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El propósito de la presente investigación fue establecer las propiedades psicométricas de una escala para medir actitudes en torno a las brechas generacionales en el trabajo. Dicha escala fue desarrollada, validada y aplicada a una muestra de 200 empleados puertorriqueños del área de la salud. La muestra tuvo una distribución equitativa por género (50%), aunque el 57% de la muestra eran enfermeros y la media de edad fue de 42 años (con una desviación estándar de 13.74). Inicialmente fueron desarrollados un total de 37 reactivos, de los cuales quedaron 16 después de pasar una validez de contenido por jueces expertos. Esta puntuación de validez de contenido (Índice de validez de contenido) fue de .99, lo que indicó que la escala, en efecto, tiene validez de contenido. Un estudio piloto (n=40) fue llevado a cabo para evaluar las instrucciones de la escala, la redacción de los reactivos y la operacionalización de la misma. Un análisis de discriminación (con “correlación ítem-total”) también fue llevado a cabo, dejando 10 reactivos. Por último, una validez de constructo con análisis de factores (utilizando la técnica de componentes principales, con rotación “varimax”) fue realizada, quedando finalmente dos factores en la escala. La confiabilidad de Alpha de Cronback obtenida fue de .83. Los resultados indican que la escala tiene validez de contenido, de constructo, y que es confiable. Los resultados dan cuenta de que la validez y fiabilidad de la escala para medir actitudes generacionales en el trabajo, aunque se recomienda realizar más estudios con la escala y otros estudios de validez para aumentar la fiabilidad de la prueba.

Introducción: el síndrome metabólico (SM) se considera una epidemia mundial; el aumento global de su prevalencia se ha extendido en los países tanto industrializados como en vías de desarrollo, como resultado de una mayor proporción de obesidad y sedentarismo en la población. El objetivo de esta investigación fue determinar la prevalencia de SM en conductores de transporte intermunicipal de Tunja en 2017. Materiales y métodos: se realizó un estudio analítico de corte transversal a una muestra de 115 conductores de transporte intermunicipal en la ciudad de Tunja, se evaluaron índices antropométricos, glicemia, triglicéridos, HDL y se aplicó el cuestionario internacional de actividad física IPAQ. La presencia de SM se identificó de acuerdo con los criterios de NCEP/ATP III, se calculó la razón de prevalencias y se determinó significancia mediante la prueba exacta de Fisher y de U de Mann-Whitney. Resultados: se encontró una prevalencia de SM del 10.4 % (12/115; ic 95 % 4.6-15.7) y se hallaron diferencias significativas entre los pacientes con y sin SM en relación con la hipertensión (p = 0.03), niveles de glucosa (p = 0.0004), triglicéridos (p =

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Oral anticoagulation prevents strokes in patients with atrial fibrillation but, for reasons that remain unclear, less than 40% of all patients with atrial fibrillation receive warfarin. The literature postulates that patient and clinician preferences may explain this low utilization.~Background~Background~The proposed research seeks to answer the following questions: i) When assessed systematically, do patients' and clinicians' preferences explain the utilization of warfarin to prevent strokes associated with atrial fibrillation? ii) To what extent do patients' and clinicians' treatment preferences differ? iii) What factors explain any differences that exist in treatment preferences between patients and clinicians? To answer these questions we will conduct a two-phase study of patient and clinician preferences for health states and treatments. In the first phase of this study we will conduct structured interviews to determine their treatment preferences for warfarin vs. aspirin to prevent strokes associated with atrial fibrillation using the probability trade-off technique. In the same interview, we will conduct preference-elicitation exercises using the feeling thermometer to identify the utilities that patients place on taking medication (warfarin and aspirin), and on having a mild stroke, a severe stroke, and a major bleed. In the second phase of the study we will convene focus groups of clinicians and patients to explore their answers to the exercises in the first phase.~Design~Methods~This is a study of patient and clinician preferences for health states and treatments. Because of its clinical importance and our previous work in this area, we will conduct our study in the clinical context of the decision to use antithrombotic agents to reduce the risk of stroke in patients with non-valvular chronic atrial fibrillation.~Discussion~Conclusions [ABSTRACT FROM AUTHOR]

Introduction and Objectives:  The ureteral catheter is used to provide adequate drainage of the upper urinary tract. They have been used to prevent or relieve ureteral obstruction due to multiple causes. The literature mentions the standard placement of double J catheter guided by fluoroscopy, reporting the use of ultrasound exclusively in cases of pregnant patients in order to avoid ionizing radiation. Based on the aforementioned, the primary objective was to evaluate the placement of double J catheter guided by ultrasound, as an alter native technique to the use of the fluoroscope in ureteral obstructions, as secondary objectives the intensity of the pain and the complications of the procedure., Materials and Methods: An observational, descriptive and prospective study was carried out with 41patients who attended the lithiasis consultation., Results: A total of 41 patients who under went double J catheter guided by ultrasound were evaluated, achieving it successfully in 35 of the cases, resulting in bad position in 2 patients and being impossible to insert it in 4. Averageage was 41 years, size of the lithium 10.41mm. The complications presented were 17% with the use of the scale of Clavien modified these complications were represented as follows: 4.9% grade I back pain, 7.2% grade II lower urinary tract symptoms, lower tract infections, and 4.9% grade IIIa malposition., Conclusions: The placement of ureteral catheters guided by ultrasound, represent a useful tool, safe, with manageable complications, taking place in times adjusted to the procedure.

We study entropy production in nanoscale devices, which are coupled to finite heat baths. This situation is of growing experimental relevance, but most theoretical approaches rely on a formulation of the second law valid only for infinite baths. We fix this problem by pointing out that Clausius' paper from 1865 already contains an adequate formulation of the second law for finite heat baths, which can be also rigorously derived from a microscopic quantum description. This Clausius inequality shows that nonequilibrium processes are less irreversible than previously thought. We use it to correctly extend Landauer's principle to finite baths and we demonstrate that any heat engine in contact with finite baths has a higher efficiency than previously thought. Importantly, our results are easy to study, requiring only the knowledge of the average bath energy.

The plastid-encoded genes of higher plants are transcribed by at least two types of RNA polymerases, the nuclear-encoded RNA polymerase (NEP) and the plastid-encoded RNA polymerase (PEP). In mature photosynthesizing leaves, the vast majority of the genes are transcribed by PEP. However, the regulatory mechanisms controlling plastid transcription during early light response is unclear. Chloroplast development is suggested to be associated with a shift in the usage of the primary RNA polymerase from NEP to PEP as the expression of the plastid-encoded photosynthesis genes is induced upon light exposure. Assembly of the PEP complex has been suggested as a rate-limiting step for full activation of plastid-encoded photosynthesis gene expression. However, two sigma factor mutants, sig2 and sig6, with reduced PEP activity, showed significantly lower expression of the plastid-encoded photosynthesis genes already in the dark and during the first hours of light exposure indicating that PEP activity is required for basal expression of plastid-encoded photosynthesis genes in the dark and during early light response. Furthermore, in etioplasts and proplastids a fully assembled PEP complex was revealed on Blue Native PAGE. Our results indicate that a full assembly of the PEP complex is possible in the dark and that PEP drives basal transcriptional activity of plastid-encoded photosynthesis genes in the dark. Assembly of the complex is most likely not a rate-limiting step for full activation of plastid-encoded photosynthesis gene expression which is rather achieved either by the abundance of the PEP complex or by some posttranslational regulation of the individual PEP components., (© 2020 Scandinavian Plant Physiology Society.)

In the present work, a strain of the basidiomycete fungus Trametes polyzona was used to decolorize the Amaranth dye. The decolorization was carried out in an Airlift reactor with three flow regimes: 1, 2, and 3 vvm. The results showed that the decolorization was a function of the flow regime. The decolorization times for the regimes of 1, 2, and 3 vvm were 30, 25, and 19 days, respectively. The COD (Chemical Oxygen Demand) decreased from 1600 to 72 mg COD/L. The enzymatic activity kinetics of laccase (Lcc), lignin peroxidase (LiP), and manganese peroxidase (MnP) were determined. In all the treatments, the enzyme LiP was expressed during the first 6 days, at which point 80% decolorization was observed, whereas Lcc and MnP enzymes were produced from day 6 until the end of the decolorization process. The effluent generated showed no inhibitory effects on the growth of the algae Nannochloropsis salina . T. polyzona showed great versatility in the decolorization of synthetic effluents containing the Amaranth dye, and the fungus was able to use this dye as its only carbon source starting at the beginning of the process. LiP was the enzyme that contributed the most to the decolorization process, and on average, 95% decreases in color and the COD were observed., Competing Interests: The authors declare no conflict of interest., (© 2020 Published by Elsevier Ltd.)

The two-point measurement scheme for computing the thermodynamic work performed on a system requires it to be initially in equilibrium. The Margenau-Hill scheme, among others, extends the previous approach to allow for a non-equilibrium initial state. We establish a quantitative comparison between both schemes in terms of the amount of coherence present in the initial state of the system, as quantified by the l1-coherence measure. We show that the difference between the two first moments of work, the variances of work, and the average entropy production obtained in both schemes can be cast in terms of such initial coherence. Moreover, we prove that the average entropy production can take negative values in the Margenau-Hill framework.

In this work was to evaluate the conidiospores production of Trichoderma harzianum using barley straw as substrate. Four growth conditions were used; washed and unwashed barley straw and washed and unwashed barley straw supplemented with mineral salts. The highest spore production was observed when washed barley straw supplemented with mineral salts with 1.56 × 10 10 conidiospores/gram of dry matter (gdm) at 216 h of cultivation was used. The effect of substrate moisture on spore production was studied, three initial moisture levels of the substrate were tested and it was observed that a humidity of 80 % of the substrate improves the production of conidiospores reaching a concentration of 2.35 × 10 10 conidiospores/gdm at 136 h. Finally, conidiospores viability was evaluated for 12 months by keeping them on the conidia and substrate, and viability of 71 % of the conidiospores was observed, so this maintenance method is an excellent means of conserving the conidiospores viability., Competing Interests: The authors declare that they have no conflict of interest., (© 2020 The Authors.)

It has been postulated that monitoring measurable residual disease (MRD) could be used as a surrogate marker of progression-free survival (PFS) in chronic lymphocytic leukemia (CLL) patients after treatment with immunochemotherapy regimens. In this study, we analyzed the outcome of 84 patients at 3 years of follow-up after first-line treatment with fludarabine, cyclophosphamide and rituximab (FCR) induction followed by 36 months of rituximab maintenance thearpy. MRD was assessed by a quantitative four-color flow cytometry panel with a sensitivity level of 10 -4 Eighty out of 84 evaluable patients (95.2%) achieved at least a partial response or better at the end of induction. After clinical evaluation, 74 patients went into rituximab maintenance and the primary endpoint was assessed in the final analysis at 3 years of follow-up. Bone marrow (BM) MRD analysis was performed after the last planned induction course and every 6 months in cases with detectable residual disease during the 36 months of maintenance therapy. Thirty-seven patients (44%) did not have detectable residual disease in the BM prior to maintenance therapy. Interestingly, 29 patients with detectable residual disease in the BM after induction no longer had detectable disease in the BM following maintenance therapy. After a median followup of 6.30 years, the median overall survival (OS) and PFS had not been reached in patients with either undetectable or detectable residual disease in the BM, who had achieved a complete response at the time of starting maintenance therapy. Interestingly, univariate analysis showed that after rituximab maintenance OS was not affected by IGHV status (mutated vs unmutated OS: 85.7% alive at 7.2 years vs 79.6% alive at 7.3 years, respectively). As per protocol, 15 patients (17.8%), who achieved a complete response and undetectable peripheral blood and BM residual disease after four courses of induction, were allowed to stop fludarabine and cyclophosphamide and complete two additional courses of rituximab and continue with maintenance therapy for 18 cycles. Surprisingly, the outcome in this population was similar to that observed in patients who received the full six cycles of the induction regimen. These data show that, compared to historic controls, patients treated with FCR followed by rituximab maintenance have high-quality responses with fewer relapses and improved OS. The tolerability of this regime is favorable. Furthermore, attaining an early undetectable residual disease status could shorten the duration of chemoimmunotherapy, reducing toxicities and preventing long-term side effects. The analysis of BM MRD after fludarabine-based induction could be a powerful predictor of post-maintenance outcomes in patients with CLL undergoing rituximab maintenance and could be a valuable tool to identify patients at high risk of relapse, influencing further treatment strategies. This trial is registered with EudraCT n. 2007-002733-36 and ClinicalTrials.gov Identifier: NCT00545714., (Copyright© 2019 Ferrata Storti Foundation.)

Although many sulfur containing garlic compounds present antioxidant activity, little is known about molecular mechanisms through which these compounds react with reactive oxygen species. In this work, the reactivity and the hydrogen peroxide scavenger reaction mechanisms (including thermodynamics and kinetics aspects) of allyl methyl disulfide in aqueous phase are studied employing density functional theory computational methods. Three reactive sites susceptible for electrophilic attack are found over sulfur atoms and the double bond allyl moiety. For each detected site, one redox reaction is proposed and analyzed. All reactions are thermodynamically feasible, whereas attack over the methyl bound sulfur atom is kinetically favored.

Activation of the plastid-encoded RNA polymerase is tightly controlled and involves a network of phosphorylation and, as yet unidentified, thiol-mediated events. Here, we characterize PLASTID REDOX INSENSITIVE2, a redox-regulated protein required for full PEP-driven transcription. PRIN2 dimers can be reduced into the active monomeric form by thioredoxins through reduction of a disulfide bond. Exposure to light increases the ratio between the monomeric and dimeric forms of PRIN2. Complementation of prin2-2 with different PRIN2 protein variants demonstrates that the monomer is required for light-activated PEP-dependent transcription and that expression of the nuclear-encoded photosynthesis genes is linked to the activity of PEP. Activation of PEP during chloroplast development likely is the source of a retrograde signal that promotes nuclear LHCB expression. Thus, regulation of PRIN2 is the thiol-mediated mechanism required for full PEP activity, with PRIN2 monomerization via reduction by TRXs providing a mechanistic link between photosynthetic electron transport and activation of photosynthetic gene expression.

The GAUDI study assessed safety and preliminary efficacy of induction therapy with obinutuzumab plus chemotherapy, followed by maintenance therapy with obinutuzumab alone, in previously untreated patients with follicular lymphoma. Assignment to chemotherapy was decided on a per-center basis before the patients' enrollment. Patients (n=81) received four to six cycles of obinutuzumab plus bendamustine every 4 weeks or six to eight cycles of obinutuzumab plus CHOP every 3 weeks. Patients with an end-of-treatment response were eligible for obinutuzumab maintenance therapy every 3 months for 2 years or until disease progression. Induction treatment was completed by 90% of patients in the obinutuzumab plus bendamustine group and 95% in the obinutuzumab plus CHOP group, while maintenance was completed by 81% and 72% of patients, respectively. All patients experienced at least one adverse event during induction, most commonly infusion-related reactions (58%), the majority of which were grade 1/2. The most common hematologic adverse event was grade 3/4 neutropenia (36% during induction and 7% during maintenance). One treatment-related death occurred during the maintenance phase. At the end of induction, 94% of patients had achieved an overall response, with complete response based on computed tomography in 36%. The progression-free survival rate at 36 months was 90% in the obinutuzumab plus bendamustine group and 84% in the obinutuzumab plus CHOP group. These results demonstrate that induction therapy with obinutuzumab plus bendamustine or obinutuzumab plus CHOP, followed by obinutuzumab maintenance, is associated with tolerable safety and promising efficacy. This study is registered at ClinicalTrials.gov as NCT00825149., (Copyright© Ferrata Storti Foundation.)

Background  Kaposiform hemangioendothelioma (KHE) is a vascular tumor frequently associated with Kasabach-Merritt phenomenon (KMP), characterized by severe thrombocytopenia and consumptive coagulopathy. Visceral involvement in KHE is rare. In our recent experience, sirolimus has shown to be an effective treatment in cutaneous KHE, becoming indeed the treatment of choice in KMP. We report a case of pancreatic KHE associated with KMP and refractory to sirolimus. Case Report  A 4-month-old infant is referred for obstructive jaundice (10 mg/dL conjugated bilirubin) secondary to vascular pancreatic tumor. Magnetic resonance (MR) and immunohistochemistry were compatible with KHE, but the tumor was considered unresectable. We initiated sirolimus (0.8 mg/m 2 /12 h) to treat KMP, and interventional radiology was performed for percutaneous biliary diversion. This procedure prompted KMP (platelets: 51,000/µL). Sirolimus treatment for 7 days showed no effect; therefore, we started our VAT protocol (vincristine/aspirine/ticlopidin) with great response after 10 days (platelets: 3,70,000/µL). Three months later, percutaneous biliary diversion was replaced by a biliary stent. The tumor disappeared leaving fibrosis and dilatation of biliary tract needing hepaticojejunostomy 6 months later. Discussion  It is difficult to establish protocols for an unusual presentation of a tumor with different targets. This is a reason collaborative multicenter studies should be performed. Management of obstructive jaundice secondary to a tumor that usually regresses in 10 years is an added challenge; therefore, the management should be led by a multidisciplinary team. Sirolimus treatment in cutaneous KHE has been described as successful in the literature, as well as in our own experience; however, it failed in our first patient with visceral KHE. We need to investigate the different response to pharmacological agents in tumors with similar histopathology, but with visceral involvement.

Objectives: The aim of this study was to analyze the usefulness of physical examination, C-reactive protein (CRP), procalcitonin (PCT), white blood cell (WBC) count, and absolute neutrophils counts (ANCs) for the diagnosis of invasive bacterial infections (IBIs) and potentially serious bacterial infections in infants younger than the age of 3 months presenting with fever without source (FWS) to the emergency department (ED)., Methods: A descriptive retrospective study that includes all infants aged younger than 3 months who presented with FWS to the ED between July 2008 and January 2012. We evaluated diagnostic performance for each test by receiver operating characteristic curve analysis. Sensitivity, specificity, positive likelihood ratio, and negative likelihood ratio were also calculated., Results: Three hundred eighteen patients met the inclusion criteria. Eleven bacteremia (3.5%) and 76 urinary tract infections (23.9%) were diagnosed. To detect IBI, the areas under the curve for the different tests were as follows: PCT, 0.77 (95% confidence interval [CI], 0.57-0.96); CRP, 0.54 (95% CI, 0.36-0.73); ANC, 0.53 (95% CI, 0.34-0.71); and WBC, 0.42 (0.24-0.61). To detect potentially serious bacterial infections, the areas under the curve were as follows: PCT, 0.66 (95% CI, 0.59-0.74); CRP, 0.68 (0.60-0.76); ANC, 0.64 (0.56-0.71); and WBC, 0.66 (0.58-0.72)., Conclusions: Procalcitonin is better than CRP, WBC, and ANC to confirm or dismiss the presence of an IBI in infants aged younger than 3 months presenting with FWS to the ED. However, it could not identify almost 30% of infants with IBI. Most patients diagnosed with IBI (10 of 11) presented abnormal values in at least one of the analytical parameters and/or physical appearance. Four of 5 patients with IBI and well appearing presented abnormal results in at least one of the analytical parameters. Therefore, the development of tools combining different tests including the new biomarkers could increase the reliability of the tests for the diagnosis of IBI in these patients.

Background: Exploration of values and preferences in the context of anticoagulation therapy for atrial fibrillation (AF) remains limited. To better characterize the distribution of patient and physician values and preferences relevant to decisions regarding anticoagulation in patients with AF, we conducted interviews with patients at risk of developing AF and physicians who manage patients with AF., Methods: We interviewed 96 outpatients and 96 physicians in a multicenter study and elicited the maximal increased risk of bleeding (threshold risk) that respondents would tolerate with warfarin vs. aspirin to achieve a reduction in three strokes in 100 patients over a 2-year period. We used the probabilistic version of the threshold technique., Results: The median threshold risk for both patients and physicians was 10 additional bleeds (10 P = 0.7). In both groups, we observed large variability in the threshold number of bleeds, with wider variability in patients than clinicians [patient range: 0-100, physician range: 0-50]. We observed one cluster of patients and physicians who would tolerate <10 bleeds and another cluster of patients, but not physicians, who would accept more than 35., Conclusions: Our findings suggest wide variability in patient and physician values and preferences regarding the trade-off between strokes and bleeds. Results suggest that in individual decision making, physician and patient values and preferences will often be discordant; this mandates tailoring treatment to the individual patient's preferences., (© 2014 John Wiley & Sons Ltd.)

Hypertension and obesity are two closely related pathologies in clinical practice. Currently, about one billion adults worldwide are overweight, and it is estimated that, if no serious action is taken to effect profound change, that figure will continue to rise throughout this century. Hypertension is also a serious public health problem worldwide, one that, along with type 2 diabetes, is growing due to increases in both life expectancy and obesity. However, the rate of increase varies by population group. For example, in the United States, the prevalence of obesity differs quite markedly among the white population of European origin, African American individuals, and the Latin American population. This disparity exists among other populations as well, such as that of Argentina, where obesity is less prevalent than in the United States. This significant difference between the Argentinean population and that of American whites may be explained by the former population's migratory origin and distinct eating habits.

Background: elementary school children spend little time at the school in Mexico. The aim was to investigate the activities after school time in children and to determine the presence of overweight and obesity., Methods: one hundred and seventy two children of both sexes were included. They belonged to the first and second grade of an official primary school from a marginalized zone. Somatometry was obtained and the type of physical practice and motifs for no practice physical activity were investigated. Descriptive statistics were applied., Results: the frequency of underweight was 12.1 %, overweight 19 %, and obesity 18.5 %. Sedentary activities like watching television and playing video games were the most common registered, whereas sports were the lowest physical activity practiced, especially in girls. The parents' lack of time, low family income, and the scarcity of safe spaces where children can play and practice physical activity or sports were the most related factors., Conclusions: poverty and insecurity of the neighborhoods influence the outside activities in after school time. The high frequency of underweight, overweight and obesity in school children were associated to the lack of physical activity.

The clinical heterogeneity that characterizes chronic lymphocytic leukemia (CLL), with survival times ranging from months to decades, reflects its biological diversity. Our understanding of the biology of CLL has helped us identify several markers of prognostic significance, by which CLL can be differentiated into several distinct diseases. The presence of specific chromosomal abnormalities is a prognostic indicator of disease progression and survival. Conventional cytogenetic analyses have revealed chromosomal aberrations in 40-50% of patients, but the detection of abnormalities is limited by the low mitotic activity of CLL cells. Metaphase analysis has recently undergone a "revival" because the metaphase yield has been improved by stimulation of CLL cells with alternative methods. Fluorescence in situ hybridization identifies chromosomal changes in approximately 80% of patients with CLL, and comparative genomic hybridization using high-density arrays (i.e., array comparative genomic hybridization [aCGH]) enables high-resolution genome-wide scanning for detecting copy number alterations in a single hybridization. The mutational status of the immunoglobulin heavy chain variable (IGHV) genes identifies two subsets of CLL with different outcomes. Unfortunately, the determination of IGHV mutation status may not be practical in all laboratories, and for this reason characteristics that are correlated with IGHV mutation status are needed-zeta-chain associated (TCR) protein kinase 70 kDa (ZAP-70) being that most commonly used currently in routine clinical practice. Whole genome sequencing has offered new insights into the mutational status of the disease, highlighting the role of several genes previously unrelated to CLL. Of these, NOTCH1 and SF3B1 are the most frequently mutated genes that predict poor prognosis. MicroRNA alterations are also involved in the initiation and progression of CLL, and the expression levels of some microRNAs correlate with previously established prognostic markers such as IGHV mutation status or ZAP-70. In addition, both global and gene-specific aberrant DNA methylation have been observed in CLL. Aberrant methylation has been described for genes that are specifically deregulated in CLL, such as BCL2, TCL1, and ZAP-70. Expanding knowledge of aberrant methylation profiles in CLL has a potential future impact on diagnosis, prognosis, and prediction of treatment response in CLL patients., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Epigenetic deregulation is considered a common hallmark of cancer. Nevertheless, recent publications have demonstrated its association with a large array of human diseases. Here, we explore the DNA methylation dynamics in blood samples during hematopoietic cell transplant and how they are affected by pathophysiological events during transplant evolution. We analyzed global DNA methylation in a cohort of 47 patients with allogenic transplant up to 12 months post-transplant. Recipients stably maintained the donor's global methylation levels after transplant. Nonetheless, global methylation is affected by chimerism status. Methylation analysis of promoters revealed that methylation in more than 200 genes is altered 1 month post-transplant when compared with non-pathological methylation levels in the donor. This number decreased by 6 months post-transplant. Finally, we analyzed methylation in IFN-γ, FASL, IL-10, and PRF1 and found association with the severity of the acute graft-versus-host disease. Our results provide strong evidence that methylation changes in blood are linked to underlying physiological events and demonstrate that DNA methylation analysis is a viable strategy for the study of transplantation and for development of biomarkers.

We have evaluated the use of CD138+ positively selected bone marrow samples to identify a molecular target for minimal residual disease assessment by polymerase chain reaction (PCR) in 25 untreated patients with multiple myeloma. A fraction of each sample was used for CD138+ selection, and the rest served as a reference control. VDJH, DJH, and Kde gene rearrangements were tested for amplification according to the BIOMED-2 Concerted Action. PCR products were directly sequenced in an automated ABI 3130 DNA sequencer using Big-Dye terminators. Within the CD138+ selected group, VDJH rearrangements were detected in all cases (100 %), DJH in 16 (64 %), and Kde in 18 (72 %) cases; whereas in the control samples, VDJH, DJH, and Kde rearrangements were detected in 19 (76 %), 11 (44 %), and 12 (48 %) cases, respectively. After sequencing, 24 (96 %) cases within the CD138+ group had a PCR target for MRD detection compared with 15 (60 %) cases in the control group. We conclude that the use of CD138+ positively selected bone marrow samples increases the applicability of minimal residual disease studies by PCR in patients with multiple myeloma.

Background and Objectives: Minimal residual disease (MRD) assessment by PCR in multiple myeloma (MM) has several shortcomings, including the lack of a suitable target. Kappa deleting element (KDE) rearrangements occur in virtually all Ig-lambda B-cell malignancies and in 1/3 of Ig-kappa are not affected by somatic hypermutation and, as in ALL, could be used as PCR targets., Methods: We have first investigated the incidence, gene segment usage, and CDR3 composition of IGK-KDE rearrangements in 96 untreated myeloma patients. Second, we tested 16 KDE gene rearrangements as molecular targets for MRD assessment by RQ-PCR using a germline reverse primer and a germline Taqman probe in combination with allele-specific oligonucleotides (ASO) as forward primers., Results: Monoclonal KDE rearrangements were amplified in 45% (43/96) of cases, monoallelic in 2/3 of them (29 cases), and biallellic in the remaining 14 cases. Overall, 88% of cases were successfully sequenced, KDE being equally frequently rearranged with VK and with intron-Recombination signal sequence (RSS). Median numbers of inserted and deleted nucleotides in the junctional region were one and five, respectively., Conclusions: Using KDE rearrangements as additional PCR target for MRD assessment in MM improves the applicability of these studies in 9% of cases overall and in 20% of lambda cases. Its use in the latter subset could represent a significant advance., (© 2012 John Wiley & Sons A/S.)

Background: Risk factors associated with monoclonal B-cell lymphocytosis (MBL), a potential precursor of chronic lymphocytic leukaemia (CLL), remain unknown., Methods: Using a cross-sectional study design, we investigated demographic, medical and behavioural risk factors associated with MBL. "Low-count" MBL (cases) were defined as individuals with very low median absolute count of clonal B-cells, identified from screening of healthy individuals and the remainder classified as controls. 452 individuals completed a questionnaire with their general practitioner, both blind to the MBL status of the subject. Odds ratios (OR) and 95% confidence interval (CI) for MBL were estimated by means of unconditional logistic regression adjusted for confounding factors., Results: MBL were detected in 72/452 subjects (16%). Increasing age was strongly associated with MBL (P-trend<0.001). MBL was significantly less common among individuals vaccinated against pneumococcal or influenza (OR 0.49, 95% confidence interval (CI): 0.25 to 0.95; P-value=0.03 and OR: 0.52, 95% CI: 0.29 to 0.93, P-value=0.03, respectively). Albeit based on small numbers, cases were more likely to report infectious diseases among their children, respiratory disease among their siblings and personal history of pneumonia and meningitis. No other distinguishing epidemiological features were identified except for family history of cancer and an inverse relationship with diabetes treatment. All associations described above were retained after restricting the analysis to CLL-like MBL., Conclusion: Overall, these findings suggest that exposure to infectious agents leading to serious clinical manifestations in the patient or its surroundings may trigger immune events leading to MBL. This exploratory study provides initial insights and directions for future research related to MBL, a potential precursor of chronic lymphocytic leukaemia. Further work is warranted to confirm these findings.

Analysis of short tandem repeats (STR) by PCR analysis is routinely used in chimerism diagnostics to monitor donor engraftment and to diagnose relapse. Some applications require chimerism analysis of low cell numbers, but no standardized protocol is available for DNA isolation from 1000 to 30,000 cells. The EU-supported EuroChimerism Consortium (project QLRT-2001-01485) selected four different protocols for 'small-scale' DNA isolation, which were tested by six laboratories for their ability to recover reproducible amounts of good quality DNA, suited for PCR-based STR analysis. The protocols included two direct lysis methods with and without detergents and proteinase K, and two commercial column-based kits. The direct lysis method using detergents and proteinase K showed the highest DNA recovery and the best performance in the multiplex PowerPlex16 STR assay. DNA isolated with this method also showed the highest sensitivity in chimerism analysis using singleplex PCR reactions of EuroChimerism STR markers. Sensitivity was reached ranging from 1 to 20% of recipient cells in a donor background. In conclusion, the direct lysis method using detergents and proteinase K is a standardized DNA isolation method well suited for chimerism studies on low cell numbers.