Evaluación y detección bioinformática de terpenoides con potencial inhibitorio de la proteína viral 1 (VP1) del virus de la bursitis infecciosa / Evaluation and bioinformatic detection of terpenoids with inhibitory potential of viral protein 1 (VP1) of infectious bursitis virus

El virus de la bursitis infecciosa (IBDV) es el agente causal de la enfermedad de la bursa, la cual afecta principalmente a poblaciones avícolas jóvenes y genera un impacto económico negativo en la producción. La proteína viral (VP1) es una enzima con funciones clave para la replicación del genoma viral, por lo que puede ser considerada blanco para la búsqueda de compuestos con posibles actividades inhibitorias. El objetivo de esta investigación fue evaluar terpenoides con potencial inhibitorio de la proteína VP1 del IBDV mediante herramientas de aproximaciones bioinformáticas. Se seleccionó un total de 52 terpenoides, cuyas propiedades farmacológicas, farmacocinéticas y tóxicas (ADME-Tox) se evaluaron. Las moléculas sin actividades tóxicas y con aptitudes farmacocinéticas fueron sometidas a pruebas exhaustivas de acoplamiento molecular con el sitio catalítico de la VP1 mediante el uso del algoritmo genético y de Broyden-Fletcher-Goldfarb-Shanno junto con el método de optimización local de gradientes. Los datos obtenidos revelaron que la Giberelina A1 presenta valores de energía libre de unión significativamente (p< 0,05) favorables (ΔG=-7,28±0,06 kcal/mol; Kdcalc= 8,62±0,99 µM) en comparación con los sustratos rCTP y rGTP. El complejo Giberelina A1-VP1 presenta puentes de hidrógeno con los residuos Arg335 y Asp402, los cuales cumplen roles importantes en la actividad catalítica en la replicación viral. Estos hallazgos sugieren que el terpenoide Giberelina A1 puede ser considerado como compuesto candidato para estudios in vitro de inhibición de funciones de la VP1 e in vivode actividades antivirales contra el virus de la bursitis infecciosa.

Infectious bursitis virus (IBDV) is the infectious agent of bursal disease, which mainly affects young poultry populations, generating a negative economic impact on productions. The viral protein 1 (VP1) is an enzyme with important functions for the replication of the viral genome, so it could be considered as a target for searching compounds with possible inhibitory activities. The aim of this research was to evaluate terpenoids with inhibitory potential of the VP1 protein of IBDV using computational approximations tools. A total of 52 terpenoids were selected and evaluated for their pharmacological, pharmacokinetic and toxic properties (ADME-Tox). Molecules without toxic activities and with pharmacokinetic competences were subjected to extensive molecular docking tests with the catalytic site of VP1 using the genetic and Broyden-Fletcher-Goldfarb-Shanno algorithms with a local gradient optimization method. Data obtained revealed that Gibberellin A1 exhibits significantly (P < 0.05) favorable binding free energy values (ΔG=-7.28±0.06 kcal/mol; Kdcalc= 8.62±0.99 µM) compared to rCTP and rGTP subs-trates. The Gibberellin A1-VP1 complex exhibits hydrogen bonds with residues Arg335 and Asp402, which play important roles in catalytic activity in viral replication. These findings suggest that the terpenoid Gibberellin A1 could be considered as a candidate compound for in vitro studies of inhibition of VP1 functions and in vivo antiviral activities against infectious bursitis virus.

Histoplasmosis diseminada. A propósito de un caso / Disseminated Histoplasmosis. About a case

RESUMEN La histoplasmosis es una micosis profunda causada por el hongo dimorfo Histoplasma capsulatum (H. capsulatum). Ingresa al organismo principalmente por la vía inhalatoria en forma de microconidias, las cuales se transforman en elementos levaduriformes intracelulares, y luego se diseminan por vía hemática. La primoinfección en pacientes inmunocompetentes suele ser asintomática y de resolución espontánea, pero los pacientes inmunodeprimidos generalmente pueden presentar una enfermedad diseminada con compromiso mucocutáneo con pápulas, nódulos, gomas, úlceras de fondo granulomatoso serosanguinolento y costras. Se presenta un caso clínico de un paciente diabético inmunodeprimido con infección por H. capsulatum, en el cual se realiza diagnóstico a partir de las lesiones cutáneas.

SUMMARY Histoplasmosis is a deep mycosis caused by the dimorfo fungus Histoplasma capsulatum (H. capsulatum). Which enters the body mainly through the inhalation route in the form of microconidia which are transformed into intracellular levaduriform elements, and then disseminated by blood. The primary infection in immunocompetent patients is usually asymptomatic and spontaneously resolved, but immunocompromised patients can usually present with a disseminated disease with mucocutaneous involvement, with papules, nodules, gums, granulomatous serosanguinolent fundus ulcers and scabs. A clinical case of an immunocompromised diabetic patient with H. capsulatum infection is presented, in which diagnosis is made from the skin lesions.

Ultrasound versus anatomical landmarks: Immediate complications in the central venous catheterization in children under 18 years of age. / Ultrasonido frente a referencias anatómicas: complicaciones inmediatas en la canalización de vía venosa central en menores de 18 años.

INTRODUCTION: The insertion of a central venous line in children and adolescents is technically more difficult, due to the smaller size of the structures. This can lead to an increase in immediate complications, which can be reduced when using ultrasound. In our institution, the percentage of these complications and the use of ultrasound are unknown. OBJECTIVE: To describe the frequency of immediate complications of central venous catheterisation guided by the ultrasound in a general university hospital, compared to the anatomical landmarks technique in children less than 18years of age. MATERIALS AND METHODS: Observational, retrospective, and analytical study, comparing the frequency of complications with two central venous catheterisation techniques: anatomical landmarks and ultrasound, according to the clinical records of procedures performed from June to November 2016. RESULTS: A total of 201 procedural records were analysed, of which 71% were with landmarks, and 29% with ultrasound. The overall incidence of immediate complications was 18.4%, with 12% using ultrasound and 21% using landmarks (OR: 0.5; 95%CI: 0.2-1.2). Children under 5years of age presented with 90% of the complications, the most frequent being the impossibility of passing the guide (29.7%) and multiple punctures (24.3%). There was no arterial puncture with use of ultrasound. Ultrasound was used by 13.4% of paediatric surgeons, by 32.4% of paediatricians, and 46.4% of anaesthetists, with complications of 25%, 19%, and 7%, respectively. The main indication for catheterisation was the need for vasoactive agents (74%), with the procedure being more complicated in patients with no peripheral venous accesses (46%). The success rate with anatomical landmarks was 77.6%, compared to 91.4% with ultrasound. CONCLUSION: Central venous catheterisation with ultrasound guidance in children under 18 reduces immediate complications by 42.8% and improves the success rate by 13.8%.

Estimating Waiting Time for Deceased Donor Renal Transplantion in the Era of New Kidney Allocation System.

BACKGROUND: On December 4, 2014, a new deceased donor kidney allocation system (KAS) was implemented. The KAS was designed to improve organ equity and graft-recipient longevity matching. However, estimated wait-time to deceased donor transplantation is difficult to predict post-KAS. METHODS: Using the Kidney-Pancreas Simulated Allocation Model software (KPSAM), a program that the Organ Procurement and Transplant Network uses to assess policy proposals, we compared the kidney allocations of both the new (post-KAS) and old policies (pre-KAS) (10 iterations for each group; total N = 204,148) and estimated wait-time based on blood type, duration of dialysis exposure, and calculated panel-reactive antibody (CPRA). RESULTS: The simulations revealed that estimated median (25(th) and 75(th) percentile) waiting time in transplanted recipients decreased from 2.3 (1.2, 3.8) years in the old allocation to 1.8 (0.8, 3.4) years in the new allocation system. The rate of transplantations performed within the first year of wait-listing increased from 20.7% to 31.3%. The KPSAM resulted in more transplantations in recipients with more than 5 years of dialysis exposure (26.5% to 37.4%), longevity matching (12.2% to 17.5%), blood group B (12.6% to 17.2%), and high CPRA ≥98% (1.9% to 4.3%) in post-KAS compared with pre-KAS simulations. CONCLUSIONS: Based on the KPSAM results, it was projected that post-KAS wait-time in transplanted recipients might decrease approximately 6 months (22%) across all CPRA categories. It might be related to the KAS awarding waiting time points for prelisting dialysis time and priority points awarded based on CPRA (bolus effect).

Anomalía de Ebstein de la válvula tricúspide. A propósito de un caso / Ebstein anomaly of the tricuspid valve. Report of a case

Introducción. La anomalía de Ebstein constituye menos del 1% de todas las cardiopatías congénitas. Consiste en el desplazamiento inferior de la válvula tricúspide anómala hacia el ventrículo derecho. Reporte: Paciente varón de 8 años de edad, procedente de Trujillo, con diagnóstico de enfermedad de Ebstein al mes de edad, que ingresa por emergencia presentando cianosis y dolor torácico. Interpretación: En la anomalía de Ebstein la gravedad de los síntomas y el grado de cianosis dependen del grado de desplazamiento de la válvula tricúspide. Los síntomas pueden ser leves o no aparecer hasta la adolescencia o la edad adulta.

Comparison of longer-term outcomes after kidney transplantation between Hispanic and non-Hispanic whites in the United States.

Little is known about the longer-term kidney transplant outcomes in the rapidly growing Hispanic population. Using the United States Renal Data System, we identified 105 250 Caucasian patients who received a first kidney transplant between January 1, 1996 and December 31, 2010. We tested for differences between Hispanic and non-Hispanic patients in the outcomes of (1) mortality, (2) all-cause graft failure, and (3) graft failure excluding death with a functioning graft. We used Cox regression to estimate (with 95% confidence intervals) multivariable-adjusted cause-specific hazard ratios (aHRCS ) for mortality and all-cause graft failure and subdistribution hazard ratios (aHRSD ) accounting for death as a competing risk for graft failure excluding death with a functioning graft. Both mortality [aHRCS = 0.69 (0.65-0.73)] and all-cause graft failure [aHRCS = 0.79 (0.75-0.83)] were lower in Hispanics. The association between Hispanic ethnicity and graft failure excluding death was modified by age (p < 0.003). Compared with non-Hispanic whites, graft failure excluding death with a functioning graft did not differ in Hispanics aged 18-39 years [aHRSD = 0.96 (0.89-1.05)] or aged 40-59 years [aHRSD = 1.08 (1.00-1.16)], but was 13% lower in those aged ≥60 years [aHRSD = 0.87 (0.78-0.98)]. In conclusion, once accounting for differences in overall survival, better graft survival was found in older Hispanic patients, but among not those aged <60 years.

Historia natural del carcinoma ductal invasivo frente a carcinoma lobulillar invasivo. Nuestra experiencia / Natural history of invasive ductal carcinoma versus invasive lobular carcinoma. Our experience

Introducción: La subclasificación histológica del cáncer de mama infiltrante, ya sea ductal (CDI) o lobulillar (CLI) se correlaciona aparentemente con una biología tumoral distinta. Los datos de nuestro análisis m ostraron que los carcinomas lobulillares invasivos son generalmente de tipo luminales, tienen mayor tamaño al momento del diagnóstico clínico y un superior índice de cirugías radicales. Siendo superior la tasa de recurrencia local. Tanto la supervivencia global como la libre de enfermedad no tuvieron diferencia significativa en ambos grupos. Conclusión: La clasificación histológica del cáncer invasivo de mama, ya sea ductal o lobulillar, no se correlaciona con la supervivencia de la enfermedad. Sin embargo, en nuestro trabajo encontramos diferencias en la recidiva local, no así a distancia, lo que podría deberse a un comportamiento biológico diferente.

Proteomic analysis of intestinal mucosa responses to Salmonella enterica serovar typhimurium in naturally infected pig.

Salmonella enterica serovar typhimurium (S. typhimurium) is one of the most frequent Salmonella serotypes isolated from European pigs. Despite the advances in understanding the mechanisms involved in host-pathogen interactions and host cell responses to S. typhimurium, the global change that occurs in naturally exposed populations has been poorly characterized. Here, we present a proteomics study on intestinal mucosa of pigs naturally infected with S. typhimurium, in order to better understand the pathogenesis of salmonellosis and the pathways which might be affected after infection. Samples were analyzed by 2D-DIGE and 44 different proteins exhibited statistically significant differences. The data set was analyzed by employing the Ingenuity Pathway Analysis and the physiological function most significantly perturbed were immunological and infectious disease, cellular assembly and organization and metabolism. The pathways implicated in the porcine immune response to S. typhimurium were gluconeogenesis and Rho GDI/RhoA signaling, and our results suggest that keratins and the intermediate filaments could play an important role in the damage of the mucosa and in the success of infection. The role of these findings in salmonellosis has been discussed, as well as the importance of analyzing naturally infected animals to have a complete picture of the infection. Also, we compared the results found in this work with those obtained in a similar study using experimentally infected animals.

Eficacia de tres tipos de surfactante exógeno en prematuros con enfermedad de membrana hialina / Effectiveness of three types of exogenous surfactant in preterm infants with hyaline membrane disease

Introducción: La enfermedad de membrana hialina es causa importante de mortalidad neonatal. El objetivo de esta investigación fue evaluar la eficacia de tres tipos de surfactante exógeno en prematuros. Pacientes y Método: Estudio de cohorte retrospectiva, en 93 neonatos prematuros, > 24 semanas y > 500 g de peso al nacer, 31 para cada surfactante. La exposición fue la administración de 1ª dosis bovactant (Alveofact®) 50 mg/kg, beractant (Survanta®) 100 mg/kg inicial, y poractant alfa (Curosurf®) 200 mg/kg. Las variables en estudio incluyeron tiempo de ventilación mecánica, tiempo de oxigenoterapia, estancia hospitalaria, necesidad de segunda dosis de surfactante, eventos adversos por la administración del surfactante y complicaciones por prematuridad. Además, se evaluó mortalidad, displasia broncopulmonar (DBP) y mortalidad o DBP. Análisis estadístico mediante Stata® 11.0, empleando X² o Prueba Exacta de Fisher para variables cualitativas y Pruebas ANOVA o Kruskal-Wallis para cuantitativas y riesgo relativo para las asociaciones, todas con su intervalo de confianza de 95%. Resultados: No hubo diferencias para sexo, peso y edad gestacional al nacer entre los 3 grupos. No se hallaron diferencias estadísticamente significativas para tiempo de ventilación mecánica, tiempo de oxigenoterapia, administración de una segunda dosis de surfactante, estancia hospitalaria y complicaciones entre los 3 grupos. Los eventos adversos por administración de surfactante se presentaron para beractant y poractant alfa. Ocurrieron 30 (32,3 por ciento) muertes, 8 (25,8 por ciento) para bovactant, 10 (32,3 por ciento) beractant y 12 (38,7 por ciento) poractant alfa (p > 0,05). La mortalidad y/o DBP ocurrió en 10 (32,2 por ciento) neonatos con bovactant, 10 (32,2 por ciento) con beractant y 14 (45,2 por ciento) con poractant alfa (p > 0,05). Conclusiones: Los resultados primarios y secundarios entre los tres surfactantes evaluados fueron muy similares...

Introduction: Hyaline membrane disease is an important cause of neonatal mortality. The objective of this research is to evaluate the efficacy of three different exogenous surfactants in premature infants. Patients and Method: A retrospective cohort analysis in 93 preterm infants > 24 weeks and birth weight > 500 g was performed, 31 infants for each surfactant. Exposure consisted of the 1st dose of bovactant (Alveofact®) 50 mg/kg, beractant (Survanta®) 100 mg/kg initially, and poractant alfa (Curosurf®) 200 mg/kg. The variables included duration of mechanical ventilation, duration of oxygen therapy, hospital stay, need for second dose of surfactant, adverse events surfactant administration and prematurity complications. Mortality and bronchopulmonary dysplasia (BPD) were evaluated. Statistical analysis was performed using Stata® 11.0, X² or Fisher exact test for qualitative variables and ALNOVA or Kruskal-Wallis tests for quantitative and association relative risk, all with 95 percent confidence level. Results: There were no gender, weight and gestational age differences at birth among the three groups. No statistically significant differences were found regarding duration of mechanical ventilation, duration of oxygen therapy, administration of a second dose of surfactant, hospital stay and complications among the three groups. Adverse events related to surfactant administration occurred for beractant and poractant alpha. There were 30 (32.3 percent) deaths, 8 (25.8 percent) associated to bovactant, 10 (32.3%) to beractant and 12 (38.7 percent) to poractant alpha (p > 0.05). Mortality and/or BDP occurred in 10 (32.2 percent) infants who received bovactant, 10 (32.2 percent) beractant and 14 (45.2 percent) with poractant alpha (p > 0.05). Conclusions: The primary and secondary outcomes among the three surfactants tested were similar, taking into account the limitations of the work.

Evaluation of hippuric acid content in goat milk as a marker of feeding regimen.

Organic producers, traders, and consumers must address 2 issues related to milk: authentication of the production system and nutritional differentiation. The presence of hippuric acid (HA) in goat milk samples has been proposed as a possible marker to differentiate the feeding regimen of goats. The objective of this work is to check the hypothesis that HA could be a marker for the type of feeding regimen of goats by studying the influence of production system (conventional or organic) and feeding regimen (with or without grazing fodder). With this purpose, commercial cow and goat milk samples (n=27) and raw goat milk samples (n=185; collected from different breeds, localizations, and dates) were analyzed. Samples were grouped according to breed, feeding regimen, production system, and origin to compare HA content by ANOVA and honestly significant difference Tukey test at a confidence level of ≥95%. Hippuric acid content was obtained by analyzing milk samples with capillary electrophoresis. This method was validated by analyzing part of the samples with HPLC as a reference technique. Sixty-nine raw goat milk samples (of the total 158 samples analyzed in this work) were quantified by capillary electrophoresis. In these samples, the lowest average content for HA was 7±3 mg/L. This value corresponds to a group of conventional raw milk samples from goats fed with compound feed. The highest value of this group was 28±10 mg/L, corresponding to goats fed compound feed plus grass. Conversely, for organic raw goat milk samples, the highest concentration was 67±14 mg/L, which corresponds to goats fed grass. By contrast, the lowest value of this organic group was 26±10 mg/L, which belongs to goats fed organic compounds. Notice that the highest HA average content was found in samples from grazing animals corresponding to the organic group. This result suggests that HA is a good marker to determine the type of goats feeding regimen; a high content of HA represents a diet based mainly or exclusively on eating green grass (grazing), independently of the production system. Hence, this marker would not be useful for the actual organic policies to distinguish organic milk under the current regulations, because organic dairy ruminants can be fed organic compound feed and conserved fodder without grazing at all.

Regulation of plasma membrane Ca(2+)-ATPase activity by acetylated tubulin: influence of the lipid environment.

We demonstrated previously that acetylated tubulin inhibits plasma membrane Ca(2+)-ATPase (PMCA) activity in plasma membrane vesicles (PMVs) of rat brain through a reversible interaction. Dissociation of the PMCA/tubulin complex leads to restoration of ATPase activity. We now report that, when the enzyme is reconstituted in phosphatidylcholine vesicles containing acidic or neutral lipids, tubulin not only loses its inhibitory effect but is also capable of activating PMCA. This alteration of the PMCA-inhibitory effect of tubulin was dependent on concentrations of both lipids and tubulin. Tubulin (300µg/ml) in combination with acidic lipids at concentrations >10%, increased PMCA activity up to 27-fold. The neutral lipid diacylglycerol (DAG), in combination with 50µg/ml tubulin, increased PMCA activity >12-fold, whereas tubulin alone at high concentration (≥300µg/ml) produced only 80% increase. When DAG was generated in situ by phospholipase C incubation of PMVs pre-treated with exogenous tubulin, the inhibitory effect of tubulin on PMCA activity (ATP hydrolysis, and Ca(2+) transport within vesicles) was reversed. These findings indicate that PMCA is activated independently of surrounding lipid composition at low tubulin concentrations (<50µg/ml), whereas PMCA is activated mainly by reconstitution in acidic lipids at high tubulin concentrations. Regulation of PMCA activity by tubulin is thus dependent on both membrane lipid composition and tubulin concentration.

Plasma membrane and vesicular glutamate transporter expression in chromaffin cells of bovine adrenal medulla.

The study of the functional expression of glutamate signaling molecules in peripheral tissues has received relatively little attention. However, evidence is increasing for a role of glutamate as an extracellular signal mediator in endocrine systems, in addition to having an excitatory amino acid neurotransmitter role in the CNS. Chromaffin cells are good models of catecholaminergic neurons, in which previous work from our group demonstrated the existence of both functional glutamate receptors and specific exocytotic and nonexocytotic glutamate release. In this work, the presence of specific plasma membrane (EAATs) and vesicular glutamate (VGLUTs) transporters has been investigated by using confocal microscopy, flow cytometric analysis, Western blot, and qRT-PCR techniques. We found specific expression of EAAT3, EAAT2, VGLUT1, and VGLUT3 in about 95%, 65%, 55%, and 25%, respectively, of the whole chromaffin cell population. However, chromaffin cells do not express VGLUT2 and have a very low expression of EAAT1. VGLUTs are localized mainly in the membrane fraction, and EAATs share their subcellular location between membrane and cytosolic fractions. Their estimated molecular weights were about 70 kDa for EAAT2, about 65 kDa for EAAT3, about 50 kDa for VGLUT1, and about 60 kDa for VGLUT3. RT-qPCR techniques confirm the expression of these glutamate transporters at the mRNA level and show a different regulation by cytokines and glucocorticoids between VGLUT1 and -3 and EAAT2 and -3 subfamilies. These interesting results support the participation of these glutamate transporters in the process of glutamate release in chromaffin cells and in the regulation of their neurosecretory function in adrenal medulla.

Involvement of NMDA receptor in the modulation of excitatory and inhibitory amino acid neurotransmitters release in cortical neurons.

The purpose of this paper was to examine the function of N-methyl-D-aspartate (NMDA) glutamate receptor in cortical neurons on amino acid neurotransmitters release as well as the fraction of neurons implicated in the response of this receptor. Local stimulation of these cells at different concentrations of NMDA, agonist of this ionotropic glutamate receptor, produced a dose dependent release of aspartate, glutamate, glycine and GABA. These effects were blocked by DAP5, an antagonist of the NMDA receptor. The amino acid Ca(2+) dependent release mediated by the NMDA receptor, is induced by the opening of voltage-dependent Ca(2+) channels that this receptor promotes. Ca(++) movements were explored in single cells loaded with fura-2. When single cells were stimulated with 100 microM NMDA, the calcium recording performed showed that 82% of the cells responded to this agonist increasing the intracellular calcium concentration, although the amplitude of these increments was variable. The results suggest that NMDA-elicited neurotransmitter release from cortical neurons involves Ca(2+)-dependent and Ca(2+)-independent components, as well as neuron depolarisation, and different VDCC subtypes of N, P/Q or L depending of the amino acid neurotransmitter release elicited by this receptor.

Innate immune activation of swine intestinal epithelial cell lines (IPEC-J2 and IPI-2I) in response to LPS from Salmonella typhimurium.

The innate immune system has the basic function of identifying and eradicating microbial invaders and alerting the adaptative immune system to their presence. In this study, the porcine intestinal innate immune response was evaluated by analysing the expression of TLRs, cytokines and chemokines in two porcine epithelial cell lines from different regions: IPEC-J2 (jejunum) and IPI-2I (ileum). Both cells lines were stimulated with 1microg of LPS from Salmonella typhimurium. RNA was collected at 30min, 1, 2, 3 and 4h after treatment. Expression of TLR-1, -2, -3, -4, -6, -8, -9, -10, TNF-alpha, IL-1beta, -8 and MCP-1 was quantified relative to the quantity of Cyclophilin-A mRNA using real-time quantitative PCR (RTQ-PCR). The results obtained show up differences in the gene expression between both cell lines IPEC-J2 and IPI-2I as response to LPS from S. typhimurium during the activation time, which may suggest an in vivo variability in the innate immune response against pathogens in different regions of the host's gut.

Development of a minimally invasive epidermal abrasion device for clinical skin sampling and its applications in molecular biology.

A new abrasion tool (US patent US7087063 B2) has been developed for collecting skin epidermal samples. This device includes a central shaft that holds the probe in a split chuck. Of the variety of probe designs tested, the laser-cut hollow tube (HT) probe abraded the basal layer of the epidermis most consistently, resulting in representative epidermal skin samples. Compared with traditional clinical methods, the abrasion method allows for high-throughput epidermal skin collection with minimal invasiveness to the volunteer subjects. A large number of abrasion samples have been collected in various clinical studies with no adverse effects observed. Epidermal abrasion, when used appropriately and with the optimized probes, can yield high quality tissue samples that are representative of the epidermis. A sufficient quantity of RNA and protein can be obtained for many subsequent molecular and biochemical applications. Because of its minimal invasiveness and high-throughput nature, the abrasion method can be a valuable tool used to investigate the efficacy of topical applications of skin care products.

A phase II study of epigenetic therapy with hydralazine and magnesium valproate to overcome chemotherapy resistance in refractory solid tumors.

BACKGROUND: Epigenetic aberrations lead to chemotherapy resistance; hence, their reversal by inhibitors of DNA methylation and histone deacetylases may overcome it. PATIENTS AND METHODS: Phase II, single-arm study of hydralazine and magnesium valproate added to the same schedule of chemotherapy on which patients were progressing. Schedules comprised cisplatin, carboplatin, paclitaxel, vinorelbine, gemcitabine, pemetrexed, topotecan, doxorubicin, cyclophosphamide, and anastrozole. Patients received hydralazine at 182 mg for rapid, or 83 mg for slow, acetylators, and magnesium valproate at 40 mg/kg, beginning a week before chemotherapy. Response, toxicity, DNA methylation, histone deacetylase activity, plasma valproic acid, and hydralazine levels were evaluated. RESULTS: Seventeen patients were evaluable for toxicity and 15 for response. Primary sites included cervix (3), breast (3), lung (1), testis (1), and ovarian (7) carcinomas. A clinical benefit was observed in 12 (80%) patients: four PR, and eight SD. The most significant toxicity was hematologic. Reduction in global DNA methylation, histone deacetylase activity, and promoter demethylation were observed. CONCLUSIONS: The clinical benefit noted with the epigenetic agents hydralazine and valproate in this selected patient population progressing to chemotherapy' and re-challenged with the same chemotherapy schedule after initiating hydralazine and valproate' lends support to the epigenetic-driven tumor-cell chemoresistance hypothesis (ClinicalTrials.gov Identifier: NCT00404508).

Self-assembled monolayer-based piezoelectric flow immunosensor for the determination of canine immunoglobulin.

A simple, highly sensitive immunosensor for the direct determination of immunoglobulin (Ig) in canine serum based on a piezoelectric crystal accommodated in a flow-cell was developed and optimized. The new biosensor is also useful for discriminating between Ig subclasses present in canine serum by using specific monoclonal antibodies binding to the coated crystal. Various canine monoclonal anti-IgG were deposited onto the surface of the gold-coated crystal resonator using the self-assembly technique to form a receptor layer. The highly ordered self-assembled monolayers thus obtained provide a well-controlled surface structure and many advantages as regards sensing performance. The results obtained with the proposed immunosensor were compared with those provided by a protein A-based orientation-controlled immobilization method for the same monoclonal antibodies and also with direct physical adsorption of the antibodies. The crystal was accommodated in a flow-cell that was inserted into a buffer flowing stream in order to make resonant frequency measurements.

Within-subject reliability and inter-session stability of EEG power and coherent activity in women evaluated monthly over nine months.

OBJECTIVE: Quantitative EEG parameters during resting conditions are used as baseline in research on cognition and in serial-EEG recordings. Despite its increasing use in cognitive research and the numerous evidences of the existence of sex differences in EEG, EEG stability has been mainly investigated in men. Particularly, studies on stability of coherent activity are scarce. The aim of this study was to investigate within-subject reliability and inter-session stability of resting EEG over a nine-month period in women. METHODS: Within-subject reliability and inter-session stability were analyzed for absolute power and inter- and intrahemispheric coherent activity at central and posterior regions, once a month, in resting conditions, with eyes open and closed. RESULTS: Within-subject reliability was very high (r>0.89) for all subjects and EEG parameters. Inter-session stability was higher with eyes closed and for interhemispheric coherent activity, and poorer with eyes open especially in the alpha band. CONCLUSIONS: Present results indicate high reliability of the pattern of power and coherent activity of each individual woman during rest, and group stability of EEG activity with eyes closed at least over a nine-month period. SIGNIFICANCE: These results provide information on EEG stability in women over a long period.