scholarly journals HIGH-RESOLUTION RADIOAUTOGRAPHY OF PHLORIZIN-3H IN RINGS OF HAMSTER INTESTINE

1967 ◽  
Vol 35 (3) ◽  
pp. 605-618 ◽  
Author(s):  
Charles E. Stirling
Keyword(s):  
Epithelial Cell ◽  
Lamina Propria ◽  
Langmuir Adsorption Isotherm ◽  
Cell Compartment ◽  
Langmuir Adsorption ◽  
Half Saturation Constant ◽  
Cell Accumulation ◽  
Freeze Dried ◽  
Galactose Content

Quantitative light microscope radioautographs of galactose-3H and phlorizin-3H were prepared from freeze-dried plastic-embedded hamster small intestine incubated in vitro. The usual uphill epithelial cell accumulation of galactose accompanied by a somewhat smaller lamina propria accumulation was observed in control tissue incubated 3 min in 1 mM galactose-3H. The addition of 5 x 10-4 M phlorizin to the medium blocked uphill accumulation, but did not prevent galactose equilibration with the epithelial cells. The galactose content of the lamina propria was considerably less than the galactose content of the epithelial cell. Varying the phlorizin-3H content of the medium from 0.6 to 60 µM revealed a brush border binding of phlorizin which followed a Langmuir adsorption isotherm with a half-saturation constant of 13 µM and a maximum binding of 84 µmoles of phlorizin/liter of microvilli or 2.6 x 106 sites/epithelial cell. The phlorizin content of the epithelial cell compartment, excluding microvilli, never exceeded 10% that of the medium after 20 min of incubation. These findings directly support the view that phlorizin is a nontransported inhibitor which binds glucose-galactose carriers at the surface of epithelial cell microvilli.

Intestinal absorption of L-phenylalanine in vitro

1961 ◽  
Vol 200 (3) ◽  
pp. 501-504 ◽  
Author(s):  
Richard P. Spencer ◽  
A. H. Samiy
Keyword(s):  
Small Intestine ◽  
Steady State ◽  
Adsorption Isotherm ◽  
Intestinal Absorption ◽  
Concentration Gradient ◽  
Langmuir Adsorption Isotherm ◽  
Langmuir Adsorption ◽  
High Concentrations ◽  
Phenylalanine Transport

When initially present at the same concentration on both sides, net transport of L-phenylalanine by everted hamster intestinal sacs reached a steady state at 1 hour. The net amount transported, as a function of the final mucosal concentration, was described by a Langmuir adsorption isotherm. An equation was derived showing that the concentration gradient also obeyed such a formulation. Sacs from the midintestine transported more L-phenylalanine than proximal and distal end sacs. Lineweaver-Burk plots of the concentration gradient against final mucosal concentrations showed the process in end sacs to have the same KM (1.8 x 10–3 M) as that in midintestinal sacs; those from midgut, however, had a greater capacity. L-Phenylalanine absorption likely occurs by different amounts of the same mechanism throughout the small intestine. High concentrations of L-phenylalanine resulted in a depression of net transport. The importance of the concentration gradient in revealing the fall in net transport in this and other systems was stressed. l-Phenylalanine transport was also depressed by L-tryptophan and L-methionine.

scholarly journals HIGH-RESOLUTION RADIOAUTOGRAPHY OF GALACTOSE-3H ACCUMULATION IN RINGS OF HAMSTER INTESTINE

1967 ◽  
Vol 35 (3) ◽  
pp. 585-604 ◽  
Author(s):  
Charles E. Stirling ◽  
William B. Kinter
Keyword(s):  
Lamina Propria ◽  
Brush Border ◽  
Technical Problem ◽  
Water Soluble ◽  
Cell Cytoplasm ◽  
The Past ◽  
Columnar Cells ◽  
Transcellular Diffusion ◽  
Galactose Content

Radioautography of water-soluble substances has posed a major technical problem for the past decade. Utilizing silicone-impregnated plastic sections of frozen-dried tissue, a quantitative method was developed for studying distribution of 3H-labeled galactose, mannitol, and phlorizin. The content of a 2-µ band may be measured with an accuracy of ±20% by light microscopy; radioautographs may also be prepared for the electron microscope. Results with intestinal tissue incubated 1–10 min in vitro and, then, frozen rapidly indicate that the first step in galactose absorption is uphill transport into the brush border of the columnar epithelium. Correction of galactose content for the mannitol space in the brush border suggests that the sugar pump is located at the surface of the microvilli. Further evidence for the surface locus of the glucose-galactose pump was obtained with phlorizin (next paper, reference 40). The galactose content of columnar cell cytoplasm always equalled that of microvilli and no transcellular diffusion gradient could be detected; during the first minutes of incubation, however, a gradient did exist between nucleoplasm and cytoplasm. Downhill exit of galactose from columnar cells may have proceeded either directly across basal membranes to adjacent lamina propria or indirectly via open intercellular spaces. Lastly, even in the absence of muscularis, the connective tissue of the lamina propria constituted enough of a diffusion barrier so that it served as a secondary accumulating compartment for galactose under present in vitro conditions.

RANTES mediates TNF-dependent lamina propria mast cell accumulation and barrier dysfunction in neurogenic cystitis

2007 ◽  
Vol 292 (5) ◽  
pp. F1372-F1379 ◽  
Author(s):  
Michael C. Chen ◽  
Pavitra Keshavan ◽  
Greg D. Gregory ◽  
David J. Klumpp
Keyword(s):  
Mast Cell ◽  
Lamina Propria ◽  
Pseudorabies Virus ◽  
Cell Trafficking ◽  
Barrier Dysfunction ◽  
Patient Morbidity ◽  
Cell Accumulation ◽  
Factor Α

Barrier dysfunction of the urinary bladder is postulated to contribute to patient morbidity in the bladder inflammatory disease interstitial cystitis (IC). IC is often considered a neurogenic cystitis, but the mechanisms underlying barrier dysfunction are unclear. In murine neurogenic cystitis induced by pseudorabies virus (PRV), we previously observed formation of urothelial lesions characterized by urothelial apoptosis and urothelial discontinuities. Lesion formation was preceded by mast cell trafficking to the lamina propria, and trafficking was mediated by tumor necrosis factor-α (TNF). Here, we found that supernatants of TNF-treated urothelial cultures promoted chemotaxis of bone marrow-derived mast cells in vitro that was blocked by anti-RANTES antibodies but unaffected by anti-TNF antibodies. In vivo, PRV infection of wild-type mice induced RANTES expression in the urothelium that was temporally coincident with lamina propria mast cell accumulation (maximum at days 3–4 following infection) and was not induced in TNF−/− mice, TNFR1/2−/− mice, or mice treated with anti-TNF antibodies. Anti-RANTES antibodies blocked PRV-induced lamina propria mast cell accumulation 56% and reduced the prevalence of animals with detectable lesions 42%, relative to isotype control antibodies. Bladder barrier function was quantified by measuring transepithelial resistance (TER). PRV induced a 49% loss of TER in the presence of control antibodies, but mice treated with anti-RANTES antibodies exhibited reduced TER loss (16%, P < 0.01). These data demonstrate that RANTES plays a key role in the pathogenesis of neurogenic cystitis and suggest that chemokines may represent novel therapeutic targets for IC patients with mast cell-associated disease.

Preparation of cultured astrocytes for x-ray microanalysis

1989 ◽  
Vol 47 ◽  
pp. 988-989
Author(s):  
N.K.R. Smith ◽  
K.E. Hunter ◽  
P. Mobley ◽  
L.P. Felpel
Keyword(s):  
Cultured Cells ◽  
Elemental Content ◽  
Elemental Distribution ◽  
Sprague Dawley ◽  
X Ray ◽  
Cultured Astrocytes ◽  
Freeze Dried ◽  
Ultimate Objective

Electron probe energy dispersive x-ray microanalysis (XRMA) offers a powerful tool for the determination of intracellular elemental content of biological tissue. However, preparation of the tissue specimen , particularly excitable central nervous system (CNS) tissue , for XRMA is rather difficult, as dissection of a sample from the intact organism frequently results in artefacts in elemental distribution. To circumvent the problems inherent in the in vivo preparation, we turned to an in vitro preparation of astrocytes grown in tissue culture. However, preparations of in vitro samples offer a new and unique set of problems. Generally, cultured cells, growing in monolayer, must be harvested by either mechanical or enzymatic procedures, resulting in variable degrees of damage to the cells and compromised intracel1ular elemental distribution. The ultimate objective is to process and analyze unperturbed cells. With the objective of sparing others from some of the same efforts, we are reporting the considerable difficulties we have encountered in attempting to prepare astrocytes for XRMA.Tissue cultures of astrocytes from newborn C57 mice or Sprague Dawley rats were prepared and cultured by standard techniques, usually in T25 flasks, except as noted differently on Cytodex beads or on gelatin. After different preparative procedures, all samples were frozen on brass pins in liquid propane, stored in liquid nitrogen, cryosectioned (0.1 μm), freeze dried, and microanalyzed as previously reported.

Freeze-Dried Clopidogrel Loaded Lyotropic Liquid Crystal: Box-Behnken Optimization, In-Vitro and In-Vivo Evaluation

2020 ◽  
Vol 17 (3) ◽  
pp. 207-217
Author(s):  
Eman A. Hakeem ◽  
Galal M. El-Mahrouk ◽  
Ghada Abdelbary ◽  
Mahmoud H. Teaima
Keyword(s):  
Statistical Analysis ◽  
Oral Bioavailability ◽  
Quadratic Model ◽  
Lipid Phase ◽  
Individual Response ◽  
In Vivo Evaluation ◽  
Freeze Dried ◽  
Suggested Formula

Background: Clopidogrel (CLP) suffers from extensive first pass metabolism results in a negative impact on its oral systemic bioavailability. Cubosomes are Lyotropic Liquid Crystalline (LLC) nano-systems comprising monoolein, a steric stabilizer and an aqueous system, it considered a promising carrier for different pharmaceutical compounds. Box-Behnken Design (BBD) is an efficient tool for process analysis and optimization skipping forceful treatment combinations. Objective: The study was designed to develop freeze-dried clopidogrel loaded LLC (cubosomes) for enhancement of its oral bioavailability. Methods: A 33 BBD was adopted, the studied independent factors were glyceryl monooleate (GMO lipid phase), Pluronic F127 (PL F127steric stabilizer) and polyvinyl alcohol powder (stabilizer). Particle Size (PS), Polydispersity Index (PDI) and Zeta Potential (ZP) were set as independent response variables. Seventeen formulae were prepared in accordance with the bottom up approach and in-vitro evaluated regarding PS, PDI and ZP. Statistical analysis and optimization were achieved using design expert software®, then the optimum suggested formula was prepared, in-vitro revaluated, freeze-dried with 3% mannitol (cryoprotectant), solid state characterized and finally packed in hard gelatin capsule for comparative in-vitro release and in-vivo evaluation to Plavix®. Results: Results of statistical analysis of each individual response revealed a quadratic model for PS and PDI where a linear model for ZP. The optimum suggested formula with desirability factor equal 0.990 consisting of (200 mg GMO, 78.15 mg PL F127 and 2% PVA). LC/MS/MS study confirmed significant higher C>max, AUC>0-24h and AUC>0-∞ than that of Plavix®. Conclusion: The results confirm the capability of developed carrier to overcome the low oral bioavailability.

Met -/- kidneys express epithelial cells that chemotax and form tubules in response to EGF receptor ligands

1997 ◽  
Vol 272 (2) ◽  
pp. F222-F228
Author(s):  
C. Kjelsberg ◽  
H. Sakurai ◽  
K. Spokes ◽  
C. Birchmeier ◽  
I. Drummond ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epithelial Cell ◽  
Egf Receptor ◽  
Transforming Growth Factor ◽  
Growth Factor Receptor ◽  
Receptor Ligands ◽  
Tubule Formation ◽  
Human Papillomavirus 16 ◽  
Immortalized Cells

The growth factor/receptor combination of hepatocyte growth factor (HGF)/c-met has been postulated to be critical for mesenchymal-to-epithelial conversion and tubule formation in the developing kidney. We therefore isolated and immortalized cells from embryonic kidneys of met -/- transgenic mice to determine whether these cells were epithelial and able to chemotax and form tubules in vitro. The cells were immortalized with retrovirus expressing human papillomavirus 16 (HPV 16) E6/E7 genes. Two rapidly dividing clones were isolated and found to express the epithelial cell markers cytokeratin, zonula occludens-1, and E-cadherin but not to express the fibroblast marker vimentin. The met -/- cells were able to chemotax in response to epidermal growth factor and transforming growth factor-alpha (TGF-alpha) and form tubules in vitro in response to TGF-alpha but not HGF. These experiments suggest that the HGF/c-met axis is not essential for epithelial cell development in the embryonic kidney and demonstrate that other growth factors are capable of supporting early tubulogenesis.

An in vitro senescence model of gingival epithelial cell induced by hydrogen peroxide treatment

Odontology ◽  
2021 ◽  
Author(s):  
Sarita Giri ◽  
Ayuko Takada ◽  
Durga Paudel ◽  
Koki Yoshida ◽  
Masae Furukawa ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Epithelial Cell ◽  
Gingival Epithelial Cell ◽  
Hydrogen Peroxide Treatment

scholarly journals In vitro co-culture of human intestinal organoids and lamina propria-derived CD4+ T cells

2021 ◽  
Vol 2 (2) ◽  
pp. 100519
Author(s):  
Renée R.C.E. Schreurs ◽  
Martin E. Baumdick ◽  
Agata Drewniak ◽  
Madeleine J. Bunders
Keyword(s):  
T Cells ◽  
Lamina Propria ◽  
Cd4 T Cells ◽  
Intestinal Organoids

scholarly journals A High-Throughput Metabolic Microarray Assay Reveals Antibacterial Effects of Black and Red Raspberries and Blackberries against Helicobacter pylori Infection

Antibiotics ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 845
Author(s):  
Candace Goodman ◽  
Katrina N. Lyon ◽  
Aitana Scotto ◽  
Cyra Smith ◽  
Thomas A. Sebrell ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
High Throughput ◽  
Antibacterial Properties ◽  
Treatment Strategies ◽  
Helicobacter Pylori Infection ◽  
Antibacterial Effects ◽  
Biolog System ◽  
Freeze Dried ◽  
H Pylori

Helicobacter pylori infection is commonly treated with a combination of antibiotics and proton pump inhibitors. However, since H. pylori is becoming increasingly resistant to standard antibiotic regimens, novel treatment strategies are needed. Previous studies have demonstrated that black and red berries may have antibacterial properties. Therefore, we analyzed the antibacterial effects of black and red raspberries and blackberries on H. pylori. Freeze-dried powders and organic extracts from black and red raspberries and blackberries were prepared, and high-performance liquid chromatography was used to measure the concentrations of anthocyanins, which are considered the major active ingredients. To monitor antibiotic effects of the berry preparations on H. pylori, a high-throughput metabolic growth assay based on the Biolog system was developed and validated with the antibiotic metronidazole. Biocompatibility was analyzed using human gastric organoids. All berry preparations tested had significant bactericidal effects in vitro, with MIC90 values ranging from 0.49 to 4.17%. Antimicrobial activity was higher for extracts than powders and appeared to be independent of the anthocyanin concentration. Importantly, human gastric epithelial cell viability was not negatively impacted by black raspberry extract applied at the concentration required for complete bacterial growth inhibition. Our data suggest that black and red raspberry and blackberry extracts may have potential applications in the treatment and prevention of H. pylori infection but differ widely in their MICs. Moreover, we demonstrate that the Biolog metabolic assay is suitable for high-throughput antimicrobial susceptibility screening of H. pylori.

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