scholarly journals MOLECULAR WEIGHT, ELECTROCHEMICAL AND BIOLOGICAL PROPERTIES OF TUBERCULIN PROTEIN AND POLYSACCHARIDE MOLECULES

1938 ◽  
Vol 68 (3) ◽  
pp. 413-438 ◽  
Author(s):  
Florence B. Seibert ◽  
Kai O. Pedersen ◽  
Arne Tiselius
Keyword(s):  
Molecular Weight ◽  
Nucleic Acid ◽  
Guinea Pigs ◽  
Culture Media ◽  
Spherical Shape ◽  
Biological Properties ◽  
Natural State ◽  
Local Skin ◽  
Precipitin Reaction ◽  
Bovine Strain

Studies have been made by means of sedimentation in the ultracentrifuge, and by diffusion and electrophoresis, to determine the molecular weights and homogeneity of the tuberculin protein and polysaccharide molecules as found in their natural state in the unchanged filtrates from culture media after growth of tubercle bacilli. These results have been compared with data obtained on fractions isolated by chemical procedures from them or from old tuberculin. By means of electrophoresis in the Tiselius apparatus it was possible to separate the protein from the polysaccharide, as these two fractions occur naturally in the original culture medium filtrates of acid-fast bacilli. The protein from the bovine strain of bacillus proved to be homogeneous in sedimentation (S20 = 1.6), diffusion (D20 = 12.0) and electrophoresis, with a molecular weight of about 10,000. The tuberculin polysaccharide isolated in electrophoresis appeared to be practically the same in sedimentation and in precipitin reaction as the polysaccharide isolated by chemical procedure. The latter proved to be homogeneous in sedimentation (S20 = 1.6) and diffusion (D20 = 11.0) with a molecular weight of about 9000. A practically homogeneous protein was isolated from the culture filtrate of the human tubercle bacillus H 37 by fractional ammonium sulfate precipitation, with a molecular weight of 32,000 (S20 = 3.3; D20 = 8.2). It was electrochemically homogeneous, with an isoelectric point at pH 4.3 and an isoionic point at pH 4.7. It could be dried or frozen with no loss in homogeneity. It was highly antigenic in the precipitin reaction and produced the anaphylactic type of local skin reaction in tuberculous guinea pigs, in contrast to the true tuberculin type of reaction caused by a purified PPD fraction. Furthermore death resulted in tuberculous guinea pigs from intracutaneous injection of exceptionally small amounts. A protein with molecular weight of about 17,000 was isolated from the filtrate from cultures of the timothy bacillus. The nucleic acid originally occurring in old tuberculin (OT) seems to be responsible for the high electrochemical mobility observed. From OT and the PPD made from it, potent but non-antigenic molecules of 16,000 and 9000 weight and with a low content of nucleic acid were isolated. With increase in size these deviated much from the normal compact spherical shape, and aggregation was evident from the tendency toward gel formation. The smallest molecule (9000) was homogeneous (S20 = 1.0; D20 = 10.0) and had lost some tuberculin potency.

scholarly journals ETIOLOGY OF YELLOW FEVER

1919 ◽  
Vol 30 (1) ◽  
pp. 13-29 ◽  
Author(s):  
Hideyo Noguchi
Keyword(s):  
Yellow Fever ◽  
Pure Culture ◽  
Guinea Pigs ◽  
Culture Media ◽  
Morphological Characteristics ◽  
Biological Properties ◽  
Freezing And Thawing ◽  
Highly Sensitive ◽  
Favorable Conditions ◽  
Aerobic And Anaerobic

By the employment of methods designed to promote the growth both of aerobic and anaerobic organisms, particularly those belonging to the class of spirochetes, it was possible to obtain a pure culture of a delicate organism, the morphological features of which place it in the genus Leptospira. On three occasions, that is, from three out of eleven cases of yellow fever, the organism was directly cultivated. These three strains were found to induce the characteristic symptoms and lesions when tested on guinea pigs. The organism was designated Leptospira icteroides. Leptospira icteroides was also obtained in pure culture from the blood of guinea pigs which succumbed to infection after being inoculated with the blood or organ emulsions from patients suffering from yellow fever. These cultures also proved to be virulent when tested on susceptible animals. The morphological characteristics and certain biological properties of the organism were considered in detail. It is invisible under translucent illumination and is difficult to stain by most aniline dyes. It is highly sensitive to the presence of bacteria and is rapidly destroyed in a medium in which certain other organisms are present. The presence of blood serum (man, sheep, horse, rabbit, etc.) seems to be essential for its growth. It grows well at a temperature of about 25–26°C. and more quickly at 37°C., though at the latter temperature it dies out within a few weeks. At 25°C. under favorable conditions and in suitable culture media it remains viable for several months without losing its virulence. Leptospira icteroides multiplies by transverse division. The virulence attained by some strains was such that 0.00001 cc. of a culture could induce typical fatal infection in guinea pigs. There exists a considerable variation among guinea pigs in their susceptibility to Leptospira icteroides. The organism is killed within 10 minutes at a temperature of 55°C. and is also destroyed by complete desiccation or freezing and thawing. Bile and bile salts dissolve it in certain concentrations, but not saponin. Leptospira icteroides passes through the pores of Berkefeld filters V and N, and there is a possibility of its having a granular phase of life under certain conditions.

scholarly journals THE PURE CULTIVATION OF SPIROCHÆTA ICTEROHÆMORRHAGIÆ (INADA)

1916 ◽  
Vol 23 (4) ◽  
pp. 557-562 ◽  
Author(s):  
Tetsuta Ito ◽  
Haruichiro Matsuzaki
Keyword(s):  
Causative Agent ◽  
Guinea Pigs ◽  
Culture Media ◽  
Biological Properties ◽  
Facultative Anaerobe ◽  
Fluid Medium ◽  
Weil’S Disease ◽  
Pure Cultures ◽  
Weil's Disease ◽  
Different Strains

Pure cultures of the spirochætal causative agent of the disease known as Weil's disease, or febrile icterus, in Japan, have been obtained by us in a solid, a semisolid, and a fluid medium. The spirochæta thus isolated remains pathogenic for guinea pigs for many generations. Up to the present time we have succeeded through the courtesy of Professor Nagayo, Dr. Konuma, and Dr. Ishihara, in cultivating three different strains. The spirochæta is a facultative anaerobe. The solid and semisolid culture media possess one disadvantage, in that they are opaque on account of the addition of red blood corpuscles; but it is hoped that this drawback may soon be overcome by further studies. We shall report later the results of investigations regarding various questions in immunity as well as further details regarding the biological properties of the spirochæta.

Introduction

1978 ◽  
Vol 36 (3) ◽  
pp. 543-544
Author(s):  
W. Bernard
Keyword(s):  
Molecular Weight ◽  
Electron Microscope ◽  
Nucleic Acid ◽  
Gene Mapping ◽  
Molecular Genetics ◽  
Cell Biology ◽  
Gene Activity ◽  
Close Connection ◽  
Basic Information ◽  
Simple Systems

In comparison to many other fields of ultrastructural research in Cell Biology, the successful exploration of genes and gene activity with the electron microscope in higher organisms is a late conquest. Nucleic acid molecules of Prokaryotes could be successfully visualized already since the early sixties, thanks to the Kleinschmidt spreading technique - and much basic information was obtained concerning the shape, length, molecular weight of viral, mitochondrial and chloroplast nucleic acid. Later, additonal methods revealed denaturation profiles, distinction between single and double strandedness and the use of heteroduplexes-led to gene mapping of relatively simple systems carried out in close connection with other methods of molecular genetics.

Structural integrity after cold storage of human epidermal model in hypothermosol: A correlative ultrastructural and fluorescent assay

1994 ◽  
Vol 52 ◽  
pp. 270-271
Author(s):  
Henry H. Eichelberger ◽  
John G. Baust ◽  
Robert G. Van Buskirk
Keyword(s):  
Cold Storage ◽  
Structural Integrity ◽  
Culture Media ◽  
Cell Structure ◽  
Natural State ◽  
Sodium Cacodylate ◽  
Ruthenium Tetroxide ◽  
Cacodylate Buffer ◽  
Before And After

For research in cell differentiation and in vitro toxicology it is essential to provide a natural state of cell structure as a benchmark for interpreting results. Hypothermosol (Cryomedical Sciences, Rockville, MD) has proven useful in insuring the viability of synthetic human epidermis during cold-storage and in maintaining the epidermis’ ability to continue to differentiate following warming.Human epidermal equivalent, EpiDerm (MatTek Corporation, Ashland, MA) consisting of fully differentiated stratified human epidermal cells were grown on a microporous membrane. EpiDerm samples were fixed before and after cold-storage (4°C) for 5 days in Hypothermosol or skin culture media (MatTek Corporation) and allowed to recover for 7 days at 37°C. EpiDerm samples were fixed 1 hour in 2.5% glutaraldehyde in sodium cacodylate buffer (pH 7.2). A secondary fixation with 0.2% ruthenium tetroxide (Polysciences, Inc., Warrington, PA) in sodium cacodylate was carried out for 3 hours at 4°C. Other samples were similarly fixed, but with 1% Osmium tetroxide in place of ruthenium tetroxide. Samples were dehydrated through a graded acetone series, infiltrated with Spurrs resin (Polysciences Inc.) and polymerized at 70°C.

IMMU-53. STING-ING GLIOBLASTOMA WITH SPHERICAL NUCLEIC ACIDS

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi104-vi105
Author(s):  
Akanksha Mahajan ◽  
Lisa Hurley ◽  
Serena Tommasini-Ghelfi ◽  
Corey Dussold ◽  
Alexander Stegh ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
High Surface ◽  
Biological Properties ◽  
Innate Immune ◽  
Limiting Factors ◽  
Nucleic Acid Delivery ◽  
Sting Pathway ◽  
Spherical Nucleic Acids

Abstract The Stimulator of Interferon Genes (STING) pathway represents a major innate immune sensing mechanism for tumor-derived DNA. Modified cyclic dinucleotides (CDNs) that mimic the endogenous STING ligand cGAMP are currently being explored in patients with solid tumors that are amenable to intratumoral delivery. Inadequate bioavailability and insufficient lipophilicity are limiting factors for clinical CDN development, in particular when consideration is given to systemic administration approaches. We have shown that the formulation of oligonucleotides into Spherical Nucleic Acid (SNA) nanostructures, i.e.,the presentation of oligonucleotides at high density on the surface of nanoparticle cores, lead to biochemical and biological properties that are radically different from those of linear oligonucleotides. First-generation brain-penetrant siRNA-based SNAs (NCT03020017, recurrent GBM) have recently completed early clinical trials. Here, we report the development of a STING-agonistic immunotherapy by targeting cGAS, the sensor of cytosolic dsDNA upstream of STING, with SNAs presenting dsDNA at high surface density. The strategy of using SNAs exploits the ability of cGAS to raise STING responses by delivering dsDNA and inducing the catalytic production of endogenous CDNs. SNA nanostructures carrying a 45bp IFN-simulating dsDNA oligonucleotide, the most commonly used and widely characterized cGAS activator, potently activated the cGAS-STING pathway in vitro and in vivo. In a poorly immunogenic and highly aggressive syngeneic mouse glioma model, in which tumours were well-established, only one dose of intranasal treatment with STING-SNAs decelerated tumour growth, improved survival and importantly, was well-tolerated. Our use of SNAs addresses the challenges of nucleic acid delivery to intracranial tumor sites via intranasal route, exploits the binding of dsDNA molecules on the SNA surface to enhance the formation of a dimeric cGAS:DNA complex and establishes cGAS-agonistic SNAs as a novel class of immune-stimulatory modalities for triggering innate immune responses against tumor.

scholarly journals Effect of kinetin on nucleic acid synthesis in senescing and young leaves in Brassica oleracea L. var. gongylodes L.

2015 ◽  
Vol 44 (4) ◽  
pp. 553-565
Author(s):  
J. Legocka ◽  
A. Szweykowska
Keyword(s):  
Molecular Weight ◽  
Nucleic Acid ◽  
Acid Synthesis ◽  
Inhibitory Effect ◽  
Rrna Synthesis ◽  
Low Molecular Weight ◽  
Chlorophyll B ◽  
Mature Leaves ◽  
Young Leaves ◽  
Brassica Oleracea L

In detached kohlrabi leaves senescing in the dark, the decrease in chlorophyll to was more pronounced than in chlorophyll a. The retardation by kinetin of the chlorophyll loss was also markedly stronger in the case of chlorophyll b. Using the fractionation of nucleic acids on polyacrylamide gels it has been shown that during leaf senescence the level of all RNA species decreased, whereas the amount of DNA was more or less constant. In the presence of kinetin, the loss of RNA was inhibited and the incorporation of precursor into the cytoplasmic rRNA as well as into low molecular weight RNA species was supported. Chloroplast rRNA synthesis has not been detected in mature leaves and kinetin showed no effect in this respect. In young expanding leaves detached and kept in light, the synthesis of cytoplasmic rRNA was strongly stimulated by kinetin, whereas in the case of Chloroplast rRNA only an inhibitory effect of kinetin could be found. The results suggest that the cytokinins are primarily involved in processes of the synthesis of cytoplasmic rRNA and low molecular RNA fractions, and in this way affect the development of plastids, in particular the course of their senescence.

scholarly journals Studies in vivo on the biosynthesis of collagen and elastin in ascorbic acid-deficient guinea pigs. Evidence for the formation and degradation of a partially hydroxylated collagen

1970 ◽  
Vol 119 (3) ◽  
pp. 575-585 ◽  
Author(s):  
M. J. Barnes ◽  
B. J. Constable ◽  
L. F. Morton ◽  
E. Kodicek
Keyword(s):  
Molecular Weight ◽  
Ascorbic Acid ◽  
Guinea Pigs ◽  
Specific Radioactivity ◽  
Rapid Decline ◽  
Time Intervals ◽  
Skin Collagen ◽  
Labelled Compound ◽  
Degree Of Degradation

1. After the administration of l-[G-3H]proline to guinea pigs deprived of ascorbic acid for increasing periods of time, the specific radioactivities of proline and hydroxyproline in skin collagen and aortic elastin were determined at various time-intervals after administration of the labelled compound with a view to studying the formation and degradation of collagen and elastin both deficient in hydroxyproline. 2. As judged from the incorporation of radioactivity into elastin proline, elastin synthesis was not decreased in the ascorbic acid-deficient animals. There was however, a rapid decline in the specific radioactivity of elastin hydroxyproline. The proline/hydroxyproline specific-radioactivity ratio was approx. 1.5:1 after 6 days and 20:1 after 12 days of ascorbic acid deprivation, in contrast with the ratio of 1:1 in controls. The results suggested that the effect of ascorbic acid deficiency on elastin biosynthesis could be regarded as simply an elimination of hydroxylation of elastin proline with the formation and retention of a polymer increasingly deficient in hydroxyproline. 3. Collagen proline and hydroxyproline specific radioactivities were derived from material that was soluble in hot trichloroacetic acid, non-diffusible and collagenase-degradable. In contrast with elastin, there was a rapid decline in the specific radioactivity of proline as well as hydroxyproline in collagen from the ascorbic acid-deficient animals. However, the proline/hydroxyproline specific-radioactivity ratio in all samples from scorbutic animals was consistently slightly above 1:1. The results suggest the appearance in place of collagen, but in rapidly diminishing amounts, of a partially hydroxylated collagen in which the degree of hydroxylation may be decreased only by approx. 10%. 4. Incorporation of radioactivity into the diffusible hydroxyproline in skin remained relatively high despite the rapid decline in the incorporation of radioactivity into collagen. This observation is interpreted as indicative of an increasing degree of degradation of partially hydroxylated collagen to diffusible peptides. An alternative explanation might be that partially hydroxylated peptides are released to an increasing extent from ribosomes before they attain a length at least sufficient to render them non-diffusible. In either case it implies the accumulation in scurvy of low-molecular-weight peptides enriched in proline and deficient in hydroxyproline and could explain the failure to accumulate a high-molecular-weight collagen deficient in hydroxyproline. 5. It is thought, however, that, in addition, an inhibition of ribosomal amino acid incorporation leading to decreased synthesis of partially hydroxylated collagen may also occur, perhaps secondarily to impaired hydroxylation.

INFLUENCE OF THE SULFATION PATTERN ON CERTAIN BIOLOGICAL PROPERTIES OF GALACTOSAMINOGLYCANS

1987 ◽  
Author(s):  
B Casu ◽  
L Marchese ◽  
A Naggi ◽  
G Torri ◽  
J Fareed ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Dermatan Sulfate ◽  
Anticoagulant Activity ◽  
Chlorosulfonic Acid ◽  
Biological Properties ◽  
Sulfated Polysaccharides ◽  
Low Molecular Weight ◽  
Antithrombotic Activity

In order to investigate the influence of charge distribution and chain length on the biological properties of sulfated polysaccharides, additional sulfate groups were introduced into the galactosaminoglycans, chondriotin sulfate and dermatan sulfate. Using a flexible method (with sulfuric acid and chlorosulfonic acid) for concurrent sulfation and controlled depolymerization, numerous products were obtained and characterized by chemical, enzymatic and nuclear magnetic resonance spectroscopic methods. The biologic actions of these products were profiled in both in vitro and in vivo assays for antithrombotic activity. Despite a weaker in vitro anticoagulant activity, low molecular weight over sulfated galactosaminoglycans produced significant dose-dependent antithrombotic actions in animal models which were similar to the actions observed with oversulfated low molecular weight heparins. These results suggest that a significant antithrombotic activity can be elicited through non-specific interactions of polysulfates with cellular and plasma components, and that clusters of sulfate groups such as the 4-6 disulfate group on D-galactosaminoglycan residues may be important for these interactions. Furthermore, these results, also suggest that supersulfation of glycosaminogly-cans results in products with biologic activity distinct from the native material.

scholarly journals Gas Chromatography-Mass Spectrometry (GC-MS) Analysis of Essential Oils from AgNPs and AuNPs Elicited Lavandula angustifolia In Vitro Cultures

Molecules ◽  
2019 ◽  
Vol 24 (3) ◽  
pp. 606 ◽  
Author(s):  
Aneta Wesołowska ◽  
Paula Jadczak ◽  
Danuta Kulpa ◽  
Włodzimierz Przewodowski
Keyword(s):  
Molecular Weight ◽  
Silver Nanoparticles ◽  
Essential Oils ◽  
Culture Media ◽  
In Vitro Cultures ◽  
Gas Chromatography Mass Spectrometry ◽  
Lavandula Angustifolia ◽  
Gold And Silver Nanoparticles ◽  
The Media

The aim of this study was to determine how the addition of gold and silver nanoparticles to culture media affects the composition of essential oils extracted from Lavandula angustifolia propagated on MS media with the addition of 10 and 50 mg·dm−3 of gold (24.2 ± 2.4 nm) and silver (27.5 ± 4.8 nm) nanocolloids. The oil extracted from the lavender tissues propagated on the medium with 10 mg·dm−3 AgNPs (silver nanoparticles) differed the most with respect to the control; oil-10 compounds were not found at all, and 13 others were detected which were not present in the control oil. The addition of AuNPs (gold nanoparticles) and AgNPs to the media resulted in a decrease of lower molecular weight compounds (e.g., α- and β-pinene, camphene, δ-3-carene, p-cymene, 1,8-cineole, trans-pinocarveol, camphoriborneol), which were replaced by those of a higher molecular weight (τ- and α-cadinol 9-cedranone, cadalene, α-bisabolol, cis-14-nor-muurol-5-en-4-one, (E,E)-farnesol).

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