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1
Ouyang, Zhiming, and Richard Isaacson. "Identification and Characterization of a Novel ABC Iron Transport System, fit, in Escherichia coli." Infection and Immunity 74, no. 12 (September 18, 2006): 6949–56. http://dx.doi.org/10.1128/iai.00866-06.
Повний текст джерелаАнотація:
ABSTRACT A putative ABC transporter, fit, with significant homology to several bacterial iron transporters was identified in Escherichia coli. The E. coli fit system consists of six genes designated fitA, -B, -C, -D, -E, and -R. Based on DNA sequence analysis, fit encodes an outer membrane protein (FitA), a periplasmic binding protein (FitE), two permease proteins (FitC and -D), an ATPase (FitB), and a hypothetical protein (FitR). Introduction of the E. coli fit system into E. coli strain K-12 increased intracellular iron content and transformed bacteria were more sensitive to streptonigrin, which suggested that fit transports iron in E. coli. Expression of fit was studied using a lacZ reporter assay. A functional, bidirectional promoter was identified in the intergenic region between genes fitA and fitB. The expression of the E. coli fit system was found to be induced by iron limitation and repressed when Fe2+ was added to minimal medium. Several fit mutants were created in E. coli using an in vitro transposon mutagenesis strategy. Mutations in fit did not affect bacterial growth in iron-restricted media. Using a growth promotion test, it was found that fit was not able to transport enterobactin, ferrichrome, transferrin, and lactoferrin in E. coli.
2
Adamski, S. W., J. J. Langone, and G. J. Grega. "Modulation of macromolecular permeability by immune-complexes and a beta-adrenoceptor stimulant." American Journal of Physiology-Heart and Circulatory Physiology 253, no. 6 (December 1, 1987): H1586—H1595. http://dx.doi.org/10.1152/ajpheart.1987.253.6.h1586.
Повний текст джерелаАнотація:
The suffused, noneverted cheek pouch of ovalbumin (OA)-immunized hamsters was employed to study changes in vascular permeability utilizing intravital light microscopy and direct measurements of plasma to suffusate tracer efflux. Suffusion of the cheek pouches of immunized animals with OA for 10 min stimulated the formation of focal venular fluorescein isothiocyanate dextran (FITC-D, 70,000 Da) leakage sites and produced increases in the plasma to suffusate FITC-D efflux resulting in marked increases in concentration of FITC-D in suffusate ([FITC-D]s). The intravenous injection of the FITC-D 15 to 60 min after the start of the OA suffusion failed to reveal the formation of vascular FITC-D leakage sites or increases in [FITC-D]s. In contrast, the intravenous injection of the tracer at the start or 10 min after the start of the OA suffusion revealed the formation of venular FITC-D leakage sites and marked increases in [FITC-D]s. Treatment with diphenhydramine or isoproterenol completely inhibited the antigen-stimulated formation of venular FITC-D leakage sites and increases in [FITC-D]s. It is concluded that antigen-antibody reactions resulting in the production of immune complexes trigger immediate increases in the plasma to suffusate FITC-D clearance via the formation of venular FITC-D leakage sites subsequent to the release of endogenous histamine. The increase in venular permeability is marked but transient, lasting approximately 10 min, and is subject to inhibition by either diphenhydramine or isoproterenol.
3
Burns, A. R., S. P. Hosford, L. A. Dunn, D. C. Walker, and J. C. Hogg. "Respiratory epithelial permeability after cigarette smoke exposure in guinea pigs." Journal of Applied Physiology 66, no. 5 (May 1, 1989): 2109–16. http://dx.doi.org/10.1152/jappl.1989.66.5.2109.
Повний текст джерелаАнотація:
The purpose of this study was to determine the pathology of cigarette smoke-increased permeability at the bronchioalveolar junction of the guinea pig. After exposure to either smoke or room air, guinea pigs were anesthetized and fluorescein isothiocyanate-dextran (FITC-D, mol wt 10,000) was aerosolized into their lungs. Blood samples taken through a carotid arterial cannula were analyzed by gel chromatography and spectrofluorometry for the presence of FITC-D. The results confirmed that, after smoke exposure, increased amounts of intact FITC-D molecules with a reported Einstein-Stokes radius of 22.2 A crossed the respiratory epithelium into the vascular space. Transmission electron-microscopic studies showed that the FITC-D diffused across damaged type I pneumocyte membranes and cytoplasm to reach the basal lamina and entered the alveolar capillaries through endothelial tight junctions. Damage to the alveolar epithelium was more frequent for the smoke-exposed animals than the room air-exposed animals (P less than 0.05). We conclude that smoke exposure damages type I cells and that inhaled FITC-D crosses the epithelial barrier at damaged type I cells of the bronchioloalveolar junctions.
4
Matsuda, T., I. Rubinstein, R. A. Robbins, S. Koyama, W. L. Joyner, and S. I. Rennard. "Role of neutrophils in endotoxin-mediated microvascular injury in hamsters." Journal of Applied Physiology 71, no. 1 (July 1, 1991): 307–13. http://dx.doi.org/10.1152/jappl.1991.71.1.307.
Повний текст джерелаАнотація:
The purpose of this study was to examine the role of circulating neutrophils in endotoxin-induced increase in microvascular permeability in vivo. Fifteen hamsters were anesthetized, and a plastic chamber was placed in each cheek pouch to observe the microvasculature. Fluorescein-labeled dextran (FITC-D, 150 kDa) was injected intravenously, and changes in leaky sites and FITC-D clearance were measured in three groups: control (saline, n = 4), endotoxin suffusion (n = 6), and endotoxin suffusion after neutropenia induction (n = 5). We found a significant increase in leaky sites and FITC-D clearance with endotoxin (45 +/- 18/cm2 and 20 +/- 6 x 10(-6) ml/min, respectively; mean +/- SD, P less than 0.05) in comparison to control (7 +/- 6/cm2 and 7 +/- 5 x 10(-6) ml/min) and endotoxin suffusion in neutropenic animals (19 +/- 11/cm2 and 12 +/- 4 x 10(-6) ml/min). There was a significant correlation between the number of leaky sites and FITC-D clearance (r = 0.91, P less than 0.01) and between the number of circulating neutrophils and FITC-D clearance (r = 0.87, P less than 0.01). We conclude that endotoxin-mediated increase in microvascular permeability in the peripheral circulation is dependent in part on circulating neutrophils.
5
Seksek, Olivier, Joachim Biwersi, and A. S. Verkman. "Translational Diffusion of Macromolecule-sized Solutes in Cytoplasm and Nucleus." Journal of Cell Biology 138, no. 1 (July 14, 1997): 131–42. http://dx.doi.org/10.1083/jcb.138.1.131.
Повний текст джерелаАнотація:
Fluorescence recovery after photobleaching (FRAP) was used to quantify the translational diffusion of microinjected FITC-dextrans and Ficolls in the cytoplasm and nucleus of MDCK epithelial cells and Swiss 3T3 fibroblasts. Absolute diffusion coefficients (D) were measured using a microsecond-resolution FRAP apparatus and solution standards. In aqueous media (viscosity 1 cP), D for the FITC-dextrans decreased from 75 to 8.4 × 10−7 cm2/s with increasing dextran size (4–2,000 kD). D in cytoplasm relative to that in water (D/Do) was 0.26 ± 0.01 (MDCK) and 0.27 ± 0.01 (fibroblasts), and independent of FITC-dextran and Ficoll size (gyration radii [RG] 40–300 Å). The fraction of mobile FITC-dextran molecules (fmob), determined by the extent of fluorescence recovery after spot photobleaching, was >0.75 for RG < 200 Å, but decreased to <0.5 for RG > 300 Å. The independence of D/Do on FITC-dextran and Ficoll size does not support the concept of solute “sieving” (size-dependent diffusion) in cytoplasm. Photobleaching measurements using different spot diameters (1.5–4 μm) gave similar D/Do, indicating that microcompartments, if present, are of submicron size. Measurements of D/Do and fmob in concentrated dextran solutions, as well as in swollen and shrunken cells, suggested that the low fmob for very large macromolecules might be related to restrictions imposed by immobile obstacles (such as microcompartments) or to anomalous diffusion (such as percolation). In nucleus, D/Do was 0.25 ± 0.02 (MDCK) and 0.27 ± 0.03 (fibroblasts), and independent of solute size (RG 40–300 Å). Our results indicate relatively free and rapid diffusion of macromolecule-sized solutes up to approximately 500 kD in cytoplasm and nucleus.
6
Kim, D., P. M. Armenante, and W. N. Duran. "Transient analysis of macromolecular transport across microvascular wall and into interstitium." American Journal of Physiology-Heart and Circulatory Physiology 265, no. 3 (September 1, 1993): H993—H999. http://dx.doi.org/10.1152/ajpheart.1993.265.3.h993.
Повний текст джерелаАнотація:
We studied the dynamics of macromolecular transport across the microvascular wall in the hamster cheek pouch using intravital microscopy and digital video-image analysis. We used fluorescein isothiocyanate-dextrans of 70,000 and 150,000 Da (FITC-Dextran 70 and 150, respectively) as tracers. We applied our mathematical model and our in vivo calibration to determine the diffusion coefficient (D) and the average fluid velocity (V) in the microvascular wall and in the interstitium from the experimental data. The value of D for FITC-Dextran 70 was 0.90 +/- 0.04 x 10(-11) cm2/s in the wall and 1.29 +/- 0.05 x 10(-8) cm2/s in the interstitium. In both regions, V was 2.05 +/- 0.05 x 10(-8) cm/s. The transport parameters for FITC-Dextran 150 were 0.27 +/- 0.02 x 10(-11) cm2/s, 0.55 +/- 0.05 x 10(-8) cm2/s, and 1.71 +/- 0.48 x 10(-8) cm/s for D in the wall and interstitium and V, respectively. The topical application of either calcium ionophore A23187 (7 x 10(-7) M) or bradykinin (5 x 10(-7) M) increased D for FITC-Dextran 70 and 150 2-fold and V 10-fold relative to their control values. We used these values to quantify the relative importance of the diffusive and convective mechanisms in the total solute flux. Molecular diffusion dominates convective transport in both the microvascular wall and the interstitial space.
7
Adamski, S. W., and G. J. Grega. "Contribution of kininase II to the waning of vascular actions of bradykinin." American Journal of Physiology-Heart and Circulatory Physiology 254, no. 6 (June 1, 1988): H1042—H1050. http://dx.doi.org/10.1152/ajpheart.1988.254.6.h1042.
Повний текст джерелаАнотація:
The mechanism(s) of the waning of the vasodilation and increase in vascular permeability during prolonged local intraarterial infusions of bradykinin (BK) was investigated in this study. Treatment with phentolamine or saralasin failed to prevent the waning of the vasodilation during the prolonged infusion of BK into forelimbs perfused at constant flow. In contrast, BK produced a sustained vasodilator response after treatment with captopril. Forelimb weight and lymph analysis were used to quantitate edema formation and to determine the duration of the increase in vascular permeability during prolonged local intra-arterial infusions of BK into forelimbs perfused at constant flow. The lymph-to-plasma ratios (L/P) for protein and FITC-Dextrans (fluorescein isothiocyanate dextrans, 70,000 Da) were determined, and clearances for protein and FITC-D were calculated. BK markedly increased fluid filtration, the protein L/P, and protein clearance resulting in edema formation. The protein L/P remained markedly elevated throughout the experimental period. The FITC-D L/P was markedly increased in the groups of animals in which the tracer was injected intravenously at the start or 8 min after the start of the prolonged BK infusion. In the groups of animals in which the tracer was injected intravenously 15-60 min after the start of the prolonged BK infusion, the FITC-D L/P failed to exceed the FITC-D L/P in control animals, although the protein L/P remained elevated. Pretreatment with both captopril and propranolol dramatically potentiated the magnitude of the increase in protein clearance, the filtration rate, and edema formation produced by BK but failed to affect the duration of the transient increase in vascular permeability.
8
Matsuda, T., C. A. Eccleston, I. Rubinstein, S. I. Rennard, and W. L. Joyner. "Antioxidants attenuate endotoxin-induced microvascular leakage of macromolecules in vivo." Journal of Applied Physiology 70, no. 4 (April 1, 1991): 1483–89. http://dx.doi.org/10.1152/jappl.1991.70.4.1483.
Повний текст джерелаАнотація:
The purpose of this study was to examine whether antioxidants attenuate endotoxin-induced microvascular hyper-permeability for macromolecules in the hamster cheek pouch. Twenty-two adult male Syrian hamsters were anesthetized, and a removable plastic chamber was placed in the cheek pouch to observe and collect suffusate from the microvasculature. Fluorescent-labeled dextran (FITC-D; mol wt 150,000) was injected intravenously, and changes in the number of microvascular leaky sites and microvascular clearance of FITC-D were measured in five groups: saline control (group 1, n = 4), endotoxin (0.1 mg/ml) suffusion for 120 min (group 2, n = 6), endotoxin plus dimethyl sulfoxide (1.0 g/kg iv; group 3, n = 4), endotoxin plus allopurinol (30 mg/kg ip; group 4, n = 4), and endotoxin plus dimethyl sulfoxide and allopurinol (group 5, n = 4). The number of leaky sites and the FITC-D clearance were significantly higher in group 2 [45 +/- 18 (SD) sites/cm2 and 20 +/- 6 X 10(-6) ml/min, respectively; P less than 0.01] than in group 1 (7 +/- 6 sites/cm2 and 7 +/- 5 X 10(-6) ml/min), group 3 (9 +/- 5 sites/cm2 and 8 +/- 2 X 10(-6) ml/min), group 4 (11 +/- 7 sites/cm2 and 9 +/- 4 X 10(-6) ml/min), and group 5 (11 +/- 6 sites/cm2 and 7 +/- 1 x 10(-6) ml/min). The leaky sites appeared predominantly in postcapillary venules. There was a positive and significant correlation between the number of leaky sites and FITC-D clearance.(ABSTRACT TRUNCATED AT 250 WORDS)
9
Vermaelen, Karim Y., Ines Carro-Muino, Bart N. Lambrecht, and Romain A. Pauwels. "Specific Migratory Dendritic Cells Rapidly Transport Antigen from the Airways to the Thoracic Lymph Nodes." Journal of Experimental Medicine 193, no. 1 (December 27, 2000): 51–60. http://dx.doi.org/10.1084/jem.193.1.51.
Повний текст джерелаАнотація:
Antigen transport from the airway mucosa to the thoracic lymph nodes (TLNs) was studied in vivo by intratracheal instillation of fluorescein isothiocyanate (FITC)-conjugated macromolecules. After instillation, FITC+ cells with stellate morphology were found deep in the TLN T cell area. Using flow cytometry, an FITC signal was exclusively detected in CD11cmed-hi/major histocompatibility complex class II (MHCII)hi cells, representing migratory airway-derived lymph node dendritic cells (AW-LNDCs). No FITC signal accumulated in lymphocytes and in a CD11chiMHCIImed DC group containing a CD8αhi subset (non–airway-derived [NAW]-LNDCs). Sorted AW-LNDCs showed long MHCIIbright cytoplasmic processes and intracytoplasmatic FITC+ granules. The fraction of FITC+ AW-LNDCs peaked after 24 h and had reached baseline by day 7. AW-LNDCs were depleted by 7 d of ganciclovir treatment in thymidine kinase transgenic mice, resulting in a strong reduction of FITC-macromolecule transport into the TLNs. Compared with intrapulmonary DCs, AW-LNDCs had a mature phenotype and upregulated levels of MHCII, B7-2, CD40, and intracellular adhesion molecule (ICAM)-1. In addition, sorted AW-LNDCs from FITC-ovalbumin (OVA)–instilled animals strongly presented OVA to OVA-TCR transgenic T cells. These results validate the unique sentinel role of airway DCs, picking up antigen in the airways and delivering it in an immunogenic form to the T cells in the TLNs.
10
Ivesic, Caroline, Stefanie Krammer, Marianne Koller-Peroutka, Aicha Laarouchi, Daniela Gruber, Ingeborg Lang, Irene K. Lichtscheidl, and Wolfram Adlassnig. "Quantification of Protein Uptake by Endocytosis in Carnivorous Nepenthales." Plants 12, no. 2 (January 11, 2023): 341. http://dx.doi.org/10.3390/plants12020341.
Повний текст джерелаАнотація:
Carnivorous plants adsorb prey-derived nutrients partly by endocytosis. This study quantifies endocytosis in Drosophyllum lusitanicum, Drosera capensis, Drosera roseana, Dionaea muscipula and Nepenthes × ventrata. Traps were exposed to 1% fluorescent-labeled albumin (FITC-BSA), and uptake was quantified repeatedly for 64 h. Formation of vesicles started after ≤1 h in adhesive traps, but only after 16 h in species with temporary stomach (D. muscipula and N. × ventrata). In general, there are similarities in the observed species, especially in the beginning stages of endocytosis. Nonetheless, further intracellular processing of endocytotic vesicles seems to be widely different between species. Endocytotic vesicle size increased significantly over time in all species except in D. capensis. Fluorescence intensity of the endocytotic vesicles increased in all species except D. muscipula. After 64 h, estimates for FITC-BSA absorption per gland ranged from 5.9 ± 6.3 ng in D. roseana to 47.8 ± 44.3 ng in N. × ventrata, demonstrating that endocytosis substantially contributes to the adsorption of prey-derived nutrients.
11
Gabler, Nicholas K., Dawn Koltes, Simone Schaumberger, G. Raj Murugesan, and Nicole Reisinger. "Diurnal heat stress reduces pig intestinal integrity and increases endotoxin translocation." Translational Animal Science 2, no. 1 (January 25, 2018): 1–10. http://dx.doi.org/10.1093/tas/txx003.
Повний текст джерелаАнотація:
Abstract Heat stress negatively affects performance and intestinal integrity of pigs. The objective of this study was to characterize the effects of diurnal heat stress (dHS) on nursery-grower pig performance, intestinal integrity, and lipopolysaccharide (LPS) translocation. Forty-eight nursery-grower gilts, individually penned, were randomly assigned to two treatments. Twenty-four pigs were then exposed to dHS for 3 d, 6 h at 38°C and 18 h at 32°C, at 40–60% humidity. The remaining pigs were maintained under thermal neutral (TN) conditions. Changes in pig rectal temperatures (Tr), respiration rates (RR), performance, and blood parameters were evaluated. Additionally, ex vivo ileum integrity was assessed with the Ussing chamber by measuring transepithelial resistance (TER), and 4 kDa fluorescein isothiocyanate (FITC)–dextran (FD4) and FITC–LPS mucosal to serosal flux. As expected, dHS increased pig Tr and RR (P < 0.05) and reduced pig performance (P < 0.05) on the 3-d period. Compared with TN, ileum TER (P = 0.04), FITC–LPS (P < 0.001), and FD4 (P = 0.011) permeability were significantly increased due to dHS. Compared with TN pigs, dHS increased serum endotoxin by 150% (P = 0.031). Altogether, 3-d dHS significantly reduced pig performance and intestinal integrity and increased blood endotoxin concentrations.
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Hjelle, J. Thomas, Barbara T. Golinska, Diane C. Waters, Kevin R. Steidley, Marcia A. Miller, David R. McCarrol, and James W. Dobbie. "Lectin Staining of Peritoneal Mesothelial Cells in Vitro." Peritoneal Dialysis International: Journal of the International Society for Peritoneal Dialysis 11, no. 4 (October 1991): 307–16. http://dx.doi.org/10.1177/089686089101100403.
Повний текст джерелаАнотація:
A survey of lectin-binding specificities present on rodent and human mesothelial cells propagated and maintained in tissue culture was made using fluorescein isothiocynate conjugated (FITC) lectins. Rodent and human cells exhibited cell-associated fluorescence following exposure to the FITC-Iectins from C. ensiformis, T. vulgaris, A. hypogaea, E. cristagalli and B. simplicifolia, but not with lectins from G. max and D. biflorus. Rodent cells were also positive for FITC-M. pomifera lectin binding. Human, but not rodent, cells were positive for FITC T. purpureas lectin binding. Exposure of rabbit mesothelial cells in vitro to FITC-Iectins that bound to the cell surface resulted in the appearance of discrete loci of putatively intracellular fluorescence. Exposure of cells to ferritin-Iabelled T. vulgaris lectin at 37°C for as little as 7.5 minutes resulted in the appearance of ferritin-size particles in intracellular vesicles. These results demonstrate 1. the presence of lectinbinding sites in and on peritoneal mesothelial cells from rodents and humans and 2. a possible role of such sites in mediating the entry of lectin-Iike endogenous molecules into the vacuolar apparatus of these cells.
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Mayhan, W. G. "Role of nitric oxide in leukotriene C4-induced increases in microvascular transport." American Journal of Physiology-Heart and Circulatory Physiology 265, no. 1 (July 1, 1993): H409—H414. http://dx.doi.org/10.1152/ajpheart.1993.265.1.h409.
Повний текст джерелаАнотація:
The goal of this study was to determine the role of nitric oxide in alterations in macromolecular transport of the hamster cheek pouch in vivo in response to leukotriene C4. We used intravital fluorescent microscopy to examine the transport of macromolecules across the hamster cheek pouch in response to leukotriene C4 before and after application of an enzymatic inhibitor of nitric oxide, NG-monomethyl-L-arginine (L-NMMA; 1.0 microM). Increases in transport of macromolecules across the hamster cheek pouch were quantitated by the formation of venular leaky sites and clearance of fluorescein isothiocyanate-dextran (FITC-dextran; mol wt = 70 K). Leukotriene C4 (1.0 and 3.0 nM) produced an increase in the number of venular leaky sites and clearance of FITC-dextran-70K. Superfusion of L-NMMA (1.0 microM) significantly decreased leukotriene C4-induced increases in venular leaky sites and clearance of FITC-dextran-70K. In addition, superfusion of LY-83583 (10 microM) significantly decreased leukotriene C4-induced increases in venular leaky sites. In contrast, superfusion of NG-monomethyl-D-arginine (D-NMMA; 1.0 microM), indomethacin (10 mg/kg iv), or diphenhydramine hydrochloride; 15–20 mg/kg iv) did not significantly alter leukotriene C4-induced increases in venular leaky sites. Thus these findings suggest that production of nitric oxide and subsequent activation of guanylate cyclase play an important role in formation of venular leaky sites and clearance of FITC-dextran-70K in response to application of leukotriene C4.
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Macatonia, S. E., S. C. Knight, A. J. Edwards, S. Griffiths, and P. Fryer. "Localization of antigen on lymph node dendritic cells after exposure to the contact sensitizer fluorescein isothiocyanate. Functional and morphological studies." Journal of Experimental Medicine 166, no. 6 (December 1, 1987): 1654–67. http://dx.doi.org/10.1084/jem.166.6.1654.
Повний текст джерелаАнотація:
We have examined the cells involved in the development of contact sensitivity to FITC in CBA mice. After skin painting with antigen, the number of dendritic cells (DC) in the draining lymph nodes increased by 30 min, was maximal at 48 h, and returned to normal by 6 d. Derivation of some DC from Langerhans' cells of the skin was indicated from the presence of Birbeck granules observed in some DC isolated 24 h after skin painting. The DC acquired FITC and by 8 h there were two populations, one highly fluorescent and the other less fluorescent. The highly fluorescent cells were present between 8 h and 3 d after sensitization, and during this period the DC were potent at initiating primary proliferative responses of normal syngeneic T lymphocytes in vitro. Between days 3 and 5 the numbers of lymphocytes in the draining lymph node increased. During this period purified T lymphocytes did not express detectable levels of antigen, but enriched B cell populations expressed antigen transiently on day 1, 2, or 3 after exposure to antigen. The results showed that, during a 3-d period after exposure to antigen, DC expressed antigen and stimulated T cell proliferation. We speculate that low amounts of FITC binding selectively to veiled cells or lymph node DC in the first hours after exposure to antigen are not immunogenic but that Langerhans' cells acquire high levels of antigen, enter the nodes, and initiate immune responses.
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Kumar, Alip, Sarbast K. Kheravii, Catherine Ionescu, Alexandra Blanchard, Reza Barekatain, Yadav S. Bajagai, and Shu-Biao Wu. "A Microencapsulated Mixture of Eugenol and Garlic Tincture Supplementation Mitigates the Effect of Necrotic Enteritis on Intestinal Integrity and Increases Goblet Cells in Broilers." Microorganisms 9, no. 7 (July 6, 2021): 1451. http://dx.doi.org/10.3390/microorganisms9071451.
Повний текст джерелаАнотація:
This study was conducted to examine the effects of a plant extract mixture, a microencapsulated product composed of eugenol and garlic tincture (PE), on intestinal health in broilers under necrotic enteritis (NE) challenge. A total of 960 d-old mixed-sex Cobb 500 chicks were randomly distributed to 48-floor pens housing 20 birds per pen. Six treatments were applied: UC, unchallenged control; CC, challenged control; PE, challenged group plus PE; AM, challenged group plus antimicrobial (AM); FAP, challenged group plus a full dose of AM with PE; HAP, challenged group plus a half dose of AM with PE in starter, grower and finisher phases. Birds in the challenged groups were inoculated with Eimeria spp. on d 9 and Clostridiumperfringens on d 14. On d 16, the CC group had increased serum fluorescein isothiocyanate dextran (FITC-d), reduced villus surface area, goblet cell number, upregulated CLDN1, JAM2 genes and reduced microbial diversity compared to the UC group (p < 0.05). Birds fed PE had reduced FITC-d, increased goblet cell number and Bifidobacterium compared to the CC group (p < 0.05). Birds fed PE had reduced CLDN5 expression in male birds, and Bacteroides spp. in female birds than CC group (p < 0.05). These findings suggest that PE supplementation mitigates the effect of NE by improving the intestinal health of birds.
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Vicuña, E. A., V. A. Kuttappan, G. Tellez, X. Hernandez-Velasco, R. Seeber-Galarza, J. D. Latorre, O. B. Faulkner, A. D. Wolfenden, B. M. Hargis, and L. R. Bielke. "Dose titration of FITC-D for optimal measurement of enteric inflammation in broiler chicks." Poultry Science 94, no. 6 (June 2015): 1353–59. http://dx.doi.org/10.3382/ps/pev111.
Повний текст джерела
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Maguey-Gonzalez, Jesus A., Matias A. Michel, Mikayla F. A. Baxter, Bruno Solis-Cruz, Daniel Hernandez-Patlan, Ruben Merino-Guzman, Xochitl Hernandez-Velasco, et al. "Effects of Humic Acids on Recovery of Salmonella Enterica Serovar Enteritidis." Annals of Animal Science 18, no. 2 (May 1, 2018): 387–99. http://dx.doi.org/10.1515/aoas-2017-0037.
Повний текст джерелаАнотація:
AbstractTwo experiments were conducted to evaluate the effects of humic acids (HA) on recovery of Salmonella Enteritidis, in an in vitro digestive system and on intestinal colonization in neonate broiler chickens. In experiment 1, two runs using an in vitro digestion model with two sources of HA (commercial or natural extraction) at 0.1 or 0.2%, and inoculated with 107 CFU/tube of S. Enteritidis, were carried out. In experiment 2, one-day-old male broiler chickens were randomly allocated to one of two groups (n=25) with or without 0.2% of isolated HA from worm compost, and challenged with 106 CFU of S. Enteritidis per chicken at 10-d old. All chicks were euthanized 24-h post challenge, and were subjected to serum fluorescein isothiocyanate dextran (FITC-d) determination. A section of ileum was removed to obtain total concentration of IgA. Ceca-cecal tonsils were removed to evaluate Salmonella recovery, total lactic acid bacteria (LAB) and total Gram negative bacteria. In experiment 1, neither concentration of commercial nor natural HA were able to reduce the recovery of S. Enteritidis in any of the simulated compartments (P>0.05). Only the crop compartment showed significant differences in pH in both trials between control and treated groups. In experiment 2, no significant differences were observed in serum concentration of FITC-d, intestinal IgA, S. Enteritidis recovery, LAB or total Gram negative bacteria in the ceca between control and treated chickens. In conclusion, no effects of HA on recovery of Salmonella Enteritidis, in an in vitro digestive system and on intestinal colonization of Salmonella, bacterial counts in ceca, intestinal IgA and serum FITC-d in neonate broiler chickens were observed. Further studies to evaluate the effect of HA under feed restriction model as an inducer of intestinal inflammation are currently being conducted.
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Silva, Francisco Laurindo da, Raphael Sanzio Pimenta, Juliana Fonseca Moreira da Silva, Déborah Aparecida Negrão Corrêa, and Ary Corrêa Junior. "The influence of carbohydrates in the interaction of Paracoccidioides brasiliensis with CCL-6 cells in vitro." Revista da Sociedade Brasileira de Medicina Tropical 45, no. 6 (December 2012): 739–44. http://dx.doi.org/10.1590/s0037-86822012000600016.
Повний текст джерелаАнотація:
INTRODUCTION: Little is known about the early events in the interaction between Paracoccidioides brasiliensis and its host. To understand the effect of carbohydrates in the interaction between the fungus and epithelial cell in culture, we analyzed the influence of different carbohydrate solutions on the adhesion of P. brasiliensis yeast cells to CCL-6 cells in culture. METHODS: Fungal cells were cultivated with the epithelial cell line, and different concentrations of D-fucose, N-acetyl-glucosamine, D-mannose, D-glucosamine, D-galactosamine, sorbitol and fructose were added at the beginning of the experiment. Six hours after the treatment, the cells were fixed and observed by light microscopy. The number of P. brasiliensis cells that were adhered to the CCL-6 monolayer was estimated. RESULTS: The number of adhesion events was diminished following treatments with D-fucose, N-acetyl-glucosamine, D-mannose, D-glucosamine and D-galactosamine as compared to the untreated controls. Sorbitol and fructose-treated cells had the same adhesion behavior as the observed in the control. P. brasiliensis propagules were treated with fluorescent lectins. The FITC-labeled lectins WGA and Con-A bound to P. brasiliensis yeast cells, while SBA and PNA did not. CONCLUSIONS: The perceptual of adhesion between P. brasiliensis and CCL-6 cells decreased with the use of D-mannose, N-acetyl-glucosamine and D-glucosamine. The assay using FITC-labeled lectins suggests the presence of N-acetyl-glucosamine, α-mannose and α-glucose on the P. brasiliensis cell surface. An enhanced knowledge of the mediators of adhesion on P. brasiliensis could be useful in the future for the development of more efficient and less harmful methods for disease treatment and control.
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Hao, Yuzhi, Xiaoyan Yang, Yongzhong Shi, Steven Song, James Xing, Janet Marowitch, Jianmin Chen, and Jie Chen. "Magnetic gold nanoparticles as a vehicle for fluorescein isothiocyanate and DNA delivery into plant cells." Botany 91, no. 7 (July 2013): 457–66. http://dx.doi.org/10.1139/cjb-2012-0281.
Повний текст джерелаАнотація:
Magnetic gold nanoparticles (mGNPs) with uniform size and morphology synthesized by our sonication treatment method were covalently bound with fluorescein isothiocyanate (FITC) molecules. Driven by an external magnetic field, FITC-labelled nanoparticles were delivered into plant cells with and without cell walls, evident from sectional transmission electron microscopy images. Confocal images further indicate that the green fluorescence in canola protoplasts and walled cells indeed came from the FITC molecules, instead of the chloroplasts’ autofluorescence. FITC-labelled nanoparticles had a delivery efficiency of 95% based on confocal images. In further study, plasmids were covalently bound with mGNPs, and delivered into canola cells with and without cell walls. After culturing for 48 h followed by staining with 5-bromo-4-chloro-3-indolyl-β-d-glucuronic acid (X-Gluc), blue colour appeared in the protoplasts, while the walled canola cells showed a green colour that can be interpreted as the combination of blue and yellow from the suspension cells themselves. The presence of the blue colour indicates the expression of the GUS gene; therefore, the plasmids were successfully delivered into the canola cells. Furthermore, on examination, mGNPs were considered to be noncytotoxic by fluorescein diacetate staining.
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Burns, A. R., J. Van Oostdam, D. C. Walker, and J. C. Hogg. "Effect of urethan anesthesia on cigarette smoke-induced airway injury in guinea pigs." Journal of Applied Physiology 63, no. 1 (July 1, 1987): 84–91. http://dx.doi.org/10.1152/jappl.1987.63.1.84.
Повний текст джерелаАнотація:
The effect of urethan anesthesia on cigarette smoke-induced airway responsiveness and permeability was studied in the guinea pig. Airway responsiveness was determined by measuring changes to airway resistance to graded doses of aerosolized histamine, and mucosal permeability was determined by measuring the appearance of fluorescein isothiocyanate-dextran (FITC-D) in the blood and examining its distribution in lung tissue after it had been delivered to the lung in an aerosol. The results confirm previous studies that smoke exposure increased airway responsiveness and mucosal permeability. They also show that urethan anesthesia administered before smoke exposure prevented the smoke-related changes in airway reactivity and mucosal permeability. In animals that remained conscious during the smoke exposure, there was increased deposition of the dextran in the regions of the bronchioloalveolar junctions with a more rapid uptake of FITC-D into the blood. We postulate that, when urethan anesthesia is administered before smoke exposure, the exudative phase of the inflammatory reaction produced by smoke exposure is suppressed.
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Sender, S., G. Gros, A. Waheed, G. S. Hageman, and W. S. Sly. "Immunohistochemical localization of carbonic anhydrase IV in capillaries of rat and human skeletal muscle." Journal of Histochemistry & Cytochemistry 42, no. 9 (September 1994): 1229–36. http://dx.doi.org/10.1177/42.9.8064130.
Повний текст джерелаАнотація:
We used polyclonal antisera raised in rabbits against membrane-bound rat lung and human lung carbonic anhydrase (CA) IV in immunofluorescence studies to stain cryosections of rat soleus and extensor digitorum longus (EDL) and several human skeletal muscles. There was strong specific staining of capillaries in all muscles investigated. Several techniques were applied to verify this result. (a) Serial sections were either incubated with anti-CA IV/FITC or processed for endothelial ATPase reaction. There was precise co-localization of antibody marked structures and ATPase stained capillaries. (b) Human muscle sections were double stained with anti-CA IV/TRITC and anti-von Willebrand factor (vWF)/FITC. vWF, a capillary marker, and CA IV were localized at identical sites. (c) The CAIV was released from capillaries by treatment with phosphatidylinositol specific phospholipase C, suggesting that the enzyme is anchored to the endothelial cell membrane via a phosphatidylinositolglycan anchor. (d) A rat hindlimb was perfused with diluted antiserum. Cryosections of perfused soleus and EDL processed for anti-rabbit IgG/FITC staining showed clear fluorescence associated with capillaries, indicating that the antigen was accessible from the capillary lumen. (e) Immune complexes formed during antiserum perfusion as described in d were precipitated from muscle homogenates. SDS-PAGE followed by immunoblotting showed that the predominant portion of total muscle CA IV was bound in these complexes and therefore must be located intravascularly.
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Coles, Makenly E., Aaron J. Forga, Roberto Señas-Cuesta, Brittany D. Graham, Callie M. Selby, Álvaro J. Uribe, Blanca C. Martínez, et al. "Assessment of Lippia origanoides Essential Oils in a Salmonella typhimurium, Eimeria maxima, and Clostridium perfringens Challenge Model to Induce Necrotic Enteritis in Broiler Chickens." Animals 11, no. 4 (April 13, 2021): 1111. http://dx.doi.org/10.3390/ani11041111.
Повний текст джерелаАнотація:
The objective of the present research was to evaluate dietary supplementation of essential oils from Lippia origanoides (LEO) on necrotic enteritis (NE). Chickens were randomly assigned to three groups. Group 1: negative control; Group 2: positive control challenged with Salmonella typhimurium (day 1), Eimeria maxima (day 18), and C. perfringens (CP, days 22-23); Group 3: dietary supplementation LEO and challenged. On d 25 of age, serum samples were collected to evaluate fluorescein isothiocyanate-dextran (FITC-d), superoxide dismutase (SOD), gamma interferon (IFN-γ), Immunoglobulin A (IgA). Group 3 showed a significant reduction of the harmful effects of induced infection/dysbiosis and a significant reduction in NE lesion scores, morbidity and mortality compared with the positive challenge control group (p < 0.05) compared with Group 2. Digested feed supernatant, supplemented with LEO and inoculated with CP, reduced CP burden (p < 0.05). Group 3 also exhibited a significant reduction in FITC-d, IFN-γ and IgA compared with Group 2. However, a significant increase SOD was observed in Group 3 compared with both control groups. Further investigation to compare the effect of LEO and the standard treatment of clostridial NE is required.
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Thornton, B. P., V. Vĕtvicka, M. Pitman, R. C. Goldman, and G. D. Ross. "Analysis of the sugar specificity and molecular location of the beta-glucan-binding lectin site of complement receptor type 3 (CD11b/CD18)." Journal of Immunology 156, no. 3 (February 1, 1996): 1235–46. http://dx.doi.org/10.4049/jimmunol.156.3.1235.
Повний текст джерелаАнотація:
Abstract Zymosan, the cell wall from Saccharomyces cerevisiae, was reported to be a macrophage activator through its beta-glucan over 30 yr ago. Nevertheless, the identity of the beta-glucan receptor has been controversial. This study showed that the alpha M beta 2-integrin, CR3 (Mac-1, CD11b/CD18) served as the beta-glucan receptor through one or more lectin sites located outside of the CD11b I-domain that contains the binding sites for iC3b, ICAM-1, and fibrinogen. Sugar specificity, analyzed with FITC-labeled soluble polysaccharides and flow cytometry, showed CR3-specific staining with several pure beta-glucans but not with alpha-mannan. However, a 10-kDa soluble zymosan polysaccharide (SZP) with high affinity (6.7 x 10(-8) M) for CR3 consisted largely of mannose and approximately 5% glucose. Binding of either SZP-FITC or beta-glucan-FITC to CR3 was blocked not only by pure beta-glucans from yeast, mushroom, seaweed, or barley, but also by N-acetyl-D-glucosamine (NADG), alpha- or beta-methylmannoside, and alpha- or beta-methyl-glucoside. SZP-FITC and beta-glucan-FITC stained all leukocyte types similarly to anti-CR3-FITC, and polysaccharide-FITC staining was inhibited > or = 95% by unlabeled anti-CR3. SZP-FITC staining of cells expressing recombinant chimeras between CR3 and CR4 (p150,95, CD11c/CD18) suggested that both the divalent cation-binding region of CD11b and the region C-terminal to it may regulate binding of polysaccharides to CR3. Unlabeled SZP or beta-glucan also blocked CR3 staining by 11 mAb to C-terminal domain epitopes of CD11b but had no effect on staining by mAb directed to the I-domain. In conclusion, CR3 serves as the leukocyte beta-glucan receptor through a cation-independent lectin site located C-terminal to the I-domain of CD11b. Its sugar specificity is broader than originally appreciated, allowing it to react with certain polysaccharides containing mannose or NADG, as well as glucose.
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FOUCAULT, Georges, Monique VACHER, Sophie CRIBIER та Martine ARRIO-DUPONT. "Interactions between β-enolase and creatine kinase in the cytosol of skeletal muscle cells". Biochemical Journal 346, № 1 (8 лютого 2000): 127–31. http://dx.doi.org/10.1042/bj3460127.
Повний текст джерелаАнотація:
We studied interactions in vivo between the cytosolic muscle isoform of creatine kinase (M-CK) and the muscle isoform of 2-phospho-D-glycerate hydrolyase (β-enolase) in muscle sarcoplasm by incubating glycerol-skinned fibres with FITC-labelled β-enolase in the presence or absence of free CK. A small amount of bound β-enolase was observed in the presence of large concentrations of CK. The mobility of enolase was measured in cultured satellite cells by modulated-fringe-pattern photobleaching. FITC-labelled β-enolase was totally mobile in both the presence and the absence of CK but its diffusion coefficient was slightly lower in the presence of CK. This suggests a weak interaction in vivo between enolase and CK.
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Kim, Evgenii, Jared Van Reet, Hyun-Chul Kim, Kavin Kowsari, and Seung-Schik Yoo. "High Incidence of Intracerebral Hemorrhaging Associated with the Application of Low-Intensity Focused Ultrasound Following Acute Cerebrovascular Injury by Intracortical Injection." Pharmaceutics 14, no. 10 (October 6, 2022): 2120. http://dx.doi.org/10.3390/pharmaceutics14102120.
Повний текст джерелаАнотація:
Low-intensity transcranial focused ultrasound (FUS) has gained momentum as a non-/minimally-invasive modality that facilitates the delivery of various pharmaceutical agents to the brain. With the additional ability to modulate regional brain tissue excitability, FUS is anticipated to confer potential neurotherapeutic applications whereby a deeper insight of its safety is warranted. We investigated the effects of FUS applied to the rat brain (Sprague-Dawley) shortly after an intracortical injection of fluorescent interstitial solutes, a widely used convection-enhanced delivery technique that directly (i.e., bypassing the blood–brain-barrier (BBB)) introduces drugs or interstitial tracers to the brain parenchyma. Texas Red ovalbumin (OA) and fluorescein isothiocyanate-dextran (FITC-d) were used as the interstitial tracers. Rats that did not receive sonication showed an expected interstitial distribution of OA and FITC-d around the injection site, with a wider volume distribution of OA (21.8 ± 4.0 µL) compared to that of FITC-d (7.8 ± 2.7 µL). Remarkably, nearly half of the rats exposed to the FUS developed intracerebral hemorrhaging (ICH), with a significantly higher volume of bleeding compared to a minor red blood cell extravasation from the animals that were not exposed to sonication. This finding suggests that the local cerebrovascular injury inflicted by the micro-injection was further exacerbated by the application of sonication, particularly during the acute stage of injury. Smaller tracer volume distributions and weaker fluorescent intensities, compared to the unsonicated animals, were observed for the sonicated rats that did not manifest hemorrhaging, which may indicate an enhanced degree of clearance of the injected tracers. Our results call for careful safety precautions when ultrasound sonication is desired among groups under elevated risks associated with a weakened or damaged vascular integrity.
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Lubenko, Anatole, John Raymond Collier, Mark Williams, Damien Hindmarch, Sally Rosemary Wilson, and Julie Pluck. "Quantitating fetomaternal hemorrhages of D+ red cells using an FITC-conjugated IgG monoclonal anti-D by flow cytometry: a case report." Immunohematology 13, no. 1 (2020): 12–14. http://dx.doi.org/10.21307/immunohematology-2019-693.
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Komane, Patrick P., Pradeep Kumar, and Yahya E. Choonara. "Atrial Natriuretic Peptide Antibody-Functionalised, PEGylated Multiwalled Carbon Nanotubes for Targeted Ischemic Stroke Intervention." Pharmaceutics 13, no. 9 (August 28, 2021): 1357. http://dx.doi.org/10.3390/pharmaceutics13091357.
Повний текст джерелаАнотація:
Stroke is one of the major causes of disability and the second major cause of death around the globe. There is a dire need for an ultrasensitive detection tool and an effective and efficient therapeutic system for both detection and treatment of stroke at its infancy stage. Carbon nanotubes are promising nanomaterials for tackling these challenges. The loading of dexamethasone and decoration of PEGylated multiwalled carbon nanotube with atrial natriuretic peptide (ANP) antibody and fluorescein isothiocyanate for targeting ischemic site in the rat stroke model is presented here. Functionalisation of carbon nanotubes with dexamethasone (DEX), polyethylene glycol (PEG), fluorescein isothiocyanate (FITC), and ANP antibody caused a 63-fold increase in the D band intensity as illustrated by Raman. The characteristic band intensity increase was observed at 1636 nm following functionalisation of carbon nanotubes with polyethylene glycol and dexamethasone as confirmed by Fourier Transform Infrared. These findings have demonstrated the coupling capability of atrial natriuretic peptide antibody to DEX-PEG-CNTs. The baseline plasma atrial natriuretic peptide levels were ranging from 118 to 135.70 pg/mL prior to surgery and from 522.09 to 552.37 following common carotid artery occlusion. A decrease in atrial natriuretic peptide levels to 307.77 was observed when the rats were treated with FITC-DEX-PEG-ANP-CNTs, PEG-CNTs and DEX with a significant drop in the FITC-DEX-PEG-ANP-CNTs treated group. Fluorescence was detected in FITC-DEX-PEG-CNTs and FITC-DEX-PEG-ANP-CNTs treated ischemic stroke rats. The highest fluorescence intensity was reported in plasma (2179) followed by the kidney (1563) and liver (1507). These findings suggest a beneficial role that is played by the FITC-DEX-PEG-ANP-CNTs in the reduction of inflammation in the ischemic stroke induced rats that could induce a successful treatment of ischemic stroke.
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TSUJIE, MICHIKO, YOSHITAKA ISAKA, HIROYUKI NAKAMURA, ENYU IMAI, and MASATSUGU HORI. "Electroporation-Mediated Gene Transfer that Targets Glomeruli." Journal of the American Society of Nephrology 12, no. 5 (May 2001): 949–54. http://dx.doi.org/10.1681/asn.v125949.
Повний текст джерелаАнотація:
Abstract. Electroporation has been applied to introducing DNA into several organs; however, gene expression was localized around the injected area. Examined was the efficiency of intrarenal injection of DNA followed by in vivo electroporation, using FITC-labeled oligodeoxynucleotides (FITC-ODN) and plasmid DNA expressing β-galactosidase or luciferase. FITC-ODN or expression vectors were injected into the left renal artery; thereafter, the left kidney was electroporated between a pair of tweezer-type electrodes. FITC-ODN were transferred into all glomeruli, and transfected cells were identified as mesangial cells. Four d after transfection of the pCAGGS-LacZ gene, β-galactosidase expression was observed in 75% of glomeruli. To compare the transfection efficacy by electroporation with that by the hemagglutinating virus of Japan (HVJ) liposome method, a luciferase reporter gene, pActLuc, was transferred into glomeruli by either electroporation or the HVJ liposome method. On day 4, electroporation resulted in higher glomerular luciferase activity than did the HVJ liposome method. We also observed that co-transfection of pcEBNA, an expression vector for Epstein-Barr virus nuclear antigen, and poriP-cLuc, oriP-harboring vector, resulted in an eightfold higher luciferase gene expression than simple poriP-cLuc. No histologic damages were seen in glomeruli or tubular epithelial cells. In conclusion, gene transfer into renal artery followed by electroporation was an effective and simple strategy for gene transfer that targets glomerular mesangial cells.
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Zubair, Muhammad Sulaiman, Walied Mohamed Alarif, Mohamed Ali Ghandourah, Syariful Anam та Ibrahim Jantan. "Cytotoxic Activity of 2-O-β-glucopyranosil Cucurbitacin D from Benalu Batu (Begonia sp.) Growing in Morowali, Central Sulawesi". Indonesian Journal of Chemistry 20, № 4 (10 червня 2020): 766. http://dx.doi.org/10.22146/ijc.43626.
Повний текст джерелаАнотація:
Benalu batu (Begonia sp.) had been used traditionally as an anticancer medicinal plant by Wana tribe in Morowali, Central Sulawesi, This study aims to evaluate the cytotoxic activity of 2-O-β-glucopyranosil cucurbitacin D, isolated from the ethyl acetate soluble fraction of Benalu batu (Begonia sp.) and to determine its action on apoptosis induction. Benalu batu (Begonia sp.) herb was extracted by maceration using ethanol 96% as a solvent. Vacuum liquid column chromatography and preparative thin layer chromatography have been applied on fractionation and isolation of the compound. The structure elucidation was performed by extensive analysis of 1D/2D nuclear magnetic resonance (NMR) and Mass Spectrophotometer (MS). Cytotoxic activity against human breast adenocarcinoma (MCF-7) and human colon colorectal carcinoma (HCT-116) cell lines were performed by 5-diphenyltetrazolium bromide (MTT) method. Annexin V-FITC assay was employed to determine the apoptosis induction. 2-O-β-glucopyranosil cucurbitacin D showed potent cytotoxic activity against MCF-7 and HCT-116 with the IC50 of 19.913 and 0.002 μg/mL, respectively. Annexin V-FITC assay clearly exhibited the cytotoxic mechanism on MCF-7 and HCT-116 via apoptosis induction with a significant percentage of early and late apoptosis of 75.8 and 78.4%, respectively. This study reveals the potential cytotoxic activity of 2-O-β-glucopyranosil cucurbitacin D isolated from Benalu batu and its mechanism via apoptosis induction.
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Liu, Jundi, Po-Yun Teng, Woo K. Kim, and Todd J. Applegate. "Assay considerations for fluorescein isothiocyanate-dextran (FITC-d): an indicator of intestinal permeability in broiler chickens." Poultry Science 100, no. 7 (July 2021): 101202. http://dx.doi.org/10.1016/j.psj.2021.101202.
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Su, Lin, Rui Wu, Xinrou Chen, Wei Hou, and Benfang Helen Ruan. "FITC-labeled d-glucose analog is suitable as a probe for detecting insulin-dependent glucose uptake." Bioorganic & Medicinal Chemistry Letters 28, no. 22 (December 2018): 3560–63. http://dx.doi.org/10.1016/j.bmcl.2018.09.027.
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Al-Naemi, Hamda, and Ann L. Baldwin. "Nitric oxide: role in venular permeability recovery after histamine challenge." American Journal of Physiology-Heart and Circulatory Physiology 277, no. 5 (November 1, 1999): H2010—H2016. http://dx.doi.org/10.1152/ajpheart.1999.277.5.h2010.
Повний текст джерелаАнотація:
Histamine is an inflammatory mediator produced by mast cells that reside close to blood vessels. It causes a transient increase in venular permeability and stimulates endothelial production of nitric oxide (NO). In this study, we investigated the role that NO plays in the permeability recovery and evaluated the response of mast cells. The mesenteric microvasculature of anesthetized rats was suffused with 10−3 M histamine for 3 min and then perfused with the NO donor sodium nitroprusside (SNP; 10−6 M), the NO inhibitor N G-monomethyl-l-arginine (l-NMMA; 10−5 M), its enantiomer (d-NMMA; 10−5 M), or HEPES-buffered saline containing 0.5% BSA for 15 min. This was replaced by FITC-albumin for 3 min, followed by fixative. The vasculature was visualized using epifluorescence microscopy and was stained for mast cells. Preparations treated with histamine only showed discrete FITC-albumin leaks. Subsequent inhibition of NO increased venular FITC-albumin leaks and prevented permeability recovery, whereas subsequent treatment with SNP decreased the histamine-induced venular leaks. Mast cells degranulated due to histamine and the other treatment combinations. In conclusion, inhibition of NO prevented permeability recovery and depleted mast cells of their histamine content.
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Choi, Young-A., R. Tao, K. Yonemori, and A. Sugiura. "Multi-color Genomic In Situ Hybridization Identifies Parental Chromosomes in Somatic Hybrids of Diospyros kaki and D. glandulosa." HortScience 37, no. 1 (February 2002): 184–86. http://dx.doi.org/10.21273/hortsci.37.1.184.
Повний текст джерелаАнотація:
Multi-color genomic in situ hybridization (MCGISH) was performed for mitotic cells of the somatic hybrids of Diospyros kaki (2n = 6x = 90) and D. glandulosa (2n = 2x = 30). Total DNA of D. kaki and D. glandulosa were isolated and labeled with biotin-16-UTP and digoxigenin (DIG)-11-UTP, respectively. The labeled DNAs were used as probes to differentiate parental chromosomes. The biotin-labeled probe was detected with avidin-rhodamine, and the DIG-labeled probe was detected with anti-DIG-FITC (fluorescein isothiocyanate). Ninety chromosomes from D. kaki that showed reddish-orange and 30 chromosomes from D. glandulosa that showed greenish-yellow were observed under a fluorescence microscope. Some chromosomes showed cross-hybridization with both probes at their terminal or other chromosome regions. These results indicated that MCGISH could be used to analyze genomes of Diospyros species whose chromosomes are small and numerous.
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Chen, Shan-Na, Ying-Xue Ma, Song Chen, Guang-Hui He, Mei Han, Xiang Gao, Jun-Hua Wang, Bin Wu, and Jian Wang. "Protective effect of LIF-huMSCs on the retina of diabetic model rats." International Journal of Ophthalmology 14, no. 10 (October 18, 2021): 1508–17. http://dx.doi.org/10.18240/ijo.2021.10.06.
Повний текст джерелаАнотація:
AIM: To investigate the protective effect of human umbilical cord mesenchymal stem cells (hUCMSCs) modified by the LIF gene on the retinal function of diabetic model rats and preliminarily explore the possible mechanism. METHODS: A stably transfected cell line of hUCMSCs overexpressing leukemia inhibitory factor (LIF) was constructed. Overexpression was verified by fluorescent quantitative polymerase chain reaction (qPCR). Forty-eight adult Sprague-Dawley rats were randomly divided into a normal control group (group A), streptozotocin-induced diabetic control group (group B), diabetic rats at 3mo injected with empty vector-transfected hUCMSCs (group C) or injected with LIF-hUCMSCs (group D). Four weeks after the intravitreal injection, analyses in all groups included retinal function using flash electroretinogram (F-ERG), retinal blood vessel examination of retinal flat mounts perfused with fluorescein isothiocyanate-dextran (FITC-dextran), and retinal structure examination of sections using hematoxylin and eosin staining. Expression levels of adiponectin (APN), high-sensitivity C-reactive protein (hs-CRP), and neurotrophin-4 (NT-4) in each group was detected using immunohistochemistry, PCR, Western blotting, and ELISA, respectively. RESULTS: A stable transgenic cell line of LIF-hUCMSCs was constructed. F-ERG and FITC-dextran examinations revealed no abnormalities of retinal structure and function in group A, severe damage of the retinal blood vessels and function in group B, and improved retinal structure and function in group C and especially group D. qPCR, ELISA, and Western blot analyses revealed progressively higher APN and NT-4 expression levels in groups B, C, and D than in group A. hs-CRP expression was significantly higher in group B than in groups A, C, and D, and was significantly higher in group C than in group D (P<0.05). CONCLUSION: LIF-hUCMSCs protect the retina of diabetic rats by upregulating APN and NT-4 expression and downregulating hs-CRP expression in the retina.
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Gao, Chunli, Simin Tang, Haixing Zhang, Huishu Zhang, Tian Zhang, Bin Bao, Yuping Zhu, and Wenhui Wu. "A Novel Marine Pyran-Isoindolone Compound Enhances Fibrin Lysis Mediated by Single-Chain Urokinase-Type Plasminogen Activator." Marine Drugs 20, no. 8 (July 30, 2022): 495. http://dx.doi.org/10.3390/md20080495.
Повний текст джерелаАнотація:
Fungi fibrinolytic compound 1 (FGFC1) is a rare pyran-isoindolone derivative with fibrinolytic activity. The aim of this study was to further determine the effect of FGFC1 on fibrin clots lysis in vitro. We constructed a fibrinolytic system containing single-chain urokinase-type plasminogen activator (scu-PA) and plasminogen to measure the fibrinolytic activity of FGFC1 using the chromogenic substrate method. After FITC-fibrin was incubated with increasing concentrations of FGFC1, the changes in the fluorescence intensity and D-dimer in the lysate were measured using a fluorescence microplate reader. The fibrin clot structure induced by FGFC1 was observed and analyzed using a scanning electron microscope and laser confocal microscope. We found that the chromogenic reaction rate of the mixture system increased from (15.9 ± 1.51) × 10−3 min−1 in the control group to (29.7 ± 1.25) × 10−3 min−1 for 12.8 μM FGFC1(p < 0.01). FGFC1 also significantly increased the fluorescence intensity and d-dimer concentration in FITC fibrin lysate. Image analysis showed that FGFC1 significantly reduced the fiber density and increased the fiber diameter and the distance between protofibrils. These results show that FGFC1 can effectively promote fibrin lysis in vitro and may represent a novel candidate agent for thrombolytic therapy.
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Duff, Audrey F., Mikayla F. A. Baxter, B. Danielle Graham, Billy M. Hargis, and Lisa R. Bielke. "Mode of Action of Dietary Dexamethasone May Not Be Dependent Upon Microbial Mechanisms in Broilers." Microorganisms 7, no. 9 (September 12, 2019): 346. http://dx.doi.org/10.3390/microorganisms7090346.
Повний текст джерелаАнотація:
Dexamethasone (Dex), a synthetic glucocorticoid (GC), in feed has been shown to increase gut permeability via stress-mediated mechanisms, but the exact mode of action on gut barrier function is not fully understood. Stress has been reported to alter the profile and virulence of intestinal flora predisposing for opportunistic disease. This study aimed to evaluate the relationship between dietary Dex and recoverable intestinal microbial profile in broilers to better understand mode of action and refine future uses of the model. Three experiments were conducted that administered Dex-treated feed for one week in conjunction with the antibiotics BMD (bacitracin methylene disalicylate) or Baytril® (enrofloxacin) to evaluate if enteric microbial mechanisms were important in Dex-induced permeability. Serum fluorescein isothiocyanate-dextran (FITC-d) and bacterial translocation (BT) have been reported to increase after Dex treatment and were used to assess gut epithelial leakage. Shifts in bacterial profiles were also measured on selective agar. Combining Dex with BMD or Baytril resulted in increased (P < 0.05) serum FITC-d versus Dex-only. Additionally, Baytril did not reduce aerobic BT and bacterial profiles remained similar after Dex. These results suggest a minimal role of intestinal microbes in Dex-induced changes to intestinal barrier function.
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Hernandez-Patlan, Daniel, Bruno Solis-Cruz, Karine P. Pontin, Juan D. Latorre, Xochitl Hernandez-Velasco, Ruben Merino-Guzman, Abraham Mendez-Albores, Billy M. Hargis, Raquel Lopez-Arellano, and Guillermo Tellez-Isaias. "Evaluation of Ascorbic Acid or Curcumin Formulated in a Solid Dispersion on Salmonella Enteritidis Infection and Intestinal Integrity in Broiler Chickens." Pathogens 8, no. 4 (November 10, 2019): 229. http://dx.doi.org/10.3390/pathogens8040229.
Повний текст джерелаАнотація:
Two experimental models were conducted to evaluate and compare the effect of ascorbic acid (AA) or curcumin formulated in a solid dispersion (SD-CUR) as prophylactic or therapeutic alternatives to prevent or control S. Enteritidis (SE) infection in broiler chickens. In the prophylactic model, dietary administration of AA showed a significant reduction in SE counts in crop compared to the positive control (PC) group (p < 0.05), whereas in cecal tonsils (CT), SD-CUR significantly reduced SE recovery. Superoxide dismutase (SOD) activity was significantly higher in chickens supplemented with AA or SD-CUR, and total intestinal IgA levels were significantly lower in both treatments when compared to the PC group. Serum fluorescein isothiocyanate-dextran (FITC-d) levels were reduced by SD-CUR compared to PC, while AA presented significantly lower total aerobic bacteria. In the therapeutic model, only the dietary administration of AA significantly decreased SE in crop and CT on days 3 and 10 post-challenge. FITC-d levels were significantly lower in both treated groups in comparison to PC, but IgA levels were significantly reduced only by AA. The results suggest that dietary AA and SD-CUR have different modes of action to reduce SE intestinal colonization in two different challenge models in broiler chickens.
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Axelsson, Josefin, Kristinn Sverrisson, Anna Rippe, William Fissell, and Bengt Rippe. "Reduced diffusion of charge-modified, conformationally intact anionic Ficoll relative to neutral Ficoll across the rat glomerular filtration barrier in vivo." American Journal of Physiology-Renal Physiology 301, no. 4 (October 2011): F708—F712. http://dx.doi.org/10.1152/ajprenal.00183.2011.
Повний текст джерелаАнотація:
The glomerular filtration barrier (GFB) is commonly conceived as a negatively charged sieve to proteins. Recent studies, however, indicate that glomerular charge effects are small for anionic, carboxymethylated (CM) dextran vs. neutral dextran. Furthermore, two studies assessing the glomerular sieving coefficients (θ) for negative CM-Ficoll vs. native Ficoll have demonstrated an increased glomerular permeability for CM-Ficoll (Asgeirsson D, Venturoli D, Rippe B, Rippe C. Am J Physiol Renal Physiol 291: F1083–F1089, 2006; Guimarães M, Nikolovski J, Pratt L, Greive K, Comper W. Am Physiol Renal Physiol 285: F1118–F1124, 2003.). The CM-Ficoll used, however, showed a larger Stokes-Einstein radius ( ae) than neutral Ficoll, and it was proposed that the introduction of negative charges in the Ficoll molecule had made it more flexible and permeable. Recently, a negative FITC-labeled CM-Ficoll (CMI-Ficoll) was produced with a conformation identical to that of neutral FITC-Ficoll. Using these probes, we determined their θ:s in anesthetized Wistar rats (259 ± 2.5 g). After blood access had been achieved, the left ureter was cannulated for urine sampling. Either polysaccharide was infused (iv) together with a filtration marker, and urine and plasma were collected. Assessment of θ FITC-Ficoll was achieved by high-performance size-exclusion chromatography (HPSEC). CMI-Ficoll and native Ficoll had identical elugrams on the HPSEC. Diffusion of anionic Ficoll was significantly reduced compared with that of neutral Ficoll across the GFB for molecules of ae ∼20–35 Å, while there were no charge effects for Ficoll of ae = 35–80 Å. The data are consistent with a charge effect present in “small pores,” but not in “large pores,” of the GFB and mimicked those obtained for anionic membranes in vitro for the same probes.
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Beasley, Selina, Mohamed El-Sherbiny, Sylvia Megyerdi, Sally El-Shafey, Karishma Choksi, Ismail Kaddour-Djebbar, Nader Sheibani, Stephen Hsu, and Mohamed Al-Shabrawey. "Caspase-14 Expression Impairs Retinal Pigment Epithelium Barrier Function: Potential Role in Diabetic Macular Edema." BioMed Research International 2014 (2014): 1–11. http://dx.doi.org/10.1155/2014/417986.
Повний текст джерелаАнотація:
We recently showed that caspase-14 is a novel molecule in retina with potential role in accelerated vascular cell death during diabetic retinopathy (DR). Here, we evaluated whether caspase-14 is implicated in retinal pigment epithelial cells (RPE) dysfunction under hyperglycemia. The impact of high glucose (HG, 30 mM D-glucose) on caspase-14 expression in human RPE (ARPE-19) cells was tested, which showed significant increase in caspase-14 expression compared with normal glucose (5 mM D-glucose + 25 mM L-glucose). We also evaluated the impact of modulating caspase-14 expression on RPE cells barrier function, phagocytosis, and activation of other caspases using ARPE-19 cells transfected with caspase-14 plasmid or caspase-14 siRNA. We used FITC-dextran flux assay and electric cell substrate impedance sensing (ECIS) to test the changes in RPE cell barrier function. Similar to HG, caspase-14 expression in ARPE-19 cells increased FITC-dextran leakage through the confluent monolayer and decreased the transcellular electrical resistance (TER). These effects of HG were prevented by caspase-14 knockdown. Furthermore, caspase-14 knockdown prevented the HG-induced activation of caspase-1 and caspase-9, the only activated caspases by HG. Phagocytic activity was unaffected by caspase-14 expression. Our results suggest that caspase-14 contributes to RPE cell barrier disruption under hyperglycemic conditions and thus plays a role in the development of diabetic macular edema.
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Baldwin, Ann L., Gavin Thurston, and Hamda Al Naemi. "Inhibition of nitric oxide synthesis increases venular permeability and alters endothelial actin cytoskeleton." American Journal of Physiology-Heart and Circulatory Physiology 274, no. 5 (May 1, 1998): H1776—H1784. http://dx.doi.org/10.1152/ajpheart.1998.274.5.h1776.
Повний текст джерелаАнотація:
Inhibition of nitric oxide (NO) synthesis using N G-nitro-l-arginine methyl ester (l-NAME) or N G-monomethyl-l-arginine (l-NMMA) increases venular permeability in the rat mesentery (I. Kurose, R. Wolf, M. B. Grisham, T. Y. Aw, R. D. Specian, and D. N. Granger. Circ. Res. 76: 30–39, 1995), but the cellular mechanisms of this response are not known. This study was performed to determine whether such venular leaks are associated with changes in the endothelial actin cytoskeleton. In anesthetized Sprague-Dawley rats, the microvasculature of a mesenteric window was perfused with buffered saline, with or without 10−5M l-NAME,l-NMMA, or the inactive enantiomer N G-nitro-d-arginine methyl ester for 3 or 30 min. FITC-albumin was added to the perfusate for the last 3 min. The vasculature was perfusion fixed, stained for filamentous actin and for mast cells, and viewed microscopically. In control preparations, venules showed few FITC-albumin leaks and the endothelial actin cytoskeleton consisted of a peripheral rim along the cell-cell junctions. Preparations treated withl-NAME orl-NMMA showed significantly more leakage, the actin rims in leaky venules were discontinuous, and short, randomly oriented fibers appeared within the cells. In nonleaky venules, the peripheral actin rims sometimes contained small, equally spaced discontinuities not seen in control preparations. Although a mast cell stabilizer was used, 27–70% of the mast cells were degranulated in the presence ofl-NMMA. Thus inhibition of NO synthesis alters the endothelial cytoskeleton and increases albumin leakage from mesenteric venules, either directly or indirectly via the involvement of mast cells.
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Nakamura, Koji, Kozo Takayama, Tsuneji Nagai та Yoshie Maitani. "Regional Intestinal Absorption of FITC–Dextran 4,400 with Nanoparticles Based on β-Sitosterol β-D-Glucoside in Rats". Journal of Pharmaceutical Sciences 92, № 2 (лютий 2003): 311–18. http://dx.doi.org/10.1002/jps.10292.
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Cherukuri, Anu, Janie Frye, Todd French, Gary Durack, and Edward W. Voss. "FITC-poly-D-lysine conjugates as fluorescent probes to quantify hapten-specific macrophage receptor binding and uptake kinetics." Cytometry 31, no. 2 (February 1, 1998): 110–24. http://dx.doi.org/10.1002/(sici)1097-0320(19980201)31:2<110::aid-cyto6>3.0.co;2-q.
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Gupta, Reshu, Liangjie Yin, Astrid Grosche, Shanshan Lin, Xiaodong Xu, Jing Guo, Lauren A. Vaught, Paul G. Okunieff, and Sadasivan Vidyasagar. "An Amino Acid–Based Oral Rehydration Solution Regulates Radiation-Induced Intestinal Barrier Disruption in Mice." Journal of Nutrition 150, no. 5 (March 4, 2020): 1100–1108. http://dx.doi.org/10.1093/jn/nxaa025.
Повний текст джерелаАнотація:
ABSTRACT Background Radiotherapy inadvertently affects gastrointestinal (GI) epithelial cells, causing intestinal barrier disruption and increased permeability. Objective We examined the effect of amino acid–based oral rehydration solution (AA-ORS) on radiation-induced changes of intestinal barrier function and epithelial tight junctions (TJs) in a randomized experimental study using a total-body irradiation (TBI) mouse model. Methods Eight-week-old male Swiss mice received a single-dose TBI (0, 1, 3, or 5 Gy), and subsequent gastric gavage with AA-ORS (threonine, valine, serine, tyrosine, and aspartic acid) or saline for 2 or 6 d. Intestinal barrier function of mouse ileum was characterized by electrophysiological analysis of conductance, anion selectivity, and paracellular permeability [fluorescein isothiocyanate (FITC)-dextran]. Ultrastructural changes of TJs were evaluated by transmission electron microscopy. Membrane protein and mRNA expression of claudin-1, -2, -3, -5, and -7, occludin, and E-cadherin were analyzed with western blot, qPCR, and immunohistochemistry. Nonparametric tests were used to compare treatment-dose differences for each time point. Results Saline-treated mice had a higher conductance at doses as low as 3 Gy, and as early as 2 d post-TBI compared with 0 Gy (P < 0.001). Paracellular permeability and dilution potential were increased 6 d after 5 Gy TBI (P < 0.001). Conductance decreased with AA-ORS after 2 d in 3-Gy and 5-Gy mice (P < 0.05 and P < 0.001), and on day 6 after 5 Gy TBI (P < 0.001). Anion selectivity and FITC permeability decreased from 0.73 ± 0.02 to 0.61 ± 0.03 pCl/pNa (P < 0.01) and from 2.7 ± 0.1 × 105 to 2.1 ± 0.1 × 105 RFU (P < 0.001) in 5-Gy mice treated with AA-ORS for 6 d compared with saline. Irradiation-induced ultrastructural changes of TJs characterized by decreased electron density and gap formation improved with AA-ORS. Reduced claudin-1, -3, and -7 membrane expression after TBI recovered with AA-ORS within 6 d, whereas claudin-2 decreased indicating restitution of TJ proteins. Conclusions Radiation-induced functional and structural disruption of the intestinal barrier in mice is reversed by AA-ORS rendering AA-ORS a potential treatment option in prospective clinical trials in patients with gastrointestinal barrier dysfunction.
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Vink, A. A., F. M. Strickland, C. Bucana, P. A. Cox, L. Roza, D. B. Yarosh, and M. L. Kripke. "Localization of DNA damage and its role in altered antigen-presenting cell function in ultraviolet-irradiated mice." Journal of Experimental Medicine 183, no. 4 (April 1, 1996): 1491–500. http://dx.doi.org/10.1084/jem.183.4.1491.
Повний текст джерелаАнотація:
Prior ultraviolet (UV) irradiation of the site of application of hapten on murine skin reduces contact sensitization, impairs the ability of dendritic cells in the draining lymph nodes (DLN) to present antigen, and leads to development of hapten-specific suppressor T lymphocytes. We tested the hypothesis that UV-induced DNA damage plays a role in the impaired antigen-presenting activity of DLN cells. First, we assessed the location and persistence of cells containing DNA damage. A monoclonal antibody specific for cyclobutyl pyrimidine dimers (CPD) was used to identify UV-damaged cells in the skin and DLN of C3H mice exposed to UV radiation. Cells containing CPD were present in the epidermis, dermis, and DLN and persisted, particularly in the dermis, for at least 4 d after UV irradiation. When fluorescein isothiocyanate (FITC) was applied to UV-exposed skin, the DLN contained cells that were Ia+, FITC+, and CPD+; such cells from mice sensitized 3 d after UV irradiation exhibited reduced antigen-presenting function in vivo. We then assessed the role of DNA damage in UV-induced modulation of antigen-presenting cell (APC) function by using a novel method of increasing DNA repair in mouse skin in vivo. Liposomes containing T4 endonuclease V (T4N5) were applied to the site of UV exposure immediately after irradiation. This treatment prevented the impairment in APC function and reduced the number of CPD+ cells in the DLN of UV-irradiated mice. Treatment of unirradiated skin with T4N5 in liposomes or treatment of UV-irradiated skin with liposomes containing heat-inactivated T4N5 did not restore immune function. These studies demonstrate that cutaneous immune cells sustain DNA damage in vivo that persists for several days, and that FITC sensitization causes the migration of these to the DLN, which exhibits impaired APC function. Further, they support the hypothesis that DNA damage is an essential initiator of one or more of the steps involved in impaired APC function after UV irradiation.
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Ruff, Jared, Thaina L. Barros, Joy Campbell, Ricardo González-Esquerra, Christine N. Vuong, Sami Dridi, Elizabeth S. Greene, Xochitl Hernandez-Velasco, Billy M. Hargis, and Guillermo Tellez-Isaias. "Spray-Dried Plasma Improves Body Weight, Intestinal Barrier Function, and Tibia Strength during Experimental Constant Heat Stress Conditions." Animals 11, no. 8 (July 26, 2021): 2213. http://dx.doi.org/10.3390/ani11082213.
Повний текст джерелаАнотація:
The aim of this study was to see how spray-dried plasma (SDP) supplementation affected broiler chicken performance, intestinal permeability, and bone strength during persistent heat stress. One-day-old chicks (n = 480) were randomly assigned into twelve environmental corrals; four thermoneutral (TN-negative control, maintained at 24 °C from d 21–42); four heat stress (HS, exposed to 35 °C from d 21–42); and four heat stress treated with 2% SDP in the feed until d 28 followed by 1% SDP until d 42 (HS-SDP). The performance and serum levels of fluorescein isothiocyanate-dextran (FITC-d) were evaluated at d 21, 28, 35, and 42. The tibias strength was evaluated on d 21 and 42. The increment in chicken temperature (p < 0.05) was observed two h following the increase in environmental temperature in both HS groups and was associated with decreased performance parameters compared with the TN group. At d 42 of age, the chickens exposed to HS had an impaired gut permeability and decreased tibia strength compared to the TN group (p < 0.05). However, partially feeding SDP mitigated these adverse effects significantly. These findings imply that using SDP strategically during stressful times, such as prolonged heat stress, may help mitigate its negative consequences.
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Koller-Peroutka, Marianne, Stefanie Krammer, Anselm Pavlik, Manfred Edlinger, Ingeborg Lang, and Wolfram Adlassnig. "Endocytosis and Digestion in Carnivorous Pitcher Plants of the Family Sarraceniaceae." Plants 8, no. 10 (September 24, 2019): 367. http://dx.doi.org/10.3390/plants8100367.
Повний текст джерелаАнотація:
Highly evolved carnivorous plants secrete digestive enzymes for degradation of trapped animals and absorb whole macromolecules from their prey by means of endocytosis. (1) Background: In the pitcher-plant family Sarraceniaceae, the production of enzymes is dubious and no evidence for endocytosis is known so far. (2) Methods: Heliamphora nutans, Darlingtonia californica, and nine taxa of Sarracenia are tested for cuticular pores, and for protease and endocytosis of the fluorescent protein analogue FITC-BSA, after 10–48 h of stimulation. (3) Results: Cuticular pores as a prerequisite for enzyme secretion and nutrient uptake are present in all tested species. Permeable cells form clusters in the inner epidermis of the pitchers, but are only little differentiated from impermeable epidermis cells. Proteases are found in S. psittacina, S. leucophylla, S. minor, S. oreophila, S. alabamensis, H. nutans, D. californica lacking only in S. flava and in S. purpurea ssp. purpurea, S. purpurea ssp. venosa, S. rosea, where enzyme production is possibly replaced by degradation via the extraordinary diverse inquiline fauna. S. leucophylla, S. minor, S. oreophila exhibit both protease production and endocytosis; S. psittacina, S. alabamensis, H. nutans, D. californica produce proteases only; no single species shows endocytosis without protease production. (4) Conclusions: Protease secretion seems to be a prerequisite for endocytotic nutrient uptake. Transport of FITC-BSA absorbed by endocytosis towards the vascular tissue of the trap leaves suggests that endocytosis of nutrients is more than a side effect of enzyme secretion.
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Schmidt, Stefanie, Toralf Roch, Simi Mathew, Nan Ma, Christian Wischke, and Andreas Lendlein. "Correlating the Uptake and Dendritic Cell Activation by MDP-loaded Microparticles." MRS Proceedings 1569 (2013): 179–84. http://dx.doi.org/10.1557/opl.2013.816.
Повний текст джерелаАнотація:
ABSTRACTPolymer-based, degradable microparticles (MP) are attractive delivery vehicles for vaccines as the polymer properties can be specifically tailored and the carrier can be loaded with adjuvant. For all newly developed carrier systems it is important to analyze cellular uptake efficiency and the specific effects mediated by the encapsulated agent when phagocytosed by the cells, which is barely reported so far. By the encapsulation of N-acetylmuramyl-L-alanyl-D-isoglutamine (MDP) labeled with fluoresceinisothiocyanat (FITC) in poly[(rac-lactide)-co-glycolide] (PLGA) MP, the MP was fluorescent and used to visualize the phagocytic uptake. Since encapsulated MDP can activate dendritic cells (DC) via the cytosolic nucleotide-binding oligomerization domain receptors (NOD), it can be investigated whether only cells that have phagocytosed the MP are activated or whether bystander effects occur, resulting in activation of cells, which did not take up MDP-FITC loaded MP. Here, it is demonstrated that increasing MP concentrations in the culture medium had no impact on the viability of DC and that the MP uptake efficiency was dose dependent. Interestingly, it could be shown by the CD86 expression, that only DC, which had engulfed MP, were significantly stronger activated than DC, which had not phagocytosed MDP-FITC loaded MP. On the one hand these results indicate that sufficient amounts of MDP were released from the PLGA carriers into the cytosol of the DC. On the other hand, based on the correlation of uptake and activation on the single cell level, minimal MP induced bystander effects may be expected for in vivo applications.
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Blajman, J. E., D. M. Astesana, J. A. Zimmermann, E. Rossler, A. Romero Scharpen, A. P. Berisvil, M. V. Zbrun, L. P. Soto, M. R. Rosmini, and L. S. Frizzo. "Quantification of FITC-labelled probiotic Lactobacillus salivarius DSPV 001P during gastrointestinal transit in broilers." Beneficial Microbes 8, no. 1 (February 7, 2017): 55–64. http://dx.doi.org/10.3920/bm2016.0025.
Повний текст джерелаАнотація:
The knowledge related to the fate of probiotics in the complex environment of the intestinal microbiota in broilers is just beginning to be elucidated; however, it is not yet well understood. A good method to investigate the mechanisms by which probiotics mediate their effects is to mark probiotic bacteria and trace them. The aim of this research was to develop a new method to estimate in vivo fluorescein isothiocyanate (FITC)-labelled Lactobacillus salivarius DSPV 001P counts during passage through the gastrointestinal tract (GIT) of broilers. Forty-five, 1 d old Cobb broilers were used in this trial. Programmed necropsies were performed 30 min, 6 h, and 12 h after the administration of the probiotic bacterium, and samples of liver, crop, duodenum, caecum, and bursa of fabricius were collected. To determine the spatial and temporal transit of L. salivarius DSPV 001P in broilers, the number of bacteria as well as its respective fluorescent signal produced by FITC were measured. In order to observe the relationship between the variables, a logistic regression analysis was applied. The amount of fluorescence could be used as an indicator of fluorescent probiotic bacteria in the crop and duodenum 30 min after probiotic bacterium supplementation. In addition, the fluorescent signal could be used to estimate bacterial counts in caecum 6 and 12 h after L. salivarius DSPV 001P administration. To the best of our knowledge, this research is the first in vivo trial to employ the bacterial FITC-labelling technique in order to enumerate probiotic bacteria during gastrointestinal transit in broilers.
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Kakaiya, R. M., T. L. Kiraly, and R. G. Cable. "Concanavalin A Induces Patching/Capping of the Platelet Membrane Glycoprotein llb/llla Complex." Thrombosis and Haemostasis 59, no. 02 (1988): 281–83. http://dx.doi.org/10.1055/s-0038-1642771.
Повний текст джерелаАнотація:
SummaryTwo recent reports have shown platelet patching/capping by concanavalin A (Con A). In these studies, Con A receptors were shown to mobilize from pseudopodia and lamellipodia to the central cell parts during platelet attachment and spreading. The molecular mechanism underlying Con A receptor capping was not examined in either study.Con a binds maximally to human platelet membrane glycoproteins IIb and IIIa. In order to test whether Con A-induced capping caused the capping of this membrane glycoprotein complex, we treated normal human platelets with unlabeled Con A. After fixation, platelets were further treated with mouse monoclonal antibodies against the membrane glycoprotein IIb/IIIa complex and stained with fluorescein isothiocyanate (FITC) tagged anti-mouse IgG. An average of 16% platelets manifested capping with one monoclonal antibody preparation (N = 2) and 12% with a second preparation (N = 2).Control studies showed that only 18% of normal human fresh platelets exhibit capping with FITC-Con A (N = 17). If platelets were first incubated with unlabeled Con A, followed by staining with FITC-labeled anti-Con A antibody, an average of 15% platelets manifested caps (N = 17). Capping was inhibited by methyl-alpha-D-mannopyranoside (a known inhibitor of Con A), at cold temperature and by pre-treatment of platelets with colchicine.Our studies confirm the earlier findings on Con A induced capping. Also, our findings suggest that the molecular mechanism for Con A receptor capping involves patching and capping of the platelet membrane glycoprotein IIb/IIIa complex. It is possible that glycoprotein Ilb/IIIa redistribution might be intimately involved during platelet attachment and spreading.
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Ohata, Kenji, and Anthony Marmarou. "Clearance of brain edema and macromolecules through the cortical extracellular space." Journal of Neurosurgery 77, no. 3 (September 1992): 387–96. http://dx.doi.org/10.3171/jns.1992.77.3.0387.
Повний текст джерелаАнотація:
✓ The transit routes of fluid and particulate matter through brain tissue remain unclear. The object of this study was to examine the movement of macromolecules through brain tissue to further clarify the clearance pathways of edema proteins as they migrate toward the cortex. For this purpose, albumin solution (20 µl rat albumin diluted to 65 mg/ml with mock cerebrospinal fluid (CSF)) was intracerebrally infused into the caudate putamen, and the migration through brain tissue as well as through the ultrastructure of the cortical surfaces was explored using an immunocytochemical technique. The authors observed immunoreactive product on the glial limitans and pial lining as well as in the extracellular space of the cortical neuropil at 24 hours postinfusion, confirming that the protein had reached the cortical surface. To confirm the efflux of macromolecules into the subarachnoid CSF, 71,200 D fluorescein isothiocyanatedextran (FITC-dextran 71,200) was infused; cortical surfaces of brains removed en bloc as well as coronal sections were macroscopically observed under ultraviolet illumination at 15 minutes and 24 hours postinfusion. It was observed that infused FITC-dextran 71,200 mainly localized in the cortical white matter and caudate putamen of the infusion site at 15 minutes postinfusion and by 24 hours was distributed in the entire cortex of the infused hemisphere. However, the dynamics of lower-molecular-weight substances was completely different. The spatial distribution of FITC-dextran 4400 diverged upward toward the cortical surface and spread more extensively than FITC-dextran 71,200. These observations were consistent with a diffusion process as the spread of the tracer was dependent upon molecular size. These studies provide compelling evidence that a process other than bulk flow was involved in the spread of macromolecules through the extracellular space of the normal cortical neuropil to sink into the subarachnoid space. It was concluded that the CSF pathway via the extracellular space of the cortical neuropil is a primary route for clearance of extracellular edema proteins to the subarachnoid space and that diffusion is involved in this process.