An ELISA for quantification of recombinant human EGF in production process samples, serum and urine. Leyva A, Santana H, Font M, Pérez B, Valdés R. Biologicals. 2017 Dec 12. pii : S1045-1056(17)30149-5. DOI: 10.1016/j.biologicals.2017.12.001. [Epub ahead of print]

A sandwich enzyme-linked immunosorbent assay for quantifying a recombinant human Epidermal Growth Factor (rhEGF) protein used in a vacunal preparation is described. The protein was detected with high specificity in a short incubation time at elevated temperature, the assay showing a linear range between 0.0625 and 1 ng/mL. According to the regression analysis for the dilutional linearity data, r2 = 0.9998, slope = 1.07 and intercept = 0.05 were obtained. The intra- and inter-assay coefficient of variation, ranged from 0.79 to 2.87% and 4.87-9.69% respectively demonstrating high reproducibility and precision. The ANOVA test used in the specificity/interference study revealed parallelism among curves (p > 0.1), which indicated lack of interference in the working range. Recovery obtained in accuracy test for three concentration levels varied between 89 and 111%; evidencing a reliable analytical assay to characterize the quality of the recombinant protein in the manufacturing process at large scale, and other biological matrixes as: urine and serum.

Anti-cancer activity of a new dihydropyridine derivative, VdiE-2N, in head and neck squamous cell carcinoma. Goto RN, Sobral LM, Sousa LO, García CB, Lopes NP, Marín-Prida J, et al. 2018 Jan 15;819:198–206.

This study aims to examine the effects of a new 1,4-dihydropyridine derivative, VdiE-2N, on cell signaling pathways and mitochondrial events in head and neck squamous cell carcinoma (HNSCC) cells, and on a mice model of xenograft tumor growth/cell proliferation. Four HNSCC cell lines (HN13, HN12, HN6, and CAL27), HEK293 cells (human embryonic kidney 293 cells), and human oral healthy mucosa fibroblasts (OHMF) were used for in vitro assessment of cell viability (resazurin assay) and invasion capacity (modified Boyden chamber assay), and mitochondrial membrane potential (JC-1 fluorescence assay), morphology (transmission electron microscopy), and number of mitochondria (MitoTracker® imaging). SET and pDRP1 proteins were analyzed by immunofluorescence, and proteins involved in cell death/survival pathways were analyzed by Western blotting. HN12 xenograft tumors were established in the flank of Balb/c nude mice, and their characteristics and sensitivity to VdiE-2N were determined by immunohistochemistry and histology. VdiE-2N decreased cell viability in HNSCC cells (IC50 = 9.56 and 22.45µM for HN13 and HN12 cells, respectively) more strongly than it decreased cell viability in OHMF and HEK293 cells (IC50 = 32.90 and > 50µM, respectively). In HN13 cells, VdiE-2N dissipated mitochondrial membrane potential and altered the mitochondria size, shape, and number in a concentration-dependent manner, as well as it induced apoptosis and reduced their invasion capacity. Treatment of mice bearing xenograft tumors with VdiE-2N significantly diminished proliferation of cancer cells. Therefore, VdiE-2N induces HNSCC cell death in vitro through mitochondria-mediated apoptotic pathways and dampens tumor growth in vivo, thus supporting a potential anti-cancer effect.

Antioxidant and Neuroprotective Effects of KM-34, A Novel Synthetic Catechol, Against Oxidative Stress-Induced Neurotoxicity. Nuñez-Figueredo Y, Ramírez-Sánchez J, Issac YA, Ochoa-Rodríguez E, Verdecia-Reyes Y, Delgado-Hernández R. Drug Res (Stuttg). 2017 Nov 3. DOI: 10.1055/s-0043-121220. [Epub ahead of print]

Free radicals are important mediators in a number of neurodegenerative diseases and molecules capable of scavenging reactive oxygen species (ROS) may be a feasible strategy for protecting neuronal cells. In this sense, polyphenols have been studied for their antioxidant effects, KM-34 (5-(3, 4-dydroxyl-benzylidene)-2, 2-dimethyl-1, 3-dioxane-4, 6-Dione) is a novel synthetic catechol with potential neuroprotective and antioxidant properties. We have assessed the antioxidant (as scavenging and iron-chelating compound) and neuroprotectant in vitro (in PC12 cell injury induced by H2O2, glutamate or FeSO4/AA) of KM-34. KM-34 was found to be a potent antioxidant, as shown by (i) inhibition of iron induced-brain lipid peroxidation, (ii) inhibition of 2-deoxyribose degradation, (iii) inhibition of superoxide radicals generation (IC50=11.04 μM) and (iv) inhibition of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical reduction (IC50=16.26  μM). The overall anti-oxidant action of KM-34 appears to be a combination of a direct reaction with free radicals and chelating the metal ions responsible for the production of ROS. Our work suggests that the antioxidant properties of KM-34 may provide future therapeutic approaches for neurodegenerative disorders.

A simple way for targeted delivery of an antibiotic: In vitro evaluation of a nanoclay-based composite. Valdés L, Pérez I, de Ménorval LC, Altshuler E, Fossum JO, Rivera A. PLoS One. 2017 Nov 17;12(11):e0187879. DOI: 10.1371/journal.pone.0187879. eCollection 2017.

The sodium-modified form of fluorohectorite nanoclay (NaFh) is introduced as a potential drug carrier, demonstrating its ability for the controlled release of the broad-spectrum antibiotic Ciprofloxacin through in vitro tests. The new clay-drug composite is designed to target the local infections in the large intestine, where it delivers most of the incorporated drug thanks to its pH-sensitive behavior. The composite has been conceived to avoid the use of coating technology and to decrease the side-effects commonly associated to the burst-release of the ciprofloxacin at the stomach level. NaFh was obtained from lithium-fluorohectorite by ion exchange, and its lack of toxicity was demonstrated by in vivo studies. Ciprofloxacin hydrochloride (Cipro) was encapsulated into the clay at different values of the pH, drug initial concentration, temperature and time. Systematic studies by X-ray diffraction (XRD), infrared and visible spectrophotometry (FT-IR and UV-vis), and thermal analysis (TGA) indicated that the NaFh host exhibits a high encapsulation efficiency for Cipro, which reaches a 90% of the initial Cipro in solution at 65 ℃, with initial concentration of drug in solution of 1.36 x 10-2 mol L-1 at acid pH. XRD revealed that a true intercalation of Cipro takes place between clay layers. TG showed an increased thermal stability of the drug when intercalated into the clay, as compared to the “free” Cipro. IR suggested a strong clay-Cipro interaction via ketone group, as well as the establishment of hydrogen bonds between the two materials. In vitro drug release tests revealed that NaFh is a potentially efficient carrier to deliver Cipro in the large intestine, where the release process is mediated by more than just one mechanism.

Barriers to Global Action Plan for the Prevention and Control of Non-communicable Diseases: Proposal Modifications to the Voluntary Targets. Morales Salinas A, Kones R. J Prevent Med. 2018;3(1):1–5.

World Health Organization member states gathered to draft the Global Action Plan for the prevention and control of noncommunicable diseases (NCDs). A global overarching target of a 25% reduction of premature mortality (between ages 30 and 70) from the four major NCDs (cardiovascular diseases, cancer, diabetes or chronic respiratory diseases) by 2025 was adopted. The primary aim of this mini-review is to identify the main barriers for reaching this plan using pertinent available evidence. The secondary aim is to reexamine the data with the dual purposes of amplifying beneficial effects of the voluntary global targets, and expanding the population to which they apply. This paper has identified five interrelated barriers: I) extremely low prevalence of ideal cardiovascular health, II) Gaps in affordability of essential medications, III) Gaps in health systems, IV) Low adherence to medications and V) Gaps in voluntary global targets. Conclusions: Modifications in the voluntary global targets are being proposed to produce greater effectiveness and extend reach. To this end, more ambitious risk factor targets for a larger proportion of the population and major efforts to strengthen healthcare systems are urgently needed.

Concentration Endurance Test (d2): Normative data for Spanish-speaking pediatric population. Rivera D, Salinas C, Ramos-Usuga D, Delgado-Mejía ID, Vasallo Key Y, Hernández Agurcia GP. NeuroRehabilitation. 2017 Oct 8. DOI 10.3233/NRE-172248. [Epub ahead of print]

Objective To generate normative data for the Concentration Endurance Test (d2) test in Spanish-speaking pediatric populations. Method The sample consisted of 4,373 healthy children from nine countries in Latin America (Chile, Cuba, Ecuador, Guatemala, Honduras, Mexico, Paraguay, Peru, and Puerto Rico) and Spain. Each participant was administered the d2 test as part of a larger neuropsychological battery. The Total number of items processed (TN), Total number of correct responses (CR), Total performance (TP), and Concentration performance (CP) scores were normed using multiple linear regressions and standard deviations of residual values. Age, age2, sex, and mean level of parental education (MLPE) were included as predictors in the analyses. Results The final multiple linear regression models showed main effects for age on all scores, such that scores increased linearly as a function of age. TN scores were affected by age2 for Guatemala and Puerto Rico respectively; CR scores were affected by age2 for Mexico; TP scores were affected by age2 for Chile, Mexico, Puerto Rico, and Spain; and CP scores for Mexico and Spain. Models indicated that children whose parents had a MLPE >12 years of education obtained higher scores compared to children whose parents had a MLPE≤12 years for Mexico, and Spain in all scores, and Puerto Rico for TN, CR, and TP, and Guatemala, and Paraguay for CP scores. Sex did not affect the scores for any country. Conclusions This is the largest Spanish-speaking pediatric normative study in the world, and it will allow neuropsychologists from these countries to have a more accurate approach to interpret the d2 test in pediatric populations.

Effectiveness of a Computer-Based Training Program of Attention and Memory in Patients with Acquired Brain Damage. Fernández E, Bergado Rosado JA, Rodríguez Pérez D, Salazar Santana S, Torres Aguilar M, Bringas ML. Behav Sci (Basel). 2017 Dec 30;8(1). pii : E4.

Many training programs have been designed using modern software to restore the impaired cognitive functions in patients with acquired brain damage (ABD). The objective of this study was to evaluate the effectiveness of a computer-based training program of attention and memory in patients with ABD, using a two-armed parallel group design, where the experimental group (n = 50) received cognitive stimulation using RehaCom software, and the control group (n = 30) received the standard cognitive stimulation (non-computerized) for eight weeks. In order to assess the possible cognitive changes after the treatment, a post-pre experimental design was employed using the following neuropsychological tests: Wechsler Memory Scale (WMS) and Trail Making test A and B. The effectiveness of the training procedure was statistically significant (p < 0.05) when it established the comparison between the performance in these scales, before and after the training period, in each patient and between the two groups. The training group had statistically significant (p < 0.001) changes in focused attention (Trail A), two subtests (digit span and logical memory), and the overall score of WMS. Finally, we discuss the advantages of computerized training rehabilitation and further directions of this line of work.

Electrocardiographic Markers of Appropriate Implantable Cardioverter-Defibrillator Therapy in Young People with Congenital Heart Diseases. Benítez Ramos DB, Cabrera Ortega M, Castro Hevia J, Dorantes Sánchez M, Alemán Fernández AA, Castañeda Chirino O, et al. Pediatr Cardiol. 2017 Sep 4. DOI: 10.1007/s00246-017-1711-9. [Epub ahead of print]

Implantable cardioverter-defibrillators (ICDs) are increasingly utilized in patients with congenital heart disease (CHD). Prediction of the occurrence of shocks is important if improved patient selection is desired. The electrocardiogram (ECG) has been the first-line tool predicting the risk of sudden death, but data in CHD patients are lacking. We aim to evaluate the predictive value of electrocardiographic markers of appropriate therapy of ICD in young people with CHD. We conducted a prospective, longitudinal study, in twenty-six CHD patients (mean age 24.7 ± 5.3 years) who underwent first ICD implantation. Forty-two age- and diagnosis-matched controls were recruited. Twelve-lead ECG and 24 h Holter analysis were performed during a mean follow-up of 38.9 months. Data included heart rate, heart rate variability, QRS duration (QRSd), QTc interval and its dispersion, Tpeak-Tend (Tp-Te) interval and its dispersion, presence of fragmented QRS (fQRS), T wave alternans, atrial arrhythmias, and non-sustained ventricular tachycardia. Implant indication was primary prevention in ten cases (38.5%) and secondary prevention in 16 (61.5%). Overall, 17 subjects (65.3%) received at least one appropriate and effective ICD discharge. fQRS was present in 64.7% of cases with ICD therapy compared with patients without events or controls (p < 0.0001). Tp-e and Tp-e dispersion were significantly prolonged in patients with recurrences (113.5 and 37.2 ms) versus patients without ICD discharge (89.6 and 24.1 ms) or controls (72.4 and 19.3 ms) (p < 0.0001 and p < 0.0001, respectively). On univariate Cox regression analysis QRSd (hazard ratio: 1.19 per ms, p = 0.003), QTc dispersion (hazard ratio: 1.57 per ms, p = 0.002), fQRS (hazard ratio: 3.58 p < 0.0001), Tp-e (hazard ratio: 2.27 per ms, p < 0.0001), and Tp-e dispersion (hazard ratio: 4.15 per ms, p < 0.0001), emerged as strong predictors of outcome. On multivariate Cox analysis fQRS, Tp-e and Tp-e dispersion remained in the model. The presence of fQRS, and both Tp-e and Tp-e dispersion are useful ECG tools in daily clinical practice to identify CHD patients at risk for appropriate ICD therapy.

Fast Gaussian Naïve Bayes for searchlight classification analysis. Ontivero-Ortega M, Lage-Castellanos A, Valente G, Goebel R, Valdés-Sosa M. Neuroimage. 2017 Sep 4. pii: S1053-8119(17)30737-1. DOI: 10.1016/j.neuroimage.2017.09.001. [Epub ahead of print]

The searchlight technique is a variant of multivariate pattern analysis (MVPA) that examines neural activity across large sets of small regions, exhaustively covering the whole brain. This usually involves application of classifier algorithms across all searchlights, which entails large computational costs especially when testing the statistical significance of the accuracies with permutation methods. In this article, a new implementation of the Gaussian Naive Bayes classifier is presented (henceforth massive-GNB). This approach allows classification in all searchlights simultaneously, and is faster than previously published searchlight GNB implementations, as well as other more complex classifiers including support vector machines (SVM). To ensure that the gain in speed for GNB would be useful in searchlight analysis, we compared the accuracies of massive-GNB and SVM in detecting the lateral occipital complex (LOC) in an fMRI localizer experiment (26 subjects). Moreover, this region as defined in a meta-analysis of many activation studies was used as a gold standard to compare error rates for both classifiers. In individual searchlights, SVM was somewhat more accurate than massive-GNB and more selective in detecting the meta-analytic LOC. However, with multiple comparison correction at the cluster-level the two classifiers performed equivalently. Thus for cluster-level analysis, massive-GNB produces an accuracy similar to more sophisticated classifiers but with a substantial gain in speed. Massive-GNB (available as a public Matlab toolbox) could facilitate the more widespread use of searchlight analysis.

Growth hormone releasing peptide-6 enhanced antibody titers against subunit antigens in mice (BALB/c), tilapia (Oreochromis niloticus) and African catfish (Clarias gariepinus). Martínez R, Hernández L, Gil L, Carpio Y, Morales A, Herrera F, et al. Vaccine. 2017 Oct 9;35(42):5722–8. DOI: 10.1016/j.vaccine.2017.07.060.

Modern subunit vaccines have excellent safety profiles and improved tolerability, but do not elicit strong immune responses without the addition of adjuvants. Developing a safe and affective adjuvant remains a challenge for peptide-based vaccine design. Growth Hormone Releasing Peptide-6 (GHRP-6) is one of the earliest-developed, synthetic, peptidyl growth hormone secretagogue receptor agonists. These compounds mimic the effect of the endogenous ligand, ghrelin. In the present study, we evaluated the ability of GHRP-6 to enhance the humoral immune response against co-injected antigens in mice, tilapia and African catfish. This peptide was able to increase the antigen-specific antibody response using heterologous proteins and peptides as antigens, which were also formulated in “water in oil” emulsions (Freund and Montanide). As long as we know there is no previous report describing any ghrelin analogous as molecular immunomodulator stimulating a humoral immune response. Further studies will be conducted to evaluate the functionality of this humoral immune response in challenge trials.

Heritability of saccadic eye movements in spinocerebellar ataxia type 2: insights into an endophenotype marker. Rodríguez-Labrada R, Vázquez-Mojena Y, Canales-Ochoa N, Medrano-Montero J, Velázquez-Pérez L. Cerebellum Ataxias. 2017 Dec 19;4:19.

Background Saccade slowing has been proposed as endophenotype marker in Spinocerebellar Ataxia type 2 (SCA2), nevertheless the heritability of this trait has not been properly demonstrated. Thus the present paper was aimed to assess the heritability of different saccadic parameters in SCA2. Methods Forty-eight SCA2 patients, 25 preclinical carriers and 24 non-SCA2 mutation carriers underwent electronystagmographical assessments of saccadic eye movements as well as neurological examination and ataxia scoring. Estimates of heritability based on the intraclass correlation coefficients were calculated for saccade velocity, accuracy and latency as well as for age at disease onset from 36, 17 and 15 sibling pairs of SCA2 patients, preclinical carriers and controls, respectively. Results Saccade velocity was significantly reduced in SCA2 patients and preclinical carriers, whereas decreased saccade accuracy and increased saccade latency were only observed in the patients cohort. Intraclass correlation coefficient for saccade velocity was highly significant in SCA2 patients, estimating a heritability around 94%, whereas for the age at ataxia onset this estimate was around 68%. Conclusions Electronystagmographical measure of saccade velocity showed higher familial aggregation between SCA2 patients leading the suitability of this disease feature as endophenotype marker, with potential usefulness for the search of modifier genes and neurobiological underpinnings of the disease and as outcome measure in future neuroprotective clinical trials.

KM-34, a Novel Antioxidant Compound, Protects against 6-Hydroxydopamine-Induced Mitochondrial Damage and Neurotoxicity. Fonseca-Fonseca LA, Núñez-Figueredo Y, Sánchez JR, Guerra MW, Ochoa-Rodríguez E, Verdecia-Reyes Y, et al. Neurotox Res. 2018 Jan 2.
DOI: 10.1007/s12640-017-9851-5. [Epub ahead of print]

The etiology of Parkinson’s disease is not completely understood and is believed to be multifactorial. Neuronal disorders associated to oxidative stress and mitochondrial dysfunction are widely considered major consequences. The aim of this study was to investigate the effect of the synthetic arylidenmalonate derivative 5-(3,4-dihydroxybenzylidene)-2,2-dimethyl-1,3-dioxane-4,6-dione (KM-34), in oxidative stress and mitochondrial dysfunction induced by 6-hydroxydopamine (6-OHDA). Pretreatment (2 h) with KM-34 (1 and 10 μM) markedly attenuated 6-OHDA-induced PC12 cell death in a concentration-dependent manner. KM-34 also inhibited H2O2 generation, mitochondrial swelling, and membrane potential dissipation after 6-OHDA-induced mitochondrial damage. In vivo, KM-34 treatment (1 and 2 mg/Kg) reduced percentage of asymmetry (cylinder test) and increased the vertical exploration (open field) with respect to untreated injured animals; KM-34 also reduced glial fibrillary acidic protein overexpression and increased tyrosine hydroxylase-positive cell number, both in substantia nigra pars compacta. These results demonstrate that KM-34 present biological effects associated to mitoprotection and neuroprotection in vitro, moreover, glial response and neuroprotection in SNpc in vivo. We suggest that KM-34 could be a putative neuroprotective agent for inhibiting the progressive neurodegenerative disease associated to oxidative stress and mitochondrial dysfunction.

Limits of aerobic metabolism in cancer cells. Fernández-de-Cossio-Díaz J, Vázquez A. Sci Rep. 2017 Oct 18;7(1):13488. DOI: 10.1038/s41598-017-14071-y.

Cancer cells exhibit high rates of glycolysis and glutaminolysis. Glycolysis can provide energy and glutaminolysis can provide carbon for anaplerosis and reductive carboxylation to citrate. However, all these metabolic requirements could be in principle satisfied from glucose. Here we investigate why cancer cells do not satisfy their metabolic demands using aerobic biosynthesis from glucose. Based on the typical composition of a mammalian cell we quantify the energy demand and the OxPhos burden of cell biosynthesis from glucose. Our calculation demonstrates that aerobic growth from glucose is feasible up to a minimum doubling time that is proportional to the OxPhos burden and inversely proportional to the mitochondria OxPhos capacity. To grow faster cancer cells must activate aerobic glycolysis for energy generation and uncouple NADH generation from biosynthesis. To uncouple biosynthesis from NADH generation cancer cells can synthesize lipids from carbon sources that do not produce NADH in their catabolism, including acetate and the amino acids glutamate, glutamine, phenylalanine and tyrosine. Finally, we show that cancer cell lines have an OxPhos capacity that is insufficient to support aerobic biosynthesis from glucose. We conclude that selection for high rate of biosynthesis implies a selection for aerobic glycolysis and uncoupling biosynthesis from NADH generation.

Newly developed Learning and Verbal Memory Test (TAMV-I): Normative data for Spanish-speaking pediatric population. Rivera D, Olabarrieta-Landa L, Rabago Barajas BV, Irías Escher MJ, Saracostti Schwartzman M, Ferrer-Cascales R, et al. NeuroRehabilitation. 2017 Oct 10. DOI: 10.3233/NRE-172249. [Epub ahead of print]

Objective To generate normative data for the Learning and Verbal Memory Test (TAMV-I) in Spanish-speaking pediatric populations. Method The sample consisted of 4,373 healthy children from nine countries in Latin America (Chile, Cuba, Ecuador, Guatemala, Honduras, Mexico, Paraguay, Peru, and Puerto Rico) and Spain. Each participant was administered the TAMV-I as part of a larger neuropsychological battery. Free recall, memory delay and recognition scores were normed using multiple linear regressions and standard deviations of residual values. Age, age2, sex, and mean level of parental education (MLPE) were included as predictors in the analyses. Results The final multiple linear regression models indicated main effects for age on all scores, such that scores increased linearly as a function of age. Age2 had a significant effect in all countries except Cuba, Paraguay, and Puerto Rico for free recall score; a significant effect for memory delay score in all countries except Cuba and Puerto Rico; and a significant effect for recognition score in in all countries except Guatemala, Honduras, and Puerto Rico. Models showed an effect for MLPE in Chile (free recall), Honduras (free recall), Mexico (free recall), Puerto Rico (free recall, memory delay and recognition), and Spain (free recall and memory delay). Sex affected free recall score for Cuba, Ecuador, Guatemala, Mexico, Paraguay, Peru, and Spain, memory delay score for all countries except Chile, Paraguay, and Puerto Rico, and recognition score for Ecuador, Mexico, Peru, and Spain, with girls scoring higher than boys. Conclusions This is the largest Spanish-speaking pediatric normative study in the world, and it will allow neuropsychologists from these countries to have a more accurate way to interpret the TAMV-I with pediatric populations.

Non-Clinical Models for Neurodegenerative Diseases: Therapeutic Approach and Drug Validation in Animal Models. Fernández CI, López J, Martínez L. Behav Sci (Basel). 2017 Dec 4;7(4):82.

In 2016, 19.8% of the Cuban population was aged 60 or over. As a result, age-associated degenerative diseases and other diseases have become priority targets from a prophylactic, diagnostic and therapeutic perspective. As a result, the Cuban biomedical scientific community has addressed its basic, preclinical and epidemiological research in order to rise up to the challenge. A firm step in this direction has been the international congress “State of the art in non-clinical models for neurodegenerative diseases” which has brought together preclinical and clinical researchers, technicians and regulatory staff members from different countries to review the state of the art in neurodegenerations, find unifying ideas, objectives and collaborations or partnership. The objective is to expose the perspectives of new biotechnological products from Cuba and other countries from the diagnostic, therapeutic and neuroprotective point of view. It is crucial, therefore, that the irreplaceable role of laboratory animals in achieving these objectives is understood but they must be used in rational, adequate and ethical manner. We expose the current development trends in this field, being of common interest to the work directed to the search for potential drugs, diagnostic tools and the promotion of changes in lifestyle as a preventive projection.

Overall Survival of Patients with Locally Advanced or Metastatic Esophageal Squamous Cell Carcinoma Treated with Nimotuzumab in the Real World. Saumell Y, Sánchez L, González S, Ortiz R, Medina E, Galán Y. Adv Ther. 2017 Nov 13. DOI: 10.1007/s12325-017-0631-7. [Epub ahead of print].

Introduction Despite improvements in surgical techniques and treatments introduced into clinical practice, the overall survival of patients with esophageal squamous cell carcinoma remains low. Several epidermal growth factor receptor inhibitors are being evaluated in the context of clinical trials, but there is little evidence of effectiveness in real-world conditions. This study aimed at assessing the effectiveness of nimotuzumab combined with onco-specific treatment in Cuban real-life patients with locally advanced or metastatic esophageal squamous cell carcinoma. Methods A comparative and retrospective effectiveness study was performed. The 93 patients treated with nimotuzumab were matched, with use of propensity score matching, with patients who received a diagnosis of locally advanced or metastatic squamous cell carcinoma of the esophagus in three Cuban provinces reported between 2011 and 2015 to the National Cancer Registry. The Kaplan-Meier method was used to estimate event-time distributions. Log-rank statistics were used for comparisons of overall survival between groups. A two-component mixture model assuming a Weibull distribution was fitted to assess the effect of nimotuzumab on short-term and long-term survival populations. Results There was an increase in median overall survival in patients treated with nimotuzumab (11.9 months versus 6.5 months without treatment) and an increase in the 1-year survival rate (54.0% versus 21.9% without treatment). The 2-year survival rates were 21.1% for patients treated with nimotuzumab and 0% in the untreated cohort. There were statistically significant differences in survival between groups treated and not treated with nimotuzumab, both in the short-term survival population (6.0 months vs 4.0 months, p = 0.009) and in the long-term survival population (18.0 months vs 11.0 months, p = 0.001). Conclusions Our study shows that nimotuzumab treatment concurrent with chemoradiotherapy increases the survival of real-world patients with locally advanced or metastatic esophageal squamous cell carcinoma. Further prospective studies are required to confirm the therapeutic effectiveness of nimotuzumab in esophageal cancer.

Pharmacokinetics comparison of two pegylated interferon alfa formulations in healthy volunteers. Costa MB, Picon BD, Sander GB, Cuni HN, Silva CV, Meireles RP, et al. BMC Pharmacol Toxicol. 2018 Jan 4;19(1):1.

Background Several countries have used pegylation technology to improve the pharmacokinetic properties of essential drugs. Recently, a novel interferon alfa-2b protein conjugated to four-branched 12 kDa polyethylene glycol molecules was developed jointly between Cuba and Brazil. The aim of this study was to compare the pharmacokinetic properties of BIP48 (pegylated interferon alfa-2b from Bio-Manguinhos/Fiocruz, Brazil) to those of PEGASYS® (commercially available pegylated interferon alfa-2a from Roche Pharmaceutical). Methods This phase I, single-centre, randomized, double-blind crossover trial enrolled 31 healthy male volunteers aged 19 to 35 who were allocated to two stages, either side of a 5-week wash-out period, with each arm lasting 14 consecutive days after subcutaneous administration of 180 μg of one formulation or the other (study or comparator). The main outcome variable was serum pegylated interferon concentrations in 15 samples collected during the course of the study and tested using an enzyme immunoassay. Results There were no differences between formulations in terms of magnitude or absorption parameters. Analysis of time parameters revealed that BIP48 remained in the body significantly longer than PEGASYS® (Tmax: 73 vs. 54 h [p = 0.0010]; MRT: 133 vs. 115 h [p = 0.0324]; ke: 0.011 vs. 0.013 h(-1) [p = 0.0153]; t1/2: 192 vs. 108 h [p = 0.0218]). Conclusion BIP48 showed the expected pharmacokinetic profile for a pegylated product with a branched molecular structure. Compared to PEGASYS®, the magnitude absorption was similar, but time parameters were consistent with slower elimination. Further studies should be conducted to evaluate the clinical implications of these findings. A phase II-III repeated-dose clinical trial is ongoing to study these findings in patients with chronic hepatitis C virus infection. Trial Registration This study is registered on the ClinicalTrials.gov platform (accession number NCT01889849 ). This trial was retrospectively registered in June 2013.

Reliability and Validity of Nonsymbolic and Symbolic Comparison Tasks in School-Aged Children. Castro D, Estévez N, Gómez D, Dartnell PR. Span J Psychol2017 Dec 4;20:E75.
DOI: 10.1017/sjp.2017.68.

Basic numerical processing has been regularly assessed using numerical nonsymbolic and symbolic comparison tasks. It has been assumed that these tasks index similar underlying processes. However, the evidence concerning the reliability and convergent validity across different versions of these tasks is inconclusive. We explored the reliability and convergent validity between two numerical comparison tasks (nonsymbolic vs. symbolic) in school-aged children. The relations between performance in both tasks and mental arithmetic were described and a developmental trajectories’ analysis was also conducted. The influence of verbal and visuospatial working memory processes and age was controlled for in the analyses. Results show significant reliability (p < .001) between Block 1 and 2 for nonsymbolic task (global adjusted RT (adjRT): r = .78, global efficiency measures (EMs): r = .74) and, for symbolic task (adjRT: r = .86, EMs: r = .86). Also, significant convergent validity between tasks (p < .001) for both adjRT (r = .71) and EMs (r = .70) were found after controlling for working memory and age. Finally, it was found the relationship between nonsymbolic and symbolic efficiencies varies across the sample’s age range. Overall, these findings suggest both tasks index the same underlying cognitive architecture and are appropriate to explore the Approximate Number System (ANS) characteristics. The evidence supports the central role of ANS in arithmetic efficiency and suggests there are differences across the age range assessed, concerning the extent to which efficiency in nonsymbolic and symbolic tasks reflects ANS acuity.

Rey- Osterrieth Complex Figure – copy and immediate recall (3 minutes): Normative data for Spanish-speaking pediatric populations. Arango-Lasprilla JC, Rivera D, Erlt MM, Muñoz Mancilla JM, García-Guerrero CE, Rodríguez-Irizarry W, et al. NeuroRehabilitation. 2017 Sep 5. DOI: 10.3233/NRE-172241. [Epub ahead of print]

Objective To generate normative data for the Rey- Osterrieth Complex Figure (ROCF) in Spanish-speaking pediatric populations. Method The sample consisted of 4,373 healthy children from nine countries in Latin America (Chile, Cuba, Ecuador, Guatemala, Honduras, Mexico, Paraguay, Peru, and Puerto Rico) and Spain. Each participant was administered the ROCF as part of a larger neuropsychological battery. The ROCF copy and immediate recall (3 minutes) scores were normed using multiple linear regressions and standard deviations of residual values. Age, age2, sex, and mean level of parental education (MLPE) were included as predictors in the analyses. Results The final multiple linear regression models showed main effect for age on copy and immediate recall scores, such that scores increased linearly as a function of age. Age2 affected ROCF copy scores for all countries, except Puerto Rico; and ROCF immediate recall scores for all countries, except Chile, Guatemala, Honduras, Paraguay, and Puerto Rico. Models indicated that children whose parents had a MLPE >12 years of education obtained higher scores compared to children whose parents had a MLPE≤12 years for Chile, Puerto Rico, and Spain in the ROCF copy, and Paraguay and Spain for the ROCF immediate recall. Sex affected ROCF copy and immediate recall scores for Chile and Puerto Rico with girls scoring higher than boys. Conclusions This is the largest Spanish speaking pediatric normative study in the world, and it will allow neuropsychologists from these countries to have a more accurate approach to interpret the ROCF Test in pediatric populations.

Risk factors and outcomes of severe acute respiratory failure requiring invasive mechanical ventilation in cancer patients: A retrospective cohort study. Martos-Benítez FD, Gutiérrez-Noyola A, Badal M, Dietrich NA. Med Intensiva. 2017 Sep 28. pii: S0210-5691(17)30226-7. DOI: 10.1016/j.medin.2017.08.004. [Epub ahead of print]

Objectives To determine the risk factors for severe acute respiratory failure requiring invasive mechanical ventilation (SARF-MV) and its effect upon clinical outcomes in critically ill cancer patients. Design A retrospective cohort study was carried out. Setting A 12-bed oncological intensive care unit (ICU) from January 2014 to December 2015. PATIENTSA total of 878 consecutive cancer patients were included. Patients with an ICU stay of ≤1 day were excluded. The final sample size was 691 patients. Interventions None. Variables Clinical variables at ICU admission were extracted from the medical records. The primary outcome was SARF-MV. We also measured ICU and hospital mortality, as well as length of stay. Results The SARF-MV rate was 15.8%. The multivariate analysis identified brain tumor (OR 14.54; 95%CI 3.86-54.77; p<0.0001), stage IV cancer (OR 3.47; 95%CI 1.26-9.54; p=0.016), sepsis upon admission (OR 2.28; 95%CI 1.14-4.56; p=0.020) and an APACHE II score≥20 points (OR 5.38; 95%CI 1.92-15.05; p=0.001) as being independently associated to SARF-MV. Compared with the patients without SARF-MV, those with SARF-MV had a prolonged length of ICU stay (p<0.0001), a lower ICU survival rate (p<0.0001) and a lower hospital survival rate (p<0.0001). Conclusions A number of clinical factors are related to SARF-MV. In this regard, SARF-MV is a powerful factor independently correlated to poor outcomes. Future studies should investigate means for preventing SARF-MV in critically ill cancer patients, which may have an impact upon outcomes.

Sepiolite as a New Nanocarrier for DNA Transfer into Mammalian Cells: Proof of Concept, Issues and Perspectives. Piétrement O, Castro-Smirnov FA, Le Cam E, Aranda P, Ruiz-Hitzky E, López BS. Chem Rec. 2017 Dec 29. DOI: 10.1002/tcr.201700078. [Epub ahead of print]

Sepiolite is a nanofibrous natural silicate that can be used as a nanocarrier for DNA transfer thanks to its strong interaction with DNA molecules and its ability to be naturally internalized into mammalian cells through both non-endocytic and endocytic pathways. Sepiolite, due to its ability to bind various biomolecules, could be a good candidate for use as a nanocarrier for the simultaneous vectorization of diverse biological molecules. In this paper, we review our recent work, issued from a starting collaboration with Prof. Ruiz-Hitzky, that includes diverse aspects on the characterization and main features of sepiolite/DNA nanohybrids, and we present an outlook for the further development of sepiolite for DNA transfer.

Sustained attention in school-age children with congenital hypothyroidism: Influence of episodes of overtreatment in the first three years of life. García Morales L, Rodríguez Arnao MD, Rodríguez Sánchez A, Dulín Íñiguez E, Álvarez González MA. Neurologia. 2017 Nov 18. pii: S0213-4853(17)30299-2. DOI: 10.1016/j.nrl.2017.08.003. [Epub ahead of print]. English, Spanish.

Introduction Children with congenital hypothyroidism (CH) are at risk of developing mild cognitive impairment despite normal overall intellectual performance. These deficits may be caused by disease-related and treatment-related factors. This study explores the impact of abnormal thyroid function during the first 3 years of life on attention performance at school age. Methods We included 49 children diagnosed with CH and receiving treatment for the condition: 14 boys (mean age 9.5±2.8 years) and 35 girls (9.6±2.6 years). The number of episodes of normal, under-, and overtreatment were estimated based on TSH levels during their first 3 years of life (at 12, 18, 24, 30, and 36 months). Children were assessed using a computerised version of a Sustained attention test. General linear models were calculated with the attention index as the dependent variable and sex, aetiology, and number of episodes of normal, under-, and overtreatment as independent variables. Results Higher numbers of episodes of overtreatment (low TSH level) were associated with poorer attention performance at school age (P=.005, r=-0.45). Conclusions Children with CH should be monitored closely during the first 3 years of life in order to prevent not only hypothyroidism but also any adverse effects of overtreatment that may affect attentional function at school age.

The additional benefit of residual spraying and insecticide-treated curtains for dengue control over current best practice in Cuba: Evaluation of disease incidence in a cluster randomized trial in a low burden setting with intensive routine control. Toledo ME, Vanlerberghe V, Rosales JP, Mirabal M, Cabrera P, Fonseca V, et al. PLoS Negl Trop Dis. 2017 Nov 8;11(11):e0006031. DOI: 10.1371/journal.pntd.0006031.

Background Aedes control interventions are considered the cornerstone of dengue control programmes, but there is scarce evidence on their effect on disease. We set-up a cluster randomized controlled trial in Santiago de Cuba to evaluate the entomological and epidemiological effectiveness of periodical intra- and peri-domiciliary residual insecticide (deltamethrin) treatment (RIT) and long lasting insecticide treated curtains (ITC). Methodology/Principal findings Sixty three clusters (around 250 households each) were randomly allocated to two intervention (RIT and ITC) and one control arm. Routine Aedes control activities (entomological surveillance, source reduction, selective adulticiding, health education) were applied in the whole study area. The outcome measures were clinical dengue case incidence and immature Aedes infestation. Effectiveness of tools was evaluated using a generalized linear regression model with a negative binomial link function. Despite significant reduction in Aedes indices (Rate Ratio (RR) 0.54 (95%CI 0.32-0.89) in the first month after RIT, the effect faded out over time and dengue incidence was not reduced. Overall, in this setting there was no protective effect of RIT or ITC over routine in the 17months intervention period, with for house index RR of 1.16 (95%CI 0.96-1.40) and 1.25 (95%CI 1.03-1.50) and for dengue incidence RR of 1.43 (95%CI 1.08-1.90) and 0.96 (95%CI 0.72-1.28) respectively. The monthly dengue incidence rate (IR) at cluster level was best explained by epidemic periods (Incidence Rate Ratio (IRR) 5.50 (95%CI 4.14-7.31)), the IR in bordering houseblocks (IRR 1.03 (95%CI 1.02-1.04)) and the IR pre-intervention (IRR 1.02 (95%CI 1.00-1.04)). Conclusions Adding RIT to an intensive routine Aedes control programme has a transient effect on the already moderate low entomological infestation levels, while ITC did not have any effect. For both interventions, we didn’t evidence impact on disease incidence. Further studies are needed to evaluate impact in settings with high Aedes infestation and arbovirus case load.

The anti-CD6 antibody itolizumab provides clinical benefit without lymphopenia in rheumatoid arthritis patients: results from a 6-month, open-label phase I clinical trial. Rodríguez PC, Prada DM, Moreno E, Aira LE, Molinero C, López AM, et al. Clin Exp Immunol. 2017 Sep 30. DOI: 10.1111/cei.13061. [Epub ahead of print]

Itolizumab is a humanized anti-CD6 monoclonal antibody (mAb) that has previously shown encouraging results, in terms of safety and positive clinical effects, in a 6-week monotherapy clinical trial conducted in RA patients. The current phase I study evaluated the safety and clinical response for a longer treatment of 12 itolizumab intravenous doses in subjects with active RA despite previous DMARD therapy. Twenty one subjects were enrolled in four dose groups (0.1, 0.2, 0.4 and 0.8 mg/kg). Efficacy endpoints including American College of Rheumatology (ACR)20, ACR50 and ACR70 response rates and Disease Activity Score in 28 joints (DAS28) were monitored at baseline and at specific time points over a 10-week follow-up period. Itolizumab was well tolerated up to the highest tested dose. No related serious adverse events were reported and most adverse events were mild. Remarkably, itolizumab treatment did not produce lymphopenia and, therefore, was not associated with infections. All patients achieved a clinical response (ACR20) at least once during the study. Eleven subjects (55%) achieved at least a 20% improvement in ACR just one week after the first itolizumab administration. The clinical response was observed from the beginning of the treatment and was sustained over 24 weeks. The efficacy profile of this 12-week treatment was similar to that of the previous study (6-week treatment). These results reinforce the safety profile of itolizumab and provide further evidence on the clinical benefit from the use of this anti-CD6 mAb in RA patients. This article is protected by copyright. All rights reserved.

The first report of cases of pet dogs with naturally occurring cancer treated with the antitumor peptide CIGB-552.Vallespi MG, Rodríguez JC, Seoane LC, Álvarez P, Santana H, Garay H, et al. Res Vet Sci. 2017 Oct;114:502–10. DOI: 10.1016/j.rvsc.2017.09.029. Epub 2017 Sep 29.

The absence of an effective therapy against human solid tumors has fostered the development of promising antineoplastic therapeutic candidates, as the CIGB-552 peptide. This synthetic peptide has shown to be effective in reducing tumor size and increasing the lifespan in tumor-bearing mice. Therefore, this work was aimed to explore the safety profile and preliminary assessment of antitumor activity of the CIGB-552 peptide therapeutic candidate in a small population of dogs (n=9) having malignant spontaneously-arising solid tumors. The peptide was administered by subcutaneous (s.c.) route, at three dosage levels (0.075, 0.15 and 0.3mg/kg). The results showed no dose-limiting toxicities in any dogs. The antitumor activity observed in dogs receiving CIGB-552 was associated with the reduction in the tumor volume. Given the antitumor effects of CIGB-552 as mediated by COMMD1 protein, which function is highly conserved among eukaryotic organisms, and the similarities of canine and human types of cancer with respect to tumor biology, it is likely that CIGB-552 could demonstrate comparable anti-cancer activity in human patients.

The immune profile induced is crucial to determine the effects of immunocastration over gonadal function, fertility, and GnRH-I expression. Siel D, Loaiza A, Vidal S, Caruffo M, Paredes R, Ramirez G, et al. Am J Reprod Immunol. 2017 Oct 19. DOI 10.1111/aji.12772. [Epub ahead of print]

Problem Immunocastration or vaccination against the GnRH-I hormone is a promising alternative to reproductive control in different animal species. Given the low immunogenicity of this hormone, the use of adjuvants becomes necessary. Method of study This study evaluated the effects of three adjuvants that induce different immune response profiles over gonadal function, fertility, and expression of GnRH-I. Female mice (n = 6) were vaccinated at days 1 and 30 with a recombinant antigen for immunocastration and different adjuvants that induced preferentially Th1/Th2, Th2, and Th1 immune profiles. Results Th1/Th2 response is the most efficient to block reproductive activity in vaccinated animals, reducing the number of luteal bodies and pre-ovulatory follicles. Th2 and Th1/Th2 responses induced an increase in GnRH-I at the hypothalamus. Conclusions The immune profile induced by different adjuvants is essential on the effects over fertility, gonadal function, and hypothalamic GnRH-I expression in immunocastrated animals.

The influence of different peptide combinations to increase the immunogenicity of the Gonadotrophin Releasing Hormone Vaccine for prostate cancer treatment. Aguilar FF, Barranco JJ, Aguilera LC, Fuentes EB, Serradelo Leal JA, López EH, et al. J Exp Ther Oncol. 2017 Nov;12(2):87–94.

Purpose Therapeutic vaccines, specifically the Gonadotrophin Releasing Hormone (GnRH) vaccine, are considered an additional therapeutic option for advanced stage prostate cancer. Our work showed amplification of the immune response when combining two peptides with and without the Very Small Size Proteoliposomes (VSSP). VSSP is a potent adjuvant for dendritic cells activation and Th1 differentiation as enhanced immune response. Methods The test was carried out in Copenhagen rats as animal model. Results The use of both peptides and their combination with VSSP generated a potentiation of the immune response statistically superior, in term of generating anti GnRH antibody and effects on target organs, when it was compared with the effects which occurs with independent peptides and with and without the VSSP. These results can find application in the development of GnRH vaccine candidates and in peptide based vaccine strategies. Conclusions Immunization with the peptide combination enhances the immune response when mixed with the VSSPs.

Trail Making Test: Normative data for the Latin American Spanish-speaking pediatric population. Arango-Lasprilla JC, Rivera D, Ramos-Usuga D, Vergara-Moragues E, Montero-López E, Adana Díaz LA, et al. NeuroRehabilitation. 2017 Oct 11. DOI: 10.3233/NRE-172247. [Epub ahead of print].

Objective To generate normative data for the Trail Making Test (TMT) in Spanish-speaking pediatric populations. Method The sample consisted of 3,337 healthy children from nine countries in Latin America (Chile, Cuba, Ecuador, Guatemala, Honduras, Mexico, Paraguay, Peru, and Puerto Rico) and Spain. Each participant was administered the TMT as part of a larger neuropsychological battery. The TMT-A and TMT-B scores were normed using multiple linear regressions and standard deviations of residual values. Age, age2, sex, and mean level of parental education (MLPE) were included as predictors in the analyses. Results The final multiple linear regression models showed main effects for age on both scores, such that as children needed less time to complete the test while they become older. TMT-A scores were affected by age2 for all countries except, Cuba, Guatemala, and Puerto. TMT-B scores were affected by age2 for all countries except, Guatemala and Puerto Rico. Models indicated that children whose parents had a MLPE >12 years of education needed less time to complete the test compared to children whose parents had a MLPE ≤12 years for Mexico and Paraguay in TMT-A scores; and Ecuador, Mexico, Paraguay, and Spain for TMT-B scores. Sex affected TMT-A scores for Chile, Cuba, Mexico, and Peru, in that boys needed less time to complete the test than girls. Sex did not affect TMT-B scores. Conclusions This is the largest Spanish-speaking pediatric normative study in the world, and it will allow neuropsychologists from these countries to have a more accurate approach to interpret the TMT in pediatric populations.

Upregulation of HLA Class I Expression on Tumor Cells by the Anti-EGFR Antibody Nimotuzumab. Garrido G, Rabasa A, Garrido C, Chao L, Garrido F, García-Lora ÁM, et al. Front Pharmacol. 2017 Oct 6;8:595. DOI: 10.3389/fphar.2017.00595. eCollection 2017.

Defining how epidermal growth factor receptor (EGFR)-targeting therapies influence the immune response is essential to increase their clinical efficacy. A growing emphasis is being placed on immune regulator genes that govern tumor – T cell interactions. Previous studies showed an increase in HLA class I cell surface expression in tumor cell lines treated with anti-EGFR agents. In particular, earlier studies of the anti-EGFR blocking antibody cetuximab, have suggested that increased tumor expression of HLA class I is associated with positive clinical response. We investigated the effect of another commercially available anti-EGFR antibody nimotuzumab on HLA class I expression in tumor cell lines. We observed, for the first time, that nimotuzumab increases HLA class I expression and its effect is associated with a coordinated increase in mRNA levels of the principal antigen processing and presentation components. Moreover, using 7A7 (a specific surrogate antibody against murine EGFR), we obtained results suggesting the importance of the increased MHC-I expression induced by EGFR-targeted therapies display higher in antitumor immune response. 7A7 therapy induced upregulation of tumor MHC-I expression in vivo and tumors treated with this antibody display higher susceptibility to CD8+ T cells-mediated lysis. Our results represent the first evidence suggesting the importance of the adaptive immunity in nimotuzumab-mediated antitumor activity. More experiments should be conducted in order to elucidate the relevance of this mechanism in cancer patients. This novel immune-related antitumor mechanism mediated by nimotuzumab opens new perspectives for its combination with various immunotherapeutic agents and cancer vaccines.

Vaccine-mismatched influenza B/Yamagata lineage viruses in Cuba, 2012-2013 season. Arencibia A, Piñón A, Acosta B, Fernández L, Muné M, Valdés O, et al. Infect Genet Evol. 2017 Dec 8;58 :110–14.

Annual trivalent influenza vaccines contain one of influenza B lineages; influenza B/Victoria-lineage or influenza B/Yamagata viruses. Theoretically, these vaccines should protect against viruses expected to circulate in the next influenza season. The National Influenza Centers, based on surveillance data from National Reference Laboratories, selects the strains composing each annual trivalent or tetravalent vaccine. Nevertheless, in some epidemics, vaccine strains do not match genetically with circulating strains. The aim of the present study is to compare the HA1-domain of 42 influenza B viruses circulating in Cuba during the 2012-2013 season with the vaccine strain B/Wisconsin/01/2010-like virus from the B/Yamagata lineage, included in the 2012-2013 Northern-Hemisphere Influenza vaccine. The efficacy of the influenza vaccine was also estimated. The analysis of the present study indicates that the B/Victoria and B/Yamagata lineages co-circulated in Cuba in the 2012-2013 season. In 2012-2013 season, according to the sequences analysis, trivalent vaccine did not match with the circulating strains. The present study also detected amino acid substitutions which could have altered the antigenic properties of HA gene. The results presented here suggest the need to consider a possible introduction of tetravalent influenza vaccine in Cuba, as has been recommended by the WHO to ensure higher levels of protection.