scholarly journals BIOCHEMICAL STUDY OF CELLULAR ANTIGEN-ANTIBODY REACTION IN TISSUE CULTURE

1960 ◽  
Vol 112 (2) ◽  
pp. 237-247 ◽  
Author(s):  
Hideo Hayashi ◽  
Akira Tokuda ◽  
Keiji Udaka
Keyword(s):  
Culture Medium ◽  
Cultured Cells ◽  
Biochemical Study ◽  
Morphological Changes ◽  
Culture Fluid ◽  
Golgi Bodies ◽  
Optimum Ph ◽  
Protease Activation ◽  
Antibody Reaction ◽  
Antigen Antibody

The correlation between morphological and biochemical changes produced by the antigen-antibody reaction was studied in cultures of tissue monocytes taken from sensitized animals. The cells were grown under conditions which allowed collection of samples from the culture fluid as well as microscopic observation. Introduction of the antigen into the culture medium causes rapid release of a protease characterized by its susceptibility to sulfhydryl block and its optimum pH in the neutral range. Protease activation occurs simultaneously with morphological changes in the cytoplasm of the cultured cells. Delayed changes affecting the mitochondria and Golgi bodies appear after the peak of the proteolytic reaction and may be secondary to it. The gradual inactivation of the protease observed in the course of the antigen-antibody reaction will be discussed in a separate paper.

scholarly journals BIOCHEMICAL STUDY OF CELLULAR ANTIGEN-ANTIBODY REACTION IN TISSUE CULTURE

1960 ◽  
Vol 112 (2) ◽  
pp. 249-255 ◽  
Author(s):  
Akira Tokuda ◽  
Hideo Hayashi ◽  
Kinishiro Matsuba
Keyword(s):  
Tissue Culture ◽  
Protease Activity ◽  
Biochemical Study ◽  
Culture Fluid ◽  
Partial Purification ◽  
Trichloracetic Acid ◽  
Heat Stable ◽  
Antibody Reaction ◽  
Antigen Antibody ◽  
Cellular Antigen

Decrease in the protease activity of the culture fluid observed at later stages of the antigen-antibody reaction is believed to be due to the release of an inhibitor by the cells. The inhibitor was submitted to partial purification: it is heat-stable, non-precipitated by trichloracetic acid and non-dialyzable. It inhibits certain cellular and tissue proteases and papain but is inactive against trypsin. It is suggested that the balance between protease and anti-protease released may determine the intensity, extent, and duration of certain sensitization phenomena.

scholarly journals CONDITIONS INFLUENCING THE DISAPPEARANCE OF LIVING BACTERIA FROM THE BLOOD STREAM

1932 ◽  
Vol 55 (1) ◽  
pp. 121-137 ◽  
Author(s):  
Paul R. Cannon ◽  
F. L. Sullivan ◽  
E. F. Neckermann
Keyword(s):  
Intravenous Injection ◽  
Morphological Changes ◽  
Blood Stream ◽  
Active Immunization ◽  
Intracellular Digestion ◽  
Capillary Bed ◽  
Antibody Reaction ◽  
Antigen Antibody ◽  
Living Bacteria

1. The simultaneous intravenous injection into normal and actively immunized rabbits of equal quantities of living staphylococci or paratyphoid bacilli is followed by a distinctly accelerated rate of removal of the bacteria from the blood streams of the immune animals. 2. This altered reactivity is due essentially to specific active immunization. 3. The bacteria pass rapidly through the capillary bed of the lungs, extracellularly and dispersed for the most part, and become generalized through the blood stream. 4. The bacteria are quickly removed from the circulating blood in the immune animals and less rapidly in the normal ones, by various organs, particularly the liver and spleen, where they accumulate in enormous numbers, become adherent to the lining membrane of the sinusoids of the liver and apparently to the macrophages of the spleen and are phagocytosed by the macrophages and leucocytes in these organs. 5. Associated with this effect are morphological changes in the bacteria as shown by swelling, loss of staining power and evidences of increased cohesiveness and decreased viscosity, these changes being apparent as early as 2 minutes after their intravenous injection. 6. Inasmuch as these changes are not seen to a marked degree within the lungs or other organs, they are probably the result of a local antigen-antibody reaction of a bacteriotropic type in the two organs generally considered to be most actively concerned with the production of immune bodies. 7. By means of this accelerated bacteriotropic effect in the actively immunized animals, phagocytosis is facilitated and intracellular digestion of the bacteria is enhanced.

scholarly journals Fine Structure of Changes Produced in Cultured Cells Sampled at Specified Intervals During a Single Growth Cycle of Polio Virus

1958 ◽  
Vol 4 (3) ◽  
pp. 301-308 ◽  
Author(s):  
Frances Kallman ◽  
Robley C. Williams ◽  
Renato Dulbecco ◽  
Marguerite Vogt
Keyword(s):  
Fine Structure ◽  
Cultured Cells ◽  
Morphological Changes ◽  
Culture Fluid ◽  
Growth Cycle ◽  
Stage Ii ◽  
Virus Release ◽  
Polio Virus ◽  
Proper Size ◽  
Suspended Cultures

Primary suspended cultures of rhesus monkey kidney cells were infected with poliomyelitis virus, type 1 (Brunhilde strain). The release of virus from these cells over a one-step growth curve was correlated with their change in fine structure, as seen in the electron microscope. Most of the cells were infected nearly simultaneously, and morphological changes developed in the cells were sufficiently synchronous to be classified into three stages. The earliest change (stage I) became visible at a time when virus release into the culture fluid begins, some 3 hours after adsorption. Accentuation of the abnormal characteristics soon occurs, at 4 to 7 hours after adsorption, and results in stage II. Stage III represents the appearance of cells after their rate of virus release had passed its maximum, and probably the abnormal morphology of these cells reflects non-specific physiological damage. There seems to be consistency between the previously described cellular changes as seen under the light microscope and the finer scale changes reported here. Cytoplasmic bodies, called U bodies, were seen in large number at the time when the virus release was the most rapid (stage II). While these bodies are not of proper size to be considered polio virus, they seem to be specifically related to the infection. No evidence was found for the presence of particles that could even be presumptively identified with those of polio virus.

Correlation Between Cellular Functions and Morphological Changes of Human Trophoblast in Vitro

1975 ◽  
Vol 33 ◽  
pp. 600-601
Author(s):  
John C. Garancis ◽  
Robert O. Hussa ◽  
Michael T. Story ◽  
Donald Yorde ◽  
Roland A. Pattillo
Keyword(s):  
Electron Microscopy ◽  
Cellular Proliferation ◽  
Morphological Changes ◽  
Culture Fluid ◽  
Cellular Functions ◽  
Human Trophoblast ◽  
Transmission Electron ◽  
Marked Activity ◽  
Malignant Trophoblast

Human malignant trophoblast cells in continuous culture were incubated for 3 days in medium containing 1 mM N6-O2'-dibutyryl cyclic adenosine 3':5'-monophosphate (dibutyryl cyclic AMP) and 1 mM theophylline. The culture fluid was replenished daily. Stimulated cultures secreted many times more chorionic gonadotropin and estrogens than did control cultures in the absence of increased cellular proliferation. Scanning electron microscopy revealed remarkable surface changes of stimulated cells. Control cells (not stimulated) were smooth or provided with varying numbers of microvilli (Fig. 1). The latter, usually, were short and thin. The surface features of stimulated cells were considerably different. There was marked increase of microvilli which appeared elongated and thick. Many cells were covered with confluent polypoid projections (Fig. 2). Transmission electron microscopy demonstrated marked activity of cytoplasmic organelles. Mitochondria were increased in number and size; some giant forms with numerous cristae were observed.

Growth factors and the primary cilium in BALB/c 3T3 cells

1987 ◽  
Vol 45 ◽  
pp. 830-831
Author(s):  
R. W. Tucker ◽  
N. S. More ◽  
S. Jayaraman
Keyword(s):  
Growth Factors ◽  
Primary Cilium ◽  
Cultured Cells ◽  
Morphological Changes ◽  
Calcium Ionophore ◽  
Growth Stimulation ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
3T3 Cells ◽  
Microtubule Depolymerization

The mechanisms by which polypeptide growth factors Induce DNA synthesis in cultured cells is not understood, but morphological changes Induced by growth factors have been used as clues to Intracellular messengers responsible for growth stimulation. One such morphological change has been the transient disappearance of the primary cilium, a “9 + 0” cilium formed by the perinuclear centriole in interphase cells. Since calcium ionophore A23187 also produced both mitogenesis and ciliary changes, microtubule depolymerization might explain ciliary disappearance monitored by indirect immunofluorescence with anti-tubulin antibody. However, complete resorption and subsequent reformation of the primary cilium occurs at mitosis, and might also account for ciliary disappearance induced by growth factors. To settle this issue, we investigated the ultrastructure of the primary cilium using serial thin-section electron microscopy of quiescent BALB/c 3T3 cells before and after stimulation with serum.

scholarly journals A DNA Ligase from a Hyperthermophilic Archaeon with Unique Cofactor Specificity

2000 ◽  
Vol 182 (22) ◽  
pp. 6424-6433 ◽  
Author(s):  
Masaru Nakatani ◽  
Satoshi Ezaki ◽  
Haruyuki Atomi ◽  
Tadayuki Imanaka
Keyword(s):  
Biochemical Study ◽  
Protein Product ◽  
Optimum Concentration ◽  
Dna Ligase ◽  
Gene Encoding ◽  
Hyperthermophilic Archaeon ◽  
Dna Ligases ◽  
Cofactor Specificity ◽  
Thermophilic Archaeon ◽  
Optimum Ph

ABSTRACT A gene encoding DNA ligase (ligTk ) from a hyperthermophilic archaeon, Thermococcus kodakaraensisKOD1, has been cloned and sequenced, and its protein product has been characterized. ligTk consists of 1,686 bp, corresponding to a polypeptide of 562 amino acids with a predicted molecular mass of 64,079 Da. Sequence comparison with previously reported DNA ligases and the presence of conserved motifs suggested that Lig Tk was an ATP-dependent DNA ligase. Phylogenetic analysis indicated that Lig Tk was closely related to the ATP-dependent DNA ligase fromMethanobacterium thermoautotrophicum ΔH, a moderate thermophilic archaeon, along with putative DNA ligases fromEuryarchaeota and Crenarchaeota. We expressedligTk in Escherichia coli and purified the recombinant protein. Recombinant Lig Tk was monomeric, as is the case for other DNA ligases. The protein displayed DNA ligase activity in the presence of ATP and Mg2+. The optimum pH of Lig Tk was 8.0, the optimum concentration of Mg2+, which was indispensable for the enzyme activity, was 14 to 18 mM, and the optimum concentration of K+ was 10 to 30 mM. Lig Tk did not display single-stranded DNA ligase activity. At enzyme concentrations of 200 nM, we observed significant DNA ligase activity even at 100°C. Unexpectedly, Lig Tk displayed a relatively small, but significant, DNA ligase activity when NAD+ was added as the cofactor. Treatment of NAD+ with hexokinase did not affect this activity, excluding the possibility of contaminant ATP in the NAD+ solution. This unique cofactor specificity was also supported by the observation of adenylation of Lig Tk with NAD+. This is the first biochemical study of a DNA ligase from a hyperthermophilic archaeon.

scholarly journals Evaluation of Shear Horizontal Surface Acoustic Wave Biosensors Using “Layer Parameter” Obtained from Sensor Responses during Immunoreaction

Sensors ◽  
2021 ◽  
Vol 21 (14) ◽  
pp. 4924
Author(s):  
Koji Kano ◽  
Hiromi Yatsuda ◽  
Jun Kondoh
Keyword(s):  
Acoustic Wave ◽  
Surface Acoustic Wave ◽  
Horizontal Surface ◽  
Propagation Characteristics ◽  
Viscosity Change ◽  
Layer Parameter ◽  
Antibody Reaction ◽  
Antigen Antibody ◽  
Shear Horizontal ◽  
Molecular Dimensions

Shear horizontal surface acoustic wave (SH-SAW) biosensors measure the reaction of capture antibodies immobilized on the sensing surface to capture test molecules (antigens) by using the change in SH-SAW propagation characteristics. SH-SAW displacement exists not only on the SH-SAW propagating surface, but also partially penetrates the specimen liquid to a certain depth, which is determined by the liquid properties of the specimen and the operating frequency of the SH-SAW. This phenomenon is called viscosity penetration. In previous studies, the effect of viscosity penetration was not considered in the measurement of SH-SAW biosensors, and the mass or viscosity change caused by the specific binding of capture antibodies to the target antigen was mainly used for the measurement. However, by considering the effect of viscosity penetration, it was found that the antigen–antibody reaction could be measured and the detection characteristics of the biosensor could be improved. Therefore, this study aims to evaluate the detection properties of SH-SAW biosensors in the surface height direction by investigating the relationship between molecular dimensions and SH-SAW propagation characteristics, which are pseudo-changed by varying the diameter of gold nanoparticles. For the evaluation, we introduced a layer parameter defined by the ratio of the SH-SAW amplitude change to the SH-SAW velocity change caused by the antigen–antibody reaction. We found a correlation between the layer parameter and pseudo-varied molecular dimensions. The results suggest that SH-SAW does not only measure the mass and viscosity but can also measure the size of the molecule to be detected. This shows that SH-SAW biosensors can be used for advanced functionality.

Polygalacturonase isolated from the culture of the psychrophilic fungus Sclerotinia borealis

1997 ◽  
Vol 43 (5) ◽  
pp. 417-424 ◽  
Author(s):  
Toshihide Takasawa ◽  
Keiko Sagisaka ◽  
Koichi Yagi ◽  
Kyoko Uchiyama ◽  
Atsushi Aoki ◽  
...  
Keyword(s):  
Culture Medium ◽  
Enzyme Reaction ◽  
Wheat Seedling ◽  
Maximum Activity ◽  
Reducing Sugar ◽  
Pectic Acid ◽  
Snow Mold ◽  
Viscosity Change ◽  
Optimum Ph ◽  
Psychrophilic Fungus

A polygalacturonase was isolated from the culture medium of Sclerotinia borealis, a psychrophilic fungus that grows on lawn and wheat seedling under the snow in winter and induces the snow mold disease. Pectic acid was a better substrate of this enzyme than pectin when the activity was determined by measuring the reducing sugar produced. However, when the activity was measured by viscosity change, the viscosity of pectin decreased more rapidly than that of pectic acid. The results of viscosity change apparently indicate that the polygalacturonase catalyzes pectin hydrolysis as an endo-type enzyme. Highly methyl-esterified pectin was a poor substrate, as determined by measurements of reducing sugar production and viscosity change. It is suggested from the results that the methoxy group of pectin affects the polygalacturonase reaction. A reaction mechanism was proposed for the polygalacturonase reaction. Molecular mass of this enzyme was 40 kDa and its isoelectric point was pH 7.5. Optimum pH of the enzyme reaction was 4.5 and its optimum temperature was 40–50 °C. Thirty percent of the maximum activity was observed at 5 °C, but it was only slightly active above 60 °C. The activity was preserved for more than 2 years at 5 °C and pH 4.5, but it was lost when kept at room temperature overnight or heated at 50 °C for 30 min. The amino acid sequence of the N-terminal region of the psychrophilic polygalacturonase of Sclerotinia borealis is compared with those of polygalacturonases of mesophilic fungi. The function of this enzyme against the target plants is discussed with reference to the reaction of polygalacturonases of mesophilic fungi.Key words: polygalacturonase, pectin-hydrolyzing enzyme, psychrophilic fungi, snow mold disease, Sclerotinia borealis.

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