The adenylate energy charge values of twenty bacteria isolated from soil and cultured in a medium consisting of soil and distilled water were determined by the luciferin-luciferase bioluminescense method. The purpose of this study was to examine the growth and energy charge values of these organisms in soil extract medium, and to determine what effect the addition of glucose has on their energy charge values. Three of the organisms employed in this study showed energy charge values similar to those reported for bacteria grown in enriched media. The remainder of the isolates demonstrated low energy charge values, and scant growth in the soil medium.

The plasma membrane calcium-transport ATPase plays a major role in maintaining the low cytosolic calcium concentrations required for normal cellular function. Calcium, magnesium, calmodulin and lanthanum have been shown to alter the activity of the calcium-stimulated, magnesium-dependent ATPase activity in human erythrocytes. In an attempt to examine the reaction sequence of the (Ca²⁺ + Mg²⁺)-ATPase, the effects of these agents on the kinetics of calcium dependent phosphoprotein formation, the first step in the partial reaction sequence, were examined. Calmo-dulin-depleted erythrocyte membranes were prepared by hypotonic lysis in the presence of EDTA, according to the method of Carafoli et al (1980). Calcium-dependent formation of the phosphorylated intermediate was biphasic; the high calcium-affinity component was associated with low levels of E.Ca.P and a shallow response to changing calcium concentrations, whereas in the region of the low calcium-affinity component, E.Ca.P rose sharply in response to increasing calcium concentrations. The low affinity component of E.Ca.P lies in the range of calcium concentrations which inhibit (Ca²⁺ + Mg²⁺)-ATPase activity. When analyzed on LiDS acid PAGE, both components of calcium-dependent phosphoprotein formation were due to hydroxylamine-sensitive phosphorylation of a 135,000-145,000 dalton protein. Hence, the low calcium-affinity component of phosphoprotein formation and calcium-dependent inhibition of (Ca²⁺ + Mg²⁺)-ATPase activity were likely due to calcium-inhibition of dephosphorylation. Kinetic studies of calcium-dependent phosphoprotein formation, at two different calcium concentrations (1.0 μM, 0.4 mM), indicated that a steady-state was reached much sooner at higher calcium concentrations. Lanthanum, which is known to block dephosphorylation of the intermediate complex, increased both the apparent rate of formation and the steady-state level of the phosphorylated intermediate. Calmodulin, which has previously been shown to increase both the maximum velocity and the calcium affinity of the (Ca²⁺ + Mg²⁺)-ATPase, did not affect either calcium-dependent inhibition of (Ca²⁺ + Mg²⁺ )-ATPase activity or the biphasic nature of calcium-dependent phosphoprotein formation. At low calcium concentrations, calmodulin increased the apparent rate of phosphoprotein formation, whereas at higher calcium concentrations (0.4 mM) calmodulin reduced the steady-state level of the phosphoprotein; the apparent rate of formation was unaffected. In the presence of lanthanum, calmodulin increased both the apparent rate of formation and steady-state level of the phosphoprotein, suggesting that the true rate of formation was increased by calmodulin at higher calcium concentrations, but this was normally hidden by a simultaneous increase in the rate of dephosphorylation. Removal of endogenous magnesium, using trans-1,2-diamino-cyclohexane tetraacetic acid (CDTA) did not alter the calcium sensitivity or rate of formation of the phosphorylated intermediate, however turnover of the intermediate was markedly reduced. In the absence of free magnesium, both the velocity and calcium sensitivity of the (Ca²⁺ + Mg²⁺)-ATPase were also found to be lower. The low calcium-affinity component of calcium-dependent phosphoprotein formation, which Schatzmann (1982) has attributed to an action of calcium at a "magnesium-specific" site, was not affected by magnesium concentrations as high as 1 mM. Furthermore, this phosphoprotein could be dephosphorylated along either the forward or reverse pathways. These results indicate that the transformation from E₁.Ca.P to E₂.Ca.P may not be the site of the calcium-dependent inhibition of dephosphorylation. Calmodulin-depleted membrane fragments were prepared from the erythrocytes of cystic fibrosis patients as well as age- and sex-matched controls. Under conditions in which dephosphoryla-tion is inhibited, phosphoprotein formation and (Ca²⁺ + Mg²⁺)-ATPase activities were determined. Both (Ca²⁺ + Mg²⁺)-ATPase activity and phoshoprotein formation were found to be significantly reduced in the preparations derived from patients with cystic fibrosis. Turnover of the phosphorylated intermediate did not differ significantly between the two groups. A reduction in (Ca²⁺ + Mg²⁺)-ATPase activity and phosphoprotein formation suggests that there may be fewer active calcium-pumping sites in the erythrocyte membranes of cystic fibrosis patients compared to normal subjects. / Pharmaceutical Sciences, Faculty of / Graduate

The role of cyclic AMP and a differentiation-indueing factor (DIF) in the differentiation of stalk cells was investigated in the cellular slime mold Dictyostelium discoideum. In this organism, starvation triggers the aggregation of amoebae into multicellular masses within which a simple, well-regulated pattern of partially differentiated cells is formed and which ultimately form fruiting bodies comprised of spore and stalk cells. In a monolayer system at low cell densities, stalk cell formation is dependent on the presence of both cyclic AMP and DIF. Both factors act within a short time of each other, induction by cyclic AMP preceding induction by DIF, beginning between 8 to 10 hours of incubation in monolayers, and progressively committing an increasing proportion of the cells in monolayer to form stalk cells. The relative effectiveness of analogues of cyclic AMP to induce stalk cell formation in monolayers indicates that the well-characterized cell surface cyclic AMP receptor most probably mediates the action of cyclic AMP. Although this receptor appears early during aggregation, it does not become activated until later during development in vivo, probably because the cyclic AMP concentrations within developing cell masses must build up to levels higher than those in aggregation streams. The finding that caffeine inhibits stalk cell formation in low density monolayers and that the permeable analogue 8-Bromo-cyclic AMP can partially reverse this inhibition suggests that activation of this receptor leads to an increase in internal cyclic AMP levels as one of the steps in stalk cell differentiation. The finding that the expression in low density monolayers of AP IV, a cell-type non-specific isozyme of acid phosphatase, was cyclic AMP-dependent is consistent with the view that cyclic AMP induces non-specific postaggregative gene expression during development in vivo. The findings that the expression of pre-stalk arid stalk cell specific antigens and of the pre-stalk cell specific isozyme AP II was DIF-dependent provide good evidence for the idea that both pre-stalk and stalk cell formation are induced by DIF. The fact that isolated pre-stalk cells require DIF for stalk cell formation in low density monolayers further supports this idea. Whereas cells independent of DIF for stalk cell formation in monolayers appear immediately after cyclic AMP-independent cells during differentiation in low density monolayers, DIF-independent cells appear considerably later during development in vivo. This evidence and the fact that developing cell masses contain elevated levels of DIF lead to the postulate that the factor(s) which triggers the formation of fruiting bodies also controls the pre-stalk to stalk cell conversion. / Science, Faculty of / Botany, Department of / Zoology, Department of / Graduate

The mechanism of regulation of the Ca²⁺ pump ATPase of the human erythrocyte membrane by calmodulin, cyclic AMP and the anion channel was studied using membrane fragments, resealed "ghosts", inside-out vesicles and a Triton X-100 solubilized enzyme preparation. The (Ca²⁺ + Mg²⁺ )-ATPase activity in erythrocyte membranes or a Triton X-100 solubilized enzyme preparation showed biphasic (high and low affinity) Ca²⁺ activation kinetics. The anionic calcium binding protein, calmodulin, increased both the calcium sensitivity (Kca²⁺) and the maximum velocity (Vmax ) of the enzyme. Certain polyanionic agents (poly-L-aspartic acid, poly-L-glutamic acid), alicyclic sulfonic acids (HEPES,N-2-hydroxyethylpiperazine-N¹-2-ethanesulfonic acid, MES,2-N- (morpholinoethanesulfonic acid)), and aromatic carboxylic acids (benzoic and salicylic acids) increased the Kca²⁺ but not the Vmax of (Ca²⁺ + Mg²⁺ )-ATPase in erythrocyte membranes and Triton X-100 solubilized enzyme preparations. Trifluoperazine (30 μM) antagonized activation of the enzyme by calmodulin and poly-L-aspartic acid, but not by sodium-HEPES or sodium-MES. Limited trypsin proteolysis of (Ca²⁺ + Mg²⁺ )-ATPase in the erythrocyte membrane abolished activation by calmodulin, poly-L-aspartic acid and sodium-HEPES. These results suggest that the modulation of the Ca²⁺ sensitivity of (Ca²⁺ + Mg²⁺ )-ATPase by calmodulin may be associated with the anionic properties of this protein, and that this property can be mimicked by some other anions, probably by interacting at an anion-regulatory site on the enzyme. Cyclic AMP (5 μM) was found to inhibit the (Ca²⁺ + Mg²⁺)-ATPase activity (approx. 20%) in erythrocyte membranes, probably via endogenous cyclic AMP protein kinase, since this effect could be blocked by cyclic AMP protein kinase inhibitor (PKI) from the rabbit skeletal muscle, By contrast, bovine heart PKI stimulated (Ca²⁺ + Mg²⁺ )-ATPase activity (approx. 100%) by increasing the Kca²⁺ but not the Vmax of the enzyme in membrane or Triton X-100 solubilized preparations. At a low calcium concentration the stimulation by bovine heart PKI and saturating levels of calmodulin was additive, suggesting that the two effectors acted by distinct mechanisms. The stimulation of (Ca²⁺ + Mg²⁺ )-ATPase activity by bovine heart PKI was not solely due to its antagonism of the protein kinase because a) modification of arginine residues of bovine heart PKI abolished its inhibition of cyclic AMP protein kinase, but had no effect on the stimulation of (Ca²⁺ + Mg²⁺ )-ATPase; b) trifluoperazine (20 μM) antagonized the stimulation of (Ca²⁺ + Mg²⁺ )-ATPase by PKI, similarly to its antagonism of calmodulin stimulation, but it did not affect the inhibition of protein kinase by PKI. It is suggested that different mechanisms are involved in the inhibition of cyclic AMP protein kinase and the stimulation of (Ca²⁺ + Mg²⁺ )-ATPase by bovine heart cyclic AMP PKI. Next, the role of anion channel blockers on the (Ca²⁺ + Mg²⁺ )- ATPase was studied. The photolabeling reagent N-(4-azido-2-nitrophenyl)- 2 aminoethylsulfonate (NAP-taurine) was found to inhibit the (Ca²⁺+ Mg²⁺ )-ATPase of fragmented red cell membranes. Half maximal inhibition occurred between 25 μM and 50 μM. At these concentrations Mg²⁺ -ATPase and (Na⁺ + K⁺)-ATPase activities in the membranes were not affected. The reversible inhibition of (Ca²⁺ + Mg²⁺ )-ATPase produced by NAP-taurine in the dark became irreversible after photolysis in the presence of this reagent. Incubation of the membranes with Ca²⁺ , Mg²⁺ , ATP or calmodulin, prior to photolysis in the presence of NAP-taurine, did not protect the enzyme from Inhibition. Limited trypsin proteolysis of (Ca²⁺ + Mg²⁺ )-ATPase in fragmented membranes, which abolished activation by calmodulin, did not affect the inhibition by NAP-taurine. NAP-taurine was found to Inhibit the (Ca²⁺ + Mg²⁺ )-ATPase activity from the cytoplasmic side of the membrane, as determined from the following experiments. Addition of NAP-taurine (50 μM) to resealed erythrocyte ghosts inhibited less than 5% of the (Ca²⁺ + Mg²⁺ )-ATPase activity, compared to 50-60% Inhibition in ghosts resealed in the presence of 50 μM NAP-taurine. Furthermore, NAP-taurine inhibited ATP-dependent Ca²⁺ - transport into inside-out vesicles at a similar concentration (50 μM). The inhibition of the (Ca²⁺ + Mg²⁺ )-ATPase activity of membranes by NAP-taurine appeared to be a direct action on the enzyme, rather than through inhibition of the anion channel, as (Ca²⁺ + Mg²⁺ )-ATPase activity was not inhibited in membranes made from red blood cells reacted irreversibly with 50 μM NAP-taurine or the anion channel blocker 4,4'-diisothiocyano- 2,2' stilbene disulfonate (DIDS) (5 μM) or in membranes assayed in the presence of another anion channel blocker, probenecid (125 μM). This is the first reported selective antagonist of the Ca²⁺ pump, and it is suggested that NAP-taurine could be a useful tool for studying the Ca²⁺- transport ATPase in a variety of cells. / Pharmaceutical Sciences, Faculty of / Graduate

As células T regulatórias (Tregs) são essenciais para a manutenção da tolerância periférica, prevenção de doenças autoimunes e limitantes nas doenças inflamatórias crônicas. Além disso, essas células exercem um papel fundamental no controle da rejeição de transplantes. Diferentes protocolos mostraram que é possível obter Tregs a partir de células T naive CD4+ in vitro. Para tal, é consenso que o TGF-? e a interleucina-2 (IL-2) são capazes de direcionar as células T naive CD4+ a se tornarem regulatórias após um estímulo antigênico (anti-CD3/CD28). Nosso grupo recentemente notou que, durante a imunomodulação de linfócitos T pelas células estromais mesenquimais (CTMs), estas eram capazes de produzir adenosina que, por sua vez, participa do processo de imunorregulação. Outros trabalhos indicam que as CTMs suprimem a proliferação dos linfócitos T pela geração de Tregs e que as CTMs induzem a geração de Tregs através da regulação negativa da via TCR e da via AKTmTOR. Evidências apontam que a adenosina pode atuar regulando negativamente a via mTOR. Portanto, acredita-se que a adenosina possa participar do processo de geração de Tregs através da modulação da via mTOR. Além disso, estudos recentes indicam que a ativação de receptores de adenosina, mais especificamente A2a, com agentes agonistas, leva ao aumento da produção de células Tregs, enquanto que a utilização de agentes antagonistas destes receptores leva à diminuição da diferenciação de Tregs. Porém, estes estudos mostram a geração de Tregs a partir de células T naive de camundongos. Visto a grande importância das Tregs no contexto imunológico, a produção eficiente de Tregs in vitro tem importância fundamental para o desenvolvimento de novos protocolos terapêuticos para o tratamento de doenças autoimunes e no combate à rejeição de transplantes. Assim, o objetivo central deste trabalho foi avaliar a participação de agonistas e antagonistas de receptores de adenosina na indução de células T regulatórias geradas in vitro (iTreg) pela ativação de células T CD4+ naive isoladas de sangue de cordão umbilical (SCU) humano. Para isso, células mononucleares foram isoladas de bolsas de SCU e as células T naive foram isoladas imunomagnéticamente. Essas células foram ativadas com beads ligadas a anticorpos anti-CD2/CD3/CD28 e cultivadas por cinco dias na presença de IL-2 e diferentes concentrações de drogas agonistas e antagonistas de receptores de adenosina. Em seguida, foram avaliados os principais marcadores de células T regulatorias por meio de citometria de fluxo e o meio de cultura foi coletado ao final da geração para quantificação de citocinas. Além disso, o RNA total foi extraído de todas as condições de cultivo para a análise da expressão de genes envolvidos na geração e desenvolvimento das Tregs, por PCR quantitativo. O potencial de supressão de células T efetoras também foi avaliado. / Regulatory T cells (Tregs) are essential for the maintenance of peripheral tolerance, prevention of autoimmune and limiting diseases in chronic inflammatory diseases. In addition, these cells play a key role in the control of transplant rejection. Different protocols have shown that it is possible to obtain Tregs from naive CD4+ T cells in vitro. To this end, there is consensus that TGF-? and interleukin-2 (IL-2) are capable of directing the naive CD4 + T cells to become regulatory following an antigenic stimulus (anti-CD3/CD28).. Our group recently noted that during the immunomodulation of T lymphocytes by mesenchymal stromal cells (MSCs), they were able to produce adenosine which in turn participates in the immunoregulation process. Other studies indicate that MSCs suppress the proliferation of T lymphocytes by generation of Tregs and that MSCs induce generation of Tregs by downregulation of the TCR pathway and the AKT-mTOR pathway. Evidence indicates that adenosine may act by downregulating the mTOR pathway. Therefore, it is believed that adenosine may participate in the generation of Tregs by modulating the mTOR pathway. In addition, recent studies indicate that activation of adenosine receptors, more specifically A2a, with agonist agents, leads to increased production of Treg cells, whereas the use of antagonistic agents of these receptors leads to a decrease in Treg differentiation.. However, these studies show the generation of Tregs from naive T cells of mice. In view of the great importance of Tregs in the immunological context, the efficient production of Tregs in vitro is of fundamental importance for the development of new therapeutic protocols for the treatment of autoimmune diseases and in the fight against transplant rejection. Thus, the central objective of this study was to evaluate the participation of adenosine receptor agonists and antagonists in induction of regulatory T cells generated in vitro (iTreg) by the activation of naive CD4+ T cells isolated from human umbilical cord blood (SCU). For this, mononuclear cells were isolated from SCU and naive T cells were immunomagnetic isolated. These cells were activated with beads bound to anti-CD2/CD3/CD28 antibodies and cultured for five days in the presence of IL-2 and different concentrations of agonist drugs and antagonists of adenosine receptors. Next, the major regulatory T-cell markers were assessed by flow cytometry and the culture medium was collected at the end of the generation for quantification of cytokines. In addition, total RNA was extracted from all culture conditions for the analysis of the expression of genes involved in the generation and development of Tregs by quantitative PCR. The potential for suppression of effector T cells was also evaluated.

Background Efficacy and safety of treatments for hospitalized COVID-19 are uncertain. We systematically reviewed efficacy and safety of remdesivir for the treatment of COVID-19. Methods Studies evaluating remdesivir in adults with hospitalized COVID-19 were searched in several engines until August 21, 2020. Primary outcomes included all-cause mortality, clinical improvement or recovery, need for invasive ventilation, and serious adverse events (SAEs). Inverse variance random effects meta-analyses were performed. Results We included four randomized controlled trials (RCTs) (n = 2296) [two vs. placebo (n = 1299) and two comparing 5-day vs. 10-day regimens (n = 997)], and two case series (n = 88). Studies used intravenous remdesivir 200mg the first day and 100mg for four or nine more days. One RCT (n = 236) was stopped early due to AEs; the other three RCTs reported outcomes between 11 and 15 days. Time to recovery was decreased by 4 days with remdesivir vs. placebo in one RCT (n = 1063), and by 0.8 days with 5-days vs. 10-days of therapy in another RCT (n = 397). Clinical improvement was better for 5-days regimen vs. standard of care in one RCT (n = 600). Remdesivir did not decrease all-cause mortality (RR 0.71, 95% CI 0.39 to 1.28, I2 = 43%) and need for invasive ventilation (RR 0.57, 95%CI 0.23 to 1.42, I2 = 60%) vs. placebo at 14 days but had fewer SAEs; 5-day decreased need for invasive ventilation and SAEs vs. 10-day in one RCT (n = 397). No differences in all-cause mortality or SAEs were seen among 5-day, 10-day and standard of care. There were some concerns of bias to high risk of bias in RCTs. Heterogeneity between studies could be due to different severities of disease, days of therapy before outcome determination, and how ordinal data was analyzed. Conclusions There is paucity of adequately powered and fully reported RCTs evaluating effects of remdesivir in hospitalized COVID-19 patients. Until stronger evidence emerges, we cannot conclude that remdesivir is efficacious for treating COVID-19. / Revisión por pares

<p>  </p> <p>Adenosine-to-Inosine (A-to-I) mRNA editing is a post-transcriptional modification of specific sites within the mRNA that has only recently been observed in filamentous fungi. In the wheat scab fungus <em>Fusarium graminearum,</em> this phenomenon has shown to be facilitated by FgTad2 and FgTad3, homologs of Adenine Deaminase Acting on tRNA (ADAT). Interestingly, these two proteins are constitutively expressed in all different life stages<em>, </em>in contrast to only the sexual stage-specific nature of A-to-I mRNA editing in <em>F. graminearum</em>. To understand the molecular mechanisms regulating this process, six candidate co-factor genes were identified which interact with FgTad2 and/or FgTad3, specifically during sexual reproduction. Deletion mutants of four candidate co-factor genes were successfully generated. All four mutants displayed normal asexual development of <em>F. graminearum</em>, but four mutants also altered sexual function. Those four mutant led to formation of morphologically normal perithecia and ascospores, but the perithecia failed to discharge ascospores. More interestingly, in <em>FGSG_10943 </em>deletion mutant, most of these ascospores germinated precociously within the perithecium. I also observed, that among the candidate co-factor genes which are specifically expressed during sexual reproduction, <em>FGSG_10943</em> was significantly upregulated during the later stage of sexual development. This gene is restricted in nature to only a few orders of fungi in the class Sordariomycetes that form dark pigmented ascocarps, particularly Hypocreales and Glomerellales. Taken together, these results indicate that the four candidate co-factor genes are dispensable for vegetative growth of the fungus and involved in ascospore discharge. <em>FGSG_10943</em> appears to be involved in autoinhibition of ascospores inside the perithecia and interact with FgTad2 during sexual reproduction to mediate A-to-I mRNA editing in <em>F. graminearum</em>.</p>

No description available.

To determine the mechanism by which AMP 579, an adenosine A1/A2 agonist, administered at reperfusion protects ischemic myocardium, buffer-perfused rabbit hearts were subjected to 30 min of global ischemia and 2 h of reperfusion. AMP 579 (500 nM) was included in the reperfusate for the first 70 min. Average left ventricular diastolic pressure during reperfusion in hearts receiving AMP 579 was lower than that in control hearts (17.9 ± 2.4 vs. 39.0 ± 6.5 mm Hg, p < 0.05), indicating attenuation of contracture. Left ventricular developed pressure and coronary flow during reperfusion were also significantly improved with AMP 579 treatment. AMP 579's anti-contracture effect was blocked by the adenosine A2-receptor antagonist 8-(3-chlorostyryl)caffeine (CSC), but not by the A1 antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX). CSC, but not DPCPX, also blocked AMP 579's ability to preserve developed pressure and coronary flow in these hearts. AMP 579 significantly reduced infarction in isolated hearts subjected to regional ischemia. The anti-infarct effect again was abolished by CSC but not by DPCPX. Finally, we tested whether 5′-(N-ethylcarboxamido)adenosine (NECA), another A1/A2 agonist, also administered for the initial 70 min of reperfusion, could duplicate the anti-infarct effect of AMP 579. One-hundred-nanomolar NECA duplicated the protection, but neither 50 nM CGS21680, a selective A2 agonist, nor 100 μM adenosine was protective. Therefore, AMP 579 given at reperfusion reduces contracture and infarction. Anti-contracture and anti-infarct effects require the adenosine A2, but not the A1, receptor suggesting that prevention of contracture and tissue salvage are mechanistically related. Not all A2 agonists were able to duplicate the anti-infarct effect, suggesting something unique about AMP579.

Obesity is a significant health care problem, affecting more than one third of the United States population and is an important risk factor for Type 2 Diabetes Mellitus (T2D). Adipose tissue expansion results in the recruitment and accumulation of macrophages, which secrete proinflammatory cytokines that impair insulin signaling. Adenosine regulates inflammation by signaling through G-protein coupled receptors (GPCRs), such as the A2b adenosine receptor (A2bAR). Recently a role for adenosine receptors has been described in the differentiation of osteoblasts and adipocytes. This thesis tests the hypothesis that the A2bAR regulates adipose tissue dynamics at the level of preadipocyte differentiation and macrophage inflammation. This thesis showed that activation of the A2bAR inhibited preadipocyte differentiation. A2bAR-induced adipocyte inhibition was dependent on the expression of Krüppel-like factor 4 (KLF4), which is important for stem cell maintenance and renewal. A2bAR knockdown enhanced adipogenesis in vitro and A2bAR knockout (KO) mice had more adipocytes as compared to wild type (WT) mice, suggesting enhanced adipogenesis in the absence of the A2bAR. The translational potential of this work is strengthened by the previous finding of elevated A2bAR expression in adipose tissue of obese individuals as well as our new finding of a close correlation between the expression of A2bAR and KLF4 in adipose tissue of obese individuals. A2bAR KO mice have impaired insulin resistance, in part due to reduced levels of insulin receptor substrate-2 (IRS-2). Proinflammatory cytokines have been shown to reduce IRS-2 levels. Given the role of the A2bAR in regulating inflammation, the contribution of A2bAR signaling in macrophages to insulin resistance was elucidated. Transgenic mice that express A2bAR only in macrophages were generated. Intriguingly, restoration of A2bAR signaling in macrophages ameliorated insulin resistance, glucose tolerance, and fat and liver tissue insulin signaling. As expected, tissue and plasma proinflammatory cytokine levels were reduced to that of WT mice. This suggested that the protective effect of A2bAR signaling on insulin resistance was due in large part to A2bAR control of macrophage cytokine expression. This thesis highlights the importance of A2bAR signaling in adipogenesis and in regulating inflammation in the setting of obesity and T2D.