Bigels-oleocolloid matrices-as probiotic protective systems in yogurt.

The probiotic yogurt market is strong because of the potential benefits that probiotics provide to the host, such as relieving lactose intolerance symptoms, easing diarrhea, and improving the immune system. However, probiotics are sensitive to processing conditions and the high acidity of yogurt can reduce survival of probiotics and limit yogurt shelf life. Here, oleocolloid technology (bigels) was used to improve the survival of probiotics during yogurt shelf life. Bigels are semisolid systems containing a polar and a non-polar phase mixed forming a material with improved properties. Probiotic bigels were prepared by mixing a soy lecithin-stearic acid oleogel emulsion and a whey protein hydrogel, followed by the incorporation of Lactobacillus acidophilus and Bifidobacterium lactis suspended in milk. Yogurt was prepared with 18% wt/wt probiotic bigels with (Swiss-style) and without (sundae-style) agitation. Probiotic viability was monitored for 6 weeks. The total counts of L. acidophilus and B. lactis entrapped in bigels were significantly higher than free bacteria in yogurt after 3 and 5 weeks, respectively, indicating that probiotics could be entrapped and their survival enhanced. Both yogurt styles showed a meant total count of 3.3 and 4.5 log CFU/g for L. acidophilus and B. lactis, respectively at the end of storage time suggesting that despite agitation of yogurt, bigel structure played a key role in protecting probiotic viability.

Longitudinal [18F]FDG and [13N]NH3 PET/CT imaging of brain and spinal cord in a canine hemisection spinal cord injury model.

To further understand the neurological changes induced by spinal cord injury (SCI) in its acute and subacute stages, we evaluated longitudinal changes in glucose and glutamate metabolism in the spinal cord and brain regions of a canine hemisection SCI model. [18F]FDG and [13N]NH3 positron-emission tomography (PET) with computed tomography (CT) was performed before SCI and at 1, 3, 7, 14, and 21 days after SCI. Spinal cord [18F]FDG uptake increased and peaked at 3 days post SCI. Similar changes were observed in the brain regions but were not statistically significant. Compared to the acute phase of SCI, [13N]NH3 uptake increased in the subacute stage and peaked at 7 days post SCI in all analyzed brain regions. But in spinal cord, no [13N]NH3 uptake was detected before SCI when the blood-spinal cord barrier (BSCB) was intact, then gradually increased when the BSCB was damaged after SCI. [13N]NH3 uptake was significantly correlated with plasma levels of the BSCB disruption marker, monocyte chemoattractant protein-1 (MCP-1). Overall, we showed that SCI induced in vivo changes in glucose uptake in both the spinal cord and the examined brain regions, and changes in glutamine synthetase activity in the latter. Moreover, our results suggest that [13N]NH3 PET may serve as a potential method for assessing BSCB permeability in vivo.

A longitudinal 18F-fluorodeoxyglucose (18F-FDG) and 18F-sodium fluoride (18F-NaF) positron emission tomography/computed tomography (PET/CT) study in apolipoprotein E (ApoE) knockout rats fed with a Western diet.

BACKGROUND: Inflammation and vascular calcification are risk factors for cardiovascular disease, but their relationship is still under investigation. This longitudinal in vivo study aimed to monitor inflammation and calcification during the formation of atherosclerotic plaques in apolipoprotein E knockout (ApoE-/-) rats by 18F-fluorodeoxyglucose (18F-FDG) and 18F-sodium fluoride (18F-NaF) positron emission tomography/computed tomography (PET/CT). METHODS: In the ApoE group, male ApoE-/- rats were fed a high-fat Western diet from 13 weeks of age, and in the normal group, male SD rats of the same age were fed a normal diet. A longitudinal PET/CT study using 18F-FDG and 18F-NaF was performed at 12, 27, and 46 weeks of age. T1-weighted magnetic resonance imaging (MRI) was used as an atlas template, and the uptake of the tracers in the cardiovascular system was analyzed based on atlas 3D geometry volumes-of-interest (VOIs). After the PET/CT study, pathological and immunohistochemical examinations were performed on the corresponding lesions. RESULTS: The body weight and plasma cholesterol levels of the ApoE-/- rats increased with time, and at each time point, significantly higher body weight and plasma cholesterol levels were observed in the ApoE-/- rats than in the normal rats. PET/CT showed that in ApoE-/- rats, the uptake of 18F-FDG was found in the aortic arch, while the uptake of 18F-NaF was found in pulmonary arteries. The uptake of the two tracers in the ApoE group increased with time. Extensive early stage of atherosclerotic plaques, with high expression of CD68 and alizarin red, were observed in pulmonary arteries. However, only a thickened intima with very high expression of hypoxia-inducible factor-1 alpha (HIF-1α) was seen in the aortic arch. CONCLUSIONS: In ApoE-/- rats fed a high-fat Western diet, early atherosclerotic lesions developed in the pulmonary arteries; however, 18F-FDG failed to accumulate in these lesions but to accumulate in the aortic arch with only neointimal hyperplasia and significantly high expression of hypoxia.

Intravenous transplantation of olfactory ensheathing cells reduces neuroinflammation after spinal cord injury via interleukin-1 receptor antagonist.

Rationale: Olfactory ensheathing cell (OEC) transplantation has emerged as a promising therapy for spinal cord injury (SCI) repair. In the present study, we explored the possible mechanisms of OECs transplantation underlying neuroinflammation modulation. Methods: Spinal cord inflammation after intravenous OEC transplantation was detected in vivo and ex vivo by translocator protein PET tracer [18F]F-DPA. To track transplanted cells, OECs were transduced with enhanced green fluorescent protein (eGFP) and HSV1-39tk using lentiviral vector and were monitored by fluorescence imaging and [18F]FHBG study. Protein microarray analysis and ELISA studies were employed to analyze differential proteins in the injured spinal cord after OEC transplantation. The anti-inflammation function of the upregulated protein was also proved by in vitro gene knocking down experiments and OECs/microglia co-culture experiment. Results: The inflammation in the spinal cord was decreased after OEC intravenous transplantation. The HSV1-39tk-eGFP-transduced OECs showed no accumulation in major organs and were found at the injury site. After OEC transplantation, in the spinal cord tissues, the interleukin-1 receptor antagonist (IL-1Ra) was highly upregulated while many chemokines, including pro-inflammatory chemokines IL-1α, IL-1ß were downregulated. In vitro studies confirmed that lipopolysaccharide (LPS) stimulus triggered OECs to secrete IL-1Ra. OECs significantly suppressed LPS-stimulated microglial activity, whereas IL-1Ra gene knockdown significantly reduced their ability to modulate microglial activity. Conclusion: The OECs that reached the lesion site were activated by the release of pro-inflammatory cytokines from activated microglia in the lesion site and secreted IL-1Ra to reduce neuroinflammation. Intravenous transplantation of OECs has high therapeutic effectiveness for the treatment of SCI via the secretion of IL-1Ra to reduce neuroinflammation.

Identification of potential oxidative stress biomarkers for spinal cord injury in erythrocytes using mass spectrometry.

Oxidative stress is a hallmark of secondary injury associated with spinal cord injury. Identifying stable and specific oxidative biomarkers is of important significance for studying spinal cord injury-associated secondary injury. Mature erythrocytes do not contain nuclei and mitochondria and cannot be transcribed and translated. Therefore, mature erythrocytes are highly sensitive to oxidative stress and may become a valuable biomarker. In the present study, we revealed the proteome dynamics of protein expression in erythrocytes of beagle dogs in the acute and subacute phases of spinal cord injury using mass spectrometry-based approaches. We found 26 proteins that were differentially expressed in the acute (0-3 days) and subacute (7-21 days) phases of spinal cord injury. Bioinformatics analysis revealed that these differentially expressed proteins were involved in glutathione metabolism, lipid metabolism, and pentose phosphate and other oxidative stress pathways. Western blot assays validated the differential expression of glutathione synthetase, transaldolase, and myeloperoxidase. This result was consistent with mass spectrometry results, suggesting that erythrocytes can be used as a novel sample source of biological markers of oxidative stress in spinal cord injury. Glutathione synthetase, transaldolase, and myeloperoxidase sourced from erythrocytes are potential biomarkers of oxidative stress after spinal cord injury. This study was approved by the Experimental Animal Centre of Ningxia Medical University, China (approval No. 2017-073) on February 13, 2017.

Graphdiyne nanosheets as a platform for accurate copper(ii) ion detection via click chemistry and fluorescence resonance energy transfer.

A novel sensing platform for sensitive detection of copper(ii) ions (Cu2+) in living cells and body fluids was developed by taking advantage of the excellent fluorescence quenching ability of graphdiyne (GDY) and the high specificity of click chemistry for the first time.

Plasma Erythropoietin, IL-17A, and IFNγ as Potential Biomarkers of Motor Function Recovery in a Canine Model of Spinal Cord Injury.

Traumatic spinal cord injury (SCI) is a devastating neurological disease for which an accurate, cost-effective prediction of motor function recovery is in pressing need. A plethora of neurochemical changes involved in the pathophysiological process of SCI may serve as a new source of biomarkers for patient outcomes. Five dogs were included in this study. We characterized the plasma cytokine profiles in acute phase (0, 1, and 3 days after SCI) and subacute phase (7, 14, and 21 days after SCI) with microarray analysis. The motor function recovery following SCI was monitored by Olby scores. The expression level of differentially expressed proteins (DEPs) was measured with enzyme-linked immunosorbent assay (ELISA). Then, correlations with the Olby scores and receiver operating characteristic curve (ROC) analysis were performed. We identified 12 DEPs including 10 pro-inflammatory and 2 anti-inflammatory cytokines during the 21-day study period. Among those, the expression levels of erythropoietin (EPO), IL-17A, and IFNγ significantly correlated with the Olby scores with R2 values of 0.870, 0.740, and 0.616, respectively. The results of the ROC analysis suggested that plasma EPO, IL-17A, and IFNγ exhibited a significant predictive power with an area under the curve (AUC) of 0.656, 0.848, and 0.800 for EPO, IL-17A, and IFNγ, respectively. Our results provide a longitudinal description of the changes in plasma cytokine expression in the acute and subacute stages of canine SCI. These data reveal novel panels of inflammation-related cytokines which have the potential to be evaluated as biomarkers for predicting motor function prognosis after SCI.

Intravenous transplantation of olfactory bulb ensheathing cells for a spinal cord hemisection injury rat model.

Cellular transplantation strategies utilizing intraspinal or intrathecal olfactory ensheathing cells (OECs) have been reported as beneficial for spinal cord injury (SCI). However, there are many disadvantages of these methods, including additional trauma to the spinal cord parenchyma and technical challenges. Therefore, we investigated the feasibility and potential benefits of intravenous transplantation of OECs in a rat hemisection SCI model. OECs derived from olfactory bulb tissue were labeled with quantum dots (QDs), and their biodistribution after intravenous transplantation was tracked using a fluorescence imaging system. Accumulation of the transplanted OECs was observed in the injured spinal cord within 10 min, peaked at seven days after cell transplantation, and decreased gradually thereafter. This time window corresponded to the blood-spinal cord barrier (BSCB) opening time, which was quantitated with the Evans blue leakage assay. Using immunohistochemistry, we examined neuronal growth (GAP-43), remyelination (MBP), and microglia (Iba-1) reactions at the lesion site. Motor function recovery was also measured using a classic open field test (Basso, Beattie and Bresnahan score). Compared with the group injected only with QDs, the rats that received OEC transplantation exhibited a prominent reduction in inflammatory responses, increased neurogenesis and remyelination, and significant improvement in motor function. We suggest that intravenous injection could also be an effective method for delivering OECs and improving functional outcomes after SCI. Moreover, the time course of BSCB disruption provides a clinically relevant therapeutic window for cell-based intervention.

Noninvasive Assessment of Carotid Plaques Calcification by 18F-Sodium Fluoride Accumulation: Correlation with Pathology.

BACKGROUD: Vascular calcification is currently recognized as an important pathobiological process in atherosclerosis, but the mechanism remains elusive. Given the similarities in vascular calcification and bone formation, 18F-sodium fluoride (18F-NaF) is now considered a novel marker of vascular calcification. This study aimed to correlate 18F-NaF accumulation with the histological characterization of vascular calcification in carotid plaques. METHODS: A total of 8 patients who were undergoing carotid endarterectomy (CEA) for carotid artery stenosis were recruited. Before CEA, 18F-NaF positron emission tomography and computed tomography (PET-CT) studies were conducted. 18F-NaF uptake was measured by the maximum standardized uptake value and the target-to-background ratio. The Hounsfield unit (HU) value was also measured. Postoperative carotid plaques were investigated by hematoxylin and eosin staining, alizarin red staining, and immunohistochemistry (alpha-smooth muscle actin and CD68). RESULTS: 18F-NaF uptake was observed in the bilateral carotid bifurcation of all patients. Compared with the pathology results, there was a significant correlation between tracer activity in the carotid plaques and the calcification in the corresponding histological sections (integrated optical density [IOD]: r = .781, P = .022; positive area: r = .765, P = .027). A negative correlation was observed between 18F-NaF uptake and smooth muscle cell staining (IOD: r = -.710, P = .049). 18F-NaF uptake did not correlate with carotid artery stenosis, HU value, or inflammation. CONCLUSIONS: 18F-NaF PET-CT is a noninvasive imaging method for the assessment of calcification in human carotid atherosclerotic plaques and a promising approach to studying calcification in atherosclerotic lesions.

99m Tc-labeled rHuEpo for imaging of the erythropoietin receptor in tumors.

To analyze erythropoietin receptor (EpoR) status in tumors, recombinant human erythropoietin (rHuEpo) was labeled with 99m Tc by 99m Tc-centered 1-pot synthesis, resulting in high radiochemical purity, stability, and biological activity. Both in vitro cell culture experiments and biodistribution studies of normal rats demonstrated successful EpoR targeting. The biodistribution of labeled rHuEpo in a NCI-H1975 xenograft model showed tumor accumulation (tumor-to-muscle ratio, 4.27 ± 1.77), confirming the expression of active EpoR in tumors. Thus, as a novel single positron emission computerized tomography tracer for the imaging of EpoR expression in vivo, 99m Tc-rHuEpo is effective for exploring the role of EpoR in cancer growth, metastasis and angiogenesis.

Primary Pulmonary Artery Sarcoma on Dual-Time Point FDG PET/CT Imaging.

A 59-year-old man presented cough, chest pain, and shortness of breath for 2 weeks and fever for 4 days. A contrast chest CT revealed a large right pulmonary artery filling defect, suggestive of pulmonary embolism that failed to respond to anticoagulation therapy. FDG PET/CT was performed to evaluate possible malignancy, which revealed intense activity in the right main pulmonary artery without any extrathoracic abnormality. The ratio of the SUVmax of this lesion to the liver was significantly increased in the delayed PET images. The pathological examination demonstrated primary pulmonary artery sarcoma.

Cationic Nanoparticles Assembled from Natural-Based Steroid Lipid for Improved Intracellular Transport of siRNA and pDNA.

Developing new functional biomaterials from biocompatible natural-based resources for gene/drug delivery has attracted increasing attention in recent years. In this work, we prepared a series of cationic nanoparticles (Diosarg-DOPE NPs) by assembly of a natural steroid diosgenin-based cationic lipid (Diosarg) with commercially-available helper lipid 1,2-dioleoyl-sn-glycero-3-phosphorethanolamine (DOPE). These cationic Diosarg-DOPE NPs were able to efficiently bind siRNA and plasmid DNA (pDNA) via electrostatic interactions to form stable, nano-sized cationic lipid nanoparticles instead of lamellar vesicles in aqueous solution. The average particle size, zeta potentials and morphologies of the siRNA and pDNA complexes of the Diosarg-DOPE NPs were examined. The in vitro cytotoxicity of NPs depends on the dose and assembly ratio of the Diosarg and DOPE. Notably, the intracellular transportation efficacy of the exogenesis siRNA and pDNA could be greatly improved by using the Diosarg-DOPE NPs as the cargoes in H1299 cell line. The results demonstrated that the self-assembled Diosarg-DOPE NPs could achieve much higher intracellular transport efficiency for siRNA or pDNA than the cationic lipid Diosarg, indicating that the synergetic effect of different functional lipid components may benefit the development of high efficiency nano-scaled gene carriers. Moreover, it could be noted that the traditional "lysosome localization" involved in the intracellular trafficking of the Diosarg and Diosarg-DOPE NPs, indicating the co-assembly of helper lipid DOPE, might not significantly affect the intracellular localization features of the cationic lipids.

Ratiometric fluorescence sensor based on a pyrene derivative and quantification detection of heparin in aqueous solution and serum.

A ratiometric fluorescence sensor based on pyrene was designed for selective detection of heparin in HEPES (N-(2-hydroxyethyl)piperazine-N'-ethanesulfonic acid) buffer and serum sample. Pyrene and long-chain alkanes were linked through bisquaternary functionality in the sensor which could interact with heparin via supramolecular assembly. A ratiometric fluorescent signal change of the sensor can be observed because of the specific monomer-excimer conversion of pyrene which is modulated by the supramolecular self-assembly of sensor and heparin. Upon addition of heparin, the excimer emission of the sensor at 489 nm is observed and the monomer emission intensity at 395 nm decreases concomitantly. Addition of heparin derivatives with very similar structure such as chondroitin 4-sulfate or hyaluronic acid to the same sensor solution only leads to very smaller changes in intensity ratios probably because of lower charge density and more distant spatial distribution of anions (or disadvantageous spatial orientation of anions) as compared to those of heparin. The novel sensor can effectively differentiate heparin from its derivatives with relatively low background interference and wide linear response in HEPES and serum. A linear calibration curve is obtained from 0 to 3.4 µM for heparin quantification in serum.

Reversible fluorescent probe for highly selective and sensitive detection of mercapto biomolecules.

A coumarin-derived complex, Hg(2)L(2), was reported as a highly sensitive and selective probe for the detection of mercapto biomolecules in aqueous solution. The addition of Cys to a 99% aqueous solution of Hg(2)L(2) resulted in rapid and remarkable fluorescence OFF-ON (emission at 525 nm) due to the ligand-exchange reaction of Cys with L coordinated to Hg(2+). The increased fluorescence can be completely quenched by Hg(2+) and recovered again by the subsequent addition of Cys. Such a fluorescence OFF-ON circle can be repeated at least 10 times by the alterative addition of Cys and Hg(2+) to the solution of Hg(2)L(2), indicating that it can be used as a convertible and reversible probe for the detection of Cys. The interconversion of Hg(2)L(2) and L via the decomplexation/complexation by the modulation of Cys/Hg(2+) was definitely verified from their crystal structures. Other competitive amino acids without a thiol group cannot induce any fluorescence changes, implying that Hg(2)L(2) can selectively determine mercapto biomolecules. Using confocal fluorescence imaging, L/Hg(2)L(2) as a pair of reversible probes can be further applied to track and monitor the self-detoxification process of Hg(2+) ions in SYS5 cells.

A novel fluoride ion colorimetric chemosensor based on coumarin.

A novel visible colorimetric sensor (L1) with high selectivity for fluoride ion based on coumarin has been synthesized by a simple modification of our earlier report. The chemosensor L1 shows an obvious color change from yellow to blue upon addition of fluoride ion with a large red shift of 145 nm in acetonitrile, and without interference of other anions such as Cl-, Br-, I-, NO3-, H2PO4-, HSO4-, and AcO-. The investigation of 1H NMR spectrum titration indicates the proposed mechanism is that F- first establishes a hydrogen bonding interaction with L1, and then the formation of [F-H-F]- induces deprotonation.

Enzyme sensing based on a controllable oxidation reaction.

We developed a new method for glutathione reductase (GR) enzyme sensing via a metal-controlled spontaneous oxidation reaction. A new complex HgL, composed of 3-benzothiazoliny-7-N,N-diethylaminocoumarin (L) and HgCl2, was used as an example for illustration. It was found that ligand L was released from complex HgL by a ligand exchange process in the presence of GSH, which was enzymatic reduced from GSSG in the presence of NADH. Subsequently, free ligand L was spontaneously oxidated to a laser dye (coumarin-6) with both fluorescence enhancement and color change. A good linear relationship between fluorescent intensity and GSH concentrations (in the range of 10-40 µM) elicits the potential use of HgL in GSH detection. Based on this result, complex HgL was applied to identify GR from other proteins/enzymes with good selectivity and sensitivity. HgL can also determine GR in the concentration range from 0.5 to 10 U/mL with a linear relationship. Furthermore, this novel sensing method was also expected to be a useful example for the design of other biosensing systems.

A colorimetric and ratiometric fluorescent probe for thiols and its bioimaging applications.

A naphthalimide-based colorimetric fluorescent probe containing a disulfide group was designed and synthesized, which could detect the physiological level of GSH quantitatively by a ratiometric fluorescence method and was successfully applied to the imaging of thiols in living HeLa cells.

Highly selective recognition of carbenicillin via concerted interactions in 100% aqueous solution.

A fluorescent probe (QA) based on quinine bearing two quaternary ammonium groups and a long hydrophobic chain was synthesized, and it showed highly selective recognition of carbenicillin (a typical beta-lactam antibiotic) in 100% aqueous solution via concerted interactions.

Highly selective and sensitive heparin probing from supramolecular assembly of pyrene derivatives.

A new quinine derivative PQ, bearing pyrenyl as a fluorophore, was prepared to recognize heparin in aqueous solution. PQ exhibits good selectivity and sensitivity for heparin over other biological molecules. Upon binding with heparin, PQ shows a typical excimer emission peaked at 489 nm along with a weak monomer emission at 376 nm. Moreover, PQ also shows good performance to detect the heparin in serum.