Coconut milk regulates the allergic response in Balb/c mice sensitized with whey proteins

2018 ◽  
Vol 8 (1) ◽  
pp. 19-25
Author(s):  
Yamina Benaissa ◽  
Samia Addou ◽  
Wafaa Dib ◽  
Hadria Grar ◽  
Omar Kheroua ◽  
...  
Keyword(s):  
Clinical Symptoms ◽  
Ex Vivo ◽  
Short Circuit ◽  
Ussing Chamber ◽  
Whey Proteins ◽  
Milk Proteins ◽  
Short Circuit Current ◽  
Coconut Milk ◽  
Intestinal Damage

In this work the effect of coconut milk on mice sensitized to cow's milk proteins was evaluated. Balb/c mice treated orally for twenty eight days with coconut milk were sensitized intraperitoneally with β-lactoglobulin (β-Lg) or α-Lactoglobulin (α-Lac). We used Ussing chamber to analyze ex vivo electrical parameters characterizing the intestinal tissue of mice by measuring the variations of the short current circuit Isc (μA/cm2) as well as the epithelial conductance (G). Jejunal fragments of sensitized and treated mice were mounted in Ussing chamber and stimulated by the deposit of β-Lg or α-Lac. Symptom scores were determined after in vivo challenge to β-Lg or α-Lac. Intestinal damage was assessed by histological analysis. Coconut milk influ-enced the electrophysiological parameters by significantly decreasing the short-circuit current (Isc) (p < 0.001) and the epithelial conductance ((p< 0.01 and (p < 0.001, respectively). Moreover, in coconut milk-treated mice, no significant clinical symptoms were observed. Analysis of histological sections revealed that coconut milk reduced the microscopic lesions induced by β-Lg or α-Lac sensitization. We speculate that the administration of coconut milk could prevent the systemic and anaphylactic responses in sensitized mice.

scholarly journals Assessment of Distinct Electrophysiological Parameters in Rectal Biopsies for the Choice of the Best Diagnosis/Prognosis Biomarkers for Cystic Fibrosis

2020 ◽  
Vol 11 ◽  
Author(s):  
Iris A. L. Silva ◽  
Aires Duarte ◽  
Fernando A. L. Marson ◽  
Raquel Centeio ◽  
Tereza Doušová ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Clinical Features ◽  
Ex Vivo ◽  
Short Circuit ◽  
Ussing Chamber ◽  
Short Circuit Current ◽  
Predictive Biomarker ◽  
Sweat Chloride ◽  
Best Parameter ◽  
Rectal Biopsies

Most cases of Cystic Fibrosis (CF) are diagnosed early in life. However, people with atypical CF forms pose diagnosis dilemmas, requiring laboratory support for diagnosis confirmation/exclusion. Ex vivo analysis of fresh rectal biopsies by Ussing chamber has been the best discriminant biomarker for CF diagnosis/prognosis so far. Here we aimed to evaluate different electrophysiological parameters from Ussing chamber analysis of rectal biopsies from people with CF (PwCF) to establish the one with highest correlations with clinical features as the best CF diagnosis/prognosis biomarker. We analyzed measurements of CFTR-mediated Cl– secretion in rectal biopsies from 143 individuals (∼592 biopsies), the largest cohort so far analyzed by this approach. New parameters were analyzed and compared with the previous biomarker, i.e., the IBMX (I)/Forskolin (F)/Carbachol (C)-stimulated short-circuit current (I’sc–I/F/C). Correlations with clinical features showed that the best parameter corresponded to voltage measurements of the I/F + (I/F/CCH) response (VI/F+I/F/C), with higher correlations vs. I’sc–I/F/C for: sweat chloride (59 vs. 52%), fecal elastase (69 vs. 55%) and lung function, measured by FEV1 (27 vs. 20%). Altogether data show that VI/F+I/F/C is the most sensitive, reproducible, and robust predictive biomarker for CF diagnosis/prognosis effectively discriminating classical, atypical CF and non-CF groups.

scholarly journals Involvement of the EnteroaggregativeEscherichia coli Plasmid-Encoded Toxin in Causing Human Intestinal Damage

1999 ◽  
Vol 67 (10) ◽  
pp. 5338-5344 ◽  
Author(s):  
Ian R. Henderson ◽  
Susan Hicks ◽  
Fernando Navarro-Garcia ◽  
Waldir P. Elias ◽  
Alan D. Philips ◽  
...  
Keyword(s):  
Short Circuit ◽  
Ussing Chamber ◽  
Short Circuit Current ◽  
Mucosal Damage ◽  
Intestinal Damage ◽  
Intestinal Epithelial ◽  
T84 Cells ◽  
Aperture Diameter ◽  
Eaec Strain

ABSTRACT Enteroaggregative Escherichia coli (EAEC) strains have been shown to adhere to human intestinal tissue in an in vitro organ culture (IVOC) model, and certain strains manifest mucosal toxicity. We have recently described the EAEC plasmid-encoded toxin (Pet), a member of a specific serine protease subclass of the autotransporter proteins. When injected into rat ileal loops, Pet both elicited fluid accumulation and had cytotoxic effects on the mucosa. Furthermore, the Pet protein caused rises in short circuit current from rat jejunal tissue mounted in a Ussing chamber and rounding of intestinal epithelial cells in culture. We therefore hypothesized that the mucosal pathology induced by EAEC strains in the IVOC model was related to expression of the Pet protein. Here, we have examined the effects of EAEC strain 042 and its isogenic pet mutant in the IVOC model. 042-infected colonic explants exhibited dilation of crypt openings, increased cell rounding, development of prominent intercrypt crevices, and absence of apical mucus plugs. Colonic tissue incubated with the pet mutant exhibited significantly fewer mucosal abnormalities both subjectively and as quantitated morphometrically by measurement of crypt aperture diameter. Mucosal effects were restored upon complementation of the pet mutation intrans. Interestingly, we found that the ability of 042 to damage T84 cells was not dependent upon Pet. The data suggest that the Pet toxin is active on the human intestinal mucosa but that EAEC may have other mechanisms of eliciting mucosal damage.

Tissue- and Salinity-specific Na–Cl Cotransporter (NCC) Orthologues Involved in the Sea lamprey (Petromyzon Marinus) Adaptive Osmoregulation

2021 ◽  
Author(s):  
A. Barany ◽  
C. A. Shaughnessy ◽  
R. M. Pelis ◽  
J. Fuentes ◽  
J. M. Mancera ◽  
...  
Keyword(s):  
Water Absorption ◽  
Ex Vivo ◽  
Short Circuit ◽  
Ussing Chamber ◽  
Petromyzon Marinus ◽  
Short Circuit Current ◽  
Sea Lamprey ◽  
Intestinal Tissue ◽  
Distal Intestine ◽  
Phylogenetic Comparison

Abstract Two ncc orthologues (termed ncca and nccb) were found in the sea lamprey genome, whereas nkcc2 was not. In a phylogenetic comparison among other vertebrate amino acids, NCC and NKCC deduced sequences, the sea lamprey NCC’s occupied basal positions within the NCC clade. In freshwater, ncca mRNA was found only in the gill and nccb only in the intestine, whereas both were found in the kidney. Acclimation to seawater increased nccb mRNA in the intestine and kidney. Intestinal nccb mRNA also increased during late metamorphosis. The electrophysiological approach in the Ussing chamber of intestinal tissue ex vivo showed significant differences between freshwater and seawater-acclimated juveniles. Luminal application of indapamide (NCC inhibitor) resulted in 73 and 30% inhibition of short-circuit current (Isc) in the proximal and distal intestine, respectively. The luminal application of bumetanide (NKCC inhibitor) did not affect intestinal Isc. Indapamide also inhibited ex vivo intestinal water absorption. Our results indicate that NCCb is likely the key passive ion cotransporter protein for ion uptake by the lamprey intestine to facilitate water absorption in seawater. As such, the preparatory increases in intestinal nccb mRNA expression during metamorphosis are likely critical to the development of whole animal salinity tolerance.

Mechanisms of disposal of acid and alkali in rabbit duodenum

1975 ◽  
Vol 229 (6) ◽  
pp. 1641-1648 ◽  
Author(s):  
RG Fiddian-Green ◽  
W Silen
Keyword(s):  
Short Circuit ◽  
Ussing Chamber ◽  
Short Circuit Current ◽  
Duodenal Mucosa ◽  
Ringer Solution ◽  
Co2 Production ◽  
Rapid Phase ◽  
Ussing Chambers

Stripped duodenal mucosa of rabbits was mounted in Ussing chambers containing a Ringer solution gassed with 100% O2. The disappearance of acid or alkali from the mucosal solution of short-circuited tissue was measured with a pH stat while the serosal pH was kept at 7.4. The duodenum rapidly disposed of both acid and alkali; neither property was altered by gassing with N2 while iodoacetate was in the perfusing solutions. Prevention of release of CO2 from the mucosal chamber obliterated the early rapid phase of acid disposal by the mucosa while a similar maneuver in the serosal chamber increased the appearance of serosal acid without altering the rate of acid disposal. Gut sacs of rabbit duodenum in vitro and in vivo showed a positive correlation between acid disposal and the rate of luminal CO2 production. While acid disposal progressively decreased with time for the in vitro gut sacs, the in vivo gut sac showed no fatigue in this respect. Luminal acidification in the Ussing chamber was associated with a profound reduction in short-circuit current (Isc), partially reversible by elevation of the mucosal pH but not by luminal glucose. Our data suggest that acid disposal occurs in part by intraluminal neutralization and in part by diffusion into the mucosa.

scholarly journals Enterotoxigenicity of Mature 45-Kilodalton and Processed 35-Kilodalton Forms of Hemagglutinin Protease Purified from a Cholera Toxin Gene-Negative Vibrio cholerae Non-O1, Non-O139 Strain

2006 ◽  
Vol 74 (5) ◽  
pp. 2937-2946 ◽  
Author(s):  
A. Ghosh ◽  
D. R. Saha ◽  
K. M. Hoque ◽  
M. Asakuna ◽  
S. Yamasaki ◽  
...  
Keyword(s):  
Cholera Toxin ◽  
Vibrio Cholerae ◽  
Hela Cells ◽  
Histopathological Examination ◽  
Short Circuit ◽  
Ussing Chamber ◽  
Short Circuit Current ◽  
Toxin Gene ◽  
Dependent Manner ◽  
A Cell

ABSTRACT Cholera toxin gene-negative Vibrio cholerae non-O1, non-O139 strain PL-21 is the etiologic agent of cholera-like syndrome. Hemagglutinin protease (HAP) is one of the major secretory proteins of PL-21. The mature 45-kDa and processed 35-kDa forms of HAP were purified in the presence and absence of EDTA from culture supernatants of PL-21. Enterotoxigenicities of both forms of HAP were tested in rabbit ileal loop (RIL), Ussing chamber, and tissue culture assays. The 35-kDa HAP showed hemorrhagic fluid response in a dose-dependent manner in the RIL assay. Histopathological examination of 20 μg of purified protease-treated rabbit ileum showed the presence of erythrocytes and neutrophils in the upper part of the villous lamina propria. Treatment with 40 μg of protease resulted in gross damage of the villous epithelium with inflammation, hemorrhage, and necrosis. The 35-kDa form of HAP, when added to the lumenal surface of rat ileum loaded in an Ussing chamber, showed a decrease in the intestinal short-circuit current and a cell rounding effect on HeLa cells. The mature 45-kDa form of HAP showed an increase in intestinal short-circuit current in an Ussing chamber and a cell distending effect on HeLa cells. These results show that HAP may play a role in the pathogenesis of PL-21.

Basolateral Cl− uptake mechanisms in Xenopus laevis lung epithelium

2010 ◽  
Vol 299 (1) ◽  
pp. R92-R100 ◽  
Author(s):  
Jens Berger ◽  
Martin Hardt ◽  
Wolfgang G. Clauss ◽  
Martin Fronius
Keyword(s):  
Xenopus Laevis ◽  
Ion Transport ◽  
Anion Exchanger ◽  
Short Circuit ◽  
Ussing Chamber ◽  
Short Circuit Current ◽  
Transport Processes ◽  
Lung Epithelium ◽  
Active Ion Transport ◽  
Uptake Mechanisms

A thin liquid layer covers the lungs of air-breathing vertebrates. Active ion transport processes via the pulmonary epithelial cells regulate the maintenance of this layer. This study focuses on basolateral Cl− uptake mechanisms in native lungs of Xenopus laevis and the involvement of the Na+/K+/2 Cl− cotransporter (NKCC) and HCO3−/Cl− anion exchanger (AE), in particular. Western blot analysis and immunofluorescence staining revealed the expression of the NKCC protein in the Xenopus lung. Ussing chamber experiments demonstrated that the NKCC inhibitors (bumetanide and furosemide) were ineffective at blocking the cotransporter under basal conditions, as well as under pharmacologically stimulated Cl−-secreting conditions (forskolin and chlorzoxazone application). However, functional evidence for the NKCC was detected by generating a transepithelial Cl− gradient. Further, we were interested in the involvement of the HCO3−/Cl− anion exchanger to transepithelial ion transport processes. Basolateral application of DIDS, an inhibitor of the AE, resulted in a significantly decreased the short-circuit current (ISC). The effect of DIDS was diminished by acetazolamide and reduced by increased external HCO3− concentrations. Cl− secretion induced by forskolin was decreased by DIDS, but this effect was abolished in the presence of HCO3−. These experiments indicate that the AE at least partially contributes to Cl− secretion. Taken together, our data show that in Xenopus lung epithelia, the AE, rather than the NKCC, is involved in basolateral Cl− uptake, which contrasts with the common model for Cl− secretion in pulmonary epithelia.

Metabolic Support of Chloride-dependent Short-circuit Current Across Locust Rectum

1982 ◽  
Vol 99 (1) ◽  
pp. 349-362
Author(s):  
M. CHAMBERLIN ◽  
J. E. PHILLIPS
Keyword(s):  
Cyclic Amp ◽  
Short Circuit ◽  
Malpighian Tubule ◽  
Short Circuit Current ◽  
Metabolic Substrates ◽  
Endogenous Substrates ◽  
Tubule Fluid ◽  
Substrate Source ◽  
Stimulation Of

1. Recta of desert locusts were short-circuited and depleted of endogenous substrates by exposing them to saline containing cyclic AMP but no metabolites. Individual substrates were then added to substrate-depleted recta and the change in short-circuit current (Isc) monitored. 2. Proline or glucose (50 mM) caused by far the largest increase in Isc of all substrates tested. Stimulation of the Isc by proline was not dependent upon external sodium, but did require external chloride. 3. Physiological levels of proline also caused a large increase in Isc, while physiological levels of glucose produced a much smaller stimulation. Over 90% of the proline-dependent Isc stimulation can be produced by adding 15 mM proline solely to the lumen side of the tissue. 4. These results are discussed with regard to rectal oxidative metabolism and availability of metabolic substrates in vivo. High levels of proline in Malpighian tubule fluid are probably the major substrate source for rectal Cl−transport. Note:

Active transport of magnesium across the isolated midgut of Hyalophora cecropia

1975 ◽  
Vol 63 (2) ◽  
pp. 313-320
Author(s):  
J. L. Wood ◽  
A. M. Jungreis ◽  
W. R. Harvey
Keyword(s):  
Steady State ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Potassium Transport ◽  
Magnesium Concentration ◽  
Magnesium Transport ◽  
Wet Weight ◽  
Net Flux ◽  
Burk Plot

1. The 28Mg-measured net flux of magnesium from lumen-side to haemolymph-side of the isolated and short-circuited midgut was 1.97 +/− 0.28 mu-equiv cm(−2) /(−1) in 8 mM-Mg2+. 2. The magnesium-influx shows a delay before the tracer steady-state is attained, indicating the existence of a magnesium-transport pool equivalent to 6.7 mu-equiv/g wet weight of midgut tissue. 3. Magnesium depresses the short-circuit current produced the midgut but not the potassium transport, the depression being equal to the rate of magnesium transport. 4. Magnesium transport yields a linear Lineweaver-Burk plot with an apparent Km of 34 mM-Mg2+ and an apparent Vmax of 14.9 mu-equiv cm(−1) /(−1). 5. Magnesium is actively transported across the midgut and contributes to the regulation of the haemolymph magnesium concentration in vivo.

scholarly journals Enterohemorrhagic E. coli (EHEC)—Secreted Serine Protease EspP Stimulates Electrogenic Ion Transport in Human Colonoid Monolayers

Toxins ◽  
2018 ◽  
Vol 10 (9) ◽  
pp. 351 ◽  
Author(s):  
C. Tse ◽  
Julie In ◽  
Jianyi Yin ◽  
Mark Donowitz ◽  
Michele Doucet ◽  
...  
Keyword(s):  
Ion Transport ◽  
Serine Protease ◽  
Shiga Toxin ◽  
Short Circuit ◽  
Ussing Chamber ◽  
Adult Stem Cell ◽  
Short Circuit Current ◽  
Watery Diarrhea ◽  
E Coli ◽  
Electrogenic Ion Transport

One of the characteristic manifestations of Shiga-toxin-producing Escherichia coli (E. coli) infection in humans, including EHEC and Enteroaggregative E. coli O104:H4, is watery diarrhea. However, neither Shiga toxin nor numerous components of the type-3 secretion system have been found to independently elicit fluid secretion. We used the adult stem-cell-derived human colonoid monolayers (HCM) to test whether EHEC-secreted extracellular serine protease P (EspP), a member of the serine protease family broadly expressed by diarrheagenic E. coli can act as an enterotoxin. We applied the Ussing chamber/voltage clamp technique to determine whether EspP stimulates electrogenic ion transport indicated by a change in short-circuit current (Isc). EspP stimulates Isc in HCM. The EspP-stimulated Isc does not require protease activity, is not cystic fibrosis transmembrane conductance regulator (CFTR)-mediated, but is partially Ca2+-dependent. EspP neutralization with a specific antibody reduces its potency in stimulating Isc. Serine Protease A, secreted by Enteroaggregative E. coli, also stimulates Isc in HCM, but this current is CFTR-dependent. In conclusion, EspP stimulates colonic CFTR-independent active ion transport and may be involved in the pathophysiology of EHEC diarrhea. Serine protease toxins from E. coli pathogens appear to serve as enterotoxins, potentially significantly contributing to watery diarrhea.

Calcium secretion in canine tracheal mucosa

1985 ◽  
Vol 59 (4) ◽  
pp. 1191-1195 ◽  
Author(s):  
F. J. Al-Bazzaz ◽  
T. Jayaram
Keyword(s):  
Short Circuit ◽  
Short Circuit Current ◽  
Open Circuit ◽  
Potential Difference ◽  
Secretory Process ◽  
Metabolic Inhibitor ◽  
Tracheal Mucosa ◽  
Transepithelial Potential ◽  
Transepithelial Potential Difference

Calcium (Ca) affects many cellular functions of the respiratory tract mucosa and might alter the viscoelastic properties of mucus. To evaluate Ca homeostasis in a respiratory epithelium we investigated transport of Ca by the canine tracheal mucosa. Mucosal tissues were mounted in Ussing-type chambers and bathed with Krebs-Henseleit solution at 37 degrees C. Unidirectional fluxes of 45Ca were determined in tissues that were matched by conductance and short-circuit current (SCC). Under short-circuit conditions there was a significant net Ca secretion of 1.82 +/- 0.36 neq . cm-2 . h-1 (mean +/- SE). Under open-circuit conditions, where the spontaneous transepithelial potential difference could attract Ca toward the lumen, net Ca secretion increased significantly to 4.40 +/- 1.14 compared with 1.54 +/- 1.17 neq . cm-2 . h-1 when the preparation was short-circuited. Addition of a metabolic inhibitor, 2,4-dinitrophenol (2 mM in the mucosal bath), decreased tissue conductance and SCC and slightly decreased the unidirectional movement of Ca from submucosa to lumen. Submucosal epinephrine (10 microM) significantly enhanced Ca secretion by 2.0 +/- 0.63 neq . cm-2 . h-1. Submucosal ouabain (0.1 mM) failed to inhibit Ca secretion. The data suggest that canine tracheal mucosa secretes Ca; this secretory process is augmented by epinephrine or by the presence of a transepithelial potential difference as found under in vivo conditions.

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