120 research outputs found
[18F]Tosyl fluoride as a versatile [18F]fluoride source for the preparation of 18F-labeled radiopharmaceuticals
Positron emission tomography (PET) is an in vivo imaging technology that utilizes positron-emitting radioisotope-labeled compounds as PET radiotracers that are commonly used in clinic and in various research areas, including oncology, cardiology, and neurology. Fluorine-18 is the most widely used PET-radionuclide and commonly produced by proton bombardment o
Role of apparent diffusion coefficients with diffusion-weighted magnetic resonance imaging in differentiating between benign and malignant bone tumors
The Rice Diacylglycerol Kinase Family: Functional Analysis Using Transient RNA Interference
Diacylglycerol kinase (DGK) is a pivotal enzyme that phosphorylates diacylglycerol (DAG) to form phosphatidic acid (PA). The production of PA from phospholipase D (PLD) and the coupled phospholipase C/DGK route is an important signaling process in animal and plant cells. In this study, we report a genomic analysis of eight putative rice DGKs encoded by a gene family (OsDGKs) grouped into three clusters. To further investigate the functions of the OsDGKs, a double-stranded RNA (dsRNA)-induced RNA silencing method was established. Introduction of in vitro-synthesized dsRNAs corresponding to a unique or conserved region of OsDGKs into rice protoplasts abolished or diminished the expression of individual or multiple OsDGK genes. Suppressing the expression of OsDGKs resulted in a distinct depletion of the transcripts of the defense gene OsNPR1 and the salt-responsive gene OsCIPK15. Our primary results suggest that OsDGKs are involved in the signaling of stress responses
Sigma-2 ligands induce tumour cell death by multiple signalling pathways
BACKGROUND: The sigma-2 receptor has been identified as a biomarker of proliferating cells in solid tumours. In the present study, we studied the mechanisms of sigma-2 ligand-induced cell death in the mouse breast cancer cell line EMT-6 and the human melanoma cell line MDA-MB-435. METHODS: EMT-6 and MDA-MB-435 cells were treated with sigma-2 ligands. The modulation of multiple signaling pathways of cell death was evaluated. RESULTS: Three sigma-2 ligands (WC-26, SV119 and RHM-138) induced DNA fragmentation, caspase-3 activation and PARP-1 cleavage. The caspase inhibitor Z-VAD-FMK partially blocked DNA fragmentation and cytotoxicity caused by these compounds. These data suggest that sigma-2 ligand-induced apoptosis and caspase activation are partially responsible for the cell death. WC-26 and siramesine induced formation of vacuoles in the cells. WC-26, SV119, RHM-138 and siramesine increased the synthesis and processing of microtubule-associated protein light chain 3, an autophagosome marker, and decreased the expression levels of the downstream effectors of mammalian target of rapamycin (mTOR), p70S6K and 4EBP1, suggesting that sigma-2 ligands induce autophagy, probably by inhibition of the mTOR pathway. All four sigma-2 ligands decreased the expression of cyclin D1 in a time-dependent manner. In addition, WC-26 and SV119 mainly decreased cyclin B1, E2 and phosphorylation of retinoblastoma protein (pRb); RHM-138 mainly decreased cyclin E2; and 10 μ siramesine mainly decreased cyclin B1 and pRb. These data suggest that sigma-2 ligands also impair cell-cycle progression in multiple phases of the cell cycle. CONCLUSION: Sigma-2 ligands induce cell death by multiple signalling pathways
PET imaging of in vivo caspase-3/7 activity following myocardial ischemia-reperfusion injury with the radiolabeled isatin sulfonamide analogue [(18)F]WC-4-116
The utility of [(18)F]WC-4-116, a PET tracer for imaging caspase-3 activation, was evaluated in an animal model of myocardial apoptosis. [(18)F]WC-4-116 was injected into rats at 3 hours after a 30 min period of ischemia induced by temporary occlusion of the left anterior descending coronary artery in Sprague-Dawley rats. [(18)F]WC-4-116 uptake was quantified by 1) autoradiography, 2) microPET imaging studies, and 3) post-PET biodistribution studies. MicroPET imaging also assessed uptake of the non-caspase-3-targeted tracer [(18)F]ICMT-18 at 3 hours postischemia. Enzyme assays and Western blotting assessed caspase-3 activation in both at-risk and not-at-risk regions. Caspase-3 enzyme activity increased in the at-risk but not in the not-at-risk myocardium. Quantitative autoradiographic analysis of [(18)F]WC-4-116 demonstrated nearly 2-fold higher uptake in the ischemia-reperfusion (IR) versus sham animals. [(18)F]WC-4-116 microPET imaging studies demonstrated that the IR animals was similarly elevated in relation to sham. [(18)F]ICMT-18 uptake did not increase in at-risk myocardium despite evidence of caspase-3 activation. Biodistribution studies with [(18)F]WC-4-116 confirmed the microPET findings. These data indicate that the caspase-3-PET tracer [(18)F]WC-4-116 can noninvasively image in vivo caspase activity during myocardial apoptosis and may be useful for clinical imaging in humans
Resorufin analogs preferentially bind cerebrovascular amyloid: potential use as imaging ligands for cerebral amyloid angiopathy
<p>Abstract</p> <p>Background</p> <p>Cerebral amyloid angiopathy (CAA) is characterized by deposition of fibrillar amyloid β (Aβ) within cerebral vessels. It is commonly seen in the elderly and almost universally present in patients with Alzheimer's Disease (AD). In both patient populations, CAA is an independent risk factor for lobar hemorrhage, ischemic stroke, and dementia. To date, definitive diagnosis of CAA requires obtaining pathological tissues via brain biopsy (which is rarely clinically indicated) or at autopsy. Though amyloid tracers labeled with positron-emitting radioligands such as [<sup>11</sup>C]PIB have shown promise for non-invasive amyloid imaging in AD patients, to date they have been unable to clarify whether the observed amyloid load represents neuritic plaques versus CAA due in large part to the low resolution of PET imaging and the almost equal affinity of these tracers for both vascular and parenchymal amyloid. Therefore, the development of a precise and specific non-invasive technique for diagnosing CAA in live patients is desired.</p> <p>Results</p> <p>We found that the phenoxazine derivative resorufin preferentially bound cerebrovascular amyloid deposits over neuritic plaques in the aged Tg2576 transgenic mouse model of AD/CAA, whereas the congophilic amyloid dye methoxy-X34 bound both cerebrovascular amyloid deposits and neuritic plaques. Similarly, resorufin-positive staining was predominantly noted in fibrillar Aβ-laden vessels in postmortem AD brain tissues. Fluorescent labeling and multi-photon microscopy further revealed that both resorufin- and methoxy-X34-positive staining is colocalized to the vascular smooth muscle (VSMC) layer of vessel segments that have severe disruption of VSMC arrangement, a characteristic feature of CAA. Resorufin also selectively visualized vascular amyloid deposits in live Tg2576 mice when administered topically, though not systemically. Resorufin derivatives with chemical modification at the 7-OH position of resorufin also displayed a marked preferential binding affinity for CAA, but with enhanced lipid solubility that indicates their use as a non-invasive imaging tracer for CAA is feasible.</p> <p>Conclusions</p> <p>To our knowledge, resorufin analogs are the fist class of amyloid dye that can discriminate between cerebrovascular and neuritic forms of amyloid. This unique binding selectivity suggests that this class of dye has great potential as a CAA-specific amyloid tracer that will permit non-invasive detection and quantification of CAA in live patients.</p
Radiolabeled 6-(2, 3-dichlorophenyl)-N4-methylpyrimidine-2, 4-diamine (TH287): A potential radiotracer for measuring and imaging MTH1
MTH1 (MutT homolog 1) or NUDT1 (Nudix Hydrolase 1), also known as oxidized purine nucleoside triphosphatase, has potential as a biomarker for monitoring cancer progression and quantifying target engagement for relevant therapies. In this study, we validate one MTH1 inhibitor TH287 as a PET MTH1 radiotracer. TH287 was radiolabeled with tritium and the binding of
Identification of the PGRMC1 protein complex as the putative sigma-2 receptor binding site
The sigma-2 receptor, whose gene remains to be cloned, has been validated as a biomarker for tumor cell proliferation. Here we report the use of a novel photoaffinity probe, WC-21, to identify the sigma-2 receptor binding site. WC-21, a sigma-2 ligand containing both a photoactive moiety azide and a fluorescein isothiocyanate group, irreversibly labels sigma-2 receptors in rat liver; the membrane-bound protein was then identified as PGRMC1 (progesterone receptor membrane component-1). Immunocytochemistry reveals that both PGRMC1 and SW120, a fluorescent sigma-2 receptor ligand, colocalizes with molecular markers of the endoplasmic reticulum and mitochondria in HeLa cells. Overexpression and knockdown of the PGRMC1 protein results in an increase and a decrease in binding of a sigma-2 selective radioligand, respectively. Th
CoNIC Challenge: Pushing the Frontiers of Nuclear Detection, Segmentation, Classification and Counting
Nuclear detection, segmentation and morphometric profiling are essential in
helping us further understand the relationship between histology and patient
outcome. To drive innovation in this area, we setup a community-wide challenge
using the largest available dataset of its kind to assess nuclear segmentation
and cellular composition. Our challenge, named CoNIC, stimulated the
development of reproducible algorithms for cellular recognition with real-time
result inspection on public leaderboards. We conducted an extensive
post-challenge analysis based on the top-performing models using 1,658
whole-slide images of colon tissue. With around 700 million detected nuclei per
model, associated features were used for dysplasia grading and survival
analysis, where we demonstrated that the challenge's improvement over the
previous state-of-the-art led to significant boosts in downstream performance.
Our findings also suggest that eosinophils and neutrophils play an important
role in the tumour microevironment. We release challenge models and WSI-level
results to foster the development of further methods for biomarker discovery
Shoulder girdle muscle activity and fatigue during automobile chassis repair
Objectives
The objective of this study was to assess the postures that were commonly used in automobile chassis repair operations, and to evaluate shoulder girdle muscle fatigue for different combinations of the weight of hand-tools.
Material and Methods
Two right muscles, including upper trapezius (UT) and middle deltoid (MD), were selected. Surface electromyography (SEMG) and a perceived level of discomfort (PLD) were used to assess the degree of shoulder girdle fatigue. Fifteen healthy young male subjects from the Northwestern Polytechnical University participated in the test. The test consisted of assuming 4 different postures and maintaining each of them for 60 s. The 4 postures varied in terms of dumbbell weights, standing for the hand-tools weight: W1 was 0.48 kg and W2 was 0.75 kg; the 4 shoulder postures were shoulder flexions of 150°, 120°, 90°, and 60°, combined with an included elbow angle of 180°, 150°, 120° and 90°, respectively. The experimental sequences were randomly selected. The signals of SEMG and the values of PLD in the shoulder girdle were recorded in 60 s. All subjects completed the whole test. The repeated measure analysis of variance (ANOVA) was performed to ascertain differences between dumbbell weight (0.48 kg and 0.75 kg) and shoulder postures (150°/180°, 120°/150°, 90°/120° and 60°/90°). The Friedman test was utilized to determine the significant differences for UT(PLD) and MD(PLD) on shoulder postures. Spearman’s correlation was used to analyze the relationship between the subjective and objective measurements.
Results
Significant correlational relationships existed between the UT percentage of the maximal voluntary electrical activation (%MVE) and UT(PLD) (r = 0.459, p < 0.01), between MD(%MVE) and MD(PLD) (r = 0.821, p < 0.01). The results showed that SEMG and PLD of the 4 postures under analysis differed significantly (p < 0.05).
Conclusions
It was indicated that posture T4 (shoulder forward flexion 60° and included elbow angle 90°) resulted in the lowest fatigue, both in terms of the objective measure and the subjective perception, which meant that this posture was more ergonomic. Int J Occup Med Environ Health. 2019;32(4):537–5
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