186 research outputs found
Hybrid SPECT/CT for the assessment of a painful hip after uncemented total hip arthroplasty
Background The diagnosis of hip pain after total hip replacement (THR)
represents a highly challenging question that is of increasing concern to
orthopedic surgeons. This retrospective study assesses bone scintigraphy with
Hybrid SPECT/CT for the diagnosis of painful THR in a selected cohort of
patients. Methods Bone SPECT/CT datasets of 23 patients (mean age 68.9 years)
with a painful hip after THR were evaluated. Selection of the patients
required an inconclusive radiograph, normal serum levels of inflammatory
parameters (CRP and ESR) or a negative aspiration of the hip joint prior to
the examination. The standard of reference was established by an
interdisciplinary adjudication-panel using all imaging data and clinical
follow-up data (>12 month). Pathological and physiological uptake patterns
were defined and applied. Results The cause of pain in this study group could
be determined in 18 out of 23 cases. Reasons were aseptic loosening (n = 5),
spine-related (n = 5), heterotopic ossification (n = 5), neuronal (n = 1),
septic loosening (n = 1) and periprosthetic stress fracture (n = 1). In (n =
5) cases the cause of hip pain could not be identified. SPECT/CT imaging
correctly identified the cause of pain in (n = 13) cases, in which the
integrated CT-information led to the correct diagnosis in (n = 4) cases,
mainly through superior anatomic correlation. Loosening was correctly assessed
in all cases with a definite diagnosis. Conclusions SPECT/CT of THA reliably
detects or rules out loosening and provides valuable information about
heterotopic ossifications. Furthermore differential diagnoses may be detected
with a whole-body scan and mechanical or osseous failure is covered by CT-
imaging. SPECT/CT holds great potential for imaging-based assessment of
painful prostheses
Surfactant phosphatidylcholine half-life and pool size measurements in premature baboons developing bronchopulmonary dysplasia
Because minimal information is available about surfactant metabolism in
bronchopulmonary dysplasia, we measured half-lives and pool sizes of
surfactant phosphatidylcholine in very preterm baboons recovering from
respiratory distress syndrome and developing bronchopulmonary dysplasia,
using stable isotopes, radioactive isotopes, and direct pool size
measurements. Eight ventilated premature baboons received (2)H-DPPC
(dipalmitoyl phosphatidylcholine) on d 5 of life, and radioactive
(14)C-DPPC with a treatment dose of surfactant on d 8. After 14 d, lung
pool sizes of saturated phosphatidylcholine were measured. Half-life of
(2)H-DPPC (d 5) in tracheal aspirates was 28 +/- 4 h (mean +/- SEM).
Half-life of radioactive DPPC (d 8) was 35 +/- 4 h. Saturated
phosphatidylcholine pool size measured with stable isotopes on d 5 was 129
+/- 14 micro mol/kg, and 123 +/- 11 micro mol/kg on d 14 at autopsy.
Half-lives were comparable to those obtained at d 0 and d 6 in our
previous baboon studies. We conclude that surfactant metabolism does not
change during the early development of bronchopulmonary dysplasia, more
specifically, the metabolism of exogenous surfactant on d 8 is similar to
that on the day of birth. Surfactant pool size is low at birth, increases
after surfactant therapy, and is kept constant during the first 2 wk of
life by endogenous surfactant synthesis. Measurements with stable isotopes
are comparable to measurements with radioactive tracers and measurements
at autopsy
Comparative HPLC-MSn analysis of canine and human meibomian lipidomes: many similarities, a few differences
The aim of this study was to evaluate the lipidome of meibomian gland secretions in canines (cMGS) – a common pet and laboratory animal – and to compare it with that of human MGS (hMGS), to determine whether canines could be used as a valid experimental animal model in studies of the biochemistry and physiology of the human ocular surface in general, and of the Meibomian glands in particular. The MGS of both species were evaluated using HPLC in combination with atmospheric pressure chemical ionization ion trap mass spectrometry. The main lipid classes found in cMGS were very long chain cholesteryl esters, wax esters, (O-acyl)-omega-hydroxy fatty acids (OAHFA), and cholesteryl esters of OAHFA. The lipidomes of cMGS and hMGS were found to be qualitatively similar, which implies similar biosynthetic and biodegradation pathways in canines and humans. However, some quantitative differences between the two were observed
Endothelial Cells' Activation and Apoptosis Induced by a Subset of Antibodies against Human Cytomegalovirus: Relevance to the Pathogenesis of Atherosclerosis
Human cytomegalovirus (hCMV) is involved in the pathogenesis of atherosclerosis. We have previously shown in patients with atherosclerosis that antibodies directed against the hCMV-derived proteins US28 and UL122 are able to induce endothelial cell damage and apoptosis of non-stressed endothelial cells through cross-rection with normally expressed surface molecules. Our aim was to dissect the molecular basis of such interaction and to investigate mechanisms linking innate immunity to atherosclerosis.We analysed the gene expression profiles in endothelial cells stimulated with antibodies affinity-purified against either the UL122 or the US28 peptides using the microarray technology. Microarray results were validated by quantitative PCR and by detection of proteins in the medium. Supernatant of endothelial cells incubated with antibodies was analysed also for the presence of Heat Shock Protein (HSP)60 and was used to assess stimulation of Toll-Like Receptor-4 (TLR4). Antibodies against UL122 and US28 induced the expression of genes encoding for adhesion molecules, chemokines, growth factors and molecules involved in the apoptotis process together with other genes known to be involved in the initiation and progression of the atherosclerotic process. HSP60 was released in the medium of cells incubated with anti-US28 antibodies and was able to engage TLR4.Antibodies directed against hCMV modulate the expression of genes coding for molecules involved in activation and apoptosis of endothelial cells, processes known to play a pivotal role in the pathogenesis of atherosclerosis. Moreover, endothelial cells exposed to such antibodies express HSP60 on the cell surface and release HSP60 in the medium able to activate TLR4. These data confirm that antibodies directed against hCMV-derived proteins US28 and UL122 purified from patients with coronary artery disease induce endothelial cell damage and support the hypothesis that hCMV infection may play a crucial role in mediating the atherosclerotic process
Cholesterol Crystals Activate the NLRP3 Inflammasome in Human Macrophages: A Novel Link between Cholesterol Metabolism and Inflammation
Chronic inflammation of the arterial wall is a key element in the pathogenesis of atherosclerosis, yet the factors that trigger and sustain the inflammation remain elusive. Inflammasomes are cytoplasmic caspase-1-activating protein complexes that promote maturation and secretion of the proinflammatory cytokines interleukin(IL)-1beta and IL-18. The most intensively studied inflammasome, NLRP3 inflammasome, is activated by diverse substances, including crystalline and particulate materials. As cholesterol crystals are abundant in atherosclerotic lesions, and IL-1beta has been linked to atherogenesis, we explored the possibility that cholesterol crystals promote inflammation by activating the inflammasome pathway.Here we show that human macrophages avidly phagocytose cholesterol crystals and store the ingested cholesterol as cholesteryl esters. Importantly, cholesterol crystals induced dose-dependent secretion of mature IL-1beta from human monocytes and macrophages. The cholesterol crystal-induced secretion of IL-1beta was caspase-1-dependent, suggesting the involvement of an inflammasome-mediated pathway. Silencing of the NLRP3 receptor, the crucial component in NLRP3 inflammasome, completely abolished crystal-induced IL-1beta secretion, thus identifying NLRP3 inflammasome as the cholesterol crystal-responsive element in macrophages. The crystals were shown to induce leakage of the lysosomal protease cathepsin B into the cytoplasm and inhibition of this enzyme reduced cholesterol crystal-induced IL-1beta secretion, suggesting that NLRP3 inflammasome activation occurred via lysosomal destabilization.The cholesterol crystal-induced inflammasome activation in macrophages may represent an important link between cholesterol metabolism and inflammation in atherosclerotic lesions
Encoding optical control in LCK kinase to quantitatively investigate its activity in live cells.
LCK is a tyrosine kinase that is essential for initiating T-cell antigen receptor (TCR) signaling. A complete understanding of LCK function is constrained by a paucity of methods to quantitatively study its function within live cells. To address this limitation, we generated LCK*, in which a key active-site lysine is replaced by a photocaged equivalent, using genetic code expansion. This strategy enabled fine temporal and spatial control over kinase activity, thus allowing us to quantify phosphorylation kinetics in situ using biochemical and imaging approaches. We find that autophosphorylation of the LCK active-site loop is indispensable for its catalytic activity and that LCK can stimulate its own activation by adopting a more open conformation, which can be modulated by point mutations. We then show that CD4 and CD8, T-cell coreceptors, can enhance LCK activity, thereby helping to explain their effect in physiological TCR signaling. Our approach also provides general insights into SRC-family kinase dynamics
Expansion and Characterization of Human Melanoma Tumor-Infiltrating Lymphocytes (TILs)
Various immunotherapeutic strategies for cancer are aimed at augmenting the T cell response against tumor cells. Adoptive cell therapy (ACT), where T cells are manipulated ex vivo and subsequently re-infused in an autologous manner, has been performed using T cells from various sources. Some of the highest clinical response rates for metastatic melanoma have been reported in trials using tumor-infiltrating lymphocytes (TILs). These protocols still have room for improvement and furthermore are currently only performed at a limited number of institutions. The goal of this work was to develop TILs as a therapeutic product at our institution.TILs from 40 melanoma tissue specimens were expanded and characterized. Under optimized culture conditions, 72% of specimens yielded rapidly proliferating TILs as defined as at least one culture reaching ≥3×10(7) TILs within 4 weeks. Flow cytometric analyses showed that cultures were predominantly CD3+ T cells, with highly variable CD4+:CD8+ T cell ratios. In total, 148 independent bulk TIL cultures were assayed for tumor reactivity. Thirty-four percent (50/148) exhibited tumor reactivity based on IFN-γ production and/or cytotoxic activity. Thirteen percent (19/148) showed specific cytotoxic activity but not IFN-γ production and only 1% (2/148) showed specific IFN-γ production but not cytotoxic activity. Further expansion of TILs using a 14-day "rapid expansion protocol" (REP) is required to induce a 500- to 2000-fold expansion of TILs in order to generate sufficient numbers of cells for current ACT protocols. Thirty-eight consecutive test REPs were performed with an average 1865-fold expansion (+/- 1034-fold) after 14 days.TILs generally expanded efficiently and tumor reactivity could be detected in vitro. These preclinical data from melanoma TILs lay the groundwork for clinical trials of ACT
Increased expression of p38 mitogen- activated protein kinase is related to the acute renal lesions induced by gentamicin
Identification of genes related to high royal jelly production in the honey bee (Apis mellifera) using microarray analysis
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