Improving the PCR protocol to amplify a repetitive DNA sequence

Although PCR-based techniques have become an essential tool in the field of molecular and genetic research, the amplification of repetitive DNA sequences is limited. This is due to the truncated nature of the amplified sequences, which are also prone to errors during DNA polymerase-based amplification. The complex structure of repetitive DNA can form hairpin loops, which promote dissociation of the polymerase from the template, impairing complete amplification, and leading to the formation of incomplete fragments that serve as megaprimers. These megaprimers anneal with other sequences, generating unexpected fragments in each PCR cycle. Our gene model, MaSp1, is 1037-bp long, with 68% GC content, and its amino acid sequence is characterized by poly-alanine-glycine motifs, which represent the repetitive codon consensus. We describe the amplification of the MaSp1 gene through minor changes in the PCR program. The results show that a denaturation temperature of 98°C is the key determinant in the amplification of the MaSp1 partial gene sequence

Tumores óseos del pie

El manejo de los tumores óseos es difícil, debido a sus diferentes formas y comportamientos y a su baja incidencia. El pie no escapa a este axioma, ya que los tumores en esta región anatómica son particularmente infrecuentes y pueden mostrar un comportamiento clínico y radiológico diferente del que evidencian en otras regiones del esqueleto. Los autores revisan, en primer lugar, los aspectos generales de los tumores óseos del pie, considerando su epidemiología, el proceder diagnóstico y pronóstico y las opciones terapéuticas, para luego pasar a examinar las particularidades regionales e histológicas de estas neoplasias en relación con su tratamiento.The management of bone tumours is difficult, both because of their varying forms and behaviours and because of their low incidence. The foot is no exception to this axiom, as tumours in this particular anatomic region are particularly infrequent and may exhibit a clinical and radiological behaviour completely at variance to that in other skeletal regions. The authors first review the general aspects of the bobe tumours of the foot, considering their epidemiology, the diagnostic procedure and prognosis and the therapeutic options, and then examine the regional and histologic peculiaritiesof these neoplasms as related to their therapeutic management

Controls on intermontane basin filling, isolation and incision on the margin of the Puna Plateau, NW Argentina (similar to 23 degrees S)

Intermontane basins are illuminating stratigraphic archives of uplift, denudation and environmental conditions within the heart of actively growing mountain ranges. Commonly, however, it is difficult to determine from the sedimentary record of an individual basin whether basin formation, aggradation and dissection were controlled primarily by climatic, tectonic or lithological changes and whether these drivers were local or regional in nature. By comparing the onset of deposition, sediment-accumulation rates, incision, deformation, changes in fluvial connectivity and sediment provenance in two interrelated intermontane basins, we can identify diverse controls on basin evolution. Here, we focus on the Casa Grande basin and the adjacent Humahuaca basin along the eastern margin of the Puna Plateau in northwest Argentina. Underpinning this analysis is the robust temporal framework provided by U-Pb geochronology of multiple volcanic ashes and our new magnetostratigraphical record in the Humahuaca basin. Between 3.8 and 0.8 Ma, similar to 120 m of fluvial and lacustrine sediments accumulated in the Casa Grande basin as the rate of uplift of the Sierra Alta, the bounding range to its east, outpaced fluvial incision by the Rio Yacoraite, which presently flows eastward across the range into the Humahuaca basin. Detrital zircon provenance analysis indicates a progressive loss of fluvial connectivity from the Casa Grande basin to the downstream Humahuaca basin between 3 and 2.1 Ma, resulting in the isolation of the Casa Grande basin from 2.1 Ma to \u3c 1.7 Ma. This episode of basin isolation is attributed to aridification due to the uplift of the ranges to the east. Enhanced aridity decreased sediment supply to the Casa Grande basin to the point that aggradation could no longer keep pace with the rate of the surface uplift at the outlet of the basin. Synchronous events in the Casa Grande and Humahuaca basins suggest that both the initial onset of deposition above unconformities at similar to 3.8 Ma and the re-establishment of fluvial connectivity at similar to 0.8 Ma were controlled by climatic and/or tectonic changes affecting both basins. Reintegration of the fluvial network allowed subsequent incision in the Humahuaca basin to propagate upstream into the Casa Grande basin

Mood Disorders and Risk of Lung Cancer in the EAGLE Case-Control Study and in the U.S. Veterans Affairs Inpatient Cohort

Background: Mood disorders may affect lung cancer risk. We evaluated this hypothesis in two large studies. Methodology/Principal Findings: We examined 1,939 lung cancer cases and 2,102 controls from the Environment And Genetics in Lung cancer Etiology (EAGLE) case-control study conducted in Italy (2002-2005), and 82,945 inpatients with a lung cancer diagnosis and 3,586,299 person-years without a lung cancer diagnosis in the U.S. Veterans Affairs Inpatient Cohort (VA study), composed of veterans with a VA hospital admission (1969-1996). In EAGLE, we calculated odds ratios (ORs) and 95% confidence intervals (CI), with extensive adjustment for tobacco smoking and multiple lifestyle factors. In the VA study, we estimated lung cancer relative risks (RRs) and 95% CIs with time-dependent Poisson regression, adjusting for attained age, calendar year, hospital visits, time within the study, and related previous medical diagnoses. In EAGLE, we found decreased lung cancer risk in subjects with a personal history of mood disorders (OR: 0.59, 95% CI: 0.44-0.79, based on 121 lung cancer incident cases and 192 controls) and family history of mood disorders (OR: 0.62, 95% CI: 0.50-0.77, based on 223 lung cancer cases and 345 controls). The VA study analyses yielded similar results (RR: 0.74, 95% CI: 0.71-0.77, based on 2,304 incident lung cancer cases and 177,267 non-cancer person-years) in men with discharge diagnoses for mood disorders. History of mood disorders was associated with nicotine dependence, alcohol and substance use and psychometric scales of depressive and anxiety symptoms in controls for these studies. Conclusions/Significance: The consistent finding of a relationship between mood disorders and lung cancer risk across two large studies calls for further research into the complex interplay of risk factors associated with these two widespread and debilitating diseases. Although we adjusted for smoking effects in EAGLE, residual confounding of the results by smoking cannot be ruled out

Multiphase characterization of AGN winds in five local type-2 quasars

We present MEGARA integral field unit (IFU) observations of five local type-2 quasars (QSO2s, z ∼ 0.1) from the Quasar Feedback (QSOFEED) sample. These active galactic nuclei (AGN) have bolometric luminosities of 1045.5 − 46 erg s−1 and stellar masses of ∼1011 M⊙. The LR-V grating of MEGARA allows us to explore the kinematics of the ionized gas through the [O III]λ5007 Å emission line. The nuclear spectra of the five QSO2s, extracted in a circular aperture of ∼1.2″ (∼2.2 kpc) in diameter, matching the resolution of these seeing-limited observations, show signatures of high velocity winds in the form of broad (full width at half maximum, 1300 ≤ FWHM ≤ 2240 km s−1) and blueshifted components. We found that four out of the five QSO2s present outflows that we can resolve with our seeing-limited data, and they have radii ranging from 3.1 to 12.6 kpc. In the case of the two QSO2s with extended radio emission, we found that it is well aligned with the outflows, suggesting that low-power jets might be compressing and accelerating the ionized gas in these radio-quiet QSO2s. In the four QSO2s with spatially resolved outflows, we measured ionized mass outflow rates of 3.3–6.5 M⊙ yr−1 when we used [S II]-based densities, and of 0.7–1.6 M⊙ yr−1 when trans-auroral line-based densities were considered instead. We compared them with the corresponding molecular mass outflow rates (8–16 M⊙ yr−1), derived from CO(2–1) ALMA observations at 0.2″ resolution. The cold molecular outflows carry more mass than their ionized counterparts. However, both phases show lower outflow mass rates than those expected from observational scaling relations where uniform assumptions on the outflow properties were adopted. This might be indicating that the AGN luminosity is not the only driver of massive outflows and/or that these relations need to be rescaled using accurate outflow properties (i.e., electron density and radius). We did not find a significant impact of the outflows on the global star formation rates when considering the energy budget of the molecular and ionized outflows together. However, spatially resolved measurements of recent star formation in these targets are needed in order to evaluate this fairly, considering the dynamical timescales of the outflows, of 3–20 Myr for the ionized gas and 1–10 Myr for the molecular gas

Photoproduction of the f2(1270) resonance

We have performed a calculation of the γp→π+π−p reaction, where the two pions have been separated in D-wave producing the f2(1270) resonance. We use elements of the local hidden gauge approach that provides the interaction of vector mesons in which the f2(1270) resonance appears as a ρ-ρ molecular state in L=0 and spin 2. The vector meson dominance, incorporated in the local hidden gauge approach converts a photon into a ρ0 meson and the other meson connects the photon with the proton. The picture is simple and has no free parameters, since the parameters of the theory have been constrained in the previous study of the vector-vector states. In a second step we introduce new elements, not present in the local hidden gauge approach, adapting the ρ propagator to Regge phenomenology and introducing the ρNN tensor coupling. We find that both the differential cross section as well as the t dependence of the cross section are in good agreement with the experimental results and provide support for the molecular picture of the f2(1270) in the first baryonic reaction where it has been tested