Helpful Criteria When Implementing NGS Panels in Childhood Lymphoblastic Leukemia.

The development of Next-Generation Sequencing (NGS) has provided useful diagnostic, prognostic, and therapeutic strategies for individualized management of B-cell precursor acute lymphoblastic leukemia (BCP-ALL) patients. Consequently, NGS is rapidly being established in clinical practice. However, the technology's complexity, bioinformatics analysis, and the different available options difficult a broad consensus between different laboratories in its daily routine introduction. This collaborative study among Spanish centers was aimed to assess the feasibility, pros, and cons of our customized panel and other commercial alternatives of NGS-targeted approaches. The custom panel was tested in three different sequencing centers. We used the same samples to assess other commercial panels (OncomineTM Childhood Cancer Research Assay; Archer®FusionPlex® ALL, and Human Comprehensive Cancer Panel GeneRead Panel v2®). Overall, the panels showed a good performance in different centers and platforms, but each NGS approach presented some issues, as well as pros and cons. Moreover, a previous consensus on the analysis and reporting following international guidelines would be preferable to improve the concordance in results among centers. Our study shows the challenges posed by NGS methodology and the need to consider several aspects of the chosen NGS-targeted approach and reach a consensus before implementing it in daily practice.

Construction of reference chromosome-scale pseudomolecules for potato: integrating the potato genome with genetic and physical maps.

The genome of potato, a major global food crop, was recently sequenced. The work presented here details the integration of the potato reference genome (DM) with a new sequence-tagged site marker-based linkage map and other physical and genetic maps of potato and the closely related species tomato. Primary anchoring of the DM genome assembly was accomplished by the use of a diploid segregating population, which was genotyped with several types of molecular genetic markers to construct a new ~936 cM linkage map comprising 2469 marker loci. In silico anchoring approaches used genetic and physical maps from the diploid potato genotype RH89-039-16 (RH) and tomato. This combined approach has allowed 951 superscaffolds to be ordered into pseudomolecules corresponding to the 12 potato chromosomes. These pseudomolecules represent 674 Mb (~93%) of the 723 Mb genome assembly and 37,482 (~96%) of the 39,031 predicted genes. The superscaffold order and orientation within the pseudomolecules are closely collinear with independently constructed high density linkage maps. Comparisons between marker distribution and physical location reveal regions of greater and lesser recombination, as well as regions exhibiting significant segregation distortion. The work presented here has led to a greatly improved ordering of the potato reference genome superscaffolds into chromosomal "pseudomolecules".

Genome-wide identification and mapping of NBS-encoding resistance genes in Solanum tuberosum group phureja.

The majority of disease resistance (R) genes identified to date in plants encode a nucleotide-binding site (NBS) and leucine-rich repeat (LRR) domain containing protein. Additional domains such as coiled-coil (CC) and TOLL/interleukin-1 receptor (TIR) domains can also be present. In the recently sequenced Solanum tuberosum group phureja genome we used HMM models and manual curation to annotate 435 NBS-encoding R gene homologs and 142 NBS-derived genes that lack the NBS domain. Highly similar homologs for most previously documented Solanaceae R genes were identified. A surprising ∼41% (179) of the 435 NBS-encoding genes are pseudogenes primarily caused by premature stop codons or frameshift mutations. Alignment of 81.80% of the 577 homologs to S. tuberosum group phureja pseudomolecules revealed non-random distribution of the R-genes; 362 of 470 genes were found in high density clusters on 11 chromosomes.

Oscillatory sensory selection mechanisms during intersensory attention to rhythmic auditory and visual inputs: a human electrocorticographic investigation.

Oscillatory entrainment mechanisms are invoked during attentional processing of rhythmically occurring stimuli, whereby their phase alignment regulates the excitability state of neurons coding for anticipated inputs. These mechanisms have been examined in the delta band (1-3 Hz), where entrainment frequency matches the stimulation rate. Here, we investigated entrainment for subdelta rhythmic stimulation, recording from intracranial electrodes over human auditory cortex during an intersensory audiovisual task. Audiovisual stimuli were presented at 0.67 Hz while participants detected targets within one sensory stream and ignored the other. It was found that entrainment operated at twice the stimulation rate (1.33 Hz), and this was reflected by higher amplitude values in the FFT spectrum, cyclic modulation of alpha-amplitude, and phase-amplitude coupling between delta phase and alpha power. In addition, we found that alpha-amplitude was relatively increased in auditory cortex coincident with to-be-ignored auditory stimuli during attention to vision. Thus, the data suggest that entrainment mechanisms operate within a delimited passband such that for subdelta task rhythms, oscillatory harmonics are invoked. The phase of these delta-entrained oscillations modulates alpha-band power. This may in turn increase or decrease responsiveness to relevant and irrelevant stimuli, respectively.

[Low-dose cinacalcet reduces serum calcium in patients with primary hyperparathyroidism not eligible for surgery]. / Dosis bajas de cinacalcet reducen el calcio sérico en pacientes con hiperparatiroidismo primario no subsidiario de tratamiento quirúrgico.

We present our experience with low-dose cinacalcet to normalize serum calcium in patients with primary hyperparathyroidism (PHPT) not eligible for surgery. We analyzed the impact of this drug on various parameters of calcium-phosphorus metabolism and its tolerability profile. We recruited 17 patients diagnosed with PHPT who had hypercalcemia and also met one or more of the following inclusion criteria: elevated risk for parathyroidectomy, persistent/recurrent PHPT after previous parathyroid surgery or refusal to undergo surgery. The starting dose of cinacalcet was 30 or 60 mg/day, which was adjusted depending on the degree of calcemia reduction and tolerance to the drug. We observed a reduction in serum calcium that was already evident in the first post-treatment test. Appropriate dose adjustment was performed when required and normal serum calcium levels were achieved in most patients, remaining stable during follow-up. Parathyroid hormone was reduced but not normalized in most patients. Calciuria decreased while serum phosphate and alkaline phosphatase levels increased. Cinacalcet tolerance was generally good at the doses used. The most common adverse effects were weakness, dizziness and asthenia, leading to treatment withdrawal in only one patient. We conclude that low-dose cinacalcet reduces serum calcium efficiently, normalizes calcium levels in most patients with PHPT not eligible for surgical treatment and has a good tolerability profile.

Day-night differences in membrane-bound pyroglutamyl-2-naphthylamide-hydrolyzing activity in rat hypothalamus, pituitary, and retina.

Membrane-bound pyroglutamyl-2-naphthylamide-hydrolyzing enzyme activity was analyzed fluorometrically in the anterior hypothalamus, pituitary, and retina of adult male rats to investigate day-night differences. Six groups (n=6 per group) were assessed--three during the light span and three during the dark span--under a standard 12 h-12 h light-dark cycle (light on from 07:00 to 19:00 h) and controlled temperature environment, with food and water available ad libitum. In the hypothalamus, enzyme activity levels were higher for time points of the dark than the light period. In contrast, the pituitary and retina exhibited the highest levels at the time points of the light period. The pituitary and retina also exhibited significant differences between the clock-hour means of the light period. Day-night differences in membrane-bound pyroglutamyl-2-naphthylamide-hydrolyzing activity may reflect differences in its susceptible endogenous substrates.

Audio-visual multisensory integration in superior parietal lobule revealed by human intracranial recordings.

Intracranial recordings from three human subjects provide the first direct electrophysiological evidence for audio-visual multisensory processing in the human superior parietal lobule (SPL). Auditory and visual sensory inputs project to the same highly localized region of the parietal cortex with auditory inputs arriving considerably earlier (30 ms) than visual inputs (75 ms). Multisensory integration processes in this region were assessed by comparing the response to simultaneous audio-visual stimulation with the algebraic sum of responses to the constituent auditory and visual unisensory stimulus conditions. Significant integration effects were seen with almost identical morphology across the three subjects, beginning between 120 and 160 ms. These results are discussed in the context of the role of SPL in supramodal spatial attention and sensory-motor transformations.

Oscillatory beta activity predicts response speed during a multisensory audiovisual reaction time task: a high-density electrical mapping study.

Bisensory redundant targets are processed faster than the respective unisensory target stimuli alone as evidenced by substantially faster reaction times (RTs). This multisensory RT facilitation has been interpreted as an expression of integrative processing between the different sensory modalities. However, the neuronal mechanisms underlying the RT facilitation effect are not well understood. Oscillatory responses in the beta frequency range (13-30 Hz) have been related to sensory-motor processing. Here, we investigated whether modulation of beta responses might also underlie the faster RTs seen for multisensory stimuli. Using high-density electrical mapping, we explored the association between early (50-170 ms) multisensory processing in the evoked beta response and RTs recorded during a simple RT task. Subjects were instructed to indicate the appearance of any stimulus in a stream of auditory-alone (A), visual-alone (V), and multisensory (AV) stimuli by a button press. Beta responses were analyzed using Morlet wavelet transformations. Multisensory interactions were found over frontal, occipital, central, and sensory-motor regions. Critically, beta activity correlated with mean RTs over all stimulus types. Significant negative correlations were found for frontal, occipital, and sensory-motor scalp regions. We conclude that the association between oscillatory beta activity and integrative multisensory processing is directly linked to multisensory RT facilitation effects.

Estudio retrospectivo de las reacciones adversas a los medicamentos en dos hospitales de Barquisimeto / Retrospective stdies of the adverse drug reactions in the two hospitals the Barquisimeto

El objetivo de este estudio fue el determinar la frecuencia de Reacciones Adversas Medicamentosas (RAM) en los dos principales hospitales de Barquisimeto, Estado Lara. Los métodos incluyeron una revisión crítica, retrospectiva de registros médicos obtenidos de 62.456 pacientes hospitalizados dados de alta entre 1998 y 2002, del Hospital Pediátrico "Doctor Agustín Zubillaga" (HPDAZ) y el Departamento de Medicina del Hospital Central Universitario "Doctor Antonio María Pineda" (DM-HCUAMP). El análisis fue descriptivo. Se encontraron solamente 37 reportes válidos de RAM en este estudio. Esto implica una frecuencia de reporte de 5,92 RAM por cada 10.000 pacientes egresados, siendo de 7,88 y 5,36 por 10.000 pacientes egresados en el DM-HCUAMP y el HDPAZ respectivamente. La evidencia sugiere un considerable sub-registro de RAM en Barquisimeto y, por tanto es necesario tomar urgentes medidas correctivas de esta situación

Algunos aspectos clínicos-epidemiológicos del cafeinismo en una comunidad rural

Se realizó un estudio descriptivo transversal con el objetivo de determinar la prevalencia y algunos aspectos clínicos-epidemiológicos del cafeinismo en la comunidad rural correspondiente al consultorio 76 del policlínico Ramón Heredia Umpierre de Veguitas, municipio Yara, durante el período 1ro. de julio del 2000 hasta diciembre del 2001. El universo estuvo dado por 62 pacientes identificados como enfermos (cafeinómanos) a partir de la aplicación del mismo modelo diagnóstico utilizado para el alcoholismo; se utilizó como medida de resumen el porcentaje y parámetros como la media y desviación estándar. Existió una alta prevalencia de cafeinismo (12,6 por ciento) y sobre todo fueron más afectadas las mujeres con marcada tendencia a incrementarse en ambos sexos paralelo a la edad. La satisfacción fue el móvil mayormente referido por los pacientes para el consumo de café (41,9 por ciento); se destacan las manifestaciones físicas (principalmente la acidez y al epigastralgia), por encima de las psíquicas, dentro de las cuales la ansiedad figuró como la más importante. En el marco actual de una importante batalla de carácter coyuntural que alcanza su mayor connotación en Cuba, esta investigación puede convertirse en una referencia que da medida de la necesidad de intervenir frente a un hábito tóxico que para los efectos es parte de una vieja costumbre social, ignorando casi siempre su nocividad(AU)

A corneal diffuse neurofibroma as a manifestation of von recklinghausen disease.

PURPOSE: To report a case of a primary corneal diffuse neurofibroma in a patient with von Recklinghausen disease (NF-1). METHODS: Case report. A physical examination and histopathology were performed. The immunohistochemical studies were performed using an avidin-biotin-peroxidase complex technique on formalin-fixed and paraffin-embedded tissue. Histologic sections from corneal tissue were incubated with primary antibodies against vimentin and S-100 protein. A complementary ultrastructural study of the same formalin-fixed and paraffin-embedded tissue was made. RESULTS: The ophthalmologic examination revealed a yellowish-white elevated mass that involved the supratemporal cornea but not the limbus. Histologic study showed a tumor of the peripheral nerve sheath, a diffuse neurofibroma in the corneal stroma, and proliferation of spindle cells with markedly elongated nuclei. Cells comprising the tumor reacted with vimentin and S-100 protein, and the ultrastructural studies revealed myelinated nerve fibers confirming the diagnosis. CONCLUSION: The development of a primary diffuse neurofibroma in the cornea of patients with von Recklinghausen disease is possible. The present case supports the statement that neurofibromas arising from the peripheral nerve sheath may involve any part of the body.

Dietary fatty acid composition affects aminopeptidase activities in the testes of mice.

The autocrine/paracrine control mechanisms of local factors, such as the renin-angiotensin system and the thyrotropin-releasing hormone (TRH), seem to play a relevant role in testicular physiology. It has been proposed that dietary fat composition influences male reproductive function modifying the cholesterol-phospholipid composition of testicular plasma membranes. Modifications in the composition and physical properties of the membranes may lead to alterations in the activities of membrane-bound (M-B) enzymes. We have previously demonstrated that cholesterol and steroid hormones affect aminopeptidase (AP) activities. Dietary fatty acids with different degrees of saturation modified AP activities in the serum of mice and an olive oil supplemented diet influenced the AP activities in the testes of mice. We hypothesized that the modification of dietary fat composition may affect angiotensin- [glutamyl-AP (GluAP), aspartyl-AP (AspAP)] and TRH- [pyroglutamyl-AP (pGluAP)] degrading activities in the testis. In this study, we investigated the effect of diets supplemented with sunflower oil (SFO), fish oil (FO), olive oil (OO), lard (L) or coconut oil (CO) on soluble (Sol) and M-B GluAP, AspAP and pGluAP in mice testis, using arylamides as substrates. Sol GluAP activity did not show differences among groups. However, Sol AspAP and Sol pGluAP progressively decreased with the degree of saturation of the fatty acid used in the diet. In contrast, M-B GluAP progressively increased with the degree of saturation of the fatty acid used in the diet. For M-B AspAP activity, mice fed diets containing FO showed significantly higher levels than those fed diets containing SFO, OO and L but not those containing CO. For M-B pGluAP activity, the highest levels were observed for mice fed diets containing FO and OO. The present data suggest that the type of fat used in the diet may influence the autocrine/paracrine functions of locally synthesized angiotensin peptides and TRH in the testis, and consequently may be important in male reproductive functions.

Ethanol modifies differently aspartyl- and glutamyl-aminopeptidase activities in mouse frontal cortex synaptosomes.

Aminopeptidase A activity (aspartyl aminopeptidase (AspAP) and glutamyl aminopeptidase (GluAP) exerts angiotensinase activity due to its relation to the metabolism of angiotensins in the regional brain renin-angiotensin system (RAS). This activity may also modify the free amino acid pool through the release of N-terminal acidic amino acids. Ethanol (EtOH) exerts profound effects on the brain, inducing important neurological damages. Our purpose is to study the influence of EtOH on AspAP and GluAP activities on basal and K(+)-stimulated conditions, at the synapse level. We used mouse frontal cortex synaptosomes and their incubation supernatant in a Ca(2+)-containing or Ca(2+)-free artificial cerebrospinal fluid. We evaluate the possible contribution of these enzymatic activities on brain blood pressure regulation through RAS and/or the free acidic amino acid pool. The results obtained are correlated with several parameters of oxidative stress, such as free radical generation, lipid peroxidation, and protein oxidation. Under basal conditions, in synaptosomes, EtOH inhibits AspAP and GluAP activities independently of Ca(2+). In the supernatant, however, EtOH differently modulates the two enzyme activities under the various concentrations. Under K(+)-stimulated conditions, EtOH inhibits the K(+)-stimulated increase on AspAP and GluAP differently depending on the presence or absence of Ca(2+) and the concentration of EtOH used. These results invalidate the idea that excess free acidic amino acids could be released by AspAP and GluAP to induce neurodegeneration. The changes in AspAP and GluAP activities as a consequence of EtOH administration and their role in the brain RAS are discussed.

Plantas tóxicas y medicinales cómunes causantes de toxicidad en el Estado Lara, Venezuela / Plant toxic and medicinal commun causated of toxicity in the Lara State, Venezuela

Describimos las plantas que frecuentemente causan toxicidad en la región Centro-Occidental del país. Se identifican con los nombres botáncos, comunes, partes toxicas, principios activos, efectos clínicos y tratamiento. Además se describen plantas con efectos depilatorios y psicotrópicas. El tabaquismo y sus consecuencias.