A dual PCR-based sequencing approach for the identification and discrimination of Echinococcus and Taenia taxa.
Boubaker G, Marinova I, Gori F, Hizem A, Müller N, Casulli A, et al. Mol Cell Probes. 2016 May 27. pii: S0890-8508(16)30038-X.
DOI: 10.1016/j.mcp.2016.05.004. [Epub ahead of print]

Reliable and rapid molecular tools for the genetic identification and differentiation of Echinococcus species and/or genotypes are crucial for studying spatial and temporal transmission dynamics. Here, we describe a novel dual PCR targeting regions in the small (rrnS) and large (rrnL) subunits of mitochondrial ribosomal RNA (rRNA) genes, which enables (i) the specific identification of species and genotypes of Echinococcus (rrnS + L-PCR) and/or (ii) the identification of a range of taeniid cestodes, including different species of Echinococcus, Taenia and some others (17 species of diphyllidean helminths). This dual PCR approach was highly sensitive, with an analytical detection limit of 1 pg for genomic DNA of Echinococcus. Using concatenated sequence data derived from the two gene markers (1225 bp), we identified five unique and geographically informative single nucleotide polymorphisms (SNPs) that allowed genotypes (G1 and G3) of Echinococcus granulosus sensu stricto to be distinguished, and 25 SNPs that allowed differentiation within Echinococcus canadensis (G6/7/8/10). In conclusion, we propose that this dual PCR-based sequencing approach can be used for molecular epidemiological studies of Echinococcus and other taeniid cestodes.

Amygdala electrical stimulation inducing spatial memory recovery produces an increase of hippocampal bdnf and arc gene expression.
Mercerón-Martínez D, Almaguer-Melian W, Alberti-Amador E, Estupiñán B, Fernández I, Bergado JA. Brain Res Bull.
2016 Jun 1;124:254–61. DOI: 10.1016/j.brainresbull.2016.05.017. [Epub ahead of print]

Amygdala seems to promote the consolidation of plastic modification in different brain areas and these long-term brain changes require a rapid de novo RNA and protein synthesis. We have previously shown that basolateral amygdala electrical stimulation produces a partial recovery of spatial memory in fimbria-fornix lesioned animals and it is also able to increase the BDNF protein content in the hippocampus. The emerging question is whether these increased BDNF protein content arises from previously synthesized RNA or from de novo RNA expression. Now we address the question if amygdala electrical stimulation 15min after daily water maze training produces a rapid de novo RNA synthesis in the hippocampus, a critical brain area for spatial memory recovery in fimbria-fornix lesioned animals. In addition, we also study RNA arc expression, a gene which is essential for memory and neural plasticity processes. To this purpose, we study amygdala stimulation effects on the expression of plasticity related-early-genes bdnf and arc in the hippocampus of fimbria-fornix lesioned animals trained in a water-maze for 4 days. We also checked on the expression of both genes in non-lesioned, untrained animals (acute condition) at 0.5, 1, 2 and 24h after basolateral amygdala electrical stimulation. Our data from trained animals confirm that daily amygdala electrical stimulation 15min after water maze training produces a partial memory recovery and that is coupled to an increase of bdnf and arc genes expression in the hippocampus. Additionally, the acute study shows that a single session of amygdala stimulation induces a transient increase of both genes (peaking at 30min). These results confirm the memory improving effect of amygdala stimulation in fimbria-fornix-lesioned animals and sustain the assumption that the memory improving effect is mediated by newly synthetized BDNF acting on a memory relevant structure like the hippocampus. The increased amount of BDNF within the hippocampus seems to be locally synthetized by mechanisms activated by the amygdala stimulation.

APL-1, an altered peptide ligand derived from heat-shock protein, alone or combined with methotrexate attenuates murine collagen-induced arthritis.
Lorenzo N, Altruda F, Silengo L, Del Carmen Dominguez M. Clin Exp Med. 2016 May 9. [Epub ahead of print]

Induction of tolerance to autoantigens in vivo is a complex process that involves several mechanisms such as the induction of regulatory T cells and changes in the cytokine and chemokine profiles. This approach represents an attractive alternative for treatment of autoimmune diseases. APL-1 is an altered peptide ligand derived from a novel CD4 + T cell epitope of human heat-shock protein of 60 kDa (HSP60), an autoantigen involved in the pathogenesis of rheumatoid arthritis (RA). We have shown previously that this peptide efficiently inhibited the course of adjuvant-induced arthritis in Lewis rats and induced regulatory T cell (Treg) in ex vivo assay with PBMC isolated from RA patients. This study was undertaken to evaluate the therapeutic effect of APL-1 and its combination with methotrexate (MTX) in collagen-induced arthritis (CIA). CIA was induced in male DBA/1 mice at 8 weeks of age by immunization with chicken collagen. APL, MTX or both were administrated beginning from arthritis onset. Therapeutic effect was evaluated by arthritis and joint pathologic scores. In addition, TNFα and IL-10 in sera were measured by ELISA. Treg induction was assessed by FACS analysis. APL-1 inhibits efficiently the course of arthritis in CIA, similar to MTX. In addition, therapy with APL-1 plus MTX reduced CIA in mice, associated with an increase in Treg. These facts reinforce the therapeutic possibilities of APL-1 as a candidate drug for treatment of RA.

Avoiding a second amniocentesis to corroborate prenatal diagnosis by using refrigerated samples.
Méndez-Rosado LA, Cuétara E, Molina-Gamboa O, Suarez-Mayedo U, Huertas-Perez G, Barrios-Martinez A, et al. J Maternfetal Neonatal Med. 2016 May 26:1–5. [Epub ahead of print].

Objective This study is aimed to probe the usefulness of refrigerated aliquots of amniotic fluid to be used for fluorescence in situ hybridization (FISH) in order to perform an accurate prenatal diagnosis avoiding the risk related to an additional amniocentesis procedure and the psychological stress to the pregnant woman and her family. Methods Non-cultured amniotic fluid (AF) samples were analyzed by FISH. Such samples were divided into two groups. The first one included fresh collected AF (FCAF, N=30). The second one included refrigerated samples of AF (RSAF, N=12) to corroborate uncertain chromosome aberration identification obtained by conventional methods. Sample refrigeration did not exceed 18 days. Results No differences were found between groups. In the RSAF group, three cases of chromosomal mosaicismo and seven cases of pseudomosaicism were corroborated. No alteration adjudicated to aberrant chromosomal line presence was found in born children according to genetic specialists’ criteria. In the two remaining cases, applied procedure allowed elucidating fetal sex in one case and the origin of a marker chromosome in the other. Conclusions Amniocytes obtained from RAFS are a useful biological material to be assayed by FISH, achieving an accurate prenatal diagnosis and avoiding an additional amniocentesis.

Current Developments on Optical Feedback Interferometry as an All-Optical Sensor for Biomedical Applications.
Perchoux J, Quotb A, Atashkhooei R, Azcona FJ, Ramírez-Miquet EE, Bernal O, et al. Sensors (Basel). 2016 May 13;16(5). pii: E694. DOI: 10.3390/s16050694.

Optical feedback interferometry (OFI) sensors are experiencing a consistent increase in their applications to biosensing due to their contactless nature, low cost and compactness, features that fit very well with current biophotonics research and market trends. The present paper is a review of the work in progress at UPC-CD6 and LAAS-CNRS related to the application of OFI to different aspects of biosensing, both in vivo and ex vivo. This work is intended to present the variety of opportunities and potential applications related to OFI that are available in the field. The activities presented are divided into two main sensing strategies: The measurement of optical path changes and the monitoring of flows, which correspond to sensing strategies linked to the reconstruction of changes of amplitude from the interferometric signal, and to classical Doppler frequency measurements, respectively. For optical path change measurements, measurements of transient pulses, usual in biosensing, together with the measurement of large displacements applied to designing palliative care instrumentation for Parkinson disease are discussed. Regarding the Doppler-based approach, progress in flow-related signal processing and applications in real-time monitoring of non-steady flows, human blood flow monitoring and OFI pressure myograph sensing will be presented. In all cases, experimental setups are discussed and results presented, showing the versatility of the technique. The described applications show the wide capabilities in biosensing of the OFI sensor, showing it as an enabler of low-cost, all-optical, high accuracy biomedical applications.

Early discoid lupus erythematosus protects against renal disease in patients with systemic lupus erythematosus: longitudinal data from a large Latin American cohort.
Pons-Estel GJ, Aspey LD, Bao G, Pons-Estel BA, Wojdyla D, Saurit V, et al. Lupus. 2016 May 26. pii: 0961203316651740. [Epub ahead of print]

Objectives The objective of this study was to examine whether early discoid lupus erythematosus (DLE) would be a protective factor for further lupus nephritis in patients with systemic lupus erythematosus (SLE). Methods We studied SLE patients from GLADEL, an inception longitudinal cohort from nine Latin American countries. The main predictor was DLE onset, which was defined as physician-documented DLE at SLE diagnosis. The outcome was time from the diagnosis of SLE to new lupus nephritis. Univariate and multivariate survival analyses were conducted to examine the association of DLE onset with time to lupus nephritis. Results Among 845 GLADEL patients, 204 (24.1%) developed lupus nephritis after SLE diagnosis. Of them, 10 (4.9%) had DLE onset, compared to 83 (12.9%) in the group of 641 patients that remained free of lupus nephritis (hazard ratio 0.39; P=0.0033). The cumulative proportion of lupus nephritis at 1 and 5 years since SLE diagnosis was 6% and 14%, respectively, in the DLE onset group, compared to 14% and 29% in those without DLE (P=0.0023). DLE onset was independently associated with a lower risk of lupus nephritis, after controlling for sociodemographic factors and disease severity at diagnosis (hazard ratio 0.38; 95% confidence interval 0.20–0.71). Conclusions Our data indicate that DLE onset reduces the risk of further lupus nephritis in patients with SLE, independently of other factors such as age, ethnicity, disease activity, and organ damage. These findings have relevant prognosis implications for SLE patients and their clinicians. Further studies are warranted to unravel the biological and environmental pathways associated with the protective role of DLE against renal disease in patients with SLE.

Efficient and biologically relevant consensus strategy for Parkinson’s disease gene prioritization.
Cruz-Monteagudo M, Borges F, Paz-Y-Miño C, Cordeiro MN, Rebelo I, Pérez-Castillo Y, et al. BMC Med Genomics. 2016 Mar 9;9(1):12 DOI: 10.1186/s12920-016-0173-x

Background The systemic information enclosed in microarray data encodes relevant clues to overcome the poorly understood combination of genetic and environmental factors in Parkinson’s disease (PD), which represents the major obstacle to understand its pathogenesis and to develop disease-modifying therapeutics. While several gene prioritization approaches have been proposed, none dominate over the rest. Instead, hybrid approaches seem to outperform individual approaches. Methods A consensus strategy is proposed for PD related gene prioritization from mRNA microarray data based on the combination of three independent prioritization approaches: Limma, machine learning, and weighted gene co-expression networks. Results The consensus strategy outperformed the individual approaches in terms of statistical significance, overall enrichment and early recognition ability. In addition to a significant biological relevance, the set of 50 genes prioritized exhibited an excellent early recognition ability (6 of the top 10 genes are directly associated with PD). 40 % of the prioritized genes were previously associated with PD including well-known PD related genes such as SLC18A2, TH or DRD2. Eight genes (CCNH, DLK1, PCDH8, SLIT1, DLD, PBX1, INSM1, and BMI1) were found to be significantly associated to biological process affected in PD, representing potentially novel PD biomarkers or therapeutic targets. Additionally, several metrics of standard use in chemoinformatics are proposed to evaluate the early recognition ability of gene prioritization tools. Conclusions The proposed consensus strategy represents an efficient and biologically relevant approach for gene prioritization tasks providing a valuable decision-making tool for the study of PD pathogenesis and the development of disease-modifying PD therapeutics.

Estimating and mapping the incidence of giardiasis in Colombia, 2009-2013.
Rodríguez-Morales AJ, Granados-Álvarez S, Escudero-Quintero H, Vera-Polania F, Mondragon-Cardona A, Díaz-Quijano FA, et al. Int J Infect Dis. 2016 Jun 13. pii: S1201-9712(16)31089-X.
DOI: 10.1016/j.ijid.2016.06.005. [Epub ahead of print]

Giardiasis is one of the most common intestinal infections in the world. In Colombia there are no national studies about its morbidity. In this study incidence rates of giardiasis between 2009 and 2013 were estimated. An observational, retrospective study of the giardiasis incidence in Colombia, 2009-2013, with data extracted from the so-called personal health records system (Registro Individual de Prestación de Servicios, RIPS) was performed. Official population estimates from the National Department of Statistics (DANE) were used for crude and adjusted incidence rates estimation (cases/100,000 pop). During the period studied, 15,851 cases were reported (median 3,233/year; 5-year cumulated crude national rate of 33.97 cases/100,000 pop); 50.3% were female; 58.4% were <10 years old and 14.8% were 10-19 years old. From the total, 17.7% were from Bogota (10.07 cases/100,000 pop, 2009), followed by Antioquia with 10.9% (9.42, 2009), Atlántico with 8.6% (15.67, 2009) and Risaralda with 6.5% (33.38 in 2009). Cases were reported in all the departments (even insular areas). As giardiasis is neglected in many countries, surveillance is not regularly undertaken. Despite its limitations, this study is the first attempt to provide estimates of national giardiasis incidence with consistent findings regarding affected age groups and geographical distribution.

Genotoxicity biomarkers for monitoring occupational exposure to antineoplastic drugs.
Rodríguez-Montero HM, Argote-Pelegrino E, Díaz-Curbelo A, Cuétara-Lugo EB. J Pharm Pharmacognosy Res. 2016 May 8;4(3):122–33.

Context The Institute of Oncology and Radiobiology (INOR) is the leading institution for the diagnosis, treatment and follow-up of cancer in Cuba. The main methods used in cancer treatment are surgery, radiotherapy and chemotherapy. The last one involves the handling of hazardous substances, such as cytostatics, which implies a health risk to persons occupationally exposed to it. There are two sites where a considerable among of cytostatic is handled (Ambulatory Chemotherapy Room (ACR) and the Central Unit of Cytostatic Mixture Preparation (CUCM)). Genotoxicity biomarkers of exposure and effects have been widely used to detect occupational environment hazards. Aims To evaluate genotoxicity biomarkers indicative of exposure and effects to cytostatics. Methods In this study were tested samples taken from the surfaces of biological safety cabinets located in the Central Unit of Cytostatic Mixture using SOS – Chromotest. We also evaluated samples of oral mucosa exfoliated cells from exposed and control subjects, by micronucleus test. Results All subjects were exposed and subjects who administered the mixes in the institution had an increase of DNA damage in comparisonwith the pharmaceutical staff that prepared it and wear the primary protection barriers properly. Conclusions These results underline the efficiency of genotoxicological biomarkers in detecting the exposure levels and the deleterious effect of cytostatics on occupationally exposed personal.

Healing enhancement of diabetic wounds by locally infiltrated epidermal growth factor is associated with systemic oxidative stress reduction.
Ojalvo AG, Acosta JB, Marí YM, Mayola MF, Pérez CV, Gutiérrez WS, et al. Int Wound J. 2016 Mar 22. DOI: 10.1111/iwj.12592. [Epub ahead of print]

The diabetic foot ulcer (DFU) is the leading cause of lower extremity amputation worldwide and is directly associated with comorbidity, disability and mortality. Oxidative stress mechanisms have been implicated in the pathogenesis of these wounds. Intra-lesional infiltration of epidermal growth factor has emerged as a potential therapeutic alternative to allow for physiological benefit while avoiding the proteolytic environment at the centre of the wound. The aim of this study was to characterise the response of patients with DFUs to epidermal growth factor treatment in terms of redox status markers. Experimental groups included patients with DFUs before and 3-4 weeks after starting treatment with epidermal growth factor; compensated and non-compensated diabetic patients without ulcers; and age-matched non-diabetic subjects. Evaluations comprised serum levels of oxidative stress and antioxidant reserve markers. Patients with DFUs exhibited the most disheveled biochemical profile, with elevated oxidative stress and low antioxidant reserves, with respect to non-ulcerated diabetic patients and to non-diabetic subjects. Epidermal growth factor intra-lesional administration was associated with a significant recovery of oxidative stress and antioxidant reserve markers. Altogether, our results indicate that epidermal growth factor intra-ulcer therapy contributes to restore systemic redox balance in patients with DFUs.

Incremental cost of implementing residual insecticide treatment with delthametrine on top of intensive routine Aedes aegypti control.
Baly A, González K, Cabrera P, Popa JC, Toledo ME, Hernández C, et al. Trop Med Int Health. 2016 Mar 21. DOI: 10.1111/tmi.12693. [Epub ahead of print]

Objective Information on the cost of implementing residual insecticide treatment (RIT) for Aedes control is scarce. We evaluated the incremental cost on top of intensive conventional routine activities of the Aedes control programme (ACP) in the city of Santiago de Cuba, Cuba. Methods We conducted the cost analysis study in 2011-2012, from the perspective of the ACP. Data sources were bookkeeping records, activity registers of the Provincial ACP Centre and the accounts of an RIT implementation study in 21 clusters of on average four house blocks comprising 5180 premises. Results The annual cost of the routine ACP activities was 19.66 US$ per household. RIT applications in rounds at 4-month intervals covering, on average, 97.2% and using 8.5 g of delthametrine annually per household, cost 3.06 US$ per household per year. Delthametrine comprised 66.5% of this cost; the additional cost for deploying RIT comprised 15.6% of the total ACP routine cost and 27% of the cost related to routine adult stage Aedes control. Conclusions The incremental cost of implementing RIT is high. It should be weighed against the incremental effect on the burden caused by the array of pathogens transmitted by Aedes. The cost could be reduced if the insecticide became cheaper, by limiting the number of yearly applications or by targeting transmission hot spots.

Indirect and competitive enzyme-linked immunosorbent assays for monitoring the humoral response against human VEGF.
Sánchez Ramírez J, Morera Díaz Y, Musacchio Lasa A, Bequet-Romero M, Muñoz Pozo Y, Pérez Sánchez L, et al. J Immunoassay Immunochem. 2016 May 4. [Epub ahead of print]

CIGB-247, a VEGF-based vaccine, was studied in a clinical trial. This advance demands the refinement of the methodologies for assessment of vaccine immune responses. This study aimed to improve the performance of ELISAs for detecting IgG antibodies against human VEGF and the blocking activity of the serum to inhibit the VEGF/VEGFR2 interaction. The best experimental conditions were established through the evaluation of several blocking buffers, immobilization surfaces and plate suppliers using human sera as test samples. As a result, two controlled ELISAs were used in testing of elicited immune response against VEGF in patients immunized with CIGB-247.

lQEEG and LORETA in teenagers with conduct disorder and psychopathic traits.
Calzada-Reyes A, Alvarez-Amador A, Galán-García L, Valdés-Sosa M. Clin CEEG Neurosci. 2016 Jun 7. pii: 1550059416645712. [Epub ahead of print]

Background Few studies have investigated the impact of the psychopathic traits on the EEG of teenagers with conduct disorder (CD). To date, there is no other research studying low-resolution brain electromagnetic tomography (LORETA) technique using quantitative EEG (QEEG) analysis in adolescents with CD and psychopathic traits. Objective To find electrophysiological differences specifically related to the psychopathic traits. The current investigation compares the QEEG and the current source density measures between adolescents with CD and psychopathic traits and adolescents with CD without psychopathic traits. Methods The resting EEG activity and LORETA for the EEG fast spectral bands were evaluated in 42 teenagers with CD, 25 with and 17 without psychopathic traits according to the Antisocial Process Screening Device. All adolescents were assessed using the DSM-IV-TR criteria. The EEG visual inspection characteristics and the use of frequency domain quantitative analysis techniques (narrow band spectral parameters) are described. Results QEEG analysis showed a pattern of beta activity excess on the bilateral frontal-temporal regions and decreases of alpha band power on the left central-temporal and right frontal-central-temporal regions in the psychopathic traits group. Current source density calculated at 17.18 Hz showed an increase within fronto-temporo-striatal regions in the psychopathic relative to the nonpsychopathic traits group. Conclusions These findings indicate that QEEG analysis and techniques of source localization may reveal differences in brain electrical activity among teenagers with CD and psychopathic traits, which was not obvious to visual inspection. Taken together, these results suggest that abnormalities in a fronto-temporo-striatal network play a relevant role in the neurobiological basis of psychopathic behavior.

Molecular detection and characterization of Anaplasma platys in dogs and ticks in Cuba.
Silva CB, Santos HA, Navarrete MG, Ribeiro CC, González BC, Zaldivar MF, et al. Ticks Tick Borne Dis. 2016 Apr 22. pii: S1877-959X(16)30068-1. DOI: 10.1016/j.ttbdis.2016.04.012. [Epub ahead of print]

Canine cyclic thrombocytopenia, an infectious disease caused by Anaplasma platys is a worldwide dog health problem. This study aimed to detect and characterize A. platys deoxyribonucleic acid (DNA) in dogs and ticks from Cuba using molecular methods. The study was conducted in four cities of Cuba (Habana del Este, Boyeros, Cotorro and San José de las Lajas). Blood samples were collected from 100 dogs in these cities. The animals were inspected for the detection of tick infestation and specimens were collected. Genomic DNA was extracted from dog blood and ticks using a commercial kit. Genomic DNA samples from blood and ticks were tested by a nested polymerase chain reaction (nPCR) to amplify 678 base pairs (bp) from the 16S ribosomal DNA (rDNA) of A. platys. Positive samples in nPCR were also subjected to PCR to amplify a fragment of 580bp from the citrate synthase (gltA) gene and the products were sequenced. Only Rhipicephalus sanguineus sensu lato (s.l.) was found on dogs, and 10.20% (n=5/49) of these ticks plus sixteen percent (16.0%, n=16/100) of dogs were considered positive for A. platys by nPCR targeting the 16S rDNA gene. All analyzed gltA and 16S rDNA sequences showed a 99-100% identity with sequences of A. platys reported in around the world. Phylogenetic analysis showed two defined clusters for the 16S rDNA gene and three defined clusters for the gltA gene. Based on the gltA gene, the deduced amino acid sequence showed two mutations at positions 88 and 168 compared with the sequence DQ525687 (GenBank ID from Italian sample), used as a reference in the alignment. A preliminary study on the epidemiological aspects associated with infection by A. platys showed no statistical association with the variables studied (p>0.05). This is the first evidence of the presence of A. platys in dogs and ticks in Cuba. Further studies are needed to evaluate the epidemiological aspects of A. platys infection in Cuban dogs.

Multicenter study highlighting clinical relevance of new high-throughput methodologies in molecular epidemiology of Pneumocystis jirovecii pneumonia.
Esteves F, de Sousa B, Calderón EJ, Huang L, Badura R, Maltez F, et al. Clin Microbiol Infect. 2016 Mar 25. pii: S1198-743X(16)30028-3. DOI: 10.1016/j.cmi.2016.03.013.

Pneumocystis jirovecii causes severe interstitial pneumonia (PcP) in immunosuppressed patients. This multicenter study assessed the distribution frequencies of epidemiologically relevant genetic markers of P. jirovecii in different geographic populations from Portugal, the USA, Spain, Cuba and Mozambique, and the relationship between the molecular data and the geographical and clinical information, based on a multifactorial approach. The high-throughput typing strategy for P. jirovecii characterization consisted of DNA pooling using quantitative real-time PCR (qPCR) followed by multiplex-PCR/single base extension (MPCR/SBE). The frequencies of relevant P. jirovecii single nucleotide polymorphisms (SNP) (mt85, SOD110, SOD215, DHFR312, DHPS165 and DHPS171) encoded at four loci were estimated in 10 DNA pooled samples representing a total of 182 individual samples. Putative multilocus genotypes (MLG) of P. jirovecii showed to be clustered due to the geographic differences but also dependent on clinical characteristics of the populations studied. The haplotype DHFR312T/SOD110C/SOD215T was associated with severe AIDS-related PcP and high P. jirovecii burdens. The frequencies of this genetic variant of P. jirovecii were significantly higher in patients with AIDS-related PcP from Portugal and the USA than in the colonized patients from Portugal, Spain, Cuba or Mozambique, highlighting the importance of this haplotype, apparently associated with the severity of the disease. Patients from the USA and Mozambique showedhigher rates of DHPS mutants, which may suggest the circulation of P. jirovecii organisms potentially related with TMP-SMX resistance in those geographical regions. This report assessed the worldwide distribution of P. jirovecii haplotypes and their epidemiological impact in distinct geographic and clinical populations.

Phylogenetic analysis of the Trypanosoma genus based on the heat-shock protein 70 gene.
Fraga J, Fernández-Calienes A, Montalvo AM, Maes I, Deborggraeve S, Büscher P, et al. Infect Genet Evol. 2016 May 13;43:165–72. DOI: 10.1016/j.meegid.2016.05.016. [Epub ahead of print]

Trypanosome evolution was so far essentially studied on the basis of phylogenetic analyses of small subunit ribosomal RNA (SSU-rRNA) and glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) genes. We used for the first time the 70kDa heat-shock protein gene (hsp70) to investigate the phylogenetic relationships among 11 Trypanosoma species on the basis of 1380 nucleotides from 76 sequences corresponding to 65 strains. We also constructed a phylogeny based on combined datasets of SSU-rDNA, gGAPDH and hsp70 sequences. The obtained clusters can be correlated with the sections and subgenus classifications of mammal-infecting trypanosomes except for Trypanosoma theileri and Trypanosoma rangeli. Our analysis supports the classification of Trypanosoma species into clades rather than in sections and subgenera, some of which being polyphyletic. Nine clades were recognized: Trypanosoma carassi, Trypanosoma congolense, Trypanosoma cruzi, Trypanosoma grayi, Trypanosoma lewisi, T. rangeli, T. theileri, Trypanosoma vivax and Trypanozoon. These results are consistent with existing knowledge of the genus’ phylogeny. Within the T. cruzi clade, three groups of T. cruzi discrete typing units could be clearly distinguished, corresponding to TcI, TcIII, and TcII+V+VI, while support for TcIV was lacking. Phylogenetic analyses based on hsp70 demonstrated that this molecular marker can be applied for discriminating most of the Trypanosoma species and clades.

Satisfaction with Life Scale (SLS-6): First validation study in Parkinson’s disease population.
Ambrosio L, Portillo MC, Rodríguez-Blazquez C, Martínez-Castrillo JC, Rodríguez-Violante M, Serrano-Dueñas M, et al. Parkinsonism Relat Disord. 2016 Apr;25:52–7.
DOI: 10.1016/j.parkreldis.2016.02.012. Epub 2016 Feb 15.

Introduction To explore the psychometric attributes of a new Satisfaction with Life Scale (SLS-6) in a wide Spanish-speaking population with Parkinson’s disease (PD). Methods This was an international, cross-sectional study. Several rater-based and patient-reported outcomes measures for evaluation of PD (e.g., Scales for Outcomes in Parkinson’s Disease-Motor) and other constructs (e.g., Duke-UNC Functional Social Support Questionnaire, Scale for Living with Chronic Illness) were applied together with the SLS-6. Acceptability, scaling assumptions, reliability, precision, and construct validity were tested. Results The study included 324 patients from five countries, with age (mean ± standard deviation) 66.67 ± 10.68 years. None of the SLS-6 items had missing values and all acceptability parameters fulfilled the standard criteria. Scaling assumptions allowed the calculation of a summary index from items 2 to 6, complementary to the global evaluation (item 1). For these five items, Cronbach’s alpha was 0.85; the corrected item-total correlation 0.53-0.73; inter-item correlation, 0.45-0.70, with an item homogeneity index of 0.55. The standard error of measurement, based on Cronbach’s alpha for a single observation, was 3.48. SLS-6 correlations were moderate to strong (rs ≥ 0.35) with the patient-reported outcomes and weak to moderate with the rater-based assessments used in the study. The SLS-6 total score was significantly different according to PD severity levels established according to Hoehn and Yahr staging, Clinical Impression of Severity Index, and Patient-Based Global Impression of Severity scale. Conclusion The results suggest that SLS-6 is an easy, feasible, acceptable, consistent, precise and valid measure to evaluate satisfaction with life in PD patients.

Selection of phage-displayed human antibody fragments specific for CD1b presenting the Mycobacterium tuberculosis glycolipid Ac2SGL.
Camacho F, Sarmiento ME, Reyes F, Kim L, Huggett J, Lepore M, et al. Int J Mycobacteriol. 2016 Jun;5(2):120–7.
DOI: 10.1016/j.ijmyco.2015.12.002. Epub 2016 Feb 17.

Objective/background The development of new tools capable of targeting Mycobacterium tuberculosis (Mtb)-infected cells has potential applications in diagnosis, treatment, and prevention of tuberculosis. In Mtb-infected cells, CD1b molecules present Mtb lipids to the immune system (Mtb lipid–CD1b complexes). Because of the lack of CD1b polymorphism, specific Mtb lipid–CD1b complexes could be considered as universal Mtb infection markers. 2-Stearoyl-3-hydroxyphthioceranoyl-2′-sulfate-α-α′-d-trehalose (Ac2SGL) is specific for Mtb, and is not present in other mycobacterial species. The CD1b–Ac2SGL complexes are expressed on the surface of human cells infected with Mtb. The aim of this study was to generate ligands capable of binding these CD1b–Ac2SGL complexes. Methods A synthetic human scFv phage antibody library was used to select phage-displayed antibody fragments that recognized CD1b–Ac2SGL using CD1b-transfected THP-1 cells loaded with Ac2SGL. Results One clone, D11—a single, light-variable domain (kappa) antibody (dAbκ11)—showed high relative binding to the Ac2SGL–CD1b complex. Conclusion A ligand recognizing the Ac2SGL–CD1b complex was obtained, which is a potential candidate to be further tested for diagnostic and therapeutic applications.

PT057 The hypertension in Cuba. Magnitude of the problem and control.
De la Noval DRJ, Dueñas A, Armas N, Acosta M. Global Heart. 2016 Jun;11(2 Suppl):e136.

According to the World Health Organization (WHO), cardiovascular diseases are a major cause of morbidity and mortality in adults in industrialized countries and developing.It is estimated that there are over 1 500 million worldwide hypertensive (blood pressure ≥140 / 90 mm Hg) and this figure is expected to increase more in the coming years. This disease affects approximately 30% of adults, so together with obesity classified by the WHO as pandemics of the century.

Therapeutic potential of the novel hybrid moleculeJM-20 against focal cortical ischemia in rats.
Núñez Figueredo Y, Ramírez Sánchez J, Hansel G, Pardo-Andreu GL, Merino N, Aparicio G, et al. J Pharm Pharmacognosy Res. 2016 Jun 7;4(4):153–8.

Context Despite the great mortality and morbidity of stroke, treatment options remain limited. We previously showed that JM-20, a novel synthetic molecule, possessed a strong neuroprotective effect in rats subjected to transient middle cerebral artery occlusion. However, to verify the robustness of the pre-clinical europrotective effects of JM-20 to get good prognosis in the translation to the clinic, it is necessary to use other experimental models of brain ischemia. Aims To evaluate the neuroprotective effects of JM-20 following the onset of permanent focal cerebral ischemia induced in rats by thermocoagulation of blood into pial blood vessels of cerebral cortices. Methods Ischemic lesion was induced by thermocoagulation of blood into pial blood vessels of primary motor and somatosensory cortices. Behavioral performance was evaluated by the cylinder testing for a period of 2, 3 and 7 days after surgery, and was followed by histopathological study in brain cortex stained with hematoxylineosin. Results Ischemic injury resulted in impaired function of the forelimb evidenced by high asymmetry punctuation, and caused histopathological alterations indicative of tissue damage at cerebral cortex. JM-20 treatment (4 and 8 mg/kg) significantly decreased asymmetry scores and histological alterations with a marked preservation of cortical neurons. Conclusions The effects of permanent brain ischemia were strongly attenuated by JM-20 administration, which expands and improves the current preclinical data of JM-20 as neuroprotector against cerebral ischemia, and strongly support the examination of its translation to the clinic to treat acute ischemic stroke.

The use of a three-dimensional endoscope for different skull base tumors: results of a preliminary extended endonasal surgical series.
Catapano G, de Notaris M, Di Maria D, Fernandez LA, Di Nuzzo G, Seneca V, et al. Acta Neurochir (Wien). 2016 Jun 8. [Epub ahead of print]

Background The evolution of skull base surgery over the past decade has been influenced by advancement in visualization technology. Recently, as a result of such improvements, three-dimensional (3-D) scopes have been widely used during endoscopic endonasal approaches. In the present study, we describe the use of 3-D stereoscopic endoscope for the treatment of a variety of skull base lesions. Methods From January 2010 to June 2015, a 3-D endoscopic endonasal approach (4 and 4.9 mm, 0°, and 30° rigid endoscopes) was performed in 70 patients with the following lesions: 42 large extrasellar pituitary macroadenomas, seven tuberculum sellae meningiomas, seven clivus chordomas, five craniopharyngiomas, three fibrous dysplasia of the clivus, three sinonasal malignancies, one orbital lymphangioma, one trigeminal neurinoma, one primary suprasellar lymphoma. Results Total tumor removal was obtained in 50 patients (71.4 %) while in 14 (20 %), subtotal removal was possible in six (8.6 %) only partial removal was achieved. Overall complications included diabetes insipidus in eight patients (11.4 %), hypopituitarism in seven patients (10 %), CSF leak in five patients (7.1 %), cranial nerve injury in two patients (2.8 %), panhypopituitarism in two patients (2.8 %), meningitis in one (1.4 %) and one postoperative central retinal artery occlusion (1.4 %). There was no mortality in the series. The mean follow-up time was 39 months (range, 6–72 months). Conclusions In our experience, the 3-D endoscope represents a critical development in visualization, thus enabling improved hand–eye coordination and depth perception, which are mandatory for the management of complex intradural neurovascular structures during tumor removal surgery.

Timing of influenza epidemics and vaccines in the American tropics, 2002-2008, 2011-2014.
Durand LO, Cheng PY, Palekar R, Clara W, Jara J, Cerpa M, et al. Influenza Other Respir Viruses. 2016 May;10(3):170–5. DOI: 10.1111/irv.12371. Epub 2016 Feb 8.

Background Influenza-associated illness results in increased morbidity and mortality in the Americas. These effects can be mitigated with an appropriately chosen and timed influenza vaccination campaign. To provide guidance in choosing the most suitable vaccine formulation and timing of administration, it is necessary to understand the timing of influenza seasonal epidemics. Objectives Our main objective was to determine whether influenza occurs in seasonal patterns in the American tropics and when these patterns occurred. Methods Publicly available, monthly seasonal influenza data from the Pan American Health Organization and WHO, from countries in the American tropics, were obtained during 2002-2008 and 2011-2014 (excluding unseasonal pandemic activity during 2009-2010). For each country, we calculated the monthly proportion of samples that tested positive for influenza. We applied the monthly proportion data to a logistic regression model for each country. Results We analyzed 2002-2008 and 2011-2014 influenza surveillance data from the American tropics and identified 13 (81%) of 16 countries with influenza epidemics that, on average, started during May and lasted 4 months. Conclusions The majority of countries in the American tropics have seasonal epidemics that start in May. Officials in these countries should consider the impact of vaccinating persons during April with the Southern Hemisphere formulation.

Unique human immune signature of Ebola virus disease in Guinea.
Ruibal P, Oestereich L, Lüdtke A, Becker-Ziaja B, Wozniak DM, Kerber R, et al. Nature. 2016 May 5;533(7601):100–4.
DOI: 10.1038/nature17949.

Despite the magnitude of the Ebola virus disease (EVD) outbreak in West Africa, there is still a fundamental lack of knowledge about the pathophysiology of EVD. In particular, very little is known about human immune responses to Ebola virus. Here we evaluate the physiology of the human T cell immune response in EVD patients at the time of admission to the Ebola Treatment Center in Guinea, and longitudinally until discharge or death. Through the use of multiparametric flow cytometry established by the European Mobile Laboratory in the field, we identify an immune signature that is unique in EVD fatalities. Fatal EVD was characterized by a high percentage of CD4(+) and CD8(+) T cells expressing the inhibitory molecules CTLA-4 and PD-1, which correlated with elevated inflammatory markers and high virus load. Conversely, surviving individuals showed significantly lower expression of CTLA-4 and PD-1 as well as lower inflammation, despite comparable overall T cell activation. Concomitant with virus clearance, survivors mounted a robust Ebola-virus-specific T cell response. Our findings suggest that dysregulation of the T cell response is a key component of EVD pathophysiology.

Validating non-invasive EEG source imaging using optimal electrode configurations on a representative rat head model.
Valdés-Hernández PA, Bae J, Song Y, Sumiyoshi A, Aubert-Vázquez E, Riera JJ. Brain Topogr. 2016 Mar 30. [Epub ahead of print]

The curtain of technical limitations impeding rat multichannel non-invasive electroencephalography (EEG) has risen. Given the importance of this preclinical model, development and validation of EEG source imaging (ESI) is essential. We investigate the validity of well-known human ESI methodologies in rats which individual tissue geometries have been approximated by those extracted from an MRI template, leading also to imprecision in electrode localizations. With the half and fifth sensitivity volumes we determine both the theoretical minimum electrode separation for non-redundant scalp EEG measurements and the electrode sensitivity resolution, which vary over the scalp because of the head geometry. According to our results, electrodes should be at least ~3 to 3.5 mm apart for an optimal configuration. The sensitivity resolution is generally worse for electrodes at the boundaries of the scalp measured region, though, by analogy with human montages, concentrates the sensitivity enough to localize sources. Cramér-Rao lower bounds of source localization errors indicate it is theoretically possible to achieve ESI accuracy at the level of anatomical structures, such as the stimulus-specific somatosensory areas, using the template. More validation for this approximation is provided through the comparison between the template and the individual lead field matrices, for several rats. Finally, using well-accepted inverse methods, we demonstrate that somatosensory ESI is not only expected but also allows exploring unknown phenomena related to global sensory integration. Inheriting the advantages and pitfalls of human ESI, rat ESI will boost the understanding of brain pathophysiological mechanisms and the evaluation of ESI methodologies, new pharmacological treatments and ESI-based biomarkers.