scholarly journals GENERAL NONCHEMOTACTIC MUTANTS OF CAULOBACTER CRESCENTUS

Genetics ◽  
1986 ◽  
Vol 114 (3) ◽  
pp. 717-730
Author(s):  
Bert Ely ◽  
Connie J Gerardot ◽  
Donna L Fleming ◽  
Suely L Gomes ◽  
Peter Frederikse ◽  
...  
Keyword(s):  
Genetic Analysis ◽  
Gene Cluster ◽  
Caulobacter Crescentus ◽  
Chemotactic Response ◽  
The Other ◽  
In Vitro Experiments ◽  
Methyltransferase Activity ◽  
Chemotaxis Proteins

ABSTRACT We have examined 35 mutants that have defects in general chemotaxis. Genetic analysis of these mutants resulted in the identification of at least eight che genes located at six different positions on the Caulobacter crescentus chromosome. The cheR, cheB and cheT genes appeared to be located in a three-gene cluster. Mutations in these three genes resulted in the inability of the flagellum to reverse the direction of rotation. Defects in the cheR gene resulted in a loss of the ability to methylate the methyl-accepting chemotaxis proteins. In vitro experiments showed that the lack of in vivo methylation in cheR mutants was due to the absence of methyltransferase activity. Defects in the cheB gene resulted in greatly reduced chemotaxis-associated methylation in vivo and a loss of methylesterase activity in vitro. The specific defects responsible for the lack of a chemotactic response have not been determined for the other identified che genes.

scholarly journals Induction of the Galactose Enzymes in Escherichia coli Is Independent of the C-1-Hydroxyl Optical Configuration of the Inducer d-Galactose

2008 ◽  
Vol 190 (24) ◽  
pp. 7932-7938 ◽  
Author(s):  
Sang Jun Lee ◽  
Dale E. A. Lewis ◽  
Sankar Adhya
Keyword(s):  
Escherichia Coli ◽  
Cell Wall ◽  
Carbon Source ◽  
The Other ◽  
Biosynthetic Pathways ◽  
In Vitro Experiments ◽  
Metabolizing Enzymes ◽  
Optical Configuration

ABSTRACT The two optical forms of aldohexose galactose differing at the C-1 position, α-d-galactose and β-d-galactose, are widespread in nature. The two anomers also occur in di- and polysaccharides, as well as in glycoconjugates. The anomeric form of d-galactose, when present in complex carbohydrates, e.g., cell wall, glycoproteins, and glycolipids, is specific. Their interconversion occurs as monomers and is effected by the enzyme mutarotase (aldose-1-epimerase). Mutarotase and other d-galactose-metabolizing enzymes are coded by genes that constitute an operon in Escherichia coli. The operon is repressed by the repressor GalR and induced by d-galactose. Since, depending on the carbon source during growth, the cell can make only one of the two anomers of d-galactose, the cell must also convert one anomer to the other for use in specific biosynthetic pathways. Thus, it is imperative that induction of the gal operon, specifically the mutarotase, be achievable by either anomer of d-galactose. Here we report in vivo and in vitro experiments showing that both α-d-galactose and β-d-galactose are capable of inducing transcription of the gal operon with equal efficiency and kinetics. Whereas all substitutions at the C-1 position in the α configuration inactivate the induction capacity of the sugar, the effect of substitutions in the β configuration varies depending upon the nature of the substitution; methyl and phenyl derivatives induce weakly, but the glucosyl derivative does not.

Influences of Metal Sprayed Coating Artificial Dental Root on the Vital Tissue

1998 ◽  
Author(s):  
M. Magome ◽  
K. Hara ◽  
K. Sano ◽  
Y. Ueda
Keyword(s):  
Plasma Spraying ◽  
Atmospheric Plasma Spraying ◽  
Aluminum Coating ◽  
The Other ◽  
Atmospheric Plasma ◽  
In Vitro Experiments ◽  
Wire Frame ◽  
Sprayed Coating

Abstract Artificial dental root material has been very important among all bio-materials. In this study, two different ways of aluminum coating are used. One is wire frame spraying, and the other is atmospheric plasma spraying. The affinity, stability, and connectivity of aluminum toxins with sprayed coating are used and the possibility for applying ways are examined. Acute toxic tests are performed clinically. Both in-vivo and in-vitro experiments are performed on a dog, and the results showed that aluminum sprayed coating is good as an artificial dental root material.

In vitro and in vivo effects of increased concentrations of free fatty acids on free thyroxin measurements as determined by five assays

1990 ◽  
Vol 36 (4) ◽  
pp. 611-613 ◽  
Author(s):  
R Sapin ◽  
J L Schlienger ◽  
F Grunenberger ◽  
F Gasser ◽  
J Chambron
Keyword(s):  
Fatty Acids ◽  
Free Fatty Acids ◽  
The Other ◽  
In Vitro Experiments ◽  
Fat Emulsion ◽  
Before And After ◽  
High Concentrations ◽  
In Vivo Effects

Abstract To compare in vitro and in vivo effects of increased concentrations of free fatty acids (FFA) on free thyroxin (FT4) values, we measured FT4 in three pooled sera supplemented with oleate and in serum from 18 euthyroid patients before and after an infusion of fat emulsion (Intralipid). We used five FT4 RIA kits: two two-step methods [Gammacoat, Baxter (GC); Ria-gnost, Behring (RG)], two analog RIAs [Amerlex-M, Amersham (AM); Coat-Ria, BioMérieux (CR)], and one kit with labeled antibodies [Amerlex-MAB*, Amersham (AA)]. In vitro, at the maximum oleate addition of 5 mmol/L, FT4 increased when measured by the GC and RG kits, decreased by the AM kit, and showed no significant change by the CR and AA kits. In vivo, post-Intralipid, FFA concentrations rose significantly and the FT4 changes agreed with the results of the in vitro experiments, except for the RG kit, for which FT4 increased in only nine patients. We conclude that in vitro oleate addition is useful to predict the in vivo effect of increased FFA on FT4 values; moreover, in serum from euthyroid subjects with high concentrations of FFA, FT4 analyzed with the CR or AA kits should better agree with normal results for thyrotropin than FT4 values measured with the other kits.

scholarly journals Nucleus-associated phosphorylation of Ins(1,4,5)P3 to InsP6 in Dictyostelium

1995 ◽  
Vol 312 (3) ◽  
pp. 911-917 ◽  
Author(s):  
J Van der Kaay ◽  
J Wesseling ◽  
P J M Van Haastert
Keyword(s):  
Hplc Analysis ◽  
The Other ◽  
In Vitro Experiments ◽  
Specific Sequence ◽  
Metabolic Route ◽  
And Function ◽  
Mutant Cells ◽  
Potential Substrate

Although many cells contain large amounts of InsP6, its metabolism and function is still largely unknown. In Dictyostelium lysates, the formation of InsP6 by sequential phosphorylation of inositol via Ins(3,4,6)P3 has been described [Stevens and Irvine (1990) Nature (London) 346, 580-583]; the second messenger Ins(1,4,5)P3 was excluded as a potential substrate or intermediate for InsP6 formation. However, we observed that mutant cells labelled in vivo with [3H]inositol showed altered labelling of both [3H]Ins(1,4,5)P3 and [3H]InsP6. In this report we demonstrate that Ins(1,4,5)P3 is converted into InsP6 in vitro by nucleus-associated enzymes, in addition to the previously described stepwise phosphorylation of inositol to InsP6 that occurs in the cytosol. HPLC analysis indicates that Ins(1,4,5)P3 is converted into InsP6 via sequential phosphorylation at the 3-, 6- and 2-positions. Ins[32P]P6, isolated from cells briefly labelled with [32P]Pi, was analysed using Paramecium phytase, which removes the phosphates of InsP6 in a specific sequence. The 6-position contained significantly more 32P radioactivity than the 4- or 5-positions, indicating that the 6-position is phosphorylated after the other two positions. The results from these in vivo and in vitro experiments demonstrate a metabolic route involving the phosphorylation of Ins(1,4,5)P3 via Ins(1,3,4,5)P4 and Ins(1,3,4,5,6)P5 to InsP6 in a nucleus-associated fraction of Dictyostelium cells.

scholarly journals The fermentation of soluble carbohydrates in rumen contents of cows given diets containing a large proportion of flaked maize

1969 ◽  
Vol 23 (3) ◽  
pp. 567-583 ◽  
Author(s):  
J. D. Sutton
Keyword(s):  
Acetic Acid ◽  
Propionic Acid ◽  
Butyric Acid ◽  
Volatile Fatty Acids ◽  
Valeric Acid ◽  
The Other ◽  
Soluble Carbohydrates ◽  
In Vitro Experiments

1. Studies were made of the fermentation of D-glucose, D-fructose, D-galactose, D-xylose, L-arabinose and sucrose by rumen contents from two cows fed 1 kg hay and 4 or 5 kg flaked maize once daily. The proportions of volatile fatty acids (VFA) in the rumen before addition of carbohydrates varied widely but on average acetic acid constituted about 52%, propionic acid about 29% and n-butyric acid about 13% of the total.2. In in vitro experiments, 896 mg of the monosaccharides and 851 mg sucrose were added to 150 g mixed rumen contents incubated for 2 h; the carbohydrates were added at 10 min intervals throughout the incubation on three occasions with each cow. Mean proportions of the carbohydrates fermented ranged from about 60% of the pentoses to about 85% of sucrose and glucose. Of the VFA produced from galactose and the pentoses, acetic acid constituted about 40%, propionic acid 45–55% and n-butyric acid 1–7%; very little n-valeric acid was produced. With the other carbohydrates results from the two cows differed, owing mainly to the production of appreciable amounts of n-valeric acid with one cow only. Acetic acid constituted about 40% of the VFA produced from fructose and sucrose, propionic acid 20–40%, n-butyric acid 14–22% and n-valeric acid up to 12%. The proportions of VFA produced from glucose were intermediate between the other two groups.3. Net recovery of carbon from fermented carbohydrate in VFA was about 35–45%. A further 1–6%, of fermented glucose, fructose and sucrose was recovered in lactic acid.4. In in vivo experiments, the monosaccharides only were infused into the rumen for 8 h at the rate of 200 g/h. Changes in the concentrations of substrates and products varied widely, owing to the variable basal fermentation, but changes in the proportions of VFA in the rumen were similar to those found in vitro.5. The results of the in vitro experiments were compared with those obtained in earlier experiments in which the same cows were given a diet containing 70% hay. Significant regressions (P < 0.05) were found between the molar proportions of acetic, propionic and n-valeric acids produced from the substrates and the proportions of these acids present in the rumen contents at the start of the incubations, but the relationships differed markedly among the different carbohydrates. Most of the significant regressions were positive but negative regressions for propionic acid production from fructose and sucrose with one cow suggested the existence of more complex interrelationships among two or more VFA.

in vitro experiments and in vivo implants to evaluate a new silicone-based polyurethane material for replacement of small vessels

2004 ◽  
Vol 14 (S3) ◽  
pp. 20-23 ◽  
Author(s):  
giorgio soldani ◽  
massimo bernabei ◽  
paola losi ◽  
adrian crucean ◽  
dante chiappino ◽  
...  
Keyword(s):  
Biomedical Applications ◽  
Chemical Properties ◽  
The Other ◽  
In Vitro Experiments ◽  
Small Vessels ◽  
Physical Chemical ◽  
Good Biocompatibility

the idea underscoring our proposed development is to take advantage of the good properties of both polyurethanes (pu) and silicones (pdms). the attributes which make polyurethanes attractive as materials for biomedical applications are their excellent physical–chemical properties, and their relatively good biocompatibility. against their use is the phenomenon of biodegradation that occurs after long-term implantation. silicones, on the other end, are known to have long-term biostability and good haemocompatibility subsequent to their use in several biomedical settings.

Enhancement of ADP-induced Platelet Aggregation by Adrenaline in Vivo and Its Prevention

1973 ◽  
Vol 29 (02) ◽  
pp. 490-498 ◽  
Author(s):  
Hiroh Yamazaki ◽  
Itsuro Kobayashi ◽  
Tadahiro Sano ◽  
Takio Shimamoto
Keyword(s):  
Platelet Count ◽  
Platelet Aggregation ◽  
Platelet Rich Plasma ◽  
The Other ◽  
Endothelial Surface ◽  
Important Interaction ◽  
Blood Coagulability ◽  
The Difference

SummaryThe authors previously reported a transient decrease in adhesive platelet count and an enhancement of blood coagulability after administration of a small amount of adrenaline (0.1-1 µg per Kg, i. v.) in man and rabbit. In such circumstances, the sensitivity of platelets to aggregation induced by ADP was studied by an optical density method. Five minutes after i. v. injection of 1 µg per Kg of adrenaline in 10 rabbits, intensity of platelet aggregation increased to 115.1 ± 4.9% (mean ± S. E.) by 10∼5 molar, 121.8 ± 7.8% by 3 × 10-6 molar and 129.4 ± 12.8% of the value before the injection by 10”6 molar ADP. The difference was statistically significant (P<0.01-0.05). The above change was not observed in each group of rabbits injected with saline, 1 µg per Kg of 1-noradrenaline or 0.1 and 10 µg per Kg of adrenaline. Also, it was prevented by oral administration of 10 mg per Kg of phenoxybenzamine or propranolol or aspirin or pyridinolcarbamate 3 hours before the challenge. On the other hand, the enhancement of ADP-induced platelet aggregation was not observed in vitro, when 10-5 or 3 × 10-6 molar and 129.4 ± 12.8% of the value before 10∼6 molar ADP was added to citrated platelet rich plasma (CPRP) of rabbit after incubation at 37°C for 30 second with 0.01, 0.1, 1, 10 or 100 µg per ml of adrenaline or noradrenaline. These results suggest an important interaction between endothelial surface and platelets in connection with the enhancement of ADP-induced platelet aggregation by adrenaline in vivo.

Applicability of Instability Index for In vitro Protein Stability Prediction

2019 ◽  
Vol 26 (5) ◽  
pp. 339-347 ◽  
Author(s):  
Dilani G. Gamage ◽  
Ajith Gunaratne ◽  
Gopal R. Periyannan ◽  
Timothy G. Russell
Keyword(s):  
Protein Stability ◽  
Caulobacter Crescentus ◽  
Effect Of Temperature ◽  
Instability Index ◽  
Dipeptide Composition ◽  
Experimental Conditions ◽  
The Stability ◽  
Vitro Protein

Background: The dipeptide composition-based Instability Index (II) is one of the protein primary structure-dependent methods available for in vivo protein stability predictions. As per this method, proteins with II value below 40 are stable proteins. Intracellular protein stability principles guided the original development of the II method. However, the use of the II method for in vitro protein stability predictions raises questions about the validity of applying the II method under experimental conditions that are different from the in vivo setting. Objective: The aim of this study is to experimentally test the validity of the use of II as an in vitro protein stability predictor. Methods: A representative protein CCM (CCM - Caulobacter crescentus metalloprotein) that rapidly degrades under in vitro conditions was used to probe the dipeptide sequence-dependent degradation properties of CCM by generating CCM mutants to represent stable and unstable II values. A comparative degradation analysis was carried out under in vitro conditions using wildtype CCM, CCM mutants and two other candidate proteins: metallo-β-lactamase L1 and α -S1- casein representing stable, borderline stable/unstable, and unstable proteins as per the II predictions. The effect of temperature and a protein stabilizing agent on CCM degradation was also tested. Results: Data support the dipeptide composition-dependent protein stability/instability in wt-CCM and mutants as predicted by the II method under in vitro conditions. However, the II failed to accurately represent the stability of other tested proteins. Data indicate the influence of protein environmental factors on the autoproteolysis of proteins. Conclusion: Broader application of the II method for the prediction of protein stability under in vitro conditions is questionable as the stability of the protein may be dependent not only on the intrinsic nature of the protein but also on the conditions of the protein milieu.

Oxytocin analogues with non-coded amino acid residues in position 8: [8-Neopentylglycine]oxytocin and [8-cycloleucine]oxytocin

1987 ◽  
Vol 52 (9) ◽  
pp. 2317-2325 ◽  
Author(s):  
Jan Hlaváček ◽  
Jan Pospíšek ◽  
Jiřina Slaninová ◽  
Walter Y. Chan ◽  
Victor J. Hruby
Keyword(s):  
Amino Acid ◽  
Solid Phase Synthesis ◽  
Solid Phase ◽  
The Other ◽  
Amino Acid Residues ◽  
Phase Synthesis ◽  
Other Hand ◽  
Fragment Condensation

[8-Neopentylglycine]oxytocin (II) and [8-cycloleucine]oxytocin (III) were prepared by a combination of solid-phase synthesis and fragment condensation. Both analogues exhibited decreased uterotonic potency in vitro, each being about 15-30% that of oxytocin. Analogue II also displayed similarly decreased uterotonic potency in vivo and galactogogic potency. On the other hand, analogue III exhibited almost the same potency as oxytocin in the uterotonic assay in vivo and in the galactogogic assay.

scholarly journals Development of a gel dedicated to gel immersion endoscopy

2021 ◽  
Vol 09 (06) ◽  
pp. E918-E924
Author(s):  
Tomonori Yano ◽  
Atsushi Ohata ◽  
Yuji Hiraki ◽  
Makoto Tanaka ◽  
Satoshi Shinozaki ◽  
...  
Keyword(s):  
Electrical Conductivity ◽  
Porcine Model ◽  
Ex Vivo ◽  
In Vitro Experiments ◽  
Efficacy And Safety ◽  
Coagulative Necrosis ◽  
Novel Technique ◽  
In Vivo Experiments

Abstract Backgrounds and study aims Gel immersion endoscopy is a novel technique to secure the visual field during endoscopy. The aim of this study was to develop a dedicated gel for this technique. Methods To identify appropriate viscoelasticity and electrical conductivity, various gels were examined. Based on these results, the dedicated gel “OPF-203” was developed. Efficacy and safety of OPF-203 were evaluated in a porcine model. Results  In vitro experiments showed that a viscosity of 230 to 1900 mPa·s, loss tangent (tanδ) ≤ 0.6, and hardness of 240 to 540 N/cm2 were suitable. Ex vivo experiments showed electrical conductivity ≤ 220 μS/cm is appropriate. In vivo experiments using gastrointestinal bleeding showed that OPF-203 provided clear visualization compared to water. After electrocoagulation of gastric mucosa in OPF-203, severe coagulative necrosis was not observed in the muscularis but limited to the mucosa. Conclusions OPF-203 is useful for gel immersion endoscopy.

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