Shearographic non-destructive evaluation of space shuttle thermal protection systems

Preliminary results of shearographic inspections of the shuttle external tank (ET) spray-on foam insulation (SOFI) and solid rocket booster (SRB) Marshall sprayable ablative (MSA-2) epoxy-cork thermal protection systems (TPS) are presented. Debonding SOFI or MSA-2 damage the orbiter 'belly' tile and exposes the ET/SRB to thermal loading. Previous work with the ET/SRB showed promising results with shearography. The first area investigated was the jack pad close-out, one of many areas on the ET where foam is applied at KSC. Voids 0.375 inch were detected in 1.75 inch thick foam using a pressure reduction of less than 0.4 psi. Of primary interest are areas of the ET that directly face the orbiter tile TPS. It is estimated that 90% of tile TPS damage on the orbiter 'belly' results from debonding SOFI during ascent. Test panels modeling these areas were manufactured with programmed debonds to determine the sensitivity of shearography as a function of debond size, SOFI thickness and vacuum. Results show repeatable detection of debonds with a diameter approximately half the SOFI thickness at less than 0.4 psi pressure reduction. Preliminary results are also presented on inspections of MSA-2 and the remote manipulator system (RMS) honeycomb materia

Hydrolysis of tannic acid catalyzed by immobilized-stabilized derivatives of tannase from Lactobacillus plantarum

A recombinant tannase from Lactobacillus plantarum, overexpressed in Escherichia coli, was purified in a single step by metal chelate affinity chromatography on poorly activated nickel supports. It was possible to obtain 0.9 g of a pure enzyme by using only 20 mL of chromatographic support. The pure enzyme was immobilized and stabilized by multipoint covalent immobilization on highly activated glyoxyl agarose. Derivatives obtained by multipoint and multisubunit immobilization were 500- and 1000-fold more stable than both the soluble enzyme and the one-point-immobilized enzyme in experiments of thermal and cosolvent inactivation, respectively. In addition, up to 70 mg of pure enzyme was immobilized on 1 g of wet support. The hydrolysis of tannic acid was optimized by using the new immobilized tannase derivative. The optimal reaction conditions were 30% diglyme at pH 5.0 and 4 C. Under these conditions, it was possible to obtain 47.5 mM gallic acid from 5 mM tannic acid as substrate. The product was pure as proved by HPLC. On the other hand, the immobilized biocatalyst preserved >95% of its initial activity after 1 month of incubation under the optimal reaction conditionsThis work was supported by Grants AGL2008-01052, AGL-2009-07625, Consolider INGENIO 2010 CSD2007-00063 FUN-C-FOOD(CICYT),RM2008-00002 (INIA), and S-0505/AGR/000153 (CAM). J.A.C. is the recipient of a predoctoral fellowship from the I3P-CSIC Program and FPI-MEC, and G.F.-L. and L.B. are recipients of Ramon y Cajal postdoctoral contracts.Peer reviewe

Intrastructural help: harnessing T helper cells induced by licensed vaccines for improvement of HIV env antibody responses to virus-like particle vaccines

Induction of persistent antibody responses by vaccination is generally thought to depend on efficient help by T follicular helper cells. Since the T helper cell response to HIV Env may not be optimal, we explored the possibility of improving the HIV Env antibody response to virus-like particle (VLP) vaccines by recruiting T helper cells induced by commonly used licensed vaccines to provide help for Env-specific B cells. B cells specific for the surface protein of a VLP can internalize the entire VLP and thus present peptides derived from the surface and core proteins on their major histocompatibility complex class II (MHC-II) molecules. This allows T helper cells specific for the core protein to provide intrastructural help for B cells recognizing the surface protein. Consistently, priming mice with an adjuvanted Gag protein vaccine enhanced the HIV Env antibody response to subsequent booster immunizations with HIV VLPs. To harness T helper cells induced by the licensed Tetanolpur vaccines, HIV VLPs that contained T helper cell epitopes of tetanus toxoid were generated. Tetanol-immunized mice raised stronger antibody responses to immunizations with VLPs containing tetanus toxoid T helper cell epitopes but not to VLPs lacking these epitopes. Depending on the priming immunization, the IgG subtype response to HIV Env after the VLP immunization could also be modified. Thus, harnessing T helper cells induced by other vaccines appears to be a promising approach to improve the HIV Env antibody response to VLP vaccines

Targeting Antibody Responses to the Membrane Proximal External Region of the Envelope Glycoprotein of Human Immunodeficiency Virus

Although human immunodeficiency type 1 (HIV-1) infection induces strong antibody responses to the viral envelope glycoprotein (Env) only a few of these antibodies possess the capacity to neutralize a broad range of strains. The induction of such antibodies represents an important goal in the development of a preventive vaccine against the infection. Among the broadly neutralizing monoclonal antibodies discovered so far, three (2F5, Z13 and 4E10) target the short and hidden membrane proximal external region (MPER) of the gp41 transmembrane protein. Antibody responses to MPER are rarely observed in HIV-infected individuals or after immunization with Env immunogens. To initiate antibody responses to MPER in its membrane-embedded native conformation, we generated expression plasmids encoding the membrane-anchored ectodomain of gp41 with N-terminal deletions of various sizes. Following transfection of these plasmids, the MPER domains are displayed on the cell surface and incorporated into HIV virus like particles (VLP). Transfected cells displaying MPER mutants bound as efficiently to both 2F5 and 4E10 as cells transfected with a plasmid encoding full-length Env. Mice immunized with VLPs containing the MPER mutants produced MPER-specific antibodies, the levels of which could be increased by the trimerization of the displayed proteins as well as by a DNA prime-VLP boost immunization strategy. Although 2F5 competed for binding to MPER with antibodies in sera of some of the immunized mice, neutralizing activity could not be detected. Whether this is due to inefficient binding of the induced antibodies to MPER in the context of wild type Env or whether the overall MPER-specific antibody response induced by the MPER display mutants is too low to reveal neutralizing activity, remains to be determined

A barcoded flow cytometric assay to explore the antibody responses against SARS-CoV-2 spike and its variants

The SARS-CoV-2 pandemic has spread to all parts of the world and can cause life-threatening pneumonia and other severe disease manifestations known as COVID-19. This health crisis has resulted in a significant effort to stop the spread of this new coronavirus. However, while propagating itself in the human population, the virus accumulates mutations and generates new variants with increased fitness and the ability to escape the human immune response. Here we describe a color-based barcoded spike flow cytometric assay (BSFA) that is particularly useful to evaluate and directly compare the humoral immune response directed against either wild type (WT) or mutant spike (S) proteins or the receptor-binding domains (RBD) of SARS-CoV-2. This assay employs the human B lymphoma cell line Ramos, transfected for stable expression of WT or mutant S proteins or a chimeric RBD-CD8 fusion protein. We find that the alpha and beta mutants are more stably expressed than the WT S protein on the Ramos B cell surface and/or bind with higher affinity to the viral entry receptor ACE2. However, we find a reduce expression of the chimeric RBD-CD8 carrying the point mutation N501Y and E484K characteristic for the alpha and beta variant, respectively. The comparison of the humoral immune response of 12 vaccinated probands with 12 COVID-19 patients shows that after the boost, the S-specific IgG class immune response in the vaccinated group is similar to that of the patient group. However, in comparison to WT the specific IgG serum antibodies bind less well to the alpha variant and only poorly to the beta variant S protein. This is in line with the notion that the beta variant is an immune escape variant of SARS-CoV-2. The IgA class immune response was more variable than the IgG response and higher in the COVID-19 patients than in the vaccinated group. In summary, we think that our BSFA represents a useful tool to evaluate the humoral immunity against emerging variants of SARS-CoV-2 and to analyze new vaccination protocols against these variants

Биогеохимические исследования урбанизированных ландшафтов восточной Финляндии

Объектом исследования являются урбанизированные ландшафты трех городов Восточной Финляндии г. Юука, г. Оутокумпу, г. Миккели. Цель работы - биогеохимические исследования урбанизированных ландшафтов восточной Финляндии по результатам исследования проб почв, листьев и стеблей черники, лишайника, листьев древесных растений. В процессе исследования проводились обзор и анализ ранее проведенных работ, анализы литературных, нормативных и фондовых источников, анализировались статистических данные и элементный состав комплекса природных сред. В результате исследования была изучена геоэкологическая обстановка урбанизированных территорий Восточной Финляндии Степень внедрения результатов: полученный анализ эколого-геохимической ситуации урбанизированных территорий Финляндии является составной частью комплексного эколого-геохимического мониторинга территорий, проводимого кафедрой ГЭГХ ТПУ. Аналитический материал будет использован в учебном процессе. Область применения: результаты эколого-геохимического мониторинга урбанизированных территорий Финляндии могут быть использованы природоохранными организациями для ориентировочных данных по содержанию химических элементов в почвах и живых объектах городских агломераций.Object of research are urban landscapes of the three cities in the Eastern Finland city of Juuka, in Outokumpu, the city of Mikkeli. Purpose - biogeochemical studies of the urban landscape of Eastern Finland the results of the research of samples of soil, leaves and stems of bilberry, lichen, leaves of woody plants. In the process of investigation the review and analysis of earlier conducted works, literary analyses, regulatory and archival sources, analyzed statistical data and elemental composition of complex natural environments. The study was studied geo-ecological situation in the urbanized territories of Eastern Finland level of implementation of results: the analysis of environmental and geochemical situation in the urbanized territories of Finland is part of an integrated ecological-geochemical monitoring of the territories held by the Department GEGH TPU. Analytical material will be used in the educational process. Application field: the results of ecological-geochemical monitoring in the urbanized territories of Finland can be used by conservation organizations for estimates on the content of chemical elements in the soils and the living facilities of urban agglomerations

Search for antihelium in cosmic rays

The Alpha Magnetic Spectrometer (AMS) was flown on the space shuttle Discovery during flight STS-91 in a 51.7 degree orbit at altitudes between 320 and 390 km. A total of 2.86 * 10^6 helium nuclei were observed in the rigidity range 1 to 140 GV. No antihelium nuclei were detected at any rigidity. An upper limit on the flux ratio of antihelium to helium of < 1.1 * 10^-6 is obtained.Comment: 18 pages, Latex, 9 .eps figure

Structure-based programming of lymph-node targeting in molecular vaccines

In cancer patients, visual identification of sentinel lymph nodes (LNs) is achieved by the injection of dyes that bind avidly to endogenous albumin, targeting these compounds to LNs, where they are efficiently filtered by resident phagocytes1, 2. Here we translate this ‘albumin hitchhiking’ approach to molecular vaccines, through the synthesis of amphiphiles (amph-vaccines) comprising an antigen or adjuvant cargo linked to a lipophilic albumin-binding tail by a solubility-promoting polar polymer chain. Administration of structurally optimized CpG-DNA/peptide amph-vaccines in mice resulted in marked increases in LN accumulation and decreased systemic dissemination relative to their parent compounds, leading to 30-fold increases in T-cell priming and enhanced anti-tumour efficacy while greatly reducing systemic toxicity. Amph-vaccines provide a simple, broadly applicable strategy to simultaneously increase the potency and safety of subunit vaccines.David H. Koch Institute for Integrative Cancer Research at MIT (Koch Institute Support (core) Grant P30-CA14051)National Cancer Institute (U.S.)National Institutes of Health (U.S.) (grant AI091693)National Institutes of Health (U.S.) (grant AI104715)National Institutes of Health (U.S.) (AI095109)United States. Dept. of Defense (contract W911NF-13-D-0001)United States. Dept. of Defense (contract W911NF-07-D-0004)Ragon Institute of MGH, MIT, and Harvar