New microsatellite markers for multi-scale genetic studies on Phlebotomus ariasi Tonnoir,vector of Leishmania infantum in the Mediterranean area

The population structure of Phlebotomus ariasi, a proven vector of Leishmania infantum in the Mediterranean area, is still poorly understood. Previously, only two microsatellite loci had been developed to study the population genetics of this species. Herein we use these loci and determined fourteen novel microsatellite loci, useful for the characterization of P. ariasi populations. These loci were tested on three populations of P. ariasi, two from France and one from Portugal. In addition, the usefulness of these markers was also evaluated on seven other sandfly species.     

Viabilidad de las muestras ganglionares obtenidas por ecobroncoscopia para el estudio de alteraciones epigenéticas en pacientes con cáncer de pulmón

Introduction

The diagnosis of microscopic lymph node metastasis in lung cancer is challenging despite the constant advances in tumor staging. The analysis of the methylation status of certain genes in lymph node samples could improve the diagnostic capability of conventional cyto-histological methods. The aim of this study was to demonstrate the feasibility of methylation studies using cytological lymph node samples.

Methods

Prospective study including 88 patients with a diagnosis or strong suspicion of non-small cell lung cancer, in which an echobronchoscopy was performed on mediastinal or hilar lymph nodes for diagnostic and/or staging. DNA was extracted from cytological lymph node samples and sodium bisulfite modification was performed. Methylation studies for p16/INK4a and SHOX2 were accomplished by MS-qPCR and pyrosequencing.

Results

The methodology used in our study yielded optimal/good DNA quality in 90% of the cases. No differences in DNA concentration were observed with respect to the lymph node biopsied and final diagnosis. Methylation analyses using MS-qPCR and pyrosequencing were not possible in a small number of samples mainly due to low DNA concentration, inadequate purity, fragmentation and/or degradation as a consequence of bisulfite conversion.

Conclusion

Methylation quantification using MS-qPCR and pyrosequencing of cytological lymph node samples obtained using echobronchoscopy is feasible if an appropriate DNA concentration is obtained, notably contributing to the identification of epigenetic biomarkers capable of improving decision-making for the benefit of potentially curable lung cancer patients.     

Perceptions of nursing: a study involving nurses,nursing students,patients and non-nursing students

This paper describes a study of perceptions of nursing by nursing students, qualified nurses (United Kingdom and Spain), patients and non-nursing students. A survey method was used with a longitudinal panel element incorporating a 35-item version of the Nursing Dimensions Inventory translated into Spanish. Data were analysed using principal component analysis, congruence analysis, Mokken scaling, correlation, ANOVA, t-test and Cronbach's alpha. Patients may perceive nursing differently from nursing students, nurses (in the UK and Spain) and non-nurses. These findings are discussed in the context of nurse education and patient care.     

Arcaine and MK-801 make recall state-dependent in rats

RATIONALE: Conditioned fear to context causes freezing and cardiovascular changes in rodents and has been used to measure anxiety. It also activates the dorsolateral column of the periaqueductal gray (dlPAG). Microinjections of cannabinoid agonists into the dlPAG produced anxiolytic-like effects in the elevated plus maze, but the effects of these treatments on fear conditioning remains unknown. OBJECTIVE: The objective of this study was to verify if intra-dlPAG injection of the CB1 cannabinoid receptor agonist anandamide (AEA) or the anandamide transport inhibitor AM404 would attenuate behavioral (freezing) and cardiovascular (increase of arterial pressure and heart rate) responses of rats submitted to a contextual fear-conditioning paradigm. MATERIALS AND METHODS: Male Wistar rats with cannulae aimed at the dlPAG were re-exposed to a chamber where they had received footshocks 48 h before. Fifteen minutes before the test, the animals received a first intra-dlPAG injection of vehicle or AM251, a CB1 receptor antagonist (100 pmol/200 nl), followed 5 min later by vehicle, AEA (5 pmol/200 nl) or AM404 (50 pmol/200 nl). Freezing and cardiovascular responses were recorded for 10 min. RESULTS: Freezing and cardiovascular responses were reduced by administration of either AEA or AM404 into the dlPAG before re-exposition to the aversively conditioned context. These effects were abolished when the animals were locally pretreated with AM251. The latter drug, even at a higher dose (300 pmol), was ineffective when administered alone into the dlPAG. CONCLUSION: The results suggest that facilitation of endocannabinoid-mediated neurotransmission in the dlPAG, through activation of local CB1 receptors, attenuates the expression of contextual fear responses.     

Effect of the Structural and Morphological Properties of Surfactant-Assisted Hydroxyapatite on Dermal Irritation and Antibacterial Activity

Hydroxyapatite (HAp) nanoparticles with a homogeneous rod morphology were successfully synthesized using the hydrothermal method. The powders were characterized using Fourier transform infrared spectroscopy, X-ray diffraction, and scanning electron microscopy. The antibacterial and dermal irritation analyses of the samples were performed and discussed. The use of cationic and anionic surfactants, namely, cetyltrimethylammonium bromide (CTAB) and sodium dodecyl sulfate (SDS), respectively, at a low concentration (2.5 mol%) modified the length/diameter (L/D) ratio of the HAp rods. Structural characterizations of hydroxyapatite synthesized without surfactant (HA), with 2.5 and 5 mol% of SDS (SDS− and SDS+, respectively), and with 2.5 and 5 mol% of CTAB (CTAB− and CTAB+, respectively) revealed well-crystallized samples in the hexagonal phase. The CTAB− sample presented antibacterial activity against Pseudomonas aeruginosa, Escherichia coli, Streptococcus anginosus, Staphylococcus aureus, Micrococcus luteus, and Klebsiella pneumoniae, suggesting that antimicrobial susceptibility was promoted by the bacterial nature and the use of the surfactant. Dermal irritation showed no clinical signs of disease in rabbits during the study, where there was neither erythema nor necrosis at the inoculation sites.     

Del(1)(q23) in a patient with Hutchinson‐Gilford progeria

A 9‐year‐old patient with the classical clinical picture of Hutchinson‐Gilford progeria (HGP) is described. The karyotype shows a 46,XY,del(1)(q23) constitution. Our findings suggest that the interval 1q23 may play a roll in the etiology of HGP. A perturbation in glycosylation in connective tissue has been demonstrated in patients with this condition. This abnormality may be due to a defect in the UDP‐galactose:β‐N‐acetylglucosamina‐β‐1,4‐galactosyltransferase 3 (B4GALT3) gene that has been mapped in the interval 1q21‐23. The cytogenetical analyses of this patient suggest that the B4GALT3 gene could be involved in the pathogenesis of HGP. © 2002 Wiley‐Liss, Inc.     

Genotypic and Phenotypic Analyses of Hepatitis C Virus from Patients Treated with JTK-853 in a Three-Day Monotherapy

JTK-853, a palm site-binding NS5B nonnucleoside polymerase inhibitor, shows antiviral activity in vitro and in hepatitis C virus (HCV)-infected patients. Here, we report the results of genotypic and phenotypic analyses of resistant variants in 24 HCV genotype 1-infected patients who received JTK-853 (800, 1,200, or 1,600 mg twice daily or 1,200 mg three times daily) in a 3-day monotherapy. Viral resistance in NS5B was investigated using HCV RNA isolated from serum specimens from the patients. At the end of treatment (EOT) with JTK-853, the amino acid substitutions M414T (methionine [M] in position 414 at baseline was replaced with threonine [T] at EOT), C445R (cysteine [C] in position 445 at baseline was replaced with arginine [R] at EOT), Y448C/H (tyrosine [Y] in position 448 at baseline was replaced with cysteine [C] or histidine [H] at EOT), and L466F (leucine [L] in position 466 at baseline was replaced with phenylalanine [F] at EOT), which are known to be typical resistant variants of nonnucleoside polymerase inhibitors, were observed in a clonal sequencing analysis. These substitutions were also selected by a treatment with JTK-853 in vitro, and the 50% effective concentration of JTK-853 in the M414T-, C445F-, Y448H-, and L466V-harboring replicons attenuated the susceptibility by 44-, 5-, 6-, and 21-fold, respectively, compared with that in the wild-type replicon (Con1). These findings suggest that amino acid substitutions of M414T, C445R, Y448C/H, and L466F are thought to be viral resistance mutations in HCV-infected patients receiving JTK-853 in a 3-day monotherapy.