Exploring the utility of Geometric Morphometrics to analyse prehistoric hand stencils.

Hand stencils are a remarkable graphic expression in Prehistoric rock art, dating back to 42 ka BP. Although these stencils provide direct impressions of the artists' hands, the characterization of their biological profile (i.e., biological sex and age) is very challenging. Previous studies have attempted this analysis with traditional morphometrics (TM), whereas little research has been undertaken using Geometric Morphometrics (GM), a method widely used in other disciplines but only tentatively employed in rock art studies. However, the large variation in relative finger position in archaeological hands poses the question of whether these representations can be examined through GM, or, in contrast, if this creates an unmanageable error in the results. To address this issue, a 2D hand scans sample of 70 living individuals (F = 35; M = 35) has been collected in three standardized positions (n = 210) and digitized with 32 2D conventional landmarks. Results show that the intra-individual distance (mean Procrustes distance between Pos. 1-2 = 0.132; 2-3 = 0.191; 1-3 = 0.292) is larger than the inter-individual distance (mean in 1 = 0.122; 2 = 0.142; 3 = 0.165). Finally, it has been demonstrated that the relative finger positions, as well as the inclusion of all hand parts in the analysis, have an overshadowing effect on other variables potentially involved in the morphometric variability of the hand, such as biological sex.

Estudio comparativo de melatonina contra los tratamientos inmunomoduladores (interferón beta y acetato de glatirámero) en un modelo murino de esclerosis múltiple / A comparative study of melatonin and immunomodulatory therapy with interferon beta and glatiramer acetate in a mouse model of multiple sclerosis

Introducción: La esclerosis múltiple (EM) es una enfermedad crónica desmielinizante autoinmune del sistema nervioso central (SNC) que produce neuroinflamación; un modelo es la encefalitis autoinmune experimental (EAE). La EM ha sido tratada con interferón beta (IFN-β) y acetato de glatirámero (AG). Se ha descrito que la melatonina (MLT) modula la respuesta del sistema inmune. El objetivo de este estudio fue observar el efecto de la administración de MLT contra los tratamientos de primera línea utilizados en la EM (IFN-β y AG).MétodosSe indujo EAE a ratas macho Sprague Dawley y se les administró IFN-β, AG o MLT. Se colectó líquido cefalorraquídeo (LCR) y se midieron citocinas proinflamatorias por multiplex, además del registro de la evaluación neurológica de la EAE.ResultadosTodos los animales inmunizados establecieron la EAE. Se evaluó el primer ciclo de recaída-remisión, observando que IFN-β y AG tienen mejores resultados que MLT en la evaluación clínica. La concentración en el LCR tanto de IL-1β como de IL-12p70 no se vio modificada por el modelo o por los tratamientos administrados. EL TNF-α se vio disminuido en el LCR por el IFN-β y la MLT bajo el modelo de EM.ConclusionesEs necesario realizar estudios posteriores para evaluar los mecanismos moleculares involucrados en el comportamiento de la MLT en la EAE, así como la cuantificación de otras citocinas en diferentes matrices biológicas para poder considerar la MLT como un agente antiinflamatorio regulador de la EM. (AU)

Introduction: Multiple sclerosis (MS) is a chronic, demyelinating, autoimmune disease of the central nervous system causing neuroinflammation. Experimental autoimmune encephalitis (EAE) is a model of the disease. MS is classically treated with interferon beta (IFN-β) and glatiramer acetate (GA). Melatonin (MLT) has been reported to modulate immune system responses. The aim of the present study is to analyse the effects of MLT administration in comparison with the first-line treatments for MS (IFN-β and GA).MethodsEAE was induced in male Sprague-Dawley rats; the animals subsequently received either IFN-β, GA, or MLT. Cerebrospinal fluid (CSF) samples were analysed by multiplex assay to determine the levels of proinflammatory cytokines. The neurological evaluation of EAE was also recorded.ResultsAll immunised animals developed EAE. We evaluated the first relapse-remission cycle, observing that IFN-β and GA had better results than MLT in the clinical evaluation. Neither EAE nor any of the treatments administered modified CSF IL-1β and IL-12p70 concentrations. However, IFN-β and MLT did decrease CSF TNF-α concentrations.ConclusionsFurther studies are needed to evaluate the molecular mechanisms involved in the behaviour of MLT in EAE, and to quantify other cytokines in different biological media in order for MLT to be considered an anti-inflammatory agent capable of regulating MS. (AU)

A comparative study of melatonin and immunomodulatory therapy with interferon beta and glatiramer acetate in a mouse model of multiple sclerosis. / Estudio comparativo de melatonina contra los tratamientos inmunomoduladores (interferón beta y acetato de glatirámero) en un modelo murino de esclerosis múltiple.

INTRODUCTION: Multiple sclerosis (MS) is a chronic, demyelinating, autoimmune disease of the central nervous system causing neuroinflammation. Experimental autoimmune encephalitis (EAE) is a model of the disease. MS is classically treated with interferon beta (IFN-ß) and glatiramer acetate (GA). Melatonin (MLT) has been reported to modulate immune system responses. The aim of the present study is to analyse the effects of MLT administration in comparison with the first-line treatments for MS (IFN-ß and GA). METHODS: EAE was induced in male Sprague-Dawley rats; the animals subsequently received either IFN-ß, GA, or MLT. Cerebrospinal fluid (CSF) samples were analysed by multiplex assay to determine the levels of proinflammatory cytokines. The neurological evaluation of EAE was also recorded. RESULTS: All immunised animals developed EAE. We evaluated the first relapse-remission cycle, observing that IFN-ß and GA had better results than MLT in the clinical evaluation. Neither EAE nor any of the treatments administered modified CSF IL-1ß and IL-12p70 concentrations. However, IFN-ß and MLT did decrease CSF TNF-α concentrations. CONCLUSIONS: Further studies are needed to evaluate the molecular mechanisms involved in the behaviour of MLT in EAE, and to quantify other cytokines in different biological media in order for MLT to be considered an anti-inflammatory agent capable of regulating MS.

Platelet-Rich Plasma for the Treatment of Photodamage of the Skin of the Hands. / Plasma rico en plaquetas en el tratamiento del fotodaño cutáneo en las manos.

BACKGROUND: Platelet-rich plasma (PRP) provides growth factors that stimulate fibroblast activation and induce the synthesis of collagen and other components of the extracellular matrix. The objective of this study was to evaluate the effect of PRP in the treatment of photodamage of the skin of the hands. MATERIAL AND METHODS: Experimental study enrolling persons with photoaged skin on the dorsum of the hands (Glogau photoaging scale, type III, or Fitzpatrick wrinkle classification, type II) were included between August 2012 and January 2013. A histological comparison was made of skin biopsies taken before and after the application of PRP to the skin of the dorsum of the hands. RESULTS: The mean (SD) age of the 18 women enrolled was 47.9 (4.3) years. Histological analysis showed an increase in the number of fibroblasts (P<.001), number of vessels (P<.001), and collagen density (P=.27). These changes produced significant improvements in the Fitzpatrick wrinkle and elastosis scale (P<.001) and in the Glogau photoaging scale (P=.01). CONCLUSIONS: PRP induced a reduction in the manifestations of skin aging, including an improvement in wrinkles and elastosis.

Evidence for widespread Leishmania infantum infection among wild carnivores in L. infantum periendemic northern Spain.

Leishmania spp. infection was investigated in tissue samples of wild carnivores from the Spanish Basque Country (BC), by PCR and DNA sequencing. The region is at the northern periphery of Leishmania infantum endemic Iberian Peninsula and infection in the dog (reservoir) or other species has not been previously reported. Leishmania kinetoplast DNA was detected by real-time PCR (rtPCR) in 28% (44/156) of animals. Specifically, in 26% of Eurasian badgers (n=53), 29% of foxes (n=48), 29% of stone martens (n=21) and in 25-50% of less numerous species including genets, wild cats, pole cats, European mink and weasels. Infected animals particularly badgers, were most prevalent in the southernmost province of the BC (Araba) in areas dominated by arable land. Subsequent amplification and sequencing of a fragment of the rRNA internal transcribed spacer 2 (ITS2) from a subset of rtPCR positives samples confirmed the species as L. infantum, showing a high sequence homogeneity with ITS2 sequences of L. infantum from dogs and humans from southern Spain. In summary, this study reports for the first time L. infantum infection in wild carnivores from the BC including in stone martens, pole cats and minks in which infection has not been previously described. It supports the need to study infection in dogs and people in this region and is an example of the value of infection surveillance in wildlife to assess potential risks in the domestic environment and their role in spreading infections in non-endemic areas.

Plasma fingerprinting with GC-MS in acute coronary syndrome.

New biomarkers of cardiovascular disease are needed to augment the information obtained from traditional indicators and to illuminate disease mechanisms. One of the approaches used in metabolomics/metabonomics for that purpose is metabolic fingerprinting aiming to profile large numbers of chemically diverse metabolites in an essentially nonselective way. In this study, gas chromatography-mass spectrometry was employed to evaluate the major metabolic changes in low molecular weight plasma metabolites of patients with acute coronary syndrome (n = 9) and with stable atherosclerosis (n = 10) vs healthy subjects without significant differences in age and sex (n = 10). Reproducible differences between cases and controls were obtained with pattern recognition techniques, and metabolites accounting for higher weight in the classification have been identified through their mass spectra. On this basis, it seems inherently plausible that even a simple metabolite profile might be able to offer improved clinical diagnosis and prognosis, but in addition, specific markers are being identified.

New perspective of diabetes response to an antioxidant treatment through metabolic fingerprinting of urine by capillary electrophoresis.

Benefits of capillary electrophoresis to provide a comprehensive snapshot of multiple metabolites in biological samples have been exploited. Afterwards, multivariate statistical methods can be employed in order to mine additional information from the data. Urine fingerprints of control and diabetic rats have shown the clear effects of an antioxidant treatment on diabetic animals, which were not seen in controls, in a rapid, simple and cost-effective way without identifying a single marker. The procedure involves the measurement of samples with a relatively inexpensive tool such as CE-UV, without any previous treatment other than filtration and the application of chemometric tools [PCA (principal components analysis) and PLS-DA (partial least squares discriminant analysis)]. Data pre-treatment of electrophoretic profiles (alignment, normalization and baseline correction) has shown to be key for further chemometric treatment. Once developed, the methodology can easily be applied for a rapid in vivo screening of extracts with potential in vitro activity. Classification was supported by that produced after PCA and PLS-DA of target variables obtained with selectively designed, time and reagent consuming methods.

Tocopherol fate in plasma and liver of streptozotocin-treated rats that orally received antioxidants and Spirulina extracts.

Streptozotocin-induced diabetic rats constitute a model of oxidative stress, and vitamin E continues to be a topic of speculation in this area. On the other hand, marine extracts, particularly microalgae extracts obtained with environmentally clean technologies and which demonstrate antioxidant activity in vitro, are a potential source of in vivo antioxidant defense. We have studied the alpha-tocopherol content in the plasma and liver of diabetic rats after 7 and 14 days under the condition, and before and after the treatment with vitamin E and C, as well as with different Spirulina extracts, as compared with the corresponding controls. The improvement of analytical methodology related to the determination of alpha-tocopherol in the plasma and liver of rats was also considered. To do this, a method previously developed for plasma, employing a single extraction step, was adapted and validated for liver after minor modifications. Moreover, stability of alpha-tocopherol in plasma of diabetic and control animals was compared in different storage conditions. Results showed that diabetic plasma strongly influences stability of alpha-tocopherol, even at -20 degrees C, but samples are stable for at least one year at -80 degrees C. Finally, regarding supplementation, results indicate that supplementation with alpha-tocopherol increases stored alpha-tocopherol in liver, but not in plasma, but this availability is strongly dependent on the stage of diabetes of the animal. Extracts of Spirulina platensis, despite showing antioxidant activity in vitro, increased alpha-tocopherol concentration in neither plasma nor liver.

Capillary electrophoresis of glutathione to monitor oxidative stress and response to antioxidant treatments in an animal model.

Glutathione plays a central role in metabolism and antioxidant defence. Several factors can influence the analytical efficiency and rapidity of the quantitative determination of glutathione. Procedures in sample pre-treatment have been compared in order to minimize analytical errors. Capillary electrophoresis has been chosen as a more adequate technique for obtaining a rapid and simple method for glutathione and glutathione disulfide determination in the blood and liver of the rat. The methods, once optimised, have been validated and applied for monitoring the oxidative stress in an animal model, such as the rat made diabetic by streptozotocin injection, when the animals are treated with antioxidants and compared with the corresponding controls.

Evaluation of diabetes-related short-chain organic acids in rat plasma by capillary electrophoresis.

A capillary zone electrophoresis method was optimised to analyse low-molecular-mass organic acids for the purpose of monitoring diabetes in rat plasma. The method included acetoacetic, 2-hydroxybutyric, lactic and uric acids. A variation in the background electrolyte allowed us to measure pyruvic acid in the same sample. Conditions have been optimised for measuring a large number of plasma samples corresponding to control and diabetic rats. Samples were mixed with acetonitrile (1:1, v/v) to precipitate proteins, centrifuged, diluted and injected. Tropic acid was chosen as an adequate internal standard. Separation was developed with reversed voltage by using a column cartridge pre-treated with polyacrylamide. Two electrophoretic buffers were employed: 0.150 M H3PO4 made up pH 6.20 with NaOH and 0.3 mM CaCl2 for acetoacetic, hydroxybutyric, lactic and uric acids, and 200 mM phosphate-10 mM acetate pH 4.0 for pyruvic acid, both with direct detection at 200 nm. The method was validated for linearity, accuracy and precision and the limits of quantification were calculated. The method was successfully applied to analyse these organic acids in control and diabetic animals. Acetoacetic and hydroxybutyric acids were clearly increased in diabetic rats, meanwhile no statistically significant difference has been found with the other acids.

Spontaneous micronuclei in peripheral blood erythrocytes from 35 mammalian species.

In this paper we report the results of a study to determine the frequencies of spontaneous micronucleated erythrocytes (MNE) in peripheral blood of 35 mammalian species. The main goal was to find mammals with a high spontaneous frequency of MNE that could, therefore, be good candidates for biomonitoring genotoxic agents in their natural habitat. We obtained 187 peripheral blood samples, but in 13 of the species we could only sample one individual. A wide range in the number of MNE (1/434 to 0/40,000 erythrocytes) was observed. Since the number of individuals per species is not high enough, this results should be cautiously considered. The cat, mouse, giraffe, pig, opossum and capuchin monkey seem to be suitable species for biomonitoring for genotoxic events.