Novel alleles of the Transforming Growth Factor β-1 promoter and exon 1

Transforming growth factor β-1, encoded by the TGFB1 gene, is a cytokine that plays a central role in many physiologic and pathogenic processes, having pleiotropic effects on cell proliferation, differentiation, migration and survival. Regulatory activity for this gene has been demonstrated for approximately 3.0 kb between positions -2,665 and +423 from its translational start site. This region includes two promoter sites, two negative regulatory elements and two enhancers, in addition to part of the protein’s signal peptide. An important amount of polymorphism has been reported for this gene, including 17 promoter and exon 1 alleles defined on the basis or 18 polymorphisms . Polymorphism in TGFB1 has been associated to differential levels of expression of this cytokine and to genetic risk in cancer and hematopoietic stem cell (HSCT) and organ transplantation. In this report, we extend this list by presenting novel alleles formed by new polymorphic positions and a new combination of the previously described polymorphisms. These novel alleles have been found during the typing of a cohort of unrelated haematopoietic stem cell transplantation patient-donor pairs, and of healthy volunteer donors

High resolution HLA-A, -B, -C and -DRB1 allele and haplotype frequencies in the Costa Rica Central Valley population

The Costa Rica Central Valley population (CCVP) is the major population in this country, accounting for over 60% of the Costa Rican inhabitants concentrated since colonial times in a 2,500 km2 intermontane region. Interesting historic, demographic and genetic characteristics of this hybrid population have attracted researchers interested in testing genetic associations for various diseases. However, no study describing Human Leukocyte Antigen (HLA) frequencies by molecular methods had been performed. We have recently described low resolution HLA allele group and haplotype frequencies in a sample of this population. In this report, we extend our study to high resolution by sequence-based typing of exons 2, 3 and 4 for class I, and exon 2 for HLA-DRB1. DNA was extracted from blood or saliva samples from a cohort of 205 non-related healthy donors recruited as part of the University of Costa Rica’s Centre for Research in Hematology and Related Disorders (CIHATA) DNA bank. All participants were born in the CCVP and gave informed consent. A total of 37 HLA-A, 61 HLA-B, 24 HLA-C and 38 HLA-DRB1 alleles were seen in this sample. The five most frequent alleles for these genes are HLA-A*02:01:01, HLA-A*24:02:01, HLA-A*03:01:01, HLA-A*01:01:01, HLA-A*68:01:02, HLA-B*07:02:01, HLA-B*40:02:01, HLA-B*35:01:01, HLA-B*44:02:01, HLA-B*14:02:01, HLA-C*04:01:01, HLA-C*07:02:01, HLA-C*03:05, HLA-C*06:02:01, HLA-C*07:01:01, HLA-DRB1*13:01:01G, HLA-DRB1*04:07:01G, HLA-DRB1*15:01:01G, HLA-DRB1*03:01:01G, and HLA-DRB1*07:01:01G. Preliminary haplotype estimation results show, as a proxy for admixture proportions, that 68% of the extended haplotypes are Caucasian, while 23% are Amerindian in origin and 9% are clearly Sub-Saharan African. Principal coordinates analysis based on HLA-A and –B allele group frequencies reveals that this population clusters among other admixed groups with strong Caucasian component that lie closely to Iberian populations

Testing matter effects in propagation of atmospheric and long-baseline neutrinos

We quantify our current knowledge of the size and flavor structure of the matter effects in the evolution of atmospheric and long-baseline neutrinos based solely on the analysis of the corresponding neutrino data. To this aim we generalize the matter potential of the Standard Model by rescaling its strength, rotating it away from the e-e sector, and rephasing it with respect to the vacuum term. This phenomenological parametrization can be easily translated in terms of non-standard neutrino interactions in matter. We show that in the most general case, the strength of the potential cannot be determined solely by atmospheric and long-baseline data. However its flavor composition is very much constrained and the present determination of the neutrino masses and mixing is robust under its presence. We also present an update of the constraints arising from this analysis in the particular case in which no potential is present in the e-mu and e-tau sectors. Finally we quantify to what degree in this scenario it is possible to alleviate the tension between the oscillation results for neutrinos and antineutrinos in the MINOS experiment and show the relevance of the high energy part of the spectrum measured at MINOS.Comment: PDFLaTeX file using JHEP3 class, 25 pages, 7 figures included. Accepted for publication in JHE

Key determinants in building financial capability among middle schoolers with a school-based financial literacy education program

Rising in importance at various life stages, financial literacy and welfare-enhancing financial behaviors are crucial life-skills for youth to develop in their early teens. Financial capabilities could be built in schools to keep pace with today’s fast-changing and complex financial marketplace. Their financial decisions will influence their future economic well-being. This study examined the relative effectiveness of variations of a co-curricular financial literacy education program offered to eight graders of a Middle School in New England. Mixed methods were utilized first, to determine differences in program effects at improving the students’ financial literacy and changing their financial attitudes and behaviors; and second, to uncover the determinants of the outcomes in building the students’ financial capability. There were differences found in degrees of improvements in financial knowledge and financial attitudes between each one and another variant of the program. Intervening variables, including influences of the family and peers, having a job and access to money, were also found to affect the financial outcomes. (Author abstract)Bolanos, A.B. (2012). Key determinants in building financial capability among middle schoolers with a school-based financial literacy education program. Retrieved from http://academicarchive.snhu.eduDoctor of Philosophy (Ph.D.)School of Busines

Automated Analysis of Cryptococcal Macrophage Parasitism Using GFP-Tagged Cryptococci

The human fungal pathogens Cryptococcus neoformans and C. gattii cause life-threatening infections of the central nervous system. One of the major characteristics of cryptococcal disease is the ability of the pathogen to parasitise upon phagocytic immune effector cells, a phenomenon that correlates strongly with virulence in rodent models of infection. Despite the importance of phagocyte/Cryptococcus interactions to disease progression, current methods for assaying virulence in the acrophage system are both time consuming and low throughput. Here, we introduce the first stable and fully characterised GFP–expressing derivatives of two widely used cryptococcal strains: C. neoformans serotype A type strain H99 and C. gattii serotype B type strain R265. Both strains show unaltered responses to environmental and host stress conditions and no deficiency in virulence in the macrophage model system. In addition, we report the development of a method to effectively and rapidly investigate macrophage parasitism by flow cytometry, a technique that preserves the accuracy of current approaches but offers a four-fold improvement in speed

Therapeutic impact of cytoreductive surgery and irradiation of posterior fossa ependymoma in the molecular era: a retrospective multicohort analysis

PURPOSE: Posterior fossa ependymoma comprises two distinct molecular variants termed EPN_PFA and EPN_PFB that have a distinct biology and natural history. The therapeutic value of cytoreductive surgery and radiation therapy for posterior fossa ependymoma after accounting for molecular subgroup is not known. METHODS: Four independent nonoverlapping retrospective cohorts of posterior fossa ependymomas (n = 820) were profiled using genome-wide methylation arrays. Risk stratification models were designed based on known clinical and newly described molecular biomarkers identified by multivariable Cox proportional hazards analyses. RESULTS: Molecular subgroup is a powerful independent predictor of outcome even when accounting for age or treatment regimen. Incompletely resected EPN_PFA ependymomas have a dismal prognosis, with a 5-year progression-free survival ranging from 26.1% to 56.8% across all four cohorts. Although first-line (adjuvant) radiation is clearly beneficial for completely resected EPN_PFA, a substantial proportion of patients with EPN_PFB can be cured with surgery alone, and patients with relapsed EPN_PFB can often be treated successfully with delayed external-beam irradiation. CONCLUSION: The most impactful biomarker for posterior fossa ependymoma is molecular subgroup affiliation, independent of other demographic or treatment variables. However, both EPN_PFA and EPN_PFB still benefit from increased extent of resection, with the survival rates being particularly poor for subtotally resected EPN_PFA, even with adjuvant radiation therapy. Patients with EPN_PFB who undergo gross total resection are at lower risk for relapse and should be considered for inclusion in a randomized clinical trial of observation alone with radiation reserved for those who experience recurrence

Persistence on therapy and propensity matched outcome comparison of two subcutaneous interferon beta 1a dosages for multiple sclerosis

To compare treatment persistence between two dosages of interferon β-1a in a large observational multiple sclerosis registry and assess disease outcomes of first line MS treatment at these dosages using propensity scoring to adjust for baseline imbalance in disease characteristics. Treatment discontinuations were evaluated in all patients within the MSBase registry who commenced interferon β-1a SC thrice weekly (n = 4678). Furthermore, we assessed 2-year clinical outcomes in 1220 patients treated with interferon β-1a in either dosage (22 µg or 44 µg) as their first disease modifying agent, matched on propensity score calculated from pre-treatment demographic and clinical variables. A subgroup analysis was performed on 456 matched patients who also had baseline MRI variables recorded. Overall, 4054 treatment discontinuations were recorded in 3059 patients. The patients receiving the lower interferon dosage were more likely to discontinue treatment than those with the higher dosage (25% vs. 20% annual probability of discontinuation, respectively). This was seen in discontinuations with reasons recorded as “lack of efficacy” (3.3% vs. 1.7%), “scheduled stop” (2.2% vs. 1.3%) or without the reason recorded (16.7% vs. 13.3% annual discontinuation rate, 22 µg vs. 44 µg dosage, respectively). Propensity score was determined by treating centre and disability (score without MRI parameters) or centre, sex and number of contrast-enhancing lesions (score including MRI parameters). No differences in clinical outcomes at two years (relapse rate, time relapse-free and disability) were observed between the matched patients treated with either of the interferon dosages. Treatment discontinuations were more common in interferon β-1a 22 µg SC thrice weekly. However, 2-year clinical outcomes did not differ between patients receiving the different dosages, thus replicating in a registry dataset derived from “real-world” database the results of the pivotal randomised trial. Propensity score matching effectively minimised baseline covariate imbalance between two directly compared sub-populations from a large observational registry

Mutations in CENPE define a novel kinetochore-centromeric mechanism for microcephalic primordial dwarfism

Defects in centrosome, centrosomal-associated and spindle-associated proteins are the most frequent cause of primary microcephaly (PM) and microcephalic primordial dwarfism (MPD) syndromes in humans. Mitotic progression and segregation defects, microtubule spindle abnormalities and impaired DNA damage-induced G2-M cell cycle checkpoint proficiency have been documented in cell lines from these patients. This suggests that impaired mitotic entry, progression and exit strongly contribute to PM and MPD. Considering the vast protein networks involved in coordinating this cell cycle stage, the list of potential target genes that could underlie novel developmental disorders is large. One such complex network, with a direct microtubule-mediated physical connection to the centrosome, is the kinetochore. This centromeric-associated structure nucleates microtubule attachments onto mitotic chromosomes. Here, we described novel compound heterozygous variants in CENPE in two siblings who exhibit a profound MPD associated with developmental delay, simplified gyri and other isolated abnormalities. CENPE encodes centromere-associated protein E (CENP-E), a core kinetochore component functioning to mediate chromosome congression initially of misaligned chromosomes and in subsequent spindle microtubule capture during mitosis. Firstly, we present a comprehensive clinical description of these patients. Then, using patient cells we document abnormalities in spindle microtubule organization, mitotic progression and segregation, before modeling the cellular pathogenicity of these variants in an independent cell system. Our cellular analysis shows that a pathogenic defect in CENP-E, a kinetochore-core protein, largely phenocopies PCNT-mutated microcephalic osteodysplastic primordial dwarfism-type II patient cells. PCNT encodes a centrosome-associated protein. These results highlight a common underlying pathomechanism. Our findings provide the first evidence for a kinetochore-based route to MPD in humans

Punta morada de la papa, Marchitez letal de la palma aceitera y Síndrome de cuello virado de la papaya: enfermedades asociadas a fitoplasmas en Ecuador

Phytoplasma are wall-less bacteria limited to the phloem vessels in higher plants. Diseases associated with phytoplasma, in the past, have not been a serious problem in Ecuador. Nevertheless, for climate change effects, their importance has been increasing suddenly. This research was focused on the detection of phytoplasma using nested Polymerase Chain Reaction (PCR), in potato (Solanum tuberosum L.), oil palm (Elaeis guineensis), and papaya (Carica papaya L.) plants. Phytoplasma-specific PCR amplifications were generated by nested PCR in 50% samples of Twisted neck syndrome in papaya, 38% potato samples showing Purple top symptoms, and in all the samples showing symptoms of Lethal wilt of oil palm.  A continual increase in the incidence of Potato purple top was observed, and there is a high risk of contamination in the southern production zones of the country. In potato, results of this study are more closely related to Candidatus Phytoplasma subgroup 16SrI. Lethal wilt continues to be a major threat to oil palm production. In papaya, members of the group 16SrXIII-E produce a disease similar as reported in this study. Keywords: Candidatus Phytoplasma, crop production, nested PCRLos fitoplasmas son bacterias sin pared celular limitadas al floema en plantas superiores. Las enfermedades asociadas a fitoplasmas, en el pasado, han sido poco frecuentes y severas en el Ecuador. Sin embargo,  por efectos del cambio climático, su importancia se ha incrementado en un corto tiempo. Esta investigación tuvo el objetivo de detectar fitoplasmas mediante la reacción en cadena de la polimerasa (PCR) anidada, en plantas de  papa (Solanum tuberosum), palma aceitera (Elaeis guineensis) y papaya (Carica papaya). Amplicones del tamaño esperado para fitoplasma fueron obtenidos en el 50% de las muestras con síndrome de cuello virado de la papaya, 38% de plantas con sintomatología de punta morada de la papa y todas las muestras con marchitez letal de la palma aceitera. Punta morada ha ido incrementándose en incidencia y severidad, y existe alto riesgo de contaminación de la enfermedad en zonas de producción de semilla al centro-sur del país. En papa los resultados de este estudio están relacionados con Candidatus Phytoplasma sp. subgrupo 16SrI. La Marchitez letal continúa siendo una seria amenaza para el cultivo de palma aceitera. En papaya, miembros del 16SrXIII-E producen sintomatología similar a la informada en el presente trabajo. Palabras clave: Candidatus Phytoplasma, producción de plantas, PCR-anidad

Expression Levels of a Kinesin-13 Microtubule Depolymerase Modulates the Effectiveness of Anti-Microtubule Agents

Chemotherapeutic drugs often target the microtubule cytoskeleton as a means to disrupt cancer cell mitosis and proliferation. Anti-microtubule drugs inhibit microtubule dynamics, thereby triggering apoptosis when dividing cells activate the mitotic checkpoint. Microtubule dynamics are regulated by microtubule-associated proteins (MAPs); however, we lack a comprehensive understanding about how anti-microtubule agents functionally interact with MAPs. In this report, we test the hypothesis that the cellular levels of microtubule depolymerases, in this case kinesin-13 s, modulate the effectiveness of the microtubule disrupting drug colchicine.We used a combination of RNA interference (RNAi), high-throughput microscopy, and time-lapse video microscopy in Drosophila S2 cells to identify a specific MAP, kinesin-like protein 10A (KLP10A), that contributes to the efficacy of the anti-microtubule drug colchicine. KLP10A is an essential microtubule depolymerase throughout the cell cycle. We find that depletion of KLP10A in S2 cells confers resistance to colchicine-induced microtubule depolymerization to a much greater extent than depletion of several other destabilizing MAPs. Using image-based assays, we determined that control cells retained 58% (+/-2%SEM) of microtubule polymer when after treatment with 2 microM colchicine for 1 hour, while cells depleted of KLP10A by RNAi retained 74% (+/-1%SEM). Likewise, overexpression of KLP10A-GFP results in increased susceptibility to microtubule depolymerization by colchicine.Our results demonstrate that the efficacy of microtubule destabilization by a pharmacological agent is dependent upon the cellular expression of a microtubule depolymerase. These findings suggest that expression levels of Kif2A, the human kinesin-13 family member, may be an attractive biomarker to assess the effectiveness of anti-microtubule chemotherapies. Knowledge of how MAP expression levels affect the action of anti-microtubule drugs may prove useful for evaluating possible modes of cancer treatment