Human studies evaluating dGEMRIC as a prognostic tool for knee osteoarthritis

Osteoarthritis (OA) is the most common joint disorder worldwide, causing joint pain and stiffness. The current gold standard for diagnosing knee OA is radiography. However, the disease has often progressed well beyond the point of no return once radiographic cartilage changes become visible. Identifying changes in cartilage at an early stage of OA would allow curative or prophylactic treatment to be instigated much earlier than today. Early in the progression of the disease, the articular cartilage is depleted of glycosaminoglycans (GAGs), which are responsible for cartilage load distribution and compressive stiffness. Delayed gadolinium-enhanced magnetic resonance imaging of cartilage, abbreviated dGEMRIC, can be used to estimate the GAG content of cartilage. A contrast medium is used, the concentration of which in the cartilage is inversely proportional to the amount of GAG. The purpose of this work was to evaluate dGEMRIC as a prognostic tool for knee cartilage changes and knee OA in humans. It was found that dGEMRIC could be used to predict the development of radiographic knee OA in patients at risk of developing OA. An association was also found between dGEMRIC values and important features of knee OA, such as joint space narrowing and osteophytes (bony changes). Unloading of joints has previously been shown to affect the constituents of cartilage. The knees of patients with ankle fractures, prescribed unloading of the injured leg for six weeks, were investigated. Unloading resulted in a measurable effect on the constituents of the knee cartilage, seen as a decrease in GAG content and an increase in the range of dGEMRIC values. These findings should be taken into account when considering treatment of patients involving an unloading regimen. Anterior cruciate ligament (ACL) injury has previously been shown to be an important risk factor for the development of OA. Patients who had sustained an ACL injury 20 years earlier, who had not undergone ACL reconstruction, were investigated. Notably, these patients showed good cartilage quality and subjective knee function, similar to that in healthy reference groups. This is an important finding and should be considered when recommending treatment for patients with ACL injuries

Synthesis and characterization of atomically-thin graphite films on a silicon carbide substrate

This paper reports the synthesis and detailed characterization of graphite thin films produced by thermal decomposition of the (0001) face of a 6H-SiC wafer, demonstrating the successful growth of single crystalline films down to approximately one graphene layer. The growth and characterization were carried out in ultrahigh vacuum (UHV) conditions. The growth process and sample quality were monitored by low-energy electron diffraction, and the thickness of the sample was determined by core level x-ray photoelectron spectroscopy. High-resolution angle-resolved photoemission spectroscopy shows constant energy map patterns, which are very sharp and fully momentum-resolved, but nonetheless not resolution limited. We discuss the implications of this observation in connection with scanning electron microscopy data, as well as with previous studies

Molecular and Biological Characterization of the Murine Leukotriene B4 Receptor Expressed on Eosinophils

The movement of leukocytes into tissues is regulated by the local production of chemical mediators collectively referred to as chemoattractants. Although chemoattractants constitute a diverse array of molecules, including proteins, peptides, and lipids, they all appear to signal leukocytes through a related family of seven transmembrane–spanning G protein–coupled receptors. The eosinophil is a potent proinflammatory cell that is attracted into tissues during allergic inflammation, parasitic infection, and certain malignancies. Since the molecular mechanisms controlling eosinophil recruitment are incompletely understood, we performed a degenerate polymerase chain reaction on cDNA isolated from murine eosinophils to identify novel chemoattractant receptors. We report the isolation of a cDNA that encodes a 351–amino acid glycoprotein that is 78% identical to a human gene that has been reported to be a purinoceptor (P2Y7) and a leukotriene B4 (LTB4) receptor (BLTR). Chinese hamster ovary (CHO) cells transfected with this cDNA specifically bound [3H]LTB4 with a dissociation constant of 0.6 ± 0.1 nM. Furthermore, LTB4 induced a dose-dependent intracellular calcium flux in transfected CHO cells. In contrast, [35S]dATP did not specifically bind to these transfectants. This mRNA was expressed at high levels in interleukin 5–exposed eosinophils, elicited peritoneal macrophages and neutrophils, and to a lesser extent interferon γ stimulated macrophages. Low levels of expression were detected in the lung, lymph node, and spleen of unchallenged mice. Western blot analysis detected the mBLTR protein in murine eosinophils and alveolar macrophages as well as human eosinophils. In addition, elevated levels of mBLTR mRNA were found in the lungs of mice in a murine model of allergic pulmonary inflammation in a time course consistent with the influx of eosinophils. Our findings indicate that this murine receptor is an LTB4 receptor that is highly expressed on activated leukocytes, including eosinophils, and may play an important role in mediating eosinophil recruitment into inflammatory foci

GPR30 Deficiency Causes Increased Bone Mass, Mineralization, and Growth Plate Proliferative Activity in Male Mice

Estrogen regulation of the male skeleton was first clearly demonstrated in patients with aromatase deficiency or a mutation in the ERα gene. Estrogen action on the skeleton is thought to occur mainly through the action of the nuclear receptors ERα and ERβ. Recently, in vitro studies have shown that the G protein–coupled receptor GPR30 is a functional estrogen receptor (ER). GPR30-deficient mouse models have been generated to study the in vivo function of this protein; however, its in vivo role in the male skeleton remains underexplored. We have characterized size, body composition, and bone mass in adult male Gpr30 knockout (KO) mice and their wild-type (WT) littermates. Gpr30 KO mice weighed more and had greater nasal-anal length (p < .001). Both lean mass and percent body fat were increased in the KO mice. Femur length was greater in Gpr30 KO mice, as was whole-body, spine, and femoral areal bone mineral density (p < .01). Gpr30 KO mice showed increased trabecular bone volume (p < .01) and cortical thickness (p < .001). Mineralized surface was increased in Gpr30 KO mice (p < .05). Bromodeoxyuridine (BrdU) labeling showed greater proliferation in the growth plate of Gpr30 KO mice (p < .05). Under osteogenic culture conditions, Gpr30 KO femoral bone marrow cells produced fewer alkaline phosphatase–positive colonies in early differentiating osteoblast cultures but showed increased mineralized nodule deposition in mature osteoblast cultures. Serum insulin-like growth factor 1 (IGF-1) levels were not different. These data suggest that in male mice, GPR30 action contributes to regulation of bone mass, size, and microarchitecture by a mechanism that does not require changes in circulating IGF-1. © 2011 American Society for Bone and Mineral Research

Mechanisms of Vasorelaxation

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72633/1/j.1527-3466.1992.tb00249.x.pd

Leukotrienes inhibit early stages of HIV-1 infection in monocyte-derived microglia-like cells

<p>Abstract</p> <p>Background</p> <p>Microglia are one of the main cell types to be productively infected by HIV-1 in the central nervous system (CNS). Leukotriene B₄(LTB₄) and cysteinyl-leukotrienes such as LTC₄are some of the proinflammatory molecules produced in infected individuals that contribute to neuroinflammation. We therefore sought to investigate the role of leukotrienes (LTs) in HIV-1 infection of microglial cells.</p> <p>Methods</p> <p>To evaluate the role of LTs on HIV-1 infection in the CNS, monocyte-derived microglial-like cells (MDMis) were utilized in this study. Leukotriene-treated MDMis were infected with either fully replicative brain-derived HIV-1 isolates (YU2) or R5-tropic luciferase-encoding particles in order to assess viral production and expression. The efficacy of various steps of the replication cycle was evaluated by means of p24 quantification by ELISA, luciferase activity determination and quantitative real-time polymerase chain reaction (RT-PCR).</p> <p>Results</p> <p>We report in this study that virus replication is reduced upon treatment of MDMis with LTB₄and LTC₄. Additional experiments indicate that these proinflammatory molecules alter the pH-independent entry and early post-fusion events of the viral life cycle. Indeed, LT treatment induced a diminution in integrated proviral DNA while reverse-transcribed viral products remained unaffected. Furthermore, decreased C-C chemokine receptor type 5 (CCR5) surface expression was observed in LT-treated MDMis. Finally, the effect of LTs on HIV-1 infection in MDMis appears to be mediated partly via a signal transduction pathway involving protein kinase C.</p> <p>Conclusions</p> <p>These data show for the first time that LTs influence microglial cell infection by HIV-1, and may be a factor in the control of viral load in the CNS.</p