Liquid Chromatography-Mass Spectrometry-Based Metabolomics for Understanding the Compositional Changes Induced by Oxidative or Anoxic Storage of Red Wines.

The aim of this work was to study the physicochemical changes of eight red wines stored under conditions differing in O2 exposure and temperature and time under anoxia. The methods used to analyze the wines included the measurement of volatile sulfur compounds, color, tannin (T) polymerization, and liquid chromatography-mass spectrometry untargeted metabolomic fingerprint. After 3 months, the color of the oxidized samples evolved 4-5 times more intensively than in wines stored under anoxia. The major metabolomic differences between oxidative and anoxic conditions were linked to reactions of acetaldehyde (favored in oxidative) and SO2 (favored in anoxia). In the presence of oxygen, the C-4 carbocation of flavanols delivered ethyl-linked tannin-anthocyanin (T-A) and tannin-tannin (T-T) adducts, pyranoanthocyanins, and sulfonated indoles, while under reduction, the C-4 carbocation delivered direct linked T-A adducts, rearranged T-T adducts, and sulfonated tannins. Some of these last reactions could be related to the accumulation of reduced species, eventually ending with reductive off-odors.

Comparison of PCR-based methods for the diagnosis of cutaneous leishmaniasis in two different epidemiological scenarios: Spain and Morocco.

BACKGROUND: Cutaneous leishmaniasis (CL) is a disfiguring and stigmatising disease occurring in more than 70 countries across the world including Spain and Morocco. The use of sensitive tests that can differentiate Leishmania species is advised. OBJECTIVE: To evaluate the influence of the epidemiological scenario on the reliability of the PCR techniques and contribute to the selection of the most efficient one for CL diagnosis. METHODS: The sensitivities of parasitological methods and four PCRs were compared in cutaneous samples from 77 patients from Spanish (PSH) and Moroccan hospitals (PMH). Exudates and fresh or paraffin-embedded tissue biopsies were used. RESULTS: None of the PCRs used in this study allowed the diagnosis of all CL cases, showing also some drawbacks. Lmj4/Uni21-PCR displayed the best sensitivity with PMH, but it did not provide positive results in PSH with CL confirmed by other PCRs. Conversely, JW13/JW14-PCR and L. infantum-PCR-ELISA displayed good sensitivities with PSH that were not achieved with PMH. Nested-ITS-1-PCR did not show enough sensitivity with paraffin-embedded tissue biopsies. False-negative results were obtained in 19% of PSH due to unspecific hybridizations of ITS-1 primers with human chromosome1. CONCLUSIONS: PCR should be routinely used in patients with cutaneous lesions compatible with CL and furthermore, the combination of two PCR techniques is advisable. The selection of these PCRs will be influenced by the epidemiological scenario: In areas where L. infantum is endemic, the use of the PCR-ELISA joint with JW13/JW14-PCR seems an appropriate choice, whereas in areas such as Morocco, Lmj4/Uni21 and ITS-1 provide satisfactory results.

Lipid nanoparticles (SLN & NLC) for delivery of vitamin E: a comprehensive review.

The antioxidative and photoprotective properties of vitamin E have caused it to be included as an active agent in various pharmaceutical and cosmetic products. However, its lipophilicity, chemical instability and poor skin penetration have limited the effectiveness of these formulations. For that reason, many attempts to include it in different drug delivery systems have been made. In recent decades, lipid nanoparticles have received special attention due to their advantages of compatibility with the skin, ability to enhance penetration of drugs in the stratum corneum, protection of the encapsulated substance against degradation induced by the external medium and control of drug release. This work reviews the current status of the encapsulation of vitamin E in lipid nanoparticles. We describe the most important methods for obtaining and characterizing lipid nanoparticles containing vitamin E (LNP-VE), various techniques for the evaluation of vitamin E's properties after encapsulation, the main in vitro and in vivo studies of the potential effectiveness or toxicity of LNP-VE, the formulations and stability studies of this delivery system, the commercial products based on LNP-VE and the regulatory aspects related to lipid nanoparticles. Finally, we discuss the most relevant advantages of encapsulating vitamin E in such particles and critical aspects that still demand attention to enhance the potential of solid lipid nanoparticles to deliver vitamin E.

Effectiveness of the sesquiterpene (-)-α-bisabolol in dogs with naturally acquired canine leishmaniosis: an exploratory clinical trial.

The use of natural products is a promising approach for treating visceral leishmaniosis. (-)-α-Bisabolol is a sesquiterpene that have been proved active in vivo on Leishmania infantum-infected mice without showing toxicity. A single-centre, parallel-group, randomized, exploratory study was designed to assess its efficacy in a canine leishmaniosis model involving naturally infected dogs. In this clinical trial, 12 dogs were allocated into two groups and were treated with either meglumine antimoniate (100 mg/kg) through subcutaneous route or (-)-α-bisabolol (30 mg/kg) through oral route for two treatment series of 30 days, separated by a 30-day interval. A 4-month follow-up period was established as well. Parasite loads in bone marrow, lymph node and blood were estimated through quantitative PCR. Antibody titres were determined through immunofluorescence antibody test and cytokine expression values were estimated through real-time reverse transcription-PCR. Treatment safety was assessed through the evaluation of weight, gastrointestinal alterations and hematological and biochemical parameters in blood. Analyses were performed before and after treatment, and after a 4-months follow-up period. Treatment with the sesquiterpene was effective at decreasing parasite loads and increasing gamma-interferon expression level. Dogs treated with (-)-α-bisabolol did not show any toxicity sign. These results were better than those obtained using the reference drug, meglumine antimoniate. The natural compound seemed to induce a Th1 immune response that led to parasitological and clinical improvement without showing any safety issue, suggesting a high potential for the treatment of canine and human visceral leishmaniosis.

Cutaneous leishmaniasis by Leishmania infantum: behind granulomatous lesions of unknown aetiology.

BACKGROUND: Cutaneous leishmaniasis (CL) is underestimated in Spain as in other European countries due to the polymorphism of its clinical manifestations and histopathological features discouraging doctors from suspecting leishmaniasis. Mucosal manifestations (ML) are misdiagnosed due to the fact that they often mimic cancer. OBJECTIVES: Given that leishmaniasis may be masked as different granulomatous diseases in Leishmania infantum endemic areas, the aim of this study was to verify this misdiagnosing and contributes to the improvement of CL/ML diagnosis. METHODS: A retrospective study involving formalin-fixed paraffin-embedded tissue biopsies with histopathological features of granulomatous lesions of unknown origin (GLUO) detected in 17 patients. This study included 13 patients with CL that was used as positive controls, nine patients with other confirmed diseases used as negative controls and seven patients with histological features suggestive of CL or ML without confirmation. Molecular analysis was blindly performed using two different PCR techniques. RESULTS: The PCR detected 15 CL cases in which the diagnosis was neither clinically nor histologically suspected. Leishmaniasis was confirmed in seven suspected patients in whom the classical techniques failed to detect the parasite. L. infantum was identified in all cases. A systematic review of CL cases in GLUO patients from European countries identified 45 reported cases. CONCLUSIONS: In L. infantum endemic areas, a high percentage of GLUO are due to Leishmania infection. The main consequences are delayed diagnosis and underestimation of the real incidence. PCR performed on paraffin-embedded tissue proved to be a reliable tool for diagnosis of CL/ML and must be performed routinely in any granulomatous dermatitis, even when the morphological features are no stereotypical of leishmaniasis.

Encefalopatía hepática: el manganeso también brilla en T1 / Hepatic encephalopathy: manganese also shines in T1

No disponible

Turning on the red phosphorescence of a [Ru(tpy)(bpy)(Cl)]Cl complex by amide substitution: self-aggregation, toxicity, and cellular localization of an emissive ruthenium-based amphiphile.

Coupling the notoriously non-emissive complex [Ru(tpy)(bpy)Cl]Cl (tpy = 2,2':6',2''-terpyridine, bpy = 2,2'-bipyridine) to a C12 alkyl chain via an amide linker on the 4' position of the terpyridine yielded a new amphiphilic ruthenium complex showing red emission and chloride-dependent aggregation properties. This emissive complex is highly cytotoxic in A549 non-small lung cancer cells where it can be followed by confocal microscopy. Uptake occurs within minutes, first by insertion into the cellular membrane, and then by migration to the peri-nuclear region.

The sesquiterpene (-)-α-bisabolol is active against the causative agents of Old World cutaneous leishmaniasis through the induction of mitochondrial-dependent apoptosis.

Cutaneous leishmaniasis treatment remains challenging due to the absence of a satisfactory treatment. The screening of natural compounds is a valuable strategy in the search of new drugs against leishmaniasis. The sesquiterpene (-)-α-bisabolol is effective in vivo against visceral leishmaniasis due to Leishmania infantum, but its mechanism of action remains elusive. The aim of this study is to validate this promising compound against the causative species of Old World cutaneous leishmaniasis and to get an insight into its antileishmanial mode of action. The compound was evaluated on L. tropica promastigotes and intracellular amastigotes using bone marrow-derived macrophages and its cytotoxicity was evaluated on L929 fibroblasts. The reactive oxygen species generation was evaluated using a sensitive probe. Mitochondrial depolarization was assessed evaluating the fluorescence due to rhodamine 123 in a flow cytometer. Apoptosis was investigated by measuring the fluorescence due to annexin V and propidium iodide in a flow cytometer. The ultrastructure of treated promastigotes and intracellular amastigotes was analysed through transmission electron microscopy. (-)-α-Bisabolol was active against L. tropica intracellular amastigotes displaying an inhibitory concentration 50 % of 25.2 µM and showing low cytotoxicity. This compound induced time and dose-dependent oxidative stress, mitochondrial depolarization and phosphatidilserine externalization (a marker of apoptosis). These effects were noticed at a low concentration and short exposure time. In the ultrastructural analyses, the treated parasites showed mitochondrial disruption, presence of electron-dense structures and chromatin condensation. These results suggest that this natural compound induces oxidative stress and mitochondrial-dependent apoptosis on Leishmania without disturbing the plasma membrane.

Genetic variability and infective ability of the rabbit trypanosome, Trypanosoma nabiasi Railliet 1895, in southern Spain.

Trypanosomes are widespread haemoflagellate protozoans, commonly found in all groups of vertebrates and usually transmitted by arthropods. Non-pathogenic species are those that cause little or no apparent negative effects in the host and it is accepted that Trypanosoma nabiasi is the species that infects the domestic and wild rabbit, Oryctolagus cuniculus. Knowledge about genetic variability, in vitro cultivation and infectivity of this parasite is very scarce, so the aim of this study was to provide an insight on them. The parasite was detected in all the type of samples of 121 wild rabbits. Epimastigotes were visualized and isolated from all the organ cultures types except from skin, and twenty-six strains were isolated and grown in mass. Epimastigote infectivity was assessed in vitro and in vivo. Amastigotes were obtained in infected macrophages from cultured epimastigotes. Furthermore, trypomastigotes were found in the peripheral bloodstream of an experimentally infected naïve domestic rabbit with cultured epimastigotes at the fourth day after infection. The rising titre of antibodies led to the disappearance of the parasite from blood. In addition, this study reports the existence of two T. nabiasi genetic lineages in southern Spain. Phylogenetic analysis places T. nabiasi in the same clade as T. lewisi and other rodent trypanosomes of the subgenus Herpetosoma.

Hair parasite load as a new biomarker for monitoring treatment response in canine leishmaniasis.

Canine leishmaniasis treatment focuses on the reduction of parasite load, the clinical improvement of the animal, and the avoidance of relapses, in a scenario where the definitive parasite clearance is not achievable. Therefore, monitoring is crucial during the treatment of this disease. Quantitative PCR has been shown as an ideal tool for the treatment monitoring when quantifying parasite load in target organs such as lymph node or bone marrow, tissues that are too invasive for regular evaluation. This study aims to prove the potential of hair parasite load in the treatment monitoring of canine leishmaniasis. Six dogs were treated with meglumine antimoniate and monitored up to four months after the end of the treatment. Parasite loads in bone marrow, blood, lymph node and hair were quantified by real-time quantitative PCR. Total IgG, IgG1, and IgG2 antibody titres were analysed by immunofluorescent assay and a clinical assessment was carried out. Treatment consisted of two 28-day courses of meglumine antimoniate (100mg/kg/day) separated by an one-month interval. Analyses were performed before (day 0), during (day 60) and after treatment (day 120), and at the end of a follow-up period (day 210, four months after the end of treatment). Hair parasite load turned out to be strongly correlated with bone marrow, lymph node and blood parasite loads and with the clinical score and the IgG1 antibody titre. The evolution of this biomarker reflects the evolution of the parasitological, immunological and clinical state of the dog, highlighting its potential as a non-invasive marker for the treatment monitoring in canine leishmaniasis.

Accesibilidad al sistema sanitario de los pacientes diabéticos con síndrome coronario agudo con elevación del segmento ST / Accessibility to health care of diabetic patients with acute coronary syndrome ST-segment elevation

OBJETIVOS: El objetivo de este estudio es medir la accesibilidad al sistema sanitario de los pacientes diabéticos y analizar si las posibles diferencias en la accesibilidad explican la mayor mortalidad conocida en aquellos. MÉTODOS: Estudio de cohortes retrospectivo, realizado en pacientes diabéticos con síndrome coronario agudo con elevación del segmento ST incluidos en los años 2010 al 2013 del registro ARIAM-SEMICYUC. Se realiza análisis crudo y ajustado mediante regresión logística no condicional. RESULTADOS: Se han analizado 4817 pacientes, de los cuales 1070 (22,2%) son diabéticos. Los pacientes diabéticos contactan con el sistema sanitario de la misma forma que los pacientes no diabéticos aunque con mayor retraso (retraso atribuible al paciente 90 min vs. 75 min con p = 0,004 y retraso prehospitalario 150 min vs. 130 min con p = 0,002). Una vez dentro del sistema sanitario, estos pacientes tienen menor tasa de reperfusión (50 vs. 57,7%; p < 0,001) pero sin objetivar mayor retraso en el tratamiento. Como ya es conocido, los pacientes diabéticos presentan una mayor mortalidad hospitalaria (12,5 vs. 6%; p < 0,001); sin embargo, no se identifican como variables predictoras independientes de la mortalidad ni el retraso atribuible al paciente ni el retraso prehospitalario. CONCLUSIONES: Los pacientes diabéticos tienen una mayor demora en el acceso al sistema sanitario, sin embargo no hemos podido objetivar que esta demora se relacione de forma independiente con la mayor mortalidad

OBJECTIVES: To measure accessibility to health care among diabetic patients and analyze whether differences in delay explain differences in hospital mortality. METHODS: A retrospective cohort study was conducted in diabetic patients with acute coronary syndrome with ST-segment elevation included in the ARIAM-SEMICYUC registry (2010-2013). Crude and adjusted analyses were performed using unconditional logistic regression. RESULTS: A total of 4817 patients were analyzed, of whom 1070 (22.2%) were diabetics. No differences were found in access to health care between diabetic and non-diabetic patients. Diabetic patients presented with longer patient delay (90 min vs. 75 min; p = .004) and prehospital delay (150min vs. 130 min; p = .002). Once the health system was contacted, diabetic patients had a lower reperfusion rate (50% vs. 57.7%; p < .001), but no longer delay in treatment was observed compared with the non-diabetic individuals. Diabetic patients have greater in-hospital mortality (12.5 vs. 6%; p < .001), though neither patient delay nor prehospital delay were identified as independent predictors of in-hospital mortality. CONCLUSIONS: Diabetic patients had a longer delay in access to health care, though such delay was not independently related to increased mortality

Differential ecological traits of two Phlebotomus sergenti mitochondrial lineages in southwestern Europe and their epidemiological implications.

OBJECTIVES: The introduction of leishmaniasis in a new area requires a well-established population of the sandfly vector species of the parasite. No autochthonous cases of anthroponotic cutaneous leishmaniasis have been detected in southwestern Europe, and Leishmania infantum is the only causative agent of leishmaniasis in this area. Phlebotomus sergenti, the main vector of Leishmania tropica, is commonly found in the Iberian Peninsula at sufficient densities to be able to act as a vector. It is characterised by high genetic diversity and classified in four mitochondrial lineages. Our aim was to analyse the composition and distribution of P. sergenti mitochondrial lineages in southwestern Europe given the possibility of phenotypic differences of biomedical importance between them. METHODS: Sandflies were captured in the Iberian Peninsula and on the Canary and Balearic Islands. Mitochondrial lineage identification of 137 P. sergenti was performed using a novel PCR-RFLP that avoids the necessity of gene sequencing. RESULTS: Two lineages were evidenced, the typical Iberian one (lineage I) and another, held in common with North Africa (lineage III), that show a distinctive distribution. P. sergenti lineage I shows a better correlation to the bioclimatic diversity in southwestern Europe. Conversely, P. sergenti lineage III prefers warmer temperatures and less precipitation, which are typical of the Mediterranean. CONCLUSION: Lineage I seems to have adaptive advantages given its wider tolerance to temperature and altitude than lineage III, and it would seem more suitable to lead a potential geographical expansion towards the rest of Europe.

Carcinoma palpebral de células de Merkel. Serie de 5 casos y revisión de la literatura / Merkel cell carcinoma of the eyelid. A series of 5 cases and review of the literature

OBJETIVO: Estudio descriptivo de 5 casos clínicos de carcinoma de Merkel a nivel palpebral, destacando las características clínicas, histopatológicas, el manejo y seguimiento. Revisión de literatura asociada. MÉTODOS: Revisión retrospectiva de fichas clínicas y entrevista telefónica a 5 pacientes tratados por carcinoma de células de Merkel entre los años 2006-2013, en el Departamento de Órbita y Oculoplástica del Hospital Clínico de la Universidad de Chile. RESULTADOS: Cinco pacientes (2 hombres, 3 mujeres), de 79,2 años (rango 64-94 años), con una evolución de 10 semanas (rango 5-16 semanas), con un tamaño tumoral de 2,5 × 2 X 2,1 cm en promedio al momento de la cirugía. Descritos como una masa nodular, exofítica, sólida, de color rojizo. Sin adenopatías sospechosas ni metástasis en el primer enfrentamiento. Tipificados como T2N0M0, tras la primera cirugía con criterios oncológicos. Se observó, durante el seguimiento, 2 recurrencias en nódulo linfoide. Encontramos metástasis a distancia en un paciente. El diagnóstico histopatológico se confirmó mediante biopsia corriente más inmunohistoquímica. La estrategia quirúrgica fue con extirpación completa, control de márgenes libres y reconstrucción palpebral con colgajo de Hughes, Cutler-Beard o cierre primario. CONCLUSIONES: El diagnóstico precoz, la extirpación amplia del tumor con control de márgenes libres intraoperatorio con cirugía convencional o técnica de Mohs y una adecuada reconstrucción palpebral permiten una supervivencia en pacientes añosos portadores de este tumor a nivel palpebral. Se recomienda el estudio de ganglio centinela y biopsia en la intervención primaria, junto a una radioterapia posterior, que disminuye la recurrencia y aumentaría la supervivencia

OBJECTIVE: Presentation of 5 clinical cases of Merkel cell carcinoma of the eyelid, highlighting the clinical, histopathological, management, and monitoring features. Review of related literature. METHODS: Retrospective review of clinical records and telephone interview of the 5 patients treated for Merkel cell carcinoma between 2006 and 2013, in the Orbit and Oculoplastic Department, Clinical Hospital of the University of Chile. RESULTS: Five patients (2 men, 3 women); 79.2 years (range 64-94 years), with a mean onset of 10 weeks (range 5-16 weeks), tumour size reaching a mean of 2.5 × 2 × 2.1 cm at the time of surgery. Described as a nodular mass, exophytic, solid, reddish coloured. With no infiltrated lymph nodes or metastases in the first match. Staging as T2N0M0, after the first surgery with oncological criteria. Two lymph node recurrences detected during monitoring. Distant metastasis was found in one patient. The histopathological diagnosis was confirmed by immunohistochemical study of the biopsy. The surgical strategy was full excision, control of surgical margins in the intraoperative period, and eyelid reconstruction with Hughes flap, Cutler/Beard flap or primary closure, depending on the case. CONCLUSIONS: Early diagnosis, wide excision of the tumour with intraoperative control of clear margins with conventional or Mohs surgery, and proper eyelid reconstruction are adequate for a good survival in elderly patients with this eyelid tumour. The study of sentinel lymph node biopsy in primary intervention is recommended; with subsequent radiotherapy to decrease the recurrence and increase survival

Merkel cell carcinoma of the eyelid. A series of 5 cases and review of the literature.

OBJECTIVE: Presentation of 5 clinical cases of Merkel cell carcinoma of the eyelid, highlighting the clinical, histopathological, management, and monitoring features. Review of related literature. METHODS: Retrospective review of clinical records and telephone interview of the 5 patients treated for Merkel cell carcinoma between 2006 and 2013, in the Orbit and Oculoplastic Department, Clinical Hospital of the University of Chile. RESULTS: Five patients (2 men, 3 women); 79.2 years (range 64-94 years), with a mean onset of 10 weeks (range 5-16 weeks), tumour size reaching a mean of 2.5×2×2.1cm at the time of surgery. Described as a nodular mass, exophytic, solid, reddish coloured. With no infiltrated lymph nodes or metastases in the first match. Staging as T2N0M0, after the first surgery with oncological criteria. Two lymph node recurrences detected during monitoring. Distant metastasis was found in one patient. The histopathological diagnosis was confirmed by immunohistochemical study of the biopsy. The surgical strategy was full excision, control of surgical margins in the intraoperative period, and eyelid reconstruction with Hughes flap, Cutler/Beard flap or primary closure, depending on the case. CONCLUSIONS: Early diagnosis, wide excision of the tumour with intraoperative control of clear margins with conventional or Mohs surgery, and proper eyelid reconstruction are adequate for a good survival in elderly patients with this eyelid tumour. The study of sentinel lymph node biopsy in primary intervention is recommended; with subsequent radiotherapy to decrease the recurrence and increase survival.

[Accessibility to health care of diabetic patients with acute coronary syndrome ST-segment elevation]. / Accesibilidad al sistema sanitario de los pacientes diabéticos con síndrome coronario agudo con elevación del segmento ST.

OBJECTIVES: To measure accessibility to health care among diabetic patients and analyze whether differences in delay explain differences in hospital mortality. METHODS: A retrospective cohort study was conducted in diabetic patients with acute coronary syndrome with ST-segment elevation included in the ARIAM-SEMICYUC registry (2010-2013). Crude and adjusted analyses were performed using unconditional logistic regression. RESULTS: A total of 4817 patients were analyzed, of whom 1070 (22.2%) were diabetics. No differences were found in access to health care between diabetic and non-diabetic patients. Diabetic patients presented with longer patient delay (90 min vs. 75 min; p=.004) and prehospital delay (150 min vs. 130 min; p=.002). Once the health system was contacted, diabetic patients had a lower reperfusion rate (50% vs. 57.7%; p<.001), but no longer delay in treatment was observed compared with the non-diabetic individuals. Diabetic patients have greater in-hospital mortality (12.5 vs. 6%; p <.001), though neither patient delay nor prehospital delay were identified as independent predictors of in-hospital mortality. CONCLUSIONS: Diabetic patients had a longer delay in access to health care, though such delay was not independently related to increased mortality.

Neumotórax persistente en paciente politraumatizado / Persistant pneumothorax in a polytraumatized patient

No disponible