In situ characterization of carbonic anhydrase activity in isolated rat lungs

1990 ◽  
Vol 69 (6) ◽  
pp. 2155-2162 ◽  
Author(s):  
T. A. Heming ◽  
A. Bidani

Lung carbonic anhydrase (CA) participates directly in plasma CO2-HCO3(-)-H+ reactions. To characterize pulmonary CA activity in situ, CO2 excretion and capillary pH equilibration were examined in isolated saline-perfused rat lungs. Isolated lungs were perfused at 25, 30, and 37 degrees C with solutions containing various concentrations of HCO3- and a CA inhibitor, acetazolamide (ACTZ). Total CO2 excretion was partitioned into those fractions attributable to dissolved CO2, uncatalyzed HCO3- dehydration, and catalyzed HCO3- dehydration. Approximately 60% of the total CO2 excretion at each temperature was attributable to CA-catalyzed HCO3- dehydration. Inhibition of pulmonary CA diminished CO2 excretion and produced significant postcapillary perfusate pH disequilibria, the magnitude and time course of which were dependent on temperature and the extent of CA inhibition. The half time for pH equilibration increased from approximately 5 s at 37 degrees C to 14 s at 25 degrees C. For the HCO3- dehydration reaction, pulmonary CA in situ displayed an apparent inhibition constant for ACTZ of 0.9-2.2 microM, a Michaelis-Menten constant of 90 mM, a maximal reaction velocity of 9 mM/s, and an apparent activation energy of 3.0 kcal/mol.

1986 ◽  
Vol 61 (5) ◽  
pp. 1849-1856 ◽  
Author(s):  
T. A. Heming ◽  
C. Geers ◽  
G. Gros ◽  
A. Bidani ◽  
E. D. Crandall

Effects of macromolecular Prontosil-dextran inhibitors (PD) on carbonic anhydrase (CA) activity in isolated rat lungs were studied. Isolated lungs were perfused with Krebs-Ringer bicarbonate (KRB) solutions containing no inhibitor, PD 100,000 (mol wt 100,000), PD 5,000 (mol wt 5,000), or low-molecular-weight inhibitors (Prontosil or acetazolamide). The time course of effluent perfusate pH equilibration was measured in a stop-flow pH electrode apparatus. Pulmonary CO2 excretion (Vco2) was monitored by continuously recording expired CO2 concentration. The lungs were ventilated with room air and perfused at 37 degrees C with KRB prebubbled with 5% CO2- 20% O2- 75% N2. The results obtained show that both the low-molecular-weight inhibitors and PD′s caused postcapillary pH disequilibria (delta pH) in effluent perfusate. However, only acetazolamide and Prontosil caused a reduction in Vco2. These results suggest that there is an intravascular CA, presumably associated with endothelial cell membranes, that is accessible to all inhibitors used and is responsible in part for equilibration of the CO2- HCO3- -H+ reactions in the perfusate but, under the conditions used, does not affect CO2 excretion; and there is an extravascular (possibly intracellular) CA that can be inhibited by low-molecular-weight inhibitors, is primarily responsible for enhanced CO2 transfer across the alveolar-capillary barrier (perhaps via facilitation of CO2 diffusion), and is in part responsible for pH equilibration.


Reproduction ◽  
2000 ◽  
pp. 325-335 ◽  
Author(s):  
A Calvo ◽  
LM Pastor ◽  
S Bonet ◽  
E Pinart ◽  
M Ventura

Lectin histochemistry was used to perform in situ characterization of the glycoconjugates present in boar testis and epididymis. Thirteen horseradish peroxidase- or digoxigenin-labelled lectins were used in samples obtained from healthy fertile boars. The acrosomes of the spermatids were stained intensely by lectins with affinity for galactose and N-acetyl-galactosamine residues, these being soybean, peanut and Ricinus communis agglutinins. Sertoli cells were stained selectively by Maackia ammurensis agglutinin. The lamina propria of seminiferous tubules showed the most intense staining with fucose-binding lectins. The Golgi area and the apical part of the principal cells of the epididymis were stained intensely with many lectins and their distribution was similar in the three zones of the epididymis. On the basis of lectin affinity, both testis and epididymis appear to have N- and O-linked glycoconjugates. Spermatozoa from different epididymal regions showed different expression of terminal galactose and N-acetyl-galactosamine. Sialic acid (specifically alpha2,3 neuraminic-5 acid) was probably incorporated into spermatozoa along the extratesticular ducts. These findings indicate that the development and maturation of boar spermatozoa are accompanied by changes in glycoconjugates. As some lectins stain cellular or extracellular compartments specifically, these lectins could be useful markers in histopathological evaluation of diseases of boar testis and epididymis.


1983 ◽  
Author(s):  
K. Arulanandan ◽  
Y. Dafalias ◽  
L. R. Herrmann ◽  
A. Anandarajah ◽  
N. Meegoda

2016 ◽  
Vol 108 (21) ◽  
pp. 211902 ◽  
Author(s):  
Xian Chen ◽  
Nobumichi Tamura ◽  
Alastair MacDowell ◽  
Richard D. James

ACS Catalysis ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 1464-1484 ◽  
Author(s):  
Yong Han ◽  
Hui Zhang ◽  
Yi Yu ◽  
Zhi Liu

2009 ◽  
Vol 11 (20) ◽  
pp. 3958 ◽  
Author(s):  
Daniel Irimia ◽  
Rob Kortekaas ◽  
Maurice H. M. Janssen

2013 ◽  
Vol 22 ◽  
pp. 85-93
Author(s):  
Shuang Yi Liu ◽  
Min Min Tang ◽  
Ai Kah Soh ◽  
Liang Hong

In-situ characterization of the mechanical behavior of geckos spatula has been carried out in detail using multi-mode AFM system. Combining successful application of a novel AFM mode, i.e. Harmonix microscopy, the more detail elastic properties of spatula is brought to light. The results obtained show the variation of the mechanical properties on the hierarchical level of a seta, even for the different locations, pad and stalk of the spatula. A model, which has been validated using the existing experimental data and phenomena as well as theoretical predictions for geckos adhesion, crawling and self-cleaning of spatulae, is proposed in this paper. Through contrast of adhesive and craw ability of the gecko on the surfaces with different surface roughness, and measurement of the surface adhesive behaviors of Teflon, the most effective adhesion of the gecko is more dependent on the intrinsic properties of the surface which is adhered.


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