Los manantiales de agua salada de Rumanía explotados en tiempos arqueológicos : un nuevo candidato para el Patrimonio Mundial

Our approach emphasizes on the importance of the first forms of salt springs exploitation meant to obtain recrystallized salt for the development of prehistoric human communities within the continental inlands of Europe. Although it does not compare with the monumental dimension of World Heritage, the exploitation of some salt springs in Eastern Romania goes back around 8 millennia; they may be the oldest such exploitations in the world, as proven by 14C calibrated data. What differentiates Romanian salt springs from other famous similar areas in Europe is the continuity of exploitation and utilization of natural brine. Actually, these resilient behaviours explain the creation of a whole and complex universe of salt, which also represents a unique point of reference within the intangible World Heritage. It is through this association in variable proportions between tangible (non-monumental) and intangible that these salt springs comprising the oldest traces of salt exploitation can be considered elements of World Heritage. Today, important personalities in the fields of archaeology, anthropology and history posit that salt is a major reference for the development of the entire umanity. Obviously, the breakthrough of this idea requires awareness efforts targeting, on one hand, local communities in those areas wand, on the other, national and international scientific and cultural environments concerned with the World Heritage. In this context, a proper motivation is the fact that the last two decades have witnessed an intensification of research on salt, which turned this topic one of the major themes within European archaeology and ethno-archaeology. In terms of local community awareness concerning the importance of salt springs in the economic development of a (micro) area over time, it is worth underlining mostly the specialists’ efforts of presenting this topic in the media. Moreover, the impact of a recent initiative of the two museums in the area (Piatra Neamț and Târgu Neamț)—establishing distinct sections that represent, by using museum-inspired means, both archaeological vestiges and traditional practices of natural brine exploitation and utilization—will prove its extent in time. Certain local authorities and private entrepreneurs have pinpointed that valorising tourist areas comprising the oldest traces of salt exploitation in Romania is an imminent issue. The greatest challenge is finding a balance between the civilization improvements (upgraded access roads, upgrading operating areas, etc.) and thep rotection of still-alive traditional practices of salt exploitation and use, within rural areas. Certain local authorities and private entrepreneurs have pinpointed that valorising tourist areas comprising the oldest traces of salt exploitation in Romania will become, sooner or later, an imminent issue. The greatest challenge is finding a balance between the civilization improvements (upgraded access roads, upgrading operating areas, etc.) and the protection of still-alive traditional practices of salt exploitation and use, within rural areas.Nuestro planteamiento resalta la importancia de las primeras formas de explotación de manantiales de agua salada, que se emplearon en obtener sal recristalizada, para el desarrollo de las comunidades humanas prehistóricas ubicadas en la Europa continental. Aunque no puede compararse con la dimensión monumental del Patrimonio Mundial, la explotación de algunos manantiales de agua salada en el Este de Rumanía se remonta a unos 8 milenios atrás; podrían ser las más antiguas explotaciones de este tipo del mundo, según lo prueban las dataciones de C14 calibradas. Lo que diferencia a los manantiales de agua salada rumanos de otras zonas similares de Europa, reconocidas a nivel mundial, es la continuidad de las explotaciones y la utilización de la salmuera natural. De hecho, esta resiliencia comportamental explica la creación, en su conjunto, de un complejo universo de sal, que representa un punto de referencia único dentro del intangible Patrimonio Mundial. Mediante esta asociación, en proporciones variables, de lo intangible (nomonumental) y lo tangible, estos manantiales de agua salada que contienen los más antiguos rastros de la explotación de sal podrían ser considerados elementos del Patrimonio Mundial. Hoy en día, personalidades importantes del ámbito de la arqueología, antropología e historia postulan que la sal es una importante referencia para el desarrollo de toda la humanidad. Evidentemente, la imposición de esta idea requiere esfuerzos de concienciación, dirigidos hacia las comunidades locales, por un lado, y, por el otro, hacia el ámbito cultural y científico preocupado por el Patrimonio Mundial a nivel nacional e internacional. En este contexto, una motivación adecuada consiste en el hecho de que las últimas dos décadas han conocido una intensificación de las investigaciones acerca de la sal, que convirtieron este asunto en uno de los temas de mayor interés dentro de la arqueología y etnoarqueología europeas. En cuanto al grado de concienciación de las comunidades locales respecto a la importancia — a lo largo del tiempo — de los manantiales de agua salada para el desarrollo económico de un (micro) área, hay que resaltar los esfuerzos de los especialistas para conseguir cobertura mediática. Además, el impacto de una reciente propuesta de los dos museos del área (Piatra Neamț y Târgu Neamț) —de establecer secciones distintas que representen, empleando recursos museísticos, tanto vestigios arqueológicos como prácticas tradicionales de explotación y uso de la salmuera — quedará demostrado con el paso del tiempo. Algunas autoridades locales, junto con empresarios del ámbito privado señalaron que la valorización de zonas turísticas relacionadas con las huellas más antiguas de explotación de la sal en Rumanía es una cuestión de planteamiento inminente. El mayor reto es encontrar un equilibrio entre las mejoras impuestas por la civilización (caminos de acceso modernizados, áreas de funcionamiento perfeccionadas etc.) y la protección de las prácticas tradicionales de explotación y uso de la sal que aún perviven en las zonas rurales.Depto. de Prehistoria, Historia Antigua y ArqueologíaFac. de Geografía e HistoriaTRUEMinisterio de Economía y Competitividad (MINECO)pu

Repeated 5-day cycles of low dose aldesleukin in amyotrophic lateral sclerosis (IMODALS): a phase 2a randomised, double-blind, placebo-controlled trial

Background Low-dose interleukin-2 (ld-IL-2) enhances regulatory T-cell (Treg) function in auto-inflammatory conditions. Neuroinflammation being a pathogenic feature of amyotrophic lateral sclerosis (ALS), we evaluated the pharmacodynamics and safety of ld-IL-2 in ALS subjects. Methods We performed a single centre, parallel three-arm, randomised, double-blind, placebo-controlled study. Eligibility criteria included age 3 months, and a slow vital capacity ≥ 70% of normal. Patients were randomised (1:1:1) to aldesleukin 2 MIU, 1 MIU, or placebo once daily for 5 days every 4 weeks for 3 cycles. Primary outcome was change from baseline in Treg percentage of CD4+ T cells (%Tregs) following a first cycle. Secondary laboratory outcomes included: %Treg and Treg number following repeated cycles, and plasma CCL2 and neurofilament light chain protein (NFL) concentrations as surrogate markers of efficacy. Safety outcomes included motor-function (ALSFRS-R), slow vital capacity (SVC), and adverse event reports. This trial is registered with ClinicalTrials.gov, NCT02059759. Findings All randomised patients (12 per group), recruited from October 2015 to December 2015, were alive at the end of follow-up and included in the intent-to-treat (ITT) analysis. No drug-related serious adverse event was observed. Non-serious adverse events occurred more frequently with the 1 and 2 MIU IL-2 doses compared to placebo, including injection site reactions and flu-like symptoms. Primary outcome analysis showed a significant increase (p < 0·0001) in %Tregs in the 2 MIU and 1 MIU arms (mean [SD]: 2 MIU: +6·2% [2·2]; 1 MIU: +3·9% [1·2]) as compared to placebo (mean [SD]: -0·49% [1·3]). Effect sizes (ES) were large in treated groups: 2 MIU ES=3·7 (IC95%: 2·3–4·9) and 1 MIU ES=3·5 (IC95%: 2·1–4·6). Secondary outcomes showed a significant increase in %Tregs following repeated cycles (p < 0·0001) as compared to placebo, and a dose-dependent decrease in plasma CCL2 (p = 0·0049). There were no significant differences amongst the three groups on plasma NFL levels. Interpretation Ld-IL-2 is well tolerated and immunologically effective in subjects with ALS. These results warrant further investigation into their eventual therapeutic impact on slowing ALS disease progression

RNAseq Analyses Identify Tumor Necrosis Factor-Mediated Inflammation as a Major Abnormality in ALS Spinal Cord

ALS is a rapidly progressive, devastating neurodegenerative illness of adults that produces disabling weakness and spasticity arising from death of lower and upper motor neurons. No meaningful therapies exist to slow ALS progression, and molecular insights into pathogenesis and progression are sorely needed. In that context, we used high-depth, next generation RNA sequencing (RNAseq, Illumina) to define gene network abnormalities in RNA samples depleted of rRNA and isolated from cervical spinal cord sections of 7 ALS and 8 CTL samples. We aligned \u3e50 million 2X150 bp paired-end sequences/sample to the hg19 human genome and applied three different algorithms (Cuffdiff2, DEseq2, EdgeR) for identification of differentially expressed genes (DEG’s). Ingenuity Pathways Analysis (IPA) and Weighted Gene Co-expression Network Analysis (WGCNA) identified inflammatory processes as significantly elevated in our ALS samples, with tumor necrosis factor (TNF) found to be a major pathway regulator (IPA) and TNFα-induced protein 2 (TNFAIP2) as a major network “hub” gene (WGCNA). Using the oPOSSUM algorithm, we analyzed transcription factors (TF) controlling expression of the nine DEG/hub genes in the ALS samples and identified TF’s involved in inflammation (NFkB, REL, NFkB1) and macrophage function (NR1H2::RXRA heterodimer). Transient expression in human iPSC-derived motor neurons of TNFAIP2 (also a DEG identified by all three algorithms) reduced cell viability and induced caspase 3/7 activation. Using high-density RNAseq, multiple algorithms for DEG identification, and an unsupervised gene co-expression network approach, we identified significant elevation of inflammatory processes in ALS spinal cord with TNF as a major regulatory molecule. Overexpression of the DEG TNFAIP2 in human motor neurons, the population most vulnerable to die in ALS, increased cell death and caspase 3/7 activation. We propose that therapies targeted to reduce inflammatory TNFα signaling may be helpful in ALS patients

Human Umbilical Cord Blood Treatment in a Mouse Model of ALS: Optimization of Cell Dose

Amyotrophic Lateral Sclerosis (ALS) is a multicausal disease characterized by motor neuron degeneration in the spinal cord and brain. Cell therapy may be a promising new treatment for this devastating disorder. We recently showed that a single low dose (10(6) cells) of mononuclear human umbilical cord blood (MNC hUCB) cells administered intravenously to G93A mice delayed symptom progression and modestly prolonged lifespan. The aim of this pre-clinical translation study is to optimize the dose of MNC hUCB cells to retard disease progression in G93A mice. Three different doses of MNC hUCB cells, 10x10(6), 25x10(6) and 50x10(6), were administered intravenously into pre-symptomatic G93A mice. Motor function tests and various assays to determine cell effects were performed on these mice.Our results showed that a cell dose of 25x10(6) cells significantly increased lifespan of mice by 20-25% and delayed disease progression by 15%. The most beneficial effect on decreasing pro-inflammatory cytokines in the brain and spinal cord was found in this group of mice. Human Th2 cytokines were found in plasma of mice receiving 25x10(6) cells, although prevalent human Th1 cytokines were indicated in mice with 50x10(6) cells. High response of splenic cells to mitogen (PHA) was indicated in mice receiving 25x10(6) (mainly) and 10x10(6) cells. Significantly increased lymphocytes and decreased neutrophils in the peripheral blood were found only in animals receiving 25x10(6) cells. Stable reduction in microglia density in both cervical and lumbar spinal cords was also noted in mice administered with 25x10(6) cells.These results demonstrate that treatment for ALS with an appropriate dose of MNC hUCB cells may provide a neuroprotective effect for motor neurons through active involvement of these cells in modulating the host immune inflammatory system response

Skeletal Muscle Biopsy Analysis in Reducing Body Myopathy and Other Fhl1-related Disorders

FHL1 mutations have been associated with various disorders that include reducing body myopathy (RBM), Emery-Dreifuss-like muscular dystrophy, isolated hypertrophic cardiomyopathy, and some overlapping conditions. We report a detailed histochemical, immunohistochemical, electron microscopic, and immunoelectron microscopic analyses of muscle biopsies from 18 patients carrying mutations in FHL1: 14 RBM patients (Group 1), 3 Emery-Dreifuss muscular dystrophy patients (Group 2), and 1 patient with hypertrophic cardiomyopathy and muscular hypertrophy (Group 2). Group 1 muscle biopsies consistently showed RBs associated with cytoplasmic bodies. The RBs showed prominent FHL1 immunoreactivity whereas desmin, alpha B-crystallin, and myotilin immunoreactivity surrounded RBs. By electron microscopy, RBs were composed of electron-dense tubulofilamentous material that seemed to spread progressively between the myofibrils and around myonuclei. By immunoelectron microscopy, FHL1 protein was found exclusively inside RBs. Group 2 biopsies showed mild dystrophic abnormalities without RBs; only minor nonspecific myofibrillar abnormalities were observed under electron microscopy. Molecular analysis revealed missense mutations in the second FHL1 LIM domain in Group 1 patients and ins/del or missense mutations within the fourth FHL1 LIM domain in Group 2 patients. Our findings expand the morphologic features of RBM, clearly demonstrate the localization of FHL1 in RBs, and further illustrate major morphologic differences among different FHL1-related myopathies

Glycans in Sera of Amyotrophic Lateral Sclerosis Patients and Their Role in Killing Neuronal Cells

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease caused by degeneration of upper and lower motor neurons. To date, glycosylation patterns of glycoproteins in fluids of ALS patients have not been described. Moreover, the aberrant glycosylation related to the pathogenesis of other neurodegenerative diseases encouraged us to explore the glycome of ALS patient sera. We found high levels of sialylated glycans and low levels of core fucosylated glycans in serum-derived N-glycans of patients with ALS, compared to healthy volunteer sera. Based on these results, we analyzed the IgG Fc N297-glycans, as IgG are major serum glycoproteins affected by sialylation or core fucosylation and are found in the motor cortex of ALS patients. The analyses revealed a distinct glycan, A2BG2, in IgG derived from ALS patient sera (ALS-IgG). This glycan increases the affinity of IgG to CD16 on effector cells, consequently enhancing Antibody-Dependent Cellular Cytotoxicity (ADCC). Therefore, we explore whether the Fc-N297-glycans of IgG may be involved in ALS disease. Immunostaining of brain and spinal cord tissues revealed over-expression of CD16 and co-localization of intact ALS-IgG with CD16 and in brain with activated microglia of G93A-SOD1 mice. Intact ALS-IgG enhanced effector cell activation and ADCC reaction in comparison to sugar-depleted or control IgG. ALS-IgG were localized in the synapse between brain microglia and neurons of G93A-SOD1 mice, manifesting a promising in vivo ADCC reaction. Therefore, glycans of ALS-IgG may serve as a biomarker for the disease and may be involved in neuronal damage

Investigating the human—environment relationship of early intensive salt production: a case study from the Upper Seille Valley, Lorraine, northeast France

This paper presents the latest findings of multi-disciplinary research into the human—environment relationship of intensive Iron Age salt production in the Upper Seille Valley, Lorraine, northeast France. Investigations focus on the early Iron Age workshop “La Digue” (~ 625–500 cal BCE; Hallstatt D1–2), where high-resolution borehole sampling has been coupled with conventional excavation and geophysical surveying to establish direct linkages between intensive occupation and the alluvial environment of this site. Detailed insights into human—river interactions have been identified, enhancing current understanding of the environmental context and impact of this important early industry. The workshop's palaeogeographic setting has been reconstructed and new evidence for briquetage disposal practices has been identified, confirming that a close relationship existed between salt-making and the local hydrological regime. A large volume of briquetage waste (broken clay-fired salt-making equipment, ash and charcoal) was dumped into the river at La Digue, causing rapid and deliberate channel blockage, increasing the distance between the workshop and the river. This probably contributed to a localised increase in channel mobility and/or flooding whilst the workshop was active, producing challenging conditions for salt production. The workshop was abandoned following an intense flood event in ~ 500 cal BCE, coinciding with a major hydrological shift towards wetter floodplain conditions, likely arising from a combination of natural and anthropogenic factors. This study demonstrates the importance of understanding the environmental context of salt production and the roles of water management and briquetage disposal practices, which have been largely overlooked at other intensive salt making sites that employed the “briquetage technique”

Amyotrophic lateral sclerosis-motor neuron disease, monoclonal gammopathy, hyperparathyroidism, and B12 deficiency: case report and review of the literature

<p>Abstract</p> <p>Introduction</p> <p>Amyotrophic lateral sclerosis (the most common form of motor neuron disease) is a progressive and devastating disease involving both lower and upper motor neurons, typically following a relentless path towards death. Given the gravity of this diagnosis, all efforts must be made by the clinician to exclude alternative and more treatable entities. Frequent serology testing involves searching for treatable disorders, including vitamin B12 deficiency, parathyroid anomalies, and monoclonal gammopathies.</p> <p>Case presentation</p> <p>We present the case of a 78-year-old Caucasian man with all three of the aforementioned commonly searched for disorders during an investigation for amyotrophic lateral sclerosis.</p> <p>Conclusions</p> <p>The clinical utility of these common tests and what they ultimately mean in patients with amyotrophic lateral sclerosis is discussed, along with a review of the literature.</p

Evidence of Compromised Blood-Spinal Cord Barrier in Early and Late Symptomatic SOD1 Mice Modeling ALS

Background: The blood-brain barrier (BBB), blood-spinal cord barrier (BSCB), and blood-cerebrospinal fluid barrier (BCSFB) control cerebral/spinal cord homeostasis by selective transport of molecules and cells from the systemic compartment. In the spinal cord and brain of both ALS patients and animal models, infiltration of T-cell lymphocytes, monocyte-derived macrophages and dendritic cells, and IgG deposits have been observed that may have a critical role in motor neuron damage. Additionally, increased levels of albumin and IgG have been found in the cerebrospinal fluid in ALS patients. These findings suggest altered barrier permeability in ALS. Recently, we showed disruption of the BBB and BSCB in areas of motor neuron degeneration in the brain and spinal cord in G93A SOD1 mice modeling ALS at both early and late stages of disease using electron microscopy. Examination of capillary ultrastructure revealed endothelial cell degeneration, which, along with astrocyte alteration, compromised the BBB and BSCB. However, the effect of these alterations upon barrier function in ALS is still unclear. The aim of this study was to determine the functional competence of the BSCB in G93A mice at different stages of disease. Methodology/Principal Findings: Evans Blue (EB) dye was intravenously injected into ALS mice at early or late stage disease. Vascular leakage and the condition of basement membranes, endothelial cells, and astrocytes were investigated in cervical and lumbar spinal cords using immunohistochemistry. Results showed EB leakage in spinal cord microvessels from all G93A mice, indicating dysfunction in endothelia and basement membranes and confirming our previous ultrastructural findings on BSCB disruption. Additionally, downregulation of Glut-1 and CD146 expressions in the endothelial cells of the BSCB were found which may relate to vascular leakage. Conclusions/Significance: Results suggest that the BSCB is compromised in areas of motor neuron degeneration in ALS mice at both early and late stages of the disease