PSEUDOURIDINE FROM CULTURE MEDIA OF AGROBACTERIUM TUMEFACIENS

1964 ◽  
Vol 10 (6) ◽  
pp. 867-875 ◽  
Author(s):  
T. Suzuki ◽  
R. M. Hochster

Cultures of Agrobacterium tumefaciens grown on a synthetic medium containing glucose plus a pyrimidine produced a pyrimidine riboside with unusual chemical properties. This substance was isolated in crystalline form and identified as 5-ribosyluracil (pseudouridine) on the basis of data obtained from ion exchange and paper chromatography, electrophoresis, ultraviolet and infrared spectrophotometry, color reactions, melting point, elementary analysis, hydrazinolysis, and periodate oxidation followed by bromine treatment.

Tuberculosis (TB) is one of the most important zoonotic bacterial diseases. A huge economic loss which could be direct or indirect are associated with the disease. Currently, the primary methods used for detection of TB in humans and cattle include the measurement of a delayed type hypersensitivity to purified protein derivative (PPD). So, the need for preparation of purified PPD with adequate properties and increasing the final PPD yield with decreasing the time of tuberculin production has stimulated the interest in the development of its preparation. Our study was performed to compare between the standard and modified media for improving tuberculin production. Middle brook 7H10 agar medium was used as a modified basic medium for mycobacterial growth, followed by cultivation of mycobacteria on Middle brook 7H9 broth medium. For the production, strains were inoculated onto the culture medium (Dorest Henly synthetic medium). Other steps for tuberculin production was done according to standard Weighbridge protocol. The results demonstrated that the using of both Middle brook 7H10 agar and Middle brook 7H9 broth instead of Lowenstein-Jensen (LJ) and glycerin broth media which used in currently produced tuberculin, have better physical and chemical properties. In addition, reducing the time required for production by accelerating the time of microbial growth. Also, it was found that the tuberculin produced using modified media was slightly more potent or the same as currently tuberculin produced. So, both Middle brook 7H10 agar and Middle brook 7H9 broth media are recommended for production of tuberculin saving time and increasing potency of the product but more investigation was recommended for estimation types of protein present in both locally prepared and modified tuberculin.


1932 ◽  
Vol 7 (3) ◽  
pp. 285-292 ◽  
Author(s):  
J. Ansel Anderson

Khapli wheat leaves contain a very small quantity of a trihydroxydimethoxyflavone, tricin. Marquis yields a trace of the same compound. These two varieties also contain water-soluble coloring matters which are apparently glucosides of tricin or of a closely related compound. Dyeing tests carried out with six other wheat varieties indicate that all contain essentially the same coloring matters.Methylation of tricin and hydrolysis of the resulting O-trimethyltricin yielded 3,4,5-trimethoxybenzoic acid and 2-hydroxy-4,6-dimethoxyacetophenone. It was therefore assumed that tricetin, the pentahydroxyflavone derived from tricin, was 5,7,3′,4′,5′-pentahydroxyflavone. This was synthesized from 3,4,5-trimethoxybenzoic acid and phloracetophenone by the Allan-Robinson method. Mixed melting-point determinations showed that its O-pentamethyl and O-penta-acetyl derivatives are identical with the corresponding derivatives of tricetin. The dyeing properties and color reactions of synthetic and natural tricetin are identical and are in fair agreement with those described by Badhwar, Kang and Ventkataraman (3, p. 1111) who recently reported the synthesis of the same compound.


Vaccines ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 40
Author(s):  
Sandra Guallar-Garrido ◽  
Farners Almiñana-Rapún ◽  
Víctor Campo-Pérez ◽  
Eduard Torrents ◽  
Marina Luquin ◽  
...  

Mycobacterium bovis bacillus Calmette-Guérin (BCG) efficacy as an immunotherapy tool can be influenced by the genetic background or immune status of the treated population and by the BCG substrain used. BCG comprises several substrains with genetic differences that elicit diverse phenotypic characteristics. Moreover, modifications of phenotypic characteristics can be influenced by culture conditions. However, several culture media formulations are used worldwide to produce BCG. To elucidate the influence of growth conditions on BCG characteristics, five different substrains were grown on two culture media, and the lipidic profile and physico-chemical properties were evaluated. Our results show that each BCG substrain displays a variety of lipidic profiles on the outermost surface depending on the growth conditions. These modifications lead to a breadth of hydrophobicity patterns and a different ability to reduce neutral red dye within the same BCG substrain, suggesting the influence of BCG growth conditions on the interaction between BCG cells and host cells.


2014 ◽  
Vol 26 (1) ◽  
pp. 154 ◽  
Author(s):  
D. Moreno ◽  
A. Neira ◽  
L. Dubreil ◽  
L. Liegeois ◽  
S. Destrumelle ◽  
...  

In the majority of media for embryo culture, 2 of typical components used are FCS or BSA; however, the presence of FCS in the culture medium has been shown to have a negative effect on embryo quality and the use of animal-derived proteins in culture media increases the risks of disease transmission through in vitro embryo production. The aim of this study was to develop an in vitro embryo culture medium free from FCS and BSA, but with the addition of various growth factors and cytokines (GF-CYK: IGF-I, IGF-II, bFGF, LIF, GM-CSF) 50 ng mL–1 and (TGF-β1) 100 ng mL–1 supplemented with hyaluronan (HA) and recombinant albumin (RA). Bovine oocytes (n = 1043, 6 replicates) from abattoir ovaries were matured in TCM-199 medium with 60 μg mL–1 penicillin, 60 μg mL–1 streptomycin, and 10 ng mL–1 EGF for 24 h at 39°C and 5% CO2 in humidified air. Afterward, the oocytes were fertilized in IVF-TALP medium with 6 mg mL–1 fatty acid-free BSA and 1.7 IU mL–1 heparin for 18 h under the same conditions. After fertilization, presumptive zygotes were divided into two groups and cultured in 30 μL droplets of SOF supplemented with (1) 0.4% BSA + 5 μg mL–1 insulin, 5 μg mL–1 transferrin, and 5 ng mL–1 selenium (ITS) as a control; or (2) GF-CYK + 0.5 mg mL–1 HA + 0.15% RA (M1). Droplets were preserved under mineral oil in a humidified atmosphere of 5% CO2, 5% O2, and 90% N2 at 39°C. Blastocyst development and blastocyst diameter was observed at 7 and 8 days post-fertilization (dpf). Developmental and diameter data were analysed using the Wilcoxon test by using R software. The blastocyst rates were not significantly different between the control and M1 medium: at 7 dpf (22.9% ± 4.8 and 30.2% ± 3.0), and at 8 dpf (29.6% ± 5.1 and 37.4% ± 2.0 respectively; P > 0.05). The blastocyst diameter obtained with the M1 medium was significantly greater (P < 0.05) than that of the control at 7 dpf (173.3 μm ± 4.9 and 157.2 μm ± 4.1, respectively); however, no significant differences were observed at 8 dpf (190.3 μm ± 5.2 and 179.7 μm ± 5.3, respectively). In conclusion, the FCS- and BSA-free medium with GF-CYK, HA, and RA (M1) showed a comparable development rate to the control medium at 7 and 8 dpf. These growth factors and cytokines in association with hyaluronan and recombinant albumin have a synergistic action by promoting an increase in the blastocyst diameter at 7 dpf. This is fully synthetic method of embryo culture; it presents a valuable tool to reduce the risks of disease transmission via embryo transfer.


Hernia ◽  
2020 ◽  
Vol 24 (6) ◽  
pp. 1345-1359 ◽  
Author(s):  
Reiko Wiessner ◽  
R. Lorenz ◽  
A. Gehring ◽  
T. Kleber ◽  
C. Benz ◽  
...  

Abstract Introduction In Africa and other Low Resource Settings (LRS), the guideline-based and thus in most cases mesh-based treatment of inguinal hernias is only feasible to a very limited extent. This has led to an increased use of low cost meshes (LCMs, mostly mosquito meshes) for patients in LRS. Most of the LCMs used are made of polyethylene or polyester, which must be sterilized before use. The aim of our investigations was to determine changes in the biocompatibility of fibroblasts as well as mechanical and chemical properties of LCMs after steam sterilization. Material and methods Two large-pored LCMs made of polyester and polyethylene in a size of 11 x 6 cm were cut and steam sterilized at 100, 121 and 134 °C. These probes and non-sterile meshes were then subjected to mechanical tensile tests in vertical and horizontal tension, chemical analyses and biocompatibility tests with human fibroblasts. All meshes were examined by stereomicroscopy, scanning electron microscopy (SEM), LDH (cytotoxicity) measurement, viability testing, pH, lactate and glycolysis determination. Results Even macroscopically, polyethylene LCMs showed massive shrinkage after steam sterilization, especially at 121 and 134 °C. While polyester meshes showed no significant changes after sterilization with regard to deformation and damage as well as tensile force and stiffness, only the unsterile polyethylene mesh and the mesh sterilized at 100 °C could be tested mechanically due to the shrinkage of the other specimen. For these meshes the tensile forces were about four times higher than for polyester LCMs. Chemical analysis showed that the typical melting point of polyester LCMs was between 254 and 269 °C. Contrary to the specifications, the polyethylene LCM did not consist of low-density polyethylene, but rather high-density polyethylene and therefore had a melting point of 137 °C, so that the marked shrinkage described above occurred. Stereomicroscopy confirmed the shrinkage of polyethylene LCMs already after sterilization at 100 °C in contrast to polyester LCMs. Surprisingly, cytotoxicity (LDH measurement) was lowest for both non-sterile LCMs, while polyethylene LCMs sterilized at 100 and 121 °C in particular showed a significant increase in cytotoxicity 48 hours after incubation with fibroblasts. Glucose metabolism showed no significant changes between sterile and non-sterile polyethylene and polyester LCMs. Conclusion The process of steam sterilization significantly alters mechanical and structural properties of synthetic hernia mesh implants. Our findings do not support a use of low-cost meshes because of their unpredictable properties after steam sterilization.


2012 ◽  
Vol 260-261 ◽  
pp. 593-597
Author(s):  
Veronika Sassmanová ◽  
Zuzana Poláčková ◽  
Jaroslav Frantík ◽  
Stanislav Honus ◽  
Dagmar Juchelková

The article deals with the identification of a chemical structure of pyrolysis liquid occurring by the thermal degradation of tyre debris within the temperature interval from 500°C to 650°C and the temperature increase steps of 50°C in individual experiments and with the use of analytical methods GC/MS, FT-IR and the elementary analysis. The measured FT-IR spectra indicate the representation of functional groups included in measured samples. They could be used for the determination of basic chemical properties. There were changes in the chemical composition monitored as well as the creation of functional groups in the pyrolysis liquid on the basis of FT-IR spectra and GS/MS outcomes.


2015 ◽  
Vol 8 (1) ◽  
pp. 1
Author(s):  
Athina Mardhatillah ◽  
Mutakin Mutakin ◽  
Jutti Levita

Dehydrodiisoeugenol (DDIE) synthesis has been performed by modifying a method recommended by Leopold with a different ratio of isoeugenol and FeCl<sub>3</sub> (1.9:1). FeCl<sub>3</sub> was chosen as catalyst due to its efficiency and environment-friendly property. This modification yielded 22.93 % of product. The product, a white crystalline form, was characterized using thin layer chromatography, melting point, UV, IR, HRMS, and NMR spectroscopy, as well as HPLC, employing pure DDIE as the standard. TLC chromatogram showed Rf 0.32 using n-hexane/ethyl acetate (8:2). The crystals melted at 138-139 <sup>o</sup>C, while its UV maximum was detected at l 273 nm. IR spectrum showed a specific broad O-H stretch at 3437.15 cm<sup>-1</sup>, C-H aromatic and C-H alkene at 3163.26 and 3024.38 cm<sup>-1</sup>, C-H alkyl stretch at 2951.09 and 2927.94 cm<sup>-1</sup>. An overtone peak of aromatic was detected at 2100 to 1700 cm<sup>-1</sup>. C-O peak was detected at 1126.43 cm<sup>-1</sup>. HPLC showed that this compound was eluted at 11.886 minutes after it was injected to a C18 column 250 x 4 mm using a mixture of methanol and double distilled water (73:27) for mobile phase. HRMS spectra predicted that the molecular structure is C<sub>20</sub>H<sub>22</sub>O<sub>4</sub> as showed by abundance peak at <em>m/z </em>327.1595 of [M+H]<sup>+</sup>. <sup>1</sup>H-NMR and <sup>13</sup>C-NMR indicated that the synthesized compound contains 13 types of proton and 20 types of carbon. Herein we reported that white needle-like crystals of DDIE using FeCl<sub>3</sub> as catalyst had been synthesized, moreover the decreasing of the catalyst reduced the yield of the product.


2015 ◽  
Vol 44 (17) ◽  
pp. 6287-6305 ◽  
Author(s):  
Thomas L. Moore ◽  
Laura Rodriguez-Lorenzo ◽  
Vera Hirsch ◽  
Sandor Balog ◽  
Dominic Urban ◽  
...  

This review discusses nanoparticle colloidal stability in biological media in an attempt to shed light on the difficulty correlating nanoparticle physico-chemical properties and biological fate.


Although the substance called Natrolite by Mr. Klaproth has lately been found, under a crystalline form, perfectly similar to that of mesotype, M. Haüy has not thought himself warranted, from this circumstance, to consider these two bodies as of the same species, on account of the difference of their chemical composition; because, according to the analysis of M. Vauquelin, there was not observed to be any soda in zeolite, but, on the contrary, a considerable quantity of lime, which is not a constituent part of natrolite. Mr. Smithson, on the contrary, is inclined to consider them as the same substance, by the agreement of their chemical properties, as well as crystallographical form. Although he had, many years since, found that the zeolite of Staffa contained soda, as has since been observed by Dr. Hutton and Dr. Kennedy, he still felt uncertain whether these were of the same kind as those analysed by M. Vauquelin: but having lately received from M. Haüy a cluster of mesotype in rectangular prisms, terminated by a quadrangular pyramid, Mr. Smithson took this opportunity of ascertaining whether this substance and natrolite did or did not differ in their composition. The method pursued by the author in the analysis of zeolite was first to expose ten grains of it to a strong heat, for the purpose of ascertaining the loss; and he found it to be ·95 grain. An equal quantity was then dissolved in muriatic acid; and after the solution had been evaporated to dryness, the residuum was exposed to a red heat. Water was then poured upon it, and dissolved a portion, which, upon evaporation, was found to be muriate of soda, weighing 3·15 grains. From the solution of this salt, neither carbonate of ammonia nor oxalic acid occasioned any precipitate; by which it appeared that this zeolite contained no lime.


2018 ◽  
pp. 38-47
Author(s):  
O. A. Bihdan ◽  
V. V. Parchenko

There are engaged scientists practically from all over the world to the 1,2,4-triazole system research but the greatest attention deserve the achievements of domestic scientists who managed from the very beginning to register new original medicines in Ukraine. There is no doubt that the original attempt of combining various typical fragments with the nucleus of 1,2,4-triazole is gaining popularity among researchers. The purpose of our work was to synthesized and investigate a row of new 5-(3-fluorophenyl)-4-methyl-1,2,4-triazole-3-thiols by a Sulfur atom, to study the physical-chemical properties of previously unsubscribed compounds. The chemical names of the compounds are given in accordance with the IUPAC nomenclature (1979) and the IUPAC recommendations (1993). Investigations of the physical-chemical properties of the obtained compounds were carried out according to the methods which are given in the State Pharmacopoeia of Ukraine. The melting point was determined on an automatic device for determining the melting point OptiMelt Stanford Research Systems MPA100 (USA). The elemental structure of the new compounds was established on the element analyzer Elementar Vario L cube (CHNS) (standard - sulfanilamide). The data of elemental analysis correspond to the calculated. The 1Н NMR-spectra of the compounds were recorded using a «Mercury 400» spectrometer, the solvent DMSO-d6, the internal standard – tetramethylsilane (TMS) and decrypted using the SpinWorks computer program. Chromate-mass spectra were recorded on an Agilent 6890N/5973N/FID spectrometer produced by Agilent Technologies with a Dean microflow switch. By the experiment results there were obtained eleven previously unidentified individual compounds light yellow and white color which are soluble in organic solvents and insoluble in water. The crystallization process was carried out in isopropanol. Using a complex approach with the use of modern physical-chemical methods of analysis we confirmed the structure of 5-(3-fluorophenyl)-4-methyl-1,2,4-triazole-3-thiol, 3-alkylthio-5-(3-fluorophenyl)-4-methyl-1,2,4-triazole, 1,4-bis((5-(3-fluorophenyl)-4-methyl-1,2,4-triazole-3-yl)thio)propane and 1,4-bis((5-(3-fluorophenyl)-4-methyl-1,2,4-triazole-3-yl)thio)butane. The individuality of the synthesized compounds was confirmed chromatographically, the constants peaks of the pseudo molecular ions of the synthesized compounds coincide with the calculated values of the molecular masses. We first synthesized 5-(3-fluorophenyl)-4-methyl-1,2,4-triazol-3-thiol which was further used as a starting material in the synthesis of new 3-alkylthio-5-(3-fluorophenyl)-4-methyl-1,2,4-triazole, 1,4-bis((5-(3-fluorophenyl)-4-methyl-1,2,4-triazol-3-yl)thio)propane and 1,4-bis((5-(3-fluorophenyl)-4-methyl-1,2,4-triazol-3-yl)thio)butane. Complex physical-chemical methods of analysis confirmed the structure of synthesized compounds.


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