Highly specific PCR-RFLP assays for karyotyping the widespread 2Rb inversion in malaria vectors of the Anopheles gambiae complex

Background: Chromosomal inversion polymorphisms play a role in adaptation to heterogeneous environments. Inversion polymorphisms are implicated in the very high ecological flexibility of the three main malaria vector species of the Afrotropical Anopheles gambiae complex, facilitating the exploitation of anthropogenic environmental modifications and promoting a strong association with humans. In addition to extending the species' spatial and temporal distribution, inversions are associated with epidemiologically relevant mosquito behavior and physiology, underscoring their medical importance. We here present novel PCR-RFLP based assays strongly predictive of genotype for the cosmopolitan 2Rb inversion in An. coluzzii and An. gambiae, a development which overcomes the numerous constraints inherent to traditional cytological karyotyping. Methods: We designed PCR-RFLP genotyping assays based on tag SNPs previously computationally identified as strongly predictive (> 95%) of 2Rb genotype. We targeted those tags whose alternative allelic states destroyed or created the recognition site of a commercially available restriction enzyme, and designed assays with distinctive cleavage profiles for each inversion genotype. The assays were validated on 251 An. coluzzii and 451 An. gambiae cytologically karyotyped specimens from nine countries across Africa and one An. coluzzii laboratory colony. Results: For three tag SNPs, PCR-RFLP assays (denoted DraIII, MspAI, and TatI) reliably produced robust amplicons and clearly distinguishable electrophoretic profiles for all three inversion genotypes. Results obtained with the DraIII assay are ≥ 95% concordant with cytogenetic assignments in both species, while MspAI and TatI assays produce patterns highly concordant with cytogenetic assignments only in An. coluzzii or An. gambiae, respectively. Joint application of species-appropriate pairs of assays increased the concordance levels to > 99% in An. coluzzii and 98% in An. gambiae. Potential sources of discordance (e.g. imperfect association between tag and inversion, allelic dropout, additional polymorphisms in the restriction target site, incomplete or failed restriction digestion) are discussed. Conclusions: The availability of highly specific, cost effective and accessible molecular assays for genotyping 2Rb in An. gambiae and An. coluzzii allows karyotyping of both sexes and all developmental stages. These novel tools will accelerate deeper investigations into the role of this ecologically and epidemiologically important chromosomal inversion in vector biology.[Figure not available: see fulltext.

Histopathological features of gastrointestinal mucosal biopsies in children with juvenile idiopathic arthritis

BACKGROUND: The association between inflammatory bowel disease and joint involvement is well established. There is a paucity of data describing histopathological features of the gut in relation to juvenile idiopathic arthritis (JIA). METHODS: We retrospectively identified 33 (21 male) children aged 3-16 y with JIA (11 with oligoarthritis, 5 with polyarthritis, 8 with systemic onset arthritis, 8 with enthesitis-related arthritis (ERA), and 1 with psoriatic arthritis) with significant gastrointestinal (GI) symptoms who underwent upper and/or lower endoscopy. The histopathological findings were reviewed in addition to presence of autoantibodies and concomitant treatment. RESULTS: The most common GI indications for endoscopy were persistent abdominal pain (14/33 (42%)) and diarrhea (10/33 (30%)). Of the 33 children, 28 (85%) had gut mucosal inflammation, mostly affecting the colon (80%). Active inflammation of the gut was found in 5 of 28 (17%) children, and 15 of 28 (53%) children showed mild nonspecific inflammation. Eight patients (27%) had predominantly an eosinophilic infiltrate. Twenty-six patients had previously received treatment for JIA. There was a negative association with the use of immunomodulators and the presence of eosinophil inflammation. CONCLUSION: The majority of children with JIA and GI symptoms have histological evidence of mild nonspecific inflammation, but some having active colitis and prominent eosinophil infiltrate

Charge transfer and Fermi level shift in p-doped single-walled carbon nanotubes

The electronic properties of p-doped single-walled carbon nanotube (SWNT) bulk samples were studied by temperature-dependent resistivity and thermopower, optical reflectivity, and Raman spectroscopy. These all give consistent results for the Fermi level downshift (Delta E(F)) induced by doping. We find Delta E(F) approximate to 0.35 eV and 0.50 eV for concentrated nitric and sulfuric acid doping respectively. With these values, the evolution of Raman spectra can be explained by variations in the resonance condition as E(F) moves down into the valence band. Furthermore, we find no evidence for diameter-selective doping, nor any distinction between doping responses of metallic and semiconducting tubes

Fixed-parameter tractability of multicut parameterized by the size of the cutset

Given an undirected graph $G$, a collection $\{(s_1,t_1),..., (s_k,t_k)\}$ of pairs of vertices, and an integer $p$, the Edge Multicut problem ask if there is a set $S$ of at most $p$ edges such that the removal of $S$ disconnects every $s_i$ from the corresponding $t_i$. Vertex Multicut is the analogous problem where $S$ is a set of at most $p$ vertices. Our main result is that both problems can be solved in time $2^{O(p^3)}... n^{O(1)}$, i.e., fixed-parameter tractable parameterized by the size $p$ of the cutset in the solution. By contrast, it is unlikely that an algorithm with running time of the form $f(p)... n^{O(1)}$ exists for the directed version of the problem, as we show it to be W[1]-hard parameterized by the size of the cutset

Dynamics of Bianchi type I elastic spacetimes

We study the global dynamical behavior of spatially homogeneous solutions of the Einstein equations in Bianchi type I symmetry, where we use non-tilted elastic matter as an anisotropic matter model that naturally generalizes perfect fluids. Based on our dynamical systems formulation of the equations we are able to prove that (i) toward the future all solutions isotropize; (ii) toward the initial singularity all solutions display oscillatory behavior; solutions do not converge to Kasner solutions but oscillate between different Kasner states. This behavior is associated with energy condition violation as the singularity is approached.Comment: 28 pages, 11 figure

Optical properties of the vibrations in charged C60_{60} molecules

The transition strengths for the four infrared-active vibrations of charged C$_{60}$ molecules are evaluated in self-consistent density functional theory using the local density approximation. The oscillator strengths for the second and fourth modes are strongly enhanced relative to the neutral C$_{60}$ molecule, in good agreement with the experimental observation of ``giant resonances'' for those two modes. Previous theory, based on a ``charged phonon'' model, predicted a quadratic dependence of the oscillator strength on doping, but this is not borne out in our calculations.Comment: 10 pages, RevTeX3.

Fatigue strengthening of damaged steel members using wire arc additive manufacturing

In this study, a directed energy deposition (DED) process called wire arc additive manufacturing (WAAM) is employed for the fatigue strengthening of damaged steel members. Three steel specimens with central cracks were tested under a high-cycle fatigue loading (HCF) regime: (1) the reference specimen; (2) the WAAM-repaired specimen with an as-deposited profile, and (3) the WAAM-repaired specimen machined to reduce stress concentration factors (SCF). The corresponding finite element (FE) simulation of the WAAM process was calibrated using static experimental results, which revealed the main mechanism. The process was found to introduce compressive residual stresses at the crack tip owing to the thermal contraction of the repair. The FE results also revealed that stress concentration exists at the root of the as-deposited WAAM; this stress concentration can be mitigated by machining the WAAM to a pyramid-like shape. The fractography analysis indicated that the cracks were initiated at the WAAM-steel interface, and microscopic observations revealed that the microcracks were arrested by the porosities in the melted interface. The results of this pioneering study suggest that WAAM repair is a promising technique for combating fatigue damage in steel structures

Feasibilty of Transcutaneous pCO2 Monitoring During Immediate Transition After Birth\u2014A Prospective Observational Study

Background: According to recommendations, non-invasive monitoring during neonatal resuscitation after birth includes heart rate (HR) and oxygen saturation (SpO2). Continuous transcutaneous monitoring of carbon dioxide partial pressure (tcpCO2) may further offer quantitative information on neonatal respiratory status. Objective: We aimed to investigate feasibility of tcpCO2 measurements in the delivery room during immediate neonatal transition and to compare the course of tcpCO2 between stable term and preterm infants. Methods: Neonates without need for cardio-respiratory intervention during immediate transition after birth were enrolled in a prospective observational study. In these term and preterm neonates, we measured HR and SpO2 by pulse oximetry on the right wrist and tcpCO2 with the sensor applied on the left hemithorax during the first 15 min after birth. Courses of tcpCO2 were analyzed in term and preterm neonates and groups were compared. Results: Fifty-three term (gestational age: 38.8 \ub1 0.9 weeks) and 13 preterm neonates (gestational age: 34.1 \ub1 1.5 weeks) were included. First tcpCO2 values were achieved in both groups at minute 4 after birth, which reached a stable plateau after the equilibration phase at minute 9. Mean tcpCO2 values 15 min after birth were 46.2 (95% CI 34.5\u201357.8) mmHg in term neonates and 48.5 (95%CI 43.0\u201354.1) mmHg in preterm neonates. Preterm and term infants did not show significant differences in the tcpCO2 values at any time point. Conclusion: This study demonstrates that tcpCO2 measurement is feasible during immediate neonatal transition after birth and that tcpCO2 values were comparable in stable term and preterm neonates