A new design and rationale for 3D orthogonally oversampled k‐space trajectories

A novel center‐out 3D trajectory for sampling magnetic resonance data is presented. The trajectory set is based on a single Fermat spiral waveform, which is substantially undersampled in the center of k‐space. Multiple trajectories are combined in a “stacked cone” configuration to give very uniform sampling throughout a “hub,” which is very efficient in terms of gradient performance and uniform trajectory spacing. The fermat looped, orthogonally encoded trajectories (FLORET) design produces less gradient‐efficient trajectories near the poles, so multiple orthogonal hub designs are shown. These multihub designs oversample k‐space twice with orthogonal trajectories, which gives unique properties but also doubles the minimum scan time for critical sampling of k‐space. The trajectory is shown to be much more efficient than the conventional stack of cones trajectory, and has nearly the same signal‐to‐noise ratio efficiency (but twice the minimum scan time) as a stack of spirals trajectory. As a center‐out trajectory, it provides a shorter minimum echo time than stack of spirals, and its spherical k‐space coverage can dramatically reduce Gibbs ringing. Magn Reson Med, 2011. © 2011 Wiley Periodicals, Inc.     

Dietary factors that promote or retard inflammation

Inflammation plays a pivotal role in all stages of atherosclerosis. Cardiovascular risk factors and metabolic syndrome are typified by low-grade inflammation. Intervention trials convincingly demonstrate that weight loss reduces biomarkers of inflammation, such as C-reactive protein (CRP) and interleukin (IL)-6. Limited studies have shown that certain dietary factors; oleic acid, alpha-linolenic acid, and antioxidants RRR-alpha-alpha tocopherol, reduce biomarkers of inflammation. Most of the studies with fish oil supplementation have shown null effects, and conflicting results have been reported with saturated and trans fatty acids, cholesterol, and soy intake. Much further research is needed to define the role of individual dietary factors on the biomarkers of inflammation and the mechanism of the anti-inflammatory effects of weight loss.     

C-reactive protein stimulates superoxide anion release and tissue factor activity in vivo

C-reactive protein (CRP), the prototypic marker of inflammation, is a cardiovascular risk marker and recent in vitro studies suggest that it may promote atherogenesis. CRP promotes oxidative stress in vitro and induces tissue factor (TF) release. However, there is a paucity of data examining the effects of CRP on oxidative stress and tissue factor procoagulant activity (PCA) in vivo. Thus, we tested the effects of CRP administration on superoxide anion release and tissue factor activity and examined mechanistic pathways using a rat sterile air pouch model. Intraperitoneal administration of CRP (20mg/kg body weight) compared to human serum albumin (HuSA) increased superoxide anion release and tissue factor activity from peritoneal macrophages in vivo (p<0.01). This was confirmed using intrapouch administration of CRP (25mug/mL) compared to HuSA. Pretreatment with reactive oxygen species (ROS) scavengers or protein kinase C (PKC) inhibitor significantly abrogated CRP-induced superoxide anion release and tissue factor activity. Pretreatment with extracellular signal-regulated kinase (ERK) and Jun N-terminal kinase (JNK) inhibitors, but not p38 mitogen-activated protein kinase (p38MAPK) significantly decreased CRP-induced superoxide anion release from macrophages in vivo. CRP-induced tissue factor activity in vivo was abrogated by pretreatment with inhibitors to p38MAPK, JNK and NFkappab (nuclear factor-kappab), but not ERK. Antibodies to Fc gamma receptors, CD32 and CD64 resulted in significant reduction in CRP-induced superoxide and tissue factor activity in vivo. Thus, CRP appears to induce oxidative stress in vivo by stimulating NADPH oxidase via PKC, ERK and JNK phosphorylation, and induces tissue factor PCA in vivo via upregulation of PKC, p38MAPK, JNK, ROS and NFkappab. CRP-induced ROS appears to precede tissue factor release. These effects are abrogated by blocking Fc gamma receptors, CD32 and CD64. This in vivo demonstration provides further evidence for a role for CRP in atherothrombosis.     

Increased levels of ligands of Toll-like receptors 2 and 4 in type 1 diabetes

Aims/hypothesis  Type 1 diabetes is a proinflammatory state characterised by increased levels of circulating biomarkers of inflammation and monocyte activity. We have shown increased Toll-like receptor 2 (TLR2) and TLR4 expression and signalling in monocytes from type 1 diabetic patients. Several endogenous ligands of TLR2 and TLR4 have been identified; however, there is a paucity of data on levels of these endogenous ligands in diabetes. Thus, the aim of this study was to examine circulating levels of exogenous/endogenous ligands of TLR2 and TLR4 in type 1 diabetic patients and to compare these with the levels in matched healthy controls. Methods  Healthy controls (n = 37) and type 1 diabetic patients (n = 34) were recruited, and a fasting blood sample was obtained. Circulating levels of endotoxin, heat-shock protein 60 (Hsp60), high-mobility group box 1 (HMGB1) and growth arrest-specific 6 (GAS6) proteins were assessed by ELISA, and TLR2 and TLR4 expression was determined by flow cytometry. Results  Levels of the classical TLR4 ligand, endotoxin, were significantly elevated in type 1 diabetic patients compared with those in matched controls. Hsp60 and HMGB1 concentrations were also significantly increased in the patients (p < 0.01 and p < 0.001, respectively). No significant differences were observed in GAS6. Conclusions/interpretation  We report the novel observation that levels of ligands of TLR2 and TLR4 are significantly elevated in type 1 diabetes, and this, in concert with hyperglycaemia, accounts for the increase in TLR2 and TLR4 activity, underscoring the proinflammatory state of type 1 diabetes.     

Determination of residual epichlorohydrin in sevelamer hydrochloride by static headspace gas chromatography with flame ionization detection

A sensitive static headspace gas chromatographic method was developed and validated for the determination of residual epichlorohydrin (ECH) in sevelamer hydrochloride (SVH) drug substance. This method utilized a Phenomenex Zebron ZB-WAX GC column, helium as carrier gas with flame ionization detection. The critical experimental parameters, such as, headspace vial incubation time and incubation temperature were studied and optimized. The method was validated as per United States Pharmacopoeia (USP) and International Conference on Harmonization (ICH) guidelines in terms of detection limit (DL), quantitation limit (QL), linearity, precision, accuracy, specificity and robustness. A linear range from 0.30 to 10 ?g/mL was obtained with the coefficient of determination (r(2)) 0.999. The DL and QL of ECH were 0.09 ?g/mL and 0.30 ?g/mL, respectively. The recovery obtained for ECH was between 91.7 and 96.6%. Also, the specificity of the method was proved through gas chromatography mass spectrometry (GC-MS). This method was applied successfully to determine the content of residual ECH in SVH bulk drug.     

HLA-DRβ1*04 typing by simple in-house PCR-SSP technique for rheumatoid arthritis patients

A strong association between rheumatoid arthritis (RA) and human leukocyte antigen (HLA) has been observed in many different populations and that accounts for approximately one-third of the genetic component of RA susceptibility. The greatest effect comes from DRβ1 gene where the strongest association has been found with DRβ1*04 (DR4) allele. As serology has some disadvantages over polymerase chain reaction (PCR)-based techniques and commercially available PCR-based kits are expensive, this study was aimed to standardize simple in-house PCR-SSP technique. Accuracy of this test was further checked with standard PCR-SSOP (RLS) results. The frequency HLA-DRβ1*04 was significantly increased among RA patients when compared with normal controls. In this study, a very simple, convenient and more cost-effective in-house PCR-SSP technique was standardized for HLA-DRβ1*04 typing that is helpful to RA diagnosis in developing countries like India, which can be used as a good screening test.