Enhancing chlamydospore production in Duddingtonia flagrans on solid substrate: The impact of mannitol and varied cultivation conditions.

Duddingtonia flagrans is a nematophagous fungus which has shown promising results as a non-chemical parasitic control tool. The fungus disrupts the parasite's life cycle by trapping larvae in the environment through the networks generated from chlamydospores, thus preventing the reinfection of animals. One barrier to the development of a commercial product using this tool is the need to increase chlamydospore production in the laboratory for its administration to livestock. The purpose of this study was to evaluate the addition of mannitol to an enriched culture medium and the effect of adverse cultivation conditions on chlamydospore production. D. flagrans was cultivated on Petri dishes with corn agar for 4 weeks at 27 °C and 70% relative humidity (RH). Four groups were then formed, all with Sabouraud agar as a base, to which different growth inducers were added: GSA (glucose Sabouraud agar), GSA-MI (glucose Sabouraud agar + meso inositol), GSA-E (enriched glucose Sabouraud agar), and AE-M (enriched agar + mannitol). After 4 weeks, chlamydospores were recovered by washing the surface of each plate with distilled water and then quantified. The medium that yielded the highest amount of chlamydospores was subjected to different cultivation conditions: NC (normal conditions): 70% RH and 27 °C, AC (adverse conditions) 1: 20% RH and 40 °C, CA2: 60% RH and 27 °C, and CA3: 55% RH and 24 °C. It was determined that mannitol increases chlamydospore production (65x106 chlamydospores/plate), and when reducing humidity by 10% under cultivation conditions it resulted in an approximately 10% increase in chlamydospore production compared to the control group. These results suggest that the addition of polyols, as well as its cultivation under certain environmental conditions, can improve chlamydospore production on a laboratory scale.

Prevalence and clinical-epidemiologic characteristics of a Mexican population with metabolic (dysfunction) associated fatty liver disease: An open population study.

INTRODUCTION AND AIMS: The risk factors for fatty liver disease are frequent in the Mexican population and the available studies appear to underestimate its prevalence. Our aims were to know the prevalence of metabolic (dysfunction) associated fatty liver disease (MAFLD) in an open population and determine the clinical and sociodemographic characteristics, nutritional status, physical activity level, and prevalence of metabolic syndrome, in the affected population. MATERIALS AND METHODS: Volunteers from the general public were invited to take part in the study. Three separate questionnaires were applied to 1) determine the sociodemographic characteristics and health status of the participants, 2) evaluate the quality of their diet, and 3) to evaluate their level of physical activity. The participants underwent somatometry, laboratory tests, liver ultrasound, and FIB-4 index determination, and transition elastography (FibroScan®) was carried out on all subjects with signs suggestive of liver fibrosis that agreed to undergo the procedure. The statistical analysis was carried out using SPSS for Windows, version 22, software. RESULTS: A total of 585 volunteers were studied, resulting in a prevalence of MAFLD of 41.3%, a predominance of men above 50 years of age, poor dietary habits, and sedentary lifestyle. Male sex, obesity, metabolic syndrome, and elevated ALT were risk factors for the disease and 40% of those affected had advanced fibrosis. CONCLUSIONS: The prevalence of MAFLD in our population is one of the highest in the world. The conditioning factors of the disease can be modified. Therefore, public policies that redirect the current trend are required.

Effect of macrocyclic lactones on nontarget coprophilic organisms: a review.

Macrocyclic lactones are frequently used dewormers in livestock farms around the world. Due to their wide spectrum of action against nematodes and arthropods and their practicality of application at very low doses, their use has become massive since their discovery. These compounds are eliminated in a large percentage in the feces of animals, causing adverse effects on coprophilic fauna. Several research groups around the world have been devoted to evaluating these effects on this fauna. The aim of this review is to register the adverse effects of the concentrations in which macrocyclic lactones are eliminated in the feces of domestic animals and the importance of the coprophilic and edaphilous fauna on the degradation of the feces of the animals. The documented data shows that the use of macrocyclic lactones has a high toxicological risk for the different species that colonize the dung, thus causing an adverse effect on its disintegration and its subsequent incorporation into the soil. Even so, more studies at the regional level and their standardization are necessary to make the comparison between different areas possible.

Biochemical characterization of the venom from the Mexican scorpion Centruroides ornatus, a dangerous species to humans.

Every year in Mexico, around 300,000 people suffer from accidents related to scorpion stings. Among the scorpion species dangerous to human is Centruroides ornatus, whose venom characterization is described here. From this venom, a total of 114 components were found using chromatographic separation and mass spectrometry analysis. The most abundant ones have molecular masses between 3000-4000 Da and 6000-8000 Da respectively, similar to other known K+ and Na+-channel specific scorpion peptides. Using intraperitoneal injections into CD1 mice, we were able to identify and fully sequenced three new lethal toxins. We propose to name them Co1, Co2 and Co3 toxins, which correspond to toxins 1 to 3 of the abbreviated species name (Co). Electrophysiology analysis of these peptides using heterologously expressed human Na+-channels revealed a typical ß-toxin effect. Peptide Co52 (the most abundant peptide in the venom) showed no activity in our in vivo and in vitro model assays. A phylogenetic analysis groups the Co1, Co2 and Co3 among other ß-toxins from Centruroides scorpions. Peptide Co52 segregates among peptides of unknown defined functions.

The S1P mimetic fingolimod phosphate regulates mitochondrial oxidative stress in neuronal cells.

Fingolimod is one of the few oral drugs available for the treatment of multiple sclerosis (MS), a chronic, inflammatory, demyelinating and neurodegenerative disease. The mechanism of action proposed for this drug is based in the phosphorylation of the molecule to produce its active metabolite fingolimod phosphate (FP) which, in turns, through its interaction with S1P receptors, triggers the functional sequestration of T lymphocytes in lymphoid nodes. On the other hand, part if not most of the damage produced in MS and other neurological disorders seem to be mediated by reactive oxygen species (ROS), and mitochondria is one of the main sources of ROS. In the present work, we have evaluated the anti-oxidant profile of FP in a model of mitochondrial oxidative damage induced by menadione (Vitk3) on neuronal cultures. We provide evidence that incubation of neuronal cells with FP alleviates the Vitk3-induced toxicity, due to a decrease in mitochondrial ROS production. It also decreases regulated cell death triggered by imbalance in oxidative stress (restore values of advanced oxidation protein products and total thiol levels). Also restores mitochondrial function (cytochrome c oxidase activity, mitochondrial membrane potential and oxygen consumption rate) and morphology. Furthermore, increases the expression and activity of protective factors (increases Nrf2, HO1 and Trx2 expression and GST and NQO1 activity), being some of these effects modulated by its interaction with the S1P receptor. FP seems to increase mitochondrial stability and restore mitochondrial dynamics under conditions of oxidative stress, making this drug a potential candidate for the treatment of neurodegenerative diseases other than MS.

Microscopio como Lector de Absorbancia con Utilidad en Análisis Clínicos / Microscope as Absorbance Reader with Utility in Clinical Analysis

Resumen La descentralización de laboratorios clínicos, se encuentra en desarrollo, lo anterior ha llevado a diseñar instrumentos que ofrecen resultados rápidos, confiables y al lado del paciente, esta tendencia se le conoce como prueba en el punto de atención (point of core testing POCT) y brinda la posibilidad de dar un seguimiento inmediato al padecimiento. El objetivo de esta investigación fue la implementación de un medidor de absorbancia, empleando la estructura de un microscopio óptico, al cual se le ha adaptado un filtro de luz, y una cámara digital. Lo anterior permite obtener valores de intensidad promedio de imágenes sólidas microscópicas de reacciones enzimáticas, y a partir de ellas estimar absorbancia o concentración. Como resultados se presentan rectas de calibración de absorbancia y mediciones de concentraciones para los casos de violeta de genciana, un kit de glucosa oxidasa y muestras problemas de pacientes voluntarios. Concluimos que existe un error de medición menor de ±1mg/dL comparados con las mediciones de un lector de placas de Elisa y un analizador de química sanguínea semiautomatizado. Teniendo en cuenta lo anterior el sistema resulta ser una alternativa viable para la estimación de absorbancia y aumenta la funcionalidad de microscopios ópticos en clínicas de salud.

Abstract The decentralization of clinical laboratories is under development, which has led to the design of instruments that offer fast, reliable and patient-side results, this trend is known as point of core testing (POCT) and It offers the possibility of giving an immediate follow-up to the disease. The objective of this investigation was the implementation of an absorbance meter, using the structure of an optical microscope, to which a light filter and a digital camera have been adapted. This allows to obtain values of average intensity of solid images of enzymatic reactions, and from them to estimate absorbance or concentration. As results, we present absorbance calibration lines and concentration measurements for cases of gentian violet, a glucose oxidase kit and samples of volunteer patients. We conclude that there is a measurement error of less than ± 1mg / dl compared with the measurements of an Elisa plate reader and a semi-automated blood chemistry analyzer. Taking into account the above, the system turns out to be a viable alternative for estimating absorbance and increasing the functionality of optical microscopes in health clinics.

Cytotoxic T lymphocyte-associated antigen 4 rs231775 polymorphism and osteosarcoma.

Systematic reviews on osteosarcoma have concluded that CTLA4 rs231775 AA genotype influences risk for the disease in the Chinese population. Remarkably, rs231775 shows different frequencies in different human populations. Therefore, it would be interesting to know whether this SNP is related to the risk of osteosarcoma in other populations. The present study aimed to evaluate the association between rs231775 and the susceptibility of osteosarcoma in the Spanish population. We performed an updated meta-analysis including a total of 538 cases and 623 controls. The genotypic association analyses showed that the CTLA4 rs231755 was associated with osteosarcoma susceptibility in the Spanish population. When meta-analysis was performed, the results displayed that CTLA4 rs231775 AA genotype was associated with the risk of developing osteosarcoma in all analyzed populations (OR=2.07; 95% CI: 1.48-2.89).The rs231775 AA genotype could be considered as a susceptibility marker in osteosarcoma Keywords: CTLA4, rs231775, +49A/G and osteogenic sarcoma.

Gene therapy with mesenchymal stem cells expressing IFN-ß ameliorates neuroinflammation in experimental models of multiple sclerosis.

BACKGROUND AND PURPOSE: Recombinant IFN-ß is one of the first-line treatments in multiple sclerosis (MS), despite its lack of efficacy in some patients. In this context, mesenchymal stem cells (MSCs) represent a promising therapeutic alternative due to their immunomodulatory properties and multipotency. Moreover, by taking advantage of their pathotropism, these cells can be genetically modified to be used as carriers for delivering or secreting therapeutic drugs into injured tissues. Here, we report the therapeutic effect of systemic delivery of adipose-derived MSCs (AdMSCs), transduced with the IFN-ß gene, into mice with experimental autoimmune encephalomyelitis (EAE). EXPERIMENTAL APPROACH: Relapsing-remitting and chronic progressive EAE were induced in mice. Cells were injected i.v. Disease severity, inflammation and tissue damage were assessed clinically, by flow cytometry of spleens and histopathological evaluation of the CNS respectively. KEY RESULTS: Genetic engineering did not modify the biological characteristics of these AdMSCs (morphology, growth rate, immunophenotype and multipotency). Furthermore, the transduction of IFN-ß to AdMSCs maintained and, in some cases, enhanced the functional properties of AdMSCs by ameliorating the symptoms of MS in EAE models and by decreasing indications of peripheral and central neuro-inflammation. CONCLUSION AND IMPLICATIONS: Gene therapy was found to be more effective than cell therapy in ameliorating several clinical parameters in both EAE models, presumably due to the continuous expression of IFN-ß. Furthermore, it has significant advantages over AdMSC therapy, and also over systemic IFN-ß treatment, by providing long-term expression of the cytokine at therapeutic concentrations and reducing the frequency of injections, while minimizing dose-limiting side effects.

Antoni de Gimbernat and Barcelona (1760 - 1779)

Barcelona is a most important city in the life of Antonio Gimbernat Arbós. In the period in which he was living there, 1760 to 1779, he completed his training, he initiated and developed his teaching as professor of the Royal College of Surgeons, he conducted research of high quality describing anatomical structures of the femoral ring, and he described his new surgical technique for hernia, which practiced as major surgeon of the Hospital de la Santa Creu. He also founded his family, married and had six children, all born in Barcelona. At this period, he carried out a European tour for the acquisition of new knowledge that he first applied in Barcelona, and later in the capital of the Spanish kingdom, in which he achieved the top in management and health accountability of his time, as well as great fame (AU)

No disponible

A transcriptomic analysis of the effect of genistein on Sinorhizobium fredii HH103 reveals novel rhizobial genes putatively involved in symbiosis.

Sinorhizobium fredii HH103 is a rhizobial soybean symbiont that exhibits an extremely broad host-range. Flavonoids exuded by legume roots induce the expression of rhizobial symbiotic genes and activate the bacterial protein NodD, which binds to regulatory DNA sequences called nod boxes (NB). NB drive the expression of genes involved in the production of molecular signals (Nod factors) as well as the transcription of ttsI, whose encoded product binds to tts boxes (TB), inducing the secretion of proteins (effectors) through the type 3 secretion system (T3SS). In this work, a S. fredii HH103 global gene expression analysis in the presence of the flavonoid genistein was carried out, revealing a complex regulatory network. Three groups of genes differentially expressed were identified: i) genes controlled by NB, ii) genes regulated by TB, and iii) genes not preceded by a NB or a TB. Interestingly, we have found differentially expressed genes not previously studied in rhizobia, being some of them not related to Nod factors or the T3SS. Future characterization of these putative symbiotic-related genes could shed light on the understanding of the complex molecular dialogue established between rhizobia and legumes.

Recombinant soluble IFN receptor (sIFNAR2) exhibits intrinsic therapeutic efficacy in a murine model of Multiple Sclerosis.

Endogenous interferon beta (IFNß) is an important cytokine involved in several chronic inflammatory diseases, such as Multiple Sclerosis (MS). In spite of the numerous therapeutic approaches available for MS patients, the administration of recombinant IFNß continues being one of the first line treatment to these patients. The soluble form of IFNß receptor (sIFNAR2) could act as critical regulator of the endogenous and the systemically administered IFNß, but whether it functions as an agonist or antagonist of its ligand is not completely elucidated. Morover, the possible role of sIFNAR2 in autoimmune diseases like MS is still unknown and so far overlooked. Here we evaluated the efficacy of the combined therapy of IFNß and our recombinant protein analogous to human sIFNAR2 as a treatment in a chronic mice model of MS (CP-EAE). We also tested the effect of the sIFNAR2 administered as a monotherapy over these EAE-animals. The results showed that our recombinant sIFNAR2 protein potentiates the immunomodulatory effects of exogenous IFNß in CP-EAE by increasing the reduction of the induced inflammation and the tissue damage. Furthermore, we demonstrate for the first time that sIFNAR2 shows intrinsic properties by modulating the CP-EAE progression and the neuroinflammation processes related to this disease. Another intrinsic activity showed by sIFNAR2 is the inhibition of the T cells proliferation, which increase its potential as therapeutic molecule.

Overexpression of connexin 43 reduces melanoma proliferative and metastatic capacity.

BACKGROUND: Alterations in connexin 43 (Cx43) expression and/or gap junction (GJ)-mediated intercellular communication are implicated in cancer pathogenesis. Herein, we have investigated the role of Cx43 in melanoma cell proliferation and apoptosis sensitivity in vitro, as well as metastatic capability and tumour growth in vivo. METHODS: Connexin 43 expression levels, GJ coupling and proliferation rates were analysed in four different human melanoma cell lines. Furthermore, tumour growth and lung metastasis of high compared with low Cx43-expressing FMS cells were evaluated in vivo using a melanoma xenograft model. RESULTS: Specific inhibition of Cx43 channel activity accelerated melanoma cell proliferation, whereas overexpression of Cx43 increased GJ coupling and reduced cell growth. Moreover, Cx43 overexpression in FMS cells increased basal and tumour necrosis factor-α-induced apoptosis and resulted in decreased melanoma tumour growth and lower number and size of metastatic foci in vivo. CONCLUSIONS: Our findings reveal an important role for Cx43 in intrinsically controlling melanoma growth, death and metastasis, and emphasise the potential use of compounds that selectively enhance Cx43 expression on melanoma in the future chemotherapy and/or immunotherapy protocols.

[Antifungal susceptibility of clinical isolates of Scopulariopsis species]. / Sensibilidad a antifúngicos de especies de Scopulariopsis de origen clínico.

INTRODUCTION: Scopulariopsis is a common soil saprophyte. In the last years the infections caused by Scopulariopsis species have increased, included superficial and invasive mycoses. This fungi has been reported resistant in vitro to some antifungal agents, although there is little information about this. The aim of the study was to establish in vitro antifungal susceptibility of clinical isolates of Scopulariopsis species against to broad-spectrum antifungal agents. METHODS: A total of 28 Scopulariopsis strains (10 S. brevicaulis, 7 S. koningii, 3 S. acremonium, 3 S. candida, 3 S. flava, 1 S. brumptii and 1 S. fusca) were tested using Sensititre Yeast One and broth microdilution methods to determine the minimum inhibitory concentrations (MICs) to amphotericin B, fluconazole, itraconazole, posaconazole, voriconazole and 5-fluorocytosine, and minimun effective concentration (MECs) to anidulafungin, caspofungin and micafungin. RESULTS: Our data confirm the high in vitro resistance of Scopulariopsis to antifungal agents. Anidulafungin, caspofungin, micafungin (MICs ≥ 8 mg/L), 5-fluorocytosine (MICs ≥ 64 mg/L), and fluconazole (MICs ≥ 128 mg/L) were inactive in vitro in all species. MICs of amphotericin B (range 2 to ≥ 8 mg/L) and itraconazole (0.5 to ≥ 16 mg/L) were high. The best antifungal activity was observed for posaconazole and voriconazole (0.5 to ≥ 8 mg/L). With Sensititre Yeast One method MICs obtained slightly lower. Scopulariopsis candida, S. flava and S. fusca were the most resistant species, while S. acremonium and S. brevicaulis showed the lowest MICs. CONCLUSIONS: MICs of all tested antifungal agents for Scopulariopsis were very high. Infections caused by Scopulariopsis species may not respond to antifungal treatment. Voriconazole is the drug of choice for treatment. We consider it appropriate to add amphotericin B in serious infections.

Plant growth promotion in cereal and leguminous agricultural important plants: from microorganism capacities to crop production.

Plant growth-promoting rhizobacteria (PGPR) are free-living bacteria which actively colonize plant roots, exerting beneficial effects on plant development. The PGPR may (i) promote the plant growth either by using their own metabolism (solubilizing phosphates, producing hormones or fixing nitrogen) or directly affecting the plant metabolism (increasing the uptake of water and minerals), enhancing root development, increasing the enzymatic activity of the plant or "helping" other beneficial microorganisms to enhance their action on the plants; (ii) or may promote the plant growth by suppressing plant pathogens. These abilities are of great agriculture importance in terms of improving soil fertility and crop yield, thus reducing the negative impact of chemical fertilizers on the environment. The progress in the last decade in using PGPR in a variety of plants (maize, rice, wheat, soybean and bean) along with their mechanism of action are summarized and discussed here.

Una tesis pionera en la Medicina de Urgencias y Emergencias española: “Alcance de la cirujía [sic]de urgencias en las casas de socorro” (1907) / Pioneer 1907 doctoral thesis on emergency medicine in Spain: “The scope of emergency

Este artículo descubre, comenta y revisa la tesis doctoral defendida por el doctor Manuel Pascual Alonso en 1907. En ella se expone el estado material e instrumental, agentes médicos implicados y las prácticas quirúrgicas de alcance en las casas de socorro, verdaderos antecedentes de los servicios médicos de urgencia actuales. Tal tesispionera es un hallazgo relevante e iluminativo para la medicina española de urgencias y emergencias y que pone de manifiesto, a partir de este estudio retrospectivo, una valiosa y multisecular tradición médica de urgencias en España desconsiderada hasta ahora (AU)

This review and commentary of the doctoral thesis defended by Dr. Manuel Pascual Alonso in 1907, summarizes this pioneering author’s account of the material and instrumental conditions in the emergency care facilities known as casas de socorro (first-aid or rescue facilities), the true forerunners of today’s emergency health services. Dr. Pascual describes physician staffing and surgical practices in these centers. The thesis sheds light on the history of Spanish emergency medicine, revealing that Spain has a worthy, centuries-long tradition of emergency medical practice that has been underappreciated until now (AU)

Aneurisma de tronco tibioperoneo. A propósito de un caso / Tibioperoneal trunk aneurysm. A case report

No disponible