Tracking Phenological Changes over 183 Years in Endemic Species of a Mediterranean Mountain (Sierra Nevada, SE Spain) Using Herbarium Specimens.

Phenological studies have a crucial role in the global change context. The Mediterranean basin constitutes a key study site since strong climate change impacts are expected, particularly in mountain areas such as Sierra Nevada, where we focus. Specifically, we delve into phenological changes in endemic vascular plants over time by analysing data at three scales: entire massif, altitudinal ranges, and particular species, seeking to contribute to stopping biodiversity loss. For this, we analysed 5262 samples of 2129 herbarium sheets from Sierra Nevada, dated from 1837 to 2019, including reproductive structure, complete collection date, and precise location. We found a generalized advancement in phenology at all scales, and particularly in flowering onset and flowering peak. Thus, plants flower on average 11 days earlier now than before the 1970s. Although similar trends have been confirmed for many territories and species, we address plants that have been studied little in the past regarding biotypes and distribution, and which are relevant for conservation. Thus, we analysed phenological changes in endemic plants, mostly threatened, from a crucial hotspot within the Mediterranean hotspot, which is particularly vulnerable to global warming. Our results highlight the urgency of phenological studies by species and of including ecological interactions and effects on their life cycles.

Soil filtration-sedimentation improves shelled protist recovery in eukaryotic eDNA surveys.

A large part of the soil protist diversity is missed in metabarcoding studies based on 0.25 g of soil environmental DNA (eDNA) and universal primers due to ca. 80% co-amplification of non-target plants, animals and fungi. To overcome this problem, enrichment of the substrate used for eDNA extraction is an easily implemented option but its effect has not yet been tested. In this study, we evaluated the effect of a 150 µm mesh size filtration and sedimentation method to improve the recovery of protist eDNA, while reducing the co-extraction of plant, animal and fungal eDNA, using a set of contrasted forest and alpine soils from La Réunion, Japan, Spain and Switzerland. Total eukaryotic diversity was estimated by V4 18S rRNA metabarcoding and classical amplicon sequence variant calling. A 2- to 3-fold enrichment in shelled protists (Euglyphida, Arcellinida and Chrysophyceae) was observed at the sample level with the proposed method, with, at the same time, a 2-fold depletion of Fungi and a 3-fold depletion of Embryophyceae. Protist alpha diversity was slightly lower in filtered samples due to reduced coverage in Variosea and Sarcomonadea, but significant differences were observed in only one region. Beta diversity varied mostly between regions and habitats, which explained the same proportion of variance in bulk soil and filtered samples. The increased resolution in soil protist diversity estimates provided by the filtration-sedimentation method is a strong argument in favour of including it in the standard protocol for soil protist eDNA metabarcoding studies.

Trade-Off between Facilitation and Interference of Allelopathic Compounds in Vegetation Recovery: The Case of Rosmarinus officinalis in Degraded Gypsum Habitats.

Rosmarinus officinalis advantageously competes with other species in restored gypsum outcrops, and further research is needed to understand the causes. Specifically, we focus on the potential allelopathic effects derived from its terpenes on the emergence of gypsum species. To this end, we established 120 circular subplots in a previously restored gypsum outcrop, and randomly applied four different treatments based on the presence/absence of rosemary plants and their leaves on the soil. Afterwards, we conducted an experimental sowing of native gypsophiles. All subplots were monitored to estimate seedling emergence, and soil and leaf samples were analysed for terpenes. The results show that the treatments had significant effects on the overall emergence of seedlings, and terpenes were found in rosemary leaves and soils, with no significant differences in terpene composition. In particular, we identified a clear negative effect in the treatment where rosemary plants were eliminated but its leaves were left along with allelopathy (2.57 ± 0.54 individuals/subplot). Unexpectedly, the presence of rosemary plants seems to facilitate the emergence of gypsum species (9.93 ± 1.61 individuals/subplot), counteracting the effects of the allelopathic substances in the soil. Consequently, we do not suggest removing rosemary plants in early stages to encourage the emergence of gypsum species in restored areas.

Assessing effectiveness of exclusion fences in protecting threatened plants.

Overgrazing stands out as threat factors on biodiversity, being especially harmful in the Mediterranean, due to strong human pressure and an accelerated climate change acting synergistically. Fencing is a common tool used in conservation biology to tackle this problem. Advantages of fences are usually fast, intuitive, and easy to evaluate. However, disadvantages could also arise (increasing interspecific competition, disturbing habitat structure, limiting pollination, reducing dispersion). Together with management issues (maintenance, conflicts with stakeholders, and pulling effect). Effectiveness of fencing for conservation has been frequently assessed for animals, while it is almost a neglected topic in plants. We evaluated the outcome of fencing three threatened and narrow-endemic plants. Selected 5 populations were only partly fenced, which allowed comparing different variables inside and outside the fence. For evaluating the fencing effects, we sampled several habitats (vegetation cover, composition, density of target species), and target-species features (individual size, neighbouring species, and fruit-set). Fencing had strong effects on the habitat and on target-species individuals, showing contrasting responses at species and population level. Particularly, for Erodium cazorlanum, fence had a positive effect in one case, and negative in another. In Hormathophylla baetica effect was positive in all populations. Finally, fencing negatively affected Solenanthus reverchonii by increasing competition and limiting seed-dispersal. Fencing outcome was different in assessed species, highlighting the need to a case-by-case evaluation to determine the net balance (pros vs. cons), also its suitability and most favourable option (i.e. permanent vs. temporary fences).

FloraSNevada: a trait database of the vascular flora of Sierra Nevada, southeast Spain.

Providing a complete data set with species and trait information for a given area is essential for assessing plant conservation, management, and ecological restoration, for both local and global applications. Also, these data sets provide additional information for surveys or data collections, establishing the starting point for more detailed studies on plant evolution, vegetation dynamics, and vegetation responses to disturbance and management. This data base covers Sierra Nevada mountains (southeastern Spain), a recognized plant biodiversity hotspot within the Mediterranean context. According to previous available data (before this augmented compilation), these mountains host 7% of the 24,000 Mediterranean vascular plants, despite covering just 0.01% of its area. Another characteristic of the Sierra Nevada is the great singularity of its flora, with 95 taxa being endemic to the high-mountain area of Sierra Nevada and surroundings. From these endemic taxa, 70% are endangered by different threats, global warming being a leading cause. We seek to provide a complete and updated database of the flora of the Sierra Nevada mountains (southeast Spain). The goal of the present data set is to compile the names of all the vascular plant taxa inhabiting Sierra Nevada, together with relevant features including taxonomical, morphological-ecological traits, distribution, habitats, abundance, and conservation status. The data were compiled according to all the available information sources on taxonomy, ecology, and plant-species distribution. The resulting data set includes 2,348 taxa belonging to 1,937 species, 377 subspecies, and 34 hybrids, from a total of 756 genera and 146 families represented in the collection. For each taxa, together with taxonomical information (Phylum, Class, Family, Genus, Taxa), we compiled plant traits (life form, spinescence, flower symmetry, flower sexuality, plant gender, androecium:ginoecium ratio, flower color, perianth type, pollinator type, flowering, seed dispersal, and vegetative reproduction), and their environmental association (origin, endemic character, general distribution, substrate, elevation, habitat, local abundance, hygrophilous behavior, and conservation status). All these traits were compiled from all the available information sources, resulting in a complete and updated database for Sierra Nevada vascular flora. This data set provides valuable information on plant traits in an outstanding micro hotspot within the Mediterranean hotspot. This data set can be freely used for noncommercial purposes. This data set is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0). When you use this data set, we request that you cite the data and this data paper.

Genetic diversity and differentiation in narrow versus widespread taxa of Helianthemum (Cistaceae) in a hotspot: The role of geographic range, habitat, and reproductive traits.

Unraveling the relationships between ecological, functional traits and genetic diversity of narrow endemic plants provide opportunities for understanding how evolutionary processes operate over local spatial scales and ultimately how diversity is created and maintained. To explore these aspects in Sierra Nevada, the core of the Mediterranean Betic-Rifean hotspot, we have analyzed nuclear DNA microsatellite diversity and a set of biological and environmental factors (physicochemical soil parameters, floral traits, and community composition) in two strictly endemic taxa from dolomite outcrops of Sierra Nevada (Helianthemum pannosum and H. apenninum subsp. estevei) and two congeneric widespread taxa (H. cinereum subsp. rotundifolium and H. apenninum subsp. apenninum) that further belong to two different lineages (subgenera) of Helianthemum. We obtained rather unexpected results contrasting with the theory: (a) The narrow endemic taxa showed higher values of genetic diversity as well as higher average values of pollen production per flower and pollen-to-ovule ratio than their widespread relatives; and (b) the two taxa of subg. Helianthemum, with larger corollas, approach herkogamy and higher pollen production than the two taxa of subg. Plectolobum, displayed lower genetic diversity and higher values of inbreeding. Altogether, these results disclose how genetic diversity may be affected simultaneously by a large number of intrinsic and extrinsic factors, especially in Pleistocene glacial refugia in mountains where the spatial context harbors a great ecological heterogeneity. On the other hand, differences in mating system and the significant effect of the substrate profile, both being highly diverse in the genus Helianthemum, in the genetic variability illustrate about the importance of these two factors in the diversification and species differentiation of this paradigmatic genus in the Mediterranean and open the field to formulate and test new hypotheses of local adaptation, trait evolution, and habitat diversification.

Molecular phylogeny and evolutionary history of Moricandia DC (Brassicaceae).

BACKGROUND: The phylogeny of tribe Brassiceae (Brassicaceae) has not yet been resolved because of its complex evolutionary history. This tribe comprises economically relevant species, including the genus Moricandia DC. This genus is currently distributed in North Africa, Middle East, Central Asia and Southern Europe, where it is associated with arid and semi-arid environments. Although some species of Moricandia have been used in several phylogenetic studies, the phylogeny of this genus is not well established. METHODS: Here we present a phylogenetic analysis of the genus Moricandia using a nuclear (the internal transcribed spacers of the ribosomal DNA) and two plastidial regions (parts of the NADH dehydrogenase subunit F gene and the trnT-trnF region). We also included in the analyses members of their sister genus Rytidocarpus and from the close genus Eruca. RESULTS: The phylogenetic analyses showed a clear and robust phylogeny of the genus Moricandia. The Bayesian inference tree was concordant with the maximum likelihood and timing trees, with the plastidial and nuclear trees showing only minor discrepancies. The genus Moricandia appears to be formed by two main lineages: the Iberian clade including three species, and the African clade including the four species inhabiting the Southern Mediterranean regions plus M. arvensis. DISCUSSION: We dated the main evolutionary events of this genus, showing that the origin of the Iberian clade probably occurred after a range expansion during the Messinian period, between 7.25 and 5.33 Ma. In that period, an extensive African-Iberian floral and faunal interchange occurred due to the existence of land bridges between Africa and Europa in what is, at present-days, the Strait of Gibraltar. We have demonstrated that a Spanish population previously ascribed to Rytidocarpus moricandioides is indeed a Moricandia species, and we propose to name it as M. rytidocarpoides sp. nov. In addition, in all the phylogenetic analyses, M. foleyi appeared outside the Moricandia lineage but within the genus Eruca. Therefore, M. foleyi should be excluded from the genus Moricandia and be ascribed, at least provisionally, to the genus Eruca.

Rock climbing alters plant species composition, cover, and richness in Mediterranean limestone cliffs.

Rock climbing is among the outdoor activities that have undergone the highest growth since the second half of the 20th century. As a result, cliff habitats, historically one of the least disturbed by human colonization worldwide, are facing more intense human pressure than ever before. However, there is little data on the impact of this activity in plant-communities, and such information is indispensable for adequate manager decision-making. The goal of this study was to determine the impact of rock climbing on plant communities in terms of cover, richness, and composition in relation to climbing intensity on typical Mediterranean limestone cliffs. Three rock-climbing sites were selected in the Baetic range (SE Spain), corresponding to qualitative categories of climbing frequentation: i)"low" (low frequentation with intermittent climbing), ii)"medium" (high frequentation without overcrowding), and iii) "high" (high frequentation with overcrowding). Within each site, we selected climbing routes and adjacent areas free of climbing, then we carried out a photoplot-based sampling by rappelling. We analysed the images to calculate: richness, species cover, and total cover. This study shows that rock climbing negatively affected the cliff plant community at all three study sites. A significant decrease in plant cover, species richness and a shift in the community composition were recorded for climbed areas, the cover being the variable most sensitive to rock climbing. Impact observed proved to be related to the frequentation level. Low-frequentation sites, with usually more specialized climbers, underwent relatively mild damages, whereas at high frequentation sites the impact was severe and the conservation of the species, especially rare ones, became jeopardized. Our study is the first one available to investigate climbing impact on plant communities in Mediterranean areas, but more research on the impact of rock climbing is needed to assess the regulation of this activity. Regarding management guidelines, we propose a management guideline protocol to evaluate climbing routes and design: i) "Sites free of climbing", ii) "Strictly regulated climbing routes", iii) "Mildly regulated climbing routes", or iv) "Free climbing routes".

Secondary coenzyme Q10 deficiencies in oxidative phosphorylation (OXPHOS) and non-OXPHOS disorders.

We evaluated the coenzyme Q10 (CoQ) levels in patients who were diagnosed with mitochondrial oxidative phosphorylation (OXPHOS) and non-OXPHOS disorders (n=72). Data from the 72 cases in this study revealed that 44.4% of patients showed low CoQ concentrations in either their skeletal muscle or skin fibroblasts. Our findings suggest that secondary CoQ deficiency is a common finding in OXPHOS and non-OXPHOS disorders. We hypothesize that cases of CoQ deficiency associated with OXPHOS defects could be an adaptive mechanism to maintain a balanced OXPHOS, although the mechanisms explaining these deficiencies and the pathophysiological role of secondary CoQ deficiency deserves further investigation.

Designing conservation strategies to preserve the genetic diversity of Astragalus edulis Bunge, an endangered species from western Mediterranean region.

Astragalus edulis (Fabaceae) is an endangered annual species from the western Mediterranean region that colonized the SE Iberian Peninsula, NE and SW Morocco, and the easternmost Macaronesian islands (Lanzarote and Fuerteventura). Although in Spain some conservation measures have been adopted, it is still necessary to develop an appropriate management plan to preserve genetic diversity across the entire distribution area of the species. Our main objective was to use population genetics as well as ecological and phylogeographic data to select Relevant Genetic Units for Conservation (RGUCs) as the first step in designing conservation plans for A. edulis. We identified six RGUCs for in situ conservation, based on estimations of population genetic structure and probabilities of loss of rare alleles. Additionally, further population parameters, i.e. occupation area, population size, vulnerability, legal status of the population areas, and the historical haplotype distribution, were considered in order to establish which populations deserve conservation priority. Three populations from the Iberian Peninsula, two from Morocco, and one from the Canary Islands represent the total genetic diversity of the species and the rarest allelic variation. Ex situ conservation is recommended to complement the preservation of A. edulis, given that effective in situ population protection is not feasible in all cases. The consideration of complementary phylogeographic and ecological data is useful for management efforts to preserve the evolutionary potential of the species.

The role of pollinators in the evolution of corolla shape variation, disparity and integration in a highly diversified plant family with a conserved floral bauplan.

BACKGROUND AND AIMS: Brassicaceae is one of the most diversified families in the angiosperms. However, most species from this family exhibit a very similar floral bauplan. In this study, we explore the Brassicaceae floral morphospace, examining how corolla shape variation (an estimation of developmental robustness), integration and disparity vary among phylogenetically related species. Our aim is to check whether these floral attributes have evolved in this family despite its apparent morphological conservation, and to test the role of pollinators in driving this evolution. METHODS: Using geometric morphometric tools, we calculated the phenotypic variation, disparity and integration of the corolla shape of 111 Brassicaceae taxa. We subsequently inferred the phylogenetic relationships of these taxa and explored the evolutionary lability of corolla shape. Finally, we sampled the pollinator assemblages of every taxon included in this study, and determined their pollination niches using a modularity algorithm. We explore the relationship between pollination niche and the attributes of corolla shape. KEY RESULTS: Phylogenetic signal was weak for all corolla shape attributes. All taxa had generalized pollination systems. Nevertheless, they belong to different pollination niches. There were significant differences in corolla shape among pollination niches even after controlling for the phylogenetic relationship of the plant taxa. Corolla shape variation and disparity was significantly higher in those taxa visited mostly by nocturnal moths, indicating that this pollination niche is associated with a lack of developmental robustness. Corolla integration was higher in those taxa visited mostly by hovering long-tongued flies and long-tongued large bees. CONCLUSIONS: Corolla variation, integration and disparity were evolutionarily labile and evolved very recently in the evolutionary history of the Brassicaceae. These floral attributes were strongly related to the pollination niche. Even in a plant clade having a very generalized pollination system and exhibiting a conserved floral bauplan, pollinators can drive the evolution of important developmental attributes of corolla shape.

[Guidelines for monitoring late-onset Pompe disease.Sociedad Española de Medicina Interna (SEMI), Sociedad Española de Neurología (SEN) y Sociedad Española de Neumología y CirugíaTorácica (SEPAR)]. / Guía para el seguimiento de la enfermedad de Pompe de inicio tardío.

Although treatment with alglucosidase alfa has helped improve the prognosis of patients with late-onset Pompe disease, both the development of the disease and the effectiveness of the treatment need to be monitored on a regular basis. This is the reason that has led a committee of Spanish experts to draw up a series of guidelines on how to follow up these patients. The committee proposes a model of follow-up tests for late-onset Pompe disease. First of all, the nutritional status and swallowing function must be evaluated. Second, and due to the variability of the clinical features, the committee recommends the simultaneous use of several scales to measure different functions and parameters. Thus, muscular force is assessed with the Medical Research Council scale; motor functioning, with the six-minute walk test and timed tests; disability, with the Rasch-built Pompe-specific Activity scale; respiratory functioning, with measurement of the forced vital capacity and oxygen saturation; and fatigue, with the fatigue intensity scale. Lastly, the safety and tolerability of enzyme replacement therapy are controlled by registering and treating the potential side effects and measurement of the anti-alglucosidase alfa antibodies. A number of different general recommendations are also included.

TITLE: Guia para el seguimiento de la enfermedad de Pompe de inicio tardio.Aunque el tratamiento con alglucosidasa alfa ha contribuido a mejorar el pronostico de los pacientes con enfermedad de Pompe de inicio tardio, es necesario hacer un seguimiento periodico de la evolucion de la enfermedad y de la eficacia del tratamiento. Por este motivo, un comite de expertos españoles ha elaborado una guia para el seguimiento de estos pacientes. El comite propone un modelo de pruebas de seguimiento para la enfermedad de Pompe de inicio tardio. En primer lugar, ha de valorarse el estado nutricional y la funcion deglutoria. En segundo lugar, y debido a la variabilidad del cuadro clinico, el comite recomienda el uso simultaneo de varias escalas que midan distintas funciones y parametros. De este modo, la fuerza muscular se evalua con la escala del Medical Research Council; la funcion motora, con la prueba de la marcha en seis minutos y pruebas cronometradas; la discapacidad, con la escala de actividad especifica de la enfermedad de Pompe construida segun el analisis de Rasch; la funcion respiratoria, con la medida de la capacidad vital forzada y la saturacion de oxigeno; y la fatiga, con la escala de intensidad de la fatiga. Por ultimo, la seguridad y la tolerabilidad del tratamiento enzimatico sustitutivo se controlan con el registro y tratamiento de los potenciales efectos adversos y la medicion de los anticuerpos antialglucosidasa alfa. Se incluyen tambien diversas recomendaciones generales.

The role of pollinators in floral diversification in a clade of generalist flowers.

Pollinator-mediated evolutionary divergence has seldom been explored in generalist clades because it is assumed that pollinators in those clades exert weak and conflicting selection. We investigate whether pollinators shape floral diversification in a pollination generalist plant genus, Erysimum. Species from this genus have flowers that appeal to broad assemblages of pollinators. Nevertheless, we recently reported that it is possible to sort plant species into pollination niches varying in the quantitative composition of pollinators. We test here whether floral characters of Erysimum have evolved as a consequence of shifts among pollination niches. For this, we quantified the evolutionary lability of the floral traits and their phylogenetic association with pollination niches. As with pollination niches, Erysimum floral traits show weak phylogenetic signal. Moreover, floral shape and color are phylogenetically associated with pollination niche. In particular, plants belonging to a pollination niche dominated by long-tongued large bees have lilac corollas with parallel petals. Further analyses suggest, however, that changes in color preceded changes in pollination niche. Pollinators seem to have driven the evolution of corolla shape, whereas the association between pollination niche and corolla color has probably arisen by lilac-flowered Erysimum moving toward certain pollination niches for other adaptive reasons.

Evolution of pollination niches in a generalist plant clade.

It is widely assumed that floral diversification occurs by adaptive shifts between pollination niches. In contrast to specialized flowers, identifying pollination niches of generalist flowers is a challenge. Consequently, how generalist pollination niches evolve is largely unknown. We apply tools from network theory and comparative methods to investigate the evolution of pollination niches among generalist species belonging to the genus Erysimum. These species have similar flowers. We found that the studied species may be grouped in several multidimensional niches separated not by a shift of pollinators, but instead by quantitative variation in the relative abundance of pollinator functional groups. These pollination niches did not vary in generalization degree; we did not find any evolutionary trend toward specialization within the studied clade. Furthermore, the evolution of pollination niche fitted to a Brownian motion model without phylogenetic signal, and was characterized by frequent events of niche convergences and divergences. We presume that the evolution of Erysimum pollination niches has occurred mostly by recurrent shifts between slightly different generalized pollinator assemblages varying spatially as a mosaic and without any change in specialization degree. Most changes in pollination niches do not prompt floral divergence, a reason why adaptation to pollinators is uncommon in generalist plants.

Sinfonevada: Dataset of Floristic diversity in Sierra Nevada forests (SE Spain).

The Sinfonevada database is a forest inventory that contains information on the forest ecosystem in the Sierra Nevada mountains (SE Spain). The Sinfonevada dataset contains more than 7,500 occurrence records belonging to 270 taxa (24 of these threatened) from floristic inventories of the Sinfonevada Forest inventory. Expert field workers collected the information. The whole dataset underwent a quality control by botanists with broad expertise in Sierra Nevada flora. This floristic inventory was created to gather useful information for the proper management of Pinus plantations in Sierra Nevada. This is the only dataset that shows a comprehensive view of the forest flora in Sierra Nevada. This is the reason why it is being used to assess the biodiversity in the very dense pine plantations on this massif. With this dataset, managers have improved their ability to decide where to apply forest treatments in order to avoid biodiversity loss. The dataset forms part of the Sierra Nevada Global Change Observatory (OBSNEV), a long-term research project designed to compile socio-ecological information on the major ecosystem types in order to identify the impacts of global change in this area.

Post-fire salvage logging alters species composition and reduces cover, richness, and diversity in Mediterranean plant communities.

An intense debate exists on the effects of post-fire salvage logging on plant community regeneration, but scant data are available derived from experimental studies. We analyzed the effects of salvage logging on plant community regeneration in terms of species richness, diversity, cover, and composition by experimentally managing a burnt forest on a Mediterranean mountain (Sierra Nevada, S Spain). In each of three plots located at different elevations, three replicates of three treatments were implemented seven months after the fire, differing in the degree of intervention: "Non-Intervention" (all trees left standing), "Partial Cut plus Lopping" (felling 90% of the trees, cutting the main branches, and leaving all the biomass in situ), and "Salvage Logging" (felling and piling the logs, and masticating the woody debris). Plant composition in each treatment was monitored two years after the fire in linear point transects. Post-fire salvage logging was associated with reduced species richness, Shannon diversity, and total plant cover. Moreover, salvaged sites hosted different species assemblages and 25% lower cover of seeder species (but equal cover of resprouters) compared to the other treatments. Cover of trees and shrubs was also lowest in Salvage Logging, which could suggest a potential slow-down of forest regeneration. Most of these results were consistent among the three plots despite plots hosting different plant communities. Concluding, our study suggests that salvage logging may reduce species richness and diversity, as well as the recruitment of woody species, which could delay the natural regeneration of the ecosystem.

Reduction of pain thresholds in fibromyalgia after very low-intensity magnetic stimulation: a double-blinded, randomized placebo-controlled clinical trial.

BACKGROUND: Exposure to electromagnetic fields has been reported to have analgesic and antinociceptive effects in several organisms. OBJECTIVE: To test the effect of very low-intensity transcranial magnetic stimulation on symptoms associated with fibromyalgia syndrome. METHODS: A double-blinded, placebo-controlled clinical trial was performed in the Sagrado Corazón Hospital, Seville, Spain. Female fibromyalgia patients (22 to 50 years of age) were randomly assigned to either a stimulation group or a sham group. The stimulation group (n=28) was stimulated using 8 Hz pulsed magnetic fields of very low intensity, while the sham group (n=26) underwent the same protocol without stimulation. Pressure pain thresholds before and after stimulation were determined using an algometer during the eight consecutive weekly sessions of the trial. In addition, blood serotonin levels were measured and patients completed questionnaires to monitor symptom evolution. RESULTS: A repeated-measures ANOVA indicated statistically significant improvement in the stimulation group compared with the control group with respect to somatosensory pain thresholds, ability to perform daily activities, perceived chronic pain and sleep quality. While improvement in pain thresholds was apparent after the first stimulation session, improvement in the other three measures occurred after the sixth week. No significant between-group differences were observed in scores of depression, fatigue, severity of headaches or serotonin levels. No adverse side effects were reported in any of the patients. CONCLUSIONS: Very low-intensity magnetic stimulation may represent a safe and effective treatment for chronic pain and other symptoms associated with fibromyalgia.

Screening of effective pharmacological treatments for MELAS syndrome using yeasts, fibroblasts and cybrid models of the disease.

BACKGROUND AND PURPOSE: MELAS (mitochondrial encephalomyopathy, lactic acidosis and stroke-like episodes) is a mitochondrial disease most usually caused by point mutations in tRNA genes encoded by mitochondrial DNA (mtDNA). Approximately 80% of cases of MELAS syndrome are associated with a m.3243A > G mutation in the MT-TL1 gene, which encodes the mitochondrial tRNALeu (UUR). Currently, no effective treatments are available for this chronic progressive disorder. Treatment strategies in MELAS and other mitochondrial diseases consist of several drugs that diminish the deleterious effects of the abnormal respiratory chain function, reduce the presence of toxic agents or correct deficiencies in essential cofactors. EXPERIMENTAL APPROACH: We evaluated the effectiveness of some common pharmacological agents that have been utilized in the treatment of MELAS, in yeast, fibroblast and cybrid models of the disease. The yeast model harbouring the A14G mutation in the mitochondrial tRNALeu(UUR) gene, which is equivalent to the A3243G mutation in humans, was used in the initial screening. Next, the most effective drugs that were able to rescue the respiratory deficiency in MELAS yeast mutants were tested in fibroblasts and cybrid models of MELAS disease. KEY RESULTS: According to our results, supplementation with riboflavin or coenzyme Q(10) effectively reversed the respiratory defect in MELAS yeast and improved the pathologic alterations in MELAS fibroblast and cybrid cell models. CONCLUSIONS AND IMPLICATIONS: Our results indicate that cell models have great potential for screening and validating the effects of novel drug candidates for MELAS treatment and presumably also for other diseases with mitochondrial impairment.