High-Level Expression of Various Apolipoprotein (a) Isoforms by "Transferrinfection". The Role of Kringle IV Sequences in the Extracellular Association with Low-Density Lipoprotein

Characterization of the assembly of lipoprotein(a) [Lp(a)] is of fundamental importance to understanding the biosynthesis and metabolism of this atherogenic lipoprotein. Since no established cell lines exist that express Lp(a) or apolipoprotein(a) [apo(a)], a "transferrinfection" system for apo(a) was developed utilizing adenovirus receptor- and transferrin receptor-mediated DNA uptake into cells. Using this method, different apo(a) cDNA constructions of variable length, due to the presence of 3, 5, 7, 9, 15, or 18 internal kringle IV sequences, were expressed in cos-7 cells or CHO cells. All constructions contained kringle IV-36, which includes the only unpaired cysteine residue (Cys-4057) in apo(a). r-Apo(a) was synthesized as a precursor and secreted as mature apolipoprotein into the medium. When medium containing r-apo(a) with 9, 15, or 18 kringle IV repeats was mixed with normal human plasma LDL, stable complexes formed that had a bouyant density typical of Lp(a). Association was substantially decreased if Cys-4057 on r-apo(a) was replaced by Arg by site-directed mutagenesis or if Cys-4057 was chemically modified. Lack of association was also observed with r-apo(a) containing only 3, 5, or 7 kringle IV repeats without "unique kringle IV sequences", although Cys-4057 was present in all of these constructions. Synthesis and secretion of r-apo(a) was not dependent on its sialic acid content. r-Apo(a) was expressed even more efficiently in sialylation-defective CHO cells than in wild-type CHO cells. In transfected CHO cells defective in the addition of N-acetylglucosamine, apo(a) secretion was found to be decreased by 50%. Extracellular association with LDL was not affected by the carbohydrate moiety of r-apo(a), indicating a protein-protein interaction between r-apo(a) and apoB. These results show that, besides kringle IV-36, other kringle IV sequences are necessary for the extracellular association of r-apo(a) with LDL. Changes in the carbohydrate moiety of apo(a), however, do not affect complex formation

The Search for Stable, Massive, Elementary Particles

In this paper we review the experimental and observational searches for stable, massive, elementary particles other than the electron and proton. The particles may be neutral, may have unit charge or may have fractional charge. They may interact through the strong, electromagnetic, weak or gravitational forces or through some unknown force. The purpose of this review is to provide a guide for future searches - what is known, what is not known, and what appear to be the most fruitful areas for new searches. A variety of experimental and observational methods such as accelerator experiments, cosmic ray studies, searches for exotic particles in bulk matter and searches using astrophysical observations is included in this review.Comment: 34 pages, 8 eps figure

Loss of Num1-mediated cortical dynein anchoring negatively impacts respiratory growth

Num1 is a multifunctional protein that both tethers mitochondria to the plasma membrane and anchors dynein to the cell cortex during nuclear inheritance. Previous work has examined the impact loss of Num1-based mitochondrial tethering has on dynein function in Saccharomyces cerevisiae; here, we elucidate its impact on mitochondrial function. We find that like mitochondria, Num1 is regulated by changes in metabolic state, with the protein levels and cortical distribution of Num1 differing between fermentative and respiratory growth conditions. In cells lacking Num1, we observe a reproducible respiratory growth defect, suggesting a role for Num1 in not only maintaining mitochondrial morphology, but also function. A structure–function approach revealed that, unexpectedly, Num1- mediated cortical dynein anchoring is important for normal growth under respiratory conditions. The severe respiratory growth defect in Δnum1 cells is not specifically due to the canonical functions of dynein in nuclear migration but is dependent on the presence of dynein, as deletion of DYN1 in Δnum1 cells partially rescues respiratory growth. We hypothesize that misregulated dynein present in cells that lack Num1 negatively impacts mitochondrial function resulting in defects in respiratory growth

Evaluation of a novel mitochondrial Pan-Mucorales marker for the detection, identification, quantification, and growth stage determination of mucormycetes

Mucormycosis infections are infrequent yet aggressive and serious fungal infections. Early diagnosis of mucormycosis and its discrimination from other fungal infections is required for targeted treatment and more favorable patient outcomes. The majority of the molecular assays use 18 S rDNA. In the current study, we aimed to explore the potential of the mitochondrial rnl (encoding for large-subunit-ribosomal-RNA) gene as a novel molecular marker suitable for research and diagnostics. Rnl was evaluated as a marker for: (1) the Mucorales family, (2) species identification (Rhizopus arrhizus, R. microsporus, Mucor circinelloides, and Lichtheimia species complexes), (3) growth stage, and (4) quantification. Sensitivity, specificity, discriminatory power, the limit of detection (LoD), and cross-reactivity were evaluated. Assays were tested using pure cultures, spiked clinical samples, murine organs, and human paraffin-embedded-tissue (FFPE) samples. Mitochondrial markers were found to be superior to nuclear markers for degraded samples. Rnl outperformed the UMD universal® (Molyzm) marker in FFPE (71.5% positive samples versus 50%). Spiked blood samples highlighted the potential of rnl as a pan-Mucorales screening test. Fungal burden was reproducibly quantified in murine organs using standard curves. Identification of pure cultures gave a perfect (100%) correlation with the detected internal transcribed spacer (ITS) sequence. In conclusion, mitochondrial genes, such as rnl, provide an alternative to the nuclear 18 S rDNA genes and deserve further evaluation.CD laboratory: This research was funded by the Christian Doppler Laboratory for fungal infections

Differences in Hemodynamic, Hormonal and Heart Rate Variability Parameters in Complication-Free Pregnancies Compared to Individuals with Gestational Diabetes Mellitus and Preeclampsia : An Observational Retrospective Analysis

To investigate differences in hemodynamic, hormonal and heart rate variability parameters in women following complication-free pregnancies (healthy), preeclampsia and gestational diabetes mellitus (GDM) after giving childbirth. Data of 60 women (healthy: n = 29, age 32.7 ± 4.5 years, BMI 24.2 ± 4.3 kg/m(2); preeclampsia: n = 16, age 35.3 ± 4.4 years, 28.5 ± 6.4 kg/m(2); GDM, n = 15, age 32.3 ± 6.0 years, BMI 26.4 ± 6.2 kg/m(2)) were included. Two visits were conducted 16 and 48 weeks after giving childbirth. Hair samples were taken for analysis of cortisol and testosterone. ECG and blood pressure were recorded at each visit. Data were analyzed via RM-ANOVA and post-hoc testing (p ≤ 0.05). Heart rate increased from visit 1 to visit 2, whereas SDNN decreased (both p = 0.03). RMSSD showed an increased trend for groups (p = 0.06). Testosterone in the GDM group was significantly higher compared to the other groups (p = 0.002). Cortisol levels were significantly higher following post-hoc testing GDM was different compared to healthy individuals (p = 0.02). Hemodynamic changes from week 16 to week 48 did not differ between groups (p > 0.05). No differences between individuals with preeclampsia and healthy individuals were found for all hemodynamic parameters (p > 0.05). The study showed higher levels of chronic stress indicators in GDM measured via heart rate variability and cortisol compared to women with a history of preeclampsia and healthy women

Power Test of the First Two HL-LHC Insertion Quadrupole Magnets Built at CERN

The High-Luminosity project (HL-LHC) of the CERN Large Hadron Collider (LHC), requires low β* quadrupole magnets in Nb$_3$Sn technology that will be installed on each side of the ATLAS and CMS experiments. After a successful shortmodel magnet manufacture and test campaign, the project has advanced with the production, assembly, and test of full-size 7.15- m-long magnets. In the last two years, two CERN-built prototypes (MQXFBP1 and MQXFBP2) have been tested and magnetically measured at the CERN SM18 test facility. These are the longest accelerator magnets based on Nb$_3$Sn technology built and tested to date. In this paper, we present the test and analysis results of these two magnets, with emphasis on quenches and training, voltage-current measurements and the quench localization with voltage taps and a new quench antenna

Increased neutrophil-lymphocyte ratio is a poor prognostic factor in patients with primary operable and inoperable pancreatic cancer

Background: The neutrophil-lymphocyte ratio (NLR) has been proposed as an indicator of systemic inflammatory response. Previous findings from small-scale studies revealed conflicting results about its independent prognostic significance with regard to different clinical end points in pancreatic cancer (PC) patients. Therefore, the aim of our study was the external validation of the prognostic significance of NLR in a large cohort of PC patients. Methods: Data from 371 consecutive PC patients, treated between 2004 and 2010 at a single centre, were evaluated retrospectively. The whole cohort was stratified into two groups according to the treatment modality. Group 1 comprised 261 patients with inoperable PC at diagnosis and group 2 comprised 110 patients with surgically resected PC. Cancer-specific survival (CSS) was assessed using the Kaplan–Meier method. To evaluate the independent prognostic significance of the NLR, the modified Glasgow prognostic score (mGPS) and the platelet-lymphocyte ratio univariate and multivariate Cox regression models were applied. Results: Multivariate analysis identified increased NLR as an independent prognostic factor for inoperable PC patients (hazard ratio (HR)=2.53, confidence interval (CI)=1.64–3.91, P<0.001) and surgically resected PC patients (HR=1.61, CI=1.02–2.53, P=0.039). In inoperable PC patients, the mGPS was associated with poor CSS only in univariate analysis (HR=1.44, CI=1.04–1.98). Conclusion: Risk prediction for cancer-related end points using NLR does add independent prognostic information to other well-established prognostic factors in patients with PC, regardless of the undergoing therapeutic modality. Thus, the NLR should be considered for future individual risk assessment in patients with PC

Study of the Stabilization to the Nanometer Level of Mechanical Vibrations of the CLIC Main Beam

Original publication available at http://www.jacow.org/International audienceTo reach the design luminosity of CLIC, the movements of the quadrupoles should be limited to the nanometre level in order to limit the beam size and emittance growth. Below 1 Hz, the movements of the main beam quadrupoles will be corrected by a beambased feedback. But above 1 Hz, the quadrupoles should be mechanically stabilized. A collaboration effort is ongoing between several institutes to study the feasibility of the "nanostabilization" of the CLIC quadrupoles. The study described in this paper covers the characterization of independent measuring techniques including optical methods to detect nanometre sized displacements and analyze the vibrations. Actuators and feedback algorithms for sub-nanometre movements of magnets with a mass of more than 400 kg are being developed and tested. Input is given to the design of the quadrupole magnets, the supports and alignment system in order to limit the amplification of the vibration sources at resonant frequencies. A full scale mock-up integrating all these features is presently under design. Finally, a series of experiments in accelerator environments should demonstrate the feasibility of the nanometre stabilization

Coherent pion production in neutrino nucleus collision in the 1 GeV region

We calculate cross sections for coherent pion production in nuclei induced by neutrinos and antineutrinos of the electron and muon type. The analogies and differences between this process and the related ones of coherent pion production induced by photons, or the (p,n) and $(^3 He, t)$ reactions are discussed. The process is one of the several ones occurring for intermediate energy neutrinos, to be considered when detecting atmospheric neutrinos. For this purpose the results shown here can be easily extrapolated to other energies and other nuclei.Comment: 13 pages, LaTex, 8 post-script figures available at [email protected]