scholarly journals Stochasticity and positive feedback enable enzyme kinetics at the membrane to sense reaction size

2021 ◽  
Vol 118 (47) ◽  
pp. e2103626118
Author(s):  
Albert A. Lee ◽  
William Y. C. Huang ◽  
Scott D. Hansen ◽  
Neil H. Kim ◽  
Steven Alvarez ◽  
...  
Keyword(s):  
Positive Feedback ◽  
Membrane Surface ◽  
Strong Dependence ◽  
Reaction System ◽  
Stochastic Effects ◽  
Membrane Area ◽  
Copy Numbers ◽  
Lipid Kinases ◽  
The Mean ◽  
Activity State

Here, we present detailed kinetic analyses of a panel of soluble lipid kinases and phosphatases, as well as Ras activating proteins, acting on their respective membrane surface substrates. The results reveal that the mean catalytic rate of such interfacial enzymes can exhibit a strong dependence on the size of the reaction system—in this case membrane area. Experimental measurements and kinetic modeling reveal how stochastic effects stemming from low molecular copy numbers of the enzymes alter reaction kinetics based on mechanistic characteristics of the enzyme, such as positive feedback. For the competitive enzymatic cycles studied here, the final product—consisting of a specific lipid composition or Ras activity state—depends on the size of the reaction system. Furthermore, we demonstrate how these reaction size dependencies can be controlled by engineering feedback mechanisms into the enzymes.

Effect of Process Factors on Pervaporation Dehydration of Isopropanol

2017 ◽  
Vol 68 (6) ◽  
pp. 1302-1305
Author(s):  
Ali A. A. Al Janabi ◽  
Oana Cristina Parvulescu ◽  
Bogdan Trica ◽  
Tanase Dobre
Keyword(s):  
Ceramic Membrane ◽  
Membrane Surface ◽  
Water System ◽  
Surface Model ◽  
Feed Water ◽  
Mixture Flow ◽  
Water Separation ◽  
Membrane Area ◽  
Operation Temperature ◽  
Process Factors

The paper aimed at studying the performances of pervaporation separation of isopropanol-water system using a Pervatech ceramic membrane at various values of feed mixture flow rate (F=1000 kg/hr), feed water mass fraction (xF=0.1-0.2), operation temperature (t=60-90 �C), permeate pressure (pP=1000-9000 Pa) and water separation degree (sW=0.9, 0.95). Membrane total flux and separation factor were predicted applying a second order response surface model with 3 factors, i.e., xF, t and pP. An algorithm for estimating the membrane surface area was presented. Membrane area increased with sW and xF and its lowest values (A=13 m2 for xF=0.1 and A=24 m2 for xF=0.2) were attained for t=60 �C and pP=9000 Pa. These findings could be applied for optimizing the process of isopropanol dehydration by pervaporation.

Hydraulic consequences of enzymatic breakdown of grapevine pit membranes

2021 ◽  
Author(s):  
Ana Clara Fanton ◽  
Craig Brodersen
Keyword(s):  
Hydraulic Conductivity ◽  
Vascular System ◽  
Membrane Surface ◽  
Membrane Damage ◽  
Membrane Integrity ◽  
Biological Response ◽  
Plant Host ◽  
Important Species ◽  
Membrane Area ◽  
Pit Membrane

Abstract Xylella fastidiosa (Xf) is the xylem-dwelling bacterial agent associated with Pierce’s Disease (PD), which leads to significant declines in productivity in agriculturally important species like grapevine (Vitis vinifera). Xf spreads through the xylem network by digesting the pit membranes between adjacent vessels, thereby potentially changing the hydraulic properties of the stem. However, the effects of Xf on water transport varies depending on the plant host and the infection stage, presenting diverse outcomes. Here, we investigated the effects of polygalacturonase, an enzyme known to be secreted by Xf when it produces biofilm on the pit membrane surface, on stem hydraulic conductivity and pit membrane integrity. Experiments were performed on six grapevine genotypes with varying levels of PD resistance, with the expectation that pit membrane resistance to degradation by polygalacturonase may play a role in PD-resistance. Our objective was to study a single component of this pathosystem in isolation to better understand the mechanisms behind reported changes in hydraulics, thereby excluding the biological response of the plant to the presence of Xf in the vascular system. Pit membrane damage only occurred in stems perfused with polygalacturonase. Although the damaged pit membrane area was small (2-9% of the total pit aperture area), membrane digestion led to significant changes in the median air-seeding thresholds, and most importantly, shifted frequency distribution. Finally, enzyme perfusion also resulted in a universal reduction in stem hydraulic conductivity, suggesting the development of tyloses may not be the only contributing factor to reduced hydraulic conductivity in infected grapevine.

Desmosomes in hamster cheek pouch epithelium: their quantitative characterization during epithelial differentiation

1984 ◽  
Vol 66 (1) ◽  
pp. 411-429
Author(s):  
F.H. White ◽  
K. Gohari
Keyword(s):  
Plasma Membrane ◽  
Surface Area ◽  
Cheek Pouch ◽  
Unit Surface Area ◽  
Granular Cells ◽  
Hamster Cheek Pouch ◽  
Average Cell ◽  
Membrane Area ◽  
The Mean ◽  
Granular Layers

Desmosomes in stratified squamous epithelia appear to exhibit quantitative alterations during differentiation. In this work we use stereological and other morphometric methods to quantify these structures in epithelial cells from defined basal, spinous and granular strata. Hamster cheek pouch mucosa from five animals was processed for electron microscopy using strictly standardized techniques and a stratified random sampling procedure was used to obtain micrographs of cells from basal, spinous and granular layers. Stereological intersection counting techniques were used to determine for each layer the relative surface area of plasma membrane occupied by desmosomes (Ss), the number of desmosomes per unit surface area of plasma membrane (Ns), the mean individual desmosomal diameter (delta) and the mean individual desmosomal surface area (s). In addition, estimates of nuclear volume were obtained by direct measurement of nuclear profiles and volume-to-surface ratios were obtained by a combination of point and intersection counting, which enabled estimates for the volume (Vcell) and plasma membrane surface area (SPM) of the ‘average’ cell within each stratum to be acquired. Using this information, it was then possible to calculate both the total surface area (S) and the number (N) of desmosomes on the plasma membranes of average cells. The parameters Ss and Ns showed progressive increases between basal and granular layers, whereas values for delta and s were lower in granular cells when compared with basal and spinous cells. The parameters Vcell, SPM, S and N all increased progressively and significantly during differentiation. Between basal and granular layers, the mean cell volume and surface area had each increased approximately threefold, whereas the surface area and number of desmosomes on the average cell plasma membrane had increased approximately seven- and eleven-fold, respectively. Granular cells thus possess more numerous desmosomes, which occupy a greater proportion of the plasma membrane area but which are individually smaller, when compared with basal and spinous layers.

Orange juice ultrafiltration: characterisation of deposit layers and membrane surfaces after fouling and cleaning

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Nurul Hainiza Abd-Razak ◽  
Y. M. John Chew ◽  
Michael R. Bird
Keyword(s):  
Shear Stress ◽  
Orange Juice ◽  
Surface Coverage ◽  
Fluid Shear Stress ◽  
Membrane Surface ◽  
Regenerated Cellulose ◽  
Fluid Shear ◽  
Chemical Cleaning ◽  
Membrane Area ◽  
Software Analysis

Abstract The influence of feed condition and membrane cleaning during the ultrafiltration (UF) of orange juice for phytosterol separation was investigated. UF was performed using regenerated cellulose acetate (RCA) membranes at different molecular weight cut-off (MWCO) values with a 336 cm2 membrane area and a range of temperatures (10–40 °C) and different feed volumes (3–9 L). Fluid dynamic gauging (FDG) was applied to assess the fouling and cleaning behaviours of RCA membranes fouled by orange juice and cleaned using P3-Ultrasil 11 over two complete cycles. During the FDG testing, fouling layers were removed by fluid shear stress caused by suction flow. The cleanability was characterised by using ImageJ software analysis. A Liebermann-Buchard-based method was used to quantify the phytosterol content. The results show that RCA 10 kDa filters exhibited the best separation of phytosterols from protein in orange juice at 20 °C using 3 L feed with a selectivity factor of 17. Membranes that were fouled after two cycles showed higher surface coverage compared to one fouling cycle. The surface coverage decreased with increasing fluid shear stress from 0 to 3.9 Pa. FDG achieved 80–95% removal at 3.9 Pa for all RCA membranes. Chemical cleaning using P3-Ultrasil 11 altered both the membrane surface hydrophobicity and roughness. These results show that the fouling layer on RCA membranes can be removed by fluid shear stress without affecting the membrane surface modification caused by chemical cleaning.

scholarly journals A Systematic Study of Ammonia Recovery from Anaerobic Digestate Using Membrane-Based Separation

Membranes ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 19
Author(s):  
Fanny Rivera ◽  
Raúl Muñoz ◽  
Pedro Prádanos ◽  
Antonio Hernández ◽  
Laura Palacio
Keyword(s):  
Flow Rate ◽  
Membrane Surface ◽  
H2so4 Solution ◽  
Membrane Area ◽  
Recirculation Flow ◽  
Force Microscopy ◽  
Reservoir Volume ◽  
Atomic Force ◽  
Flat Sheet ◽  
Ammonia Recovery

Ammonia recovery from synthetic and real anaerobic digestates was accomplished using hydrophobic flat sheet membranes operated with H2SO4 solutions to convert ammonia into ammonium sulphate. The influence of the membrane material, flow rate (0.007, 0.015, 0.030 and 0.045 m3 h−1) and pH (7.6, 8.9, 10 and 11) of the digestate on ammonia recovery was investigated. The process was carried out with a flat sheet configuration at a temperature of 35 °C and with a 1 M, or 0.005 M, H2SO4 solution on the other side of the membrane. Polytetrafluoroethylene membranes with a nominal pore radius of 0.22 µm provided ammonia recoveries from synthetic and real digestates of 84.6% ± 1.0% and 71.6% ± 0.3%, respectively, for a membrane area of 8.6 × 10−4 m2 and a reservoir volume of 0.5 L, in 3.5 h with a 1 M H2SO4 solution and a recirculation flow on the feed side of the membrane of 0.030 m3 h−1. NH3 recovery followed first order kinetics and was faster at higher pHs of the H2SO4 solution and recirculation flow rate on the membrane feed side. Fouling resulted in changes in membrane surface morphology and pore size, which were confirmed by Atomic Force Microscopy and Air Displacement Porometry.

scholarly journals Coupled amidolytic assay for factor VII: its use with a clotting assay to determine the activity state of factor VII

Blood ◽  
1978 ◽  
Vol 52 (5) ◽  
pp. 978-988 ◽  
Author(s):  
U Seligsohn ◽  
B Osterud ◽  
SI Rapaport
Keyword(s):  
Healthy Subjects ◽  
Activity Ratio ◽  
Factor Xa ◽  
Factor Vii ◽  
Test Material ◽  
Chromogenic Substrate ◽  
Factor X ◽  
Transfusion Therapy ◽  
The Mean ◽  
Activity State

Abstract A coupled amidolytic assay for factor VII (VII) has been developed that when used with a clotting assay for VII enables detection of activated VII. In the assay, VII in a test material determines generation of factor Xa in a mixture of purified factor X, tissue factor, and calcium; factor Xa is measured with a chromogenic substrate. Factor VII activity in the coupled amidolytic assay (VIIam) correlated well with VII activity in a one-stage clotting assay (VIIc) in 57 healthy subjects, 5 patients with hereditary VII deficiency, and 11 patients with liver disease. Activation of plasma VII by kaolin, clotting, or cold strikingly increased VIIc but not VIIam levels. Thus the ratio VIIc/VIIam (VII activity ratio) is a measure of VII activation. In 27 warfarin-treated patients the mean VII activity ratio was significantly decreased, reflecting a greater decline in VIIc than in VIIam. This probably stems from partially carboxylated VII being able to act during the 3-min incubation of the amidolytic assay but unable to act rapidly enough to affect the clotting assay. Measurement of VIIc/VIIam should enable evaluation of the activity state of VII in thrombotic disorders and in components for transfusion therapy.

scholarly journals INVESTIGATION OF ENTEROCOCCUS FAECALIS POPULATION IN PATIENTS WITH POLYP AND COLORECTAL CANCER IN COMPARISON OF HEALTHY INDIVIDUALS

2019 ◽  
Vol 56 (2) ◽  
pp. 141-145 ◽  
Author(s):  
Maryam GERAVAND ◽  
Parviz FALLAH ◽  
Mojtaba Hedayat YAGHOOBI ◽  
Fatemeh SOLEIMANIFAR ◽  
Malihe FARID ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Enterococcus Faecalis ◽  
Copy Number ◽  
Healthy People ◽  
Control Group ◽  
Healthy Individuals ◽  
Faecal Flora ◽  
Copy Numbers ◽  
Significant Difference ◽  
The Mean

ABSTRACT BACKGROUND: Colorectal cancer is one of the most commonly diagnosed cancers around the world. One of the factors involved in the development of colorectal cancer is the changes in the normal flora of the intestine. OBJECTIVE: In this study, the mean copy number of Enterococcus faecalis in people with polyps and people with colorectal cancer has been evaluated in comparison with healthy controls. METHODS: In this study, 25 patients with colorectal cancer and 28 patients with intestinal polyps were selected and stool specimens were taken. In addition, 24 healthy individuals were selected as control group. Extraction of bacterial DNA from the stool sample were performed. The molecular methods of PCR for confirmation of standard strain and absolute Real Time PCR (qRT-PCR) method were used to evaluate the number of Enterococcus faecalis in the studied groups. RESULTS: The results of this study indicate that the mean copy number of Enterococcus faecalis in patients with colorectal cancer was 11.2x109 per gram of stool, and in patients with polyps was 9.4x108 per gram of stool. In healthy people, this number was 9x108 per gram of stool. There was a significant difference between the implicit copy numbers in the three groups. (P<0.05). CONCLUSION: Enterococcus faecalis in faecal flora of people with colorectal cancer was significantly higher than those with polyps and healthy people. This could potentially signify the ability of this bacterium to induce colorectal cancer. More studies are needed to prove this theory.

scholarly journals Modulated Response of Subtropical Gyres: Positive Feedback Loop, Subharmonic Modes, Resonant Solar and Orbital Forcing

2018 ◽  
Vol 6 (3) ◽  
pp. 107 ◽  
Author(s):  
Jean-Louis Pinault
Keyword(s):  
Positive Feedback ◽  
Feedback Loop ◽  
Rossby Waves ◽  
Resonance Phenomenon ◽  
Polar Coordinates ◽  
Western Boundary ◽  
Orbital Forcing ◽  
Positive Feedback Loop ◽  
The Mean

Evidence of long-term variability in the upper ocean has emerged for two decades. Most of the issues discussed raise a lot of questions. What is the driver of the decadal oscillation of rainfall in Europe that has been observed since the end of the 20th century? How to explain low-frequency variability as observed in the Atlantic Multidecadal Oscillation (AMO)? More generally, how does solar and orbital forcing occur during very long-term climate change? The observations suggest that both a positive feedback loop amplifies the effects of the insolation gradient on the climate system and a resonance phenomenon occurs, filtering out some frequencies in favour of others. Throughout this paper, some answers to these problems are given from a new concept based on the modulated response of subtropical gyres to solar and orbital forcing. Subtropical gyres turn out to be the main driver of long-term climate variability because they tightly control, via the western boundary currents, heat transport from the tropics into middle and high latitudes. Specifically, the theoretical foundations of long-period Rossby waves winding around the subtropical gyres are laid, suggested by the observations of persistent sea surface temperature anomalies at mid-latitudes. Multi-frequency Gyral Rossby Waves (GRWs) exhibit properties resulting from their annular structure and their coupling. Using a β-cone approximation, the momentum equations are solved in polar coordinates. The gradient β of the Coriolis parameter depends on the mean radius of the annulus and remains constant all around the latter. GRWs result from the variation in the Coriolis Effect with the mean radius of the annulus. The speed of the anti-cyclonically wind-driven circulation being higher than the phase velocity of cyclonically propagating GRWs, amplified forcing effects occur as well as resonances for periods consistent with the observations.

POSITIVE FEEDBACK ACTION OF OESTRADIOL ON GONADOTROPHIN RELEASE IN 15 DAY OLD FEMALE RATS

1977 ◽  
Vol 86 (2) ◽  
pp. 263-272 ◽  
Author(s):  
J. Kronibus ◽  
W. Wuttke
Keyword(s):  
Column Chromatography ◽  
Positive Feedback ◽  
Female Rats ◽  
Intact Control ◽  
Partial Separation ◽  
Ovx Rats ◽  
Serum Lh ◽  
The Mean ◽  
Intact Rats

ABSTRACT Female rats were ovariectomized (ovx), adrenalectomized (adx) or both (adx-ovx) on day 8 after birth. The serum gonadotrophin concentrations on day 15 were higher in ovx and adx-ovx rats than in sham-operated or untreated controls of the same age. Intact animals on day 15 had higher LH and FSH levels compared with adult, dioestrous levels, and a number of LH peaks were observed. After partial separation of oestradiol (LH 20 column chromatography) from other lipid substances which interfere with the radioimmunoassay for oestradiol, levels of oestradiol were undetectable in ovx and in adx-ovx animals on day 15 but concentrations were relatively high in intact or adx rats. To test whether the high gonadotrophin concentrations in 15-day-old intact rats were due to a positive feedback action of oestradiol, silastic tubes containing different amounts of oestradiol were implanted on day 8 at the time of adrenalectomy and ovariectomy. The mean serum LH and FSH concentrations were increased on day 15 in those animals in which silastic tube implantation resulted in physiological oestradiol levels. These elevated gonadotrophin values were due to a number of peak levels. Injection of 600 μg progesterone on day 15, 8 h before decapitation resulted in high FSH levels in all the implanted animals, whereas LH levels were still variable from one animal to another. This situation is very similar to that in intact control rats and it is concluded that the hypothalamo-pituitary axis in 15-day-old female rats reacts to an oestrogenic stimulus followed by a progestational reaction as does the adult "gonadostat". This would account for the premature, pre-ovulatory type of LH peaks.

Electronic structure of motoneurons in spinal cord slice cultures: a comparison of compartmental and equivalent cylinder models

1994 ◽  
Vol 72 (2) ◽  
pp. 861-871 ◽  
Author(s):  
D. Ulrich ◽  
R. Quadroni ◽  
H. R. Luscher
Keyword(s):  
Spinal Cord ◽  
Surface Area ◽  
Voltage Clamp ◽  
Time Constant ◽  
Membrane Surface ◽  
Input Resistance ◽  
Dendritic Trees ◽  
Membrane Surface Area ◽  
The Mean ◽  
Total Membrane

1. Voltage-clamp, current-clamp, and morphological data were obtained from visually identified motoneurons in organotypic cocultures of rat embryonic spinal cord, dorsal root ganglia, and skeletal muscle. The cells were injected with Biocytin during whole-cell patch-clamp recordings and stained with horseradish peroxidase. 2. The somata and dendritic trees of the cells were reconstructed with a semiautomatic reconstruction system. The motoneurons had a common multipolar shape. An elliptic soma gave rise to 3-9 stem dendrites with a mean diameter of 2.5 +/- 0.9 (SD) micron terminating in 24 +/- 7 dendritic endings. The mean total dendritic path length was 3,306 +/- 1,075 microns. The mean total membrane surface area was 15,594 +/- 10,404 microns 2 with a dendritic to somatic membrane surface area ratio of 3.4 +/- 1.4 (n = 7 cells). 3. The ratio between the sum of the diameters of the two daughter branches and the diameter of the parental branch each raised to the 3/2 power at all branch points was 1.3 +/- 0.28 (n = 8 cells). The dendritic trees of the cells tapered continuously from the soma to the distal ends. The mean normalized dendritic trunk parameter of all cells was 0.62 +/- 0.22. 4. The motoneurons had a mean input resistance RN of 498 +/- 374 M delta, a mean membrane time constant (tau m) of 22 +/- 4.6 ms, and a mean dendritic dominance (rho) of 2.7 +/- 0.86 (n = 5 cells). The mean electronic length (L) calculated from tau m and the slowest voltage-clamp time constant (tau VC1) was 0.7 +/- 0.04 (n = 7 cells). 5. The specific membrane capacitance (Cm) estimated from the charge of the capacitive current during a voltage step and the total membrane surface area was 1.08 +/- 0.3 microF/cm2 (n = 6 cells). 6. Compartmental computer models were constructed of individual cells. Experimental and simulated voltage transients were matched with Cm = 1 microF/cm2, a uniform membrane resistivity (Rm) = tau m/Cm and a cytosolic resistivity (Ri) of 308 +/- 39 omega.cm (n = 3 cells). 7. The mean electrotonic length of the dendritic paths was 0.83 +/- 0.2 (n = 5 cells). The mean input resistance at the dendritic terminals (RT) was 1,413 +/- 260 M omega. Synaptic conductances were applied at all distal dendritic compartments of the model cells. The resulting synaptic currents were calculated at the input site and at the soma. The mean transient current attenuation ratio was 4.7 +/- 1.7 under idealized voltage-clamp conditions.(ABSTRACT TRUNCATED AT 400 WORDS)

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