ApTOLL: A new therapeutic aptamer for cytoprotection and (re)myelination after multiple sclerosis.

BACKGROUND AND PURPOSE: ApTOLL is an aptamer selected to antagonize toll-like receptor 4 (TLR4), a relevant actor for innate immunity involved in inflammatory responses in multiple sclerosis (MS) and other diseases. The currently available therapeutic arsenal to treat MS is composed of immunomodulators but, to date, there are no (re)myelinating drugs available in clinics. In our present study, we studied the effect of ApTOLL on different animal models of MS. EXPERIMENTAL APPROACH: The experimental autoimmune encephalomyelitis (EAE) model was used to evaluate the effect of ApTOLL on reducing the inflammatory component. A more direct effect on oligodendroglia was studied with the cuprizone model and purified primary cultures of murine and human oligodendrocyte precursor cells (OPCs) isolated through magnetic-activated cell sorting (MACS) from samples of brain cortex. Also, we tested these effects in an ex vivo model of organotypic cultures demyelinated with lysolecithin (LPC). KEY RESULTS: ApTOLL treatment positively impacted the clinical symptomatology of mice in the EAE and cuprizone models, which was associated with better preservation plus restoration of myelin and oligodendrocytes in the demyelinated lesions of animals. Restoration was corroborated on purified cultures of rodent and human OPCs. CONCLUSION AND IMPLICATIONS: Our findings reveal a new therapeutic approach for the treatment of inflammatory and demyelinating diseases such as MS. The molecular nature of the aptamer exerts not only an anti-inflammatory effect but also neuroprotective and remyelinating effects. The excellent safety profile demonstrated by ApTOLL in animals and humans opens the door to future clinical trials in MS patients.

Adult intestinal malrotation: imaging findings.

Intestinal malrotation is the most common congenital anomaly of the small intestine. However, it is associated with delayed diagnosis due to the lack of specificity of its symptoms, which can lead to devastating consequences such as intestinal volvulus or massive intestinal necrosis. We present a clinical case in which we highlight the importance of abdominal computed tomography and the detection of its characteristic signs for the early identification of this pathology.

Revealing the 1H-NMR Profiling of Six Edible Mushrooms Consumed in the Northeastern Highlands of Puebla, Mexico.

The 1H-NMR metabolomics profiling of six edible mushrooms consumed in the northeastern highlands of Puebla, Mexico is presented. These fungi were morpho- and molecularly identified as Infundibulicybe squamulosa, Amanita jacksonii, Lepista nuda, Russula delica, Russula brevipes, and Lactarius indigo. The chemical profiling confirmed the presence of eight essential amino acids and their derivatives, six organic acids, six nucleosides, low amounts of reducing sugars, and valuable nutraceuticals such as betaine, carnitine, glycero-3-phosphocholine and O-acetylcarnitine which were differentially determined and quantified in the six mushrooms by qNMR. Principal component analysis (PCA) and orthogonal projections to latent structures discriminant analysis (OPLS-DA) generated four different groups. Two of these groups were constituted by fungal species with phylogenic relationships whereas non-phylogenetic related species were separated from each other. The potential use of 1H-NMR metabolomics and chemometrics to group macromycetes and determine the nutritional and nutraceutical potential of these local foods is demonstrated.

Diazoxide and moderate-intensity exercise improve skeletal muscle function by decreasing oxidants and enhancing antioxidant defenses in hypertensive male rats.

High sodium intake is decisive in the incidence increase and prevalence of hypertension, which has an impact on skeletal muscle functionality. Diazoxide is an antihypertensive agent that inhibits insulin secretion and is an opener of KATP channels (adosine triphosphate sensitive potasium channels). For this reason, it is hypothesized that moderate-intensity exercise and diazoxide improve skeletal muscle function by reducing the oxidants in hypertensive rats. Male Wistar rats were assigned into eight groups: control (CTRL), diazoxide (DZX), exercise (EX), exercise + diazoxide (EX + DZX), hypertension (HTN), hypertension + diazoxide (HTN + DZX), hypertension + exercise (HTN + EX), and hypertension + exercise + diazoxide (HTN + EX + DZX). To induce hypertension, the rats received 8% NaCl dissolved in water orally for 30 days; in the following 8 weeks, 4% NaCl was supplied to maintain the pathology. The treatment with physical exercise of moderate intensity lasted 8 weeks. The administration dose of diazoxide was 35 mg/kg intraperitoneally for 14 days. Tension recording was performed on the extensor digitorum longus and the soleus muscle. Muscle homogenates were used to measure oxidants using fluorescent probe and the activity of antioxidant systems. Diazoxide and moderate-intensity exercise reduced oxidants and increased antioxidant defenses.

Efficacy of a benzothiazole-based LRRK2 inhibitor in oligodendrocyte precursor cells and in a murine model of multiple sclerosis.

AIMS: Multiple sclerosis (MS) is a chronic neurological disease that currently lacks effective curative treatments. There is a need to find effective therapies, especially to reverse the progressive demyelination and neuronal damage. Oligodendrocytes form the myelin sheath around axons in the central nervous system (CNS) and oligodendrocyte precursor cells (OPCs) undergo mechanisms that enable spontaneously the partial repair of damaged lesions. The aim of this study was to discover small molecules with potential effects in demyelinating diseases, including (re)myelinating properties. METHODS: Recently, it has been shown how LRRK2 inhibition promotes oligodendrogliogenesis and therefore an efficient repair or myelin damaged lesions. Here we explored small molecules inhibiting LRRK2 as potential enhancers of primary OPCs proliferation and differentiation, and their potential impact on the clinical score of experimental autoimmune encephalomyelitys (EAE) mice, a validated model of the most frequent clinical form of MS, relapsing-remitting MS. RESULTS: One of the LRRK2 inhibitors presented in this study promoted the proliferation and differentiation of OPC primary cultures. When tested in the EAE murine model of MS, it exerted a statistically significant reduction of the clinical burden of the animals, and histological evidence revealed how the treated animals presented a reduced lesion area in the spinal cord. CONCLUSIONS: For the first time, a small molecule with LRRK2 inhibition properties presented (re)myelinating properties in primary OPCs cultures and potentially in the in vivo murine model. This study provides an in vivo proof of concept for a LRRK2 inhibitor, confirming its potential for the treatment of MS.

Podiatric Skin and Nail Involvement and Biomechanical Pathology in Renal Transplant Recipients: Assessment of the Foot as a Contributing Factor to Their Health.

Although several studies show the prevalence of podiatric conditions in people with end-stage renal disease or renal replacement therapy with hemodialysis, there is little scientific literature on this when subjects are undergoing kidney transplantation. The aim of this study is to determine the prevalence of podiatric skin and nail pathology in renal transplant recipients. A descriptive, observational, prevalence study was conducted at the Nephrology Department of the University Hospital of A Coruña. A total of 371 subjects were studied. The variables studied were sociodemographic (age, sex), anthropometric (Body Mass Index), comorbidity (Charlson Comorbidity Index), and podological (skin and nail alterations). A high presence of skin (83.1%) and nail pathology (85.4%) was observed, with hyperkeratosis (68.8%), onychogryphosis (39.4%), and onychocryptosis (36.9%) being the most predominant alterations. Although it was not significant, patients with a higher risk of presenting podiatric pathology were of female sex and had a high BMI, and both age and the Charlson comorbidity index were significantly associated with this risk. There was an increased risk of both skin and nail pathology at older age and in the presence of diabetes mellitus.

Comparison of the Aptima MG and Cobas TV/MG tests for the detection of Mycoplasma genitalium in urogenital and extragenital samples.

BACKGROUND: Mycoplasma genitalium (M. genitalium) is an emerging sexually transmitted pathogen of increasing importance. The objective of this study was to compare two tests for the detection of M. genitalium; the Aptima® MG test (Hologic® Inc., San Diego, CA) and the Cobas® TV/MG test (Roche® Diagnostics, Mannheim, Germany). METHODS: This is a prospective descriptive study where a total of 489 genital and extragenital samples were analyzed in parallel and in random order by both systems. The samples were collected from patients attending the Sexually Transmitted Infections Center in Seville and the Infectious Diseases consultation of the Virgen de Valme Hospital. RESULTS: The overall agreement between both trials was very good (k > 0.91). The sensitivity and specificity of the Aptima® MG test were 100% and 98.7% respectively for the Cobas® TV/MG test. CONCLUSION: Both systems showed excellent performance for the detection of M. genitalium.

Atypical ocular Chlamydia trachomatis infections in two patients attending in a second-level hospital: a case report.

AIM: To report two atypical inclusion conjunctivitis cases due to Chlamydia trachomatis in young adults. METHOD: Transcription mediated amplification for C. trachomatis was performed using Aptima Combo 2 Assay (Hologic, Spain). RESULTS: The first patient was managed as an orbital disorder because he had unilateral location, and ptosis was observed. Orbital nuclear magnetic resonance revealed normal results, and conjunctival biopsy did not indicate significant results. For the second patient, thyroid eye disease was suspected, but the orbital nuclear magnetic resonance revealed normal results. Conjunctival exudate samples were collected and sent to the Microbiology Laboratory where C. trachomatis was confirmed. Both patients demonstrated a great improvement with oral azithromycin 1 g. CONCLUSION: Inclusion conjunctivitis could present as unspecified unilateral or bilateral chronic conjunctivitis. Thus, suspecting it would be important in order to prevent spread and wasting diagnostic resources.

Microorganisms and Antibiotic Resistance of Bacterial Keratitis at a Rural County Hospital in Seville.

PURPOSE: To review the incidence, resistance patterns, and management of bacterial keratitis during the past 4 years. METHODS: We retrospectively reviewed the clinical records of microbiological isolates from patients with a clinical diagnosis of bacterial keratitis. RESULTS: A total of 159 patients were analyzed, and 102 microorganisms were isolated from 129 cultures. In these cultures, 23.7% of the microorganisms were gram positive, 60.8% were gram negative, and 15.5% were fungi. Pseudomonas aeruginosa was the most common bacteria (9.2%), followed by Serratia marcescens (4.4%) and Staphylococcus aureus (4%). Resistance to fluoroquinolones and aminoglycosides was found to be 23.1% and 53.1% in gram-positive and 2.8% and 13.9% in gram-negative bacteria, respectively. Resistance to ceftazidime against gram-negative bacteria was 13.9%. No resistance to vancomycin was observed. CONCLUSIONS: A high resistance rate to aminoglycosides and fluoroquinolones was observed in gram-positive bacteria. We concluded that fluoroquinolones or aminoglycosides may not be suitable for initial monotherapy in patients with severe bacterial keratitis.

Enfermedad arterial periférica en pacientes trasplantados renales. Validez del cuestionario de Edimburgo para el diagnóstico de enfermedad / Peripheral arterial disease in renal transplant patients. Validity of the Edinburgh questionnaire for disease diagnosis

RESUMEN Objetivos: Determinar en pacientes trasplantados renales la prevalencia de enfermedad arterial periférica y la validez de las manifestaciones clínicas de claudicación intermitente para su diagnóstico. Material y métodos: Ámbito y período: Servicio de Nefrología del Complejo Hospitalario Universitario A Coruña, 2013-2017. Criterios inclusión: pacientes trasplantados renales con injerto funcionante que consientan participar en el estudio. Justificación del tamaño muestral: n=371 pacientes (seguridad= 95% y precisión= ± 4,25%). Mediciones: edad, edad al trasplante, sexo, dislipemia, índice de masa corporal, tabaquismo, diabetes, signo de Godet, edema perimaleolar, índice tobillo-brazo y cuestionario Edimburgo. El riesgo cardiovascular se midió mediante los scores Framingham-Wilson, Regicor, SCORE y Dorica. Resultados: La edad media al trasplante fue de 47,86 ± 12,62 años, 65,5% hombres. El 8,7% de los pacientes presentan un índice tobillo-brazo <0,90. El 16,2% de los pacientes manifiestan claudicación intermitente según el cuestionario Edimburgo. La concordancia entre ambas pruebas diagnósticas es débil (índice de Kappa=0,34). El cuestionario de Edimburgo mostró sensibilidad del 59,38% para predecir índice tobillo-brazo <0,90 y especificidad del 88,10%. Las variables asociadas a la presencia de arteriopatía son la edad al trasplante (OR=1,07) y el tabaquismo (OR=6,17), encontrándose la dislipemia en el límite de la significación estadística. Conclusiones: Una décima parte de los pacientes presentan arteriopatía periférica. La concordancia entre el cuestionario Edimburgo y el índice tobillo-brazo es débil. Por lo que debería usarse el índice tobillo-brazo como método diagnóstico. Las manifestaciones clínicas infraestiman la prevalencia de arteriopatía. La edad, el tabaquismo y la dislipemia incrementan su riesgo. Los pacientes con arteriopatía presentan riesgo cardiovascular más elevado.

ABSTRACT Objectives: To assess the prevalence of peripheral artery disease in kidney transplant patients and the validity of intermittent claudication for its diagnosis. Methods: Setting and period: Nephrology Department of the University Hospital A. Coruña, 2013-2017. Inclusion criteria: transplant patients with functioning grafts who gave their consent to participate in the study. Sample size rationale: n=371 patients (confidence interval= 95%; precision= ± 4.25%). Measurements: age, age at the time of transplant, sex, dyslipemia, body mass index, smoking, diabetes, sign of Godet, perimalleolar edema, ankle-brachial index and the Edinburgh Questionnaire. Cardiovascular risk was measured with these scores: Framingham-Wilson, Regicore, SCORE and Dorica. Results: The mean age at the time of transplant was 47.86±12.62; 65.5% of patients were men and 8.7% of them had an ankle-brachial index of <0.90. When answering the Edinburgh Questionnaire, 16.2% of subjects reported suffering from intermittent claudication. Concordance between these two diagnostic tests is poor (kappa index= 0.34). The Edinburgh Questionnaire showed a sensitivity of 59.38% in predicting the ankle-brachial index (<0.90) and specificity (88.10%). The variables associated with the presence of artery disease are age at the time of transplant (OR=1.07) and smoking (OR=6.17), dyslipidemia being at the limit of statistical significance. Conclusions: A tenth part of the patients have peripheral artery disease. Concordance between the Edinburgh Questionnaire and the ankle-brachial index is poor; therefore, the latter should be used as diagnostic method. Clinical signs and symptoms underestimate the prevalence of artery disease. Age, smoking and dyslipidemia increase the risk of this disease. Artery disease patients have a higher cardiovascular risk.

Discordance of the Evaluation of the Plantar Footprint in Kidney Transplant.

BACKGROUND: The measures used for the diagnosis of biomechanical pathologic disorders are variable. The aim of this study was to determine the concordance between the Clarke angle and the Chippaux-Smirak and Staheli indices in evaluation of the footprints of kidney transplant patients. METHODS: An observational prevalence study was performed in kidney transplant patients in the University Hospital Complex A Coruña from 1981 through 2014. After obtaining patient informed consent and ethics committee approval, sociodemographic (age and sex) and anthropometric (body mass index) variables and comorbidity (Charlson Comorbidity Index score) were assessed for each patient, and a podiatric medical examination was performed of the footprint through a pedigree (Chippaux-Smirak index, Staheli index, and Clarke angle). A descriptive analysis of all of the variables was performed; for agreement, the kappa index and the Bland-Altman method were used. RESULTS: We found a predominance of male sex (65.6%) and a high level of overweight (43.3%) in the sample. The highest concordance between the indices studied is presented by the Chippaux-Smirak index, with a kappa index of 0.86 for the left foot and 0.76 for the right foot. CONCLUSIONS: High agreement was attained among observers in measurements of the footprints of kidney transplant patients.

Effects of Apocynin on Heart Muscle Oxidative Stress of Rats with Experimental Diabetes: Implications for Mitochondria.

Diabetes mellitus (DM) constitutes one of the public health problems today. It is characterized by hyperglycemia through a defect in the ß-cells function and/or decreased insulin sensitivity. Apocynin has been tasted acting directly as an NADPH oxidase inhibitor and reactive oxygen species (ROS) scavenger, exhibiting beneficial effects against diabetic complications. Hence, the present study's goal was to dissect the possible mechanisms by which apocynin could mediate its cardioprotective effect against DM-induced oxidative stress. Male Wistar rats were assigned into 4 groups: Control (C), control + apocynin (C+A), diabetes (D), diabetes + apocynin (D+A). DM was induced with streptozotocin. Apocynin treatment (3 mg/kg/day) was applied for 5 weeks. Treatment significantly decreased blood glucose levels and insulin resistance in diabetic rats. In cardiac tissue, ROS levels were higher, and catalase enzyme activity was reduced in the D group compared to the C group; the apocynin treatment significantly attenuated these responses. In heart mitochondria, Complexes I and II of the electron transport chain (ETC) were significantly enhanced in the D+A group. Total glutathione, the level of reduced glutathione (GSH) and the GSH/ oxidized glutathione (GSSG) ratio were increased in the D+A group. Superoxide dismutase (SOD) and the glutathione peroxidase (GSH-Px) activities were without change. Apocynin enhances glucose uptake and insulin sensitivity, preserving the antioxidant defense and mitochondrial function.

Prolactin modifies the in vitro LPS-induced chemotactic capabilities in human fetal membranes at the term of gestation.

PROBLEM: Immune responses of fetal membranes involve the production of chemoattractant mediators causing infiltration of maternal and fetal leukocytes, intrauterine inflammation and potentially the disruption of maternal-fetal tolerance. Prolactin (PRL) has deep immunoregulatory effects in the fetal-maternal interface. We aimed to test the in vitro PRL effect upon chemotactic capacities of human fetal membranes. METHOD OF STUDY: Fetal membranes and umbilical cord blood were collected from healthy non-laboring caesarean deliveries at term. Fetal membranes were cultured in Transwell® frames to mimic the barrier function between choriodecidual and amniotic sides. Tissues were treated with PRL, Lipopolysaccharide (LPS), or both simultaneously. Then, RANTES, MCP-1, MIP-1α, IP-10, and PECAM-1 were quantified in a conditioned medium by choriodecidual or amniotic sides. The chemotaxis of subsets of migrating mononuclear cells from umbilical cord blood was evaluated in a Boyden Chamber in response to the conditioned medium by both sides. RESULTS: Lipopolysaccharide stimulates the production of RANTES, MCP-1, MIP-1α, and PECAM-1 in choriodecidua, while MIP-1α and PECAM-1 only increase in amnion. PRL decrease RANTES, MCP-1, and MIP-1 only in choriodecidua, but PECAM-1 was decreased mainly in amnion. The leukocyte migration was regulated significantly in response to the conditioned medium by the amnion, increase in the conditioned medium after LPS treatment, contrary with, the leukocyte migration decreased in a significant manner in response to conditioned medium after PRL and LPS-PRL co-treatment. Finally, T cells were the most responsive subset of cells. CONCLUSIONS: Prolactin modified in a tissue-specific manner the chemotactic factor and the leukocyte migration differentially in fetal membranes.

Serum angiopoietin-like 3 levels are elevated in obese non diabetic men but are unaffected during an oral glucose tolerance test.

This study aimed to determine ANGPTL3 serum levels in healthy young lean and obese non-diabetic men during an oral glucose tolerance test (OGTT) and correlate them with anthropometric, biochemical and hormonal parameters. A case-control study was carried out and 30 young obese non-diabetic (23.90 ± 3.84 years and BMI 37.92 ± 4.85 kg/m2) and 28 age-matched healthy lean (24.56 ± 3.50 years and BMI of 22.10 ± 1.72 kg/m2) men were included in this study. The primary outcome measures were serum basal ANGPTL3 and ANGPTL3-area under the curve (AUC) levels. The percentage of body fat was measured by dual-energy X-ray absorptiometry and biochemical, hormonal and insulin resistance indices were determined. Basal ANGPTL3 and ANGPTL3-AUC levels were significantly elevated (p < 0.05) in young obese subjects compared with lean subjects and were positively and significantly associated with different anthropometric measurements. Fasting ANGPTL3 serum levels were positively correlated with fasting insulin, leptin, Leptin/Adiponectin index and triglyceride-glucose index. Moreover, ANGPTL3-AUC was negatively correlated with Matsuda index. In this regard, chronically high ANGPTL3 levels in young obese subjects might favor triglyceride-rich lipoprotein clearance to replenish triglyceride stores by white adipose tissue rather than oxidative tissues.

Evaluation of 2 Commercial Assays for the Detection of Lymphogranuloma Venereum in Rectal Samples.

BACKGROUND: The early identification of the Chlamydia trachomatis variants that cause lymphogranuloma venereum (LGV) is very important to establish an adequate antibiotic treatment. This identification should be made with molecular techniques that are easy to perform and accessible to most microbiology laboratories. The objective of this study was to evaluate 2 real-time polymerase chain reaction (PCR)-based assay (VIASURE Haemophilus ducreyi + C. trachomatis (LGV) real-time PCR detection kit and the Allplex Genital ulcer Assay) for the detection of LGV in rectal samples. MATERIALS AND METHODS: Prospective study on positive rectal samples for C. trachomatis. All samples were processed in parallel by both tests. As a molecular reference method and to solve possible discrepancies between both assays, a PCR-based restriction fragment length polymorphism analysis of the major outer membrane protein gene (omp1) was used. RESULTS: In total, we detected 157 positive rectal samples for C. trachomatis, of which 36 were identified as LGV by PCR-based restriction fragment length polymorphism analysis. The positive percent agreement, negative percent agreement, and overall percent agreement were 88.9%, 100%, and 97.3%, respectively, for the Allplex Genital ulcer assay and 91.6%, 100%, and 97.1%, respectively, for the VIASURE assay. In the direct comparison between the Seegene assay and the VIASURE assay, we obtained a kappa concordance index of 0.98 between both tests. CONCLUSIONS: According to the results obtained, both tests could be used for the detection of LGV in rectal samples.

Updating of in vitro antifungal susceptibility tests. / Actualización de los métodos de estudio de sensibilidad in vitro a los antifúngicos.

Fungal diseases, including those caused by (multi)drug-resistant fungi, still represent a global public health concern. Information on the susceptibility of these microorganisms to antifungal agents must be quickly produced to help clinicians initiate appropriate antifungal therapies. Unfortunately, antifungal susceptibility tests are not as developed or widely implemented as antibacterial tests, being similar in design, accuracy and reproducibility, but also laborious and slow. In this article, we review the methods of in vitro susceptibility testing, both reference (CLSI and EUCAST), commercial and new methods based on proteomics (MALDI-TOF MS) and in the detection of resistance genes by nucleic acid amplification techniques. In addi-tion, we discuss the newly established clinical breakpoints, as well as the epidemiological cut-off points, which constitute a new category that can help in the early identification of isolates that have acquired resistance mechanisms. We also discuss the advantages and limitations of each of the methods studied. Therefore, we can conclude that, although there has been much progress in studies of in vitro susceptibility testing to antifungals, there are still limitations in its application in the daily routine of microbiology labo-ratories, although it seems that the future is promising with the new technologies based on proteomics and nucleic acid amplification. Supplement information: This article is part of a supplement entitled «SEIMC External Quality Control Programme. Year 2016¼, which is sponsored by Roche, Vircell Microbiologists, Abbott Molecular and Francisco Soria Melguizo, S.A. © 2019 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosasy Microbiología Clínica. All rights reserved.

Verbal intelligence in bilinguals when measured in L1 and L2.

This study was aimed at studying the Verbal IQ in two groups of Spanish/English bilinguals: simultaneous and early sequential bilinguals. 48 Spanish/English bilinguals born in the U.S. or Latin American countries but moving to United States before the age of 10 were selected. The verbal subtests of the Wechsler Adult Intelligence Scale (English and Spanish) - Third Edition (WAIS-III) was administered. Overall, performance was significantly better in English for both groups of bilinguals. Verbal IQ difference when tested in Spanish and English was about one standard deviation higher in English for simultaneous bilinguals, and about half standard deviation for early sequential bilinguals. In both groups, Verbal IQ in English was about 100; considering the level of education of our sample (bachelor degree, on average), it can be assumed that Verbal IQ in English was lower than expected, suggesting that bilinguals may be penalized even when evaluated in the dominant language.