Ca2+/Calmodulin-Dependent Kinase Kinase α Is Expressed by Monocytic Cells and Regulates the Activation Profile

Macrophages are capable of assuming numerous phenotypes in order to adapt to endogenous and exogenous challenges but many of the factors that regulate this process are still unknown. We report that Ca2+/calmodulin-dependent kinase kinase α (CaMKKα) is expressed in human monocytic cells and demonstrate that its inhibition blocks type-II monocytic cell activation and promotes classical activation. Affinity chromatography with paramagnetic beads isolated an approximately 50 kDa protein from nuclear lysates of U937 human monocytic cells activated with phorbol-12-myristate-13-acetate (PMA). This protein was identified as CaMKKα by mass spectrometry and Western analysis. The function of CaMKKα in monocyte activation was examined using the CaMKKα inhibitors (STO-609 and forskolin) and siRNA knockdown. Inhibition of CaMKKα, enhanced PMA-dependent CD86 expression and reduced CD11b expression. In addition, inhibition was associated with decreased translocation of CaMKKα to the nucleus. Finally, to further examine monocyte activation profiles, TNFα and IL-10 secretion were studied. CaMKKα inhibition attenuated PMA-dependent IL-10 production and enhanced TNFα production indicating a shift from type-II to classical monocyte activation. Taken together, these findings indicate an important new role for CaMKKα in the differentiation of monocytic cells

Phagocytosis of Cholesteryl Ester Is Amplified in Diabetic Mouse Macrophages and Is Largely Mediated by CD36 and SR-A

Type 2 diabetes (T2D) is associated with accelerated atherosclerosis, which accounts for approximately 75% of all diabetes-related deaths. Here we investigate the link between diabetes and macrophage cholesteryl ester accumulation. When diabetic (db/db) mice are given cholesteryl ester intraperitoneally (IP), peritoneal macrophages (PerMΦs) recovered from these animals showed a 58% increase in intracellular cholesteryl ester accumulation over PerMΦs from heterozygote control (db/+) mice. Notably, PerMΦ fluid-phase endocytosis and large particle phagocytosis was equivalent in db/+and db/db mice. However, IP administration of CD36 and SR-A blocking antibodies led to 37% and 25% reductions in cholesteryl ester accumulation in PerMΦ. Finally, in order to determine if these scavenger receptors (SRs) were part of the mechanism responsible for the increased accumulation of cholesteryl esters observed in the diabetic mouse macrophages, receptor expression was quantified by flow cytometry. Importantly, db/db PerMΦs showed a 43% increase in CD36 expression and an 80% increase in SR-A expression. Taken together, these data indicate that direct cholesteryl ester accumulation in mouse macrophages is mediated by CD36 and SR-A, and the magnitude of accumulation is increased in db/db macrophages due to increased scavenger receptor expression

Immunoregulation of bovine macrophages by factors in the salivary glands of Rhipicephalus microplus

<p>Abstract</p> <p>Background</p> <p>Alternative strategies are required to control the southern cattle tick, <it>Rhipicephalus microplus</it>, due to evolving resistance to commercially available acaricides. This invasive ectoparasite is a vector of economically important diseases of cattle such as bovine babesiosis and anaplasmosis. An understanding of the biological intricacies underlying vector-host-pathogen interactions is required to innovate sustainable tick management strategies that can ultimately mitigate the impact of animal and zoonotic tick-borne diseases. Tick saliva contains molecules evolved to impair host innate and adaptive immune responses, which facilitates blood feeding and pathogen transmission. Antigen presenting cells are central to the development of robust T cell responses including Th1 and Th2 determination. In this study we examined changes in co-stimulatory molecule expression and cytokine response of bovine macrophages exposed to salivary gland extracts (SGE) obtained from 2-3 day fed, pathogen-free adult <it>R. microplus</it>.</p> <p>Methods</p> <p>Peripheral blood-derived macrophages were treated for 1 hr with 1, 5, or 10 μg/mL of SGE followed by 1, 6, 24 hr of 1 μg/mL of lipopolysaccharide (LPS). Real-time PCR and cytokine ELISA were used to measure changes in co-stimulatory molecule expression and cytokine response.</p> <p>Results</p> <p>Changes were observed in co-stimulatory molecule expression of bovine macrophages in response to <it>R</it>. <it>microplus </it>SGE exposure. After 6 hrs, CD86, but not CD80, was preferentially up-regulated on bovine macrophages when treated with 1 μg/ml SGE and then LPS, but not SGE alone. At 24 hrs CD80, CD86, and CD69 expression was increased with LPS, but was inhibited by the addition of SGE. SGE also inhibited LPS induced upregulation of TNFα, IFNγ and IL-12 cytokines, but did not alter IL-4 or CD40 mRNA expression.</p> <p>Conclusions</p> <p>Molecules from the salivary glands of adult <it>R. microplus </it>showed bimodal concentration-, and time-dependent effects on differential up-regulation of CD86 in bovine macrophages activated by the TLR4-ligand, LPS. Up regulation of proinflammatory cytokines and IL-12, a Th1 promoting cytokine, were inhibited in a dose-dependent manner. The co-stimulatory molecules CD80, as well as the cell activation marker, CD69, were also suppressed in macrophages exposed to SGE. Continued investigation of the immunomodulatory factors will provide the knowledge base to research and develop therapeutic or prophylactic interventions targeting <it>R. microplus</it>-cattle interactions at the blood-feeding interface.</p

Functional genomics of feeding preferences in Oestroidea

A infestação por larvas de dípteros, conhecida como miíase, é um grande problema na pecuária em todo o mundo e pode causar perdas econômicas severas. A superfamília Oestroidea é um modelo interessante para estudar a evolução de moscas causadoras de miíase devido à diversidade de estratégias de parasitismo entre espécies intimamente relacionadas nessa família. Essas moscas são classificadas pelo seu hábito alimentar como parasitas saprófagos, obrigatórios ou parasitas facultativos. Os parasitas, em particular, podem ser subdivididos pela manifestação clínica/local de infestação como dérmica, nasofaríngea, traumática/ferida e furuncular. Com tal diversidade de estratégias, postula-se que espécies intimamente relacionadas tenham diferenças genéticas que desempenham um papel na formação desses hábitos. Aqui, utilizamos dados de expressão gênica e as sequências codificantes em escala genômica de cinco espécies (Cochliomyia hominivorax, Chrysomya megacephala, Lucilia cuprina, Dermatobia hominis e Oestrus ovis) para encontrar genes que possam estar envolvidos em diferentes estratégias e/ou preferências alimentares. Nós testamos se os 1.287 ortólogos identificados possuiam expressão diferente e restrições evolutivas em diferentes cenários. Ao comparar seus perfis de expressão gênica, encontramos dois genes regulados positivamente; um genes em espécies que causam miíase dérmica envolvido no transporte metabolização de ferro (Ferritina) e outro gene em espécies que causam miíase traumática que responde a níveis reduzidos de oxigênio (anoxia up-regulation-like). Nossa análise evolutiva mostrou um resultado semelhante. Em Ch. hominivorax, encontramos genes diferentes, mas envolvidos nas mesmas funções que podem estar evoluindo sob seleção positiva. Este é o primeiro passo para entender as origens e a evolução da diversidade de estratégias parasitárias em OestroideaThe infestation by dipterous larvae, known as myiasis, is a major problem in livestock worldwide and can cause severe economic losses. The Oestroidea superfamily is an interesting model to study the evolution of myiasis-causing flies because of the diversity of parasitism strategies among closely-related species in this family. These flies are classified by their feeding habit as saprophagous, obligate parasites or facultative parasites. The parasites in particular can be subdivided into dermal, nasopharyngeal, traumatic/wound and furuncular. With such a diversity of parasitic strategies, we expect that closely-related species have genetic differences that play a role in shaping these habits. Here, we used gene expression and coding sequence data from five species (Cochliomyia hominivorax, Chrysomya megacephala, Lucilia cuprina, Dermatobia hominis and Oestrus ovis) to find genes that may be involved in different parasitic strategies. We tested whether 1,287 orthologs have different expression and evolutionary constrains in different scenarios. By comparing their gene expression profiles, we found two up-regulated genes; one in species causing dermic myiasis that is involved in iron transportation/metabolization (Ferritin), and other in species causing traumatic myiasis that responds to reduced oxygen levels (anoxia up- regulated-like). Our evolutionary analysis showed a similar result. In the Ch. Hominivorax branch, we found different genes, but involved in the same functions that may be evolving under positive selection. This is the first step towards understanding the origins and evolution of the diversity of parasitic strategies in Oestroide

Pintura e vertigem

Mais do que investigar algumas das relações possíveis entre a pintura e a fotografia, esta pesquisa tem como objetivo criar uma reflexão sobre a minha produção pictórica que estende-se desde o período da minha graduação e segue adiante, explorando os aspectos que permeiam o trabalho - as questões relativas à história da arte; os parques de diversões sob diferentes aspectos; a imagem automatizada e suas características, especialmente no contexto da tecnologia digital disponível no início do Século XXI.Besides investigating the possible relations between painting and photography, this research work intends on creating a reflection about my pictorical production which began on my graduation years and keeps going on, exploring aspects related to the work - questions related to Art History; amusement parks through their different aspects; automated image and its characteristics, specialy on the context of digital technology available at the beginning of the 21st Centur

Molecular Mechanisms of Promyelocyte Maturation: Implications for Immune Dysfunction in Type II Diabetes

145 p.Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 2003.The data in this thesis indicate one, that the removal of the tyrosine phosphatase CD45 enhances activation of protein kinase C delta (PKCdelta), and two, PKCdelta activity in conjunction with STAT6 regulate the expression of the co-stimulatory marker B7.2 (CD86) and three, the expression of CD86 and the production of cytokines in phorbol-12-myristate-13-acetate (PMA)-stimulated promyelocytes is regulated by calcium calmodulin kinase kinase alpha. These data together indicate that promyelocyte differentiation is a process that is dependent on the activity of PKCdelta. Furthermore, this process is subject to regulation by interlukin-4 induced STAT6 activation and CaMKKalpha. In conclusion we have identified possible targets for future therapies in the treatment of diabetes induced altered immunity.U of I OnlyRestricted to the U of I community idenfinitely during batch ingest of legacy ETD

Advanced computer architectur a design space approach

xxiii.766 : 24 c

Advanced computer architectures : A design space approach

Indeks Bibliografi hlm. Setiap babxxiii, 766 hlm. :il. ;24 cm

Advanced computer architectures :A design space approach

Indeks Bibliografi hlm. Setiap babxxiii, 766 hlm. : il. ; 24 cm