Two-dimensional electrophoretic analysis of in vivo and in vitro synthesis of proteins in peas inoculated with compatible and incompatible Fusarium solani

1982 ◽  
Vol 20 (1) ◽  
pp. 99-107 ◽  
Author(s):  
Wendy Wagoner ◽  
David C. Loschke ◽  
Lee A. Hadwiger
Author(s):  
U. Aebi ◽  
L.E. Buhle ◽  
W.E. Fowler

Many important supramolecular structures such as filaments, microtubules, virus capsids and certain membrane proteins and bacterial cell walls exist as ordered polymers or two-dimensional crystalline arrays in vivo. In several instances it has been possible to induce soluble proteins to form ordered polymers or two-dimensional crystalline arrays in vitro. In both cases a combination of electron microscopy of negatively stained specimens with analog or digital image processing techniques has proven extremely useful for elucidating the molecular and supramolecular organization of the constituent proteins. However from the reconstructed stain exclusion patterns it is often difficult to identify distinct stain excluding regions with specific protein subunits. To this end it has been demonstrated that in some cases this ambiguity can be resolved by a combination of stoichiometric labeling of the ordered structures with subunit-specific antibody fragments (e.g. Fab) and image processing of the electron micrographs recorded from labeled and unlabeled structures.


Author(s):  
Leydi Miguel-Ferrer ◽  
Omar Romero-Arenas ◽  
Petra Andrade-Hoyos ◽  
Primo Sánchez-Morales ◽  
José Antonio Rivera-Tapia ◽  
...  

El chile es la segunda hortaliza de mayor producción en México. El objetivo de la investigación fue evaluar la actividad antagónica <em>in vitro</em> e <em>in vivo</em> de <em>Trichoderma harzianum</em> (T-H4) y <em>T. koningiopsis</em> (T-K11) <em>versus Fusarium solani</em> (MX-MIC 798) en la germinación y establecimiento de plántula de chile Miahuateco. Se utilizó la técnica de cultivo dual para determinar el porcentaje de inhibición de crecimiento radial (PICR) de la cepa MX-MIC 798. Además, se analizó el porcentaje de germinación en semillas de chile Miahuateco en vivero, así como la mortalidad de plántulas y grado de severidad a los 40 días después del trasplante (ddt) en Santa María Tecomavaca, Oaxaca, a través de biocontrol y control químico (Mancozeb 80®). La cepa T-H4 presentó el nivel antagónico PICR más alto (53.3%) <em>in vitro</em> y clase II en la escala de Bell, asimismo obtuvo 82% de germinación en semillas de chile Miahuateco en vivero y 48% de mortalidad en campo; de manera que igualó al control químico y superó a <em>T. koningiopsis</em> T-K11. La actividad antifúngica de <em>Trichoderma</em> spp., ofrecen una alternativa para el biocontrol de la marchitez y necrosis en raíz del cultivo de chile Miahuateco causada por <em>F. solani</em> MX-MIC 798.


2021 ◽  
Author(s):  
Rebecca L Pinals ◽  
Nicholas Ouassil ◽  
Jackson Travis Del Bonis-O'Donnell ◽  
Jeffrey W Wang ◽  
Markita P Landry

Engineered nanoparticles are advantageous for numerous biotechnology applications, including biomolecular sensing and delivery. However, testing the compatibility and function of nanotechnologies in biological systems requires a heuristic approach, where unpredictable biofouling often prevents effective implementation. Such biofouling is the result of spontaneous protein adsorption to the nanoparticle surface, forming the "protein corona" and altering the physicochemical properties, and thus intended function, of the nanotechnology. To better apply engineered nanoparticles in biological systems, herein, we develop a random forest classifier (RFC) trained with proteomic mass spectrometry data that identifies which proteins adsorb to nanoparticles. We model proteins that populate the corona of a single-walled carbon nanotube (SWCNT)-based optical nanosensor. We optimize the classifier and characterize the classifier performance against other models. To evaluate the predictive power of our model, we then apply the classifier to rapidly identify and experimentally validate proteins with high binding affinity to SWCNTs. Using protein properties based solely on amino acid sequence, we further determine protein features associated with increased likelihood of SWCNT binding: proteins with high content of solvent-exposed glycine residues and non-secondary structure-associated amino acids. Furthermore, proteins with high leucine residue content and beta-sheet-associated amino acids are less likely to form the SWCNT protein corona. The classifier presented herein provides an important tool to undertake the otherwise intractable problem of predicting protein-nanoparticle interactions, which is needed for more rapid and effective translation of nanobiotechnologies from in vitro synthesis to in vivo use.


1980 ◽  
Vol 45 (2) ◽  
pp. 435 ◽  
Author(s):  
Milutin Drobac ◽  
Harry Rakowski ◽  
Brian W. Gilbert ◽  
Michael X. Glynn ◽  
Malcolm D. Silver

1964 ◽  
Vol 22 (1) ◽  
pp. 227-258 ◽  
Author(s):  
Burton Goldberg ◽  
Howard Green

In vitro synthesis of collagen by established mouse fibroblast lines has been examined by electron microscopy. During rapid growth (log phase), when collagen could not be detected in the cultures, the cells lacked a well developed granular ergastoplasm and Golgi system. Upon cessation of growth (stationary phase), collagen accumulated in the cultures and the cells demonstrated highly developed granular and smooth ergastoplasm. Collagen appeared to be synthesized in the rough-surfaced endoplasmic reticulum and to be transported as a soluble protein to the cell surface by vesicular elements of the agranular ergastoplasm. Fusion of the limiting membranes of these vesicles with the cell membrane permitted the discharge of the soluble collagen into the extracellular space, where fibrils of two diameter distributions formed. The secretion of collagen is concluded to be of the merocrine type. Alternative theories of collagen secretion are discussed and the data for established lines compared with the results of other in vitro and in vivo studies of collagen fibrillogenesis.


2012 ◽  
Vol 9 (77) ◽  
pp. 3288-3302 ◽  
Author(s):  
Neha Arya ◽  
Viren Sardana ◽  
Meera Saxena ◽  
Annapoorni Rangarajan ◽  
Dhirendra S. Katti

Owing to the reduced co-relationship between conventional flat Petri dish culture (two-dimensional) and the tumour microenvironment, there has been a shift towards three-dimensional culture systems that show an improved analogy to the same. In this work, an extracellular matrix (ECM)-mimicking three-dimensional scaffold based on chitosan and gelatin was fabricated and explored for its potential as a tumour model for lung cancer. It was demonstrated that the chitosan–gelatin (CG) scaffolds supported the formation of tumoroids that were similar to tumours grown in vivo for factors involved in tumour-cell–ECM interaction, invasion and metastasis, and response to anti-cancer drugs. On the other hand, the two-dimensional Petri dish surfaces did not demonstrate gene-expression profiles similar to tumours grown in vivo . Further, the three-dimensional CG scaffolds supported the formation of tumoroids, using other types of cancer cells such as breast, cervix and bone, indicating a possible wider potential for in vitro tumoroid generation. Overall, the results demonstrated that CG scaffolds can be an improved in vitro tool to study cancer progression and drug screening for solid tumours.


1990 ◽  
Vol 259 (5) ◽  
pp. E723-E728
Author(s):  
J. A. Durr ◽  
W. H. Hoffman ◽  
J. Hensen ◽  
A. H. Sklar ◽  
T. el Gammal ◽  
...  

Osmoregulation of arginine vasopressin (AVP) is altered in diabetic ketoacidosis (DKA). With hyperglycemia, the AVP-plasma sodium (PNa) curve is displaced to the left, whereas the AVP-osmolality (Posm) curve is displaced to the right. The shift in the Na curve is explained by either resetting of the Na set point or by glucose acting as a nonpermeable solute, substituting for Na. Conversely, putative unmeasured solutes that, like urea, fail to affect AVP have been postulated to account for the right shift in the AVP-Posm curve. Therefore the respective roles of Posm = sigma [Xi] and plasma tonicity (Pton = sigma [sigmaiXi]), i.e., the sum of concentrations of all solutes [Xi] corrected (Pton) or not (Posm) for their relative cell permeability (sigma i), were studied in DKA. Indeed, Posm = sigma [Xi] exceeds Pton = sigma [sigma iXi] in DKA, since sigma i less than 1 for glucose. Potential determinants of AVP release (Posm, Pton, and PNa) were monitored in 7 patients with DKA. Conventional correlation analysis and two-dimensional (2D) graphs reproduced the paradox of an opposite shift in PNa and Posm set points for AVP release. However, by using the concept of tonicity instead of osmolality, 3D plots instead of 2D graphs, and multiple regressions instead of correlations, the AVP-PNa and AVP-Pton curves did not appear displaced. The concept of tonicity resolved the paradox of both osmolality and Na thresholds reset in opposite directions. Indeed, in states where a solute like glucose (with sigma less than 1) contributes substantially to plasma osmolality, Posm measured in vitro by the osmometer greatly exceeds Pton perceived in vivo by the osmoreceptor.


1983 ◽  
Vol 61 (10) ◽  
pp. 1079-1084 ◽  
Author(s):  
Fred W. Keeley ◽  
Dorothy J. Johnson

The rate of in vitro synthesis of soluble and insoluble elastin by thoracic aorta of 2-day-old chicks has been measured in absolute terms. In the absence of β-aminopropionitrile (βAPN), the steady state level of soluble elastin was 120 pmol/100 mg of aortic tissue or 3.7 μg/whole aorta segment. The rate of synthesis of elastin in vitro was approximately 130 μg/day per whole aorta segment. This is three- to four-fold lower than the estimated rate of in vivo synthesis for a comparable segment of aortic tissue at the same stage of development. Pulse-chase experiments suggested that this difference was not due to in vitro proteolysis of a significant proportion of the newly synthesized soluble elastin, but rather that the conversion of soluble to insoluble elastin was both rapid and efficient. These experiments also indicated the presence in aortic tissue of a substantial pool of elastin of intermediate solubility. Although inclusion of βAPN in the incubation medium resulted in an increase in the amount of soluble elastin in aortic tissue, the rate of accumulation of newly synthesized soluble elastin in the presence of this inhibitor of cross-linking was not linear, but decreased with incubation time. Furthermore, although βAPN effectively suppressed the appearance of insoluble elastin for at least 2 h, some escape from the effect of this inhibitor was seen with further incubation. In general, βAPN significantly depressed elastin synthesis.


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