Dorsal root ganglion neurons and tyrosine hydroxylase: An intriguing association with implications for sensation and pain

Tyrosine hydroxylase (TH) is a rate-limiting enzyme broadly expressed in noradrenergic and dopaminergic neurons in the central nervous system.57,70 Tyrosine hydroxylase is also expressed by peripheral sympathetic neurons98 and by enteric neurons within the gut.81,84 More than 30 years ago, TH was unexpectedly discovered in developing adult rodent cranial and dorsal root ganglion (DRG) neurons. Today, TH-expressing DRG neurons are being rediscovered as a relevant subpopulation. This review addresses the emerging importance of TH-expressing DRG neurons in sensation and pain mechanisms, focusing specifically on (1) their nature as C-low threshold mechanoreceptors (C-LTMRs); (2) their involvement in nociception/pain; and (3) their catecholaminergic phenotype.Fil: Brumovsky, Pablo Rodolfo. Universidad Austral. Facultad de Ciencias Biomédicas. Laboratorio de Investigaciones Biomédicas; Argentina. Universidad Austral. Facultad de Ciencias Biomédicas. Instituto de Investigaciones en Medicina Traslacional. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Medicina Traslacional; Argentin

First Experience with the Consolidation of WWER Reactor Pressure Vessel Knowledge through a New Method

One of today¿s activities of the Joint Research Centre¿s (JRC) Institute for Energy (IE) concerns data management and dissemination in nuclear safety. An ¿Online Data & Information Network¿ (ODIN) is set-up, which maintains one document database and four engineering databases. These databases aim to deploy networks for energy related research & development, specifically for nuclear energy and to provide the public experimental data of European projects on mechanical and thermo-physical material properties in comparison with international standards and recommendations. Due to its long lasting experience and being in a key position as regards web based d-base (e.g. ODIN), the IAEA for example has recently transferred the reactor surveillance data-base to the IE. Lately, many stakeholders, such as Institutes, R&D Organisations, Regulators, Utilities, Governmental Organisations, have recognised the need for collecting, preserving, consolidating (validating), and disseminating nuclear knowledge (documents, competences and data), in order to make it easily accessible to future generations through modern informatics tools and training and education measures. A broad spectrum of components and technologies should be considered, i.e. reactor pressure vessel (RPV), piping, internals, steam generator, etc. regarding knowledge, material data and practices. In the long run, it will also support future decommissioning exercises of nuclear installations as a valuable knowledge source. In addition to the knowledge in each Member State, the IE produced a long standing record of results from its own institutional activities and even more through the participation to a large number of European Network partnership projects. It is important, besides preservation, to consolidate the enormous amount of scientific results produced since. Therefore, the IE has developed a method for consolidation of nuclear knowledge. The method relays on the mobilisation of all identified leading experts in the EU in re-evaluating old knowledge and consolidating what is necessary to create training materials for the new generations. This method was applied for a pilot study for consolidating and preserving WWER RPV safety related knowledge, which is scattered in many countries and in different languages, facing a serious issue in terms of getting lost. This initiative could be the start of a wider Nuclear Knowledge Preservation and Consolidation activity. Experience gained from the first exercise will be presented in this paper.JRC.F.4-Safety of future nuclear reactor

Transcript expression of vesicular glutamate transporters in lumbar dorsal root ganglia and the spinal cord of mice – Effects of peripheral axotomy or hindpaw inflammation

Using specific riboprobes, we characterized the expression of vesicular glutamate transporter (VGLUT)1–VGLUT3 transcripts in lumbar 4–5 (L4–5) dorsal root ganglions (DRGs) and the thoracolumbar to lumbosacral spinal cord in male BALB/c mice after a 1- or 3-day hindpaw inflammation, or a 7-day sciatic nerve axotomy. Sham animals were also included. In sham and contralateral L4–5 DRGs of injured mice, VGLUT1-, VGLUT2- and VGLUT3 mRNAs were expressed in ∼45%, ∼69% or ∼17% of neuron profiles (NPs), respectively. VGLUT1 was expressed in large and medium-sized NPs, VGLUT2 in NPs of all sizes, and VGLUT3 in small and medium-sized NPs. In the spinal cord, VGLUT1 was restricted to a number of NPs at thoracolumbar and lumbar segments, in what appears to be the dorsal nucleus of Clarke, and in mid laminae III–IV. In contrast, VGLUT2 was present in numerous NPs at all analyzed spinal segments, except the lateral aspects of the ventral horns, especially at the lumbar enlargement, where it was virtually absent. VGLUT3 was detected in a discrete number of NPs in laminae III–IV of the dorsal horn. Axotomy resulted in a moderate decrease in the number of DRG NPs expressing VGLUT3, whereas VGLUT1 and VGLUT2 were unaffected. Likewise, the percentage of NPs expressing VGLUT transcripts remained unaltered after hindpaw inflammation, both in DRGs and the spinal cord. Altogether, these results confirm previous descriptions on VGLUTs expression in adult mice DRGs, with the exception of VGLUT1, whose protein expression was detected in a lower percentage of mouse DRG NPs. A detailed account on the location of neurons expressing VGLUTs transcripts in the adult mouse spinal cord is also presented. Finally, the lack of change in the number of neurons expressing VGLUT1 and VGLUT2 transcripts after axotomy, as compared to data on protein expression, suggests translational rather than transcriptional regulation of VGLUTs after injury.Fil: Malet, Mariana. Universidad Austral. Facultad de Ciencias Biomédicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Vieytes, C. A.. Universidad Austral. Facultad de Ciencias Biomédicas; ArgentinaFil: Lundgren, K. H.. University of Cincinnati; Estados UnidosFil: Seal, R. P.. University of Pittsburgh; Estados UnidosFil: Tomasella, María Eugenia. Universidad Austral. Facultad de Ciencias Biomédicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Seroogy, K. B.. University of Cincinnati; Estados UnidosFil: Hökfelt, T.. Karolinska Huddinge Hospital. Karolinska Institutet; SueciaFil: Gebhart, G. F.. University of Pittsburgh; Estados UnidosFil: Brumovsky, Pablo Rodolfo. Universidad Austral. Facultad de Ciencias Biomédicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. University of Pittsburgh; Estados Unido

Current status of the small punch test standardization within the ASTM

Since 2014, considerable attention has been paid to the standardization of small punch test technique within the American Society of Testing and Materials (ASTM). In 2016 large inter-laboratory study has been launched within the ASTM subcommittee E10.2 - Behavior and Use of Nuclear Structural Materials, involving 12 laboratories and 6 evaluated structural materials from the nuclear power plant components. Paper describes the current status of ASTM standardization, results of the inter-laboratory study, lessons learned and open questions remaining to be solved for the successful completion of the standardization process

Luminal hypertonicity and acidity modulate colorectal afferents and induce persistent visceral hypersensitivity

Carbohydrate malabsorption such as in lactose intolerance or enteric infection causes symptoms that include abdominal pain. Because this digestive disorder increases intracolonic osmolarity and acidity by accumulation of undigested carbohydrates and fermented products, we tested whether these two factors (hypertonicity and acidity) would modulate colorectal afferents in association with colorectal nociception and hypersensitivity. In mouse colorectum-pelvic nerve preparations in vitro, afferent activities were monitored after application of acidic hypertonic saline (AHS; pH 6.0, 800 mosM). In other experiments, AHS was instilled intracolonically to mice and behavioral responses to colorectal distension (CRD) measured. Application of AHS in vitro excited 80% of serosal and 42% of mechanically-insensitive colorectal afferents (MIAs), sensitizing a proportion of MIAs to become mechanically sensitive and reversibly inhibiting stretch-sensitive afferents. Acute intracolonic AHS significantly increased expression of the neuronal activation marker pERK in colon sensory neurons and augmented noxious CRD-induced behavioral responses. After three consecutive daily intracolonic AHS treatments, mice were hypersensitive to CRD 4-15 days after the first treatment. In complementary single fiber recordings in vitro, the proportion of serosal class afferents increased at day 4; the proportion of MIAs decreased, and muscular class stretch-sensitive afferents were sensitized at days 11-15 in mice receiving AHS. These results indicate that luminal hypertonicity and acidity, two outcomes of carbohydrate malabsorption, can induce colorectal hypersensitivity to distension by altering the excitability and relative proportions of colorectal afferents, suggesting the potential involvement of these factors in the development of abdominal pain.Fil: La, Jun Ho. University of Pittsburgh; Estados UnidosFil: Feng, Bin. University of Pittsburgh; Estados UnidosFil: Schwartz, Erica S.. University of Pittsburgh; Estados UnidosFil: Brumovsky, Pablo Rodolfo. Universidad Austral. Facultad de Ciencias Biomédicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gebhart, G. F.. University of Pittsburgh; Estados Unido

Use of agro-industrial wastes for the production of a wild yeast enzyme with disintegration activity on plant tissues

The objective of the present study was to develop a cost-effective medium, using agro-industrial wastes for the production of a polygalacturonase by Wickerhanomyces anomalus of interest in cassava starch industries. The effect of several raw agro-industrial wastes and others nutrients on polygalacturonase production by W. anomalus, were evaluated, in a reference fermentation medium, using statistical designs, by batch culture. The ability of the cell-free supernatant to extract cassava starch was evaluated. Lemon peel was the best inducer for the production of PGase. Statistical analysis of the data showed that lemon peel, Mg+2 and PO4HK2 had significant effect on PGase production, and the others variables (yeast extract, Ca+2, Fe+2, amino acid and trace element solution) were no significant. PGase synthesis reached ~31 EUmL-1, in the OFM (glucose, lemon peel, urea, vitamins, KH2PO4 and MgSO4), after 12 h of culture, at a lab scale bioreactor. PGase of W. anomalus, was able to disintegrate cassava tuber tissue, and the starch granules contained within the cells were released into the reaction medium. Lemon peel can be used as inducer for PGase production by W. anomalus, in a low cost culture medium, appropriate for the production of the enzyme at large scale.Fil: Maidana, Silvana Andrea. Universidad Nacional de Misiones. Facultad de Cs.exactas Químicas y Naturales. Departamento de Ciencia y Tecnol. de Los Alimentos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Esteche, Vanesa Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad Nacional de Misiones. Facultad de Cs.exactas Químicas y Naturales. Departamento de Ciencia y Tecnol. de Los Alimentos; ArgentinaFil: Hours, Roque Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; ArgentinaFil: Brumovsky, Luis Alberto. Universidad Nacional de Misiones. Facultad de Cs.exactas Químicas y Naturales. Departamento de Ciencia y Tecnol. de Los Alimentos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Martos, María Alicia. Universidad Nacional de Misiones. Facultad de Cs.exactas Químicas y Naturales. Departamento de Ciencia y Tecnol. de Los Alimentos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentin

Annealing and Re-embrittlement of Reactor Pressure Vessel Materials - State of the Art Report

Annealing of a reactor pressure vessel embrittled by neutron irradiation constitutes the only known technique to restore the initial material properties, to an extent that depends on the annealing conditions and on the materials. This technique is used in WWER-440 type reactor pressure vessels. A very important related issue is the one of re-embrittlement behaviour of the material after the annealing. In this respect, there is an obvious link with radiation embrittlement understanding. This report compiles the vast amount of information on annealing and re-embrittlement, which is available in the European countries where such annealing operations have been performed. In addition this topic was also investigated in various TACIS-PHARE projects, and the conclusions are included here as well. To complete the state-of-the-art, the results from a number of annealing experiments carried out in US on Western type RPV steels have also been considered.JRC.F.4-Nuclear design safet

Some lumbar sympathetic neurons develop a glutamatergic phenotype after peripheral axotomy with a note on VGLUT2-positive perineuronal baskets

Glutamate is the main excitatory neurotransmitter in the nervous system, including in primary afferent neurons. However, to date a glutamatergic phenotype of autonomic neurons has not been described. Therefore, we explored the expression of vesicular glutamate transporter (VGLUT) types 1, 2 and 3 in lumbar sympathetic chain (LSC) and major pelvic ganglion (MPG) of naïve BALB/C mice, as well as after pelvic nerve axotomy (PNA), using immunohistochemistry and in situ hybridization. Colocalization with activating transcription factor-3 (ATF-3), tyrosine hydroxylase (TH), vesicular acetylcholine transporter (VAChT) and calcitonin gene-related peptide was also examined. Sham-PNA, sciatic nerve axotomy (SNA) or naïve mice were included. In naïve mice, VGLUT2-like immunoreactivity (LI) was only detected in fibers and varicosities in LSC and MPG; no ATF-3-immunoreactive (IR) neurons were visible. In contrast, PNA induced upregulation of VGLUT2 protein and transcript, as well as of ATF-3-LI in subpopulations of LSC neurons. Interestingly, VGLUT2-IR LSC neurons coexpressed ATF-3, and often lacked the noradrenergic marker TH. SNA only increased VGLUT2 protein and transcript in scattered LSC neurons. Neither PNA nor SNA upregulated VGLUT2 in MPG neurons. We also found perineuronal baskets immunoreactive either for VGLUT2 or the acetylcholinergic marker VAChT in non-PNA MPGs, usually around TH-IR neurons. VGLUT1-LI was restricted to some varicosities in MPGs, was absent in LSCs, and remained largely unaffected by PNA or SNA. This was confirmed by the lack of expression of VGLUT1 or VGLUT3 mRNAs in LSCs, even after PNA or SNA. Taken together, axotomy of visceral and non-visceral nerves results in a glutamatergic phenotype of some LSC neurons. In addition, we show previously non-described MPG perineuronal glutamatergic baskets.Fil: Brumovsky, Pablo Rodolfo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Austral; Argentina. Univeristy of Pittsburgh. School of Medicine; Estados UnidosFil: Seroogy, Kim B.. University of Cincinnati; Estados UnidosFil: Lundgren, Kerstin H.. University of Cincinnati; Estados UnidosFil: Watanabe, Masahiko. Hokkaido University School of Medicine; JapónFil: Hökfelt, Tomas. Karolinska Huddinge Hospital. Karolinska Institutet; SueciaFil: Gebhart, G.F.. Univeristy of Pittsburgh. School of Medicine; Estados Unido

Expression of vesicular glutamate transporters in sensory and autonomic neurons innervating the mouse urinary bladder

Purpose: Vesicular glutamate transporters (VGLUTs), essential for loading glutamate into synaptic vesicles, are present in various neuronal systems. However, the expression of VGLUTs in neurons innervating the urinary bladder has not yet been analyzed. Here, we study the presence of VGLUTs type-1, -2 and -3 (VGLUT1, VGLUT2 and VGLUT3, respectively) in mouse urinary bladder neurons. Materials and Methods: Expression of VGLUT1, VGLUT2 and calcitonin gene-related peptide (CGRP) was analyzed by immunohistochemistry in retrogradely labeled primary afferent and autonomic neurons of BALB/C mice after injecting Fast Blue in the urinary bladder wall. To study VGLUT3, retrograde tracing of the urinary bladder was performed in transgenic mice where VGLUT3 is identified by detection of enhanced green fluorescent protein (EGFP). Results: Most urinary bladder DRG neurons expressed VGLUT2. A smaller percentage of neurons also expressed VGLUT1 or VGLUT3. Coexpression with CGRP was only observed for VGLUT2. Occasional VGLUT2-immunoreactive (IR) neurons were seen in the major pelvic ganglion (MPG). Abundant VGLUT2-IR nerves were detected in the urinary bladder dome, trigone and also the urethra; VGLUT1-IR nerves were discretely present. Conclusions: We present novel data on the expression of VGLUTs in sensory and autonomic neurons innervating the mouse urinary bladder. The frequent association of VGLUT2 and CGRP in sensory neurons suggests interactions between glutamatergic and peptidergic neurotransmissions, potentially influencing commonly perceived sensations in the urinary bladder, such as discomfort and pain.Fil: Brumovsky, Pablo Rodolfo. Universidad Austral. Facultad de Ciencias Biomédicas. Laboratorio de Investigaciones Biomédicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. University of Pittsburgh. Department of Anesthesiology. Pittsburgh Center for Pain Research; Estados UnidosFil: Seal, Rebecca P.. University of Pittsburgh. Department of Anesthesiology. Pittsburgh Center for Pain Research; Estados UnidosFil: Lundgren, Kerstin H.. University of Cincinnati. Department of Neurology; Estados UnidosFil: Seroogy, Kim B.. University of Cincinnati. Department of Neurology; Estados UnidosFil: Watanabe, Masahiko. Hokkaido University School of Medicine. Department of Anatomy; JapónFil: Gebhart, G. F.. University of Pittsburgh. Department of Anesthesiology. Pittsburgh Center for Pain Research; Estados Unido