Tomato Rab11a Characterization Evidenced a Difference Between SYP121-Dependent and SYP122-Dependent exocytosis

The regulatory functions of Rab proteins in membrane trafficking lie in their ability to perform as molecular switches that oscillate between a GTP- and a GDP-bound conformation. The role of tomato LeRab11a in secretion was analyzed in tobacco protoplasts. Green fluorescent protein (GFP)/red fluorescent protein (RFP)-tagged LeRab11a was localized at the trans-Golgi network (TGN) in vivo. Two serines in the GTP-binding site of the protein were mutagenized, giving rise to the three mutants Rab11S22N, Rab11S27N and Rab11S22/27N. The double mutation reduced secretion of a marker protein, secRGUS (secreted rat β-glucuronidase), by half, whereas each of the single mutations alone had a much smaller effect, showing that both serines have to be mutated to obtain a dominant negative effect on LeRab11a function. The dominant negative mutant was used to determine whether Rab11 is involved in the pathway(s) regulated by the plasma membrane syntaxins SYP121 and SYP122. Co-expression of either of these GFP-tagged syntaxins with the dominant negative Rab11S22/27N mutant led to the appearance of endosomes, but co-expression of GFP-tagged SYP122 also labeled the endoplasmic reticulum and dotted structures. However, co-expression of Rab11S22/27N with SYP121 dominant negative mutants decreased secretion of secRGUS further compared with the expression of Rab11S22/27N alone, whereas co-expression of Rab11S22/27N with SYP122 had no synergistic effect. With the same essay, the difference between SYP121- and SYP122-dependent secretion was then evidenced. The results suggest that Rab11 regulates anterograde transport from the TGN to the plasma membrane and strongly implicate SYP122, rather than SYP121. The differential effect of LeRab11a supports the possibility that SYP121 and SYP122 drive independent secretory event

Proposal of a Protocol for the Safe Removal of Post-Earthquake Provisional Shorings

The recent seismic events in Italy, including the earthquakes in L’Aquila in 2009 and central Italy in 2016, have significantly impacted the historical centers of small and medium-sized cities. These events directly affected their ancient masonry building heritage, resulting in severe damage. In order to minimize the risk of collapses and prevent further harm to people and structures until restoration efforts can be carried out, provisional post-seismic shorings have been extensively employed. These occurrences motivated several studies focused on the selection and assembly of post-seismic shorings, considering the various rigid failure mechanisms that may occur in a wall or section of an ancient masonry building. Yet, thus far, the critical considerations concerning the disassembly of these shorings, which significantly influence the repair process of a compromised structure from safety, organizational, and economic perspectives, have been overlooked. This research endeavors to establish a protocol for the dismantling of provisional shorings. To this end, a preliminary risk assessment tool has been devised, furnishing a safety index that correlates with the level of risk associated with shoring removal, along with corresponding risk categories. The study recommends preliminary interventions, categorized as mandatory or optional, to mitigate the risk prior to shoring removal. Furthermore, specific guidelines are provided based on the assessed risk level indicated by the safety index. To illustrate the application of this risk assessment tool, a case study involving an ancient masonry building in L’Aquila is presented

AtSYP51/52 functions diverge in the post-Golgi traffic and differently affect vacuolar sorting.

Plant SNAREs encoded by genes of the same subfamily are generally considered as redundant in promoting vesicle-associated membrane fusion events. Nonetheless the application of innovative experimental approaches highlighted that members of the same gene subfamily often have different functional specificities. In this work two closely related Qc-SNAREs, the AtSYP51 and the AtSYP52 are compared in their ability to influence different secretory pathways. Their role in the vesicle sorting to the central vacuole has been revised and they were found to have a novel inhibitory function. When transiently over-expressed, the SYP51 and the SYP52 distributed between the TGN and the tonoplast. Our data demonstrate that these SYPs act as t-SNARE when present on the membrane of TGN/PVC, whereas they behave as inhibitory or interfering SNAREs (i-SNAREs) when they accumulate on the tonoplast. Moreover, the performed functional analysis indicated that the AtSYP51 and the AtSYP52 role differ in the traffic to the vacuole. The findings are a novel contribution for the functional characterization of plant SNAREs that reveal additional non-fusogenic roles

Tomato Rab11a characterization evidenced a difference between SYP121 dependent and SYP122 dependent exocytosis

The regulatory functions of Rab proteins in membrane trafficking lie in their ability to perform as molecular switches that oscillate between a GTP- and a GDP-bound conformation. The role of tomato LeRab11a in secretion was analyzed in tobacco protoplasts. Green fluorescent protein (GFP)/red fluorescent protein (RFP)-tagged LeRab11a was localized at the trans-Golgi network (TGN) in vivo. Two serines in the GTP-binding site of the protein were mutagenized, giving rise to the three mutants Rab11S22N, Rab11S27N and Rab11S22/27N. The double mutation reduced secretion of a marker protein, secRGUS (secreted rat b-glucuronidase), by half, whereas each of the single mutations alone had a much smaller effect, showing that both serines have to be mutated to obtain a dominant negative effect on LeRab11a function. The dominant negative mutant was used to determine whether Rab11 is involved in the pathway(s) regulated by the plasma membrane syntaxins SYP121 and SYP122. Co-expression of either of these GFP-tagged syntaxins with the dominant negative Rab11S22/27N mutant led to the appearance of endosomes, but co-expression of GFP-tagged SYP122 also labeled the endoplasmic reticulum and dotted structures. However, co-expression of Rab11S22/27N with SYP121 dominant negative mutants decreased secretion of secRGUS further compared with the expression of Rab11S22/27N alone, whereas co-expression of Rab11S22/27N with SYP122 had no synergistic effect. With the same essay, the difference between SYP121- and SYP122-dependent secretion was then evidenced. The results suggest that Rab11 regulates anter- ograde transport from the TGN to the plasma membrane and strongly implicate SYP122, rather than SYP121. The differential effect of LeRab11a supports the possibility that SYP121 and SYP122 drive independent secretory events

Deletion of calcineurin from GFAP-expressing astrocytes impairs excitability of cerebellar and hippocampal neurons through astroglial Na+ /K+ ATPase

Astrocytes perform important housekeeping functions in the nervous system including maintenance of adequate neuronal excitability, although the regulatory mechanisms are currently poorly understood. The astrocytic Ca2+/calmodulin\u2010activated phosphatase calcineurin (CaN) is implicated in the development of reactive gliosis and neuroinflammation, but its roles, including the control of neuronal excitability, in healthy brain is unknown. We have generated a mouse line with conditional knockout (KO) of CaN B1 (CaNB1) in glial fibrillary acidic protein\u2010expressing astrocytes (astroglial calcineurin KO [ACN\u2010KO]). Here, we report that postnatal and astrocyte\u2010specific ablation of CaNB1 did not alter normal growth and development as well as adult neurogenesis. Yet, we found that specific deletion of astrocytic CaN selectively impairs intrinsic neuronal excitability in hippocampal CA1 pyramidal neurons and cerebellar granule cells (CGCs). This impairment was associated with a decrease in after hyperpolarization in CGC, while passive properties were unchanged, suggesting impairment of K+ homeostasis. Indeed, blockade of Na+/K+\u2010ATPase (NKA) with ouabain phenocopied the electrophysiological alterations observed in ACN\u2010KO CGCs. In addition, NKA activity was significantly lower in cerebellar and hippocampal lysates and in pure astrocytic cultures from ACN\u2010KO mice. While no changes were found in protein levels, NKA activity was inhibited by the specific CaN inhibitor FK506 in both cerebellar lysates and primary astroglia from control mice, suggesting that CaN directly modulates NKA activity and in this manner controls neuronal excitability. In summary, our data provide formal evidence for the notion that astroglia is fundamental for controlling basic neuronal functions and place CaN center\u2010stage as an astrocytic Ca2+\u2010sensitive switch

Interferon beta-1a in chronic inflammatory demyelinating polyneuropathy: case report Interferon beta en polineuropatía crónica inflamatoria desmienlinizante: caso clínico

Chronic inflammatory demyelinating polyneuropathy (CIDP) is an acquired immune-mediated neuropathy. It presents with a course of progression which may be slow and steady or step-wise or relapsing. Sensory ataxic polyneuropathy may be the only clinical manifestation of this disease. Treatment with interferon beta1a (INF beta1a) has been tried with different results in patients who were refractory to other, more conventional, immunomodulatory therapies. Here we report on a patient who had a relapsing form of pure sensory ataxic CIDP and who failed to respond to intravenous human immunoglobulin. He was put on INF beta1a for 3 years. During this period he suffered no relapses while his condition stabilized.<br>La polineuropatía crónica inflamatoria desmielinizante (PCID) es una neuropatía inmuno-mediada, que presenta un curso clínico primariamente progresivo o en forma de recaídas. Las manifestaciones sensoriales pueden ser su unica forma de expresión clínica. El tratamiento con interferon beta 1a (IFN beta1a) ha sido ensayado en varias oportunidades, con diferentes respuestas terapéuticas, en pacientes refractarios a las terapias inmunomoduladoras convencionales. Nosotros comunicamos un paciente con una forma ataxica recurrente de PCID, que no respondió al tratamiento con inmunoglobulina endovenosa. Posteriormente fue tratado con IFN beta 1 a por tres años. Durante el período de seguimiento no mostró nuevas recaídas y su cuadro neurológico se estabilizó

Multicentric epidemiological study in amyotrophic lateral sclerosis in the Autonomous City of Buenos Aires

Introduction: Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disease of unknown cause, characterized by the simultaneous involvement of the upper and lower motor neurons. Epidemiological studies have estimated its annual incidence between 0.31 and 3.2 and its prevalence between 0.8 and 8.5 cases per 100,000 inhabitants. The epidemiological information in our country is limited to specialized centers. The present study presents the results of an epidemiological study in ELA performed in the Autonomous City of Buenos Aires (CABA). Methods: A multicentric retrospective study was conducted. Patients with defined and probable ALS according to the El Escorial Criteria, evaluated between January 1, 2012 and December 31, 2013, who lived in the CABA at the onset of symptoms, were included. The calculation of the incidence was based on the 2010 census. Results: We included 103 patients (55 men), with a mean age of 64 years. The onset of symptoms was in the lower limbs at 39%, upper extremities at 25% and bulbar at 26%. The initial symptom was weakness in 58% and dysarthria in 20%; 9% had dementia associated with ALS. The mean time to diagnosis was 14.5 months. Thirty new cases/patients were diagnosed between 01/06/2012 and 01/06/2013, with an incidence rate of 1.04 per 100,000 inhabitants. Conclusions: The epidemiological characteristics of ALS in CABA are similar to those reported in the universal literature. Further studies are needed to determine if these findings are applicable to the rest of the Argentine population.Introduction: Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disease of unknown cause, characterized by the simultaneous involvement of the upper and lower motor neurons. Epidemiological studies have estimated its annual incidence between 0.31 and 3.2 and its prevalence between 0.8 and 8.5 cases per 100,000 inhabitants. The epidemiological information in our country is limited to specialized centers. The present study presents the results of an epidemiological study in ELA performed in the Autonomous City of Buenos Aires (CABA). Methods: A multicentric retrospective study was conducted. Patients with defined and probable ALS according to the El Escorial Criteria, evaluated between January 1, 2012 and December 31, 2013, who lived in the CABA at the onset of symptoms, were included. The calculation of the incidence was based on the 2010 census. Results: We included 103 patients (55 men), with a mean age of 64 years. The onset of symptoms was in the lower limbs at 39%, upper extremities at 25% and bulbar at 26%. The initial symptom was weakness in 58% and dysarthria in 20%; 9% had dementia associated with ALS. The mean time to diagnosis was 14.5 months. Thirty new cases/patients were diagnosed between 01/06/2012 and 01/06/2013, with an incidence rate of 1.04 per 100,000 inhabitants. Conclusions: The epidemiological characteristics of ALS in CABA are similar to those reported in the universal literature. Further studies are needed to determine if these findings are applicable to the rest of the Argentine population.Fil: Pérez Akly, Manuel. Sociedad Neurológica; ArgentinaFil: Schiava, Marianela. Unidad Asistencial Doctor César Milstein; ArgentinaFil: Melcom, Mario. Fundación para la Investigación en Neuroepidemiología; ArgentinaFil: Rodríguez, Gabriel. Sociedad Neurológica; ArgentinaFil: Gargiulo Monachelli, Gisella Mariana. Sociedad Neurológica; ArgentinaFil: Bettini, Mariela. Sociedad Neurológica; ArgentinaFil: Reisin, Ricardo. Sociedad Neurológica; ArgentinaFil: Bendersky, Mariana. Sociedad Neurológica; ArgentinaFil: Barroso, Fabio. Sociedad Neurológica; ArgentinaFil: Brand, Patricio. Fundación para la Lucha Contra las Enfermedades Neurológicas de la Infancia. Instituto de Investigaciones Neurológicas "Raúl Carrea"; ArgentinaFil: de Ambrosi, Bruno. Sociedad Neurológica; ArgentinaFil: Di Egidio, Marianna. Sociedad Neurológica; ArgentinaFil: Fiorotto, Luis. Sociedad Neurológica; ArgentinaFil: Jáuregui, Agustín. Sociedad Neurológica; ArgentinaFil: Landriscina, Paula. Sociedad Neurológica; ArgentinaFil: Marchesoni, Cintia. Sociedad Neurológica; ArgentinaFil: Mazia, Claudio. Sociedad Neurológica; ArgentinaFil: Rey, Roberto. Sociedad Neurológica; ArgentinaFil: Rugiero, Marcelo. Sociedad Neurológica; ArgentinaFil: Salutto, Valeria Luján. Sociedad Neurológica; ArgentinaFil: Tillard, Belén. Sociedad Neurológica; ArgentinaFil: Fulgenzi, Ernesto. Sociedad Neurológica; Argentin

Registry of neurological manifestations due to coronavirus-19 (COVID-19)

La enfermedad por COVID-19 se ha extendido por el mundo desde diciembre de 2019. Los síntomas neurológicos forman parte de su espectro clínico. Objetivo: Conocer las manifestaciones neurológicas en pacientes infectados por COVID-19 en Argentina. Métodos: Estudio multicéntrico realizado en adultos, desde mayo de 2020 a enero de 2021, con COVID-19 confirmado y síntomas neurológicos. Se consignaron variables demográficas, existencia de comorbilidades sistémicas o neurológicas, la forma de comienzo de la infección, alteración en estudios complementarios y el grado de severidad de los síntomas neurológicos. Resultados: Se incluyeron 817 pacientes de todo el país, 52% varones, edad promedio 38 anos. ˜ La mayoría sin comorbilidades ni patología neurológica previa. El primer síntoma de la infección fue neurológico en 56,2% de los casos, predominando la cefalea (69%), luego anosmia/ageusia (66%). También se reportaron mialgias (52%), alodinia/hiperalgesia (18%), astenia (6%). Un 3,2% mostró compromiso difuso del SNC como encefalopatía o convulsiones. Un 1,7% tuvo complicaciones cerebrovasculares. Los trastornos del sueno˜ se observaron en 3,2%. Se reportaron seis pacientes con síndrome de Guillain-Barré (GBS), neuropatía periférica (3,4%), parestesias en lengua (0,6%), hipoacusia (0,4%), plexopatía (0,3%). La severidad de síntomas neurológicos se correlacionó con la edad y la existencia de comorbilidades. Conclusiones: Nuestros resultados, similares a los de otros países, muestran dos tipos de síntomas neurológicos asociados a COVID-19: algunos potencialmente incapacitantes o mortales como el GBS o la encefalitis, y otros menos devastadores, pero más frecuentes, como cefalea o anosmia que demandan en forma creciente atención a largo plazo.COVID-19 disease has spread around the world since December 2019. Neurological symptoms are part of its clinical spectrum. Objective: To know the neurological manifestations in patients infected by COVID-19 in Argentina. Methods: Multicenter study conducted in adults, from May 2020 to January 2021, with confirmed COVID-19 and neurological symptoms. Demographic variables, existence of systemic or neurological comorbidities, the form of onset of the infection, alteration in complementary studies and the degree of severity of neurological symptoms were recorded. Results: 817 patients from all over the country were included, 52% male, mean age 38 years, most of them without comorbidities or previous neurological pathology. The first symptom of the infection was neurological in 56.2% of the cases, predominantly headache (69%), then anosmia/ageusia (66%). Myalgias (52%), allodynia/hyperalgesia (18%), and asthenia (6%) were also reported. 3.2% showed diffuse CNS involvement such as encephalopathy or seizures. 1.7% had cerebrovascular complications. Sleep disorders were observed in 3.2%. 6 patients were reported with Guillain Barré (GBS), peripheral neuropathy (3.4%), tongue paresthesia (0.6%), hearing loss (0.4%), plexopathy (0.3%). The severity of neurological symptoms was correlated with age and the existence of comorbidities. Conclusions: Our results, similar to those of other countries, show two types of neurological symptoms associated with COVID-19: some potentially disabling or fatal such as GBS or encephalitis, and others less devastating, but more frequent such as headache or anosmia that demand increasingly long-term care.Fil: Alessandro, Lucas. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; ArgentinaFil: Appiani, Franco. Fundación Favaloro; ArgentinaFil: Bendersky, Mariana. Universidad de Buenos Aires; ArgentinaFil: Borrego Guerrero, Brenda. Sanatorio Tandil; ArgentinaFil: Bruera, Guadalupe. Hospital Privado de Rosario; ArgentinaFil: Cairola, Patricia. Gobierno de la Ciudad Autónoma de Buenos Aires. Hospital General de Agudos Carlos Durand; ArgentinaFil: Calandri, Ismael. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; ArgentinaFil: Cardozo Oliver, Juan Martín. Sanatorio Finochietto; ArgentinaFil: Clément, María Emilia. Hospital Privado de la Comunidad; ArgentinaFil: Di Egidio, Marianna. Tornu Hospital; ArgentinaFil: Di Pace, José Luis. Gobierno de la Ciudad Autónoma de Buenos Aires. Hospital General de Agudos Carlos Durand; ArgentinaFil: Diaconchuk, Melina Alejandra. Hospital San Luis; ArgentinaFil: Esliman, Celeste. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; ArgentinaFil: Esnaola y Rojas, María Martha. Universidad de Buenos Aires; ArgentinaFil: Fernández Boccazzi, Julián. Sanatorio de la Trinidad; ArgentinaFil: Franco, Andrea Fabiana. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Ramos Mejía"; ArgentinaFil: Gargiulo Monachelli, Gisella Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; ArgentinaFil: Giardino, Daniela Laura. Centro de Educación Medica E Invest.clinicas; ArgentinaFil: Gómez, César. No especifíca;Fil: Guevara, Ana Karina. No especifíca;Fil: Gutiérrez, Natalia. Sanatorio Julio Méndez; ArgentinaFil: Hryb, Javier. Gobierno de la Ciudad Autónoma de Buenos Aires. Hospital General de Agudos Carlos Durand; ArgentinaFil: Ibarra, Viviana. Sanatorio Julio Méndez; ArgentinaFil: Janota, Franco. Universidad de Buenos Aires; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital de Infecciosas "Dr. Francisco Javier Muñiz"; ArgentinaFil: Larcher, Luis Alfredo. Sanatorio del Norte; ArgentinaFil: Leone, Fernando. Centro Médico Roca; ArgentinaFil: Luetic, Geraldine. No especifíca;Fil: Medina, Claudia Andrea. Sanatorio Las Lomas; ArgentinaFil: Menichini, María Laura. No especifíca;Fil: Nieto, Gonzalo. Hospital General de Agudos Bernardino Rivadavia ; Gobierno de la Ciudad Autonoma de Buenos Aires;Fil: Páez, María Fernanda. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Juan A. Fernández"; ArgentinaFil: Peñalver, Francisco. No especifíca;Fil: Perassolo, Mónica. Universidad de Buenos Aires; Argentina. Gobierno de la Ciudad Autónoma de Buenos Aires. Hospital General de Agudos Carlos Durand; ArgentinaFil: Persi, Gabriel. Universidad de Buenos Aires; ArgentinaFil: Pestchanker, Claudia. Hospital San Luis; ArgentinaFil: Porta, Oscar. Gobierno de la Ciudad Autónoma de Buenos Aires. Hospital General de Agudos Carlos Durand; ArgentinaFil: Rey, Roberto Daniel. Universidad de Buenos Aires; ArgentinaFil: Rodríguez, Gabriel Eduardo. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Ramos Mejía"; ArgentinaFil: Romano, Marina. Centro de Educación Medica E Invest.clinicas; ArgentinaFil: Rugiero, Marcelo. Hospital Italiano; ArgentinaFil: Saidón, Patricia. Universidad de Buenos Aires; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Ramos Mejía"; ArgentinaFil: Sica, María Florencia. Hospital Privado de la Comunidad; ArgentinaFil: Stankievich, Erica. No especifíca;Fil: Tarulla, Adriana. No especifíca;Fil: Zalazar, Guillermo. Hospital San Luis; Argentin