The molecular characterization of the glucoamylase and pyruvate kinase genes of Aspergillus niger

The glucoamylase gene of Aspergillus niger, glaA , is expressed at high levels in the presence of starch or maltose and repressed at the level of transcription by glucose. The promoter region of glaA was subjected to deletion and site-specific mutation analysis. Using autonomously replicating vectors with the A. niger lacA as the reporter gene, regulatory elements that control glaA expression in A. niger were mapped. Expression analysis showed that two elements that closely match the AnCF-binding consensus sequence were important for glaA expression. Site-directed mutagenesis of four elements that matched the CreA binding consensus showed that only two of these sites were important for regulation and that both sites were essential for glaA expression in maltose and glucose. Interestingly, one of the essential CreA sites overlapped the upstream portion of the single AmyR consensus element while the other site was immediately downstream from this site. The results of this study, therefore, suggested that both AnCF and AmyR were required for transcription of glaA and that regulation involved competition for overlapping binding sites between the transcriptional repressor CreA, and starch responsive transcription activator AmyR. The highly expressed pyruvate kinase gene of Aspergillus niger, pkiA , encodes an enzyme in glycolysis. This study found that pyruvate kinase expression was approximately 8-fold higher under glycolytic growth conditions, as compared to gluconeogenic growth conditions. Deletion analysis of the pkiA promoter region found three regions important for expression. One of the regions contains a 17 base pair sequence that is conserved in A. niger and A. nidulans . Another region contains a sequence closely matching the AnCF-binding consensu

Role of the Digital for the Third Sector in the Campania Region (Italy): From the Side of Research to the Side of the Objects in the Aftermath of the COVID-19Pandemic

This paper aims to investigate the role of the digital in facing the social and practical impact of the Covid-19 pandemic on particularly fragile and affected realities, Third Sector Organizations (TSO), in a context pervaded by structural and socio-economic weaknesses, the Campania Region in south Italy. It analyzes the digital object as: 1. a process - digitization/digitalization and digital transformation, 2. as a scenario - in which identities, activities, and alternative spaces of use or data collection transit and are constructed, and as 3. skills and capabilities - as social capital and value. This re-framing work is intended to be a useful tool for reflecting on the role of digital in research processes, while also offering a contribution to the debate on the Third Sector in contexts of crisis, such as the one generated by the COVID-19 pandemic, that impact its existence, activity, shape, and perspectives

Una propuesta para mejorar la situación del ingresante a la Universidad en la lectura de textos de matemática

Nuestro trabajo se centra en la utilización de algunos recursos de la Educación a Distancia para la Educación Presencial en pos del mejoramiento de la formación matemática de los egresados del nivel secundario. Desarrollamos un conjunto de materiales impresos, que también complementamos con recursos tecnológicos, lo suficientemente amigables para que quienes ingresarán próximamente a la educación superior mejoren su autoestima y motivación, comprendiendo cuestiones que creían inalcanzables. Describimos el contexto de trabajo, los conocimientos y formación de que disponen los egresados del nivel secundario, en cuanto a la matemática básica. El material es de acceso libre y gratuito en la Web y que constituye nuestro aporte concreto para ser utilizado en las escuelas secundarias o bien con los ingresantes a la Universidad en un curso introductorio de matemática. Concebimos un aula interactiva, con producción-elaboración conjunta del conocimiento, en la que el docente es mediador, democrático, donde el estudiante debe pasar de espectador pasivo a sujeto participativo. Esta ideología pedagógica debe revelarse con vivencias. Esta concepción sobre la enseñanza exige coherencia entre los pensamientos, las teorías y las prácticas que implementamos cotidianamente. Hemos realizado experiencias desde 2003. Consideramos que la propuesta constituye una herramienta más para trabajar las dificultades formativas con que los alumnos llegan a la Universidad, en cuanto al área que nos concierne que es una de las principales causas que conducen al abandono temprano de los estudios superiores. En cuanto al futuro, somos optimistas: con el paso de los años, la continuidad democrática y la educación obligatoria los jóvenes de nuestro país conseguirán esta formación imprescindible, no sólo para el acceso a niveles educativos más altos sino como derecho de todos los argentinos. Este año, tras la ayuda de la Secretaría de Políticas Universitarias de la Nación, hemos tenido la posibilidad de llevar esta iniciativa a 2.750 estudiantes de la última etapa del nivel medio trabajando junto a unos 130 profesores secundarios, en un total de 14 ciudades de la zona de influencia de nuestra Institución. Esperamos que la implementación de este proyecto se vea reflejada en la disminución de la tasa de abandono durante el primer año, para la cohorte 2019

Insights into the interaction of the N-terminal amyloidogenic polypeptide of ApoA-I with model cellular membranes

BACKGROUND: About twenty variants of apolipoprotein A-I (ApoA-I) are associated to hereditary systemic amyloidoses. Although the molecular bases of this disease are still largely unknown, it has been hypothesized that ApoA-I proteolysis is a key event in pathogenesis, since it triggers the release of an N-terminal fragment (80-100 residue long) that misfolds to form amyloid deposits in peripheral organs and tissues. It is also known that cell membrane lipids play a key role in the fibrillogenic pathway. In the case of ApoA-I related amyloidosis caused by L174S mutation, the 93-residue N-terminal fragment of ApoA-I ([1-93]ApoA-I) was found to be the major constituent of ex vivo fibrils. METHODS: With the main goal to investigate the interaction of either [1-93]ApoA-I and ApoA-I with biomimetic membranes, we set-up an experimental system based on the Raman Tweezers methodology. We tested GUVs composed by two types of zwitterionic lipids with a different fluidity degree, i.e. dioleoylphosphatidylcholine (DOPC) and dipalmitoylphosphatidylcholine (DPPC). RESULTS: We found that [1-93]ApoA-I induces conformational disorder in an ordered lipid bilayer. When interacting with fluid phases, instead, the fragment was found to be able to penetrate the membrane bilayer inducing an alignment of lipid chains. CONCLUSIONS: The interaction features of [1-93]ApoA-I with biomimetic membranes strongly depend on the lipid phase. Full-length ApoA-I was found to have similar effects, even if significantly less pronounced. GENERAL SIGNIFICANCE: Our observations shed light on still largely unknown molecular bases of ApoA-I fibrillogenic domain interaction with membranes

Celiac disease in type 1 diabetes mellitus

Celiac Disease (CD) occurs in patients with Type 1 Diabetes (T1D) ranging the prevalence of 4.4-11.1% versus 0.5% of the general population. The mechanism of association of these two diseases involves a shared genetic background: HLA genotype DR3-DQ2 and DR4-DQ8 are strongly associated with T1D, DR3-DQ2 with CD. The classical severe presentation of CD rarely occurs in T1D patients, but more often patients have few/mild symptoms of CD or are completely asymptomatic (silent CD). In fact diagnosis of CD is regularly performed by means of the screening in T1D patients. The effects of gluten-free diet (GFD) on the growth and T1D metabolic control in CD/T1D patient are controversial. Regarding of the GFD composition, there is a debate on the higher glycaemic index of gluten-free foods respect to gluten-containing foods; furthermore GFD could be poorer of fibers and richer of fat. The adherence to GFD by children with CD-T1D has been reported generally below 50%, lower respect to the 73% of CD patients, a lower compliance being more frequent among asymptomatic patients. The more severe problems of GFD adherence usually occur during adolescence when in GFD non compliant subjects the lowest quality of life is reported. A psychological and educational support should be provided for these patients

Doppler ultrasound venous mapping of the lower limbs

Aldo Innocente Galeandro1, Giovanni Quistelli2, Pietro Scicchitano2, Michele Gesualdo2, Annapaola Zito2, Paola Caputo2, Rosa Carbonara2, Giuseppe Galgano3, Francesco Ciciarello4, Sandro Mandolesi4, Claude Franceschi5, Marco Matteo Ciccone21Centro Diagnostica Globale and ASL-TA, Taranto, Italy; 2Cardiovascular Diseases Section, Department of Emergency and Organ Transplantation (DETO), University of Bari, Bari, Italy; 3Cardiovascular Diseases Section, Ente Ecclesiastico Ospedale Generale Regionale F Miulli, Acquaviva delle Fonti, Bari, Italy; 4Cardiology Department, Policlinico Umberto I, Sapienza University of Rome, Italy; 5Vascular Laboratories of Hospitals Saint Joseph and Pitié-Salpétrière, Paris, FranceBackground: The study aim was to test the accuracy (intra and interobserver variability), sensitivity, and specificity of a simplified noninvasive ultrasound methodology for mapping superficial and deep veins of the lower limbs.Methods: 62 consecutive patients, aged 62 ± 11 years, were enrolled. All underwent US-examinations, performed by two different investigators, of both legs, four anatomical parts, and 17 veins, to assess the interobserver variability of evaluation of superficial and deep veins of the lower limbs.Results: Overall the agreement between the second versus the first operator was very high in detecting reflux (sensitivity 97.9, specificity 99.7, accuracy 99.5; P = 0.80 at McNemar test). The higher CEAP classification stages were significantly associated with reflux (odds ratio: 1.778, 95% confidence interval: 1.552–2.038; P < 0.001) as well as with thrombosis (odds ratio: 2.765, 95% confidence interval: 1.741–4.389; P < 0.001). Thus, our findings show a strict association between the symptoms of venous disorders and ultrasound evaluation results for thrombosis or reflux.Conclusion: This study demonstrated that our venous mapping protocol is a reliable method showing a very low interobserver variability, which makes it accurate and reproducible for the assessment of the morphofunctional status of the lower limb veins.Keywords: venous mapping, new methodology, sensitivity, specificity, accurac

Microvascular Density, Endothelial Area, and Ki-67 Proliferative Index Correlate Each Other in Cat Post-Injection Fibrosarcoma

Soft tissue sarcomas are a large group of different tumor types both in humans and in animals. Among them, fibrosarcoma is the most frequent malignant mesenchymal tumoral form in cats, representing up to 28% of all cat skin tumors, while human fibrosarcoma, fortunately, only represents 5% of all sarcomas and 0.025% of the world-wide burden of tumors. This low incidence in humans leads to consideration of this group of tumoral diseases as rare, so therapeutic options are few due to the difficulty of starting clinical trials. In this context, the identification of research models for fibrosarcomas could be of great interest to deepen knowledge in this field and recognize new or possible biological pathways involved in tumor progression and metastasis. Angiogenesis is considered a fundamental scattering cause of tumor aggressiveness and progression in all forms of cancer, but only a few research parameters were developed and reported to express them quantitatively and qualitatively. The role in angiogenesis of microenvironmental stromal cells, such as fibroblasts, lymphocytes, mast cells, and macrophages, was largely demonstrated since this topic was first approached, while quantification of new vessels and their blood capacity in tumoral area is a relatively recent approach that could be well developed thanks to expertise in immunohistochemistry and image analysis. In this paper, a crossing study evaluating microvascular density (MVD), endothelial area (EA), and Ki-67 proliferative index was reported for a series of formalin-fixed and paraffin-embedded tissue samples from 99 cat patients, affected by cat post-injection fibrosarcoma, by using a till ×400 magnification light microscopy. We aim to demonstrate that cat pets may be considered a useful animal model for better studying the correspondent human diseases and we report, for the first time to our knowledge, experimental data in terms of correlation among MVD, EA, and Ki-67 strictly involved in aggressiveness and tumoral progression

A Long Baseline Neutrino Oscillation Experiment Using J-PARC Neutrino Beam and Hyper-Kamiokande

Document submitted to 18th J-PARC PAC meeting in May 2014. 50 pages, 41 figuresDocument submitted to 18th J-PARC PAC meeting in May 2014. 50 pages, 41 figuresDocument submitted to 18th J-PARC PAC meeting in May 2014. 50 pages, 41 figuresHyper-Kamiokande will be a next generation underground water Cherenkov detector with a total (fiducial) mass of 0.99 (0.56) million metric tons, approximately 20 (25) times larger than that of Super-Kamiokande. One of the main goals of Hyper-Kamiokande is the study of $CP$ asymmetry in the lepton sector using accelerator neutrino and anti-neutrino beams. In this document, the physics potential of a long baseline neutrino experiment using the Hyper-Kamiokande detector and a neutrino beam from the J-PARC proton synchrotron is presented. The analysis has been updated from the previous Letter of Intent [K. Abe et al., arXiv:1109.3262 [hep-ex]], based on the experience gained from the ongoing T2K experiment. With a total exposure of 7.5 MW $\times$ 10$^7$ sec integrated proton beam power (corresponding to $1.56\times10^{22}$ protons on target with a 30 GeV proton beam) to a $2.5$-degree off-axis neutrino beam produced by the J-PARC proton synchrotron, it is expected that the $CP$ phase $\delta_{CP}$ can be determined to better than 19 degrees for all possible values of $\delta_{CP}$, and $CP$ violation can be established with a statistical significance of more than $3\,\sigma$ ($5\,\sigma$) for $76$ ($58$) of the $\delta_{CP}$ parameter space

Measurement of νˉμ\bar{\nu}_{\mu} and νμ\nu_{\mu} charged current inclusive cross sections and their ratio with the T2K off-axis near detector

We report a measurement of cross section $\sigma(\nu_{\mu}+{\rm nucleus}\rightarrow\mu^{-}+X)$ and the first measurements of the cross section $\sigma(\bar{\nu}_{\mu}+{\rm nucleus}\rightarrow\mu^{+}+X)$ and their ratio $R(\frac{\sigma(\bar \nu)}{\sigma(\nu)})$ at (anti-)neutrino energies below 1.5 GeV. We determine the single momentum bin cross section measurements, averaged over the T2K $\bar{\nu}/\nu$-flux, for the detector target material (mainly Carbon, Oxygen, Hydrogen and Copper) with phase space restricted laboratory frame kinematics of $\theta_{\mu}$500 MeV/c. The results are $\sigma(\bar{\nu})=\left( 0.900\pm0.029{\rm (stat.)}\pm0.088{\rm (syst.)}\right)\times10^{-39}$ and $\sigma(\nu)=\left( 2.41\ \pm0.022{\rm{(stat.)}}\pm0.231{\rm (syst.)}\ \right)\times10^{-39}$in units of cm$^{2}$/nucleon and$R\left(\frac{\sigma(\bar{\nu})}{\sigma(\nu)}\right)= 0.373\pm0.012{\rm (stat.)}\pm0.015{\rm (syst.)}$.Comment: 18 pages, 8 figure

Measurements of neutrino oscillation in appearance and disappearance channels by the T2K experiment with 6.6 x 10(20) protons on target

111 pages, 45 figures, submitted to Physical Review D. Minor revisions to text following referee comments111 pages, 45 figures, submitted to Physical Review D. Minor revisions to text following referee comments111 pages, 45 figures, submitted to Physical Review D. Minor revisions to text following referee commentsWe thank the J-PARC staff for superb accelerator performance and the CERN NA61/SHINE Collaboration for providing valuable particle production data. We acknowledge the support of MEXT, Japan; NSERC, NRC, and CFI, Canada; CEA and CNRS/IN2P3, France; DFG, Germany; INFN, Italy; National Science Centre (NCN), Poland; RSF, RFBR and MES, Russia; MINECO and ERDF funds, Spain; SNSF and SER, Switzerland; STFC, UK; and the U. S. Deparment of Energy, USA. We also thank CERN for the UA1/NOMAD magnet, DESY for the HERA-B magnet mover system, NII for SINET4, the WestGrid and SciNet consortia in Compute Canada, GridPP, UK, and the Emerald High Performance Computing facility in the Centre for Innovation, UK. In addition, participation of individual researchers and institutions has been further supported by funds from ERC (FP7), EU; JSPS, Japan; Royal Society, UK; and DOE Early Career program, USA