Intercellular Invertase in Developing Peach Mesocarp

1988 ◽  
Vol 15 (3) ◽  
pp. 377 ◽  
Author(s):  
TD Ugalde ◽  
DJ Chalmers ◽  
PH Jerie

Acid invertase (β-fructofuranosidase, EC 3.2.1.26) was extracted from peach mesocarp (Prunus persica (L.) Batsch) using a range of extraction conditions. The enzyme always was attached to insoluble particles in the crude homogenate and was bound by a mechanism that could not have arisen during extraction. The activity in the insoluble fraction made up (essentially) all of the total activity extracted from the tissue and was the same as the activity shown by whole tissue slices placed directly into the assay solution. These results demonstrate that most of the acid invertase in developing peach mesocarp is located outside the cell. The amount of this enzyme, as measured in vitro, did not change during development at times when the rate of dry matter increase was changing rapidly. Either the action of intercellular invertase is not associated with the control of dry matter accumulation in peach mesocarp, or control is effected through activity of the enzyme in vivo, not its synthesis or degradation.

2021 ◽  
Vol 22 (5) ◽  
pp. 2285
Author(s):  
Thu Hang Lai ◽  
Susann Schröder ◽  
Magali Toussaint ◽  
Sladjana Dukić-Stefanović ◽  
Mathias Kranz ◽  
...  

The adenosine A2A receptor (A2AR) represents a potential therapeutic target for neurodegenerative diseases. Aiming at the development of a positron emission tomography (PET) radiotracer to monitor changes of receptor density and/or occupancy during the A2AR-tailored therapy, we designed a library of fluorinated analogs based on a recently published lead compound (PPY). Among those, the highly affine 4-fluorobenzyl derivate (PPY1; Ki(hA2AR) = 5.3 nM) and the 2-fluorobenzyl derivate (PPY2; Ki(hA2AR) = 2.1 nM) were chosen for 18F-labeling via an alcohol-enhanced copper-mediated procedure starting from the corresponding boronic acid pinacol ester precursors. Investigations of the metabolic stability of [18F]PPY1 and [18F]PPY2 in CD-1 mice by radio-HPLC analysis revealed parent fractions of more than 76% of total activity in the brain. Specific binding of [18F]PPY2 on mice brain slices was demonstrated by in vitro autoradiography. In vivo PET/magnetic resonance imaging (MRI) studies in CD-1 mice revealed a reasonable high initial brain uptake for both radiotracers, followed by a fast clearance.


1977 ◽  
Vol 57 (2) ◽  
pp. 279-288 ◽  
Author(s):  
S. O. THORLACIUS

Digestibility and intake of diets containing 8, 28, 48 or 68% ground wheat straw plus ground crested wheatgrass and rapeseed meal, and diets containing 33, 48, 63 and 78% ground crested wheatgrass plus barley and rapeseed meal was measured with four yearling wethers per diet. Digestible energy (DE) content ranged from 2.07 to 2.95 kcal/g dry matter (DM) and dry matter digestibility (DMD) (%) from 48.7 to 71.1%. Regression of DE intake y (kcal/w0.75kg/d) on DE content (x) was curvilinear; y = −2,133 + 1,626x − 277.9x2, r = 0.996, P < 0.01, SE = ± 7.3. There was also a curvilinear relationship between diet density, as fed, (x) g (DM)/ml and DMD (%), y = 9.057 + 364.1x − 530.0x2, r = 0.970, P < 0.01, SE = ± 2.4. A linear regression equation was calculated over the DE range (2.07–2.52) for which there was an obvious increase in DE intake with increasing diet DE content; y = −700.6 + 361x, r = 0.994, P < 0.01, SE = ± 9.4, y = DE intake (kcal/w0.75kg/d), x = DE [kcal/g (DM)]. Using this regression equation and assuming a linear increase in DE intake with increase in diet DE content up to a point at which the apparent energy demand of the animal is satisfied gave a more accurate prediction of DE intake than when the curvilinear regression equation, y = −2,133 + 1,626x − 277.9x2, was used empirically. Accuracy of the prediction was further improved by expressing DE/unit ration volume instead of per unit DM. The sheep used in the present experiments had an apparent energy demand of 230 kcal/w0.75kg/day which was met at diet DE contents above 0.48 kcal/ml or 2.6 kcal/g (DM). There was a high correlation between in vivo DE content of the diet, y [kcal/g (DM)] and in vitro (x) digestible organic matter content, x, (%), r = 0.991, P < 0.01, y = 0.38 + 0.037x, SE = ± 0.04.


Author(s):  
Xiaoming He ◽  
Shawn Mcgee ◽  
James E. Coad ◽  
Paul A. Iaizzo ◽  
David J. Swanlund ◽  
...  

In this paper, we report on the characterization of microwave therapy of normal porcine kidneys both in vitro and in vivo. This technology is being developed for eventual use in the treatment of small renal cell carcinoma (RCC) by minimally invasive procedures. During experiments, microwave energy was applied through an interstitial microwave probe (Urologix, Plymouth, MN) to the kidney cortex with occasional involvement of the kidney medulla. The thermal histories at several locations were recorded. After treatment, the kidneys were bisected and small tissue slices were cut out at approximately the same depth as the thermal probes. The tissue slices were further processed for histological study. Both cellular injury and the area of microvascular stasis were quantitatively evaluated by histology. Absolute rate kinetic models of cellular injury and vascular stasis were developed and fit to this data. A 3-D finite element thermal model based on the Pennes Bioheat equation was developed and solved using a commercial software package (ANSYS, V5.7). The Specific Absorption Rate (SAR) of the microwave probe was measured experimentally in tissue equivalent gel-like solution. The thermal model was first validated by the measured in vitro thermal histories. It was then used to determine the blood perfusion term in vivo.


1997 ◽  
Vol 272 (3) ◽  
pp. L479-L485 ◽  
Author(s):  
M. Ikegami ◽  
T. R. Korfhagen ◽  
M. D. Bruno ◽  
J. A. Whitsett ◽  
A. H. Jobe

In the present study we asked if surfactant metabolism was altered in surfactant protein (SP) A-deficient mice in vivo. Although previous studies in vitro demonstrated that SP-A modulates surfactant secretion and reuptake by type II cells, mice made SP-A deficient by homologous recombination grow and reproduce normally and have normal lung function. Alveolar and lung tissue saturated phophatidylcholine (Sat PC) pools were 50 and 26% larger, respectively, in SP-A(-/-) mice than in SP-A(+/+) mice. Radiolabeled choline and palmitate incorporation into lung Sat PC was similar both in vivo and for lung tissue slices in vitro from SP-A(+/+) and SP-A(-/-) mice. Percent secretion of radiolabeled Sat PC was unchanged from 3 to 15 h, although SP-A(-/-) mice retained more labeled Sat PC in the alveolar lavages at 48 h (consistent with the increased surfactant pool sizes). Clearance of radiolabeled dipalmitoylphosphatidylcholine and SP-B from the air spaces after intratracheal injection was similar in SP-A(-/-) and SP-A(+/+) mice. Lack of SP-A had minimal effects on the overall metabolism of Sat PC or SP-B in mice.


1975 ◽  
Vol 23 (1) ◽  
pp. 3-9
Author(s):  
S. Tamminga ◽  
C.J. van der Koelen

1. Grass from the same sward was ensiled without additive, with 14.6 g formic acid/100 g crude protein or 10.8 g formic acid and 10.6 g formaldehyde/100 g crude protein. Similar grass was dried and pelleted. Drying or ensiling with the mixture reduced solubility of N in the preserved grass but formic acid increased it, and ensiling without additive increased it even more. Apparent digestibility of N in the rumen of cows tended to decrease with decrease in solubility. Digestibility in vitro of the mixed diet given to the cows, calculated from digestibility of the separate components, agreed well with the values in vivo for diets with silages, but was high for that with dried grass. (Abstract retrieved from CAB Abstracts by CABI’s permission)


1972 ◽  
Vol 23 (1) ◽  
pp. 25 ◽  
Author(s):  
DC Brown ◽  
JC Radcliffe

Twenty experimental silages were made from seven pasture species at different stages of maturity. In vivo dry matter, organic matter, and energy ad libitum intakes and digestibilities of the silages were determined with standardized pairs of Merino wethers. The following chemical characteristics of the silages were measured: nitrogen, ammonia nitrogen, total titratable acids, acetic, propionic, butyric, and lactic acids, total volatiles lost during oven drying, lactic acid as a percentage of the total organic acids, pH, acid pepsin dry matter disappearance, dry matter content, and in vitro digestibility and rate of digestion. When all 20 silages were considered, energy intakes on a body weight basis were significantly related to silage pH (r = 0.55) and rate of in vitro digestion (r = 0.58). When the five legume silages were removed from the analysis and only the 15 grass-dominant silages were considered, dry matter intakes were significantly related to acetic (r = –0.57) and propionic acid (r = –0.55) concentrations. Multiple regression analyses did not significantly increase the accuracy of predicting intake. The results suggested that silage intake was negatively related to the degree of fermentation that occurred during the ensiling process.


1996 ◽  
Vol 135 (1) ◽  
pp. 37-51 ◽  
Author(s):  
M Hirao ◽  
N Sato ◽  
T Kondo ◽  
S Yonemura ◽  
M Monden ◽  
...  

The ERM proteins, ezrin, radixin, and moesin, are involved in the actin filament/plasma membrane interaction as cross-linkers. CD44 has been identified as one of the major membrane binding partners for ERM proteins. To examine the CD44/ERM protein interaction in vitro, we produced mouse ezrin, radixin, moesin, and the glutathione-S-transferase (GST)/CD44 cytoplasmic domain fusion protein (GST-CD44cyt) by means of recombinant baculovirus infection, and constructed an in vitro assay for the binding between ERM proteins and the cytoplasmic domain of CD44. In this system, ERM proteins bound to GST-CD44cyt with high affinity (Kd of moesin was 9.3 +/- 1.6nM) at a low ionic strength, but with low affinity at a physiological ionic strength. However, in the presence of phosphoinositides (phosphatidylinositol [PI], phosphatidylinositol 4-monophosphate [4-PIP], and phosphatidylinositol 4.5-bisphosphate [4,5-PIP2]), ERM proteins bound with a relatively high affinity to GST-CD44cyt even at a physiological ionic strength: 4,5-PIP2 showed a marked effect (Kd of moesin in the presence of 4,5-PIP2 was 9.3 +/- 4.8 nM). Next, to examine the regulation mechanism of CD44/ERM interaction in vivo, we reexamined the immunoprecipitated CD44/ERM complex from BHK cells and found that it contains Rho-GDP dissociation inhibitor (GDI), a regulator of Rho GTPase. We then evaluated the involvement of Rho in the regulation of the CD44/ERM complex formation. When recombinant ERM proteins were added and incubated with lysates of cultured BHK cells followed by centrifugation, a portion of the recombinant ERM proteins was recovered in the insoluble fraction. This binding was enhanced by GTP gamma S and markedly suppressed by C3 toxin, a specific inhibitor of Rho, indicating that the GTP form of Rho in the lysate is required for this binding. A mAb specific for the cytoplasmic domain of CD44 also markedly suppressed this binding, identifying most of the binding partners for exogenous ERM proteins in the insoluble fraction as CD44. Consistent with this binding analysis, in living BHK cells treated with C3 toxin, most insoluble ERM proteins moved to soluble compartments in the cytoplasm, leaving CD44 free from ERM. These findings indicate that Rho regulates the CD44/ERM complex formation in vivo and that the phosphatidylinositol turnover may be involved in this regulation mechanism.


1997 ◽  
Vol 1997 ◽  
pp. 197-197
Author(s):  
R. Sanderson ◽  
S.J. Lister ◽  
A. Sargeant ◽  
M.S. Dhanoa

The objectives of this study were a) to examine the effect of particle size and silage dry matter (DM) content on the rate and pattern of fermentation of fresh silages in vitro as an aid to modelling the in vivo situation and b) to compare the rate and pattern of fermentation of fresh silage samples with those obtained for freeze-dried material.


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