[Immunohistochemistry using clone VE1 is an economic, specific and sensitive method for detecting the presence of BRAFV600E mutations in melanoma]. / L'immunohistochimie (clone VE1) est une méthode économique, spécifique et sensible pour détecter la présence d'une mutation BRAFV600E dans le mélanome.

BACKGROUND: Determination of BRAF mutation status is mandatory in the management of patients with inoperable stage IIIC or stage IV melanoma. Currently, molecular biology (MB) has been validated for detecting the presence of BRAF mutations. OBJECTIVE: To compare the sensitivity, specificity and cost of immunohistochemistry (IHC) (clone VE1) versus BM methods (qPCR and Sanger sequencing). PATIENTS AND METHODS: All the samples for which BRAF mutation status was requested between March 2013 and February 2015 at the cellular and molecular analysis laboratory of the Angers Hospital were included retrospectively and consecutively. The IHC (clone VE1) and BM analyses were performed with the same formalin-fixed paraffin embedded tumour samples. The cost of these two methods was determined on the basis of the cost for the French Health Insurance. RESULTS: Two hundred and seven samples were subjected to a determination of BRAF mutational status in IHC and BM. Only one sample was discordant between these two methods (positive in IHC, negative in BM). The sensitivity and specificity of the IHC was 100% and 99.25% respectively. The ratio of the cost of IHC/BM testing was 1:2.1. CONCLUSION: IHC (clone VE1) is a specific, sensitive and economic method for determining BRAFV600E mutation status. Nevertheless, this method must be validated in order to be integrated into a decisional algorithm, alongside BM methods, to determine whether targeted BRAF-inhibitor therapy is indicated.

Perspectives of drug-based neuroprotection targeting mitochondria.

Mitochondrial dysfunction has been reported in most neurodegenerative diseases. These anomalies include bioenergetic defect, respiratory chain-induced oxidative stress, defects of mitochondrial dynamics, increase sensitivity to apoptosis, and accumulation of damaged mitochondria with instable mitochondrial DNA. Significant progress has been made in our understanding of the pathophysiology of inherited mitochondrial disorders but most have no effective therapies. The development of new metabolic treatments will be useful not only for rare mitochondrial disorders but also for the wide spectrum of common age-related neurodegenerative diseases shown to be associated with mitochondrial dysfunction. A better understanding of the mitochondrial regulating pathways raised several promising perspectives of neuroprotection. This review focuses on the pharmacological approaches to modulate mitochondrial biogenesis, the removal of damaged mitochondria through mitophagy, scavenging free radicals and also dietary measures such as ketogenic diet.

[Pituitary adenomas invading the cavernous sinus. Transcranial transcavernous approach]. / Adenomas hipofisarios con invasiòn intracavernosa. Resultados del abordaje trancraneal al seno cavernoso.

UNLABELLED: Pituitary adenomas represent a significant proportion (+o-13%) of all intracranial tumors. Surgical treatment is as rule performed by transsphenoidal approach. However, a small but not insignificant subgroup of pituitary adenomas (5 to 7%) invade the lateral parasellar structures and the cavernous sinus in particular, and poses obvious problems to the therapeutic strategy, since transsphenoidal removal of these adenomas is usually incomplete. OBJECTIVE: To evaluate the results of transcranial removal of a consecutive series of pituitary adenomas invading the cavernous sinus. MATERIAL AND METHOD: A retrospective study of 9 patients harbouring pituitary adenomas invading the cavernous sinus who had been submitted to transcranial surgery was conducted. These patients were operated on in the time span June 1999 - December 2003, in the Nuova Clinica Latina (now NCL- Neurological Centere of Latium), Rome Italy, and the Hospital "Hermanos Ameijeiras", La Habana, Cuba, using a fronto-orbitozigomatic (FOZ) craniotomy, anterior clinoidectomy and a limited dissection of the cavernous sinus through the antero-medial triangle. RESULTS: Total macroscopical removal was achieved in 8 cases, partial in 1, who showed improvement or normalization of the hormonal levels and of the preoperative symptoms. Transitory complications occurred in 6 patients, one of them showed a permanet neurological deficit (paresis of the III cranail nerve). CONCLUSIONS: Transcranial transcavernous approach is an effective technique for attempting total removal of intracavernous pituitary adenomas. It allows to achieve remission of symptoms and hormonal control in the medium-long term. However complications are relatively frequent, and permanent sequelae are not negligible.

Subunit interactions and immobilised dimers of human liver arginase.

Incubation of soluble human liver arginase (L-arginine amidinohydrolase, EC 3.5.3.1) with p-hydroxymercuribenzoate resulted in the dissociation of the enzyme into active dimers. Addition of 2-mercaptoethanol resulted in the regeneration of the tetrameric enzyme. When arginase, bound covalently to nylon, was incubated with p-hydroxymercuribenzoate, matrix-bound dimers were obtained. Incubation of these species with 2-mercaptoethanol resulted in stable, unmodified dimers. Based on this dissociation of arginase, a model with D2-symmetry is suggested for this enzyme. The specific activity, the Km value for arginine, pH optimum and the inhibition constants for ornithine and lysine were determined for monomeric, dimeric and tetrameric forms. It is concluded that the behaviour of the active sites of the monomers is not substantially altered by the interaction of these species in the oligomeric molecule.

[Deep venous thrombosis: an early manifestation of Behçet's disease in childhood]. / Trombosis venosa profunda como manifestación precoz de la enfermedad de Behçet en la infancia.

Behcet's disease is a multisystemic vasculitis of unknown origin whose classical triad is oral and genital ulcers and uveitis. Deep venous thrombosis may sometimes be the first manifestation of this disease but is very rare in children. We report the case of a 14-year-old boy with deep venous thrombosis of the leg and a history of severe aphthous gingivostomatitis, perianal ulcers, Henoch-Schonlein purpura, ecchymotic lesions on the lower limbs and intermittent swelling of the left ankle. Complementary tests ruled out the main causes of thrombosis in children. HLA-B5 and B-51 study proved positive, which, together with the patient's history, suggested a diagnosis of Behcet's disease. This disease usually presents around the third decade of life and is very unusual in children. This case is exceptional because thrombosis as an early manifestation of Behcet's disease is very rare in children and because it led to the diagnosis of Behcet's disease in our patient.

[Delays in the arrival of patients with subarachnoid haemorrhage at a tertiary hospital]. / Demora en la llegada de pacientes con hemorragia subaracnoidea a un hospital terciario.

INTRODUCTION: Patients with subarachnoid haemorrhage should be referred to a tertiary hospital as early as possible. The aim of this research was to study how long these patients take to reach a tertiary care centre in Havana. PATIENTS AND METHODS: We conducted a prospective study of patients with subarachnoid haemorrhage who were admitted to the Stroke Unit at the Hospital Hermanos Ameijeiras between January 2005 and December 2007. The time of arrival at the hospital was determined and a comparison was carried out between patients who arrived after 72 hours and those who reached the hospital sooner. RESULTS: The sample studied comprised 94 patients. Only 34 (36%) arrived during the first three days, whereas 13 (13.8%) reached the hospital within the first 24 hours. None of the sociodemographic and clinical variables that were studied was associated with early remission. The length of time spent in hospital by patients admitted during the first 72 hours was 14.9 days, while in the other cases it was 17.57 days (p = 0.248). The greatest impact on early remission to the tertiary centre was on the outcome at discharge on the Rankin scale, which was lower in those who arrived early compared to the rest of the cases (p = 0.05); the same was true of mortality, which was 5.9% in those who arrived within the first 72 hours versus 11.7% in the others (p = 0.04). CONCLUSIONS: Only a third of the cases reached hospital during the first 72 hours and this group had a more favourable course.

Intracisternal A particles: RNA expression and DNA methylation in murine teratocarcinoma cell lines.

Two main intracisternal-A-particle-specific RNA species (29 to 30S and 35 to 36S) are variably expressed in particle-producing and particle-nonproducing murine teratocarcinoma cell lines. An analysis of DNA methylation patterns after hybridization with an intracisternal-A-particle-specific probe showed an apparent direct correlation between DNA methylation and RNA expression. However, when the methylation assay was performed on the excised intracisternal A particle genes in one of the particle-rich cell lines (PCC6), some undermethylated sequences were detectable. These results are consistent with the concept that only few of the genes are transcriptionally active.

[Formulation, fabrication, and evaluation of a cow's milk formula to Complementary Feeding National Program in Chile]. / Formulación, fabricación y evaluación de una fórmula láctea para el Programa Nacional de Alimentación Complementaria en Chile.

The complementary feeding national programme (PNAC) delivers 2 kg-month of dried full-fat cow's milk (LP) to 0-1 year old infants. The purpose of this work was to design, produce and evaluate nutrient stability and shelf life of a modified LP milk formula (LPM), which seems nutritionally more adequate for infants and is of similar cost than LP, to be included into the PNAC. LPM has the following attributes: P% 12.0 (LP 21.8%): casein/whey proteins ratio: 40 to 50/60 to 50 (LP 80/20); more digestible fats; essential fatty acids: > or = 4% of total calories; vitamins and minerals which cover 100% daily requirements for 0 to 6 months old infants; protein and energy can be modulated between 1.5 and 2.25 g/100 ml and 50-75 kcal/100 ml respectively. Forty tons of LPM were produced in accordance with our specifications for a field study. Quality controls on the product showed the production feasibility of such a formula at a lower price than LP. To determine shelf life, periodical analysis of peroxides, available lysine, ascorbic acid and linoleic acid were carried out in LPM stored for 1 year at 20 degrees C and 37 degrees C. It was found that LPM has an optimum shelf life of 7 months when packed in the same material that is now being in use for LP.(ABSTRACT TRUNCATED AT 250 WORDS)

Study of the evolution of masked bifascicular block.

Masked bifascicular block (MBB) (absent or minimal S in I and VL, high R in Vi and AQRS approximately -60 degrees) is a rare condition. We have found 16 such cases in the last 12 years, thirteen male and three female, mean age 70 +/- 9 years. The conventional electrocardiogram was tabulated and the clinical features and evolution of the patients were followed over 39.1 +/- 32 months. Ten patients required pacemakers, nine because of complete atrioventricular (AV) block or increase in the previous AV block. Seven patients died (follow-up 27.3 +/- 32 months), five from heart failure. This study shows: (1) MBB is a subgroup of patients with a high risk of advanced atrioventricular block; (2) a pacemaker implant does not significantly reduce the high mortality in this group because of the severity of the underlying disease; (3) these results should be evaluated in a larger series of patients in order to confirm our provisional results.

Regulation of intracisternal A particles in mouse teratocarcinoma cells: involvement of DNA methylation in transcriptional control.

The methylation state of Intracisternal A Particle (IAP) genes in mouse teratocarcinoma cell lines has been investigated as an approach to study the regulation of the expression of these particles. Treatment of the cells with the methylation inhibitor 5-azacytidine induces the production of the particles in all the cells; the induction is particularly striking in an embryonal carcinoma cell line which is normally devoid of IAPs. The induction is accompanied by decrease in DNA methylation as demonstrated by using the methylation sensitive isoschizomer enzymes MspI and HpaII. Hypomethylation of the IAP genes correlates with accumulation of IAP, specific polyadenylated RNA reinforcing the hypothesis that methylation plays an important role in the control of IAP expression.

Holter ECG study of the electrocardiographic phenomena in Prinzmetal angina attacks with emphasis on the study of ventricular arrhythmias.

The ECG phenomena in 20 outpatients (121 episodes) suffering from variant angina with transient ST segment elevation greater than 1.5 mm. (Prinzmetal angina) were studied by Holter monitoring. The most important changes in the ECG morphology were: a) increased height of the R wave in all cases, b) the S wave decreased or disappeared, c) the ST segment elevation varied from 1.5 to 38 mm, d) the TQ interval was ascending in 78 episodes, e) there was a double alternance of ST-TQ in 20 episodes and f) the first modification of the ECG was an increase of the T wave height. Arrhythmias were seen in 19 patients (44 episodes). The most frequent were premature ventricular contractions. The prevalence and importance of the ventricular arrhythmias were statistically related to the duration of the episodes (p less than 0.005), the degree of the ST segment elevation (p less than 0.005), the presence of ST-TQ alternance (p less than 0.005) and the presence of increased R wave greater than 25% (p less than 0.025).

Activation of intracisternal a particles by 5-azacytidine in mouse Ki-BALB cell line.

5-Azacytidine activates the production of intracisternal A particles (IAPs) in mouse Ki-BALB cell line as ascertained by electron microscopy scanning and numeration. Efficiency of the activation is higher than that obtained with iododeoxyuridine. The increase in particle production is concentration and time dependent. The results obtained in our system correlate the high IAP expression after drug treatment with a demethylation of IAP-related genes sequences.

Biochemical characterization of endogenous type C virus information in differentiated and undifferentiated murine teratocarcinoma-derived cell lines.

Undifferentiated teratocarcinoma cells express sixfold-higher levels of endogenous xenotropic type C virus-related RNA than differentiated cells. Three species of polyadenylated viral RNA (35S, 24S, and 14S) have been identified in the undifferentiated teratocarcinoma cells. Paradoxically, neither viral particles nor viral proteins have been detected in these cells.