_035814.2 7e-87 53% ...

  15. NCBI nr-aa BLAST: CBRC-STRI-01-1107 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-1107 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8V...IC7|VN1A2_MOUSE RecName: Full=Vomeronasal type-1 receptor A2; Short=mV1R2; AltName: Full=Pheromone receptor ...2; AltName: Full=Vomeronasal type-1 receptor A9 dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal... 1 receptor, A2 [synthetic construct] gb|AAI40265.1| Vomeronasal... 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal 1 receptor, A2 [Mus musculus] NP_035814.2 2e-87 54% ...

  16. NCBI nr-aa BLAST: CBRC-STRI-01-2122 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-2122 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8V...IC7|VN1A2_MOUSE RecName: Full=Vomeronasal type-1 receptor A2; Short=mV1R2; AltName: Full=Pheromone receptor ...2; AltName: Full=Vomeronasal type-1 receptor A9 dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal... 1 receptor, A2 [synthetic construct] gb|AAI40265.1| Vomeronasal... 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal 1 receptor, A2 [Mus musculus] NP_035814.2 8e-88 54% ...

  17. NCBI nr-aa BLAST: CBRC-STRI-01-1216 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-1216 ref|NP_035814.2| vomeronasal 1 receptor, A2 [Mus musculus] sp|Q8V...IC7|VN1A2_MOUSE RecName: Full=Vomeronasal type-1 receptor A2; Short=mV1R2; AltName: Full=Pheromone receptor ...2; AltName: Full=Vomeronasal type-1 receptor A9 dbj|BAB79214.1| vomeronasal receptor 1 A9 [Mus musculus] gb|AAI41637.1| Vomeronasal... 1 receptor, A2 [synthetic construct] gb|AAI40265.1| Vomeronasal... 1 receptor, A2 [synthetic construct] gb|EDK99273.1| vomeronasal 1 receptor, A2 [Mus musculus] NP_035814.2 1e-71 50% ...

  18. NCBI nr-aa BLAST: CBRC-MMUS-06-0131 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-06-0131 ref|NP_444455.1| vomeronasal 1 receptor, B1 [Mus musculus] sp|Q9EP51|VN1B1_MOUSE Vomeronas...al type-1 receptor B1 (Vomeronasal type-1 receptor A5) (Vomeronasal receptor 2) (Ph...eromone receptor VN2) gb|AAG42083.1|AF291489_1 vomeronasal receptor V1RB1 [Mus musculus] gb|AAG43248.1| VN2 ...[Mus musculus] gb|AAI07184.1| Vomeronasal 1 receptor, B1 [Mus musculus] gb|EDK99276.1| vomeronasal 1 receptor, B1 [Mus musculus] NP_444455.1 1e-133 76% ...

  19. Colonization, mouse-style

    Directory of Open Access Journals (Sweden)

    Searle Jeremy B

    2010-10-01

    Full Text Available Abstract Several recent papers, including one in BMC Evolutionary Biology, examine the colonization history of house mice. As well as background for the analysis of mouse adaptation, such studies offer a perspective on the history of movements of the humans that accidentally transported the mice. See research article: http://www.biomedcentral.com/1471-2148/10/325

  20. Mouse Leydig Tumor Cells

    Directory of Open Access Journals (Sweden)

    Bo-Syong Pan

    2011-01-01

    Full Text Available Cordycepin is a natural pure compound extracted from Cordyceps sinensis (CS. We have demonstrated that CS stimulates steroidogenesis in primary mouse Leydig cell and activates apoptosis in MA-10 mouse Leydig tumor cells. It is highly possible that cordycepin is the main component in CS modulating Leydig cell functions. Thus, our aim was to investigate the steroidogenic and apoptotic effects with potential mechanism of cordycepin on MA-10 mouse Leydig tumor cells. Results showed that cordycepin significantly stimulated progesterone production in dose- and time-dependent manners. Adenosine receptor (AR subtype agonists were further used to treat MA-10 cells, showing that A1, A 2A , A 2B , and A3, AR agonists could stimulate progesterone production. However, StAR promoter activity and protein expression remained of no difference among all cordycepin treatments, suggesting that cordycepin might activate AR, but not stimulated StAR protein to regulate MA-10 cell steroidogenesis. Meanwhile, cordycepin could also induce apoptotic cell death in MA-10 cells. Moreover, four AR subtype agonists induced cell death in a dose-dependent manner, and four AR subtype antagonists could all rescue cell death under cordycepin treatment in MA-10 cells. In conclusion, cordycepin could activate adenosine subtype receptors and simultaneously induce steroidogenesis and apoptosis in MA-10 mouse Leydig tumor cells.

  1. The Mouse SAGE Site: database of public mouse SAGE libraries.

    Science.gov (United States)

    Divina, Petr; Forejt, Jirí

    2004-01-01

    The Mouse SAGE Site is a web-based database of all available public libraries generated by the Serial Analysis of Gene Expression (SAGE) from various mouse tissues and cell lines. The database contains mouse SAGE libraries organized in a uniform way and provides web-based tools for browsing, comparing and searching SAGE data with reliable tag-to-gene identification. A modified approach based on the SAGEmap database is used for reliable tag identification. The Mouse SAGE Site is maintained on an ongoing basis at the Institute of Molecular Genetics, Academy of Sciences of the Czech Republic and is accessible at the internet address http://mouse.biomed.cas.cz/sage/.

  2. Impact of local endothelial challenge with cytomegalovirus or glycoprotein B on vasodilation in intact pressurized arteries from nonpregnant and pregnant mice.

    Science.gov (United States)

    Gombos, Randi B; Teefy, Jana; Lee, Albert; Hemmings, Denise G

    2012-10-01

    Cytomegalovirus (CMV) infections are associated with vascular diseases in the human population. We have previously shown vascular dysfunction in systemic and uterine arteries dissected from nonpregnant (NP) mouse CMV (mCMV)-infected mice that was further impaired during late pregnancy (LP). CMV attachment alone through glycoprotein B (GB) can generate signals that impact vascular tone regulation. However, the contribution of direct virus interactions with endothelium to the vascular dysfunction we previously observed after in vivo mCMV infection is not known. We used a pressure myograph system to infuse GB or whole intact mCMV inside arteries dissected from uninfected mice and assessed vasodilation to methacholine infused inside pressurized arteries rather than applied abluminally. These results were compared to those observed after methacholine infusion into untreated arteries dissected from mCMV-infected mice. In mesenteric arteries, vasodilation to infused methacholine did not differ among treatments in NP or LP groups in contrast to previously published studies. However, increased vasoconstrictor activity was unmasked after blocking thromboxane receptors or prostaglandin production. Vasodilation in uterine arteries from uninfected NP mice to infused methacholine was increased by both GB and whole intact mCMV pretreatment. Untreated uterine arteries from mCMV-infected NP mice showed even greater vasodilation. There was no effect of GB or whole intact mCMV pretreatment in uterine arteries from uninfected LP mice, whereas vasodilation to infused methacholine was reduced in untreated uterine arteries from mCMV-infected LP mice. CMV exerts direct effects on vascular function which should be considered during viral reactivation leading to viremia and during GB-based vaccine administration.

  3. Performance of intact and castrated beef cattle in an intensive croppasture rotation system

    Directory of Open Access Journals (Sweden)

    Tercilio Turini

    2015-07-01

    Full Text Available This research had as objective to evaluate the performance of intact or castrated beef cattle in a croppasture rotation system. The experiment was conducted during 2004 and 2005, and carried out at the Cooperativa Agropecuária Mourãoense (COAMO Experimental Farm, in Campo Mourão city, Paraná state. It was used a completely randomized design, with two treatments, intact or castrated. Forty ½Angus+½Nelore crossbred animals, with average age of nine months, were used. Half of the animals were castrated at weaning, and the other half was kept intact. Pasture was composed of two areas. The winter field, established after soybean crop, was composed by a mixture of black oat (Avena strigosa and Italian ryegrass (Lolium multiforum. The summer field was composed by stargrass (Cynodon nlemfuensis and Mombaça grass (Panicum maximum. During the winter time it was used a continues grazing system, with regulator animals (put and take, and during the summer an intensive rotational system, with regulator animals and fixed grazing period. Intact animals presented higher average daily weight gain (0.907 vs 0.698 kg, slaughter weight (490.9 vs 442.2 kg, and hot carcass weight (250.2 vs 232.6 kg. Slaughter age was influenced by sexual condition, being lesser in the intact animals. Carcass dressing percentage was similar for the groups. Castrated animals showed better finishing fat cover and backfat thickness (3.45 vs 2.70 mm compared to intact ones. Therefore, it can be concluded that intact animals presents better performance than castrated ones when finished in an intensive crop-pasture rotation system, however, they may not present the minimum required fat cover, when slaughter at young ages.

  4. Comparison of ripening processes in intact tomato fruit and excised pericarp discs.

    Science.gov (United States)

    Campbell, A D; Huysamer, M; Stotz, H U; Greve, L C; Labavitch, J M

    1990-12-01

    Physiological processes characteristic of ripening in tissues of intact tomato fruit (Lycopersicon esculentum Mill.) were examined in excised pericarp discs. Pericarp discs were prepared from mature-green tomato fruit and stored in 24-well culture plates, in which individual discs could be monitored for color change, ethylene biosynthesis, and respiration, and selected for cell wall analysis. Within the context of these preparation and handling procedures, most whole fruit ripening processes were maintained in pericarp discs. Pericarp discs and matched intact fruit passed through the same skin color stages at similar rates, as expressed in the L(*)a(*)b(*) color space, changing from green (a(*) red (a(*) > 15) in about 6 days. Individual tissues of the pericarp discs changed color in the same sequence seen in intact fruit (exocarp, endocarp, then vascular parenchyma). Discs from different areas changed in the same spatial sequence seen in intact fruit (bottom, middle, top). Pericarp discs exhibited climacteric increases in ethylene biosynthesis and CO(2) production comparable with those seen in intact fruit, but these were more tightly linked to rate of color change, reaching a peak around a(*) = 5. Tomato pericarp discs decreased in firmness as color changed. Cell wall carbohydrate composition changed with color as in intact fruit: the quantity of water-soluble pectin eluted from the starch-free alcohol insoluble substances steadily increased and more tightly bound, water-insoluble, pectin decreased in inverse relationship. The cell wall content of the neutral sugars arabinose, rhamnose, and galactose steadily decreased as color changed. The extractable activity of specific cell wall hydrolases changed as in intact fruit: polygalacturonase activity, not detectable in green discs (a(*) = -5), appeared as discs turned yellow-red (a(*) = 5), and increased another eight-fold as discs became full red (a(*) value +20). Carboxymethyl-cellulase activity, low in extracts

  5. Human and mouse tissue-engineered small intestine both demonstrate digestive and absorptive function.

    Science.gov (United States)

    Grant, Christa N; Mojica, Salvador Garcia; Sala, Frederic G; Hill, J Ryan; Levin, Daniel E; Speer, Allison L; Barthel, Erik R; Shimada, Hiroyuki; Zachos, Nicholas C; Grikscheit, Tracy C

    2015-04-15

    Short bowel syndrome (SBS) is a devastating condition in which insufficient small intestinal surface area results in malnutrition and dependence on intravenous parenteral nutrition. There is an increasing incidence of SBS, particularly in premature babies and newborns with congenital intestinal anomalies. Tissue-engineered small intestine (TESI) offers a therapeutic alternative to the current standard treatment, intestinal transplantation, and has the potential to solve its biggest challenges, namely donor shortage and life-long immunosuppression. We have previously demonstrated that TESI can be generated from mouse and human small intestine and histologically replicates key components of native intestine. We hypothesized that TESI also recapitulates native small intestine function. Organoid units were generated from mouse or human donor intestine and implanted into genetically identical or immunodeficient host mice. After 4 wk, TESI was harvested and either fixed and paraffin embedded or immediately subjected to assays to illustrate function. We demonstrated that both mouse and human tissue-engineered small intestine grew into an appropriately polarized sphere of intact epithelium facing a lumen, contiguous with supporting mesenchyme, muscle, and stem/progenitor cells. The epithelium demonstrated major ultrastructural components, including tight junctions and microvilli, transporters, and functional brush-border and digestive enzymes. This study demonstrates that tissue-engineered small intestine possesses a well-differentiated epithelium with intact ion transporters/channels, functional brush-border enzymes, and similar ultrastructural components to native tissue, including progenitor cells, whether derived from mouse or human cells.

  6. Parasite load in intact and ulcerative skin of dogs with leishmaniais

    Directory of Open Access Journals (Sweden)

    Francine Maria de França Silva

    2016-03-01

    Full Text Available Abstract The skin is the site of inoculation of Leishmania spp. in susceptible hosts, and consequently dermatopathies, especially ulcerative dermatitis, are the main clinical signs observed. The aim of this study was to assess parasitism of the skin (intact and ulcerated among dogs that were naturally infected by Leishmania spp., through immunohistochemical analysis. Skin fragments (intact and ulcerated were collected from 13 dogs with positive parasitological (bone marrow aspiration and exfoliative skin and serological examinations (ELISA S7® Biogene forLeishmania spp. These samples were processed using the immunohistochemical technique, involving the streptavidin-peroxidase complex. Ulcerative lesions were mainly observed on the elbows (53.84%; 7/13, nostrils (15.38%; 2/13, ears (23.07%; 3/13 and wings of the ilium (7.69%; 1/13. A severe parasite load was detected in 46.15% and 76.92% of the intact and ulcerated skin samples tested, respectively. The parasite load on ulcerated skin was statistically higher than on intact skin (p = 0.0221. These results indicate that the intact and ulcerated skin may host a high parasite load of amastigote forms of Leishmania spp., which can favor the transmission of the parasite.

  7. Apoplastic superoxide production and peroxidase activity by intact and excised axenically grown seedling roots of sunflower.

    Science.gov (United States)

    Garrido, Inmaculada; Espinosa, Francisco; Alvarez-Tinaut, M Carmen

    2012-10-01

    Excised and cold-preincubated sunflower seedling roots were compared with intact non-preincubated roots to test the effect of the injury stress and cold preincubation on the oxidative burst measured as apoplastic superoxide (O (2) (.-) ) generation and exocellular peroxidase (ECPOX) activity. Preincubated excised or intact roots released into the medium apoplastic proteins with peroxidase activity. Intact and excised roots responded to methyl jasmonate by an immediate oxidative burst that could not be induced by salicylic acid; both phytohormones also induced a slight and slow O (2)(.-) generation and ECPOX activity on excised roots, when added to the cold preincubation medium. The results with cyanide, azide, SHAM (ECPOX inhibitors) and diphenylene iodonium (inhibitor of trans-plasma membrane NAD(P)H-oxidases (NOX)-respiratory burst oxidase homologue in plants (RBOH), the trans-plasmamembrane nicotinamide adenine dinucleotide phosphate oxidase) are consistent with the hypothesis that different systems may be the origin of O (2) (.-) in intact and excised roots; ECPOX was an important component of them in both, together with NOX-RBOH in intact roots, but in excised roots the last one was replaced by an oxidase sensitive to the same inhibitors as the alternative mitochondrial oxidase. According to our hypothesis, these results could be explained if the electron flux would be deviated to different interconnected plasma membrane-redox systems, with different terminal oxidases, activated by different effectors or stresses.

  8. Near-infrared reflectance spectroscopy for predicting amino acids content in intact processed animal proteins.

    Science.gov (United States)

    De la Haba, Maria José; Garrido-Varo, Ana; Guerrero-Ginel, José Emilio; Pérez-Marín, Dolores C

    2006-10-01

    Near-infrared calibrations were developed for the instantaneous prediction of amino acids composition of processed animal proteins (PAPs). Two sample presentation modes were compared (ground vs intact) for demonstrating the viability of the analysis in the intact form, avoiding the need for milling. Modified partial least-squares (MPLS) equations for the prediction of amino acids in PAPs were developed using the same set of samples (N = 92 PAPs) analyzed in ground and intact form and in three cups differing in the optical window size. The standard error for cross validation (SECV) and the coefficient of determination (1-VR) values yielded with the calibrations developed using the samples analyzed in the intact form showed similar or even better accuracy than those obtained with finely ground samples. The excellent predictive ability (1-VR > 0.90; CV < 3.0%) obtained for the prediction of amino acids in intact processed animal proteins opens an enormous expectative for the on-line implementation of NIRS technology in the processing and marketing of these important protein feed ingredients, alleviating the costs and time associated with the routine quality controls.

  9. Transient increase of intact visual field size by high-frequency narrow-band stimulation.

    Science.gov (United States)

    Elliott, Mark A; Seifert, Doerthe; Poggel, Dorothe A; Strasburger, Hans

    2015-03-01

    Three patients with visual field defects were stimulated with a square matrix pattern, either static, or flickering at frequencies that had been found to either promote or not promote blindsight performance. Comparison between pre- and post-stimulation perimetric maps revealed an increase in the size of the intact visual field but only for flicker frequencies previously found to promote blindsight. These changes were temporary but dramatic - in two instances the intact field was increased by an area of ∼30 deg(2) of visual angle. These results indicate that not only does specific high-frequency stimulus flicker promote blindsight, but that intact visual field size may be increased by stimulation at the same frequencies. Our findings inform speculation on both the brain mechanisms and the potency of temporal modulation for altering the functional visual field.

  10. Spontaneous and light-induced photon emission from intact brains of chick embryos

    Institute of Scientific and Technical Information of China (English)

    张锦珠; 于文斗; 孙彤

    1997-01-01

    Photon emission (PE) and light-induced photon emission(LPE) of intact brains isolated from chick embryos have been measured by using the single photon counting device. Experimental results showed that the intensi-ty level of photon emission was detected to be higher from intact brain than from the medium in which the brain was immerged during measuring, and the emission intensity was related to the developmental stages, the healthy situation of the measured embryos, and the freshness of isolated brains as well. After white light illumination, a short-life de-layed emission from intact brains was observed, and its relaxation behavior followed a hyperbolic rather than an expo-nential law. According to the hypothesis of biophoton emission originating from a delocalized coherent electromagnetic field and Frohlich’s idea of coherent long-range interactions in biological systems, discussions were made on the signifi-cance of photon emission in studying cell communication, biological regulation, living system’

  11. Isolation of Mouse Neutrophils.

    Science.gov (United States)

    Swamydas, Muthulekha; Luo, Yi; Dorf, Martin E; Lionakis, Michail S

    2015-08-03

    Neutrophils represent the first line of defense against bacterial and fungal pathogens. Indeed, patients with inherited and acquired qualitative and quantitative neutrophil defects are at high risk for developing bacterial and fungal infections and suffering adverse outcomes from these infections. Therefore, research aiming at defining the molecular factors that modulate neutrophil effector function under homeostatic conditions and during infection is essential for devising strategies to augment neutrophil function and improve the outcome of infected individuals. This unit describes a reproducible density gradient centrifugation-based protocol that can be applied in any laboratory to harvest large numbers of highly enriched and highly viable neutrophils from the bone marrow of mice both at the steady state and following infection with Candida albicans as described in UNIT. In another protocol, we also present a method that combines gentle enzymatic tissue digestion with a positive immunomagnetic selection technique or Fluorescence-activated cell sorting (FACS) to harvest highly pure and highly viable preparations of neutrophils directly from mouse tissues such as the kidney, the liver or the spleen. Finally, methods for isolating neutrophils from mouse peritoneal fluid and peripheral blood are included. Mouse neutrophils isolated by these protocols can be used for examining several aspects of cellular function ex vivo including pathogen binding, phagocytosis and killing, neutrophil chemotaxis, oxidative burst, degranulation and cytokine production, and for performing neutrophil adoptive transfer experiments.

  12. RIKEN mouse genome encyclopedia.

    Science.gov (United States)

    Hayashizaki, Yoshihide

    2003-01-01

    We have been working to establish the comprehensive mouse full-length cDNA collection and sequence database to cover as many genes as we can, named Riken mouse genome encyclopedia. Recently we are constructing higher-level annotation (Functional ANnoTation Of Mouse cDNA; FANTOM) not only with homology search based annotation but also with expression data profile, mapping information and protein-protein database. More than 1,000,000 clones prepared from 163 tissues were end-sequenced to classify into 159,789 clusters and 60,770 representative clones were fully sequenced. As a conclusion, the 60,770 sequences contained 33,409 unique. The next generation of life science is clearly based on all of the genome information and resources. Based on our cDNA clones we developed the additional system to explore gene function. We developed cDNA microarray system to print all of these cDNA clones, protein-protein interaction screening system, protein-DNA interaction screening system and so on. The integrated database of all the information is very useful not only for analysis of gene transcriptional network and for the connection of gene to phenotype to facilitate positional candidate approach. In this talk, the prospect of the application of these genome resourced should be discussed. More information is available at the web page: http://genome.gsc.riken.go.jp/.

  13. Mouse models in oncoimmunology.

    Science.gov (United States)

    Zitvogel, Laurence; Pitt, Jonathan M; Daillère, Romain; Smyth, Mark J; Kroemer, Guido

    2016-12-01

    Fundamental cancer research and the development of efficacious antineoplastic treatments both rely on experimental systems in which the relationship between malignant cells and immune cells can be studied. Mouse models of transplantable, carcinogen-induced or genetically engineered malignancies - each with their specific advantages and difficulties - have laid the foundations of oncoimmunology. These models have guided the immunosurveillance theory that postulates that evasion from immune control is an essential feature of cancer, the concept that the long-term effects of conventional cancer treatments mostly rely on the reinstatement of anticancer immune responses and the preclinical development of immunotherapies, including currently approved immune checkpoint blockers. Specific aspects of pharmacological development, as well as attempts to personalize cancer treatments using patient-derived xenografts, require the development of mouse models in which murine genes and cells are replaced with their human equivalents. Such 'humanized' mouse models are being progressively refined to characterize the leukocyte subpopulations that belong to the innate and acquired arms of the immune system as they infiltrate human cancers that are subjected to experimental therapies. We surmise that the ever-advancing refinement of murine preclinical models will accelerate the pace of therapeutic optimization in patients.

  14. Neural activation differences in amputees during imitation of intact versus amputee movements

    Directory of Open Access Journals (Sweden)

    William F Cusack

    2012-06-01

    Full Text Available The mirror neuron system has been attributed with increased activation in motor-related cortical areas upon viewing of another’s actions. Recent work suggests that limb movements that are similar and dissimilar in appearance to that of the viewer equivalently activate the mirror neuron system. It is unclear if this result can be observed in the action encoding areas in amputees who use prosthetic devices. Intact subjects and upper extremity amputee prosthesis users were recruited to view video demonstrations of tools being used by an intact actor and a prosthetic device user. All subjects were asked to pantomime the movements seen in the video while recording electroencephalography. Intact subjects showed equivalent left parietofrontal activity during imitation after watching the intact or prosthetic arm. Likewise, when prosthesis users imitated prosthesis demonstrations, typical left parietofrontal activation was observed during planning. When prosthesis users imitated intact actors, a new pattern was revealed which showed greater bilateral parietal and occipital activity during movement planning (p<0.001. This change may be required for prosthesis users to imitate movements in which the limb states between the observed and the observer do not match. The finding that prosthesis users imitating other prosthesis users showed typical left parietofrontal activation suggests that these subjects engage normal planning related activity when they are able to imitate a limb matching their own. This result has significant implications on rehabilitation, as standard therapy involves training with an intact occupational therapist, which could necessitate atypical planning mechanisms in amputees when learning to use their prosthesis.

  15. Effect of tail suspension on haemodynamics in intact and sympathectomized rats.

    Science.gov (United States)

    Tarasova, O; Figourina, I; Zotov, A; Borovik, A; Vinogradova, O

    2001-09-01

    The effect of a 2-week tail suspension on mean arterial pressure (MAP) and pulse interval (PI) was studied in conscious chronically instrumented intact and sympathectomized rats. Sympathectomy was induced by 6-hydroxydopamine (100 mg x kg(-1), i.v.), injected 1 day before, and on days 5 and 10 of suspension. During suspension the intact rats had the same levels of MAP and PI as the intact controls. After release from suspension their MAP did not change: PI decreased (by 9%), but only for 1 h. Sympathectomy augmented the haemodynamic changes after release from suspension: tachycardia (10%) and hypotension (19%) were observed during the entire recording period (2 h). Therefore, in rats, post-suspension hypotension becomes apparent only after elimination of sympathetic influences. In spite of unaltered systemic parameters, intact post-suspension rats showed diminished responses to intravenous administration of tyramine (100 microg x kg(-1)) and phenylephrine (1-2 microg x kg(-1)). In addition, they showed augmented haemodynamic changes associated with natural behaviour. The increase of MAP and the decrease of PI on transition from rest to movement were more pronounced in the intact post-suspension rats than in the control rats (MAP 8.3% vs 4.5%, PI 15% vs 9%). In control rats the spectrum power density of low-frequency (0.0195-0.25 Hz) MAP fluctuations depended only weakly on behavioural activity, whereas in post-suspension intact rats it was profoundly augmented during movements (by 170%). Since the rapid adjustment of haemodynamics to behavioural activity is controlled by the sympathetic nervous system, the unbalanced condition of the cardiovascular system after suspension may reflect an altered sympathetic nerve control of the circulation.

  16. Losing a jewel—Rapid declines in Myanmar’s intact forests from 2002-2014

    Science.gov (United States)

    Horning, Ned; Khaing, Thiri; Thein, Zaw Min; Aung, Kyaw Moe; Aung, Kyaw Htet; Phyo, Paing; Tun, Ye Lin; Oo, Aung Htat; Neil, Anthony; Thu, Win Myo; Songer, Melissa; Huang, Qiongyu; Connette, Grant; Leimgruber, Peter

    2017-01-01

    New and rapid political and economic changes in Myanmar are increasing the pressures on the country’s forests. Yet, little is known about the past and current condition of these forests and how fast they are declining. We mapped forest cover in Myanmar through a consortium of international organizations and environmental non-governmental groups, using freely-available public domain data and open source software tools. We used Landsat satellite imagery to assess the condition and spatial distribution of Myanmar’s intact and degraded forests with special focus on changes in intact forest between 2002 and 2014. We found that forests cover 42,365,729 ha or 63% of Myanmar, making it one of the most forested countries in the region. However, severe logging, expanding plantations, and degradation pose increasing threats. Only 38% of the country’s forests can be considered intact with canopy cover >80%. Between 2002 and 2014, intact forests declined at a rate of 0.94% annually, totaling more than 2 million ha forest loss. Losses can be extremely high locally and we identified 9 townships as forest conversion hotspots. We also delineated 13 large (>100,000 ha) and contiguous intact forest landscapes, which are dispersed across Myanmar. The Northern Forest Complex supports four of these landscapes, totaling over 6.1 million ha of intact forest, followed by the Southern Forest Complex with three landscapes, comprising 1.5 million ha. These remaining contiguous forest landscape should have high priority for protection. Our project demonstrates how open source data and software can be used to develop and share critical information on forests when such data are not readily available elsewhere. We provide all data, code, and outputs freely via the internet at (for scripts: https://bitbucket.org/rsbiodiv/; for the data: http://geonode.themimu.info/layers/geonode%3Amyan_lvl2_smoothed_dec2015_resamp) PMID:28520726

  17. Quantitative Analysis of Human Salivary Gland-Derived Intact Proteome Using Top-Down Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Si; Brown, Joseph N.; Tolic, Nikola; Meng, Da; Liu, Xiaowen; Zhang, Haizhen; Zhao, Rui; Moore, Ronald J.; Pevzner, Pavel A.; Smith, Richard D.; Pasa-Tolic, Ljiljana

    2014-05-31

    There are several notable challenges inherent to fully characterizing the entirety of the human saliva proteome using bottom-up approaches, including polymorphic isoforms, post-translational modifications, unique splice variants, deletions, and truncations. To address these challenges, we have developed a top-down based liquid chromatography-mass spectrometry (LC-MS) approach, which cataloged 20 major human salivary proteins with a total of 83 proteoforms, containing a broad range of post-translational modifications. Among these proteins, several previously reported disease biomarker proteins were identified at the intact protein level, such as beta-2 microglobulin (B2M). In addition, intact glycosylated proteoforms of several saliva proteins were also characterized, including intact N-glycosylated protein prolactin inducible protein (PIP) and O-glycosylated acidic protein rich protein (aPRP). These characterized proteoforms constitute an intact saliva proteoform database, which was used for quantitative comparison of intact salivary proteoforms among six healthy individuals. Human parotid (PS) and submandibular/sublingual gland (SMSL) secretion samples (2 μg of protein each) from six healthy individuals were compared using RPLC coupled with the 12T FTICR mass spectrometer. Significantly different protein and PTM patterns were resolved with high reproducibility between PS and SMSL glands. The results from this study provide further insight into the potential mechanisms of PTM pathways in oral glandular secretion, expanding our knowledge of this complex yet easily accessible fluid. Intact protein LC-MS approach presented herein can potentially be applied for rapid and accurate identification of biomarkers from only a few microliters of human glandular saliva.

  18. Multivariate Calibration Approach for Quantitative Determination of Cell-Line Cross Contamination by Intact Cell Mass Spectrometry and Artificial Neural Networks.

    Directory of Open Access Journals (Sweden)

    Elisa Valletta

    Full Text Available Cross-contamination of eukaryotic cell lines used in biomedical research represents a highly relevant problem. Analysis of repetitive DNA sequences, such as Short Tandem Repeats (STR, or Simple Sequence Repeats (SSR, is a widely accepted, simple, and commercially available technique to authenticate cell lines. However, it provides only qualitative information that depends on the extent of reference databases for interpretation. In this work, we developed and validated a rapid and routinely applicable method for evaluation of cell culture cross-contamination levels based on mass spectrometric fingerprints of intact mammalian cells coupled with artificial neural networks (ANNs. We used human embryonic stem cells (hESCs contaminated by either mouse embryonic stem cells (mESCs or mouse embryonic fibroblasts (MEFs as a model. We determined the contamination level using a mass spectra database of known calibration mixtures that served as training input for an ANN. The ANN was then capable of correct quantification of the level of contamination of hESCs by mESCs or MEFs. We demonstrate that MS analysis, when linked to proper mathematical instruments, is a tangible tool for unraveling and quantifying heterogeneity in cell cultures. The analysis is applicable in routine scenarios for cell authentication and/or cell phenotyping in general.

  19. Nonspecific airway reactivity in a mouse model of asthma

    Energy Technology Data Exchange (ETDEWEB)

    Collie, D.D.; Wilder, J.A.; Bice, D.E.

    1995-12-01

    Animal models are indispensable for studies requiring an intact immune system, especially for studying the pathogenic mechanisms in atopic diseases, regulation of IgE production, and related biologic effects. Mice are particularly suitable and have been used extensively for such studies because their immune system is well characterized. Further, large numbers of mutants or inbred strains of mice are available that express deficiencies of individual immunologic processes, inflammatory cells, or mediator systems. By comparing reactions in such mice with appropriate control animals, the unique roles of individual cells or mediators may be characterized more precisely in the pathogenesis of atopic respiratory diseases including asthma. However, given that asthma in humans is characterized by the presence of airway hyperresponsiveness to specific and nonspecific stimuli, it is important that animal models of this disease exhibit similar physiologic abnormalities. In the past, the size of the mouse has limited its versatility in this regard. However, recent studies indicate the feasibility of measuring pulmonary responses in living mice, thus facilitating the physiologic evaluation of putative mouse models of human asthma that have been well charcterized at the immunologic and patholigic level. Future work will provide details of the morphometry of the methacholine-induced bronchoconstriction and will further seek to determine the relationship between cigarette smoke exposure and the development of NS-AHR in the transgenic mouse model.

  20. Imaging of functional connectivity in the mouse brain.

    Science.gov (United States)

    White, Brian R; Bauer, Adam Q; Snyder, Abraham Z; Schlaggar, Bradley L; Lee, Jin-Moo; Culver, Joseph P

    2011-01-20

    Functional neuroimaging (e.g., with fMRI) has been difficult to perform in mice, making it challenging to translate between human fMRI studies and molecular and genetic mechanisms. A method to easily perform large-scale functional neuroimaging in mice would enable the discovery of functional correlates of genetic manipulations and bridge with mouse models of disease. To satisfy this need, we combined resting-state functional connectivity mapping with optical intrinsic signal imaging (fcOIS). We demonstrate functional connectivity in mice through highly detailed fcOIS mapping of resting-state networks across most of the cerebral cortex. Synthesis of multiple network connectivity patterns through iterative parcellation and clustering provides a comprehensive map of the functional neuroarchitecture and demonstrates identification of the major functional regions of the mouse cerebral cortex. The method relies on simple and relatively inexpensive camera-based equipment, does not require exogenous contrast agents and involves only reflection of the scalp (the skull remains intact) making it minimally invasive. In principle, fcOIS allows new paradigms linking human neuroscience with the power of molecular/genetic manipulations in mouse models.

  1. Imaging of functional connectivity in the mouse brain.

    Directory of Open Access Journals (Sweden)

    Brian R White

    Full Text Available Functional neuroimaging (e.g., with fMRI has been difficult to perform in mice, making it challenging to translate between human fMRI studies and molecular and genetic mechanisms. A method to easily perform large-scale functional neuroimaging in mice would enable the discovery of functional correlates of genetic manipulations and bridge with mouse models of disease. To satisfy this need, we combined resting-state functional connectivity mapping with optical intrinsic signal imaging (fcOIS. We demonstrate functional connectivity in mice through highly detailed fcOIS mapping of resting-state networks across most of the cerebral cortex. Synthesis of multiple network connectivity patterns through iterative parcellation and clustering provides a comprehensive map of the functional neuroarchitecture and demonstrates identification of the major functional regions of the mouse cerebral cortex. The method relies on simple and relatively inexpensive camera-based equipment, does not require exogenous contrast agents and involves only reflection of the scalp (the skull remains intact making it minimally invasive. In principle, fcOIS allows new paradigms linking human neuroscience with the power of molecular/genetic manipulations in mouse models.

  2. [A histochemical study of acetylcholinesterase in intact and deafferented cat auditory cortex].

    Science.gov (United States)

    Genis, E D

    1976-01-01

    The peculiarities of the AChE distribution were investigated in the intact cat auditory cortex and during early period of its neuronal isolation. It is shown that in the isolated cortex slab the staining of the AChE containing fibre disappeared from the neuropile, while in the intact cortex it was well pronounced. AChE accumulation was observed in the proximal parts of the transsected thalamo-cortical fibres. It is supposed that the AChE-containing fibres in the auditory cortex belong to nonspecific thalamic inputs.

  3. Population Studies of Intact Vitamin D Binding Protein by Affinity Capture ESI-TOF-MS

    Science.gov (United States)

    Borges, Chad R.; Jarvis, Jason W.; Oran, Paul E.; Rogers, Stephen P.; Nelson, Randall W.

    2008-01-01

    Blood plasma proteins with molecular weights greater than approximately 30 kDa are refractory to comprehensive, high-throughput qualitative characterization of microheterogeneity across human populations. Analytical techniques for obtaining high mass resolution for targeted, intact protein characterization and, separately, high sample throughput exist, but efficient means of coupling these assay characteristics remain rather limited. This article discusses the impetus for analyzing intact proteins in a targeted manner across populations and describes the methodology required to couple mass spectrometric immunoassay with electrospray ionization mass spectrometry for the purpose of qualitatively characterizing a prototypical large plasma protein, vitamin D binding protein, across populations. PMID:19137103

  4. Relationship between parental aspiration and academic achievement of Xhosa children from broken and intact families.

    Science.gov (United States)

    Cherian, V I

    1994-06-01

    This study concerned the relationship between parental aspiration and academic achievement of Xhosa pupils (369 boys and 652 girls) whose ages ranged from 13 to 17 years (mean age, 15.3 yr.). Children were chosen at random from the Standard 7/Year 9 population of Transkei. A questionnaire administered to parents or parent surrogates identified 242 children of parents divorced or separated and 713 from intact homes and obtained parental aspiration for the education of children. Analysis of variance showed significant effects of parental aspiration on academic achievement of children whether the children were from broken or intact homes.

  5. Assessment of contractility in intact ventricular cardiomyocytes using the dimensionless 'Frank-Starling Gain' index.

    Science.gov (United States)

    Bollensdorff, Christian; Lookin, Oleg; Kohl, Peter

    2011-07-01

    This paper briefly recapitulates the Frank-Starling law of the heart, reviews approaches to establishing diastolic and systolic force-length behaviour in intact isolated cardiomyocytes, and introduces a dimensionless index called 'Frank-Starling Gain', calculated as the ratio of slopes of end-systolic and end-diastolic force-length relations. The benefits and limitations of this index are illustrated on the example of regional differences in Guinea pig intact ventricular cardiomyocyte mechanics. Potential applicability of the Frank-Starling Gain for the comparison of cell contractility changes upon stretch will be discussed in the context of intra- and inter-individual variability of cardiomyocyte properties.

  6. The impact of family intactness on family functioning, parental control and parent-child relational qualities in a Chinese context

    Directory of Open Access Journals (Sweden)

    Daniel Tan Lei Shek

    2015-01-01

    Full Text Available The current study investigated the differences between intact and non-intact families in family processes, including systematic family functioning, parental behavioral control, parental psychological control, and parent-child relational qualities. The participants were 3,328 Secondary One students, with a mean age of 12.59 years, recruited from 28 secondary schools in Hong Kong. Four validated scales were used to assess family processes. Results showed that adolescents in non-intact families perceived relatively poorer family functioning, lower level of paternal and maternal behavioral control, lower level of paternal psychological control and poorer parent-child relational qualities than did adolescents in intact families. This generally indicated that family processes were poorer in non-intact families, compared with those in intact families. The theoretical and practical implications of the findings were discussed.

  7. Mouse genetics: Catalogue and scissors

    Directory of Open Access Journals (Sweden)

    Han-Woong Lee

    2012-12-01

    Full Text Available Phenotypic analysis of gene-specific knockout (KO mice hasrevolutionized our understanding of in vivo gene functions. Asthe use of mouse embryonic stem (ES cells is inevitable forconventional gene targeting, the generation of knockout miceremains a very time-consuming and expensive process. Toaccelerate the large-scale production and phenotype analyses ofKO mice, international efforts have organized global consortiasuch as the International Knockout Mouse Consortium (IKMCand International Mouse Phenotype Consortium (IMPC, andthey are persistently expanding the KO mouse catalogue that ispublicly available for the researches studying specific genes ofinterests in vivo. However, new technologies, adoptingzinc-finger nucleases (ZFNs or Transcription Activator-LikeEffector (TALE Nucleases (TALENs to edit the mouse genome,are now emerging as valuable and effective shortcuts alternativefor the conventional gene targeting using ES cells. Here, weintroduce the recent achievement of IKMC, and evaluate thesignificance of ZFN/TALEN technology in mouse genetics.

  8. Mouse genetics: catalogue and scissors.

    Science.gov (United States)

    Sung, Young Hoon; Baek, In-Jeoung; Seong, Je Kyung; Kim, Jin Soo; Lee, Han-Woong

    2012-12-01

    Phenotypic analysis of gene-specific knockout (KO) mice has revolutionized our understanding of in vivo gene functions. As the use of mouse embryonic stem (ES) cells is inevitable for conventional gene targeting, the generation of knockout mice remains a very time-consuming and expensive process. To accelerate the large-scale production and phenotype analyses of KO mice, international efforts have organized global consortia such as the International Knockout Mouse Consortium (IKMC) and International Mouse Phenotype Consortium (IMPC), and they are persistently expanding the KO mouse catalogue that is publicly available for the researches studying specific genes of interests in vivo. However, new technologies, adopting zinc-finger nucleases (ZFNs) or Transcription Activator-Like Effector (TALE) Nucleases (TALENs) to edit the mouse genome, are now emerging as valuable and effective shortcuts alternative for the conventional gene targeting using ES cells. Here, we introduce the recent achievement of IKMC, and evaluate the significance of ZFN/TALEN technology in mouse genetics.

  9. Human anti-mouse antibodies.

    Science.gov (United States)

    Klee, G G

    2000-06-01

    Human anti-mouse antibodies (HAMA) are human immunoglobulins with specificity for mouse immunoglobulins. This topic currently is of interest because of the increased use of monoclonal mouse antibodies as diagnostic reagents both for in vitro laboratory measurements and for in vivo imaging studies. Monoclonal mouse antibodies also are being used therapeutically. This short article reviews the production of HAMA in patients receiving monoclonal antibodies and illustrates the potential ways that HAMA can interfere with immunoassay measurements. Methods for measuring and neutralizing HAMA also are discussed.

  10. Utilizing Adventure Activities with Intact Groups: A Sociodramatic Systems Approach to Consultation.

    Science.gov (United States)

    Gillis, H. Lee; Bonney, Warren C.

    1989-01-01

    Describes team-building activities for mental health counselors who function as consultants to staffs, groups, or other intact work systems. Provides rationale for application of strategic systems and sociodrama techniques to an adventure activity that allows a group to metaphorically enact its common issue. Provides example of a consultation…

  11. How-to-Do-It: Cytokinin Induced Cell Division & Differentiation Using Intact Plants.

    Science.gov (United States)

    Bohnsack, Charles W.

    1989-01-01

    Presents a procedure by which cytokinins are used to induce a population of dividing and differentiating cells on the cut surface of the roots of an intact plant. Includes the method used, results, and suggestions for a variety of variables that may be tested. (RT)

  12. The action of cobra venom phospholipase A2 isoenzymes towards intact human erythrocytes

    NARCIS (Netherlands)

    Roelofsen, B.; Sibenius Trip, M.; Verheij, H.M.; Zevenbergen, J.L.

    1980-01-01

    1. 1. Cobra venom phospholipase A2 from three different sources has been fractionated into different isoenzymes by DEAE ion-exchange chromatography. 2. 2. Treatment of intact human erythrocytes with the various isoenzymes revealed significant differences in the degree of phosphatidylcholine hydroly

  13. High incidence of intact or fragmented immunoglobulin in urine of patients with multiple myeloma.

    Science.gov (United States)

    Kraj, Maria; Kruk, Barbara; Lech-Marańda, Ewa; Warzocha, Krzysztof; Prochorec-Sobieszek, Monika

    2015-01-01

    In this prospective study we determined the incidence of intact/fragmented immunoglobulin and Bence Jones protein in urine immunofixation using Sebia reagents and HydrasysTM 2 apparatus and compared the results to concentrations of serum free light chains (FLC) assessed using Siemens BNTM II nephelometer and the immunoassay Freelite (Binding Site) in 289 patients with multiple myeloma at diagnosis. It was found that in one third of IgG, IgA and IgD myeloma patients, intact/fragmented immunoglobulin can be detected in urine and is connected with impaired renal function and reduced survival. Urine immunofixation detects monoclonal protein (FLC and intact/fragmented immunoglobulin) in 66-79% of IgG and IgA myeloma patients while serum FLC immunoassay detect it in 82-94% of IgG and IgA myeloma patients. However, the latter method is inadequate for detection of intact/fragmented immunoglobulin in urine. Serum FLC immunoassay and urine immunofixation are complementary methods in diagnosing and monitoring monoclonal protein in patients with myeloma.

  14. Using Spores for Fusarium spp. Classification by MALDI-Based Intact Cell/Spore Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Wolfgang Winkler

    2012-01-01

    Full Text Available Fusarium is a widespread genus of filamentous fungi and a member of the soil microbial community. Certain subspecies are health threatening because of their mycotoxin production that affects the human and animal food chain. Thus, for early and effective pest control, species identification is of particular interest; however, differentiation on the subspecies level is challenging and time-consuming for this fungus. In the present study, we show the possibilities of intact cell mass spectrometry for spore analysis of 22 different Fusarium strains belonging to six Fusarium subspecies. We found that species differentiation is possible if mass spectrometric analyses are performed under well-defined conditions with fixed parameters. A critical point for analysis is a proper sample preparation of spores, which increases the quality of mass spectra with respect to signal intensity and m/z value variations. It was concluded that data acquistion has to be performed automatically; otherwise, user-specific variations are introduced generating data which cannot fit the existing datasets. Data that show clearly that matrix-assisted laser desorption ionization-based intact cell/intact spore mass spectrometry (IC/ISMS can be applied to differentiate closely related Fusarium spp. are presented. Results show a potential to build a database on Fusarium species for accurate species identification, for fast response in the case of infections in the cornfield. We furthermore demonstrate the high precision of our approach in classification of intact Fusarium species according to the location of their collection.

  15. Collagen fibril size and crimp morphology in ruptured and intact Achilles tendons

    DEFF Research Database (Denmark)

    Magnusson, S P; Qvortrup, K; Larsen, Jytte Overgaard

    2002-01-01

    The present study examined the hypothesis that collagen fibril diameter and crimp angle in ruptured human Achilles tendons differed from that of intact ones. Tissue samples were obtained from the central core (distal core) and the posterior periphery (distal superficial) at the rupture site, and ...... tendon rupture site. Moreover, the lack of symptoms prior to the rupture suggests that clinical tendinopathy is not an etiological factor in complete tendon ruptures.......The present study examined the hypothesis that collagen fibril diameter and crimp angle in ruptured human Achilles tendons differed from that of intact ones. Tissue samples were obtained from the central core (distal core) and the posterior periphery (distal superficial) at the rupture site......, and the proximally intact (proximal superficial) part of the tendon in 10 subjects (38+/-8 years) with a complete tendon rupture. For comparisons corresponding tissue samples were procured from age (38+/-7 years) and gender matched intact Achilles tendons during routine forensic autopsy. The cross-sectional area...

  16. Row-like organization of ATP synthase in intact mitochondria determined by cryo-electron tomography

    NARCIS (Netherlands)

    Dudkina, Natalya V.; Oostergetel, Gert T.; Lewejohann, Dagmar; Braun, Hans-Peter; Boekema, Egbert J.

    2010-01-01

    The fine structure of intact, close-to-spherical mitochondria from the alga Polytomella was visualized by dual-axis cryo-electron tomography. The supramolecular organization of dimeric ATP synthase in the cristae membranes was investigated by averaging subvolumes of tomograms and 3D details at simil

  17. The Father-Child Relationship, Parenting Styles, and Adolescent Risk Behaviors in Intact Families

    Science.gov (United States)

    Bronte-Tinkew, Jacinta; Moore, Kristin A.; Carrano, Jennifer

    2006-01-01

    The father-child relationship and father's parenting style are examined as predictors of first delinquency and substance use, using data from the National Longitudinal Study of Youth 1997, Rounds 1 to 3 (N = 5,345), among adolescents in intact families. Discrete time logistic regressions indicate that a more positive father-child relationship…

  18. Germination of Saccharum ravennae(L.)L.(Poaceae) caryopses and intact spikelets

    Science.gov (United States)

    Ravenna grass, Saccharum ravennae (L.) L., is a robust perennial bunchgrass grown as an ornamental in the United States, but where adapted, has become naturalized in riparian areas. Little is known about the seed size distribution and germination characteristics of caryopses and intact spikelets. ...

  19. Imaging of intact MOF-5 nanocrystals by advanced TEM at liquid

    OpenAIRE

    Wiktor, Christian; Turner, Stuart; Zacher, Denise; Fischer, Roland A.; Tendeloo, van, G.

    2012-01-01

    Abstract: First results on the imaging of intact metalorganic framework (MOF) pores in MOF-5 nanocrystals by aberration corrected transmission electron microscopy (TEM) under liquid nitrogen conditions are presented. The applied technique is certainly transferable to other MOF systems, permitting detailed studies of MOF interfaces, MOFnanoparticle interaction and MOF thin films.

  20. Probing the Intact Cluster Catalysis Concept by Tetrahedral Clusters With Framework Chirality

    Institute of Scientific and Technical Information of China (English)

    G. Süss-Fink; L. Vieille-Petit

    2005-01-01

    @@ 1Results and Discussion In order to bring evidence for or against the hypothesis of catalytic hydrogenation by intact trinuclear arene ruthenium clusters containing an oxo cap, the substrate being hydrogenated inside the hydrophobic pocket spanned by the three arene ligands ("supramolecular cluster catalysis")[1], we synthesized cationic Ru3O clusters (See Fig. 1) with three different arene ligands (intrinsically chiral tetrahedra).

  1. Automated setup for characterization of intact histone tails in Suz12-/- stem cells

    DEFF Research Database (Denmark)

    Sidoli, Simone; Schwämmle, Veit; Hansen, Thomas Aarup

    Epigenetics is defined as the study of heritable changes that occur without modifying the DNA sequence. Histone proteins are crucial components of epigenetic mechanisms and regulation, since they are fundamental for chromatin structure. Mass spectrometry-based proteomics is already an integrated...... developed a high-resolving and automated LC-MS/MS setup to characterize intact histone tails (middle-down strategy)...

  2. Intact School Matching in Education: Exploring the Relative Importance of Focal and Local Matching

    Science.gov (United States)

    Wong, Vivian C.; Hallberg, Kelly; Cook, Thomas D.

    2013-01-01

    The nested data structure inherent in education (i.e. students nested in schools nested in districts) makes intact school matching an appealing approach in observational studies of educational interventions and policies for both theoretical and practical purposes. This paper provides guidance to applied education researchers who are employing…

  3. Family relationships and the psychosocial adjustment of school-aged children in intact families

    NARCIS (Netherlands)

    E.M. Hakvoort; H.M.W. Bos; F. van Balen; J.M.A. Hermanns

    2010-01-01

    The authors investigated whether the quality of three family relationships (i.e., marital, parent-child, sibling) in intact families are associated with each other and with children's psychosocial adjustment. Data were collected by means of maternal and child reports (N = 88) using standardized inst

  4. Urinary felinine excretion in intact male cats is increased by dietary cystine

    NARCIS (Netherlands)

    Hendriks, W.H.; Rutherfurd-Markwick, K.J.; Weidgraaf, K.; Morton, R.H.; Rogers, Q.R.

    2008-01-01

    Felinine is a branched-chain sulfur amino acid present in the urine of certain Felidae, including domestic cats. The objective of the present study was to determine if additional cystine and/or dietary N would increase felinine and N-acetylfelinine excretion by intact male cats fed a low-protein (LP

  5. The action of cobra venom phospholipase A2 isoenzymes towards intact human erythrocytes

    NARCIS (Netherlands)

    Roelofsen, B.; Sibenius Trip, M.; Verheij, H.M.; Zevenbergen, J.L.

    1980-01-01

    1. 1. Cobra venom phospholipase A2 from three different sources has been fractionated into different isoenzymes by DEAE ion-exchange chromatography. 2. 2. Treatment of intact human erythrocytes with the various isoenzymes revealed significant differences in the degree of phosphatidylcholine hydroly

  6. Adolescents' Perception of the Ideal Mate: Its Relationship to Parental Characteristics in Intact and Nonintact Families

    Science.gov (United States)

    Lazar, Amnon; Guttmann, Joseph

    2004-01-01

    The relation of adolescents' experiencing parental divorce, their perception of parents' characteristics, and their perception of the ideal mate's characteristics was investigated. One hundred adolescents from intact families and 79 from nonintact families were asked to rank both the degree to which each of 40 personality traits characterized…

  7. Urinary felinine excretion in intact male cats is increased by dietary cystine

    NARCIS (Netherlands)

    Hendriks, W.H.; Rutherfurd-Markwick, K.J.; Weidgraaf, K.; Morton, R.H.; Rogers, Q.R.

    2008-01-01

    Felinine is a branched-chain sulfur amino acid present in the urine of certain Felidae, including domestic cats. The objective of the present study was to determine if additional cystine and/or dietary N would increase felinine and N-acetylfelinine excretion by intact male cats fed a low-protein

  8. Intactness of cell wall structure controls the in vitro digestion of starch in legumes.

    Science.gov (United States)

    Dhital, Sushil; Bhattarai, Rewati R; Gorham, John; Gidley, Michael J

    2016-03-01

    Increasing the level of starch that is not digested by the end of the small intestine and therefore enters the colon ('resistant starch') is a major opportunity for improving the nutritional profile of foods. One mechanism that has been shown to be successful is entrapment of starch within an intact plant tissue structure. However, the level of tissue intactness required for resistance to amylase digestion has not been defined. In this study, intact cells were isolated from a range of legumes after thermal treatment at 60 °C (starch not gelatinised) or 95 °C (starch gelatinised) followed by hydrolysis using pancreatic alpha amylase. It was found that intact cells, isolated at either temperature, were impervious to amylase. However, application of mechanical force damaged the cell wall and made starch accessible to digestive enzymes. This shows that the access of enzymes to the entrapped swollen starch is the rate limiting step controlling hydrolysis of starch in cooked legumes. The results suggest that a single cell wall could be sufficient to provide an effective delivery of starch to the large intestine with consequent nutritional benefits, provided that mechanical damage during digestion is avoided.

  9. Tympanoplasty in chronic otitis media patients with an intact, but severely retracted malleus: a treatment challenge.

    NARCIS (Netherlands)

    Hol, M.K.S.; Nguyen, D.Q.; Schlegel-Wagner, C.; Pabst, G.; Linder, T.E.

    2010-01-01

    OBJECTIVE: To analyze the outcome of patients with chronic otitis media (COM) with an intact, but markedly medialized ossicular chain, treated by removing the malleus head and interposing an autologous incus and then an underlay myringoplasty. STUDY DESIGN: Retrospective clinical study. SETTING: Ter

  10. Family relationships and the psychosocial adjustment of school-aged children in intact families

    NARCIS (Netherlands)

    Hakvoort, E.M.; Bos, H.M.W.; van Balen, F.; Hermanns, J.M.A.

    2010-01-01

    The authors investigated whether the quality of three family relationships (i.e., marital, parent-child, sibling) in intact families are associated with each other and with children's psychosocial adjustment. Data were collected by means of maternal and child reports (N = 88) using standardized

  11. Compound-specific stable isotope analysis of nitrogen-containing intact polar lipids

    NARCIS (Netherlands)

    Svensson, Elizabeth; Schouten, Stefan; Stam, Axel; Middelburg, Jack J.; Sinninghe Damsté, Jaap S.

    2015-01-01

    Rationale Compound-specific isotope analysis (CSIA) of nitrogen in amino acids has proven a valuable tool in many fields (e.g. ecology). Several intact polar lipids (IPLs) also contain nitrogen, and their nitrogen isotope ratios have the potential to elucidate food-web interactions or metabolic path

  12. Fossilization and degradation of intact polar lipids in deep subsurface sediments: A theoretical approach

    NARCIS (Netherlands)

    Schouten, S.; Middelburg, J.J.; Hopmans, E.C.; Sinninghe Damsté, J.S.

    2010-01-01

    Intact polar membrane lipids (IPLs) are frequently used as markers for living microbial cells in sedimentary environments. The assumption with these studies is that IPLs are rapidly degraded upon cell lysis and therefore IPLs present in sediments are derived from in situ microbial production. We use

  13. Compound-specific stable isotope analysis of nitrogen-containing intact polar lipids

    NARCIS (Netherlands)

    Svensson, E.; Schouten, S.; Stam, A.; Middelburg, J.J.; Sinninghe Damsté, J.S.

    2015-01-01

    RATIONALE: Compound-specific isotope analysis (CSIA) of nitrogen in amino acids has proven a valuable tool in manyfields (e.g. ecology). Several intact polar lipids (IPLs) also contain nitrogen, and their nitrogen isotope ratios have thepotential to elucidate food-web interactions or metabolic pathw

  14. The origin and fate of intact polar lipids in the marine environment

    NARCIS (Netherlands)

    Brandsma, J.

    2011-01-01

    A promising method for the identification, characterization and enumeration of microbial communities in the natural environment is the measurement of intact polar lipids (IPLs), the basic building blocks of biomembranes. These complex molecules are ubiquitous in nature and have several characterist

  15. Designing medical foods for inherited metabolic disorders: why intact protein is superior to amino acids.

    Science.gov (United States)

    Ney, Denise Marie; Etzel, Mark Raymond

    2017-04-01

    Phenylketonuria and tyrosinemia are inherited metabolic disorders characterized by high blood levels of phenylalanine (Phe) or tyrosine (Tyr), due to mutations in genes affecting Phe and Tyr metabolism, respectively. The primary management is a lifelong diet restricted in protein from natural foods in combination with medical foods comprised mixtures of synthetic amino acids. Compliance is often poor after childhood leading to neuropsychological sequela. Glycomacropeptide, an intact 64 amino acid glycophosphopeptide isolated from cheese whey, provides a new paradigm for the management of phenylketonuria and tyrosinemia because glycomacropeptide contains no Phe and Tyr in its pure form, and is also a prebiotic. Medical foods made from glycomacropeptide have been used successfully for the management of phenylketonuria and tyrosinemia. Preclinical and clinical studies demonstrate that intact protein from glycomacropeptide provides a more acceptable and physiologic source of defined protein compared to amino acids in medical foods. For example, harmful gut bacteria were reduced, beneficial short chain fatty acids increased, renal workload decreased, protein utilization increased, and bone fragility decreased using intact protein versus amino acids. Advances in biotechnology will propel the transition from synthetic amino acids to intact proteins for the management of inherited metabolic disorders. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. OXYGEN DEPENDENCE OF PHOTOINHIBITION AT LOW-TEMPERATURE IN INTACT PROTOPLASTS OF VALERIANELLA-LOCUSTA L

    NARCIS (Netherlands)

    VANWIJK, KJ; KRAUSE, GH

    1991-01-01

    Photoinhibition of photosynthesis in vivo is shown to be considerably promoted by O2 under circumstances where energy turnover by photorespiration and photosynthetic carbon metabolism are low. Intact protoplasts of Valerianella locusta L. were photoinhibited by 30 min irradiation with 3000-mu-mol ph

  17. Decubitus grade IV (deep pressure sore) with intact skin in a patient with spinal cord injury

    NARCIS (Netherlands)

    Theunissen, C.C.W.; Zeilstra, J.T.; van Voorst Vader, P.C.; Kardaun, S.H.; Leeman, F.W.J.

    2006-01-01

    Even with intact skin the possibility of pressure sores should not be dismissed. Early recognition of a pressure sore is important for adequate treatment and prevention of progression. Multidisciplinary intervention is essential. A wheelchair patient with spinal cord injury is described, who develop

  18. Non-amphiphilic carbohydrate liquid crystals containing an intact monosaccharide moiety

    NARCIS (Netherlands)

    Smits, E; Engberts, J.B.F.N.; Kellogg, R.M; van Doren, H.A.

    1995-01-01

    A chiral rigid moiety which forms the basis of a new class of non-amphiphilic carbohydrate liquid crystals has been developed. This moiety contains a fully intact glucopyranose ring embedded in a trans-decalin structure. The original carbohydrate is substituted so that only two hydroxyl groups are l

  19. Fertility, pregnancy, and delivery after biventricular repair for pulmonary atresia with an intact ventricular septum.

    NARCIS (Netherlands)

    Drenthen, W.; Pieper, P.G.; Roos-Hesselink, J.W.; Zoon, N.; Voors, A.A.; Mulder, B.J.M.; Dijk, A.P.J. van; Vliegen, H.W.; Sollie, K.M.; Ebels, T.; Veldhuisen, D.J. van

    2006-01-01

    The objective of the present study to investigate fertility, pregnancy, and delivery in women with biventricular repair for pulmonary atresia with an intact ventricular septum (PAIVS). Using a nationwide registry (CONCOR), 37 patients with pulmonary atresia were identified, 6 of whom (aged 21 to 34

  20. Recent advances in the use of non-destructive near infrared spectroscopy on intact olive fruits

    Science.gov (United States)

    The objective of this review is to illustrate the state of the art in the use of non-destructive near infrared (NIR) spectroscopy for quality evaluation of intact fruit in the olive industry. First, the most recent studies regarding the application of non-destructive NIR spectroscopy methods to asse...

  1. Mouse models of medulloblastoma

    Institute of Scientific and Technical Information of China (English)

    Xiaochong Wu; Paul A. Northcott; Sidney Croul; Michael D. Taylor

    2011-01-01

    Medulloblastoma is the most common malignant pediatric brain tumor. Despite its prevalence and importance in pediatric neuro-oncology, the genes and pathways responsible for its initiation, maintenance,and progression remain poorly understood. Genetically engineered mouse models are an essential tool for uncovering the molecular and cellular basis of human diseases, including cancer, and serve a valuable role as preclinical models for testing targeted therapies. In this review, we summarize how such models have been successfully applied to the study of medulloblastoma over the past decade and what we might expect in the coming years.

  2. Simultaneous bicompartmental bucket-handle meniscal tears with intact anterior cruciate ligament: a case report

    Directory of Open Access Journals (Sweden)

    Beris Alexandros E

    2010-02-01

    Full Text Available Abstract Introduction Bucket handle tear of the menisci is a common type of lesion resulting from injury to the knee joint. Bucket handle injury of both menisci in almost all cases is associated with a lesion to either the anterior or the posterior cruciate ligament of the knee joint. We describe a case of acute bucket-handle tear of the medial and lateral menisci with intact anterior and posterior cruciate ligaments in a dancer. To the best of our knowledge, there are no previous reports of this type of injury in the literature. Case presentation A 28-year-old Caucasian Greek woman presented to the emergency department after sustaining an injury to her right knee during dancing. An MRI evaluation demonstrated tears in both menisci of the right knee, while the anterior and posterior cruciate ligaments were found to be intact. A partial medial and lateral meniscectomy was then performed. At a follow-up examination six months after her injury, clinical tests demonstrated that our patient's right knee was stable, had a full range of motion and had no tenderness. She was satisfied with the outcome of the operation and returned to her pre-injury activities. Conclusion We present the first case in the literature that describes a combined bucket-handle injury of both the medial and lateral menisci with an intact anterior cruciate ligament. The clinical examination of the anterior cruciate ligament was unremarkable, with no signs of deficiency or rupture. The posterior cruciate ligament was also intact. On magnetic resonance imaging, the ligaments were visualised as intact in all their length. These findings were confirmed by arthroscopic evaluation.

  3. Dynamic, six-axis stiffness matrix characteristics of the intact intervertebral disc and a disc replacement.

    Science.gov (United States)

    Holsgrove, Timothy P; Gill, Harinderjit S; Miles, Anthony W; Gheduzzi, Sabina

    2015-11-01

    Thorough pre-testing is critical in assessing the likely in vivo performance of spinal devices prior to clinical use. However, there is a lack of data available concerning the dynamic testing of lumbar (porcine model) total disc replacements in all six axes under preload conditions. The aim of this study was to provide new data comparing porcine lumbar spinal specimen stiffness between the intact state and after the implantation of an unconstrained total disc replacement, in 6 degrees of freedom. The dynamic, stiffness matrix testing of six porcine lumbar isolated disc specimens was completed using triangle waves at a test frequency of 0.1 Hz. An axial preload of 500 N was applied during all testing. Specimens were tested both in the intact condition and after the implantation of the total disc replacement. Sixteen key stiffness terms were identified for the comparison of the intact and total disc replacement specimens, comprising the 6 principal stiffness terms and 10 key off-axis stiffness terms. The total disc replacement specimens were significantly different to the intact specimens in 12 of these key terms including all six principal stiffness terms. The implantation of the total disc replacement resulted in a mean reduction in the principal stiffness terms of 100%, 91%, and 98% in lateral bending, flexion-extension, and axial rotation, respectively. The novel findings of this study have demonstrated that the unconstrained, low-friction total disc replacement does not replicate the stiffness of the intact specimens. It is likely that other low-friction total disc replacements would produce similar results due to stiffness being actively minimised as part of the design of low-friction devices, without the introduction of stiffening elements or mechanisms to more accurately replicate the mechanical properties of the natural intervertebral disc. This study has demonstrated, for the first time, a method for the quantitative comparative mechanical function

  4. Aboveground biomass variability across intact and degraded forests in the Brazilian Amazon

    Science.gov (United States)

    Longo, Marcos; Keller, Michael; dos-Santos, Maiza N.; Leitold, Veronika; Pinagé, Ekena R.; Baccini, Alessandro; Saatchi, Sassan; Nogueira, Euler M.; Batistella, Mateus; Morton, Douglas C.

    2016-11-01

    Deforestation rates have declined in the Brazilian Amazon since 2005, yet degradation from logging, fire, and fragmentation has continued in frontier forests. In this study we quantified the aboveground carbon density (ACD) in intact and degraded forests using the largest data set of integrated forest inventory plots (n = 359) and airborne lidar data (18,000 ha) assembled to date for the Brazilian Amazon. We developed statistical models relating inventory ACD estimates to lidar metrics that explained 70% of the variance across forest types. Airborne lidar-ACD estimates for intact forests ranged between 5.0 ± 2.5 and 31.9 ± 10.8 kg C m-2. Degradation carbon losses were large and persistent. Sites that burned multiple times within a decade lost up to 15.0 ± 0.7 kg C m-2 (94%) of ACD. Forests that burned nearly 15 years ago had between 4.1 ± 0.5 and 6.8 ± 0.3 kg C m-2 (22-40%) less ACD than intact forests. Even for low-impact logging disturbances, ACD was between 0.7 ± 0.3 and 4.4 ± 0.4 kg C m-2 (4-21%) lower than unlogged forests. Comparing biomass estimates from airborne lidar to existing biomass maps, we found that regional and pantropical products consistently overestimated ACD in degraded forests, underestimated ACD in intact forests, and showed little sensitivity to fires and logging. Fine-scale heterogeneity in ACD across intact and degraded forests highlights the benefits of airborne lidar for carbon mapping. Differences between airborne lidar and regional biomass maps underscore the need to improve and update biomass estimates for dynamic land use frontiers, to better characterize deforestation and degradation carbon emissions for regional carbon budgets and Reduce Emissions from Deforestation and forest Degradation (REDD+).

  5. First trimester intact hCG as an early marker of trisomy 21: a promise unrecognised?

    Science.gov (United States)

    Spencer, Kevin; Cowans, Nicholas J; Uldbjerg, Niels; Vereecken, Annie; Tørring, Niels

    2008-12-01

    An initial study of trisomy 21 cases showed that prior to 10 weeks, maternal serum levels of intact hCG in the early first trimester are lower than normal. Here we further study the levels prior to and after 10 weeks of gestation to further establish whether or not the intact hCG is effective as a very early screening marker. Fifty-nine samples from pregnancies with trisomy 21 were identified, 31 were collected between the sixth and ninth weeks of gestation and 28 after the tenth week. A series of 629 gestational age-matched samples collected during the same period formed the control group. Intact hCG was measured by a DELFIA assay. The multiples of the median (MoM) in cases (n = 31) collected prior to 10 weeks were 0.79 (CI 0.62-0.98) at a median gestation of 9.1 weeks. Prior to 9 weeks (n = 14) the median was 0.774 (CI 0.54-1.09) at a median gestation of 8.5 weeks. Modelling the detection rate for a 3 or 5% false-positive rate when screening using intact hCG, free beta-hCG and PAPP-A at 8-10 weeks of gestation indicated that 71 or 77% of cases would be detected. More data are needed to establish a secure MoM for intact hCG in pregnancies prior to 10 weeks, before it could be considered a suitable screening marker. Copyright (c) 2008 John Wiley & Sons, Ltd.

  6. Visual speech alters the discrimination and identification of non-intact auditory speech in children with hearing loss.

    Science.gov (United States)

    Jerger, Susan; Damian, Markus F; McAlpine, Rachel P; Abdi, Hervé

    2017-03-01

    Understanding spoken language is an audiovisual event that depends critically on the ability to discriminate and identify phonemes yet we have little evidence about the role of early auditory experience and visual speech on the development of these fundamental perceptual skills. Objectives of this research were to determine 1) how visual speech influences phoneme discrimination and identification; 2) whether visual speech influences these two processes in a like manner, such that discrimination predicts identification; and 3) how the degree of hearing loss affects this relationship. Such evidence is crucial for developing effective intervention strategies to mitigate the effects of hearing loss on language development. Participants were 58 children with early-onset sensorineural hearing loss (CHL, 53% girls, M = 9;4 yrs) and 58 children with normal hearing (CNH, 53% girls, M = 9;4 yrs). Test items were consonant-vowel (CV) syllables and nonwords with intact visual speech coupled to non-intact auditory speech (excised onsets) as, for example, an intact consonant/rhyme in the visual track (Baa or Baz) coupled to non-intact onset/rhyme in the auditory track (/-B/aa or/-B/az). The items started with an easy-to-speechread/B/or difficult-to-speechread/G/onset and were presented in the auditory (static face) vs. audiovisual (dynamic face) modes. We assessed discrimination for intact vs. non-intact different pairs (e.g., Baa:/-B/aa). We predicted that visual speech would cause the non-intact onset to be perceived as intact and would therefore generate more same-as opposed to different-responses in the audiovisual than auditory mode. We assessed identification by repetition of nonwords with non-intact onsets (e.g.,/-B/az). We predicted that visual speech would cause the non-intact onset to be perceived as intact and would therefore generate more Baz-as opposed to az- responses in the audiovisual than auditory mode. Performance in the audiovisual mode showed more same

  7. Burn mouse models

    DEFF Research Database (Denmark)

    Calum, Henrik; Høiby, Niels; Moser, Claus

    2014-01-01

    Severe thermal injury induces immunosuppression, involving all parts of the immune system, especially when large fractions of the total body surface area are affected. An animal model was established to characterize the burn-induced immunosuppression. In our novel mouse model a 6 % third-degree b......Severe thermal injury induces immunosuppression, involving all parts of the immune system, especially when large fractions of the total body surface area are affected. An animal model was established to characterize the burn-induced immunosuppression. In our novel mouse model a 6 % third......-degree burn injury was induced with a hot-air blower. The third-degree burn was confirmed histologically. At 48 h, a decline in the concentration of peripheral blood leucocytes was observed in the group of mice with burn wound. The reduction was ascribed to the decline in concentration of polymorphonuclear...... neutrophil leucocytes and monocytes. When infecting the skin with Pseudomonas aeruginosa, a dissemination of bacteria was observed only in the burn wound group. Histological characterization of the skin showed an increased polymorphonuclear neutrophil granulocytes dominated inflammation in the group of mice...

  8. Cholesterol depletion disorganizes oocyte membrane rafts altering mouse fertilization.

    Directory of Open Access Journals (Sweden)

    Jorgelina Buschiazzo

    Full Text Available Drastic membrane reorganization occurs when mammalian sperm binds to and fuses with the oocyte membrane. Two oocyte protein families are essential for fertilization, tetraspanins and glycosylphosphatidylinositol-anchored proteins. The firsts are associated to tetraspanin-enriched microdomains and the seconds to lipid rafts. Here we report membrane raft involvement in mouse fertilization assessed by cholesterol modulation using methyl-β-cyclodextrin. Cholesterol removal induced: (1 a decrease of the fertilization rate and index; and (2 a delay in the extrusion of the second polar body. Cholesterol repletion recovered the fertilization ability of cholesterol-depleted oocytes, indicating reversibility of these effects. In vivo time-lapse analyses using fluorescent cholesterol permitted to identify the time-point at which the probe is mainly located at the plasma membrane enabling the estimation of the extent of the cholesterol depletion. We confirmed that the mouse oocyte is rich in rafts according to the presence of the raft marker lipid, ganglioside GM1 on the membrane of living oocytes and we identified the coexistence of two types of microdomains, planar rafts and caveolae-like structures, by terms of two differential rafts markers, flotillin-2 and caveolin-1, respectively. Moreover, this is the first report that shows characteristic caveolae-like invaginations in the mouse oocyte identified by electron microscopy. Raft disruption by cholesterol depletion disturbed the subcellular localization of the signal molecule c-Src and the inhibition of Src kinase proteins prevented second polar body extrusion, consistent with a role of Src-related kinases in fertilization via signaling complexes. Our data highlight the functional importance of intact membrane rafts for mouse fertilization and its dependence on cholesterol.

  9. Multi-Axis Prosthetic Knee Resembles Alpine Skiing Movements of an Intact Leg

    Science.gov (United States)

    Demšar, Ivan; Duhovnik, Jože; Lešnik, Blaž; Supej, Matej

    2015-01-01

    The purpose of the study was to analyse the flexion angles of the ski boot, ankle and knee joints of an above-knee prosthesis and to compare them with an intact leg and a control group of skiers. One subject with an above-knee amputation of the right leg and eight healthy subjects simulated the movement of a skiing turn by performing two-leg squats in laboratory conditions. By adding additional loads in proportion to body weight (BW; +1/3 BW, +2/3 BW, +3/3 BW), various skiing regimes were simulated. Change of Flexion Angle (CoFA) and Range of Motion (RoM) in the ski boot, ankle and knee joints were calculated and compared. An average RoM in the skiing boot on the side of prosthesis (4.4 ± 1.1°) was significantly lower compared to an intact leg (5.9 ± 1.8°) and the control group (6.5 ± 2.3°). In the ankle joint, the average RoM was determined to be 13.2±2.9° in the prosthesis, 12.7 ± 2.8° in an intact leg and 14.8±3.6 in the control group. However, the RoM of the knee joint in the prosthesis (42.2 ± 4.2°) was significantly larger than that of the intact leg (34.7 ± 4.4°). The average RoM of the knee joint in the control group was 47.8 ± 5.4°. The influences of additional loads on the kinematics of the lower extremities were different on the side of the prosthesis and on the intact leg. In contrast, additional loads did not produce any significant differences in the control group. Although different CoFAs in the ski boot, ankle and knee joints were used, an above-knee prosthesis with a built-in multi-axis prosthetic knee enables comparable leg kinematics in simulated alpine skiing. Key points The RoM in the ski boot on the side of the prosthetic leg was smaller than the RoM of the intact leg and the control group of healthy subjects. The RoM in the ankle joint of prosthetic leg was comparable to that of the intact leg and the control group of healthy subjects. The RoM in the prosthetic knee joint was greater than the RoM in the knee joint of the

  10. Gankyrin expression during mouse embryogenesis

    Institute of Scientific and Technical Information of China (English)

    秦建民; 刘淑琴; 曾锦章; 李慎菁; 付晓勇; 邱秀华; 吴孟超; 王红阳

    2004-01-01

    Objective: To observe the gene expression of Gankyrin during mouse embryogenesis and reveal the gene biological significance during organs and tissues formation. Methods: The expressions of Gankyrin mRNA in various organs and tissues were detected by in situ hybridization at indicated times during embryogenesis. Results: The expression of Gankyrin mRNA in mouse day 12.5 embryo was mainly in midbrain, interbrain and endbrain; in mouse day 14.5 embryo mainly in midbrain, aorta, liver, gonad, cranium and rib; in mouse day 16.5 embryo mainly in cranium, rib and vertebra;and in mouse day 18.5 embryo mainly in cranium, rib and intestinal mucosa. Conclusion: Gankyrin gene probably participates in the development of the neural tissues (such as midbrain, interbrain and endbrain etc. ), aorta, liver and gonad, intestinal mucosa and bone tissues, which may be closely associated with the function of the organs and tissues.

  11. Evidence does not support absorption of intact solid lipid nanoparticles via oral delivery

    Science.gov (United States)

    Hu, Xiongwei; Fan, Wufa; Yu, Zhou; Lu, Yi; Qi, Jianping; Zhang, Jian; Dong, Xiaochun; Zhao, Weili; Wu, Wei

    2016-03-01

    Whether and to what extent solid lipid nanoparticles (SLNs) can be absorbed integrally via oral delivery should be clarified because it is the basis for elucidation of absorption mechanisms. To address this topic, the in vivo fate of SLNs as well as their interaction with biomembranes is investigated using water-quenching fluorescent probes that can signal structural variations of lipid-based nanocarriers. Live imaging indicates prolonged retention of SLNs in the stomach, whereas in the intestine, SLNs can be digested quickly. No translocation of intact SLNs to other organs or tissues can be observed. The in situ perfusion study shows bioadhesion of both SLNs and simulated mixed micelles (SMMs) to intestinal mucus, but no evidence of penetration of integral nanocarriers. Both SLNs and SMMs exhibit significant cellular uptake, but fail to penetrate cell monolayers. Confocal laser scanning microscopy reveals that nanocarriers mainly concentrate on the surface of the monolayers, and no evidence of penetration of intact vehicles can be obtained. The mucous layer acts as a barrier to the penetration of both SLNs and SMMs. Both bile salt-decoration and SMM formulation help to strengthen the interaction with biomembranes. It is concluded that evidence does not support absorption of intact SLNs via oral delivery.Whether and to what extent solid lipid nanoparticles (SLNs) can be absorbed integrally via oral delivery should be clarified because it is the basis for elucidation of absorption mechanisms. To address this topic, the in vivo fate of SLNs as well as their interaction with biomembranes is investigated using water-quenching fluorescent probes that can signal structural variations of lipid-based nanocarriers. Live imaging indicates prolonged retention of SLNs in the stomach, whereas in the intestine, SLNs can be digested quickly. No translocation of intact SLNs to other organs or tissues can be observed. The in situ perfusion study shows bioadhesion of both SLNs and

  12. Estimation of estradiol in mouse serum samples: evaluation of commercial estradiol immunoassays.

    Science.gov (United States)

    Haisenleder, Daniel J; Schoenfelder, Aleisha H; Marcinko, Elizabeth S; Geddis, Lisa M; Marshall, John C

    2011-11-01

    The University of Virginia Center for Research in Reproduction Ligand Core performed an evaluation of nine commercial estradiol (E2) immunoassays for use with mouse serum. The evaluation had two components. 1) Recovery Studies: a mouse pool was spiked with E2 concentrations across the assay range, and percent recovery and parallelism to the assay standard curve were determined. 2) Correlation Studies: serum pools were collected from intact females, ovariectomized (OVX) and OVX-E2 treated mice and E2 assayed, then measured by gas chromatography/tandem mass spectrometry (GC/MSMS) for comparison to a gold standard method. Recovery results showed that E2 recovery from spiked mouse pools varied greatly (from 640%) among kits tested. However, three kits (DiaSorin Radioimmunoassay, Siemens Double Antibody RIA, and CalBiotech Enzyme Immunoassay) showed reasonable recoveries and parallelism. Data collected from the Correlation Study showed that values from intact, OVX and OVX-E2-treated mouse pools varied by several fold vs. GC/MSMS for most of the kits tested. The DiaSorin RIA and CalBiotech Enzyme Immunoassay Kits showed the best correlation to GC/MSMS. Unfortunately, while this evaluation was ongoing, the DiaSorin Kit was discontinued. In summary, the CalBiotech Kit was the only available assay tested that demonstrated good E2 parallelism to the assay standard curve and accuracy vs. a gold standard method (i.e. GC/MSMS). Also of note, the CalBiotech assay is sensitive and requires minimal sample volume. Therefore, based on these findings the CalBiotech E2 assay has been implemented for use in mouse serum samples within the Ligand Core.

  13. 0.7 and 3 T MRI and sap flow in intact trees: xylem and phloem in action

    NARCIS (Netherlands)

    Homan, N.; Windt, C.W.; Vergeldt, F.J.; Gerkema, E.; As, van H.

    2007-01-01

    Dedicated magnetic resonance imaging (MRI) hardware is described that allows imaging of sap flow in intact trees with a maximal trunk diameter of 4 cm and height of several meters. This setup is used to investigate xylem and phloem flow in an intact tree quantitatively. Due to the fragile gradients

  14. 40 CFR 261.40 - Conditional Exclusion for Used, Intact Cathode Ray Tubes (CRTs) Exported for Recycling.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Conditional Exclusion for Used, Intact Cathode Ray Tubes (CRTs) Exported for Recycling. 261.40 Section 261.40 Protection of Environment...) Exported for Recycling. Used, intact CRTs exported for recycling are not solid wastes if they meet...

  15. 0.7 and 3 T MRI and sap flow in intact trees: xylem and phloem in action

    NARCIS (Netherlands)

    Homan, N.; Windt, C.W.; Vergeldt, F.J.; Gerkema, E.; As, van H.

    2007-01-01

    Dedicated magnetic resonance imaging (MRI) hardware is described that allows imaging of sap flow in intact trees with a maximal trunk diameter of 4 cm and height of several meters. This setup is used to investigate xylem and phloem flow in an intact tree quantitatively. Due to the fragile gradients

  16. Ethanol vapor and saprophytic yeast treatments reduce decay and maintain quality of intact and fresh-cut cherries

    Science.gov (United States)

    The objective of this study was to evaluate the use of an ethanol vapor release pad and a saprophytic yeast (Cryptococcus infirmo-miniatum) to reduce decay and maintain postharvest quality of intact or fresh-cut sweet cherries (Prunus avium L.) cv. Lapins and Bing. Intact or fresh-cut fruit were pac...

  17. Rho kinase inhibitor Y-27632 and Accutase dramatically increase mouse embryonic stem cell derivation.

    Science.gov (United States)

    Zhang, Peng; Wu, Xinglong; Hu, Chunchao; Wang, Pengbo; Li, Xiangyun

    2012-01-01

    Although it has been 30 yr since the development of derivation methods for mouse embryonic stem (ES) cells, the biology of derivation of ES cells is poorly understood and the efficiency varies dramatically between cell lines. Recently, the Rho kinase inhibitor Y-27632 and the cell dissociation reagent Accutase were reported to significantly inhibit apoptosis of human ES cells during passaging. Therefore, in the current study, C57BL/6×129/Sv mouse blastocysts were used to evaluate the effect of the combination of the two reagents instead of using the conventional 129 line in mouse ES cell derivation. The data presented in this study suggests that the combination of Y-27632 and Accutase significantly increases the efficiency of mouse ES cell derivation; furthermore, no negative side effects were observed with Y-27632 and Accutase treatment. The newly established ES cell lines retain stable karyotype, surface markers expression, formed teratomas, and contributed to viable chimeras and germline transmission by tetraploid complementation assay. In addition, Y-27632 improved embryoid body formation of ES cells. During ES cell microinjection, Y-27632 prevented the formation of dissociation-induced cell blebs and facilitates the selection and the capture of intact cells. The methods presented in this study clearly demonstrate that inhibition of Rho kinase with Y-27632 and Accutase dissociation improve the derivation efficiently and reproducibility of mouse ES cell generation which is essential for reducing variability in the results obtained from different cell lines.

  18. Mouse anesthesia and analgesia.

    Science.gov (United States)

    Adams, Sean; Pacharinsak, Cholawat

    2015-03-02

    Providing anesthesia and analgesia for mouse subjects is a common and critical practice in the laboratory setting. These practices are necessary for performing invasive procedures, achieving prolonged immobility for sensitive imaging modalities (magnetic resonance imaging for instance), and providing intra- and post-procedural pain relief. In addition to facilitating the procedures performed by the investigator, the provision of anesthesia and analgesia is crucial for the preservation of animal welfare and for humane treatment of animals used in research. Furthermore, anesthesia and analgesia are important components of animal use protocols reviewed by Institutional Animal Care and Use Committees, requiring careful consideration and planning for the particular animal model. In this article, we provide technical outlines for the investigator covering the provision of anesthesia by two routes (injectable and inhalant), guidelines for monitoring anesthesia, current techniques for recognition of pain, and considerations for administering preventative analgesia. Copyright © 2015 John Wiley & Sons, Inc.

  19. Electrospray ionization Fourier transform mass spectrometric analysis of intact bikunin glycosaminoglycan from normal human plasma.

    Science.gov (United States)

    Laremore, Tatiana N; Leach, Franklin E; Amster, I Jonathan; Linhardt, Robert J

    2011-08-15

    A mixture of glycosaminoglycan (GAG) chains from a plasma proteoglycan bikunin was fractionated using native, continuous-elution polyacrylamide gel electrophoresis, and the resulting fractions were analyzed by electrospray ionization Fourier transform mass spectrometry (ESI FTMS). Molecular mass analysis of the intact GAG afforded information about the length and composition of GAG chains in the mixture. Ambiguity in the interpretation of the intact GAG mass spectra was eliminated by conducting an additional experiment in which the GAG chains of known molecular mass were treated with a GAG-degrading enzyme, chondroitinase ABC, and the digestion products were analyzed by ESI FTMS. The plasma bikunin GAG chains consisted predominantly of odd number of saccharides, although few chains consisting of even number of saccharides were also detected. Majority of the analyzed chains were tetrasulfated or pentasulfated and comprised by 29 to 41 monosaccharides.

  20. Occurrence of iridoid glycosides in in vitro cultures and intact plants of Scrophularia nodosa L.

    Science.gov (United States)

    Sesterhenn, Katja; Distl, Melanie; Wink, Michael

    2007-03-01

    Shoot, root, and callus cultures of Scrophularia nodosa L. (Scrophulariaceae) were established and cultivated in vitro. Iridoid glycosides, such as harpagoside, aucubin, and catalpol were identified by LC-ESI-MS and their contents determined by HPLC. For comparison intact plants of S. nodosa were analysed. In shoot cultures slightly lower amounts of detectable iridoid glycosides (4.36% dry weight) were determined than in the field grown plants (4.88%). Concentration of harpagoside was highest in leaves of field plants (1.05%) and in flowers of in vitro plantlets (1.10%). For aucubin the highest amount was found in the leaves of in vitro plantlets (1.67%) whereas the levels of aucubin in the leaves of field plants were remarkably lower. Catalpol was produced as a trace compound in intact plants and shoot cultures. Callus and root cultures were apparently not able to synthesise iridoid glycosides.

  1. Characterization of the anion sensitive ATPase in intact vacuoles of Kalanchoe diagremontiana

    Energy Technology Data Exchange (ETDEWEB)

    Kobza, J.; Uribe, E.G.

    1986-04-01

    A method for the isolation of intact vacuoles from K. daigremontiana was developed which produced high yields of relatively pure vacuoles as determined by marker enzyme contamination. Upon isolation, the vacuoles were stabilized by the inclusion of 5% (w/v) ficoll. Enzyme activity was insensitive to vanadate and azide but was strongly inhibited by DCCD. Enzyme activity was strictly dependent on the inclusion of Mg/sup 2 +/ and was stimulated by anions as depicted by the series, NO/sub 3//sup -/ < Br/sup -/ < SO/sub 4//sup -/ < HCO/sub 3//sup -/ < Cl/sup -/. It was found that in intact vacuoles the ATPase activity was stimulated by phosphate to a level equivalent to that found with the chloride. The enzyme exhibited Michaelis-Menten kinetics with a Km for Mg-ATP complex of 0.51 mM.

  2. Automated closed-chamber measurements of methane fluxes from intact leaves and trunk of Japanese cypress

    Science.gov (United States)

    Takahashi, Kenshi; Kosugi, Yoshiko; Kanazawa, Akito; Sakabe, Ayaka

    2012-05-01

    Continuous in situ measurements of methane (CH4) fluxes from intact leaves and trunk of Japanese cypress (Chamaecyparis obtusa Sieb. et Zucc) were conducted in a temperate forest from August 2009 to August 2010. An automated closed-chamber system, which was used to evaluate CO2 exchange between the atmosphere and forest ecosystems, was coupled to a laser-based instrument to monitor CH4 concentrations. Temporal changes in CH4 concentrations from the foliage and trunk were measured at one-second intervals during chamber closure to determine CH4 fluxes between the leaf and trunk surfaces and the atmosphere. While recent studies have suggested that some plants emit CH4 under aerobic conditions, emission or uptake of CH4 in detectable amounts with our experimental system, by intact leaves or the trunk of C. obtusa, was not significantly observed throughout the measurement period.

  3. Neurodevelopment. Live imaging of adult neural stem cell behavior in the intact and injured zebrafish brain.

    Science.gov (United States)

    Barbosa, Joana S; Sanchez-Gonzalez, Rosario; Di Giaimo, Rossella; Baumgart, Emily Violette; Theis, Fabian J; Götz, Magdalena; Ninkovic, Jovica

    2015-05-15

    Adult neural stem cells are the source for restoring injured brain tissue. We used repetitive imaging to follow single stem cells in the intact and injured adult zebrafish telencephalon in vivo and found that neurons are generated by both direct conversions of stem cells into postmitotic neurons and via intermediate progenitors amplifying the neuronal output. We observed an imbalance of direct conversion consuming the stem cells and asymmetric and symmetric self-renewing divisions, leading to depletion of stem cells over time. After brain injury, neuronal progenitors are recruited to the injury site. These progenitors are generated by symmetric divisions that deplete the pool of stem cells, a mode of neurogenesis absent in the intact telencephalon. Our analysis revealed changes in the behavior of stem cells underlying generation of additional neurons during regeneration.

  4. 3D Nonlinear Numerical Simulation of Intact and Debonded Reinforced Concrete Beams

    Institute of Scientific and Technical Information of China (English)

    Chen Quan(陈权); Marcus L.

    2004-01-01

    To study the behaviour of reinforced concrete (RC) structures with sections of concrete removed and the reinforcement exposed, 3D nonlinear numerical analysis was performed upon both intact and debonded RC beams by using finite element techniques. The deformational characteristics and the ultimate loads were obtained through numerical models, as well as crack and stress distributions. The failure modes can also be deduced from computational results. Compared with intact beams, the normal assumptions of plane section behaviour is not hold true and the patterns of stress and strain are different in debonded RC beams. The numerical results show good consistency with experimental data. This kind of numerical simulation is a supplement to existing codes.

  5. Percutaneous penetration of uranium in rats after a contamination on intact or wounded skin

    Energy Technology Data Exchange (ETDEWEB)

    Petitot, F.; Gautier, C.; Moreels, A.M.; Frelon, S.; Paquet, F. [Institut de Radioprotection et de Surete Nucleaire, Laboratoire de Radiotoxicologie Experimentale, IRSN/DRPH/SRBE/LRTOX, Site du Tricastin, B.P. 166, 26702 Pierrelatte Cedex (France)

    2007-07-01

    The aim of this work is to assess in vivo in a hairless rat model, the percutaneous diffusion of uranium through intact or wounded rat skin. Six types of wounds were simulated by excoriation and burns with 10 N HF, 2, 5 and 14 N HNO{sub 3} and 10 N NaOH on anesthetized hairless rats. Percutaneous penetration through wounded skin towards blood and subsequent urinary excretion of uranium was followed in vivo during 24 h. The influence of the physicochemical form (solution or powder) of uranyl nitrate (UN) on its percutaneous diffusion was also investigated. UN, even as a powder, can diffuse through intact skin. The presence of uranium in blood is more persistent and its urinary elimination is slower after an HF burn than after an HNO{sub 3} burn. Excoriation increases dramatically percutaneous absorption of UN. Thus, percutaneous diffusion of UN is largely dependent on skin barrier integrity with a particular importance of stratum corneum. (authors)

  6. Slow passive diffusion of NAD+ between intact isolated plant mitochondria and suspending medium.

    Science.gov (United States)

    Neuburger, M; Douce, R

    1983-11-15

    Isolated potato (Solanum tuberosum) tuber mitochondria purified by isopycnic centrifugation in density gradients of Percoll were found to be highly intact, to be devoid of extramitochondrial contaminations and to retain a high rate of O2 consumption. When suspended in a medium that avoided rupture of the outer membrane, intact purified mitochondria progressively lost their NAD+ content by passive diffusion. This led to a slow decrease of oxoglutarate-dependent O2 consumption by isolated mitochondria. Addition of NAD+ to the medium restored the initial State-3 rate of oxoglutarate oxidation. The rate of NAD+ accumulation in the matrix space was concentration-dependent, exhibited Michaelis-Menten kinetics and was strongly inhibited by the analogue N-4-azido-2-nitrophenyl-4-aminobutyryl-NAD+.

  7. Persistent Truncus Arteriosus With Intact Ventricular Septum: Clinical, Hemodynamic and Short-term Surgical Outcome

    Directory of Open Access Journals (Sweden)

    Gholamhossein Ajami

    2015-10-01

    Full Text Available Introduction: Truncus arteriosus with intact ventricular septum is a rare and unique variant of persistent truncus arteriosus (PTA which usually presents with central cyanosis and congestive heart failure in neonate and early infancy. Associated cardiac and non-cardiac anomalies may affect morbidity and mortality of these patients. Case Presentation: We describe clinical presentation, echocardiography and angiographic features of a 7-month old boy with PTA and intact ventricular septum who underwent surgical repair of the anomaly at our institution. Operative findings, surgical procedure and short-term outcome are reported. Conclusions: While our patient had systemic pulmonary arterial pressure at the time of complete surgical repair, it was improved after surgery.

  8. Intact-Brain Analyses Reveal Distinct Information Carried by SNc Dopamine Subcircuits.

    Science.gov (United States)

    Lerner, Talia N; Shilyansky, Carrie; Davidson, Thomas J; Evans, Kathryn E; Beier, Kevin T; Zalocusky, Kelly A; Crow, Ailey K; Malenka, Robert C; Luo, Liqun; Tomer, Raju; Deisseroth, Karl

    2015-07-30

    Recent progress in understanding the diversity of midbrain dopamine neurons has highlighted the importance--and the challenges--of defining mammalian neuronal cell types. Although neurons may be best categorized using inclusive criteria spanning biophysical properties, wiring of inputs, wiring of outputs, and activity during behavior, linking all of these measurements to cell types within the intact brains of living mammals has been difficult. Here, using an array of intact-brain circuit interrogation tools, including CLARITY, COLM, optogenetics, viral tracing, and fiber photometry, we explore the diversity of dopamine neurons within the substantia nigra pars compacta (SNc). We identify two parallel nigrostriatal dopamine neuron subpopulations differing in biophysical properties, input wiring, output wiring to dorsomedial striatum (DMS) versus dorsolateral striatum (DLS), and natural activity patterns during free behavior. Our results reveal independently operating nigrostriatal information streams, with implications for understanding the logic of dopaminergic feedback circuits and the diversity of mammalian neuronal cell types.

  9. Forest Loss in Protected Areas and Intact Forest Landscapes: A Global Analysis.

    Directory of Open Access Journals (Sweden)

    Matias Heino

    Full Text Available In spite of the high importance of forests, global forest loss has remained alarmingly high during the last decades. Forest loss at a global scale has been unveiled with increasingly finer spatial resolution, but the forest extent and loss in protected areas (PAs and in large intact forest landscapes (IFLs have not so far been systematically assessed. Moreover, the impact of protection on preserving the IFLs is not well understood. In this study we conducted a consistent assessment of the global forest loss in PAs and IFLs over the period 2000-2012. We used recently published global remote sensing based spatial forest cover change data, being a uniform and consistent dataset over space and time, together with global datasets on PAs' and IFLs' locations. Our analyses revealed that on a global scale 3% of the protected forest, 2.5% of the intact forest, and 1.5% of the protected intact forest were lost during the study period. These forest loss rates are relatively high compared to global total forest loss of 5% for the same time period. The variation in forest losses and in protection effect was large among geographical regions and countries. In some regions the loss in protected forests exceeded 5% (e.g. in Australia and Oceania, and North America and the relative forest loss was higher inside protected areas than outside those areas (e.g. in Mongolia and parts of Africa, Central Asia, and Europe. At the same time, protection was found to prevent forest loss in several countries (e.g. in South America and Southeast Asia. Globally, high area-weighted forest loss rates of protected and intact forests were associated with high gross domestic product and in the case of protected forests also with high proportions of agricultural land. Our findings reinforce the need for improved understanding of the reasons for the high forest losses in PAs and IFLs and strategies to prevent further losses.

  10. Forest Loss in Protected Areas and Intact Forest Landscapes: A Global Analysis.

    Science.gov (United States)

    Heino, Matias; Kummu, Matti; Makkonen, Marika; Mulligan, Mark; Verburg, Peter H; Jalava, Mika; Räsänen, Timo A

    2015-01-01

    In spite of the high importance of forests, global forest loss has remained alarmingly high during the last decades. Forest loss at a global scale has been unveiled with increasingly finer spatial resolution, but the forest extent and loss in protected areas (PAs) and in large intact forest landscapes (IFLs) have not so far been systematically assessed. Moreover, the impact of protection on preserving the IFLs is not well understood. In this study we conducted a consistent assessment of the global forest loss in PAs and IFLs over the period 2000-2012. We used recently published global remote sensing based spatial forest cover change data, being a uniform and consistent dataset over space and time, together with global datasets on PAs' and IFLs' locations. Our analyses revealed that on a global scale 3% of the protected forest, 2.5% of the intact forest, and 1.5% of the protected intact forest were lost during the study period. These forest loss rates are relatively high compared to global total forest loss of 5% for the same time period. The variation in forest losses and in protection effect was large among geographical regions and countries. In some regions the loss in protected forests exceeded 5% (e.g. in Australia and Oceania, and North America) and the relative forest loss was higher inside protected areas than outside those areas (e.g. in Mongolia and parts of Africa, Central Asia, and Europe). At the same time, protection was found to prevent forest loss in several countries (e.g. in South America and Southeast Asia). Globally, high area-weighted forest loss rates of protected and intact forests were associated with high gross domestic product and in the case of protected forests also with high proportions of agricultural land. Our findings reinforce the need for improved understanding of the reasons for the high forest losses in PAs and IFLs and strategies to prevent further losses.

  11. Ion Channel Photoswitch Reveals Crosstalk between Intact and Injured Nociceptive Neurons after Nerve Injury

    OpenAIRE

    Herold, Christian

    2015-01-01

    The development of novel techniques utilizing the advantages of light has created an optical revolution for neuroscience research. Controlling and probing neuronal function with light has provided unprecedented insights by being able to manipulate many neurons simultaneously in intact circuits and living organisms.In my dissertation research, I used novel optical methods to probe the cellular permeability of sensory neuron populations. Primary nociceptive afferents detect, modulate and integr...

  12. Cobalt Oxide Nanoparticles: Behavior towards Intact and Impaired Human Skin and Keratinocytes Toxicity

    OpenAIRE

    Marcella Mauro; Matteo Crosera; Marco Pelin; Chiara Florio; Francesca Bellomo; Gianpiero Adami; Piero Apostoli; Giuseppe Palma; Massimo Bovenzi; Marco Campanini; Francesca Larese Filon

    2015-01-01

    Skin absorption and toxicity on keratinocytes of cobalt oxide nanoparticles (Co3O4NPs) have been investigated. Co3O4NPs are commonly used in industrial products and biomedicine. There is evidence that these nanoparticles can cause membrane damage and genotoxicity in vitro, but no data are available on their skin absorption and cytotoxicity on keratinocytes. Two independent 24 h in vitro experiments were performed using Franz diffusion cells, using intact (experiment 1) and needle-abraded huma...

  13. In vitro absorption of metal powders through intact and damaged human skin.

    Science.gov (United States)

    Filon, Francesca Larese; D'Agostin, Flavia; Crosera, Matteo; Adami, Gianpiero; Bovenzi, Massimo; Maina, Giovanni

    2009-06-01

    The bioavailability of metals, which are known as important contact allergens, is decisive for the development and the maintenance of contact dermatitis. The aim of this study was to evaluate the percutaneous penetration of metal powders of cobalt (Co), nickel (Ni) and chromium (Cr) and the effect of skin lesions on skin absorption. In vitro permeation experiments were performed using the Franz diffusion cells with intact and damaged human skin. Physiological solution was used as receiving phase and metal powders (Co, Ni and Cr) dispersed in synthetic sweat at pH 4.5 were applied as donor phase to the outer surface of the skin for 24h. The amount of each metal permeating the skin was analysed by electro-thermal atomic absorption spectroscopy (ETAAS). Donor solution analysis demonstrated that metals were present as ions. Measurements of metals skin content were also exploited. Median Co and Ni concentrations found in the receiving phase were significantly higher when Co and Ni powders were applied on the abraded skin than after application on the intact skin (3566 and 2631ngcm(-2) vs. 8.4 and 31ngcm(-2), respectively). No significant difference was found in Cr permeation through intact and damaged skin. The measurement of metals skin content showed that Co, Ni and Cr concentrations were significantly higher in the damaged skin than in the intact skin. Co and Ni ions concentrations increased significantly when the donor solutions were applied on the damaged skin, while Cr ions concentrations did not increase. This study demonstrated that Co and Ni powders can permeate through damaged skin more easily than Cr powder, which has probably a stronger skin proteins binding capacity. Therefore, our results suggest that is necessary to prevent skin contamination when using toxic substances because a small injury to the skin barrier can significantly increase skin absorption.

  14. DETERMINATION OF THE MINIMUM INTACT DIMENSIONS AVAILABLE IN PRACTICAL APPLICATIONS OF LASER CUTTING

    Directory of Open Access Journals (Sweden)

    Peter Gyorgy HORVATH

    2016-06-01

    Full Text Available The reuse of wastes is one of the main requirements of sustainable wood processing. Technologies available for waste management however are currently suited to the reuse of large quantities. This study investigates the utilisation of individual pieces of sawn wood and veneer wastes by laser cutting, with special emphasis on the search for the practical minimum intact dimension that can be produced by laser cutting

  15. Expression of the intact C4 type pepc gene cloned from maize in transgenic winter wheat

    Institute of Scientific and Technical Information of China (English)

    CHEN Xuqing; ZHANG Xiaodong; LIANG Rongqi; ZHANG Liquan; YANG Fengping; CAO Mingqing

    2004-01-01

    Maize intact C4-pepc gene was amplified through LA-PCR and successfully sub-cloned into modified vector pGreen0029 to form a stable expression construct named as Pbac214 (12 kb), which contains CaMV 35S promoter driven bar gene as selection marker. Comparing the cloned DNA sequences (6.7 kb) with published maize C4-pepc gene (GenBank accession E17154) sequences, the identity of DNA sequence alignment is 98.96%. There are only 49 differences between these two intact DNA sequences, of which 13 occur in the region of promoter, 18 in introns, and 18 in exons. The homology of Mrna sequence alignment is 99.38%, and the putative amino acids sequence identity is 99.38%. There are only 15 differences between these two Mrna, and these differences bring 4 sites mutant on the putative amino acids of PEPC protein. Through biolistic bombardment of PDS1000/He system, expression vector Pbac214 has been transformed into winter wheat. Southern blotting results show that the intact C4-pepc gene has been integrated into genome of winter wheat. SDS-PAGE analysis of leaf soluble protein in transgenic wheat showed that the intact C4-pepc gene was well transcribed, spliced and translated as in maize. The enzyme activity of leaf PEPC in transgenic wheat has been detected. The activities of leaf PEPC increased over 3-5 times in some transgenic plants. The data of photosynthesis rate and transpiration rate of transgenic wheat flag leaves showed that the C4-pepc gene can increase the photosynthesis rate and transpiration rate of transgenic wheat.

  16. Large Right Ventricular Clot in Pulmonary Atresia With Intact Ventricular Septum: In Defense of Biventricular Approach.

    Science.gov (United States)

    Dutta, Nilanjan; Ghosh, Rajarshi; Awasthy, Neeraj; Iyer, Parvathi U; Girotra, Sumir; Iyer, Krishna S

    2016-09-01

    Thrombus formation within the right ventricle (RV) in the setting of pulmonary atresia with intact ventricular septum (PAIVS) is not a very common occurrence and can be catastrophic. We present the case of a seven-month-old child with PAIVS and RV clot who successfully underwent biventricular repair. We discuss the interesting case and the rationale for management by means of biventricular repair over single ventricle repair when feasible in such a setting.

  17. Atomic force microscopy of swelling and hardening of intact erythrocytes fixed on substrate

    Science.gov (United States)

    Khalisov, M. M.; Timoshchuk, K. I.; Ankudinov, A. V.; Timoshenko, T. E.

    2017-02-01

    Peak force measurements with the aid of atomic force microscopy are used to quantitatively map nanomechanical properties of intact erythrocytes of rats under conditions that are close to physiological conditions. Erythrocytes that are immobilized on the substrate preliminary processed using poly-L-lysine predominantly exhibit plane shape. However, cells may also exhibit stepwise transformation to semispherical objects with an increase in volume and hardening. Possible reasons for such transformations are discussed.

  18. Efficient and simple electro-transformation of intact cells for the basidiomycetous fungus Pseudozyma hubeiensis

    OpenAIRE

    Konishi, Masaaki; Yoshida, Yuta; Ikarashi, Mizuki; Horiuchi, Junichi; 小西, 正朗

    2015-01-01

    Objective: An electroporation procedure for the species was investigated to develop an efficient transformation method for the basidiomycetous fungus Pseudozyma hubeiensis SY62, a strong biosurfactant-producing host. Results: A plasmid, pUXV1emgfp including green fluorescence protein as a reporter gene, was constructed to determine the transformation and expression of foreign genes. Optimal electroporation conditions achieved 44.8 transformants μg−1 plasmid competency (intact cells) without p...

  19. Regulation of Ribulose Bisphosphate Carboxylase Activity in Intact Wheat Leaves by Light, CO2, and Temperature

    OpenAIRE

    MÄCHLER, F.; NÖSBERGER, J.

    2017-01-01

    The activity of the enzyme ribulose bisphosphate carboxylase (RuBPCase) was estimated after rapidly extracting it from intact wheat leaves pretreated under different light and CO2 levels. No HCO3− was added to the extraction buffer since it is shown to inhibit RuBPCase. The activity increased as light intensity or CO2 concentration during pretreatment was increased. Enzyme activity increased as temperature during pretreatment was decreased. Light activation did not affect the affinity of RuBP...

  20. Investigation of Ion Transmission Effects on Intact Protein Quantification in a Triple Quadrupole Mass Spectrometer

    Science.gov (United States)

    Wang, Evelyn H.; Appulage, Dananjaya Kalu; McAllister, Erin A.; Schug, Kevin A.

    2017-09-01

    Recently, direct intact protein quantitation using triple quadrupole mass spectrometry (QqQ-MS) and multiple reaction monitoring (MRM) was demonstrated (J. Am. Soc. Mass Spectrom. 27, 886-896 (2016)). Even though QqQ-MS is known to provide extraordinary detection sensitivity for quantitative analysis, we found that intact proteins exhibited a less than 5% ion transmission from the first quadrupole to the third quadrupole mass analyzer in the presence of zero collision energy (ZCE). With the goal to enhance intact protein quantitation sensitivity, ion scattering effects, proton transfer effects, and mass filter resolution widths were examined for their contributions to the lost signal. Protein standards myoglobin and ubiquitin along with small molecules reserpine and vancomycin were analyzed together with various collision induced dissociation (CID) gases (N2, He, and Ar) at different gas pressures. Mass resolution settings played a significant role in reducing ion transmission signal. By narrowing the mass resolution window by 0.35 m/z on each side, roughly 75%-90% of the ion signal was lost. The multiply charged proteins experienced additional proton transfer effects, corresponding to 10-fold signal reduction. A study of increased sensitivity of the method was also conducted with various MRM summation techniques. Although the degree of enhancement was analyte-dependent, an up to 17-fold increase in sensitivity was observed for ubiquitin using a summation of multiple MRM transitions. Biological matrix, human urine, and equine plasma were spiked with proteins to demonstrate the specificity of the method. This study provides additional insight into optimizing the use and sensitivity of QqQ-MS for intact protein quantification. [Figure not available: see fulltext.

  1. Structural Studies on Intact Clostridium botulinum Neurotoxins Complexed with Inhibitors Leading to Drug Design

    Science.gov (United States)

    2008-02-01

    Clostridium botulinum Neurotoxins Complexed with Inhibitors Leading to Drug Design PRINCIPAL INVESTIGATOR: Subramanyam Swaminathan...Inhibitors Leading to Drug Design 5b. GRANT NUMBER DAMD17-02-2-0011 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) Subramanyam Swaminathan, Ph.D. 5d...on Intact Clostridium botulinum Neurotoxins Complexed with Inhibitors Leading to Drug Design Annual Report for the Period ending January 2008

  2. Cyanide reduction by nitrogenase in intact cells of Rhodopseudomonas gelatinose Molisch.

    Science.gov (United States)

    Materassi, R; Balloni, W; Florenzano, G

    1977-01-01

    Intact cells of Rhodopseudomonas gelatinosa, grown in N-free medium, reduce cyanide to methane and ammonia at a rate 5--8 times lower than acetylene. Cyanide reduction in whole cells is distinctly more sensitive to cyanide inhibition compared with enzyme preparations. These results are discussed in view of the exploitation of nitrogen-fixing photobacteria in the anaerobic detoxification of cyanide-containing wastewaters.

  3. The Effect of Self-Transcendence on Depression in Cognitively Intact Nursing Home Patients

    OpenAIRE

    Gørill Haugan; Siw Tone Innstrand

    2012-01-01

    Aims. This study's aim was to test the effects of self-transcendence on depression among cognitively intact nursing home patients. Background. Depression is considered the most frequent mental disorder among the elderly population. Specifically, the depression rate among nursing home patients is three to four times higher than that among community-dwelling elderly. Therefore, finding new and alternative ways to prevent and decrease depression is of great importance for nursing home patients' ...

  4. Bioengineered human IAS reconstructs with functional and molecular properties similar to intact IAS

    OpenAIRE

    Singh, Jagmohan; Rattan, Satish

    2012-01-01

    Because of its critical importance in rectoanal incontinence, we determined the feasibility to reconstruct internal anal sphincter (IAS) from human IAS smooth muscle cells (SMCs) with functional and molecular attributes similar to the intact sphincter. The reconstructs were developed using SMCs from the circular smooth muscle layer of the human IAS, grown in smooth muscle differentiation media under sterile conditions in Sylgard-coated tissue culture plates with central Sylgard posts. The bas...

  5. Fourier transform infrared spectroscopic study of intact cells of the nitrogen-fixing bacterium Azospirillum brasilense

    Science.gov (United States)

    Kamnev, A. A.; Ristić, M.; Antonyuk, L. P.; Chernyshev, A. V.; Ignatov, V. V.

    1997-06-01

    The data of Fourier transform infrared (FTIR) spectroscopic measurements performed on intact cells of the soil nitrogen-fixing bacterium Azospirillum brasilense grown in a standard medium and under the conditions of an increased metal uptake are compared and discussed. The structural FTIR information obtained is considered together with atomic absorption spectrometry (AAS) data on the content of metal cations in the bacterial cells. Some methodological aspects concerning preparation of bacterial cell samples for FTIR measurements are also discussed.

  6. MouseCyc: a curated biochemical pathways database for the laboratory mouse

    OpenAIRE

    Evsikov, Alexei V.; Dolan, Mary E.; Genrich, Michael P; Patek, Emily; Bult, Carol J.

    2009-01-01

    Linking biochemical genetic data to the reference genome for the laboratory mouse is important for comparative physiology and for developing mouse models of human biology and disease. We describe here a new database of curated metabolic pathways for the laboratory mouse called MouseCyc . MouseCyc has been integrated with genetic and genomic data for the laboratory mouse available from the Mouse Genome Informatics database and with pathway data from other organisms, including human.

  7. Whole mouse cryo-imaging

    Science.gov (United States)

    Wilson, David; Roy, Debashish; Steyer, Grant; Gargesha, Madhusudhana; Stone, Meredith; McKinley, Eliot

    2008-03-01

    The Case cryo-imaging system is a section and image system which allows one to acquire micron-scale, information rich, whole mouse color bright field and molecular fluorescence images of an entire mouse. Cryo-imaging is used in a variety of applications, including mouse and embryo anatomical phenotyping, drug delivery, imaging agents, metastastic cancer, stem cells, and very high resolution vascular imaging, among many. Cryo-imaging fills the gap between whole animal in vivo imaging and histology, allowing one to image a mouse along the continuum from the mouse -> organ -> tissue structure -> cell -> sub-cellular domains. In this overview, we describe the technology and a variety of exciting applications. Enhancements to the system now enable tiled acquisition of high resolution images to cover an entire mouse. High resolution fluorescence imaging, aided by a novel subtraction processing algorithm to remove sub-surface fluorescence, makes it possible to detect fluorescently-labeled single cells. Multi-modality experiments in Magnetic Resonance Imaging and Cryo-imaging of a whole mouse demonstrate superior resolution of cryo-images and efficiency of registration techniques. The 3D results demonstrate the novel true-color volume visualization tools we have developed and the inherent advantage of cryo-imaging in providing unlimited depth of field and spatial resolution. The recent results continue to demonstrate the value cryo-imaging provides in the field of small animal imaging research.

  8. Predictors of attachment security in preschool children from intact and divorced families.

    Science.gov (United States)

    Nair, Hira; Murray, Ann D

    2005-09-01

    The authors selected 58 mother-child dyads from divorced and intact families to participate in a study on the impact of divorce on preschoolers' attachment security. The authors explored pathways that lead to security of attachment. They found that mothers from divorced families were younger, had lower income levels, and had lower levels of education compared with their intact counterparts. Divorced mothers also reported significantly higher levels of stress, depression, need for social support, and conflict with their spouses. Mothers from intact families were more likely to use positive (authoritative) parenting styles compared with divorced mothers. Children in the divorced group had lower security scores on the Attachment Q-Set instrument (E. Waters, 1995). Regression analyses indicated that parenting style made a direct (independent) contribution to attachment security. In addition, temperament was related to attachment security, but temperament did not diminish the association of parenting style with attachment security. Furthermore, regression analyses indicated that the relationship of divorce to attachment security was mediated by parenting style.

  9. Biomechanical Comparison Of Intact Lumbar Lamp Spine And Endoscopic Discectomized Lamp Spine

    Directory of Open Access Journals (Sweden)

    Ahmet Karakasli

    2012-06-01

    Full Text Available Objectives: Purpose of current study was biomechanical comparison of changes wrought on motion segments after minimally invasive percutan endoscopic discectomized and intact spine. Materials and Methods: We prepared ten fresh-frozen lamb spines were used for this study. The spine of each specimen was dissected between L4-L5. The biomechanical tests for both intact spine and discectomized spine were performed by using axial compression testing machine (AG-I 10 kN, Shimadzu, Japanese. The axial compression was applied to all specimens with the loading speed of 5 mm/min. 8400 N/mm moment was applied to each specimen to achieve flexion and extension motions, right and left bending by a specially designed fixture. Results: In axial compression and flexion tests, the specimens were more stable according to displacement values. The displacement values of sectioned specimens were closer to intact specimens. Only displacement values of left-bending anteroposterior test for both situations were significant (0.05 ;#8805; P. Conclusion: PTED hasn't biomechanical and cilinical disadvantages. Endoscopic discectomy hadn't any disadvantages in stability. Only anterior-posterior displacement values of left bending test were statistically significant. We consider that cause of these results were due to the fact that all specimens had percutan transforaminal endoscopic discectomy (PTED from left side.

  10. Membrane-bound ATPase of intact vacuoles and tonoplasts isolated from mature plant tissue

    Energy Technology Data Exchange (ETDEWEB)

    Lin, W.; Wagner, G.J.; Siegelman, H.W.; Hind, G.

    1977-01-01

    Intact vacuoles were isolated from petals of Hippeastrum and Tulipa (Wagner, G. J. and Siegelman, H. W. (1975) Science 190, 1298 to 1299). The ATPase activity of fresh vacuole suspensions was found to be 2 to 3 times that of protoplasts from the same tissue. 70 to 80% of the ATPase activity of intact vacuoles was recovered in tonoplast preparations. The antibiotic Dio-9 at 6 ..mu..g/10/sup 6/ vacuoles or protoplasts causes 40% inhibition. However, only the protoplast ATPase is sensitive to oligomycin. N,N'-dicyclohexylcarbondiimide (DCCD) slightly stimulates ATPase activity in both vacuole and protoplast suspensions, whereas ethyl-3-(3-dimethylaminopropyl carbodiimide) (EDAC) strongly inhibits. Spectrophotometric studies show that in the petal the vacuolar contents have a pH of 4.0 for Tulipa and 4.3 for Hippeastrum, whereas the intact isolated vacuole has an internal pH of 7.0 (in pH 8.0 buffer) for Tulipa and about 7.3 for Hippeastrum. Internal ion concentrations of 150, 46, 30, 30 and 6 mM were found for K/sup +/, Na/sup +/, Mg/sup 2 +/, Cl/sup -/, and Ca/sup 2 +/ respectively, which are about the same as those in protoplasts.

  11. Multiple Reaction Monitoring for Direct Quantitation of Intact Proteins Using a Triple Quadrupole Mass Spectrometer.

    Science.gov (United States)

    Wang, Evelyn H; Combe, Peter C; Schug, Kevin A

    2016-05-01

    Methods that can efficiently and effectively quantify proteins are needed to support increasing demand in many bioanalytical fields. Triple quadrupole mass spectrometry (QQQ-MS) is sensitive and specific, and it is routinely used to quantify small molecules. However, low resolution fragmentation-dependent MS detection can pose inherent difficulties for intact proteins. In this research, we investigated variables that affect protein and fragment ion signals to enable protein quantitation using QQQ-MS. Collision induced dissociation gas pressure and collision energy were found to be the most crucial variables for optimization. Multiple reaction monitoring (MRM) transitions for seven standard proteins, including lysozyme, ubiquitin, cytochrome c from both equine and bovine, lactalbumin, myoglobin, and prostate-specific antigen (PSA) were determined. Assuming the eventual goal of applying such methodology is to analyze protein in biological fluids, a liquid chromatography method was developed. Calibration curves of six standard proteins (excluding PSA) were obtained to show the feasibility of intact protein quantification using QQQ-MS. Linearity (2-3 orders), limits of detection (0.5-50 μg/mL), accuracy (protein. Sensitivities for different proteins varied considerably. Biological fluids, including human urine, equine plasma, and bovine plasma were used to demonstrate the specificity of the approach. The purpose of this model study was to identify, study, and demonstrate the advantages and challenges for QQQ-MS-based intact protein quantitation, a largely underutilized approach to date.

  12. Multiple Reaction Monitoring for Direct Quantitation of Intact Proteins Using a Triple Quadrupole Mass Spectrometer

    Science.gov (United States)

    Wang, Evelyn H.; Combe, Peter C.; Schug, Kevin A.

    2016-05-01

    Methods that can efficiently and effectively quantify proteins are needed to support increasing demand in many bioanalytical fields. Triple quadrupole mass spectrometry (QQQ-MS) is sensitive and specific, and it is routinely used to quantify small molecules. However, low resolution fragmentation-dependent MS detection can pose inherent difficulties for intact proteins. In this research, we investigated variables that affect protein and fragment ion signals to enable protein quantitation using QQQ-MS. Collision induced dissociation gas pressure and collision energy were found to be the most crucial variables for optimization. Multiple reaction monitoring (MRM) transitions for seven standard proteins, including lysozyme, ubiquitin, cytochrome c from both equine and bovine, lactalbumin, myoglobin, and prostate-specific antigen (PSA) were determined. Assuming the eventual goal of applying such methodology is to analyze protein in biological fluids, a liquid chromatography method was developed. Calibration curves of six standard proteins (excluding PSA) were obtained to show the feasibility of intact protein quantification using QQQ-MS. Linearity (2-3 orders), limits of detection (0.5-50 μg/mL), accuracy (protein. Sensitivities for different proteins varied considerably. Biological fluids, including human urine, equine plasma, and bovine plasma were used to demonstrate the specificity of the approach. The purpose of this model study was to identify, study, and demonstrate the advantages and challenges for QQQ-MS-based intact protein quantitation, a largely underutilized approach to date.

  13. Rapid Prediction of Moisture Content in Intact Green Coffee Beans Using Near Infrared Spectroscopy

    Directory of Open Access Journals (Sweden)

    Adnan Adnan

    2017-05-01

    Full Text Available Moisture content (MC is one of the most important quality parameters of green coffee beans. Therefore, its fast and reliable measurement is necessary. This study evaluated the feasibility of near infrared (NIR spectroscopy and chemometrics for rapid and non-destructive prediction of MC in intact green coffee beans of both Coffea arabica (Arabica and Coffea canephora (Robusta species. Diffuse reflectance (log 1/R spectra of intact beans were acquired using a bench top Fourier transform NIR instrument. MC was determined gravimetrically according to The International Organization for Standardization (ISO 6673. Samples were split into subsets for calibration (n = 64 and independent validation (n = 44. A three-component partial least squares regression (PLSR model using raw NIR spectra yielded a root mean square error of prediction (RMSEP of 0.80% MC; a four component PLSR model using scatter corrected spectra yielded a RMSEP of 0.57% MC. A simplified PLS model using seven selected wavelengths (1155, 1212, 1340, 1409, 1724, 1908, and 2249 nm yielded a similar accuracy (RMSEP: 0.77% MC which opens the possibility of creating cheaper NIR instruments. In conclusion, NIR diffuse reflectance spectroscopy appears to be suitable for rapid and reliable MC prediction in intact green coffee; no separate model for Arabica and Robusta species is needed.

  14. Rapid Prediction of Moisture Content in Intact Green Coffee Beans Using Near Infrared Spectroscopy.

    Science.gov (United States)

    Adnan, Adnan; Hörsten, Dieter von; Pawelzik, Elke; Mörlein, And Daniel

    2017-05-19

    Moisture content (MC) is one of the most important quality parameters of green coffee beans. Therefore, its fast and reliable measurement is necessary. This study evaluated the feasibility of near infrared (NIR) spectroscopy and chemometrics for rapid and non-destructive prediction of MC in intact green coffee beans of both Coffeaarabica (Arabica) and Coffeacanephora (Robusta) species. Diffuse reflectance (log 1/R) spectra of intact beans were acquired using a bench top Fourier transform NIR instrument. MC was determined gravimetrically according to The International Organization for Standardization (ISO) 6673. Samples were split into subsets for calibration (n = 64) and independent validation (n = 44). A three-component partial least squares regression (PLSR) model using raw NIR spectra yielded a root mean square error of prediction (RMSEP) of 0.80% MC; a four component PLSR model using scatter corrected spectra yielded a RMSEP of 0.57% MC. A simplified PLS model using seven selected wavelengths (1155, 1212, 1340, 1409, 1724, 1908, and 2249 nm) yielded a similar accuracy (RMSEP: 0.77% MC) which opens the possibility of creating cheaper NIR instruments. In conclusion, NIR diffuse reflectance spectroscopy appears to be suitable for rapid and reliable MC prediction in intact green coffee; no separate model for Arabica and Robusta species is needed.

  15. The intact structural form of LLO in endosomes cannot protect against listeriosis

    Science.gov (United States)

    Rodriguez-Del Rio, Estela; Frande-Cabanes, Elisabet; Tobes, Raquel; Pareja, Eduardo; Lecea-Cuello, M. Jesús; Ruiz-Sáez, Marta; Carrasco-Marín, Eugenio; Alvarez-Dominguez, Carmen

    2011-01-01

    LLO is the major immuno-dominant antigen in listeriosis and is also required for protective immunity. Two forms of LLO can be observed in endosomal membranes, a LLO intact form and a Ctsd-processed LLO1-491 form. Endosomes obtained from resting macrophages contained only LLO intact forms, while endosomes obtained from IFN-activated macrophages contained both forms. Both types of endosomes elicited LLO90-91/CD8+ and LLO189-201/CD4+ specific immune responses. However, only endosomes containing the Ctsd-processed LLO1-491 form showed significant CD4+ and CD8+ T cell responses similar to LM infected bone marrow derived macrophages and characteristic of protective Listeria immunity. Moreover, endosomes with intact LLO could not confer protection as vaccine carriers against murine listeriosis. While endosomes with Ctsd-processed LLO1-491 form showed a moderate ability, slightly lower than high efficiency vaccine vectors as MØ infected with LM. These studies argue that all cell-free membrane vesicles might serve as valid vaccine carriers against infectious agents. Exclusively those cell-free vesicles MIIC competent for LLO processing are protective vaccines vectors since they recruit significant numbers of mature dendritic cells to the vaccination sites and contain a LLO1-491 form that might be accessible for MHC class I and class II antigen presentation. PMID:22003433

  16. Microchip capillary electrophoresis-electrospray ionization-mass spectrometry of intact proteins using uncoated Ormocomp microchips.

    Science.gov (United States)

    Sikanen, Tiina; Aura, Susanna; Franssila, Sami; Kotiaho, Tapio; Kostiainen, Risto

    2012-01-20

    We present rapid (microchips. The microchips are fabricated fully of commercial inorganic-organic hybrid material, Ormocomp, by UV-embossing and adhesive Ormocomp-Ormocomp bonding (CE microchannels). A sheath-flow ESI interface is monolithically integrated with the UV-embossed separation channels by cutting a rectangular emitter tip in the end with a dicing saw. As a result, electrospray was produced from the corner of chip with good reproducibility between parallel tips (stability within 3.8-9.2% RSD). Thanks to its inherent biocompatibility and stable (negative) surface charge, Ormocomp microchips enable efficient intact protein analysis with up to ∼10(4) theoretical separation plates per meter without any chemical or physical surface modification before analysis. The same microchip setup is also feasible for rapid peptide sequencing and mass fingerprinting and shows excellent migration time repeatability from run to run for both peptides (5.6-5.9% RSD, n=4) and intact proteins (1.3-7.5% RSD, n=3). Thus, the Ormocomp microchips provide a versatile new tool for MS-based proteomics. Particularly, the feasibility of the Ormocomp chips for rapid analysis of intact proteins with such a simple setup is a valuable increment to the current technology.

  17. The effect of self-transcendence on depression in cognitively intact nursing home patients.

    Science.gov (United States)

    Haugan, Gørill; Innstrand, Siw Tone

    2012-01-01

    Aims. This study's aim was to test the effects of self-transcendence on depression among cognitively intact nursing home patients. Background. Depression is considered the most frequent mental disorder among the elderly population. Specifically, the depression rate among nursing home patients is three to four times higher than that among community-dwelling elderly. Therefore, finding new and alternative ways to prevent and decrease depression is of great importance for nursing home patients' well-being. Self-transcendence is related to spiritual as well as nonspiritual factors, and it is described as a correlate and resource for well-being among vulnerable populations and at the end of life. Methods. A two-factor construct of the self-transcendence scale (interpersonal and intrapersonal) and the hospital anxiety and depression scale (HADS) was applied. A sample of 202 cognitively intact nursing home patients in central Norway was selected to respond to the questionnaires in 2008/2009. Results. A hypothesized SEM model demonstrated significant direct relationships and total effects of self-transcendence on depression. Conclusion and Implication for Practice. Facilitating patients' self-transcendence, both interpersonally and intrapersonally, might decrease depression among cognitively intact nursing home patients.

  18. Crosstalk in the mouse thymus

    NARCIS (Netherlands)

    W. van Ewijk (Willem); E.W. Shores (Elisabeth); A. Singer (Andrew)

    1994-01-01

    textabstractThe development of mature T cells within the thymus is dependent upon intact cortical and medullary microenvironments. In turn, thymic microenvironment themselves are dependent on lymphoid cells to maintain their integrity. Here, Willem van Ewijk and colleagues discuss experiments that h

  19. NCBI nr-aa BLAST: CBRC-STRI-01-2687 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-2687 ref|NP_444455.1| vomeronasal 1 receptor, B1 [Mus musculus] sp|Q9E...P51|VN1B1_MOUSE RecName: Full=Vomeronasal type-1 receptor B1; AltName: Full=Vomeronasal type-1 receptor A5; AltName: Full=Vomeronasa...l receptor 2; AltName: Full=Pheromone receptor VN2 gb|AAG42083.1|AF291489_1 vomeronasa...l receptor V1RB1 [Mus musculus] gb|AAG43248.1| VN2 [Mus musculus] gb|AAI07184.1| Vomeronasa...l 1 receptor, B1 [Mus musculus] gb|EDK99276.1| vomeronasal 1 receptor, B1 [Mus musculus] NP_444455.1 2e-30 35% ...

  20. Multi-Axis Prosthetic Knee Resembles Alpine Skiing Movements of an Intact Leg

    Directory of Open Access Journals (Sweden)

    Ivan Demšar, Jože Duhovnik, Blaž Lešnik, Matej Supej

    2015-12-01

    Full Text Available The purpose of the study was to analyse the flexion angles of the ski boot, ankle and knee joints of an above-knee prosthesis and to compare them with an intact leg and a control group of skiers. One subject with an above-knee amputation of the right leg and eight healthy subjects simulated the movement of a skiing turn by performing two-leg squats in laboratory conditions. By adding additional loads in proportion to body weight (BW; +1/3 BW, +2/3 BW, +3/3 BW, various skiing regimes were simulated. Change of Flexion Angle (CoFA and Range of Motion (RoM in the ski boot, ankle and knee joints were calculated and compared. An average RoM in the skiing boot on the side of prosthesis (4.4 ± 1.1° was significantly lower compared to an intact leg (5.9 ± 1.8° and the control group (6.5 ± 2.3°. In the ankle joint, the average RoM was determined to be 13.2±2.9° in the prosthesis, 12.7 ± 2.8° in an intact leg and 14.8±3.6 in the control group. However, the RoM of the knee joint in the prosthesis (42.2 ± 4.2° was significantly larger than that of the intact leg (34.7 ± 4.4°. The average RoM of the knee joint in the control group was 47.8 ± 5.4°. The influences of additional loads on the kinematics of the lower extremities were different on the side of the prosthesis and on the intact leg. In contrast, additional loads did not produce any significant differences in the control group. Although different CoFAs in the ski boot, ankle and knee joints were used, an above-knee prosthesis with a built-in multi-axis prosthetic knee enables comparable leg kinematics in simulated alpine skiing.

  1. Molecular properties of adult mouse gastric and intestinal epithelial progenitors in their niches

    DEFF Research Database (Denmark)

    Giannakis, Marios; Stappenbeck, Thaddeus S; Mills, Jason C

    2006-01-01

    We have sequenced 36,641 expressed sequence tags from laser capture microdissected adult mouse gastric and small intestinal epithelial progenitors, obtaining 4031 and 3324 unique transcripts, respectively. Using Gene Ontology (GO) terms, each data set was compared with cDNA libraries from intact...... adult stomach and small intestine. Genes in GO categories enriched in progenitors were filtered against genes in GO categories represented in hematopoietic, neural, and embryonic stem cell transcriptomes and mapped onto transcription factor networks, plus canonical signal transduction and metabolic...

  2. Optical coherence elastography for cellular-scale stiffness imaging of mouse aorta

    Science.gov (United States)

    Wijesinghe, Philip; Johansen, Niloufer J.; Curatolo, Andrea; Sampson, David D.; Ganss, Ruth; Kennedy, Brendan F.

    2017-04-01

    We have developed a high-resolution optical coherence elastography system capable of estimating Young's modulus in tissue volumes with an isotropic resolution of 15 μm over a 1 mm lateral field of view and a 100 μm axial depth of field. We demonstrate our technique on healthy and hypertensive, freshly excised and intact mouse aortas. Our technique has the capacity to delineate the individual mechanics of elastic lamellae and vascular smooth muscle. Further, we observe global and regional vascular stiffening in hypertensive aortas, and note the presence of local micro-mechanical signatures, characteristic of fibrous and lipid-rich regions.

  3. Mouse models for cancer research

    Institute of Scientific and Technical Information of China (English)

    Wei Zhang; Lynette Moore; Ping Ji

    2011-01-01

    Mouse models of cancer enable researchers to leamn about tumor biology in complicated and dynamic physiological systems. Since the development of gene targeting in mice, cancer biologists have been among the most frequent users of transgenic mouse models, which have dramatically increased knowledge about how cancers form and grow. The Chinese Joumnal of Cancer will publish a series of papers reporting the use of mouse models in studying genetic events in cancer cases. This editorial is an overview of the development and applications of mouse models of cancer and directs the reader to upcoming papers describing the use of these models to be published in coming issues, beginning with three articles in the current issue.

  4. Computer Workstation: Pointer/Mouse

    Science.gov (United States)

    ... Safety and Health Program Recommendations It's the Law Poster REGULATIONS Law and Regulations Standard Interpretations Training Requirements ... when evaluating your computer workstation. Pointer Placement Pointer Size, Shape, and Settings Pointer/Mouse Quick Tips Keep ...

  5. The second face of blindness: processing speed deficits in the intact visual field after pre- and post-chiasmatic lesions.

    Directory of Open Access Journals (Sweden)

    Michał Bola

    Full Text Available PURPOSE: Damage along the visual pathway results in a visual field defect (scotoma, which retinotopically corresponds to the damaged neural tissue. Other parts of the visual field, processed by the uninjured tissue, are considered to be intact. However, perceptual deficits have been observed in the "intact" visual field, but these functional impairments are poorly understood. We now studied temporal processing deficits in the intact visual field of patients with either pre- or post-chiasmatic lesions to better understand the functional consequences of partial blindness. METHODS: Patients with pre- (n = 53 or post-chiasmatic lesions (n = 98 were tested with high resolution perimetry--a method used to map visual fields with supra-threshold light stimuli. Reaction time of detections in the intact visual field was then analyzed as an indicator of processing speed and correlated with features of the visual field defect. RESULTS: Patients from both groups exhibited processing speed deficits in their presumably "intact" field as indicated by comparison to a normative sample. Further, in both groups processing speed was found to be a function of two factors. Firstly, a spatially restricted (retinotopic influence of the scotoma was seen in longer reaction times when stimuli were presented in intact field sectors close to the defect. Secondly, patients with larger scotomata had on average longer reaction times in their intact field indicating a more general (non-retinotopic influence of the scotoma. CONCLUSIONS: Processing speed deficits in the "intact" visual field of patients with visual system damage demonstrate that visual system lesions have more widespread consequences on perception than previously thought. Because dysfunctions of the seeing field are expected to contribute to subjective vision, including visual tests of the presumed "intact" field may help to better understand vision loss and to improve methods of vision restoration and

  6. Hand gestures mouse cursor control

    Directory of Open Access Journals (Sweden)

    Marian-Avram Vincze

    2014-05-01

    Full Text Available The paper describes the implementation of a human-computer interface for controlling the mouse cursor. The test reveal the fact: a low-cost web camera some processing algorithms are quite enough to control the mouse cursor on computers. Even if the system is influenced by the illuminance level on the plane of the hand, the current study may represent a start point for some studies on the hand tracking and gesture recognition field.

  7. Somatostatin Interneurons Control a Key Component of Mismatch Negativity in Mouse Visual Cortex

    Directory of Open Access Journals (Sweden)

    Jordan P. Hamm

    2016-07-01

    Full Text Available Patients with schizophrenia have deficient sensory processing, undermining how they perceive and relate to a changing environment. This impairment can be captured by the reduced mismatch negativity (MMN index, an electroencephalographic biomarker of psychosis. The biological factors contributing to MMN are unclear, though mouse research, in which genetic and optical methods could be applied, has given some insight. Using fast two-photon calcium imaging and multielectrode recordings in awake mice, we find that visual cortical circuits display adapted (decreased responses to repeated stimuli and amplified responses to a deviant stimulus, the key component of human MMN. Moreover, pharmacogenetic silencing of somatostatin-containing interneurons specifically eliminated this amplification, along with its associated theta/alpha-band response, leaving stimulus-specific adaption and related gamma-band modulations intact. Our results validate a mouse model of MMN and suggest that abnormalities in somatostatin-containing interneurons cause sensory deficits underlying MMN and schizophrenia.

  8. Oxygen sensing and conducted vasomotor responses in mouse cremaster arterioles in situ

    DEFF Research Database (Denmark)

    Ngo, Thuc Anh; Jensen, Lars Jørn; Riemann, Mads Achen

    2010-01-01

    This study examines mechanisms by which changes in tissue oxygen tension elicit vasomotor responses and whether localized changes in oxygen tension initiates conducted vasomotor responses in mouse cremaster arterioles. Intravital microscopy was used to visualize the mouse cremaster microcirculation....... The cremaster was superfused with Krebs' solution with different oxygen tensions, and a gas exchange chamber was used to induce localized changes in oxygen tension. In arterioles where red blood cells were removed by buffer perfusion, arterioles responded with same magnitudes of vasodilatation (DeltaD = 16.......0 +/- 4.9 mum) when changing from high (PO(2) = 242.5 +/- 13.3 mm Hg) to low (PO(2) = 22.5 +/- 4.8 mm Hg) oxygen tension as seen in the intact cremaster circulation (DeltaD = 18.7 +/- 1.0 mum). Blockade of NO synthases by L: -NAME and adenosine receptors by DPCPX had no effects on vasomotor responses...

  9. Specific amplification by PCR of rearranged genomic variable regions of immunoglobulin genes from mouse hybridoma cells.

    Science.gov (United States)

    Berdoz, J; Monath, T P; Kraehenbuhl, J P

    1995-04-01

    We have designed a novel strategy for the isolation of the rearranged genomic fragments encoding the L-VH-D-JH and L-V kappa/lambda-J kappa/lambda regions of mouse immunoglobulin genes. This strategy is based on the PCR amplification of genomic DNA from mouse hybridomas using multiple specific primers chosen in the 5'-untranslated region and in the intron downstream of the rearranged JH/J kappa/lambda sequences. Variable regions with intact coding sequences, including full-length leader peptides (L) can be obtained without previous DNA sequencing. Our strategy is based on a genomic template that produces fragments that do not need to be adapted for recombinant antibody expression, thus facilitating the generation of chimeric and isotype-switched immunoglobulins.

  10. Mouse models of dengue virus infection for vaccine testing.

    Science.gov (United States)

    Sarathy, Vanessa V; Milligan, Gregg N; Bourne, Nigel; Barrett, Alan D T

    2015-12-10

    Dengue is a mosquito-borne disease caused by four serologically and genetically related viruses termed DENV-1 to DENV-4. With an annual global burden of approximately 390 million infections occurring in the tropics and subtropics worldwide, an effective vaccine to combat dengue is urgently needed. Historically, a major impediment to dengue research has been development of a suitable small animal infection model that mimics the features of human illness in the absence of neurologic disease that was the hallmark of earlier mouse models. Recent advances in immunocompromised murine infection models have resulted in development of lethal DENV-2, DENV-3 and DENV-4 models in AG129 mice that are deficient in both the interferon-α/β receptor (IFN-α/β R) and the interferon-γ receptor (IFN-γR). These models mimic many hallmark features of dengue disease in humans, such as viremia, thrombocytopenia, vascular leakage, and cytokine storm. Importantly AG129 mice develop lethal, acute, disseminated infection with systemic viral loads, which is characteristic of typical dengue illness. Infected AG129 mice generate an antibody response to DENV, and antibody-dependent enhancement (ADE) models have been established by both passive and maternal transfer of DENV-immune sera. Several steps have been taken to refine DENV mouse models. Viruses generated by peripheral in vivo passages incur substitutions that provide a virulent phenotype using smaller inocula. Because IFN signaling has a major role in immunity to DENV, mice that generate a cellular immune response are desired, but striking the balance between susceptibility to DENV and intact immunity is complicated. Great strides have been made using single-deficient IFN-α/βR mice for DENV-2 infection, and conditional knockdowns may offer additional approaches to provide a panoramic view that includes viral virulence and host immunity. Ultimately, the DENV AG129 mouse models result in reproducible lethality and offer multiple

  11. Mechanical analysis of the human cadaveric thoracic spine with intact rib cage.

    Science.gov (United States)

    Mannen, Erin M; Anderson, John T; Arnold, Paul M; Friis, Elizabeth A

    2015-07-16

    The goal of this study was to characterize the overall in-plane and basic coupled motion of a cadaveric human thoracic spine with intact true ribs. Researchers are becoming increasingly interested in the thoracic spine due to both the high prevalence of injury and pain in the region and also innovative surgical techniques that utilize the rib cage. Computational models can be useful tools to predict loading patterns and understand effects of surgical procedures or medical devices, but they are often limited by insufficient cadaveric input data. In this study, pure moments to ±5 Nm were applied in flexion-extension, lateral bending, and axial rotation to seven human cadaveric thoracic spine specimens (T1-T12) with intact true ribs to determine symmetry of in-plane motion, differences in neutral and elastic zone motion and stiffness, and significance of out-of-plane rotations and translations. Results showed that lateral bending and axial rotation exhibited symmetric motion, neutral and elastic zone motion and stiffness values were significantly different for all modes of bending (p<0.05), and out-of-plane rotations and translations were greater than zero for most rotations and translations. Overall in-plane rotations were 7.7±3.4° in flexion, 9.6±3.7° in extension, 23.3±8.4° in lateral bending, and 26.3±12.2° in axial rotation. Results of this study could provide inputs or validation comparisons for computational models. Future studies should characterize coupled motion patterns and local and regional level biomechanics of cadaveric human thoracic spines with intact true ribs.

  12. Human skin penetration of silver nanoparticles through intact and damaged skin.

    Science.gov (United States)

    Larese, Francesca Filon; D'Agostin, Flavia; Crosera, Matteo; Adami, Gianpiero; Renzi, Nadia; Bovenzi, Massimo; Maina, Giovanni

    2009-01-01

    There is a growing interest on nanoparticle safety for topical use. The benefits of nanoparticles have been shown in several scientific fields, but little is known about their potential to penetrate the skin. This study aims at evaluating in vitro skin penetration of silver nanoparticles. Experiments were performed using the Franz diffusion cell method with intact and damaged human skin. Physiological solution was used as receiving phase and 70 microg/cm2 of silver nanoparticles coated with polyvinylpirrolidone dispersed in synthetic sweat were applied as donor phase to the outer surface of the skin for 24h. The receptor fluid measurements were performed by electro thermal atomic absorption spectroscopy (ETAAS). Human skin penetration was also determined by using transmission electron microscope (TEM) to verify the location of silver nanoparticles in exposed membranes. Median silver concentrations of 0.46 ng cm(-2) (range intact skin (eight cells) and on damaged skin (eight cells), respectively. Twenty-four hours silver flux permeation in damaged skin was 0.62+/-0.2 ng cm(-2) with a lag time <1h. Our experimental data showed that silver nanoparticles absorption through intact and damaged skin was very low but detectable, and that in case of damaged skin it was possible an increasing permeation of silver applied as nanoparticles. Moreover, silver nanoparticles could be detected in the stratum corneum and the outermost surface of the epidermis by electron microscopy. We demonstrated for the first time that silver applied as nanoparticles coated with polyvinylpirrolidone is able to permeate the damaged skin in an in vitro diffusion cell system.

  13. Human skin penetration of gold nanoparticles through intact and damaged skin.

    Science.gov (United States)

    Filon, Francesca Larese; Crosera, Matteo; Adami, Gianpiero; Bovenzi, Massimo; Rossi, Federica; Maina, Giovanni

    2011-12-01

    Gold nanoparticles (AuNPs) are produced for many applications but there is a lack of available data on their skin absorption. Experiments were performed using the Franz diffusion cell method with intact and damaged human skin. A physiological solution was used as receiving phase and 0.5 mL (1st exp) and 1.5 mL (2nd exp) of a solution containing 100 mg L⁻¹ of AuNPs (15 and 45 μg cm⁻², respectively) was applied as donor phase to the outer surface of the skin for 24 h. Skin absorption was dose dependent. Mean gold content of 214.0 ± 43.7 ng cm⁻² and 187.7 ± 50.2 ng cm⁻² were found in the receiving solutions of cells where the AuNPs solution was applied in higher concentration on intact skin (8 Franz cells) and on damaged skin (8 Franz cells), respectively. Twenty-four hours gold flux permeation was 7.8 ± 2.0 ng cm⁻² h⁻¹ and 7.1 ± 2.5 ng cm⁻² h⁻¹ in intact and damaged skin, respectively, with a lag time less than 1 hour. Transmission Electron Microscope analysis on skin samples and chemical analysis using Inductively Coupled Plasma-Mass Spectrometry demonstrated the presence of AuNPs into epidermis and dermis. This study showed that AuNPs are able to penetrate the human skin in an in vitro diffusion cell system.

  14. Mechanisms of Granule Membrane Recapture following Exocytosis in Intact Mast Cells*

    Science.gov (United States)

    Cabeza, Jose M.; Acosta, Jorge; Alés, Eva

    2013-01-01

    In secretory cells, several exocytosis-coupled forms of endocytosis have been proposed including clathrin-mediated endocytosis, kiss-and-run endocytosis, cavicapture, and bulk endocytosis. These forms of endocytosis can be induced under different conditions, but their detailed molecular mechanisms and functions are largely unknown. We studied exocytosis and endocytosis in mast cells with both perforated-patch and whole-cell configurations of the patch clamp technique using cell capacitance measurements in combination with amperometric serotonin detection. We found that intact mast cells exhibit an early endocytosis that follows exocytosis induced by compound 48/80. Direct observation of individual exocytic and endocytic events showed a higher percentage of capacitance flickers (27.3%) and off-steps (11.4%) in intact mast cells than in dialyzed cells (5.4% and 2.9%, respectively). Moreover, we observed a type of endocytosis of large pieces of membrane that were likely formed by cumulative fusion of several secretory granules with the cell membrane. We also identified “large-capacitance flickers” that occur after large endocytosis events. Pore conductance analysis indicated that these transient events may represent “compound cavicapture,” most likely due to the flickering of a dilated fusion pore. Using fluorescence imaging of individual exocytic and endocytic events we observed that granules can fuse to granules already fused with the plasma membrane, and then the membranes and dense cores of fused granules are internalized. Altogether, our results suggest that stimulated exocytosis in intact mast cells is followed by several forms of compensatory endocytosis, including kiss-and-run endocytosis and a mechanism for efficient retrieval of the compound membrane of several secretory granules through a single membrane fission event. PMID:23709219

  15. Analysis of responses to glyceryl trinitrate and sodium nitrite in the intact chest rat.

    Science.gov (United States)

    Nossaman, Bobby D; Pankey, Edward A; Badejo, Adeleke R; Casey, David B; Uppu, Satvika; Murthy, Subramanyam N; Kadowitz, Philip J

    2012-05-15

    Responses to glyceryl trinitrate/nitroglycerin (GTN), S-nitrosoglutathione (GSNO), and sodium nitrite were compared in the intact chest rat. The iv injections of GTN, sodium nitrite, and GSNO produced dose-dependent decreases in pulmonary and systemic arterial pressures. In as much as cardiac output was not reduced, the decreases in pulmonary and systemic arterial pressures indicate that GTN, sodium nitrite, and GSNO have significant vasodilator activity in the pulmonary and systemic vascular beds in the rat. Responses to GTN were attenuated by cyanamide, but not allopurinol, whereas responses to nitrite formed by the metabolism of GTN were attenuated by allopurinol and cyanamide. The results with allopurinol and cyanamide suggest that only mitochondrial aldehyde dehydrogenase is involved in the bioactivation of GTN, sodium nitrite, and GSNO, whereas both pathways are involved in the bioactivation of nitrite anion in the intact rat. The comparison of vasodilator activity indicates that GSNO and GTN are more than 1000-fold more potent than sodium nitrite in decreasing pulmonary and systemic arterial pressures in the rat. Following administration of 1H-[1,2,4]-oxadizaolo[4,3-]quinoxaline-1-one (ODQ), responses to GTN were significantly attenuated, indicating that responses are mediated by the activation of soluble guanylyl cyclase. These data suggest that the reduction of nitrite to nitric oxide formed from the metabolism of GTN, cannot account for the vasodilator activity of GTN in the intact rat and that another mechanism; perhaps the formation of an S-NO, may mediate the vasodilator response to GTN in this species.

  16. Sediment properties and CO2 efflux from intact and cleared temperate mangrove forests

    Science.gov (United States)

    Bulmer, R. H.; Lundquist, C. J.; Schwendenmann, L.

    2015-10-01

    Temperate mangrove forests in New Zealand have increased in area over recent decades. Expansion of temperate mangroves in New Zealand is associated with perceived loss of other estuarine habitats, and decreased recreational and amenity values, resulting in clearing of mangrove forests. In the tropics, changes in sediment characteristics and carbon efflux have been reported following mangrove clearance. This is the first study in temperate mangrove (Avicennia marina) forests investigating the impact of clearing on sediment CO2 efflux and associated biotic and abiotic factors. Sediment CO2 efflux rates from intact (168.5 ± 45.8 mmol m-2 d-1) and cleared (133.9 ± 37.2 mmol m-2 d-1) mangrove forests in New Zealand are comparable to rates measured in tropical mangrove forests. We did not find a significant difference in sediment CO2 efflux rates between intact and cleared temperate mangrove forests. Pre-shading the sediment for more than 30 min prior to dark chamber measurements was found to have no significant effect on sediment CO2 efflux. This suggests that the continuation of photosynthetic CO2 uptake by biofilm communities was not occurring after placement of dark chambers. Rather, above-ground mangrove biomass, sediment temperature and chlorophyll a concentration were the main factors explaining the variability in sediment CO2 efflux in intact mangrove forests. The main factors influencing sediment CO2 efflux in cleared mangrove forest sites were sediment organic carbon concentration, nitrogen concentration and sediment grain size. Our results show that greater consideration should be given regarding the rate of carbon released from mangrove forest following clearance and the relative contribution to global carbon emissions.

  17. An intact dorsomedial posterior arcuate nucleus is not necessary for photoperiodic responses in Siberian hamsters.

    Science.gov (United States)

    Teubner, Brett J W; Leitner, Claudia; Thomas, Michael A; Ryu, Vitaly; Bartness, Timothy J

    2015-04-01

    Seasonal responses of many animal species are triggered by changes in daylength and its transduction into a neuroendocrine signal by the pineal gland through the nocturnal duration of melatonin (MEL) release. The precise central sites necessary to receive, transduce, and relay the short day (SD) fall-winter MEL signals into seasonal responses and changes in physiology and behavior are unclear. In Siberian hamsters, SDs trigger decreases in body and lipid mass, testicular regression and pelage color changes. Several candidate genes and their central sites of expression have been proposed as components of the MEL transduction system with considerable recent focus on the arcuate nucleus (ARC) and its component, the dorsomedial posterior arcuate nucleus (dmpARC). This site has been postulated as a critical relay of SD information through the modulation of a variety of neurochemicals/receptors important for the control of energy balance. Here the necessity of an intact dmpARC for SD responses was tested by making electrolytic lesions of the Siberian hamster dmpARC and then exposing them to either long days (LD) or SDs for 12wks. The SD typical decreases in body and fat mass, food intake, testicular volume, serum testosterone concentrations, pelage color change and increased UCP-1 protein expression (a proxy for brown adipose tissue thermogenesis) all occurred despite the lack of an intact dmpARC. Although the Siberian hamster dmpARC contains photoperiod-modulated constituents, these data demonstrate that an intact dmpARC is not necessary for SD responses and not integral to the seasonal energy- and reproductive-related responses measured here.

  18. Palladium nanoparticles exposure: Evaluation of permeation through damaged and intact human skin.

    Science.gov (United States)

    Larese Filon, Francesca; Crosera, Matteo; Mauro, Marcella; Baracchini, Elena; Bovenzi, Massimo; Montini, Tiziano; Fornasiero, Paolo; Adami, Gianpiero

    2016-07-01

    The intensified use of palladium nanoparticles (PdNPs) in many chemical reactions, jewellery, electronic devices, in car catalytic converters and in biomedical applications lead to a significant increase in palladium exposure. Pd can cause allergic contact dermatitis when in contact with the skin. However, there is still a lack of toxicological data related to nano-structured palladium and information on human cutaneous absorption. In fact, PdNPs, can be absorbed through the skin in higher amounts than bulk Pd because NPs can release more ions. In our study, we evaluated the absorption of PdNPs, with a size of 10.7 ± 2.8 nm, using intact and damaged human skin in Franz cells. 0.60 mg cm(-2) of PdNPs were applied on skin surface for 24 h. Pd concentrations in the receiving solutions at the end of experiments were 0.098 ± 0.067 μg cm(-2) and 1.06 ± 0.44 μg cm(-2) in intact skin and damaged skin, respectively. Pd flux permeation after 24 h was 0.005 ± 0.003 μg cm(-2) h(-1) and 0.057 ± 0.030 μg cm(-2) h(-1) and lag time 4.8 ± 1.7 and 4.2 ± 3.6 h, for intact and damaged skin respectively. This study indicates that Pd can penetrate human skin.

  19. Correlations Between Physical and Hydraulic Properties and Uranium Desorption in Contaminated, Intact Sediment Cores

    Science.gov (United States)

    Rockhold, M. L.; Oostrom, M.; Wietsma, T. W.; Zachara, J. M.

    2010-12-01

    An unlined disposal pond in the 300 Area of the Hanford Site received uranium-bearing liquid effluents associated with nuclear reactor fuel rod processing from 1943 to 1975. Contaminated sediments from the base and sides of the former pond were excavated and removed from the site in the early 1990s, but a uranium plume has persisted in the groundwater at concentrations exceeding the drinking water standard. The former process pond is located adjacent to the Columbia River and seasonal fluctuations in the river stage and water table provide a mechanism for resupplying residual uranium from the vadose zone to the groundwater when the lower vadose zone is periodically rewetted. Intact cores were collected from the site for measurements of physical, hydraulic, and geochemical properties. Multistep outflow experiments were also performed on the intact cores to determine permeability-saturation-capillary pressure relations. Pore water displaced during these experiments for two of the vadose zone cores was also analyzed for uranium. For a core containing finer-textured sediment classified as muddy sandy gravel, and a core containing coarser-textured sediment classified as gravel, the relative aqueous uranium concentrations increased by factors of 8.3 and 1.5, respectively, as the cores were desaturated and progressively smaller pore-size classes were drained. Aqueous concentrations of uranium in the extracted pore waters were up to 115 times higher than the current drinking water standard of 30 ppb. These results confirm that there is a continuing source of uranium in the vadose zone at the site, and are consistent with a hypothesis that the persistence of the groundwater uranium plume is also associated, in part, with rate-limited mass transfer from finer-textured sediments. The data from these and several other intact cores from the site are evaluated to explore relationships between physical and hydraulic properties and uranium desorption characteristics.

  20. Design and characterisation of a phased antenna array for intact breast hyperthermia.

    Science.gov (United States)

    Curto, Sergio; Garcia-Miquel, Aleix; Suh, Minyoung; Vidal, Neus; Lopez-Villegas, Jose M; Prakash, Punit

    2017-06-28

    Currently available hyperthermia technology is not well suited to treating cancer malignancies in the intact breast. This study investigates a microwave applicator incorporating multiple patch antennas, with the goal of facilitating controllable power deposition profiles for treating lesions at diverse locations within the intact breast. A 3D-computational model was implemented to assess power deposition profiles with 915 MHz applicators incorporating a hemispheric groundplane and configurations of 2, 4, 8, 12, 16 and 20 antennas. Hemispheric breast models of 90 mm and 150 mm diameter were considered, where cuboid target volumes of 10 mm edge length (1 cm(3)) and 30 mm edge length (27 cm(3)) were positioned at the centre of the breast, and also located 15 mm from the chest wall. The average power absorption (αPA) ratio expressed as the ratio of the PA in the target volume and in the full breast was evaluated. A 4-antenna proof-of-concept array was fabricated and experimentally evaluated. Computational models identified an optimal inter-antenna spacing of 22.5° along the applicator circumference. Applicators with 8 and 12 antennas excited with constant phase presented the highest αPA at centrally located and deep-seated targets, respectively. Experimental measurements with a 4-antenna proof-of-concept array illustrated the potential for electrically steering power deposition profiles by adjusting the relative phase of the signal at antenna inputs. Computational models and experimental results suggest that the proposed applicator may have potential for delivering conformal thermal therapy in the intact breast.

  1. Bioengineered human IAS reconstructs with functional and molecular properties similar to intact IAS.

    Science.gov (United States)

    Singh, Jagmohan; Rattan, Satish

    2012-09-15

    Because of its critical importance in rectoanal incontinence, we determined the feasibility to reconstruct internal anal sphincter (IAS) from human IAS smooth muscle cells (SMCs) with functional and molecular attributes similar to the intact sphincter. The reconstructs were developed using SMCs from the circular smooth muscle layer of the human IAS, grown in smooth muscle differentiation media under sterile conditions in Sylgard-coated tissue culture plates with central Sylgard posts. The basal tone in the reconstructs and its changes were recorded following 0 Ca(2+), KCl, bethanechol, isoproterenol, protein kinase C (PKC) activator phorbol 12,13-dibutyrate, and Rho kinase (ROCK) and PKC inhibitors Y-27632 and Gö-6850, respectively. Western blot (WB), immunofluorescence (IF), and immunocytochemical (IC) analyses were also performed. The reconstructs developed spontaneous tone (0.68 ± 0.26 mN). Bethanechol (a muscarinic agonist) and K(+) depolarization produced contraction, whereas isoproterenol (β-adrenoceptor agonist) and Y-27632 produced a concentration-dependent decrease in the tone. Maximal decrease in basal tone with Y-27632 and Gö-6850 (each 10(-5) M) was 80.45 ± 3.29 and 17.76 ± 3.50%, respectively. WB data with the IAS constructs' SMCs revealed higher levels of RhoA/ROCK, protein kinase C-potentiated inhibitor or inhibitory phosphoprotein for myosin phosphatase (CPI-17), phospho-CPI-17, MYPT1, and 20-kDa myosin light chain vs. rectal smooth muscle. WB, IF, and IC studies of original SMCs and redispersed from the reconstructs for the relative distribution of different signal transduction proteins confirmed the feasibility of reconstruction of IAS with functional properties similar to intact IAS and demonstrated the development of myogenic tone with critical dependence on RhoA/ROCK. We conclude that it is feasible to bioengineer IAS constructs using human IAS SMCs that behave like intact IAS.

  2. Impact of extreme weather on critical infrastructure: the EU-INTACT risk framework

    Directory of Open Access Journals (Sweden)

    Tagg Andrew

    2016-01-01

    Full Text Available Resilience of critical infrastructure (CI to extreme weather events, such as heavy rainfall, high temperatures and winter storms, is one of the most demanding challenges for governments and society. Recent experiences have highlighted the economic and societal reliance on a dependable and resilient infrastructure, and the far-reaching impacts that outages or malfunctions can have. Growing scientific evidence indicates that more severe and frequent extreme weather events are likely. The EU-funded INTACT project addresses these CI challenges and attempts to bring together cutting-edge knowledge and experience from across Europe to inform the development of best practice approaches in planning, crisis response and recovery capabilities. The project considers the options for mitigating the extreme weather impacts. A key component of the INTACT project is the development of a risk management structure to support decision-making in the case studies. This structure forms part of the overall INTACT Wiki: the main output of the project. It comprises a risk ‘framework’ that sets out how information and guidance can be accessed by CI owners and operators. Within this there is a step-wise risk assessment process based on best practice from the IEC. The risk framework and process presents: structures for models and data requirements for decision making; identifies tools and methods that support decision making; supports analysis of measures to protect CI through simulation; and indicates gaps in modelling and data availability. This paper outlines the components of the risk framework and process, and illustrates its use in a case study dealing with electricity supply and winter storms.

  3. Prototypic and Arkypallidal Neurons in the Dopamine-Intact External Globus Pallidus

    Science.gov (United States)

    Abdi, Azzedine; Mallet, Nicolas; Mohamed, Foad Y.; Sharott, Andrew; Dodson, Paul D.; Nakamura, Kouichi C.; Suri, Sana; Avery, Sophie V.; Larvin, Joseph T.; Garas, Farid N.; Garas, Shady N.; Vinciati, Federica; Morin, Stéphanie; Bezard, Erwan

    2015-01-01

    Studies in dopamine-depleted rats indicate that the external globus pallidus (GPe) contains two main types of GABAergic projection cell; so-called “prototypic” and “arkypallidal” neurons. Here, we used correlative anatomical and electrophysiological approaches in rats to determine whether and how this dichotomous organization applies to the dopamine-intact GPe. Prototypic neurons coexpressed the transcription factors Nkx2-1 and Lhx6, comprised approximately two-thirds of all GPe neurons, and were the major GPe cell type innervating the subthalamic nucleus (STN). In contrast, arkypallidal neurons expressed the transcription factor FoxP2, constituted just over one-fourth of GPe neurons, and innervated the striatum but not STN. In anesthetized dopamine-intact rats, molecularly identified prototypic neurons fired at relatively high rates and with high regularity, regardless of brain state (slow-wave activity or spontaneous activation). On average, arkypallidal neurons fired at lower rates and regularities than prototypic neurons, and the two cell types could be further distinguished by the temporal coupling of their firing to ongoing cortical oscillations. Complementing the activity differences observed in vivo, the autonomous firing of identified arkypallidal neurons in vitro was slower and more variable than that of prototypic neurons, which tallied with arkypallidal neurons displaying lower amplitudes of a “persistent” sodium current important for such pacemaking. Arkypallidal neurons also exhibited weaker driven and rebound firing compared with prototypic neurons. In conclusion, our data support the concept that a dichotomous functional organization, as actioned by arkypallidal and prototypic neurons with specialized molecular, structural, and physiological properties, is fundamental to the operations of the dopamine-intact GPe. PMID:25926446

  4. An intact signal peptide on dengue virus E protein enhances immunogenicity for CD8(+) T cells and antibody when expressed from modified vaccinia Ankara.

    Science.gov (United States)

    Quinan, Bárbara R; Flesch, Inge E A; Pinho, Tânia M G; Coelho, Fabiana M; Tscharke, David C; da Fonseca, Flávio G

    2014-05-23

    Dengue is a global public health concern and this is aggravated by a lack of vaccines or antiviral therapies. Despite the well-known role of CD8(+) T cells in the immunopathogenesis of Dengue virus (DENV), only recent studies have highlighted the importance of this arm of the immune response in protection against the disease. Thus, the majority of DENV vaccine candidates are designed to achieve protective titers of neutralizing antibodies, with less regard for cellular responses. Here, we used a mouse model to investigate CD8(+) T cell and humoral responses to a set of potential DENV vaccines based on recombinant modified vaccinia virus Ankara (rMVA). To enable this study, we identified two CD8(+) T cell epitopes in the DENV-3 E protein in C57BL/6 mice. Using these we found that all the rMVA vaccines elicited DENV-specific CD8(+) T cells that were cytotoxic in vivo and polyfunctional in vitro. Moreover, vaccines expressing the E protein with an intact signal peptide sequence elicited more DENV-specific CD8(+) T cells than those expressing E proteins in the cytoplasm. Significantly, it was these same ER-targeted E protein vaccines that elicited antibody responses. Our results support the further development of rMVA vaccines expressing DENV E proteins and add to the tools available for dengue vaccine development.

  5. Study and ICH validation of a reverse-phase liquid chromatographic method for the quantification of the intact monoclonal antibody cetuximab

    Directory of Open Access Journals (Sweden)

    Antonio Martínez-Ortega

    2016-04-01

    Full Text Available Cetuximab (CTX is a potent chimeric mouse/human monoclonal antibody (mAb approved worldwide for treatment of metastatic colorectal cancer. Among the various biological and physical analyses performed for full study on this biopharmaceutic, the determination of the concentration preparations throughout manufacturing and subsequent handling in hospital is particularly relevant. In the present work, the study and validation of a method for quantifying intact CTX by reverse-phase high-performance liquid chromatography with diode array detection ((RPHPLC/DAD is presented. With that end, we checked the performance of a chromatographic method for quantifying CTX and conducted a study to validate the method as stability-indicating in accordance with the International Conference on Harmonization guidelines (ICH for biotechnological drugs; therefore, we evaluated linearity, accuracy, precision, detection and quantification limits, robustness and system suitability. The specificity of the method and the robustness of the mAb formulation against external stress factors were estimated by comprehensive chromatographic analysis by subjecting CTX to several informative stress conditions. As demonstrated, the method is rapid, accurate, and reproducible for CTX quantification. It was also successfully used to quantify CTX in a long-term stability study performed under hospital conditions.

  6. Study and ICH validation of a reverse-phase liquid chromatographic method for the quantification of the intact monoclonal antibody cetuximab$

    Institute of Scientific and Technical Information of China (English)

    Antonio Martínez-Ortega; Agustín Herrera; Antonio Salmerón-García; José Cabeza; Luis Cuadros-Rodríguez; Natalia Navas

    2016-01-01

    Cetuximab (CTX) is a potent chimeric mouse/human monoclonal antibody (mAb) approved worldwide for treatment of metastatic colorectal cancer. Among the various biological and physical analyses per-formed for full study on this biopharmaceutic, the determination of the concentration preparations throughout manufacturing and subsequent handling in hospital is particularly relevant. In the present work, the study and validation of a method for quantifying intact CTX by reverse-phase high-perfor-mance liquid chromatography with diode array detection ((RP)HPLC/DAD) is presented. With that end, we checked the performance of a chromatographic method for quantifying CTX and conducted a study to validate the method as stability-indicating in accordance with the International Conference on Harmo-nization guidelines (ICH) for biotechnological drugs; therefore, we evaluated linearity, accuracy, preci-sion, detection and quantification limits, robustness and system suitability. The specificity of the method and the robustness of the mAb formulation against external stress factors were estimated by compre-hensive chromatographic analysis by subjecting CTX to several informative stress conditions. As de-monstrated, the method is rapid, accurate, and reproducible for CTX quantification. It was also suc-cessfully used to quantify CTX in a long-term stability study performed under hospital conditions.

  7. Elemental and isotopic imaging to study biogeochemical functioning of intact soil micro-environments

    Science.gov (United States)

    Mueller, Carsten W.

    2017-04-01

    The complexity of soils extends from the ecosystem-scale to individual micro-aggregates, where nano-scale interactions between biota, organic matter (OM) and mineral particles are thought to control the long-term fate of soil carbon and nitrogen. It is known that such biogeochemical processes show disproportionally high reaction rates within nano- to micro-meter sized isolated zones ('hot spots') in comparison to surrounding areas. However, the majority of soil research is conducted on large bulk (> 1 g) samples, which are often significantly altered prior to analysis and analysed destructively. Thus it has previously been impossible to study elemental flows (e.g. C and N) between plants, microbes and soil in complex environments at the necessary spatial resolution within an intact soil system. By using nano-scale secondary ion mass spectrometry (NanoSIMS) in concert with other imaging techniques (e.g. scanning electron microscopy (SEM) and micro computed tomography (µCT)), classic analyses (isotopic and elemental analysis) and biochemical methods (e.g. GC-MS) it is possible to exhibit a more complete picture of soil processes at the micro-scale. I will present exemplarily results about the fate and distribution of organic C and N in complex micro-scale soil structures for a range of intact soil systems. Elemental imaging was used to study initial soil formation as an increase in the structural connectivity of micro-aggregates. Element distribution will be presented as a key to detect functional spatial patterns and biogeochemical hot spots in macro-aggregate functioning and development. In addition isotopic imaging will be demonstrated as a key to trace the fate of plant derived OM in the intact rhizosphere from the root to microbiota and mineral soil particles. Especially the use of stable isotope enrichment (e.g. 13CO2, 15NH4+) in conjunction with NanoSIMS allows to directly trace the fate of OM or nutrients in soils at the relevant scale (e.g. assimilate C

  8. Effect of cold-restraint stress and zinc acexamate on gastric mucus production in intact glands.

    Science.gov (United States)

    Escolar, G; Navarro, C; Sendros, S; Bulbena, O

    1987-11-01

    Gastric mucus content was morphometrically evaluated in gastric glands of normal and cold-restraint stressed rats. Variations induced by treatment with zinc acexamate (200 mg/kg p.o.) were also investigated. Stress decreased the glycoprotein content in glands located in areas of injury. However, in intact glands from the same animals, the glycoprotein content was increased and the proportion of sulphated macromolecules greatly augmented. Zinc acexamate reduced the severity of damage in stressed rats. Although it augmented mucus content it prevented the modification in sulphated macromolecules in these rats. These findings are discussed in relation to the role of gastric mucus in preventing gastric damage.

  9. Magnetic resonance imaging of intact globe superior subluxation into the intracranium

    Directory of Open Access Journals (Sweden)

    Nariman Nezami

    2012-01-01

    Full Text Available A 67-year-old man with right-sided blunt ocular trauma is reported here. Despite having received primary medical care, the patient complained of severe headache for 14 days. Initial computed tomography (CT indicated hematoma in the right frontal lobe. However, magnetic resonance imaging (MRI indicated that the right globe along with its optic nerve had been intactly dislocated into the intracranium and differentiated from hematoma. In this case, the significance of MRI, in blunt ocular trauma work-up, and also regaining successful ocular function are highlighted.

  10. Instant Effects of Radiofrequency Electromagnetic Wave on Hemoglobin in Single Living Intact Red Blood Cell

    Institute of Scientific and Technical Information of China (English)

    Cheng Can YAO; Xiao Kun LI; Yao Xiong HUANG

    2005-01-01

    The absorption spectrum of the hemoglobin (Hb) in single living intact red blood cell (RBC), exposed in 900 MHz radiofrequency electromagnetic wave (RF-EMW), was non-invasive,in situ, real-time measured by employing a highly sensitive fast multi-channel microspectrophotometer system. Both the absorption intensity and site of intracellular Hb were altered after RBCs were exposed in 900 MHz RF-EMW with power density at 5 mW/cm2. It was indicated that not only the concentration of Hb in living RBCs was decreased, but the molecular structure of Hb was changed by the RF-EMW action.

  11. Superresolution and Fluorescence Dynamics Evidence Reveal That Intact Liposomes Do Not Cross the Human Skin Barrier

    DEFF Research Database (Denmark)

    Dreier, Jes; Sørensen, Jens A; Brewer, Jonathan R

    2016-01-01

    In this study we use the combination of super resolution optical microscopy and raster image correlation spectroscopy (RICS) to study the mechanism of action of liposomes as transdermal drug delivery systems in human skin. Two different compositions of liposomes were applied to newly excised human...... skin, a POPC liposome and a more flexible liposome containing the surfactant sodium cholate. Stimulated emission depletion microscopy (STED) images of intact skin and cryo-sections of skin treated with labeled liposomes were recorded displaying an optical resolution low enough to resolve the 100 nm...

  12. APPLICATION OF INDEX TESTING METHODS IN DETERMINING MECHANICAL PROPERTIES OF INTACT ROCK MATERIAL

    Directory of Open Access Journals (Sweden)

    Ivana Dobrilović

    2010-12-01

    Full Text Available The review of index testing methods with detailed observations on the application of Schmidt rebound hardness and point load strength index in mechanical properties estimations of intact rock material is given in this paper. The results of conducted testing in determining applicability of the above-mentioned testing methods for the estimations of uniaxial compressive strength, elasticity modulus, tensile strength and flexural strength for three characteristic types of limestone from the roof layers of „Korenići“ deposit are also presented (the paper is published in Croatian.

  13. A Universal Protocol for Photochemical Covalent Immobilization of Intact Carbohydrates for the Preparation of Carbohydrate Microarrays

    Science.gov (United States)

    Wang, Huibin; Zhang, Yiming; Yuan, Xun; Chen, Yi; Yan, Mingdi

    2010-01-01

    A universal photochemical method has been established for the immobilization of intact carbohydrates and their analogues, and for the fabrication of carbohydrate microarrays. The method features the use of perfluorophenyl azide (PFPA)-modified substrates and the photochemical reaction of surface azido groups with printed carbohydrates. Various aldoses, ketoses, non-reducing sugars such as alditols and their derivatives can be directly arrayed on the PFPA-modified chips. The lectin-recognition ability of arrayed mannose, glucose and their oligo- and polysaccharides were confirmed using surface plasmon resonance imaging and laser-induced fluorescence imaging. PMID:21138274

  14. Hydrogen ions directly regulating the oligomerization state of Photosystem I in intact Spirulina platensis cells

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    H+ concentration induced-monomerization or trimerization of photosystem I (PSI) in cyanobacteria has never been directly observed. In this work, taking characteristic spectra for the trimers and monomers as the indicators, it was experimentally demonstrated that H+ could induce the oligomeric changes of PSI reaction centers in the intact Spirulina ,platensis cells and also in the isolated thylakoid membrane complexes. Especially, the higher concentration of H+ would induce the monomerization while the lower the trimerization, suggesting the electrostatic interaction should be mainly responsible forchanges in the oligomeric state of PSI in Spirulina platensis.

  15. Multiphoton imaging to distinguish grana and starch inside an intact leaf

    Science.gov (United States)

    Chen, Mei-Yu; Zhuo, Guan-Yu; Chen, Po-Fu; Wu, Pei-Chun; Liu, Tzu-Ming; Chu, Shi-Wei

    2013-02-01

    We have demonstrated a straightforward and noninvasive method to identify the distribution of grana and starch within an intact leaf. Grana and starch are the major functional structures for photosynthesis and energy storage of plant, respectively. Both exhibit highly ordered molecular structures and appear as micrometer-sized granules inside chloroplasts. In order to distinguish grana and starch, we used multiphoton microscopy, with simultaneous acquisition of two photon fluorescence (2PF) and second harmonic generation (SHG) signals. Consequently, SHG is found on both grana and starch while 2PF from chlorophyll indicates the identity of grana.

  16. Recycling of intact dense core vesicles in neurites of NGF-treated PC12 cells.

    Science.gov (United States)

    Bauer, Roslyn A; Khera, Rebecca S; Lieber, Janet L; Angleson, Joseph K

    2004-07-30

    Exocytic fusion in neuroendocrine cells does not always result in complete release of the peptide contents from dense core vesicles (DCVs). In this study, we use fluorescence imaging and immunoelectron microscopy to examine the retention, endocytosis and recycling of chromogranin B in DCVs of NGF-treated PC12 cells. Our results indicate that DCVs retained and retrieved an intact core that was available for subsequent exocytic release. The endocytic process was inhibited by cyclosporine A or by substitution of extracellular Ca(2+) with Ba(2+) and the total recycling time was less than 5 min.

  17. IODP Expeditions 309 and 312 Drill an Intact Section of Upper Oceanic Basement into Gabbros

    Directory of Open Access Journals (Sweden)

    Douglas S. Wilson

    2007-03-01

    Full Text Available The Integrated Ocean Drilling Program’s (IODP Expeditions 309 and 312 successfully completed the first sampling of an intact section of upper oceanic crust, through lavas and the sheeted dikes into the uppermost gabbros. Hole 1256D, which was initiated on the Ocean Drilling Program’s (ODP Leg 206, now penetrates to >1500 mbsf and >1250 m sub-basement. The first gabbroic rocks were encountered at 1407 mbsf. Below this, the hole penetrates ~100 m into a complex zone of fractionated gabbros intruded into contact metamorphosed dikes.

  18. The Fate of Dissolved Creosote Compounds in an Intact Fratured Clay Column

    DEFF Research Database (Denmark)

    Broholm, Kim; Arvin, Erik; Hansen, Asger

    1995-01-01

    The fate of 16 different organics typical for creosote was studied under aerobic conditions in a large intact fractured clay column experiment. Some of the organics (benzene, toluene, o-xylene, phenol, and o-cresol) were transported at the same rate as bromide through the fractured clay, whereas ...... estimated. This column experiment revealed that transport of organics through fractured clay is an important process. It may in many cases cause groundwater contamination of aquifers, sofar believed to be protected by overlying clay layers....

  19. Una técnica para la extirpación de los órganos vomeronasales en el armadillo Chaetophractus villosus. Abordaje desde la cavidad bucal - A surgical procedure for the ablation of the Vomeronasal Organs in the armadillo Chaetophractus villosus. Approach from oral cavity

    Directory of Open Access Journals (Sweden)

    Iodice, O. H.

    2010-03-01

    Full Text Available ResumenSe describen los pasos de un procedimiento quirúrgico para la extirpación bilateral de los órganos vomeronasales del armadillo Chaetophractus villosus (Mammalia, Xenarthra. Cada órgano está ubicado en el piso de la cavidad nasal, adosado a cada lado del tabique nasal. Los órganos se abordan a través de unaabertura practicada en el techo de la cavidad bucal. Este procedimiento se realiza bajo lupa estereoscópica y permite una visualización directa de los órganos Los mismos se destruyen por medio de una fresa accionada por un torno. De este modo se tiene la certeza de una ablación total. La posterior verificación por mediode cortes histológicos no es por lo tanto necesaria. Esto representa una ventaja importante puesto que las características físicas del hueso de estos animales requieren períodos largos de ablandamiento (generalmente alrededor tres meses. SummaryA surgical procedure for the bilateral ablation of the vomeronasal organs of the armadillo Chaetophractus villosus (Mammalia, Xenarthra is described. The organs are approached from the roof of the oral cavity through a longitudinal slit performed with a dental drill. This procedure is performed under magnifying stereoscopic lens and permits the direct visualization of the organs. Those structures are then destroyed with the dental drill. This provides certainty of totalablation. As a consequence, verification by means of histological sections is not needed. This represents an important advantage since the physical characteristics of the bone of these animals require long periods of softening (usually around three months.

  20. Cinacalcet reduces plasma intact parathyroid hormone, serum phosphate and calcium levels in patients with secondary hyperparathyroidism irrespective of its severity.

    LENUS (Irish Health Repository)

    2011-09-01

    To evaluate the relationship between the severity of secondary hyperparathyroidism (SHPT) - defined in terms of baseline plasma intact parathyroid hormone (iPTH) level - and the magnitude of response to cinacalcet.

  1. Novel molecular events associated with altered steroidogenesis induced by exposure to atrazine in the intact and castrate male rat

    Science.gov (United States)

    Toxicology is increasingly focused on molecular events comprising adverse outcome pathways. Atrazine activates the hypothalamic-pituitary adrenal axis, but relationships to gonadal alterations are unknown. We characterized hormone profiles and adrenal (intact and castrate) and te...

  2. Novel molecular events associated with altered steroidogenesis induced by exposure to atrazine in the intact and castrate male rat

    Science.gov (United States)

    Toxicology is increasingly focused on molecular events comprising adverse outcome pathways. Atrazine activates the hypothalamic-pituitary adrenal axis, but relationships to gonadal alterations are unknown. We characterized hormone profiles and adrenal (intact and castrate) and te...

  3. Lack of spatial segregation in the representation of pheromones and kairomones in the mouse medial amygdala.

    Directory of Open Access Journals (Sweden)

    Vinicius Miessler de Andrade Carvalho

    2015-08-01

    Full Text Available The nervous system is organized to detect, internally represent and process sensory information to generate appropriate behaviors. Despite the crucial importance of odors that elicit instinctive behaviors, such as pheromones and kairomones, their neural representation remains little characterized in the mammalian brain. Here we used expression of the immediate early gene product c-Fos as a marker of neuronal activity to find that a wide range of pheromones and kairomones produces activation in the medial nucleus of the amygdala, a brain area anatomically connected with the olfactory sensory organs. We see that activity in this nucleus depends on vomeronasal organ input, and that distinct vomeronasal stimuli activate a dispersed ensemble of cells, without any apparent spatial segregation. This activity pattern does not reflect the chemical category of the stimuli, their valence or the induced behaviors. These findings will help build a complete understanding of how odor information is processed in the brain to generate instinctive behaviors.

  4. Gravidez ovariana íntegra: tratamento cirúrgico videolaparoscópico The intact ovarian pregnancy: videolaparoscopy surgical treatament

    Directory of Open Access Journals (Sweden)

    Sergio Bigolin

    2007-04-01

    Full Text Available The intact ovarian pregnancy is an extremely rare condition. We report the case of a 34- years old woman case, nuliparus, with no previous clinical history of pelvic disease. An intact right ovarian pregnancy was diagnosed by a transvaginal ultrasonographic exam. A video-laparoscopy surgery was undertaken and only the ectopic tissue was removed. The authors discuss the effectiveness of the diagnostic tools and the surgical approach.

  5. Mouse pancreas tissue slice culture facilitates long-term studies of exocrine and endocrine cell physiology in situ.

    Directory of Open Access Journals (Sweden)

    Anja Marciniak

    Full Text Available Studies on pancreatic cell physiology rely on the investigation of exocrine and endocrine cells in vitro. Particularly, in the case of the exocrine tissue these studies have suffered from a reduced functional viability of acinar cells in culture. As a result not only investigations on dispersed acinar cells and isolated acini were limited in their potential, but also prolonged studies on pancreatic exocrine and endocrine cells in an intact pancreatic tissue environment were unfeasible. To overcome these limitations, we aimed to establish a pancreas tissue slice culture platform to allow long-term studies on exocrine and endocrine cells in the intact pancreatic environment. Mouse pancreas tissue slice morphology was assessed to determine optimal long-term culture settings for intact pancreatic tissue. Utilizing optimized culture conditions, cell specificity and function of exocrine acinar cells and endocrine beta cells were characterized over a culture period of 7 days. We found pancreas tissue slices cultured under optimized conditions to have intact tissue specific morphology for the entire culture period. Amylase positive intact acini were present at all time points of culture and acinar cells displayed a typical strong cell polarity. Amylase release from pancreas tissue slices decreased during culture, but maintained the characteristic bell-shaped dose-response curve to increasing caerulein concentrations and a ca. 4-fold maximal over basal release. Additionally, endocrine beta cell viability and function was well preserved until the end of the observation period. Our results show that the tissue slice culture platform provides unprecedented maintenance of pancreatic tissue specific morphology and function over a culture period for at least 4 days and in part even up to 1 week. This analytical advancement now allows mid -to long-term studies on the cell biology of pancreatic disorder pathogenesis and therapy in an intact surrounding in situ.

  6. Hot boning of intact carcasses: a procedure to avoid central nervous system self-contamination in beef and beef products.

    Science.gov (United States)

    Røtterud, Ole-Johan; Helps, Chris R; Hillman, Tim J; Fisher, Alan V; Harbour, Dave; Anil, Haluk; Nesbakken, Truls

    2006-02-01

    Compared with traditional boning of split refrigerated carcasses, hot boning of intact carcasses (the removal of meat from the skeleton prerigor) provides several commercially important cuts, may improve quality and reduce refrigeration costs, and may reduce the contamination of carcasses with central nervous system (CNS) tissue. In a comparative study of hot boning of intact and split carcasses, the CNS tissue contamination of intact carcasses was negligible (as measured with the CNS-related proteins glial fibrillary acidic protein and S-100 protein), but split carcasses were highly contaminated. The same trends were observed for dissection worktables used during the boning process. Most current boning plants have processing lines that are organized for boning carcass quarters, where the carcasses, in addition to transversal division, also are split horizontally. This part of the boning process was incorporated in the design of our study. Nine of the 18 intact carcasses were split horizontally between thoracic vertebrae 10 and 11 before they were hot boned. CNS tissue contamination was not detected on the carcass site related to this procedure. The amount of CNS tissue contamination was similar in boned cuts and minced meat from split and intact carcasses, except in the forerib. Boning of split carcasses appears to reduce CNS tissue contamination significantly to a level comparable to that of intact hot-boned carcasses.

  7. Visually guided whole cell patch clamp of mouse supraoptic nucleus neurons in cultured and acute conditions.

    Science.gov (United States)

    Stachniak, Tevye J E; Bourque, Charles W

    2006-07-01

    Recent advances in neuronal culturing techniques have supplied a new set of tools for studying neural tissue, providing effective means to study molecular aspects of regulatory elements in the supraoptic nucleus of the hypothalamus (SON). To combine molecular biology techniques with electrophysiological recording, we modified an organotypic culture protocol to permit transfection and whole cell patch-clamp recordings from SON cells. Neonatal mouse brain coronal sections containing the SON were dissected out, placed on a filter insert in culture medium, and incubated for at least 4 days to allow attachment to the insert. The SON was identifiable using gross anatomical landmarks, which remained intact throughout the culturing period. Immunohistochemical staining identified both vasopressinergic and oxytocinergic cells present in the cultures, typically appearing in well-defined clusters. Whole cell recordings from these cultures demonstrated that certain properties of the neonatal mouse SON were comparable to adult mouse magnocellular neurons. SON neurons in both neonatal cultures and acute adult slices showed similar sustained outward rectification above -60 mV and action potential broadening during evoked activity. Membrane potential, input resistance, and rapidly inactivating potassium current density (IA) were reduced in the cultures, whereas whole cell capacitance and spontaneous synaptic excitation were increased, perhaps reflecting developmental changes in cell physiology that warrant further study. The use of the outlined organotypic culturing procedures will allow the study of such electrophysiological properties of mouse SON using whole cell patch-clamp, in addition to various molecular, techniques that require longer incubation times.

  8. Structural organization of an intact phycobilisome and its association with photosystem II.

    Science.gov (United States)

    Chang, Leifu; Liu, Xianwei; Li, Yanbing; Liu, Cui-Cui; Yang, Fan; Zhao, Jindong; Sui, Sen-Fang

    2015-06-01

    Phycobilisomes (PBSs) are light-harvesting antennae that transfer energy to photosynthetic reaction centers in cyanobacteria and red algae. PBSs are supermolecular complexes composed of phycobiliproteins (PBPs) that bear chromophores for energy absorption and linker proteins. Although the structures of some individual components have been determined using crystallography, the three-dimensional structure of an entire PBS complex, which is critical for understanding the energy transfer mechanism, remains unknown. Here, we report the structures of an intact PBS and a PBS in complex with photosystem II (PSII) from Anabaena sp. strain PCC 7120 using single-particle electron microscopy in combination with biochemical and molecular analyses. In the PBS structure, all PBP trimers and the conserved linker protein domains were unambiguously located, and the global distribution of all chromophores was determined. We provide evidence that ApcE and ApcF are critical for the formation of a protrusion at the bottom of PBS, which plays an important role in mediating PBS interaction with PSII. Our results provide insights into the molecular architecture of an intact PBS at different assembly levels and provide the basis for understanding how the light energy absorbed by PBS is transferred to PSII.

  9. Patterns of electrical propagation in the intact pregnant guinea pig uterus.

    Science.gov (United States)

    Lammers, Wim J E P; Mirghani, H; Stephen, B; Dhanasekaran, S; Wahab, A; Al Sultan, M A H; Abazer, F

    2008-03-01

    Previous studies have reported on propagation of individual spikes in isolated segments of the pregnant uterus, but there is no information on patterns of spike propagation in the intact organ. There is also no information on propagation of myometrial burst. The aim of this study was to record, at high resolution, patterns of propagation of electrical activities in the pregnant uterus. Sixteen timed-pregnant guinea pigs were euthanized at term, and their uteruses isolated. Fetuses were removed and replaced by an equal amount of Tyrode. A 240-electrode array was positioned at various locations along the organ, all signals were recorded simultaneously, and the electrical propagations were reconstructed. In the intact pregnant uterus at term, spikes propagated with high velocity in longitudinal (6.8 +/- 2.4 cm/s) and slower velocity in circular direction (2.8 +/- 1.0 cm/s; P pregnant uterus reveals fundamental properties in spatial and temporal patterns of spike and burst propagation that determine the contraction of the organ.

  10. Displaced Medial and Lateral Bucket Handle Meniscal Tears With Intact ACL and PCL.

    Science.gov (United States)

    Boody, Barrett S; Omar, Imran M; Hill, James A

    2015-08-01

    Bucket handle lesions are vertical longitudinal tears in the meniscus that may displace centrally into the respective medial or lateral compartment, frequently causing mechanical symptoms, including pain, perceived instability, and mechanical locking. Bucket handle meniscal tears are most commonly from a traumatic etiology and are frequently found with concomitant anterior cruciate ligament (ACL) injuries. Multiple imaging signs and associations have been described for the diagnosis of bucket handle meniscus tears, including coronal truncation, absent bow tie sign, double posterior cruciate ligament (PCL), double ACL, displacement of the bucket handle fragment, and disproportionate posterior horn signs. Among meniscal pathology encountered on magnetic resonance imaging or during arthroscopy, bucket handle meniscal tears are infrequent occurrences. Furthermore, the occurrence of displaced medial and lateral bucket handle tears found on imaging and during arthroscopy is very uncommon and is only sparsely reported in the literature. When displaced medial and lateral bucket handle meniscal segments are visualized within the intercondylar notch along with the ACL and PCL, the radiologic findings are referred to as the "quadruple cruciate" sign or the "Jack and Jill lesion." Of the few case reports described in the literature, only one noted displaced medial and lateral bucket handle meniscus tears with an intact ACL and PCL. The current case report outlines a similar rare case of the quadruple cruciate sign: displaced medial and lateral bucket handle meniscal tears located within the intercondylar notch and an intact ACL and PCL.

  11. Dopant-specific unzipping of carbon nanotubes for intact crystalline graphene nanostructures

    Science.gov (United States)

    Lim, Joonwon; Narayan Maiti, Uday; Kim, Na-Young; Narayan, Rekha; Jun Lee, Won; Sung Choi, Dong; Oh, Youngtak; Min Lee, Ju; Yong Lee, Gil; Hun Kang, Seok; Kim, Hyunwoo; Kim, Yong-Hyun; Ouk Kim, Sang

    2016-01-01

    Atomic level engineering of graphene-based materials is in high demand to enable customize structures and properties for different applications. Unzipping of the graphene plane is a potential means to this end, but uncontrollable damage of the two-dimensional crystalline framework during harsh unzipping reaction has remained a key challenge. Here we present heteroatom dopant-specific unzipping of carbon nanotubes as a reliable and controllable route to customized intact crystalline graphene-based nanostructures. Substitutional pyridinic nitrogen dopant sites at carbon nanotubes can selectively initiate the unzipping of graphene side walls at a relatively low electrochemical potential (0.6 V). The resultant nanostructures consisting of unzipped graphene nanoribbons wrapping around carbon nanotube cores maintain the intact two-dimensional crystallinity with well-defined atomic configuration at the unzipped edges. Large surface area and robust electrical connectivity of the synergistic nanostructure demonstrate ultrahigh-power supercapacitor performance, which can serve for AC filtering with the record high rate capability of -85° of phase angle at 120 Hz.

  12. Human skin penetration of cobalt nanoparticles through intact and damaged skin.

    Science.gov (United States)

    Larese Filon, Francesca; Crosera, Matteo; Timeus, Elisa; Adami, Gianpiero; Bovenzi, Massimo; Ponti, Jessica; Maina, Giovanni

    2013-02-01

    Cobalt nanoparticles (CoNPs) are produced for several industrial and biomedical applications but there is a lack of data on human cutaneous absorption. Cobalt is also a skin sensitizer that can cause allergic contact dermatitis. Co applied as NPs, due to their small size and high surface, can penetrate into the skin in higher amount that bulk material. The aim of this study was to evaluate the absorption of Co applied as NPs in both intact and damaged skin. Experiments were performed using Franz cells and 1.0 mg cm(-2) of CoNPs was applied as donor phase for 24h. Mean Co content of 8.5 ± 1.2 ng cm(-2) and 1.87 ± 0.86 μg cm(-2) were found in the receiving solutions of Franz cells when the CoNPs suspension was applied on intact skin and on damaged skin, respectively. Twenty-four hours Co flux permeation was 76 ± 49 ng cm(-2)h(-1) in damaged skin with a lag time of 2.8 ± 2.1h. This study suggests that Co applied as NPs is able to penetrate the human skin in an in vitro diffusion cell system.

  13. Prediction of valid acidity in intact apples with Fourier transform near infrared spectroscopy

    Institute of Scientific and Technical Information of China (English)

    LIU Yan-de; YING Yi-bin; FU Xia-ping

    2005-01-01

    To develop nondestructive acidity prediction for intact Fuji apples, the potential of Fourier transform near infrared(FT-NIR) method with fiber optics in interactance mode was investigated. Interactance in the 800 nm to 2619 nm region was measured for intact apples, harvested from early to late maturity stages. Spectral data were analyzed by two multivariate calibration techniques including partial least squares (PLS) and principal component regression (PCR) methods. A total of 120 Fuji apples were tested and 80 of them were used to form a calibration data set. The influences of different data preprocessing and spectra treatments were also quantified. Calibration models based on smoothing spectra were slightly worse than that based on derivative spectra, and the best result was obtained when the segment length was 5 nm and the gap size was 10 points. Depending on data preprocessing and PLS method, the best prediction model yielded correlation coefficient of determination (r2) of 0.759, low root mean square error of prediction (RMSEP) of 0.0677, low root mean square error of calibration (RMSEC) of 0.0562. The results indicated the feasibility of FT-NIR spectral analysis for predicting apple valid acidity in a nondestructive way.

  14. IgE epitopes of intact and digested Ara h 1

    DEFF Research Database (Denmark)

    Bøgh, Katrine Lindholm; Nielsen, H.; Madsen, Charlotte Bernhard;

    2012-01-01

    epitopes have been suggested to be of great importance. ObjectiveThe aim of this study was to identify IgE specific epitopes of intact and digested Ara h 1, and to compare epitope patterns between humans and rats. MethodsSera from five peanut allergic patients and five Brown Norway rats were used...... to identify intact and digested Ara h 1-specific IgE epitopes by competitive immunoscreening of a phage-displayed random hepta-mer peptide library using polyclonal IgE from the individual sera. The resulting peptide sequences were mapped on the surface of a three-dimensional structure of the Ara h 1 molecule...... to mimic epitopes using a computer-based algorithm. ResultsPatients as well as rats were shown to have individual IgE epitope patterns. All epitope mimics were conformational and found to cluster into three different areas of the Ara h 1 molecule. Five epitope motifs were identified by patient IgE, which...

  15. Variable-angle total internal reflection fluorescence microscopy of intact cells of Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Kim Myung K

    2011-09-01

    Full Text Available Abstract Background Total internal reflection fluorescence microscopy (TIRFM is a powerful tool for observing fluorescently labeled molecules on the plasma membrane surface of animal cells. However, the utility of TIRFM in plant cell studies has been limited by the fact that plants have cell walls, thick peripheral layers surrounding the plasma membrane. Recently, a new technique known as variable-angle epifluorescence microscopy (VAEM was developed to circumvent this problem. However, the lack of a detailed analysis of the optical principles underlying VAEM has limited its applications in plant-cell biology. Results Here, we present theoretical and experimental evidence supporting the use of variable-angle TIRFM in observations of intact plant cells. We show that when total internal reflection occurs at the cell wall/cytosol interface with an appropriate angle of incidence, an evanescent wave field of constant depth is produced inside the cytosol. Results of experimental TIRFM observations of the dynamic behaviors of phototropin 1 (a membrane receptor protein and clathrin light chain (a vesicle coat protein support our theoretical analysis. Conclusions These findings demonstrate that variable-angle TIRFM is appropriate for quantitative live imaging of cells in intact tissues of Arabidopsis thaliana.

  16. Superresolution and Fluorescence Dynamics Evidence Reveal That Intact Liposomes Do Not Cross the Human Skin Barrier.

    Directory of Open Access Journals (Sweden)

    Jes Dreier

    Full Text Available In this study we use the combination of super resolution optical microscopy and raster image correlation spectroscopy (RICS to study the mechanism of action of liposomes as transdermal drug delivery systems in human skin. Two different compositions of liposomes were applied to newly excised human skin, a POPC liposome and a more flexible liposome containing the surfactant sodium cholate. Stimulated emission depletion microscopy (STED images of intact skin and cryo-sections of skin treated with labeled liposomes were recorded displaying an optical resolution low enough to resolve the 100 nm liposomes in the skin. The images revealed that virtually none of the liposomes remained intact beneath the skin surface. RICS two color cross correlation diffusion measurements of double labeled liposomes confirmed these observations. Our results suggest that the liposomes do not act as carriers that transport their cargo directly through the skin barrier, but mainly burst and fuse with the outer lipid layers of the stratum corneum. It was also found that the flexible liposomes showed a greater delivery of the fluorophore into the stratum corneum, indicating that they functioned as chemical permeability enhancers.

  17. Serum parathyroid hormone (PTH) in pregnant women determined by an immunoradiometric assay for intact PTH

    Energy Technology Data Exchange (ETDEWEB)

    Davis, O.K.; Hawkins, D.S.; Rubin, L.P.; Posillico, J.T.; Brown, E.M.; Schiff, I.

    1988-10-01

    Most studies of circulating PTH levels using traditional RIAs have supported the concept of physiological hyperparathyroidism of pregnancy, with pregnant women having serum immunoreactive PTH levels significantly higher than those in nonpregnant subjects. However, such RIAs are insensitive and often detect inactive PTH fragments, so that the correlation between PTH immunoreactivity and bioactivity is poor. Employing a new intact PTH immunoradiometric assay (Allegro-Nichols), we reassessed the effects of pregnancy on parathyroid function. The mean serum PTH level in 81 pregnant women was 14.4 +/- 6.3 (+/- SD) compared to 24.8 +/- 9.0 ng/L in 11 normally cycling nonpregnant women (P less than 0.001). The mean serum total and ionized calcium levels in the 2 groups were similar. In 5 of the pregnant women, serum bioactive PTH, determined by cytochemical bioassay, was slightly lower (7.7 +/- 3.4 ng/L) than in normal individuals (11.1 +/- 1.9 ng/L). Our findings suggest, in contrast with the results of most previous studies, that serum intact PTH may decline during pregnancy.

  18. Quantitative Determination of Fusarium proliferatum Concentration in Intact Garlic Cloves Using Near-Infrared Spectroscopy

    Science.gov (United States)

    Tamburini, Elena; Mamolini, Elisabetta; De Bastiani, Morena; Marchetti, Maria Gabriella

    2016-01-01

    Fusarium proliferatum is considered to be a pathogen of many economically important plants, including garlic. The objective of this research was to apply near-infrared spectroscopy (NIRS) to rapidly determine fungal concentration in intact garlic cloves, avoiding the laborious and time-consuming procedures of traditional assays. Preventive detection of infection before seeding is of great interest for farmers, because it could avoid serious losses of yield during harvesting and storage. Spectra were collected on 95 garlic cloves, divided in five classes of infection (from 1-healthy to 5-very highly infected) in the range of fungal concentration 0.34–7231.15 ppb. Calibration and cross validation models were developed with partial least squares regression (PLSR) on pretreated spectra (standard normal variate, SNV, and derivatives), providing good accuracy in prediction, with a coefficient of determination (R2) of 0.829 and 0.774, respectively, a standard error of calibration (SEC) of 615.17 ppb, and a standard error of cross validation (SECV) of 717.41 ppb. The calibration model was then used to predict fungal concentration in unknown samples, peeled and unpeeled. The results showed that NIRS could be used as a reliable tool to directly detect and quantify F. proliferatum infection in peeled intact garlic cloves, but the presence of the external peel strongly affected the prediction reliability. PMID:27428978

  19. In vitro comparison of human fibroblasts from intact and ruptured ACL for use in tissue engineering

    Directory of Open Access Journals (Sweden)

    T Brune

    2007-12-01

    Full Text Available The present study compares fibroblasts extracted from intact and ruptured human anterior cruciate ligaments (ACL for creation of a tissue engineered ACL-construct, made of porcine small intestinal submucosal extracellular matrix (SIS-ECM seeded with these ACL cells. The comparison is based on histological, immunohistochemical and RT-PCR analyses. Differences were observed between cells in a ruptured ACL (rACL and cells in an intact ACL (iACL, particularly with regard to the expression of integrin subunits and smooth muscle actin (SMA. Despite these differences in the cell source, both cell populations behaved similarly when seeded on an SIS-ECM scaffold, with similar cell morphology, connective tissue organization and composition, SMA and integrin expression. This study shows the usefulness of naturally occurring scaffolds such as SIS-ECM for the study of cell behaviour in vitro, and illustrates the possibility to use autologous cells extracted from ruptured ACL biopsies as a source for tissue engineered ACL constructs.

  20. Estimation of drug particle size in intact tablets by two dimensional X-ray diffractometry.

    Science.gov (United States)

    Thakral, Seema; Thakral, Naveen K; Suryanarayanan, Raj

    2017-09-09

    The average grain size of a crystalline material can be determined from the γ-profile of Debye rings in two-dimensional X-ray diffraction (2D XRD) frames. Our objectives were to: (i) validate the method for organic powders and use it to determine the grain size in intact tablets, and (ii) demonstrate the pharmaceutical application of this technique by determining the grain size of the active pharmaceutical ingredient (API) in marketed formulations. Six sieve fractions of sucrose were prepared and the particle size distribution was confirmed by laser diffraction. Their average grain size was determined from the 2D XRD frames by the γ-profile method. For particles size determined by the three methods were in good agreement. When these particles were compressed, there was no discernible change in the sucrose grain size in tablets. When the particles were > 250 μm, compression resulted in a mixture of large grains and fine powder. The grain size of acetaminophen in eleven marketed tablet formulations was determined to be either ∼ 35 μm or ∼ 80 μm. This non-destructive technique can therefore be potentially useful to estimate the grain size of crystalline formulation components in intact tablets. Copyright © 2017. Published by Elsevier Inc.

  1. In vitro bioactive properties of intact and enzymatically hydrolysed whey protein: targeting the enteroinsular axis.

    Science.gov (United States)

    Power-Grant, O; Bruen, C; Brennan, L; Giblin, L; Jakeman, P; FitzGerald, R J

    2015-03-01

    Enzymatically hydrolysed milk proteins have a variety of biofunctional effects some of which may be beneficial in the management of type 2 diabetes mellitus. The purpose of this study was to evaluate the effect of commercially available intact and hydrolysed whey protein ingredients (DH 32, DH 45) on markers of the enteroinsular axis (glucagon like peptide-1 secretion, dipeptidyl peptidase IV inhibition, insulin secretion and antioxidant activity) before and after simulated gastrointestinal digestion (SGID). A whey protein hydrolysate, DH32, significantly enhanced (P whey protein hydrolysates inhibited dipeptidyl peptidase IV activity, yielding half maximal inhibitory concentration values (IC50) of 1.5 ± 0.1 and 1.1 ± 0.1 mg mL(-1) for the DH 32 and DH 45, samples respectively, and were significantly more potent than the intact whey (P whey protein significantly enhanced (P whey, as measured by the oxygen radical absorbance capacity assay (ORAC). This antioxidant activity was maintained (DH 32, P > 0.05) or enhanced (DH 45, P whey stimulated GLP-1 secretion from enteroendocrine cells compared to vehicle control (P whey proteins and peptides can act through multiple targets within the enteroinsular axis and as such may have glucoregulatory potential.

  2. Evaluating a genetically encoded optical sensor of neural activity using electrophysiology in intact adult fruit flies

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    Gilles Laurent

    2007-11-01

    Full Text Available Genetically encoded optical indicators hold the promise of enabling non-invasive monitoring of activity in identified neurons in behaving organisms. However, the interpretation of images of brain activity produced using such sensors is not straightforward. Several recent studies of sensory coding used G-CaMP 1.3-a calcium sensor-as an indicator of neural activity; some of these studies characterized the imaged neurons as having narrow tuning curves, a conclusion not always supported by parallel electrophysiological studies. To better understand the possible cause of these conflicting results, we performed simultaneous in vivo 2-photon imaging and electrophysiological recording of G-CaMP 1.3 expressing neurons in the antennal lobe (AL of intact fruitflies. We find that G-CaMP has a relatively high threshold, that its signal often fails to capture spiking response kinetics, and that it can miss even high instantaneous rates of activity if those are not sustained. While G-CaMP can be misleading, it is clearly useful for the identification of promising neural targets: when electrical activity is well above the sensor's detection threshold, its signal is fairly well correlated with mean firing rate and G-CaMP does not appear to alter significantly the responses of neurons that express it. The methods we present should enable any genetically encoded sensor, activator, or silencer to be evaluated in an intact neural circuit in vivo in Drosophila.

  3. Gaze-cueing requires intact face processing - Insights from acquired prosopagnosia.

    Science.gov (United States)

    Burra, Nicolas; Kerzel, Dirk; Ramon, Meike

    2017-04-01

    Gaze-cueing is the automatic spatial orienting of attention in the direction of perceived gaze. Participants respond faster to targets located at positions congruent with the direction of gaze, compared to incongruent ones (gaze cueing effect, GCE). However, it still remains unclear whether its occurrence depends on intact integration of information from the entire eye region or face, rather than simply the presence of the eyes per se. To address this question, we investigated the GCE in PS, an extensively studied case of pure acquired prosopagnosia. In our gaze-cueing paradigm, we manipulated the duration at which cues were presented (70ms vs. 400ms) and the availability of facial information (full-face vs. eyes-only). For 70ms cue duration, we found a context-dependent dissociation between PS and controls: PS showed a GCE for eyes-only stimuli, whereas controls showed a GCE only for full-face stimuli. For 400ms cue duration, PS showed gaze-cueing independently of stimulus context, whereas in healthy controls a GCE again emerged only for full-face stimuli. Our findings suggest that attentional deployment based on the gaze direction of briefly presented faces requires intact processing of facial information, which affords salience to the eye region. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Proteomic analysis using 2-D liquid separations of intact proteins from whole-cell lysates.

    Science.gov (United States)

    Zhu, Kan; Yan, Fang; O'Neil, Kimberly A; Hamler, Rick; Lubman, David M; Lin, Linda; Barder, Timothy J

    2004-02-01

    This unit describes procedures for 2-D liquid separations of proteins from whole-cell lysates. Protocols for protein isoelectric point (pI) fractionation in the first dimension include the use of liquid isoelectric focusing (IEF) and chromatofocusing. The liquid IEF provides a pI-based fractionation using a batch-phase electrophoretic method, while chromatofocusing uses a column-based chromatographic method to generate the pH gradient. Using either method, a second-dimension fractionation is provided in the liquid phase using nonporous silica-based reversed-phase HPLC (NPS-RP-HPLC) to generate a 2-D liquid map of the protein content of the cell. The eluate of the 2-D liquid fractionation is directly coupled to a mass spectrometer for on-line detection of the intact molecular weights of proteins. As a result, a multidimensional map of protein expression is obtained that characterizes cellular proteins by pI, hydrophobicity, and intact molecular weight. Such expression maps are useful for differential proteomic comparison between different cell samples.

  5. Phenol increases intracellular [Ca2+] during twitch contractions in intact Xenopus skeletal myofibers.

    Science.gov (United States)

    Nogueira, Leonardo; Hogan, Michael C

    2010-11-01

    Phenol is a neurolytic agent used for management of spasticity in patients with either motoneuron lesions or stroke. In addition, compounds that enhance muscle contractility (i.e., polyphenols, etc.) may affect muscle function through the phenol group. However, the effects of phenol on muscle function are unknown, and it was, therefore, the purpose of the present investigation to examine the effects of phenol on tension development and Ca(2+) release in intact skeletal muscle fibers. Dissected intact muscle fibers from Xenopus laevis were electrically stimulated, and cytosolic Ca(2+) concentration ([Ca(2+)](c)) and tension development were recorded. During single twitches and unfused tetani, phenol significantly increased [Ca(2+)](c) and tension without affecting myofilament Ca(2+) sensitivity. To investigate the phenol effects on Ca(2+) channel/ryanodine receptors, single fibers were treated with different concentrations of caffeine in the presence and absence of phenol. Low concentrations of phenol significantly increased the caffeine sensitivity (P twitch contractions in muscle fibers without altering myofilament Ca(2+) sensitivity and may be used as a new agent to study skeletal muscle Ca(2+) handling.

  6. A genetically encoded tag for correlated light and electron microscopy of intact cells, tissues, and organisms.

    Directory of Open Access Journals (Sweden)

    Xiaokun Shu

    2011-04-01

    Full Text Available Electron microscopy (EM achieves the highest spatial resolution in protein localization, but specific protein EM labeling has lacked generally applicable genetically encoded tags for in situ visualization in cells and tissues. Here we introduce "miniSOG" (for mini Singlet Oxygen Generator, a fluorescent flavoprotein engineered from Arabidopsis phototropin 2. MiniSOG contains 106 amino acids, less than half the size of Green Fluorescent Protein. Illumination of miniSOG generates sufficient singlet oxygen to locally catalyze the polymerization of diaminobenzidine into an osmiophilic reaction product resolvable by EM. MiniSOG fusions to many well-characterized proteins localize correctly in mammalian cells, intact nematodes, and rodents, enabling correlated fluorescence and EM from large volumes of tissue after strong aldehyde fixation, without the need for exogenous ligands, probes, or destructive permeabilizing detergents. MiniSOG permits high quality ultrastructural preservation and 3-dimensional protein localization via electron tomography or serial section block face scanning electron microscopy. EM shows that miniSOG-tagged SynCAM1 is presynaptic in cultured cortical neurons, whereas miniSOG-tagged SynCAM2 is postsynaptic in culture and in intact mice. Thus SynCAM1 and SynCAM2 could be heterophilic partners. MiniSOG may do for EM what Green Fluorescent Protein did for fluorescence microscopy.

  7. Feasibility in multispectral imaging for predicting the content of bioactive compounds in intact tomato fruit.

    Science.gov (United States)

    Liu, Changhong; Liu, Wei; Chen, Wei; Yang, Jianbo; Zheng, Lei

    2015-04-15

    Tomato is an important health-stimulating fruit because of the antioxidant properties of its main bioactive compounds, dominantly lycopene and phenolic compounds. Nowadays, product differentiation in the fruit market requires an accurate evaluation of these value-added compounds. An experiment was conducted to simultaneously and non-destructively measure lycopene and phenolic compounds content in intact tomatoes using multispectral imaging combined with chemometric methods. Partial least squares (PLS), least squares-support vector machines (LS-SVM) and back propagation neural network (BPNN) were applied to develop quantitative models. Compared with PLS and LS-SVM, BPNN model considerably improved the performance with coefficient of determination in prediction (RP(2))=0.938 and 0.965, residual predictive deviation (RPD)=4.590 and 9.335 for lycopene and total phenolics content prediction, respectively. It is concluded that multispectral imaging is an attractive alternative to the standard methods for determination of bioactive compounds content in intact tomatoes, providing a useful platform for infield fruit sorting/grading.

  8. Modeling of Recovery Profiles in Mentally Disabled and Intact Patients after Sevoflurane Anesthesia; A Pharmacodynamic Analysis

    Science.gov (United States)

    Shin, Teo Jeon; Noh, Gyu-Jeong; Koo, Yong-Seo

    2014-01-01

    Purpose Mentally disabled patients show different recovery profiles compared to normal patients after general anesthesia. However, the relationship of dose-recovery profiles of mentally disabled patients has never been compared to that of normal patients. Materials and Methods Twenty patients (10 mentally disabled patients and 10 mentally intact patients) scheduled to dental surgery under general anesthesia was recruited. Sevoflurane was administered to maintain anesthesia during dental treatment. At the end of the surgery, sevoflurane was discontinued. End-tidal sevoflurane and recovery of consciousness (ROC) were recorded after sevoflurane discontinuation. The pharmacodynamic relation between the probability of ROC and end-tidal sevoflurane concentration was analyzed using NONMEM software (version VII). Results End-tidal sevoflurane concentration associated with 50% probability of ROC (C50) and γ value were lower in the mentally disabled patients (C50=0.37 vol %, γ=16.5 in mentally intact patients, C50=0.19 vol %, γ=4.58 in mentally disabled patients). Mentality was a significant covariate of C50 for ROC and γ value to pharmacodynamic model. Conclusion A sigmoid Emanx model explains the pharmacodynamic relationship between end-tidal sevoflurane concentration and ROC. Mentally disabled patients may recover slower from anesthesia at lower sevoflurane concentration at ROC an compared to normal patients. PMID:25323901

  9. On the utility of predictive chromatography to complement mass spectrometry based intact protein identification.

    Science.gov (United States)

    Pridatchenko, Marina L; Perlova, Tatyana Yu; Ben Hamidane, Hisham; Goloborodko, Anton A; Tarasova, Irina A; Gorshkov, Alexander V; Evreinov, Victor V; Tsybin, Yury O; Gorshkov, Mikhail V

    2012-03-01

    The amino acid sequence determines the individual protein three-dimensional structure and its functioning in an organism. Therefore, "reading" a protein sequence and determining its changes due to mutations or post-translational modifications is one of the objectives of proteomic experiments. The commonly utilized approach is gradient high-performance liquid chromatography (HPLC) in combination with tandem mass spectrometry. While serving as a way to simplify the protein mixture, the liquid chromatography may be an additional analytical tool providing complementary information about the protein structure. Previous attempts to develop "predictive" HPLC for large biomacromolecules were limited by empirically derived equations based purely on the adsorption mechanisms of the retention and applicable to relatively small polypeptide molecules. A mechanism of the large biomacromolecule retention in reversed-phase gradient HPLC was described recently in thermodynamics terms by the analytical model of liquid chromatography at critical conditions (BioLCCC). In this work, we applied the BioLCCC model to predict retention of the intact proteins as well as their large proteolytic peptides separated under different HPLC conditions. The specific aim of these proof-of-principle studies was to demonstrate the feasibility of using "predictive" HPLC as a complementary tool to support the analysis of identified intact proteins in top-down, middle-down, and/or targeted selected reaction monitoring (SRM)-based proteomic experiments.

  10. Intact error monitoring in combat Veterans with post-traumatic stress disorder.

    Science.gov (United States)

    Swick, Diane; Honzel, Nikki; Turken, U

    2015-11-30

    The error-related negativity (ERN) is a neuroelectric signature of performance monitoring during speeded response time tasks. Previous studies indicate that individuals with anxiety disorders show ERN enhancements that correlate with the degree of clinical symptomology. Less is known about the error monitoring system in post-traumatic stress disorder (PTSD). PTSD is characterized by impairments in the regulation of fear and other emotional responses, as well as deficits in maintaining cognitive control. Here, combat Veterans with PTSD were compared to control Veterans in two different versions of the flanker task (n=13 or 14 per group). Replicating and extending previous findings, PTSD patients showed an intact ERN in both experiments. In addition, task performance and error compensation behavior were intact. Finally, ERN amplitude showed no relationship with self-reported PTSD, depression, or post-concussive symptoms. These results suggest that error monitoring represents a relative strength in PTSD that can dissociate from cognitive control functions that are impaired, such as response inhibition and sustained attention. A healthy awareness of errors in external actions could be leveraged to improve interoceptive awareness of emotional state. The results could have positive implications for PTSD treatments that rely on self-monitoring abilities, such as neurofeedback and mindfulness training.

  11. Uranium migration through intact sandstone: effect of pollutant concentration and the reversibility of uptake

    Science.gov (United States)

    Sims, R.; Lawless, T. A.; Alexander, J. L.; Bennett, D. G.; Read, D.

    1996-02-01

    A series of core flood experiments has been performed to investigate the migration behaviour of uranium under rigidly controlled conditions. Intact sandstone cores, pre-equilibrated with synthetic groundwater, were flooded with uranium solutions at varying concentrations and the transport process monitored as a function of pH, tracer concentration and the concentration of a competing ion, cadmium. In each case a substantial amount of uranium was retained by the core, implying a strong interaction with the rock matrix. The adsorption process was found to be highly concentration dependent; however, such that the finite retention capacity of an intact core can be exhausted within a relatively short time. The reversibility of uptake was studied by attempting to displace adsorbed uranium by injected cadmium and flushing with brine. Most of the uranium was readily recoverable but a small percentage is released very slowly on flushing, suggesting conversion to a more stable form. Prior to performing each experiment a simulation was carried out using a one-dimensional coupled chemical transport code, encompassing a thermodynamic description of the electrical double layer. The model was successful in predicting a priori the dominant trends in the uranium migration behaviour which may aid in model developments for more complex geochemical regimes than those studied here.

  12. A laboratory experiment of intact polar lipid degradation in sandy sediments

    Directory of Open Access Journals (Sweden)

    J. Logemann

    2011-03-01

    Full Text Available Intact polar lipids (IPLs are considered biomarkers for living biomass. Their degradation in marine sediments, however, is poorly understood and complicates interpretation of their occurrence in geological samples. To investigate the turnover of IPLs, a degradation experiment with anoxic sandy sediments from the North Sea was conducted. Intact cells of two organisms that do not naturally occur in North Sea sediments were chosen as IPL sources: (i Saccharomyces cerevisiae, representative for ester-bound acyl lipids that also occur in Bacteria, and (ii the archaeon Haloferax volcanii, representative for ether-bound isoprenoid lipids. Surprisingly, IPLs with phosphoester-bound head groups showed approximately the same degradation rate as IPLs with glycosidic head groups. Furthermore, the results indicate a relatively fast degradation of S. cerevisiae IPLs with ester-bound moieties (analogs of bacterial membrane lipids and no significant degradation of archaeal IPLs with ether-bound moieties. Pore water and 16S rRNA-based DGGE analysis showed only a minor influence of the IPL source on microbial metabolism and community profiles. Due to our results, the IPL-based quantification of Archaea and Bacteria should be interpreted with caution.

  13. A laboratory experiment of intact polar lipid degradation in sandy sediments

    Directory of Open Access Journals (Sweden)

    J. Logemann

    2011-09-01

    Full Text Available Intact polar lipids (IPLs are considered biomarkers for living biomass. Their degradation in marine sediments, however, is poorly understood and complicates interpretation of their occurrence in geological samples. To investigate the turnover of IPLs, a degradation experiment with anoxic sandy sediments from the North Sea was conducted. Intact cells of two organisms that do not naturally occur in North Sea sediments were chosen as IPL sources: (i Saccharomyces cerevisiae, representative for ester-bound acyl lipids that also occur in Bacteria, and (ii the archaeon Haloferax volcanii, representative for ether-bound isoprenoid lipids. Surprisingly, IPLs with phosphoester-bound head groups showed approximately the same degradation rate as IPLs with glycosidic head groups. Furthermore, the results indicate a relatively fast degradation of S. cerevisiae IPLs with ester-bound moieties (analogs of bacterial membrane lipids and no significant degradation of archaeal IPLs with ether-bound moieties. Pore water and 16S rRNA-based DGGE analysis showed only a minor influence of the IPL source on microbial metabolism and community profiles. Due to our results, the IPL-based quantification of Archaea and Bacteria should be interpreted with caution.

  14. Mixed-mode reversed phase/positively charged repulsion chromatography for intact protein separation.

    Science.gov (United States)

    Ding, Ling; Guo, Zhimou; Hu, Zhuo; Liang, Xinmiao

    2017-05-10

    A mixed-mode reversed phase/positively charged repulsion stationary phase C8PN composed of octyl and amino group has been developed for separation of intact protein. Before the separation of proteins, a set of probe compounds were employed to evaluate the chromatographic properties of C8PN, demonstrating typical reversed phase/positively charged repulsion interaction on this stationary phase as estimated. Then the new C8PN stationary phase was used to separate a standard protein mixture on the reversed phase mode. Compared with a commercial C4 stationary phase, it showed different selectivity for some proteins. In order to better understand the properties of C8PN, the effect of acetonitrile content was investigated based on retention equation. Higher values of the equation parameters on C8PN demonstrated that the protein retentions were more sensitive to the change of acetonitrile content. Besides, the influences of buffer salt additives on the protein retentions were also studied. The retention factors of the proteins got larger with the increase of buffer salt concentration, which confirmed the positively charged repulsion interaction on the column. Finally, the C8PN was further applied to separate oxidized- and reduced- forms of Recombinant Human Growth Hormone. Our study indicated the advantages and application potential of mixed-mode reversed phase/positively charged repulsion stationary phase for intact protein separation. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Laboratory simulation of intact capture of cometary and asteroidal dust particles in ISAS

    Science.gov (United States)

    Fujiwara, A.; Nakamura, A.; Kadono, T.

    In order to develop a collector for intact capturing of cometary dust particles in the SOCCER mission and regolith dust particles released from asteroid surfaces by the impact of projectiles launched from a flying-by spacecraft, various kinds of materials as the collector candidates have been exposed to hypervelocity projectiles in our laboratory. Data based on the penetration characteristics of various materials (penetration depth, hole profile, effectiveness for intact capturing) are greatly increased. The materials tested for these simulation experiments include various kinds of low-density media and multisheet stacks; these are foamed plastics (polystyrene 0.01 g/cc), silica aerogels (0.04 g/cc), air (0.001 g/cc), liquid, and multisheet stack consisting of thin Al sheets (thickness 0.002 to 0.1 mm) or polyethylene sheets. Projectiles used are spheres or cylinders of nylon, polycarbonate, basalt, copper, iron, and volatile organics (e.g.,paradichlorobenzene) of size ranging from 30 micrometers to 1 cm launched by a two-stage light gas gun and a rail gun in ISAS at velocity up to about 7 km/s. Some results obtained by using nylon projectiles of velocity less than about 5 km/s are presented; the penetration depth vs. bulk density of the collector material for several kinds of materials and the velocity at which the projectiles begin to fragment vs. material density for foamed polystyrene.

  16. Sex Steroid Metabolism in Benign and Malignant Intact Prostate Biopsies: Individual Profiling of Prostate Intracrinology

    Directory of Open Access Journals (Sweden)

    Daniele Gianfrilli

    2014-01-01

    Full Text Available In vitro studies reveal that androgens, oestrogens, and their metabolites play a crucial role in prostate homeostasis. Most of the studies evaluated intraprostatic hormone metabolism using cell lines or preprocessed specimens. Using an ex vivo model of intact tissue cultures with preserved architecture, we characterized the enzymatic profile of biopsies from patients with benign prostatic hyperplasia (BPH or cancer (PC, focusing on 17β-hydroxy-steroid-dehydrogenases (17β-HSDs and aromatase activities. Samples from 26 men who underwent prostate needle core biopsies (BPH n = 14; PC n = 12 were incubated with radiolabeled 3H-testosterone or 3H-androstenedione. Conversion was evaluated by TLC separation and beta-scanning of extracted supernatants. We identified three major patterns of conversion. The majority of BPHs revealed no active testosterone/oestradiol conversion as opposed to prostate cancer. Conversion correlated with histology and PSA, but not circulating hormones. Highest Gleason scores had a higher androstenedion-to-testosterone conversion and expression of 17β-HSD-isoenzymes-3/5. Conclusions. We developed an easy tool to profile individual intraprostatic enzymatic activity by characterizing conversion pathways in an intact tissue environment. In fresh biopsies we found that 17β-HSD-isoenzymes and aromatase activities correlate with biological behaviour allowing for morphofunctional phenotyping of pathology specimens and clinical monitoring of novel enzyme-targeting drugs.

  17. [Management of threatening preterm labor with intact membranes: indications for antibiotics].

    Science.gov (United States)

    Winer, N

    2002-11-01

    To analyse the benefits and disadvantages of systematic antibiotic therapy in patient presenting a preterm labor with intact membranes. We reviewed French and English reports on Medline using to the following key words: "antibiotic therapy and preterm labor, preterm labor, streptococcus B, vaginose, mycoplasma, antenatal infection". The systematic prescription of antibiotics is not recommended for patients presenting preterm labor who have intact membranes and no symptoms of infection. The benefit of antibiotics is small and shows a tendency to prolong the pregnancy and the reduction of maternal infection. No benefit has been shown for neonatal results. When early-onset neonatal sepsis develops in a case in which antepartum chemoprophylaxis was used, the isolated bacteria will present an increasing risk of bacterial drug resistance. Local treatment (cream or pessary) do not belong in the treatment of threatening preterm labor and are not recommended for the prevention of prematurity or materno fetal infection. Risk groups of patients who present a positive vaginal colonization are subject to discussion. Studies do not allow us to ascertain that antibiotics have a beneficial effect on prematurity in these groups. Antibiotics are recommended for the treatment of asymptomatic bacteriuria. This treatment reduces prematurity and maternal infections. Despite poor consensus criteria, if threatening preterm labor is associated with a bacteriuria, experts usually recommend treatment.

  18. On plant detection of intact tomato fruits using image analysis and machine learning methods.

    Science.gov (United States)

    Yamamoto, Kyosuke; Guo, Wei; Yoshioka, Yosuke; Ninomiya, Seishi

    2014-07-09

    Fully automated yield estimation of intact fruits prior to harvesting provides various benefits to farmers. Until now, several studies have been conducted to estimate fruit yield using image-processing technologies. However, most of these techniques require thresholds for features such as color, shape and size. In addition, their performance strongly depends on the thresholds used, although optimal thresholds tend to vary with images. Furthermore, most of these techniques have attempted to detect only mature and immature fruits, although the number of young fruits is more important for the prediction of long-term fluctuations in yield. In this study, we aimed to develop a method to accurately detect individual intact tomato fruits including mature, immature and young fruits on a plant using a conventional RGB digital camera in conjunction with machine learning approaches. The developed method did not require an adjustment of threshold values for fruit detection from each image because image segmentation was conducted based on classification models generated in accordance with the color, shape, texture and size of the images. The results of fruit detection in the test images showed that the developed method achieved a recall of 0.80, while the precision was 0.88. The recall values of mature, immature and young fruits were 1.00, 0.80 and 0.78, respectively.

  19. On Plant Detection of Intact Tomato Fruits Using Image Analysis and Machine Learning Methods

    Directory of Open Access Journals (Sweden)

    Kyosuke Yamamoto

    2014-07-01

    Full Text Available Fully automated yield estimation of intact fruits prior to harvesting provides various benefits to farmers. Until now, several studies have been conducted to estimate fruit yield using image-processing technologies. However, most of these techniques require thresholds for features such as color, shape and size. In addition, their performance strongly depends on the thresholds used, although optimal thresholds tend to vary with images. Furthermore, most of these techniques have attempted to detect only mature and immature fruits, although the number of young fruits is more important for the prediction of long-term fluctuations in yield. In this study, we aimed to develop a method to accurately detect individual intact tomato fruits including mature, immature and young fruits on a plant using a conventional RGB digital camera in conjunction with machine learning approaches. The developed method did not require an adjustment of threshold values for fruit detection from each image because image segmentation was conducted based on classification models generated in accordance with the color, shape, texture and size of the images. The results of fruit detection in the test images showed that the developed method achieved a recall of 0.80, while the precision was 0.88. The recall values of mature, immature and young fruits were 1.00, 0.80 and 0.78, respectively.

  20. Monitoring intra- and extracellular redox capacity of intact barley aleurone layers responding to phytohormones.

    Science.gov (United States)

    Mark, Christina; Zór, Kinga; Heiskanen, Arto; Dufva, Martin; Emnéus, Jenny; Finnie, Christine

    2016-12-15

    Redox regulation is important for numerous processes in plant cells including abiotic stress, pathogen defence, tissue development, seed germination and programmed cell death. However, there are few methods allowing redox homeostasis to be addressed in whole plant cells, providing insight into the intact in vivo environment. An electrochemical redox assay that applies the menadione-ferricyanide double mediator is used to assess changes in the intracellular and extracellular redox environment in living aleurone layers of barley (Hordeum vulgare cv. Himalaya) grains, which respond to the phytohormones gibberellic acid and abscisic acid. Gibberellic acid is shown to elicit a mobilisation of electrons as detected by an increase in the reducing capacity of the aleurone layers. By taking advantage of the membrane-permeable menadione/menadiol redox pair to probe the membrane-impermeable ferricyanide/ferrocyanide redox pair, the mobilisation of electrons was dissected into an intracellular and an extracellular, plasma membrane-associated component. The intracellular and extracellular increases in reducing capacity were both suppressed when the aleurone layers were incubated with abscisic acid. By probing redox levels in intact plant tissue, the method provides a complementary approach to assays of reactive oxygen species and redox-related enzyme activities in tissue extracts.