Investigating the Effectiveness of a Carb-Free Oloproteic Diet in Fibromyalgia Treatment.

Fibromyalgia (FM), a chronic disease with a high incidence in women, poses a significant challenge for diagnosis and treatment, especially due to the absence of specific biomarkers and the multifaceted nature of its symptoms, which range from neuromuscular pain to mood disorders and intestinal dysbiosis. While diagnosis currently relies on rheumatological clinical evaluations and treatment options mainly focus on symptom management, FM seems to have possible links with systemic metabolic dysfunctions with a common inflammatory root. In this context, a new therapeutic avenue emerges: could a therapeutic nutritional approach be the missing piece of the puzzle? Indeed, diet therapies employed particularly for metabolic syndromes proved recently to be efficacious for correcting systemic dysmetabolism and a high number of chronic inflammation conditions. In particular, the very-low-calorie ketogenic diet (VLCKD) demonstrated therapeutic benefits in many disorders. In the present study, we aimed to investigate the specific effects of two dietary interventions, namely the oloproteic VLCKD and the low-glycemic insulinemic (LOGI) diet, on two groups of female FM patients (FM1 and FM2) over a 45-day period. Utilizing clinical and laboratory tests, as well as non-invasive NMR metabolomic analysis of serum, urine, and saliva samples, we sought to uncover how these dietary regimens impact the metabolic dysfunctions associated with FM.

Acceso para cirugía de catarata en diabetes. Un estudio cualitativo con enfoque gerencial / Access for cataract surgery in patients with diabetes. A qualitative study with a managerial approach

Resumen Objetivo: Explorar las brechas que existen para el acceso a la cirugía de catarata en personas con diabetes tipo 2. Materiales y métodos: Se realizó una investigación cualitativa de tipo fenomenológica con enfoque gerencial. Se emplearon herramientas y técnicas de gestión por procesos en salud y metodología Lean- sanidad. El trabajo de campo se realizó en los municipios La Lisa, Marianao y Playa donde se exploraron las experiencias y expectativas de 46 pacientes en seis grupos de discusión, se obtuvo el criterio de nueve médicos de familia y seis oftalmólogos del nivel primario de salud, en dos grupos de discusión. Resultados: Las experiencias de pacientes, médicos de familia y oftalmólogos, organizadas por categorías (diagnóstico, continuidad y comunicación, apoyo social y carga laboral) tienen en su contenido las brechas presentes en el proceso asistencial. Conclusión: Las brechas identificadas en este estudio son suficientes para explicar por qué muchos pacientes no acceden de manera oportuna a la cirugía de catarata.

Abstract Objective: To explore the gaps that exist for access to cataract surgery in people with type 2 diabetes. Materials and methods: A qualitative research of a phenomenological type with a managerial approach was carried out. Management tools and techniques by health processes and Lean-health methodology were used. The field work was carried out in the municipalities of La Lisa, Marianao and Playa where the experiences and expectations of 46 patients were explored in six discussion groups, the criteria of nine family doctors and six ophthalmologists from the primary health level were obtained, in two discussion groups. Results: The experiences of patients, family doctors and ophthalmologists, organized by categories (diagnosis, continuity and communication, social support and workload) have in their content the gaps present in the care process. Conclusion: The gaps identified in this study are sufficient to explain why many patients do not access cataract surgery in a timely manner.

Exploiting the Features of Short Peptides to Recognize Specific Cell Surface Markers.

Antibodies are the macromolecules of choice to ensure specific recognition of biomarkers in biological assays. However, they present a range of shortfalls including a relatively high production cost and limited tissue penetration. Peptides are relatively small molecules able to reproduce sequences of highly specific paratopes and, although they have less biospecificity than antibodies, they offer advantages like ease of synthesis, modifications of their amino acid sequences and tagging with fluorophores and other molecules required for detection. This work presents a strategy to design peptide sequences able to recognize the CD44 hyaluronic acid receptor present in the plasmalemma of a range of cells including human bone marrow stromal mesenchymal cells. The protocol of identification of the optimal amino acid sequence was based on the combination of rational design and in silico methodologies. This protocol led to the identification of two peptide sequences which were synthesized and tested on human bone marrow mesenchymal stromal cells (hBM-MSCs) for their ability to ensure specific binding to the CD44 receptor. Of the two peptides, one binds CD44 with sensitivity and selectivity, thus proving its potential to be used as a suitable alternative to this antibody in conventional immunostaining. In the context of regenerative medicine, the availability of this peptide could be harnessed to functionalize tissue engineering scaffolds to anchor stem cells as well as to be integrated into systems such as cell sorters to efficiently isolate MSCs from biological samples including various cell subpopulations. The data here reported can represent a model for developing peptide sequences able to recognize hBM-MSCs and other types of cells and for their integration in a range of biomedical applications.

Supplementing Low-Sodium Bicarbonate-Calcic (Lete)® Water: Effects in Women on Bone and Systemic Metabolism.

Calcium (Ca) represents about 40% of the total mineral mass, mainly in the bone, providing mechanical strength to the skeleton and teeth. An adequate Ca intake is necessary for bone growth and development in children and adolescents and for maintaining bone mineral loss in elderly age. Ca deficiency predisposes to osteopenia and osteoporosis. Healthy nutrition, including an adequate intake of Ca-rich food, is paramount to prevent and cure osteoporosis. Recently, several clinical studies have demonstrated that, in conditions of Ca dysmetabolism, Ca-rich mineral water is beneficial as a valuable source of Ca to be used as an alternative to caloric Ca-rich dairy products. Although promising, these data have been collected from small groups of participants. Moreover, they mainly regard the effect of Ca-rich mineral water on bone metabolism. In contrast, an investigation of the effect of Ca supplementation on systemic metabolism is needed to address the spreading of systemic metabolic dysfunction often associated with Ca dysmetabolism. In the present study, we analyzed urine and blood sera of 120 women in perimenopausal condition who were subjected for six months to 2l daily consumption of bicarbonate-calcium mineral water marketed under ®Lete. Remarkably, this water, in addition to being rich in calcium and bicarbonate, is also low in sodium. A complete set of laboratory tests was carried out to investigate whether the specific water composition was such to confirm the known therapeutic effects on bone metabolism. Second, but not least, urine and blood sera were analyzed using NMR-based metabolomic procedures to investigate, other than the action on Ca metabolism, potential system-wide metabolic effects. Our data show that Lete water is a valid supplement for compensating for Ca dysmetabolism and preserving bone health and integrity.

Lipophilic Shellfish Poisoning Toxins in Marine Invertebrates from the Galician Coast.

For the purpose of assessing human health exposure, it is necessary to characterize the toxins present in a given area and their potential impact on commercial species. The goal of this research study was: (1) to screen the prevalence and concentrations of lipophilic toxins in nine groups of marine invertebrates in the northwest Iberian Peninsula; (2) to evaluate the validity of wild mussels (Mytilus galloprovincialis) as sentinel organisms for the toxicity in non-bivalve invertebrates from the same area. The screening of multiple lipophilic toxins in 1150 samples has allowed reporting for the first time the presence of 13-desmethyl spirolide C, pinnatoxin G, okadaic acid, and dinophysistoxins 2 in a variety of non-traditional vectors. In general, these two emerging toxins showed the highest prevalence (12.5-75%) in most of the groups studied. Maximum levels for 13-desmethyl spirolide C and pinnatoxin G were found in the bivalves Magallana gigas (21 µg kg-1) and Tellina donacina (63 µg kg-1), respectively. However, mean concentrations for the bivalve group were shallow (2-6 µg kg-1). Okadaic acid and dinophysistoxin 2 with lower prevalence (1.6-44.4%) showed, on the contrary, very high concentration values in specific species of crustaceans and polychaetes (334 and 235 µg kg--1, respectively), to which special attention should be paid. Statistical data analyses showed that mussels could be considered good biological indicators for the toxicities of certain groups in a particular area, with correlations between 0.710 (for echinoderms) and 0.838 (for crustaceans). Polychaetes could be an exception, but further extensive surveys would be needed to draw definitive conclusions.

SMN deficiency perturbs monoamine neurotransmitter metabolism in spinal muscular atrophy.

Beyond motor neuron degeneration, homozygous mutations in the survival motor neuron 1 (SMN1) gene cause multiorgan and metabolic defects in patients with spinal muscular atrophy (SMA). However, the precise biochemical features of these alterations and the age of onset in the brain and peripheral organs remain unclear. Using untargeted NMR-based metabolomics in SMA mice, we identify cerebral and hepatic abnormalities related to energy homeostasis pathways and amino acid metabolism, emerging already at postnatal day 3 (P3) in the liver. Through HPLC, we find that SMN deficiency induces a drop in cerebral norepinephrine levels in overt symptomatic SMA mice at P11, affecting the mRNA and protein expression of key genes regulating monoamine metabolism, including aromatic L-amino acid decarboxylase (AADC), dopamine beta-hydroxylase (DßH) and monoamine oxidase A (MAO-A). In support of the translational value of our preclinical observations, we also discovered that SMN upregulation increases cerebrospinal fluid norepinephrine concentration in Nusinersen-treated SMA1 patients. Our findings highlight a previously unrecognized harmful influence of low SMN levels on the expression of critical enzymes involved in monoamine metabolism, suggesting that SMN-inducing therapies may modulate catecholamine neurotransmission. These results may also be relevant for setting therapeutic approaches to counteract peripheral metabolic defects in SMA.

First Report of Two Gymnodimines and Two Tetrodotoxin Analogues in Invertebrates from the North Atlantic Coast of Spain.

Gymnodimine D (GYM D), 16-desmethyl gymnodimine D (16-desmethyl GYM D), and two tetrodotoxin analogues have been found in invertebrates obtained from the north Atlantic coast of Spain from May 2021 to October 2022. It is the first report of GYMD and 16-desmethyl GYM D in invertebrates worldwide and of the tetrodotoxin analogues, 5,6,11 trideoxy tetrodotoxin (5,6,11 trideoxy TTX) and its isomer (referred to as 5,6,11 trideoxy-epi-TTX), in the north Atlantic Coast of Spain. In this study, we also report for the first time the detection of tetrodotoxin (TTX) in three species (the cnidaria Calliactis parasitica, an unidentified species, and the bivalve Tellina donacina). The prevalence was medium for GYM D and 16-desmethyl GYM D and low for TTXs overall. The concentrations recorded were variable, with maximum values of GYM D in the bivalve Cerastoderma edule (8.8 µg GYM A equivalents kg-1), of 16-desmethyl GYM D in the bivalve Magellana gigas (10 µg GYM A equivalents kg-1) and of TTX and 5,6,11 trideoxy TTX in the cnidaria C. parasitica (49.7 and 233 µg TTX equivalents kg-1, respectively). There is very scarce information about these compounds. Therefore, the reporting of these new detections will increase the knowledge on the current incidence of marine toxins in Europe that the European Food Safety Authority (EFSA), in particular, and the scientific community, in general, have. This study also highlights the importance of analyzing toxin analogues and metabolites for effective monitoring programs and adequate health protection.

Determinantes externos de la disponibilidad de sangre / External Determinants of Blood Availability

El acceso universal a las transfusiones de sangre depende de diversos factores localizados dentro y fuera de los servicios de salud. Motivados por ser este un tema infrecuente, el trabajo tiene el objetivo de analizar desde una perspectiva multidimensional y holística los determinantes externos de la disponibilidad de sangre para las transfusiones. El desarrollo socioeconómico constituye un importante factor dinamizador porque condiciona, de cierta manera, la infraestructura del sistema sanitario e incide sobre los factores epidemiológicos, sociodemográficos y jurídicos. A su vez, estos influyen en la organización de las actividades y la complejidad de la asistencia sanitaria, factores que otorgan un sentido positivo o negativo a la disponibilidad de sangre. Tomarlos en consideración al diseñar las políticas de salud relacionadas con el uso de la sangre permite una mayor precisión en las intervenciones locales que contribuyan a mejorar la seguridad de las transfusiones(AU)

Universal access to blood transfusions depends on a number of localized factors both inside and outside health services. Motivated by being an infrequent topic, this work aims to analyze from a multidimensional and holistic perspective the external determinants of the availability of blood for transfusions. Socio-economic development is an important dynamic factor because it conditions, in a certain way, the infrastructure of the health system and affects epidemiological, sociodemographic and legal factors. In turn, these influence the organization of activities and the complexity of health care, factors that give a positive or negative meaning to the availability of blood. Taking them into consideration when designing health policies related to the use of blood allows greater precision in local interventions that contribute to improving the safety of transfusions(AU)

Enzymatic Biotransformation of 13-desmethyl Spirolide C by Two Infaunal Mollusk Species: The Limpet Patella vulgata and the Cockle Cerastoderma edule.

The presence of a 13-desmethyl Spirolide C isomer (Iso-13-desm SPX C) is very common in some infaunal mollusks in Galicia contaminated with this toxin. Its possible origin by biological transformation was investigated by incubating homogenates of the soft tissues of limpets and cockles spiked with 13-desmethyl Spirolide C (13-desm SPX C). The involvement of an enzymatic process was also tested using a raw and boiled cockle matrix. The enzymatic biotransformation of the parent compound into its isomer was observed in the two species studied, but with different velocities. The structural similarity between 13-desm SPX C and its isomer suggests that epimerization is the most likely chemical process involved. Detoxification of marine toxins in mollusks usually implies the enzymatic biotransformation of original compounds, such as hydroxylation, demethylation, or esterification; however, this is the first time that this kind of transformation between spirolides in mollusks has been demonstrated.

Nusinersen Induces Disease-Severity-Specific Neurometabolic Effects in Spinal Muscular Atrophy.

Intrathecal delivery of Nusinersen-an antisense oligonucleotide that promotes survival motor neuron (SMN) protein induction-is an approved therapy for spinal muscular atrophy (SMA). Here, we employed nuclear magnetic resonance (NMR) spectroscopy to longitudinally characterize the unknown metabolic effects of Nusinersen in the cerebrospinal fluid (CSF) of SMA patients across disease severity. Modulation of amino acid metabolism is a common denominator of biochemical changes induced by Nusinersen, with distinct downstream metabolic effects according to disease severity. In severe SMA1 patients, Nusinersen stimulates energy-related glucose metabolism. In intermediate SMA2 patients, Nusinersen effects are also related to energy homeostasis but involve ketone body and fatty acid biosynthesis. In milder SMA3 patients, Nusinersen mainly modulates amino acid metabolism. Moreover, Nusinersen modifies the CSF metabolome of a more severe clinical group towards the profile of untreated SMA patients with milder disease. These findings reveal disease severity-specific neurometabolic signatures of Nusinersen treatment, suggesting a selective modulation of peripheral organ metabolism by this CNS-directed therapy in severe SMA patients.

The Metabolomic Profile in Amyotrophic Lateral Sclerosis Changes According to the Progression of the Disease: An Exploratory Study.

Amyotrophic lateral sclerosis (ALS) is a multifactorial neurodegenerative pathology of the upper or lower motor neuron. Evaluation of ALS progression is based on clinical outcomes considering the impairment of body sites. ALS has been extensively investigated in the pathogenetic mechanisms and the clinical profile; however, no molecular biomarkers are used as diagnostic criteria to establish the ALS pathological staging. Using the source-reconstructed magnetoencephalography (MEG) approach, we demonstrated that global brain hyperconnectivity is associated with early and advanced clinical ALS stages. Using nuclear magnetic resonance (1H-NMR) and high resolution mass spectrometry (HRMS) spectroscopy, here we studied the metabolomic profile of ALS patients' sera characterized by different stages of disease progression-namely early and advanced. Multivariate statistical analysis of the data integrated with the network analysis indicates that metabolites related to energy deficit, abnormal concentrations of neurotoxic metabolites and metabolites related to neurotransmitter production are pathognomonic of ALS in the advanced stage. Furthermore, analysis of the lipidomic profile indicates that advanced ALS patients report significant alteration of phosphocholine (PCs), lysophosphatidylcholine (LPCs), and sphingomyelin (SMs) metabolism, consistent with the exigency of lipid remodeling to repair advanced neuronal degeneration and inflammation.

Diabetes mellitus y salud visual como referentes esenciales en la práctica médica / Diabetes mellitus and visual health as essential references in medical practice

Introducción: La diabetes mellitus, requiere atención médica multifactorial y multisectorial. La oftalmopatía diabética demanda sistemas de salud integrados que faciliten la prevención, detección y tratamiento de las complicaciones en los niveles de atención primaria, secundaria y terciaria. Objetivo: Exponer los conceptos teóricos, prácticos y las tendencias actuales sobre diabetes mellitus y salud visual. Métodos: Se realizó una revisión bibliografía y documental sobre diabetes mellitus y salud visual, estudios y documentos publicados por instituciones nacionales e internacionales, temas vinculados a la diabetes mellitus y la salud visual, se accedió a artículos desde diferentes fuentes, revistas indexadas, metaanálisis y fuentes primarias no publicadas, tesis de maestría, y doctorados. Resultados: Para conservar la salud visual es necesario llevar a cabo medidas que conlleven a prevenir la pérdida de la visión por catarata, glaucoma, retinopatia y edema macular diabético, que incluyen no sólo un buen control de los parámetros metabólicos y las revisiones periódicas al oftalmólogo, sino también el cumplimiento de la educación diabetológica por un equipo multidisciplinario y actualización sobre este tema de los prestadores de salud. La disminución visual que provoca la catarata y otras enfermedades oculares puede conducir a padecer discapacidades como, pérdida de autonomía para administrarse los medicamentos, limitar las actividades de la vida cotidiana e incapacidad laboral y social, con la consiguiente afectación de la calidad de vida Conclusiones: Acudir al oftalmólogo con frecuencia, mantener el control glucémico, del colesterol y presión arterial representan el pilar fundamental para preservar la salud visual en el paciente diabético(AU)

Introduction: Diabetes mellitus requires multifactorial and multisectoral medical attention. Diabetic ophthalmopathy demands integrated health systems that facilitate the prevention, detection and treatment of complications at the primary, secondary and tertiary care levels. Objective: To show the theoretical and practical concepts and current trends on diabetes mellitus and visual health. Methods: A bibliographic and documentary review on diabetes mellitus and visual health was carried out. Studies and documents published by local and international institutions, articles, topics related to diabetes mellitus and visual health were retrieved from different sources, indexed journals, meta-analysis and unpublished primary sources, research degree and master's, theses. Results: To preserve visual health, it is necessary to carry out reading that lead to preventing vision loss due to cataract, glaucoma, retinopathy and diabetic macular edema, which include not only good control of metabolic parameters and regular check-ups with the ophthalmologist , but also compliance with diabetes education by a multidisciplinary team and updating the health providers on this subject. The visual decrease caused by cataract and other eye diseases can lead to disabilities such as loss of autonomy to manage medications, limit activities of daily life and work and social incapacity, with the consequent impact on quality of life. Conclusions: Frequently going to the ophthalmologist, maintaining glycemic, cholesterol and blood pressure control represent the fundamental pillar to preserve visual health in diabetic patients(AU)

Transformación integral del sistema de salud de la República Democrática de Timor Leste (2013-2016) / Comprehensive transformation of the health system of the Democratic Republic of Timor Leste (2013-2016)

Resumen Objetivo: Implementar un proyecto de reformas al sistema de salud de la República Democrática de Timor Leste, en el periodo 2013-2016. Materiales y métodos: Se realizó un estudio de tipo investigación/acción desde una perspectiva gerencial. Se incluyó muestra no probabilística por conveniencia de usuarios, proveedores y funcionarios, se emplearon técnicas cualitativas como grupos focales y entrevistas abiertas, así como el uso de herramientas gerenciales. Resultados: En una primera fase se realizó el análisis situacional a escala de país. Con estos elementos se definieron sucesivamente las prioridades estratégicas, los objetivos, indicadores y metas por cada perspectiva. Finalmente, el programa de cambios se organizó, se ejecutó el proceso de transformaciones y se fueron determinando los resultados. Se diseñaron y quedaron funcionando el Programa Nacional de Cáncer y el Programa Nacional de Atención Primaria; se introdujeron especialidades médicas y servicios asistenciales con los que no se contaba; se crearon nuevos enfoques para la gestión de los recursos humanos y, se elevó el nivel del trabajo científico, entre otros beneficios. Conclusiones: Se produjeron mejoras sustanciales en el Sistema de Salud y además se dejó creada una capacidad potencial para que dichos efectos sigan creciendo en cantidad y calidad.

Abstract Objective: Implement a Project to reform the health system of the Democratic Republic of Timor Leste in the period 2013-2016. Materials and methods: A research / action type study was carried out from a managerial perspective. A non-probabilistic sample was included for the convenience of users, suppliers and officials, qualitative techniques such as focus groups and open interviews were used, as well as the use of management tools. Results: In a first phase, the situational analysis was carried out at the country level. With these elements, the strategic priorities, objectives, indicators and goals were successively defined for each perspective. Finally, the change program was organized, the transformation process was carried out and the results were determined. The National Cancer Program and the National Primary Care Program were designed and kept functioning; Missing medical specialties and care services were introduced; New approaches to human resource management were created and the level of scientific work was raised, among other benefits. Conclusions: There were substantial improvements in the health system and in addition, a potential capacity was created so that these effects continue to grow in quantity and quality.

Okadaic Acid Depuration from the Cockle Cerastoderma edule.

The cockle Cerastoderma edule is a commercially important species in many European Countries. It can accumulate okadaic acid (OA) and other toxins in its group, which makes it unsuitable for human consumption, producing harvesting bans to avoid intoxications. The duration of those bans depends in part on the depuration kinetics of the toxin in this species. In this work, this kinetics was studied by means of fitting different models to depuration data experimentally obtained, using naturally contaminated cockles. Cockles depurated OA faster than most other bivalve species studied. Models that include Michaelis-Menten kinetics describe the depuration better than those using a first order exponential decrease to describe the first (or the only) compartment. One-compartment models were not able to describe the final part of the depuration curve, in which OA was depurated very slowly. Therefore, two-compartment models were needed. Esters were depurated at a much faster rate than the free form of the toxin; however, no significant esterification was detected during the process. The slow depuration rate suggests that other bivalve species could be used as sentinels to monitor cockle populations, but caution should be taken when toxin concentrations are very high.

Duración de lactancia materna y consumo de productos ultraprocesados y bebidas azucaradas en niños uruguayos menores de 4 años

RESUMEN Se ha descrito que una mayor duración de lactancia materna, parece estar asociada a hábitos alimentarios saludables en la primera infancia. Nuestro objetivo fue determinar la asociación entre duración de lactancia materna y consumo de productos ultra-procesados (PUP) y bebidas azucaradas (BA) en menores de 4 años, considerando seguridad alimentaria y contexto educativo. Adicionalmente se analizó ingesta de otros alimentos. Se analizaron transversalmente 3.077 niños de la Primera Encuesta Nacional de Salud, Nutrición y Desarrollo Infantil de Uruguay. Se aplicaron cuestionarios a la madre u otro adulto responsable. Se realizaron modelos de regresión logística crudos y ajustados por: edad materna, índice de masa corporal materna al momento de la entrevista, seguridad alimentaria y asistencia a centro educativo. Se consideraron significativos valores de p<0.05. En modelos ajustados no se encontró asociación entre duración de lactancia materna y PUP, ni BA (p≥0.31) en ningún grupo etario. En el total de niños ≥6 meses se encontró asociación significativa directa entre inseguridad alimentaria y consumo de PUP y BA (p<0.046). En niños ≥24 meses, se observó asociación inversa entre asistencia a centro educativo y BA (p= 0.002). Los ≥24 meses en la categoría menos saludable (BA SI+0-4 alimentos saludables), nunca recibieron lactancia (p= 0.001); y en la más saludable (BA NO+4-8 alimentos saludables) el 72.4% había tenido lactancia ≥6 meses (p= 0.001). En conclusión, queda en evidencia que la inseguridad alimentaria explica en gran medida el consumo tanto de alimentos ultra-procesados como de bebidas azucaradas.

ABSTRACT Longer duration of breastfeeding seems to be associated with healthier eating habits in children. The aim of this study was to determine the association between breastfeeding duration and the consumption of ultra-processed products and sweetened beverages (SB) among Uruguayan children under 4, considering food security and educational context. Additionally, the intake of other foods was analyzed. Data used in this cross-sectional study were obtained in the First National Survey of Child Health, Nutrition and Development of Uruguay (n=3,077). Adult participants answered a qualitative survey. Crude regression models were tested, and adjusted for maternal age, maternal body mass index, food security and school attendance; p-value 0.046). In children ≥24 months, an inverse association was observed between school attendance and SB (p= 0.002). Those ≥24 months in the less healthy category (SB yes +0-4 healthy foods), never received breastfeeding (p= 0.001); and in the healthiest one (SB no + 4-8 healthy foods), 72.4% had been breastfeeding for 6 months or more (p= 0.001). In conclusion, food insecurity could explain ultra-processed foods and sweetened beverages consumption.

Development of a Fast Liquid Chromatography Coupled to Mass Spectrometry Method (LC-MS/MS) to Determine Fourteen Lipophilic Shellfish Toxins Based on Fused-Core Technology: In-House Validation.

Prevalence and incidence of the marine toxins (paralytic, amnesic, and lipophilic toxins) including the so-called emerging toxins (these are, gymnodimines, pinnatoxins, or spirolides among others) have increased in recent years all over the world. Climate change, which is affecting the distribution of their producing phytoplankton species, is probably one of the main causes. Early detection of the toxins present in a particular area, and linking the toxins to their causative phytoplankton species are key tools to minimize the risk they pose for human consumers. The development of both types of studies requires fast and highly sensitive analytical methods. In the present work, we have developed a highly sensitive liquid chromatography-mass spectrometry methodology (LC-MS/MS), using a column with fused-core particle technology, for the determination of fourteen lipophilic toxins in a single run of 3.6 min. The performance of the method was evaluated for specificity, linearity, precision (repeatability and reproducibility) and accuracy by analysing spiked and naturally contaminated samples. The in-house validation was successful, and the limit of detection (LOD) and quantification (LOQ) for all the toxins were far below their regulatory action limits. The method is suitable to be considered in monitoring systems of bivalves for food control.

Characterization of the Domoic Acid Uptake Mechanism of the Mussel (Mytilus galloprovincialis) Digestive Gland.

Cultures of the mussel Mytilus galloprovincialis are frequently affected by accumulation of the amnesic shellfish poisoning toxin domoic acid (DA). This species is characterized by a fast uptake and release of the toxin. In this work, the main characteristics of the uptake mechanism have been studied by incubation of digestive gland thin slices in media with different composition and DA concentration. DA uptake seems to follow Michaelis-Menten kinetics, with a very high estimated KM (1722 µg DA mL-1) and a Vmax of 71.9 µg DA g-1 h-1, which is similar to those found for other amino acids in invertebrates. Replacement of NaCl from the incubation media by Cl-choline (Na+-free medium) did not significantly reduce the uptake, but replacement by sorbitol (Na+-free and Cl--depleted medium) did. A new experiment replacing all chlorides with their equivalent gluconates (Na+- and Cl--free medium) showed an important reduction in the uptake that should be attributed to the absence of chloride, pointing to a Na+-independent, Cl- (or anion-) dependent transporter. In media with Na+ and Cl-, neither decreasing the pH nor adding cyanide (a metabolic inhibitor) had significant effect on DA uptake, suggesting that the transport mechanism is not H+- or ATP-dependent. In a chloride depleted medium, lowering pH or adding CN increased the uptake, suggesting that other anions could, at least partially, substitute chloride.

Fibromyalgia and Depression in Women: An 1H-NMR Metabolomic Study.

Fibromyalgia is a chronic and systemic syndrome characterized by muscle, bone, and joint pain. It is a gender-specific condition with a 9:1 incidence ratio between women and men. Fibromyalgia is frequently associated with psychic disorders affecting the cognitive and emotional spheres. In the reported work, we compared 31 female fibromyalgia patients to 31 female healthy controls. They were analyzed for biochemical clinical parameters, for autoimmune markers, and were subjected to 1H-NMR metabolomics analysis. To identify a correlation between the metabolomic profile and the psychic condition, a subset of 19 fibromyalgia patients was subjected to HAM-A and HAM-D Hamilton depression tests. Multivariate statistical analysis showed the dysmetabolism of several metabolites involved in energy balance that are associated with systemic inflammatory conditions. The severity of depression worsens dysmetabolic conditions; conversely, glycine and glutamate, known for their critical role as neuromodulators, appear to be potential biomarkers of fibromyalgia and are associated with different severity depression conditions.

Prenatal and Early Postnatal Cerebral d-Aspartate Depletion Influences l-Amino Acid Pathways, Bioenergetic processes, and Developmental Brain Metabolism.

d-Amino acids were believed to occur only in bacteria and invertebrates. Today, it is well known that d-amino acids are also present in mammalian tissues in a considerable amount. In particular, high levels of free d-serine (d-Ser) and d-aspartate (d-Asp) are found in the brain. While the functions of d-Ser are well known, many questions remain unanswered regarding the role of d-Asp in the central nervous system. d-Asp is very abundant at the embryonic stage, while it strongly decreases after birth because of the expression of d-aspartate oxidase (Ddo) enzyme, which catalyzes the oxidation of this d-amino acid into oxaloacetate, ammonium, and hydrogen peroxide. Pharmacologically, d-Asp acts as an endogenous agonist of N-methyl d-aspartate and mGlu5 receptors, which are known to control fundamental brain processes, including brain development, synaptic plasticity, and cognition. In this work, we studied a recently generated knockin mouse model (R26ddo/ddo), which was designed to express DDO beginning at the zygotic stage. This strategy enables d-Asp to be almost eliminated in both prenatal and postnatal lives. To understand which biochemical pathways are affected by depletion of d-Asp, in this study, we carried out a metabolomic and lipidomic study of ddo knockin brains at different stages of embryonic and postnatal development, combining nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS) techniques. Our study shows that d-Asp deficiency in the brain influences amino acid pathways such as threonine, glycine, alanine, valine, and glutamate. Interestingly, d-Asp is also correlated with metabolites involved in brain development and functions such as choline, creatine, phosphocholine (PCho), glycerophosphocholine (GPCho), sphingolipids, and glycerophospholipids, as well as metabolites involved in brain energy metabolism, such as GPCho, glucose, and lactate.