What we see is what we touch? Sex estimation on the skull in virtual anthropology.

BACKGROUND: The increased use of virtual bone images in forensic anthropology requires a comprehensive study on the observational errors between dry bones and CT reconstructions. Here, we focus on the consistency of nonmetric sex estimation traits on the human skull. MATERIALS AND METHODS: We scored nine nonmetric traits on dry crania and mandibles (n = 223) of archaeological origin and their CT reconstructions. Additionally, we 3D surface scanned a subsample (n = 50) and repeated our observations. Due to the intricate anatomy of the mental eminence, we split it into two separate traits: the bilateral mental tubercles and the midsagittal mental protuberance. We provide illustrations and descriptions for both these traits. RESULTS: We obtained supreme consistency values between the CT and 3D surface modalities. The most consistent cranial traits were the glabella and the supraorbital margin, followed by the nuchal crest, zygomatic extension, mental tubercles, mental protuberance, mental eminence, mastoid process and ramus flexure, in descending order. The mental tubercles show higher consistency scores than the mental eminence and the mental protuberance. DISCUSSION: The increased interchangeability of the virtual modalities with each other as compared to the dry bone modality could be due to the lack of tactility on both the CT and surface scans. Moreover, tactility appears less essential with experience than a precise trait description. Future studies could revolve around the most consistent cranial traits, combining them with pelvic traits from a previous study, to test for accuracy.

Periodontal pockets: Predictors for site-related worsening after non-surgical therapy-A long-term retrospective cohort study.

AIM: To evaluate site-related changes in periodontal pocket depth (PPD) after non-surgical periodontal therapy and to identify predictors for PPD changes in a retrospective patient data analysis. MATERIALS AND METHODS: PPD, clinical attachment level, bleeding on probing, tooth mobility (TM), furcation involvement (FI), abutment status, adherence to supportive periodontal care (SPC) and SPC follow-ups were obtained from fully documented patient data before periodontal therapy (baseline, T0), after active periodontal therapy (APT, T1) and during SPC (T2). PPD changes were classified into deteriorated or unchanged/improved at the site level. Multi-level logistic regression analysis was performed to identify factors influencing PPD changes during SPC. RESULTS: This retrospective study included 51 females and 65 males (mean T0 age: 54.8 ± 10.1 years, 25 smokers, 12 diabetics) suffering from Stage III/IV periodontitis. Evaluation outcome: T0/16,044 sampling sites/2674 teeth; T1/15,636/2606; T2/14,754/2459. During 9.0 ± 2.3 years SPC, PPD decreased (-1.33 ± 0.70 mm) by 21.8% of the sites, remained unchanged by 41.4% and increased (1.40 ± 0.78 mm) by 36.8%. Distopalatal FI (p < .001, odds ratio [OR]: 0.252, 95% confidence interval [CI] for OR: 0.118-0.540), residual pockets (p < .001, OR: 0.503, 95% CI: 0.429-0.590) and TM Degrees I-III (Degree I: p = .002, OR: 0.765, 95% CI: 0.646-0.905; Degree II: p = .006, OR: 0.658, 95% CI: 0.489-0.886; Degree III: p = .023, OR: 0.398, 95% CI: 0.180-0.879) correlated significantly with increasing PPD. CONCLUSIONS: Over 75% of PPD remained unchanged or increased during SPC. Distopalatal FI, TM Degrees I-III and residual pockets after APT lead to worsening of periodontal pockets.

Analysis of RAS and drug induced homo- and heterodimerization of RAF and KSR1 proteins in living cells using split Nanoluc luciferase.

The dimerization of RAF kinases represents a key event in their activation cycle and in RAS/ERK pathway activation. Genetic, biochemical and structural approaches provided key insights into this process defining RAF signaling output and the clinical efficacy of RAF inhibitors (RAFi). However, methods reporting the dynamics of RAF dimerization in living cells and in real time are still in their infancy. Recently, split luciferase systems have been developed for the detection of protein-protein-interactions (PPIs), incl. proof-of-concept studies demonstrating the heterodimerization of the BRAF and RAF1 isoforms. Due to their small size, the Nanoluc luciferase moieties LgBiT and SmBiT, which reconstitute a light emitting holoenzyme upon fusion partner promoted interaction, appear as well-suited to study RAF dimerization. Here, we provide an extensive analysis of the suitability of the Nanoluc system to study the homo- and heterodimerization of BRAF, RAF1 and the related KSR1 pseudokinase. We show that KRASG12V promotes the homo- and heterodimerization of BRAF, while considerable KSR1 homo- and KSR1/BRAF heterodimerization already occurs in the absence of this active GTPase and requires a salt bridge between the CC-SAM domain of KSR1 and the BRAF-specific region. We demonstrate that loss-of-function mutations impairing key steps of the RAF activation cycle can be used as calibrators to gauge the dynamics of heterodimerization. This approach identified the RAS-binding domains and the C-terminal 14-3-3 binding motifs as particularly critical for the reconstitution of RAF mediated LgBiT/SmBiT reconstitution, while the dimer interface was less important for dimerization but essential for downstream signaling. We show for the first time that BRAFV600E, the most common BRAF oncoprotein whose dimerization status is controversially portrayed in the literature, forms homodimers in living cells more efficiently than its wildtype counterpart. Of note, Nanoluc activity reconstituted by BRAFV600E homodimers is highly sensitive to the paradox-breaking RAFi PLX8394, indicating a dynamic and specific PPI. We report the effects of eleven ERK pathway inhibitors on RAF dimerization, incl. third-generation compounds that are less-defined in terms of their dimer promoting abilities. We identify Naporafenib as a potent and long-lasting dimerizer and show that the split Nanoluc approach discriminates between type I, I1/2 and II RAFi. Video Abstract.

What we see is what we touch? Sex estimation on the pelvis in virtual anthropology.

BACKGROUND: Computed tomography (CT) scans are a convenient means to study 3D reconstructions of bones. However, errors associated with the different nature of the observation, e.g. visual and tactile (on dry bone) versus visual only (on a screen) have not been thoroughly investigated. MATERIALS AND METHODS: We quantified the errors between modalities for sex estimation protocols of nonmetric (categorical and ordinal) and metric data, using 200 dry pelves of archaeological origin and the CT reconstructions of the same bones. In addition, we 3D surface scanned a subsample of 39 pelves to compare observations with dry bone and CT data. We did not focus on the sex estimation accuracy but solely on the consistency of the scoring, hence, the interchangeability of the modalities. RESULTS: Metric data yielded the most consistent results. Among the nonmetric protocols, ordinal data performed better than categorical data. We applied a slightly modified description for the trait with the highest errors and grouped the traits according to consistency and availability in good, intermediate, and poor. DISCUSSION: The investigated modalities were interchangeable as long as the trait definition was not arbitrary. Dry bone (gold standard) performed well, and CT and 3D surface scans performed better. We recommend researchers test their affinity for using virtual modalities. Future studies could use our consistency analysis and combine the best traits, validating their accuracy on various modalities.

Activation loop phosphorylation regulates B-Raf in vivo and transformation by B-Raf mutants.

Despite being mutated in cancer and RASopathies, the role of the activation segment (AS) has not been addressed for B-Raf signaling in vivo. Here, we generated a conditional knock-in mouse allowing the expression of the B-Raf(AVKA) mutant in which the AS phosphoacceptor sites T599 and S602 are replaced by alanine residues. Surprisingly, despite producing a kinase-impaired protein, the Braf(AVKA) allele does not phenocopy the lethality of Braf-knockout or paradoxically acting knock-in alleles. However, Braf(AVKA) mice display abnormalities in the hematopoietic system, a distinct facial morphology, reduced ERK pathway activity in the brain, and an abnormal gait. This phenotype suggests that maximum B-Raf activity is required for the proper development, function, and maintenance of certain cell populations. By establishing conditional murine embryonic fibroblast cultures, we further show that MEK/ERK phosphorylation and the immediate early gene response toward growth factors are impaired in the presence of B-Raf(AVKA). Importantly, alanine substitution of T599/S602 impairs the transformation potential of oncogenic non-V600E B-Raf mutants and a fusion protein, suggesting that blocking their phosphorylation could represent an alternative strategy to ATP-competitive inhibitors.

Smoking susceptibility as a predictive measure of cigarette and e-cigarette use among early adolescents.

OBJECTIVE: To assess the validity of a standard measure of smoking susceptibility for predicting cigarette and e-cigarette use in a sample of early adolescents in Argentina and Mexico. MATERIALS AND METHODS: A school-based longitudinal survey was conducted in 2014-16 among secondary students. We analyzed students who were never smokers of regular cigarettes or e-cigarettes at baseline and who completed both surveys. The main independent variable was smoking susceptibility. Multilevel logistic regression models were used, adjusting for sociodemographic and personal variables, social network use of cigarettes and exposure to advertising. RESULTS: In the adjusted analysis, smoking susceptibility independently predicted cigarette initiation (Argentina: AOR 2.28; 95%CI 1.66-3.14; Mexico: AOR 2.07; 95%CI 1.74-2.45) and current smoking (Argentina: AOR 3.61; 95%CI 2.48-5.24; Mexico: AOR 1.69; 95%CI 1.29-2.22); however, it only predicted e-cigarette initiation in Mexico (Mexico: AOR 1.29; 95%CI 1.02-1.63). CONCLUSIONS: Smoking susceptibility was a valid measure to predict future cigarette smoking in this sample.

Smoking in Movies and Adolescent Smoking Initiation: A Longitudinal Study among Argentinian Adolescents.

OBJECTIVE: To assess whether exposure to movie smoking is associated with cigarette smoking among Argentinian adolescents. STUDY DESIGN: A school-based longitudinal study involving 33 secondary schools in Argentina was performed. The sample included 2502 never smokers (average age at entry = 12.5 years), 1700 (67.9%) of whom completed follow-up surveys 17 months later. Exposure to the top 100 highest-grossing films for each year between 2009 and 2013 was assessed by content-coding films for tobacco and then by asking adolescents whether they had seen each of 50 titles, randomly selected from the larger pool, then parsing exposure into tertiles. Logistic regression models estimated aOR for the following transitions: nonsusceptible to susceptible never smoker, never smoker to ever smoker, and never smoker to current smoking (last 30 days). RESULTS: At follow-up, 34.4% of nonsusceptible never smokers became susceptible, 24.1% reported having tried smoking, and 9.4% were current smokers. Most exposure to movie smoking was from US-produced films (average 60.3 minutes compared with only 3.4 minutes from Argentine films). Greater exposure to smoking in movies was significantly associated with increased odds of becoming susceptible (aORfirst vs third tertile 1.77, 95% CI 1.30-2.41), of trying smoking (aORfirst vs third tertile 1.54, 1.14-2.08), and marginally associated with current smoking (AORfirst vs third tertile 1.54, 0.99-2.40). Exposure to smoking in US- or Argentine-produced films had similar associations. CONCLUSION: In Argentina, exposure to smoking in the movies predicted future smoking transitions among early adolescents, with most exposure coming from viewing US movies.

Time trends for tobacco and alcohol use in youth-rated films popular in Mexico and Argentina, from 2004-2012.

OBJECTIVE:: To examine and compare overall prevalence and time trends in tobacco and alcohol portrayals and brand appearances in youth-rated US and nationally-produced films that were the most successful in Argentina and Mexico from 2004-2012. MATERIALS AND METHODS:: Top-grossing nationally produced films from Argentina (n=73), Mexico (n=85) and the US (n=643) were content analyzed. Logistic regression was used to determine differences between Mexican, Argentine and US produced films. Linear regression models assessed significant cross-country differences in the mean number of tobacco and alcohol seconds. RESULTS:: Films from Mexico and Argentina were more likely than US films to contain tobacco, (OR=4.2; p<0.001) and (OR=7.2; p<0.001). Alcohol was present in 93% of Argentine, 83% in Mexican and 83% US films. CONCLUSIONS:: Smoking and alcohol were highly prevalent in nationally produced films. They may have a significant impact and should be targeted by policies to reduce youth exposure to portrayals of risk behaviors.

Distinct requirement for an intact dimer interface in wild-type, V600E and kinase-dead B-Raf signalling.

The dimerisation of Raf kinases involves a central cluster within the kinase domain, the dimer interface (DIF). Yet, the importance of the DIF for the signalling potential of wild-type B-Raf (B-Raf(wt)) and its oncogenic counterparts remains unknown. Here, we show that the DIF plays a pivotal role for the activity of B-Raf(wt) and several of its gain-of-function (g-o-f) mutants. In contrast, the B-Raf(V600E), B-Raf(insT) and B-Raf(G469A) oncoproteins are remarkably resistant to mutations in the DIF. However, compared with B-Raf(wt), B-Raf(V600E) displays extended protomer contacts, increased homodimerisation and incorporation into larger protein complexes. In contrast, B-Raf(wt) and Raf-1(wt) mediated signalling triggered by oncogenic Ras as well as the paradoxical activation of Raf-1 by kinase-inactivated B-Raf require an intact DIF. Surprisingly, the B-Raf DIF is not required for dimerisation between Raf-1 and B-Raf, which was inactivated by the D594A mutation, sorafenib or PLX4720. This suggests that paradoxical MEK/ERK activation represents a two-step mechanism consisting of dimerisation and DIF-dependent transactivation. Our data further implicate the Raf DIF as a potential target against Ras-driven Raf-mediated (paradoxical) ERK activation.

Assessing tobacco marketing receptivity among youth: integrating point of sale marketing, cigarette package branding and branded merchandise.

BACKGROUND: As countries prohibit tobacco marketing through traditional channels, marketing at point of sale (PoS) and through tobacco packaging is increasingly important for promoting tobacco consumption. OBJECTIVES: Assess the validity of a novel marketing receptivity index that considers frequency of PoS exposures, tobacco brand recall and ownership of branded merchandise. METHODS: Data come from a cross-sectional survey of 3172 secondary school students in Argentina. Questions assessed frequency of going to stores where tobacco is often sold; cued recall of brand names for 3 cigarette packages with brand name removed and ownership of branded merchandise. A four-level marketing receptivity index was derived: low PoS exposure only; high PoS exposure or recall of 1 brand; recall of 2 or more brands; and ownership of branded merchandise. Indicators of marketing receptivity and smoking involvement were regressed on the index, including in adjusted models that controlled for sociodemographics, social influences and sensation seeking. FINDINGS: Among never-smokers, the index had independent positive associations with smoking susceptibility (ie, adjusted OR (AOR)2v1=1.66; AOR3v1=1.64; AOR4v1=2.95), willingness to try a specific brand (ie, AOR2v1=1.45; AOR3v1=2.38; AOR4v1=2.20) and positive smoking expectancies (ie, Badj 2v1=0.09; Badj 3v1=0.18; Badj 4v1=0.34). A more marked dose-response independent association was found with current smoking behaviour (ie, AOR2v1=2.47; AOR3v1=3.16; AOR4v1=3.62). CONCLUSIONS: The marketing receptivity index was associated with important variation in smoking-related perceptions, intentions and behaviour among Argentine adolescents. Future research should determine the predictive validity and generalisability of this measure to other contexts, including the explanatory power gained by integrating cigarette package brand recognition tasks.

Piggybacking as a media advocacy strategy to increase enrolments in a gender-oriented smoking cessation programme.

Argentina is probably moving to the third phase of the smoking epidemic. Female smoking prevalence is expected to increase over the coming years. In Argentina, smoking cessation programmes usually do not provide specific treatment tailored to women. We implemented a 'piggybacking' media strategy with the goal of announcing the opening of the first gender-oriented smoking cessation programme in Argentina. Piggybacking is a well-known media advocacy strategy in which the newsworthiness of a particular story is increased by releasing it at the same time as a breaking news story about a related topic. We prepared a press release/report about tobacco use among women, as well as our gender-oriented clinic, for the local news media, which appeared in print around the time a well-known young Argentinean actress died. To assess the impact of this strategy, we reviewed media coverage after the press release was issued. We also compared the number of new participants in our programme during the 4 months before and after the report's publication. During the 4 months following our press release, we found five reports in print media, gave 22 radio and seven television interviews, and found 30 digital media publications drawing on our press release. When comparing the 4 months before with the 4 months after the strategy, new participants in our programme increased by 246.15%. This strategy could be a suitable alternative to other media advocacy strategies to increase the number of new participants in smoking cessation programmes.

Strong negative feedback from Erk to Raf confers robustness to MAPK signalling.

Protein levels within signal transduction pathways vary strongly from cell to cell. Here, we analysed how signalling pathways can still process information quantitatively despite strong heterogeneity in protein levels. We systematically perturbed the protein levels of Erk, the terminal kinase in the MAPK signalling pathway in a panel of human cell lines. We found that the steady-state phosphorylation of Erk is very robust against perturbations of Erk protein level. Although a multitude of mechanisms exist that may provide robustness against fluctuating protein levels, we found that one single feedback from Erk to Raf-1 accounts for the observed robustness. Surprisingly, robustness is provided through a fast post-translational mechanism although variation of Erk levels occurs on a timescale of days.

Biochemical predictors of cardiac rhythm at 1 year follow-up in patients with non-valvular atrial fibrillation.

C-reactive protein (CRP) is increased in patients with atrial fibrillation (AF) and it might predict the lack of sinus rhythm maintenance in the long term follow-up. Scarce data have been reported relating endothelial dysfunction and/or haemostatic or platelet markers to sinus rhythm maintenance in AF patients. We aimed to determine whether some biochemical markers including soluble thrombomodulin (sTM), soluble P-selectin, thrombin-antithrombin (TAT) complex and CRP levels could predict sinus rhythm maintenance at 1 year follow-up in non-valvular AF patients. 130 patients (70 males, mean age 67 ± 13 years) with newly diagnosed AF naïve of antithrombotic or antiplatelet therapy were studied. Baseline CRP, P-selectin, sTM and TAT levels were compared to those of 20 matched-healthy subjects in sinus rhythm. AF patients had significantly higher plasma CRP (10.5 ± 2.2 vs 3.25 ± 0.3 mg/l, p = 0.001), P-selectin (219 ± 141 vs 126 ng/ml, p = 0.01), and TAT (54 ± 237 vs 2.7 ± 3.3 ng/l, p = 0.001) levels compared with controls. Soluble TM levels, although higher than controls, did not reach statistical significance. Multivariate regression analyses showed that elevated CRP (OR = 4.8, p = 0.02) and sTM (OR = 1.05, p = 0.04) were the only two predictors of lack of sinus rhythm at 1 year follow-up. An altered inflammatory, haemostatic, endothelial and platelet environment exists in newly diagnosed AF patients. CRP and sTM levels might be used as biochemical markers to predict the status of cardiac rhythm at 1 year follow-up in some AF patients.

[Clinical characteristics, complications and mortality in 506 patients with infective endocarditis and determinants of survival rate at 10 years]. / Endocarditis infecciosa: características clínicas, complicaciones y mortalidad en 506 pacientes y factores pronósticos de sobrevida a 10 años (1998-2008). Estudio cooperativo nacional en endocarditis infecciosa en Chile (ECNEI-2).

BACKGROUND: Rates of morbidity and mortality in Infective Endocarditis (IE) remain high and prognosis in this disease is still difficult and uncertain. AIM: To study IE in Chile in its active phase during inpatient hospital stay and long term survival rates. MATERIAL AND METHODS: Observational prospective national cohort study of 506 consecutive patients included between June 1,1998 and July 31, 2008, from 37 Chilean hospitals (secondary and tertiary centers) nationwide. RESULTS: The main findings were the presence of Rheumatic valve disease in 22.1 % of patients, a history of intravenous drug abuse (IVDA) only in 0.7%, the presence of Staphylococcus aureus in 29.2% of blood cultures, negative blood cultures in 33.2%, heart failure in 51.7% and native valve involvement in 86% of patients. Echocardiographic diagnosis was achieved in 94% of patients. Hospital mortality was 26.1% and its prognostics factors were persisting infection (Odds ratio (OR) 6.43, Confidence Interval (CI) 1.45-28.33%), failure of medical treatment and no surgical intervention (OR 48.8; CI 6.67-349.9). Five and 10 years survival rates were 75.6 and 48.6%, respectively. The significant prognostic factors for long term mortality, determined by multivariate analysis were the presence of diabetes, Staphylococcus aureus infection, sepsis, heart failure, renal failure and lack of surgical treatment during the IE episode. CONCLUSIONS: The microbiologic diagnosis of IE must be urgently improved in Chile. Mortality rates are still high (26.1%) partly because of a high incidence of negative blood cultures and the need for more surgical valve interventions during in-hospital period. Long term prognostic factors for mortality should be identified early to improve outcome.

Perimortem Skeletal Sharp Force Trauma: Detection Reliability on CT Data, Demographics and Anatomical Patterns from a Forensic Dataset.

The increasing importance of trauma analysis by means of postmortem computed tomography (PMCT) is insufficiently reflected in forensic curricula, nor are best practice manuals available. We attempt to detect sharp force bone lesions on PMCT of closed forensic cases with the aims of assessing errors and pointing out patterns in anatomical location and manner of death (MOD). We investigated 41 closed sharp force fatality cases, with available PMCT and forensic reports. Two observers with different radiological training assessed the lesions on PMCT scans (2D and 3D) for comparison with the reports. Between 3% (suicides) and 15.3% (homicides) of sharp force injuries caused visible bone lesions. While our observations were repeatable, each forensic investigation left a similar number of bone lesions undetected. Injury patterns differed between MOD, with thoracic bone lesions being most frequent overall. Soft tissue injury location varied between the MOD. Associations between MOD and age as well as number of injuries were significant. The detection of bone lesions on PMCT for untrained forensic specialists is challenging, curricula and pertinent manuals are desirable. With the low frequency of bone lesions compared to soft tissue injuries, we should be aware when analyzing decomposed bodies.

Markedly increased Rho-kinase activity in circulating leukocytes in patients with chronic heart failure.

BACKGROUND: The small guanosine triphosphatase Rho and its target Rho-kinase have significant roles in experimental remodeling and ventricular dysfunction, but no data are available on Rho-kinase activation in patients with heart failure (HF). We hypothesized that, in patients with chronic HF, Rho-kinase in circulating leukocytes is activated and related to left ventricular (LV) remodeling and dysfunction. METHODS: Accordingly, Rho-kinase activity, assessed by the levels of phosphorylated to total myosin light chain phosphatase 1 (MYPT1-P/T) in circulating leukocytes, and echocardiographic LV function data were compared between patients with HF New York Heart Association functional class II or III due to systolic dysfunction (n = 17), healthy controls (n = 17), and hypertensive patients without HF (n = 17). RESULTS: In the control subjects, mean MYPT1-P/T ratio was 1.2 ± 0.2 (it was similar in the hypertensive patients without HF), whereas in patients with HF, it was significantly increased by >100-fold (P < .001). Both MYPT1-P/T and log MYPT1-P/T ratios were inversely correlated with ejection fraction (r = -0.54, P < .03 and r = -0.86, P < .001, respectively). Furthermore, in patients with HF with LV end-diastolic diameter <60 mm, MYPT1-P/T ratio was 35.8 ± 18.1, whereas it was significantly higher in patients with LV diameter ≥60 mm (P < .05). CONCLUSIONS: Rho-Kinase activity is markedly increased in patients with stable chronic HF under optimal medical treatment, and it is associated with pathologic LV remodeling and systolic dysfunction. Mechanisms of Rho-kinase activation in patients with HF, its role in the progression of the disease, and the direct effect of Rho-kinase inhibition need further investigation.

[Transcultural adaptation and validation of the Self-efficacy/Temptations Scale in Argentina]. / Adaptación transcultural y validación de la Escala de Autoeficacia/Tentaciones en Argentina.

Self-efficacy is the confidence in one's own abilities to avoid smoking in situations that can trigger desire. It is considered an important predictor of relapses. In Argentina there is no valid instrument to evaluate it. The objective of this article was to adapt and validate a Spanish version (in local Argentinean language) of the Self-Efficacy/Temptation Scale. The transcultural adaptation of the scale was carried out: translation, review by a committee of experts, reverse translation, new review and test of the questionnaire. Reliability (reproducibility, stability and internal consistency) and validity were then evaluated. We performed a factor analysis of main components with Varimax rotation with data from a sample of 200 smokers. A culturally and linguistically adapted version of the instrument was obtained, with inter-observer reliability of Intraclass Correlation Coefficient (ICC) = 0.90 and stability of ICC = 0.77. The internal consistency analysis yielded a global Crombach a = 0.93 and 0.85 for versions of 20 and 9 items, respectively. In the domain of Positive Affections a = 0.82 and a = 0.89; Negative Affections a = 0.89 and a = 0.90 and for Craving a = 0.84 and a = 0.75, also respectively for the two versions. In the exploratory factor analysis, three factors were identified that explained 95% of the variance of the questionnaire in its extensive version and 69% in its short version. A reliable and valid Argentinean version of the Self-Efficacy/Temptation Scale was obtained, to be applied to smokers.

La autoeficacia es la confianza en las propias habilidades para evitar fumar en situaciones quepuedan desencadenar el deseo. Se la considera un importante predictor de recaídas. En Argentina no existe un instrumento válido que permita evaluarla. El objetivo del presente trabajo fue adaptar y validar una versión en español (Argentina) de la Escala de Autoeficacia/Tentaciones. Se llevó a cabo la adaptación transcultural de la escala: traducción, revisión por un comité de expertos, traducción inversa, nueva revisión y prueba del cuestionario. Luego se evaluaron la confiabilidad (reproducibilidad, estabilidad y consistencia interna) y la validez. Se realizó un análisis factorial de componentes principales con rotación Varimax con datos de una muestra de 200 pacientes fumadores. Se obtuvo una versión cultural y lingüísticamente adaptada del instrumento, con una confiabilidad interobservador evaluada con el Coeficiente de Correlación Intraclase (ICC) = 0.90 y estabilidad de ICC = 0.77. En el análisis de la consistencia interna se obtuvo un a de Crombach global de 0.93 y de 0.85 para las versiones de 20 y 9 ítems, respectivamente. En el dominio de afectos positivos a = 0.82 y a = 0.89; afectos negativos a = 0.89 y a = 0.90 y para deseo intenso de fumar (craving) a = 0.84 y a = 0.75, también respectivamente para las dos versiones. En el análisis factorial exploratorio se identificaron tres factores que explicaron el 95% de la varianza del cuestionario en su versión extensa y 69% en su versión corta. Se obtuvo una versión argentina de la Escala de Autoeficacia/Tentaciones confiable y válida para ser aplicada a fumadores.

High-resolution insertion-site analysis by linear amplification-mediated PCR (LAM-PCR).

Integrating vector systems used in clinical gene therapy have proven their therapeutic potential in the long-term correction of immunodeficiencies. The integration loci of such vectors in the cellular genome represent a molecular marker unique for each transduced cell and its clonal progeny. To gain insight into the physiology of gene-modified hematopoietic repopulation and vector-related influences on clonal contributions, we have previously introduced a technology--linear amplification-mediated (LAM) PCR--for detecting and sequencing unknown DNA flanking sequences down to the single cell level (Supplementary Note online). LAM-PCR analyses have enabled qualitative and quantitative measurements of the clonal kinetics of hematopoietic regeneration in gene transfer studies, and uncovered the clonal derivation of non-leukemogenic and leukemogenic insertional side effects in preclinical and clinical gene therapy studies. The reliability and robustness of this method results from the initial preamplification of the vector-genome junctions preceding nontarget DNA removal via magnetic selection. Subsequent steps are carried out on a semisolid streptavidin phase, including synthesis of double complementary strands, restriction digest, ligation of a linker cassette onto the genomic end of the fragment and exponential PCR(s) with vector- and linker cassette-specific primers. LAM-PCR can be adjusted to all unknown DNA sequences adjacent to a known DNA sequence. Here we describe the use of LAM-PCR analyses to identify 5' long terminal repeat (LTR) retroviral vector adjacent genomic sequences.