Viral superantigen-induced negative selection of TCR transgenic CD4+ CD8+ thymocytes depends on activation, but not proliferation.

T-cell negative selection, a process by which intrathymic immunological tolerance is induced, involves the apoptosis-mediated clonal deletion of potentially autoreactive T cells. Although different experimental approaches suggest that this process is triggered as the result of activation-mediated cell death, the signal transduction pathways underlying this process is not fully understood. In the present report we have used an in vitro system to analyze the cell activation and proliferation requirements for the deletion of viral superantigen (SAg)-reactive Vbeta8.1 T-cell receptor (TCR) transgenic (TG) thymocytes. Our results indicate that in vitro negative selection of viral SAg-reactive CD4+ CD8+ thymocytes is dependent on thymocyte activation but does not require the proliferation of the negatively signaled thymocytes.

DMC Section: 2008 State of Iowa Juvenile Justice and Delinquency Prevention Act Formula Grant Update, March 2009

This document is the DMC Section of Iowa’s 2009 federal Juvenile Justice and Delinquency Prevention Act (JJDP Act) formula grant three year plan update. The Division of Criminal and Juvenile Justice Planning (CJJP) wrote this update. CJJP is the state agency responsible for administering the JJDP Act in Iowa. Federal officials refer to state administering agencies as the state planning agency (SPA). The Plan was developed and approved by Iowa’s Juvenile Justice Advisory Council. That Council assists with administration of the JJDP Act, and also provides guidance and direction to the SPA, the Governor and the legislature regarding juvenile justice issues in Iowa. Federal officials refer to such state level groups as state advisory groups (SAG’s). The acronyms SPA and SAG are used through this report.

In vivo effects of a recombinant vaccinia virus expressing a mouse mammary tumor virus superantigen.

Early after infection, the mouse mammary tumor virus (MMTV) expresses a superantigen (SAg) at the surface of B lymphocytes. Interaction with the T-cell receptor Vbeta domain induces a polyclonal proliferative response of the SAg-reactive T cells. Stimulated T cells become anergic and are deleted from the T-cell repertoire. We have used a recombinant vaccinia virus encoding the MMTV(GR) SAg to dissect the effects of the retroviral SAg during an unrelated viral infection. Subcutaneous infection with this recombinant vaccinia virus induces a very rapid increase of Vbeta14 T cells in the draining lymph node. This stimulation does not require a large Plumber of infectious particles and is not strictly dependent on the expression of the major histocompatibility complex class II I-E molecule, as it is required after MMTV(GR) infection. In contrast to MMTV infection during which B cells are infected, we do not observe any clonal deletion of the reactive T cells following the initial stimulation phase. Our data show that contrary to the case with MMTV, macrophages but not B cells are the targets of infection by vaccinia virus in the lymph node, indicating the ability of these cells to present a retroviral SAg. The altered SAg expression in a different target cell observed during recombinant vaccinia virus infection therefore results in significant changes in the SAg response.

Fast Track Portland Cement Concrete Pavement, HR-538, 1997

In 1987, 1.5 km (0.935 mi.) of Spruce Hill Drive in Bettendorf, Iowa was reconstructed. It is an arteriel street with commercial usage on both termini with single family residential dwellings along most of the project. A portland cement concrete (PCC) pavement design was selected, but a 14 day curing period would have been an undue hardship on the residents and commercial businesses. An Iowa DOT Class F fast track concrete was used so the roadway could be used in 7 to 10 days. The Class F concrete with fly ash was relatively sticky and exhibited early stiffening problems and substantial difficulty in obtaining the target entrained air content of 6.5%. These problems were never completely resolved on the project. Annual visual field reviews were conducted through 1996. In November 1991, severe premature distress was identified on the westbound two lanes of the full width replacement. The most deteriorated section in a sag vertical, 152 m (500 ft.) of the westbound roadway, was replaced in 1996. Premature distress has been identified on a dozen other conventional PCC Iowa pavements constructed between 1983 and 1989, so the deterioration may not be related to the fact that it was fast track pavement.

MHC class II hierarchy of superantigen presentation predicts efficiency of infection with mouse mammary tumor virus.

Superantigens (SAgs) encoded by infectious mouse mammary tumor viruses (MMTVs) play a crucial role in the viral life cycle. Their expression by infected B cells induces a proliferative immune response by SAg-reactive T cells which amplifies MMTV infection. This response most likely ensures stable MMTV infection and transmission to the mammary gland. Since T cell reactivity to SAgs from endogenous Mtv loci depends on MHC class II molecules expressed by B cells, we have determined the ability of MMTV to infect various MHC congenic mice. We show that MHC class II I-E+ compared with I-E- mouse strains show higher levels of MMTV infection, most likely due to their ability to induce a vigorous SAg-dependent immune response following MMTV encounter. Inefficient infection is observed in MHC class II I-E- mice, which have been shown to present endogenous SAgs poorly. Therefore, during MMTV infection the differential ability of MHC class II molecules to form a functional complex with SAg determines the magnitude of the proliferative response of SAg-reactive T cells. This in turn influences the degree of T cell help provided to infected B cells and therefore the efficiency of amplification of MMTV infection.

Genotyping microarray for CSNB-associated genes.

PURPOSE: Congenital stationary night blindness (CSNB) is a clinically and genetically heterogeneous retinal disease. Although electroretinographic (ERG) measurements can discriminate clinical subgroups, the identification of the underlying genetic defects has been complicated for CSNB because of genetic heterogeneity, the uncertainty about the mode of inheritance, and time-consuming and costly mutation scanning and direct sequencing approaches. METHODS: To overcome these challenges and to generate a time- and cost-efficient mutation screening tool, the authors developed a CSNB genotyping microarray with arrayed primer extension (APEX) technology. To cover as many mutations as possible, a comprehensive literature search was performed, and DNA samples from a cohort of patients with CSNB were first sequenced directly in known CSNB genes. Subsequently, oligonucleotides were designed representing 126 sequence variations in RHO, CABP4, CACNA1F, CACNA2D4, GNAT1, GRM6, NYX, PDE6B, and SAG and spotted on the chip. RESULTS: Direct sequencing of genes known to be associated with CSNB in the study cohort revealed 21 mutations (12 novel and 9 previously reported). The resultant microarray containing oligonucleotides, which allow to detect 126 known and novel mutations, was 100% effective in determining the expected sequence changes in all known samples assessed. In addition, investigation of 34 patients with CSNB who were previously not genotyped revealed sequence variants in 18%, of which 15% are thought to be disease-causing mutations. CONCLUSIONS: This relatively inexpensive first-pass genetic testing device for patients with a diagnosis of CSNB will improve molecular diagnostics and genetic counseling of patients and their families and gives the opportunity to analyze whether, for example, more progressive disorders such as cone or cone-rod dystrophies underlie the same gene defects.

A highly sensitive in vitro infection assay to explore early stages of mouse mammary tumor virus infection.

Mouse mammary tumor virus (MMTV) infection of adult mice induces a strong response to superantigen (Sag) in their draining lymph nodes, which results from the presentation of Sag by MMTV-infected B cells to Sag-reactive T cells. To date, infection with physiologically relevant doses of MMTV can be detected in vivo only after several days of Sag-mediated T-cell-dependent amplification of infected B cells. Furthermore, no efficient in vitro system of detecting MMTV infection is available. Such a model would allow the dissection of the early phase of infection, the assessment of the contributions of different cell types, and the screening of large panels of molecules for their potential roles in infection and Sag response. For these reasons, we have established an in vitro model for detecting infection which is as sensitive and reproducible as the in vivo model. We found that the viral envelope (Env) protein is crucial for target cell infection but not for presentation of Sag. Furthermore, we show that infection of purified B cells with MMTV induces entry of Sag-responsive T cells into the cell cycle, while other professional antigen-presenting cells, such as dendritic cells, are much less efficient in inducing a response.

T cell receptor V beta repertoire in mice lacking endogenous mouse mammary tumor provirus.

When endogenous mouse mammary tumor virus (MMTV) superantigens (SAg) are expressed in the first weeks of life an efficient thymic deletion of T cells expressing MMTV SAg-reactive T cell receptor (TcR) V beta segments is observed. As most inbred mouse strains and wild mice contain integrated MMTV DNA, knowing the precise extent of MMTV influence on T cell development is required in order to study T cell immunobiology in the mouse. In this report, backcross breeding between BALB.D2 (Mtv-6, -7, -8 and -9) and 38CH (Mtv-) mice was carried out to obtain animals either lacking endogenous MMTV or containing a single MMTV locus, i.e. Mtv-6, -7, -8 or -9. The TcR V beta chain (TcR V beta) usage in these mice was analyzed using monoclonal antibodies specific for TcR V beta 2, V beta 3, V beta 4, V beta 5, V beta 6, V beta 7, V beta 8, V beta 11, V beta 12 and V beta 14 segments. Both Mtv-8+ mice and Mtv-9+ mice deleted TcR V beta 5+ and V beta 11+ T cells. Moreover, we also observed the deletion of TcR V beta 12+ cells by Mtv-8 and Mtv-9 products. Mtv-6+ and Mtv-7+ animals deleted TcR V beta 3+ and V beta 5+ cells, and TcR V beta 6+, V beta 7+ and V beta 8.1+ cells, respectively. Unexpectedly, TcR V beta 8.2+ cells were also deleted in some backcross mice expressing Mtv-7. TcR V beta 8.2 reactivity to Mtv-7 was shown to be brought by the 38CH strain and to result from an amino acid substitution (Asn-->Asp) in position 19 on the TcR V beta 8.2 fragment. Reactivities of BALB.D2 TcR V beta 8.2 and 38CH TcR V beta 8.2 to the exogenous infectious viruses, MMTV(SW) and MMTV(SHN), were compared. Finally, the observation of increased frequencies of TcR V beta 2+, V beta 4+ and V beta 8+ CD4+ T cell subsets in Mtv-8+ and Mtv-9+ mice, and TcR V beta 4+ CD4+ T cells in Mtv-6+ and Mtv-7+ mice, when compared with the T cell repertoire of Mtv- mice, is consistent with the possibility that MMTV products contribute to positive selection of T cells.

Role of dendritic cells in the immune response induced by mouse mammary tumor virus superantigen.

After mouse mammary tumor virus (MMTV) infection, B lymphocytes present a superantigen (Sag) and receive help from the unlimited number of CD4(+) T cells expressing Sag-specific T-cell receptor Vbeta elements. The infected B cells divide and differentiate, similarly to what occurs in classical B-cell responses. The amplification of Sag-reactive T cells can be considered a primary immune response. Since B cells are usually not efficient in the activation of naive T cells, we addressed the question of whether professional antigen-presenting cells such as dendritic cells (DCs) are responsible for T-cell priming. We show here, using MMTV(SIM), a viral isolate which requires major histocompatibility complex class II I-E expression to induce a strong Sag response in vivo, that transgenic mice expressing I-E exclusively on DCs (I-EalphaDC tg) reveal a strong Sag response. This Sag response was dependent on the presence of B cells, as indicated by the absence of stimulation in I-EalphaDC tg mice lacking B cells (I-EalphaDC tg muMT(-/-)), even if these B cells lack I-E expression. Furthermore, the involvement of either residual transgene expression by B cells or transfer of I-E from DCs to B cells was excluded by the use of mixed bone marrow chimeras. Our results indicate that after priming by DCs in the context of I-E, the MMTV(SIM) Sag can be recognized on the surface of B cells in the context of I-A. The most likely physiological relevance of the lowering of the antigen threshold required for T-cell/B-cell collaboration after DC priming is to allow B cells with a low affinity for antigen to receive T-cell help in a primary immune response.

In vitro negative selection of viral superantigen-reactive thymocytes by thymic dendritic cells.

Intrathymic expression of endogenous mouse mammary tumor virus (MMTV)-encoded superantigens (SAg) induces the clonal deletion of T cells bearing SAg-reactive T-cell receptor (TCR) Vbeta elements. However, the identity of the thymic antigen-presenting cells (APC) involved in the induction of SAg tolerance remains to be defined. We have analyzed the potential of dendritic cells (DC) to mediate the clonal deletion of Mtv-7-reactive TCR alphabeta P14 transgenic thymocytes in an in vitro assay. Our results show that both thymic and splenic DC induced the deletion of TCR transgenic double positive (DP) thymocytes. DC appear to be more efficient than splenic B cells as negatively selecting APC in this experimental system. Interestingly, thymic and splenic DC display a differential ability to induce CD4+ SP thymocyte proliferation. These observations suggest that thymic DC may have an important role in the induction of SAg tolerance in vivo.

Microgrids and their operations

Interconnection of loads and small size generation forms a new type of distribution systems, the Microgrid. The microgrids can be operated together with the utility grid or be operated autonomously in an island. Thesesmall grids present a new paradigm of the construction of the low voltage distribution systems. The microgrids in the distribution systems can become small, controllable units, which immediately react to the system's changes. Along with that the microgrids can realize the special properties, such as increasing the reliability, reducing losses, voltage sag correction, uninterruptible supplying. The goals of the thesis are to explain the principles of the microgrid's functioning, to clarify the main ideas and positive features of the microgrids, to find out and prove their advantages and explain why they are so popular nowadays all over the world. The practical aims of the thesis are to construct and build a test setup of a microgrid based on two inverters from SMA Technologie AG in the laboratory and to test all the main modes and parameters of the microgrid's operating. Also the purpose of the thesis is to test the main component of the microgrid - the battery inverter which controls allthe processes and energy flows inside a microgrid and communicates with the main grid. Based on received data the main contribution of the thesis consists of the estimation of the established microgrid from the reliability, economy and simplicity of operating points of view and evaluation ofthe advisability of its use in different conditions. Moreover, the thesis assumes to give the recommendations and advice for the future investigations of the built system.

Metodologias de análise de cadeias agroindustriais: aplicação para citros

Este artigo teve como objetivo apresentar os principais resultados de um estudo de Sistemas Agroindustriais (SAG's) do PENSA e os principais resultados de sua aplicação à citricultura no Brasil. Este é um modelo utilizado para descrever diversas etapas metodológicas e quantificar em termos de faturamento os principais setores do agronegócio. A proposta visa a manter o objetivo que norteou o desenvolvimento do trabalho. No SAG dos citros foram identificadas as principais empresas privadas e instituições envolvidas, desde as indústrias de insumos, à citricultura, passando pela indústria processadora de citros, distribuição , até chegar ao consumidor final. As informações foram coletadas através de entrevistas, ponto central desta metodologia, buscando levantar o montante financeiro vendido pelas empresas no setor em estudo. Neste ponto, os dados obtidos são processados e inseridos na descrição do sistema (quantificação) e assim analisados. Foi gerado o desenho da cadeia de cítricos objetivando identificar possíveis ações coletivas, visando ao fortalecimento do setor.

Syöttöjännitteen jännitehäiriöt paperikoneen sähkökäytössä

Diplomityön tavoitteena oli selvittää paperikonelinjakäyttöjen toimintaa syöttöjännitteen alenemien ja –jännitekatkojen aikana. Työssä haluttiin selvittää, millaisista häiriöistä laitteistot selviävät pysähtymättä ja kuinka laitteistojen immuniteettia häiriöille voitaisiin parantaa. Standardien esittämien jännitehäiriöiden siedon vaatimuksien lisäksi haluttiin selvittää olosuhteet, joissa asiakkaiden sähkökäytöt tulisi toimia. Näiden ongelmien ratkaisemiseksi selvitettiin laboratoriomittauksin tyristori- ja ISU –syöttöyksiköillä varustettujen linjakäyttöjen testilaitteistojen ominaisuuksia jännitteen alenemien aikana. Lisäksi selvitettiin paperikoneen muiden laitteistojen toimintaa jännitehäiriöiden aikana. Avain sähkökäyttöjen jännitehäiriöiden siedon parantamiseksi löytyy ulkoisista energiavarastoista ja paperikoneen linjakäyttöjen hallitusta ohjaamisesta häiriöiden aikana. Näiden hyödyntäminen edellyttää kuitenkin työssä selvitetyn paperiprosessin ohjauksen tuntemista ratakatkon aikana.

Jännitteenkorotin heikkoihin jakeluverkkoihin

Sähkönkulutuksen kasvaessa jakeluverkossa syntyy yhä enemmän jännitteenalenemia, jotka ylittävät sähkön laatustandardin asettamat raja-arvot. Jännitteenalenemista kärsivät etenkin syrjäseuduilla asuvat ihmiset, joiden käyttökohteet sijaitsevat kaukana jakelumuuntajasta. Perinteinen tapa on ollut kasvattaa syöttävän jakelukaapelin poikkipinta-alaa ja vaihtaa jakelumuuntaja suurempaan ongelman ratkaisemiseksi, mutta investointi on kuitenkin kallis eikä välttämättä maksa itseään koskaan takaisin. On myös mahdollista, että aiempaa suuremman muuntajan tyhjäkäyntihäviöillä on huonontava vaikutus järjestelmän energiatehokkuuteen. Tällöin voi olla kokonaistaloudellisesti kannattavampaa liittää sähköverkkoon laite, jolla voidaan korjata jännitteenalenemia. Diplomityössä tutkitaan säästömuuntajaan perustuvaa jännitteenkorotinta, jolla voidaan vähentää jakeluverkon jännitteenalenemien aiheuttamia haittoja. Työssä tutkitaan laitteen eri kytkintopologioita sekä niiden ohjausmenetelmiä, joilla voidaan muuttaa säästömuuntajan käämikierrosmääriä sen ollessa kuormitettuna.

Estudo endócrino reprodutivo e do comportamento sócio-sexual de sagui-de-tufo-preto (Callithrix penicillata) mantido em cativeiro

A comunicação do estado reprodutivo nos primatas da família Callithrichidae, depende principalmente dos comportamentos sócio-sexuais como um sistema de sinalização primário, uma vez que nestas espécies a ovulação não é percebida pelos machos. Neste trabalho, os padrões de comportamentos sócio-sexuais foram analisados em conjunto com as concentrações de metabólitos fecais dos esteróides sexuais progesterona (MFP), estradiol (MFE) e testosterona (MFT) em casais cativos de Sagüi-de-tufos-pretos (Callithrix penicillata), nas diferentes fases do ciclo ovariano. O grupo estudado era composto por quarto casais adultos, mantidos no Centro de Reabilitação de Animais Selvagens da prefeitura de São Paulo. Os padrões comportamentais foram registrados pelo método de amostragem focal por intervalo de tempo a cada 30 segundos, cinco vezes por semana, totalizando 14.400 registros por animal. A mensuração das concentrações de metabólitos fecais dos esteroides sexuais foram realizados pelo método de enzima imunoensaio (EIE). Os resultados obtidos dessas concentrações possibilitaram a determinação endócrina das fases do ciclo ovariano (folicular e luteal) e de suas respectivas durações, assim como a determinação da fase periovulatória. Foram caracterizados 31 ciclos ovarianos completos, com duração de 24,3±4,1 dias (média ±DP), sendo que a fase folicular compreendeu 13,04±4,8dias e a fase lútea 11,2±4,2 dias. Os comportamentos sócio-sexuais (marcação por cheiro, cheirar genitália, catação e apresentação sexual) e a variável "proximidade" mostraram-se significativamente mais prevalentes na fase periovulatória do que nas demais fases do ciclo. Não houve alteração das concentrações de MFT dos machos ao longo de todo o período estudado. A análise conjunta das concentrações de metabólitos fecais de esteróides sexuais e dos comportamentos sócio-sexuais possibilitou um melhor entendimento das relações endócrino-comportamentais e reprodutivas de C. penicillata.