scholarly journals In Vitro Preparation And Testing Of Anti-Salmonella Vaccine Against Abortion In Sheep

2017 ◽  
Vol 74 (1) ◽  
pp. 26
Author(s):  
Flore CHIRILA ◽  
George Cosmin NADAS ◽  
Sorin RAPUNTEAN ◽  
Cosmina Maria BOUARI ◽  
Septimiu TOADER ◽  
...  
Keyword(s):  
Cell Walls ◽  
Culture Supernatant ◽  
Serum Antibody ◽  
Brain Heart Infusion ◽  
Broth Culture ◽  
Millipore Filter ◽  
Serum Titer ◽  
Sterility Control

Abstract: In March 2016, the microbiology laboratory of the Faculty of Veterinary Medicine in Cluj-Napoca, 3 type of antisalmonella vaccine for sheep were prepared. For type 1, 24 hours salmonella on brain heart infusion (BHI) broth culture, heat inactivated for 1 hour at 60°C, then with formaldehyde in a concentration of 3 ‰. Variant 2 - the culture supernatant obtained on solid BHI medium, washed with PBS, frozen-thawed 6 times, centrifuged at 4000 rpm for 15 minutes, filtered through 0.2 μm Millipore filter Orange Scientific. Variant 3 - suspension of cell walls remaining after centrifugation suspended in PBS and inactivated with formaldehyde.After the bacterial and fungal sterility control, the three types of vaccine were administered in rabbits by the subcutaneous route at a dose of 1 ml/individual along with 1 ml of plant extract adjuvant (decoction of Ewernia prunastri) sterilized by filtration.There have been two booster inoculations of the initial administration, 7 and 14 days after. Before each dose of vaccine, blood was sampled from the marginal auricular vein in order to control the immunogenicity by anti-somatic ˮOˮ serum antibody (Ab) titration using slow microplate agglutination test (Widal reaction). After three inoculations with the vaccine variant 1, Ab serum titer reached 1/128, and in types 2 and 3, 1/512 after 2 inoculations, decreasing to 1/256 after the second booster administered with no immunomodulator.

scholarly journals PREPARATION AND ANTIGENICITY OF M PROTEIN RELEASED FROM GROUP A, TYPE 1 STREPTOCOCCAL CELL WALLS BY PHAGE-ASSOCIATED LYSIN

1960 ◽  
Vol 112 (1) ◽  
pp. 77-96 ◽  
Author(s):  
Fred S. Kantor ◽  
Roger M. Cole
Keyword(s):  
Cell Walls ◽  
Serum Antibody ◽  
Intradermal Injection ◽  
Exchange Chromatography ◽  
M Protein ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Bactericidal Antibody ◽  
Group A

The lysis of Group A type 1 streptococcal cell walls by phage-associated lysin has been described. In the preparation of lysin, a new semisynthetic non-protein media to support growth of the propagating strain of Group C streptococci was employed. Following lysis of the cell walls, the resulting digest was partially purified by ion-exchange chromatography and ammonium sulfate precipitation. In addition to M protein, the resulting preparation (called lysin M protein) contained the group-specific carbohydrate and the T protein—but did not contain antigens, detectable by precipitin tests, which cross-reacted with absorbed heterologous group or type antisera. Capillary precipitin reactions between the lysin M protein and type-specific antiserum did not occur in the presence of high ionic strength buffers; these buffers did not similarly affect precipitin reactions of acid M protein. Type 1 lysin M protein is shown to be a good antigen. A total of 1.5 mg. injected intradermally in saline produced bactericidal antibody in eight of nine rabbits; when injected in adjuvant in one rabbit, protective serum antibodies developed. Streptococci grown in sera from seven of nine rabbits immunized with lysin M protein demonstrated significantly longer chains than when grown in normal rabbit serum. Antibody as demonstrated by each of these three tests was shown to be type-specific. No local or systemic toxicity was noted following intradermal injection in rabbits of lysin M protein.

scholarly journals Inhibitory Effects of Commercial and Enriched Green Tea Extracts on the Growth of Brochothrix thermosphacta, Pseudomonas putida and Escherichia coli

2012 ◽  
Vol 2 (1) ◽  
pp. 1 ◽  
Author(s):  
Elodie Rozoy ◽  
Laurent Bazinet ◽  
Monica Araya-Farias ◽  
Anthony Guernec ◽  
Linda Saucier
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Putida ◽  
Green Tea ◽  
Antimicrobial Properties ◽  
Epigallocatechin Gallate ◽  
Brain Heart Infusion ◽  
Broth Culture ◽  
Meat Spoilage ◽  
Brochothrix Thermosphacta

<p>The major catechin found in green tea, called epigallocatechin gallate (EGCG), have been reported to have antimicrobial properties. In this study, we examined <em>in vitro </em>the antimicrobial effects of a commercial green tea extract sold in a capsule form, and two prepared green tea extracts enriched in catechins against <em>Brochothrix thermosphacta</em>, <em>Pseudomonsas putida</em> and <em>Escherichia coli</em> which have been associated with meat spoilage. The antimicrobial activity of the different tea extracts was evaluated by Spot-On-Lawn and Well Diffusion assays and the Minimum Inhibitory Concentration (MIC) was also determined in Brain Heart Infusion broth. The three methods used showed an inhibition of <em>Brochothrix thermosphacta</em>, whereas the inhibition of <em>Pseudomonas putida</em> and <em>Escherichia coli</em> was only detected with the MIC assay. The determination of the MIC in broth culture appeared to be the most reliable method to determine the inhibitory activity of catechin compounds. <strong></strong></p>

scholarly journals Enhanced Murine Antigen-Specific Gamma Interferon and Immunoglobulin G2a Responses by Using Mycobacterial ESAT-6 Sequences in DNA Vaccines

2003 ◽  
Vol 71 (4) ◽  
pp. 2239-2243 ◽  
Author(s):  
F. Chris Minion ◽  
Sreekumar A. Menon ◽  
Gregory G. Mahairas ◽  
M. J. Wannemuehler
Keyword(s):  
Immune Responses ◽  
Dna Vaccines ◽  
Serum Antibody ◽  
Interleukin 10 ◽  
Gamma Interferon ◽  
Viral Pathogens ◽  
Ifn Γ ◽  
Antibody Levels

ABSTRACT The Mycobacterium tuberculosis protein ESAT-6 has unusual immune stimulating activities, has been implicated in the recall of long-lived immunity, and induces protection against tuberculosis in mice. For many diseases caused by bacterial or viral pathogens, a strong cell-mediated immune (i.e., type 1) response is often required for recovery or protection. Therefore, it is important to design immunization regimens that induce agent-specific type 1 immunity. We have shown in previous studies that ESAT-6 could enhance antigen-specific type 1 immune responses in BALB/c mice against a second antigen when presented as a purified fusion protein. It was also of interest to determine if ESAT-6 could enhance the type 1 response against a second antigen beyond that afforded by DNA vaccination through CpG motifs. This was tested by using gene fusions of ESAT-6 and the Mycoplasma hyopneumoniae surface antigen P71. Modified P71 gene sequences were cloned with or without ESAT-6 sequences into a DNA vaccine vector and were used to immunize mice. Splenic lymphocytes from vaccinated mice were tested for gamma interferon (IFN-γ) and interleukin-10 (IL-10) secretion. Serum antibodies were examined for P71 antigen-specific isotype responses. When stimulated in vitro with purified P71 antigen, splenocytes from the ESAT-6:P71 vaccinates secreted higher levels of IFN-γ and lower levels of IL-10 compared to those of vaccinates receiving the P71 construct alone. Furthermore, the immunoglobulin G2a serum antibody levels were significantly higher in the ESAT-6:P71 vaccinates compared to those of the vaccinates receiving P71 alone. In conclusion, ESAT-6 was shown to enhance antigen-specific type 1 immune responses in BALB/c mice when used in DNA vaccines.

Apoptosis and proliferation ofperipheral blood T-cells as alternative processes in pathogenesis of diabetes mellitus type 1

2019 ◽  
Vol 1 (4) ◽  
pp. 16-20 ◽  
Author(s):  
A. V. Lugovaya ◽  
N. M. Kalinina ◽  
V. Ph. Mitreikin ◽  
Yu. P. Kovaltchuk ◽  
A. V. Artyomova ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
T Cells ◽  
High Risk ◽  
Peripheral Blood ◽  
Cellular Response ◽  
Proliferative Potential ◽  
Autoreactive T Cells ◽  
Alternative Processes

Apoptosis, along with proliferation, is a form of lymphocyte response to activating stimuli. In the early stages of cell differentiation, the apoptotic response prevails and it results to the formation of tolerance to inductor antigen. Mature lymphocytes proliferate in response to stimulation and it means the initial stage in the development of the immune response. Since in this case apoptosis and proliferation acts as alternative processes, their ratio can serve as a measure of the effectiveness of the cellular response to activating signals. The resistance of autoreactive T-cells to apoptosis is the main key point in the development of type 1 diabetes mellitus (T1DM). Autoreactive T-cells migrates from the bloodstream to the islet tissue of the pancreas and take an active part in b cells destruction. The resistance of autoreactive effector T-cells to apoptosis may suggest their high proliferative potential. Therefore, the comparative evaluation of apoptosis and proliferation of peripheral blood lymphocytes can give a more complete picture of their functional state and thus will help to reveal the causes of ineffective peripheral blood T-ceiis apoptosis in patients with T1DM and will help to understand more deeply the pathogenesis of the disease. in this article, the features of proliferative response of peripheral blood T-cells in patients with T1DM and in individuals with high risk of developing T1DM have been studied. Apoptosis of T-cell subpopulations has been investigated. The correlation between the apoptotic markers and the intensity of spontaneous and activation- induced in vitro T-cells proliferation of was revealed. it was determined, that autoreactive peripheral blood T-cells were resistant to apoptosis and demonstrated the increased proliferative potential in patients with T1DM and in individuals with high risk of developing T1DM.

DETERMINATION OF BLOOD CORTICOIDS USING THE IN VITRO RESIN UPTAKE OF 3H-PREDNISOLONE

1968 ◽  
Vol 57 (1) ◽  
pp. 23-32 ◽  
Author(s):  
Hironori Nakajima ◽  
Mitsunori Murala ◽  
Masumitsu Nakata ◽  
Takeshi Naruse ◽  
Seiji Kubo
Keyword(s):  
Thyroid Function Test ◽  
Normal Subjects ◽  
Adrenal Function ◽  
Function Test ◽  
Before And After ◽  
Adrenal Response ◽  
Suppression Test

ABSTRACT The in vitro resin uptake of 3H-prednisolone was used for the determination of blood cortisol after addition of radioactive prednisolone followed by Amberlite CG 400 Type 1 to the test serum, and incubation of the mixture. The radioactivity of the supernatant was compared before and after the addition of the resin. The principle of this method is similar to that of the 131I-triiodothyronine resin uptake for the thyroid function test. The tests for the specificity, reproducibility and sensitivity gave satisfactory results. The mean basal value ± SD of the 3H-prednisolone resin uptake was 35.3 ± 9.2% in normal subjects, and 27.1 ± 4.8% in pregnant women. This method was valid in various adrenal function tests, i. e. the adrenal circadian rhythm, corticotrophin (ACTH) test, dexamethasone suppression test and the adrenal response to lysine-8-vasopressin. It proved to be a sensitive indicator of the adrenal function. These results suggest that this method should be useful for a routine adrenal function test.

An In Vitro Analysis of Sodium Hypochlorite Decontamination for the Reuse of Implant Healing Abutments

2020 ◽  
Author(s):  
Ahmad Almehmadi
Keyword(s):  
Sodium Hypochlorite ◽  
Bacterial Culture ◽  
Brain Heart Infusion ◽  
In Vitro Study ◽  
Vitro Study ◽  
Sem Images ◽  
Streptococcus Sanguis ◽  
Implant Dentistry ◽  
Vitro Analysis

Abstract The re-use of healing abutments (HAs) has become common practice in implant dentistry for economic concerns and the aim of this in-vitro study was to assess the effect of sodium hypochlorite (NaOCl) in decontamination of HAs. 122 HAs (Used and sterilized n=107; New n=15) were procured from 3 centers, of which 3 samples were discarded due to perforation in sterilization pouch.  For sterility assessment, the used HAs (n=80) were cultured in Brain Heart Infusion Broth (BHI) and Potato Dextrose Agar (PDA), bacterial isolates were identified in 7 samples. Also, 24 used HAs were stained with Phloxine B, photographed and compared to new HAs (n=5). Scanning electron microscope (SEM) assessed the differences between the two sets of HAs, following which the 7 contaminated HAs along with 24 used HAs from staining experiment (Total=31) were subsequently treated with sodium hypochlorite (NaOCl) and SEM images were observed. About 8.75% of HAs tested positive in bacterial culture; Streptococcus sanguis, Dermabacter hominis, Staphylococcus haemolyticus, and Aspergillus species were isolated. Phloxine B staining was positive for used and sterilized HAs when compared to controls. The SEM images revealed deposits in the used HAs and although treatment with NaOCl eliminated the contamination of cultured HAs, the SEM showed visible debris in the HA thread region. This in-vitro study concluded that SEM images showed debris in used HAs at screw-hole and thread regions even though they tested negative in bacterial culture. The treatment with NaOCl of used HAs showed no bacterial contamination but the debris was observed in SEM images. Future studies on the chemical composition, biological implications, and clinical influence is warranted before considering the reuse of HAs.

Methods of experimental polyploidization and their use in apple breeding

2020 ◽  
Vol 62 ◽  
pp. 85-90
Author(s):  
L. V. Tashmatova ◽  
O. V. Matsneva ◽  
T. M. Khromova ◽  
V. V. Shakhov
Keyword(s):  
Exposure Time ◽  
Cell Walls ◽  
Prolonged Exposure ◽  
Ornamental Plants ◽  
Colchicine Treatment ◽  
Apple Trees ◽  
Open Ground ◽  
High Concentrations ◽  
Diploid Gametes

The article presents methods of experimental polyploidy of fruit, berry and ornamental plants. The purpose of this review is to highlight the problems and prospects of polyploidization of plants in the open ground and in vitro culture and the possibility of their application for apple trees. For the purpose of obtaining apple tetraploids as donors of diploid gametes, seed seedlings were treated with a solution of colchicine in concentrations of 0.1-0.4 % for 24 and 48 hours. Colchicine concentrations of 0.3 % and 0.4 % at 48 hours of treatment had a detrimental eff ect on their development. As a result, tetraploids and chimeras were obtained from seeds from free pollination of the varieties Orlik, Svezhest, Kandil Orlovsky, as well as from seeds obtained from crossing the varieties Svezhest×Bolotovskoe, Moskovskoe Оzherel’e×Imrus, Girlyanda×Venyaminovskoe. The optimal concentration of colchicine was 0.1 %. Methods of colchicine treatment have been studied: 1) adding to the nutrient medium, colchicine concentration: 0.01%, 0.02%, exposure time 24h-19 days; 2) applying amitotic solution to the growth point, colchicine concentration: 0.1 %, 0.2 %, exposure time 24h-7 days. To increase the penetration of colchicine through the cell walls, a 0.1 % dimexide solution was used. Studies have shown that high concentrations and prolonged exposure to colchicine reduce the viability of explants.

Antifungal Proteins from Plant Latex

2020 ◽  
Vol 21 (5) ◽  
pp. 497-506
Author(s):  
Mayck Silva Barbosa ◽  
Bruna da Silva Souza ◽  
Ana Clara Silva Sales ◽  
Jhoana D’arc Lopes de Sousa ◽  
Francisca Dayane Soares da Silva ◽  
...  
Keyword(s):  
Cell Walls ◽  
Defense Mechanisms ◽  
Hydrolytic Enzymes ◽  
Fungal Species ◽  
Biologically Active ◽  
Protein Classification ◽  
Fungal Cell ◽  
Antifungal Proteins ◽  
Plant Latex

Latex, a milky fluid found in several plants, is widely used for many purposes, and its proteins have been investigated by researchers. Many studies have shown that latex produced by some plant species is a natural source of biologically active compounds, and many of the hydrolytic enzymes are related to health benefits. Research on the characterization and industrial and pharmaceutical utility of latex has progressed in recent years. Latex proteins are associated with plants’ defense mechanisms, against attacks by fungi. In this respect, there are several biotechnological applications of antifungal proteins. Some findings reveal that antifungal proteins inhibit fungi by interrupting the synthesis of fungal cell walls or rupturing the membrane. Moreover, both phytopathogenic and clinical fungal strains are susceptible to latex proteins. The present review describes some important features of proteins isolated from plant latex which presented in vitro antifungal activities: protein classification, function, molecular weight, isoelectric point, as well as the fungal species that are inhibited by them. We also discuss their mechanisms of action.

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