라틴아메리카 노동시장 및 노동정책의 현황과 전망

Carlos Salas,Christian Caldeira 한국노동연구원 2018 국제노동브리프 Vol.16 No.3

Riluzole Selective Antioxidant Effects in Cell Models Expressing Amyotrophic Lateral Sclerosis Endophenotypes

Gessica Sala,Alessandro Arosio,Elisa Conti,Simone Beretta,Christian Lunetta,Nilo Riva,Carlo Ferrarese,Lucio Tremolizzo 대한정신약물학회 2019 CLINICAL PSYCHOPHARMACOLOGY AND NEUROSCIENCE Vol.17 No.3

Objective: Until recently, riluzole was the only drug licensed for amyotrophic lateral sclerosis (ALS). In spite of its efficacy, the mechanism of action remains elusive, and both blocking of glutamate release and antioxidant properties have been postulated. Here we characterized human SH-SY5Y neuroblastoma cell lines, taking advantage of their insensitivity to excitotoxic insults, in order to selectively assess the presence of a direct antioxidant effect of riluzole. Methods: SH-SY5Y cells, either parental or overexpressing the G93A SOD1 mutation, were exposed for 24 hours to the selected stimuli. Results: Riluzole (1‒10 M) was able to counteract the effects of H2O2 exposure (200 M/24 hr), limiting both cell death and whole-cell reactive oxygen species (ROS) increase. The same experiments were repeated using SH-SY5Y cells carrying the familial ALS-related G93A-SOD1 mutation and constitutively expressing two-fold increased whole-cell ROS levels with respect to wild-type cells: riluzole was ineffective in this paradigm. Analogously, riluzole was ineffective in preventing cell death induced by exposing SH-SY5Y cells to 3-morpholino-sydnonimine (SIN-1, 1.5 mM/24 hr), a reactive nitrogen species (RNS) donor. Conclusion: Our data support a direct antioxidant action of riluzole. Furthermore, the lack of efficacy of riluzole observed in the SOD1 cell model mirrors the lack of efficacy already demonstrated in cognate mouse models of ALS, plausibly reflecting differences in the underlying pathogenic mechanisms. Finally, riluzole inefficacy against nitrosative stress might support the idea that a combined therapeutic intervention may result more effective in ALS patients, as in the case of co-administration of edaravone, a drug known to reduce RNS.

Sharpin, a novel postsynaptic density protein that directly interacts with the Shank family of proteins

Lim, Sangmi,Carlo Sala,Yoon, Jiyoung,Morgan Sheng,Kim, Eunjoon 이화여자대학교 세포신호전달연구센터 2000 고사리 세포신호전달 심포지움 Vol. No.2

The Shank family of proteins(also termed cortBP, ProSAP, or Synamon) are highly enriched in the postsynaptic density(PSD) of excitatory synapses in brain. Shank contains multiple domains for protein-protein interactions including ankyrin repeats, SH3 domain, PDZ domain, SAM domain and an extensive proline-rich region. Proteins that bind to Shank have been identified: the PDZ domain binds to the GKAP/SAPAP family of proteins, the proline-rich region interacts with Homer/Vesl and cortactin, and the SAM domain mediates homomultimerization of Shank. The functions of the N-terminal ankyrin repeats and the SH3 domain of Shank are unknown. We have identified a novel protein, termed Sharpin, that directly interacts with the N-terminal region of Shank containing the ankyrin repeats. Sharpin is widely expressed at mRNA and protein levels in many tissues and enriched in the PSD in brain. Sharpin forms a complex with Shank in heterologous cells and can be coimmunoprecipitated with Shank, GKAP and PSD-95 from brain extracts. Immunostaining reveals the presence of Sharpin at excitatory synapses and its partial colocalization with Shank. While the C-terminal half of Sharpin interacts with Shank, the N-terminal half of Sharpin mediates homomultimerization. These results define a new component of the postsynaptic PSD-95/GKAP/Shank complex at excitatory synapses.

Conservation and Valorization of Andean Crop Genetic Resources at the International Potato Center (CIP)

William M. Roca,Ana L. Panta,Ivan A. Manrique,Alberto R. Salas,Daniel M. Reynoso,Rene A. Gomez,Carlos Arbizu 한국원예학회 2006 한국원예학회 기타간행물 Vol.- No.-

Shear Rate and Microturbulence Effects on the Synthesis of Proteases by Jacaratia mexicana Cells Cultured in a Bubble Column, Airlift, and Stirred Tank Bioreactors

María del Carmen Oliver-Salvador,Elisa Morales-López,Enrique Durán-Páramo,Carlos Orozco-Álvarez,Sergio García-Salas 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.4

Cysteine proteases from Jacaratia mexicana,an endemic Mexican plant, could compete in industrial applications with papain. Currently the only way to obtain these proteases is by extracting them from the wild plant. An alternative source of these enzymes is by J. mexicana suspension culture. In this work, this culture was carried out in airlift, bubble column and stirred tank bioreactors,and the effects of shear rate and microturbulence on cell growth, protein accumulation and proteolytic activity were determined. The shear rates in the stirred tank, bubble column and airlift bioreactors were 274 1/s, 13 1/s and 36 1/s respectively, and microturbulences (symbolized by λ, in units of μm) were 46, 79, and 77 μm, respectively. Protein levels and proteolytic activity were linearly correlated with both shear rate and microturbulence. A higher shear rate and a more intensive microturbulence occurred in the stirred tank, producing higher protein accumulation and higher proteolytic activity compared with those of the other two bioreactor systems. Higher shear rate and microturbulence had an elicitor effect on protease synthesis, because microturbulence in stirred tank bioreactors was lower than the average length of J. mexicana cells. Furthermore, cells in the stirred tank were smaller and thinner than those grown in shake flask, bubble column and airlift bioreactors. In summary, proteases were produced by J. mexicana cell cultures in a stirred tank under conditions of high shear rate and intensive microturbulence, which are similar to those which occur in industrial stirred tanks. These results encourage continuation of the process development for large scale production of these proteases by this technology.

Metabolites in cultured cells of Ibervillea sonorae (S. Watson) Greene display increased hypoglycemic activity compared to that seen in plant roots

Ilse-Yazmín Arciniega-Carreón,María-Guadalupe Ramírez-Sotelo,Ana C. Ramos-Valdivia,Carlos Edmundo Salas,Arturo Ortega,Carmen Oliver-Salvador 한국원예학회 2020 Horticulture, Environment, and Biotechnology Vol.61 No.6

Ibervillea sonorae (S. Watson) Greene, (huereque) is an endemic shrub whose roots are used in traditional medicine to treatdiabetes. Here, we establish cell cultures of I. sonorae to evaluate the presence of antidiabetic metabolites. Stem and rootssections were incubated on media with diff erent concentrations of growth regulators to induce calli. Using B5 mediumcontaining 25 g L −1 sucrose, 150 mg L −1 ascorbic acid, 6 g L −1 agar–agar, supplemented with 0.5 mg L −1 α-naphthaleneaceticacid, 0.5 mg L −1 benzyladenine and 1 mg L −1 indole-3-acetic acid, 95% calli induction was attained. Flavonoids andphenols in I. sonorae cultured cell suspension were investigated. The methanol extracts exhibited antioxidant activity thatwas attributed to phenolic compounds. These extracts stimulated glucose uptake more effi caciously than I. sonorae plantroots. This study demonstrates that in vitro cultured cells stand as a good option for production of hypoglycemic metabolites.

“Over-inlay” block graft and differential morphometry: a novel block graft model to study bone regeneration and host-to-graft interfaces in rats

Giulia Ghiacci,Gallia Graiani,Francesca Ravanetti,Edoardo Manfredi,Carlo Galli,Antonio Cacchioli,Guido Maria Macaluso,Roberto Sala,Simone Lumetti 대한치주과학회 2016 Journal of Periodontal & Implant Science Vol.46 No.4

Purpose: The aim of this study was to present new a model that allows the study of the bone healing process, with an emphasis on the biological behavior of different graft-tohost interfaces. A standardized “over-inlay” surgical technique combined with a differential histomorphometric analysis is presented in order to optimize the use of critical-size calvarial defects in pre-clinical testing. Methods: Critical-size defects were created into the parietal bone of 8 male Wistar rats. Deproteinized bovine bone (DBBM) blocks were inserted into the defects, so that part of the block was included within the calvarial thickness and part exceeded the calvarial height (an “over-inlay” graft). All animals were sacrificed at 1 or 3 months. Histomorphometric and immunohistochemical evaluation was carried out within distinct regions of interest (ROIs): the areas adjacent to the native bone (BA), the periosteal area (PA) and the central area (CA). Results: The animals healed without complications. Differential morphometry allowed the examination of the tissue composition within distinct regions: the BA presented consistent amounts of new bone formation (NB), which increased over time (24.53%±1.26% at 1 month; 37.73%±0.39% at 3 months), thus suggesting that this area makes a substantial contribution toward NB. The PA was mainly composed of fibrous tissue (71.16%±8.06% and 78.30%±2.67%, respectively), while the CA showed high amounts of DBBM at both time points (78.30%±2.67% and 74.68%±1.07%, respectively), demonstrating a slow remodeling process. Blood vessels revealed a progressive migration from the interface with native bone toward the central area of the graft. Osterix-positive cells observed at 1 month within the PA suggested that the periosteum was a source of osteoprogenitor elements. Alkaline phosphatase data on matrix deposition confirmed this observation. Conclusions: The present model allowed for a standardized investigation of distinct graft-to-host interfaces both at vertically augmented and inlay-augmented sites, thus possibly limiting the number of animals required for pre-clinical investigations.

"Over-inlay" block graft and differential morphometry: a novel block graft model to study bone regeneration and host-to-graft interfaces in rats

Ghiacci, Giulia,Graiani, Gallia,Ravanetti, Francesca,Lumetti, Simone,Manfredi, Edoardo,Galli, Carlo,Cacchioli, Antonio,Macaluso, Guido Maria,Sala, Roberto Korean Academy of Periodontology 2016 Journal of Periodontal & Implant Science Vol.46 No.4

Purpose: The aim of this study was to present new a model that allows the study of the bone healing process, with an emphasis on the biological behavior of different graft-to-host interfaces. A standardized "over-inlay" surgical technique combined with a differential histomorphometric analysis is presented in order to optimize the use of critical-size calvarial defects in pre-clinical testing. Methods: Critical-size defects were created into the parietal bone of 8 male Wistar rats. Deproteinized bovine bone (DBBM) blocks were inserted into the defects, so that part of the block was included within the calvarial thickness and part exceeded the calvarial height (an "over-inlay" graft). All animals were sacrificed at 1 or 3 months. Histomorphometric and immunohistochemical evaluation was carried out within distinct regions of interest (ROIs): the areas adjacent to the native bone (BA), the periosteal area (PA) and the central area (CA). Results: The animals healed without complications. Differential morphometry allowed the examination of the tissue composition within distinct regions: the BA presented consistent amounts of new bone formation (NB), which increased over time ($24.53%{\pm}1.26%$ at 1 month; $37.73%{\pm}0.39%$ at 3 months), thus suggesting that this area makes a substantial contribution toward NB. The PA was mainly composed of fibrous tissue ($71.16%{\pm}8.06%$ and $78.30%{\pm}2.67%$, respectively), while the CA showed high amounts of DBBM at both time points ($78.30%{\pm}2.67%$ and $74.68%{\pm}1.07%$, respectively), demonstrating a slow remodeling process. Blood vessels revealed a progressive migration from the interface with native bone toward the central area of the graft. Osterix-positive cells observed at 1 month within the PA suggested that the periosteum was a source of osteoprogenitor elements. Alkaline phosphatase data on matrix deposition confirmed this observation. Conclusions: The present model allowed for a standardized investigation of distinct graft-to-host interfaces both at vertically augmented and inlay-augmented sites, thus possibly limiting the number of animals required for pre-clinical investigations.

Regulated RalBP1 Binding to RalA and PSD-95 Controls AMPA Receptor Endocytosis and LTD

Han, Kihoon,Kim, Myoung-Hwan,Seeburg, Daniel,Seo, Jinsoo,Verpelli, Chiara,Han, Seungnam,Chung, Hye Sun,Ko, Jaewon,Lee, Hyun Woo,Kim, Karam,Heo, Won Do,Meyer, Tobias,Kim, Hyun,Sala, Carlo,Choi, Se-Youn Public Library of Science 2009 PLoS biology Vol.7 No.9

<▼1><P>A two step mechanism was identified that regulates receptor endocytosis during the development of long-term depression (LTD), a long-lasting decrease in synaptic transmission.</P></▼1><▼2><P>Long-term depression (LTD) is a long-lasting activity-dependent decrease in synaptic strength. NMDA receptor (NMDAR)–dependent LTD, an extensively studied form of LTD, involves the endocytosis of AMPA receptors (AMPARs) via protein dephosphorylation, but the underlying mechanism has remained unclear. We show here that a regulated interaction of the endocytic adaptor RalBP1 with two synaptic proteins, the small GTPase RalA and the postsynaptic scaffolding protein PSD-95, controls NMDAR-dependent AMPAR endocytosis during LTD. NMDAR activation stimulates RalA, which binds and translocates widespread RalBP1 to synapses. In addition, NMDAR activation dephosphorylates RalBP1, promoting the interaction of RalBP1 with PSD-95. These two regulated interactions are required for NMDAR-dependent AMPAR endocytosis and LTD and are sufficient to induce AMPAR endocytosis in the absence of NMDAR activation. RalA in the basal state, however, maintains surface AMPARs. We propose that NMDAR activation brings RalBP1 close to PSD-95 to promote the interaction of RalBP1-associated endocytic proteins with PSD-95-associated AMPARs. This suggests that scaffolding proteins at specialized cellular junctions can switch their function from maintenance to endocytosis of interacting membrane proteins in a regulated manner.</P></▼2><▼3><P><B>Author Summary</B></P><P>Neurons adapt over time in order to dampen their response to prolonged or particularly strong stimuli. This process, termed long-term depression (LTD), results in a long-lasting decrease in the efficiency of synaptic transmission. One extensively studied form of LTD requires the activation of a specific class of receptors known as NMDA glutamate receptors (NMDARs). A key molecular event initiated by NMDA receptor activation is the stimulation of protein phosphatases. Another key event is internalization via endocytosis of synaptic AMPA glutamate receptors (AMPARs). However, the mechanism by which protein dephosphorylation is coupled to AMPAR endocytosis has remained unclear. Here, we help to define this mechanism. We show that endocytic proteins, including RalBP1, are widely distributed in neurons under normal conditions. Upon NMDAR activation, the small GTPase RalA becomes activated and binds to RalBP1, resulting in the translocation of RalBP1 and RalBP1-associated endocytic proteins to synapses. At the same time, RalBP1 becomes dephosphorylated, which promotes its binding to the postsynaptic scaffold protein PSD-95, a protein that itself associates with AMPARs. This concerted recruitment of endocytic proteins to the vicinity of AMPARs results in AMPAR endocytosis. On the basis of our data, we propose a model in which dual binding of RalBP1 to both RalA and PSD-95 following RalBP1 dephosphorylation is essential for NMDAR-dependent AMPAR endocytosis during LTD.</P></▼3>