Antigenic analysis of Pasteurella multocida (serotype 1) by crossed immunoelectrophoresis: characterization of cytoplasmic and cell envelope associated antigens

1980 ◽  
Vol 26 (6) ◽  
pp. 676-689
Author(s):  
J. L. Bhasin ◽  
L. Lapointe-Shaw
Keyword(s):  
Pasteurella Multocida ◽  
Cell Envelope ◽  
Rabbit Serum ◽  
Antigenic Analysis ◽  
Whole Cells ◽  
Crossed Immunoelectrophoresis ◽  
Serotype 1 ◽  
Determining Factor ◽  
Envelope Antigen

The application of crossed immunoelectrophoresis to the analysis of a reference cytoplasm and cell envelope preparation from Pasteurella multocida serotype 1 revealed antigenic complexity not previously found. At least 55 cytoplasmic and 19 cell envelope antigens were clearly distinguished. Variation of anticytoplasm immunoglobulin concentration was a major determining factor in resolving the maximum array of cytoplasmic antigens.The use of intermediate gel modification of crossed immunoelectrophoresis permitted the recognition of antibodies in the preimmune rabbit serum against a number of cytoplasmic antigens and a single cell envelope antigen. This technique also demonstrated that reference cytoplasm obtained by 105 000 × g centrifugation of sonically disrupted pasteurellae and repeatedly washed reference cell envelope preparation contained antigens of either origin in amounts sufficient to elicit antibody responses in the host. Antisera to whole cells in the intermediate gel indicated that formalin killed P. multocida were capable of eliciting immune responses to both reference systems.

Antigenic analysis of Pasteurella multocida (serotype 1) by crossed immunoelectrophoresis: characterization of whole cell associated antigens

1980 ◽  
Vol 26 (12) ◽  
pp. 1392-1402
Author(s):  
J. L. Bhasin ◽  
L. Lapointe-Shaw
Keyword(s):  
Pasteurella Multocida ◽  
Cell Envelope ◽  
Potassium Thiocyanate ◽  
Antigenic Analysis ◽  
Whole Cell ◽  
Heat Stable ◽  
Crossed Immunoelectrophoresis ◽  
Serotype 1 ◽  
Reference Preparation ◽  
Antigen Antibody

Crossed immunoelectrophoresis and other related quantitative immunoelectrophoretic techniques have been used to elucidate the antigenic complexity of a reference preparation of capsular extract, potassium thiocyanate extract, lipopolysaccharide, heat-stable antigens, and free endotoxin from Pasteurella multocida serotype 1. The reactions of these cellular fractions, in crossed immunoelectrophoresis, with reference anti-whole cell immunoglobulins disclosed five antigens in the capsular extract, seven in the potassium thiocyanate extract, one to three in the lipopolysaccharide, three in the heat-stable antigens, and five in the free endoxin. Comparison of these reference antigen–antibody systems, in crossed immunoelectrophoresis, with intermediate gel containing either a reference anti-cell envelope or anticytoplasmic immunoglobulins not only revealed the presence of additional antigens but also gave insight into the probable cellular origins (i.e., cell surface, cell envelope, or cytoplasm) of various antigens unveiled by reference anti-whole cell immunoglobulins. Using the principle of tandem crossed immunoelectrophoresis and crossed-line immunoelectrophoresis the immunochemical relationships between the antigenic components of these reference antigen–antibody systems were established.

scholarly journals Genetic Characterization of a Cell Envelope-Associated Proteinase from Lactobacillus helveticus CNRZ32

1999 ◽  
Vol 181 (15) ◽  
pp. 4592-4597 ◽  
Author(s):  
Jeffrey A. Pederson ◽  
Gerald J. Mileski ◽  
Bart C. Weimer ◽  
James L. Steele
Keyword(s):  
Cell Surface ◽  
Deletion Mutant ◽  
Cell Envelope ◽  
Physiological Role ◽  
Lactobacillus Helveticus ◽  
Whole Cells ◽  
A Cell ◽  
Hydrolysis Of

ABSTRACT A cell envelope-associated proteinase gene (prtH) was identified in Lactobacillus helveticus CNRZ32. TheprtH gene encodes a protein of 1,849 amino acids and with a predicted molecular mass of 204 kDa. The deduced amino acid sequence of the prtH product has significant identity (45%) to that of the lactococcal PrtP proteinases. Southern blot analysis indicates thatprtH is not broadly distributed within L. helveticus. A prtH deletion mutant of CNRZ32 was constructed to evaluate the physiological role of PrtH. PrtH is not required for rapid growth or fast acid production in milk by CNRZ32. Cell surface proteinase activity and specificity were determined by hydrolysis of αs1-casein fragment 1-23 by whole cells. A comparison of CNRZ32 and its prtH deletion mutant indicates that CNRZ32 has at least two cell surface proteinases that differ in substrate specificity.

scholarly journals Serotyping, Genotyping and Virulence Genes Characterization of Pasteurella multocida and Mannheimia haemolytica Isolates Recovered from Pneumonic Cattle Calves in North Upper Egypt

2020 ◽  
Vol 7 (4) ◽  
pp. 174
Author(s):  
Ahmed H. Abed ◽  
Fawzy R. El-Seedy ◽  
Hany M. Hassan ◽  
Ashraf M. Nabih ◽  
Eman Khalifa ◽  
...  
Keyword(s):  
Pasteurella Multocida ◽  
Confirmatory Test ◽  
Economic Losses ◽  
High Morbidity ◽  
Cattle Industry ◽  
Upper Egypt ◽  
Serotype 1 ◽  
Severe Infections ◽  
Serotype B

Pasteurella (P.) multocida and Mannheimia (M.) haemolytica are the most two common pathogenic bacterial agents causing pneumonia in calves. Both bacteria are associated with significant economic losses in the cattle industry due to high morbidity and mortality rates, especially in the case of severe infections. The objectives of the present study were to perform serotyping and genotyping, as well as characterization of the virulence-associated genes in 48 bacterial isolates; 33 P. multocida and 15 M. haemolytica. All strains were isolated from pneumonic cattle calves showing respiratory manifestations such as fever, nasal discharges, and rapid breathing in North Upper Egypt governorates (Beni-Suef and El-Fayoum). PCR was applied as a confirmatory test using a specific universal gene, kmt1, and rpt2 for P. multocida and M. haemolytica, respectively. The results show that 29 (87.9%) P. multocida and 15 (100%) M. haemolytica isolates were positive for the corresponding universal gene. The results of serotyping indicate that 86.2% of P. multocida isolates belonged to serotype B:2, while 13.8% were untyped. Meanwhile, 60% and 40% of M. haemolytica isolates belonged to serotype 2 and serotype 1, respectively. Investigation of virulence-associated genes showed that all the tested P. multocida isolates harbored nanB, omp87, and toxA genes. Four M. haemolytica isolates harbored both gcp and lktC genes and of these, three isolates harbored the ssa gene. Sequencing of toxA gene of P. multocida and lktC gene of M. haemolytica in the current strains indicated a great homology with strains uploaded in gene banks from different hosts and localities worldwide.

Characterization of an Abnormal Antithrombin (Milano 2) with Defective Thrombin Binding

1986 ◽  
Vol 56 (03) ◽  
pp. 349-352 ◽  
Author(s):  
A Tripodi ◽  
A Krachmalnicoff ◽  
P M Mannucci
Keyword(s):  
Venous Thromboembolism ◽  
Active Site ◽  
Inhibitory Activity ◽  
Antithrombin Iii ◽  
Factor Xa ◽  
Molecular Defect ◽  
Affinity Adsorption ◽  
Italian Family ◽  
Crossed Immunoelectrophoresis

SummaryFour members of an Italian family (two with histories of venous thromboembolism) had a qualitative defect of antithrombin III reflected by normal antigen concentrations and halfnormal antithrombin activity with or without heparin. Anti-factor Xa activities were consistently borderline low (about 70% of normal). For the propositus’ plasma and serum the patterns of antithrombin III in crossed-immunoelectrophoresis with or without heparin were indistinguishable from those of normal plasma or serum. A normal affinity of antithrombin III for heparin was documented by heparin-sepharose chromatography. Affinity adsorption of the propositus’ plasma to human α-thrombin immobilized on sepharose beads revealed defective binding of the anti thrombin III to thrombin-sepharose. Hence the molecular defect of this variant appears to be at the active site responsible for binding and neutralizing thrombin, thus accounting for the low thrombin inhibitory activity.

Characterization of the Serum a1-Antitripsine Level in Primary Spontaneous Pneumotorax Patients

2018 ◽  
Vol 69 (9) ◽  
pp. 2591-2593
Author(s):  
Cristina Grigorescu ◽  
Liviu Ciprian Gavril ◽  
Laura Gavril ◽  
Tiberiu Lunguleac ◽  
Bogdan Mihnea Ciuntu ◽  
...  
Keyword(s):  
Serum Level ◽  
Pulmonary Emphysema ◽  
Spontaneous Pneumothorax ◽  
Antitrypsin Deficiency ◽  
Alpha 1 Antitrypsin Deficiency ◽  
Alpha 1 Antitrypsin ◽  
Determining Factor

Diagnosis of primary or idiopathic spontaneous pneumothorax is one of exclusion, and in fact defines an entity that may have a difficult or impossible cause to be highlighted by current means, we consider it appropriate to study these etiopathogenic aspects. There is a definite association between alpha-1 antitrypsin deficiency and pulmonary emphysema and indirect spontaneous pneumothorax secondary to an emphysematous pulmonary lesion. Dose of alpha-1 antitrypsin is an immunoturbinimetric method for in vitro determination of alpha-1 antitrypsin in human serum and plasma. This product is calibrated to be used for the Daytona RX analyzer. The serum level of alpha-1-antitrypsin is not a determining factor in the postoperative evolution characterized by the interval until air loss disappears, but certainly exerts some influence, the exact level of which remains to be determined.

Immunochemical Characterization of Setaria cervi Microfilarial Antigens Using Novel Antibodies

2019 ◽  
Vol 19 (14) ◽  
pp. 1263-1274 ◽  
Author(s):  
Anuradha Kalani ◽  
Komal Kalani ◽  
Poonam Chaturvedi ◽  
Pankaj Chaturvedi
Keyword(s):  
Adult Worm ◽  
Diagnostic Tests ◽  
Protective Immunity ◽  
Complex Nature ◽  
Immune Recognition ◽  
Setaria Cervi ◽  
Crossed Immunoelectrophoresis ◽  
Study Results ◽  
Immune Rabbit

Background:Filariasis affects millions of people in tropical and subtropical regions of the world and is caused by nematode roundworm. In order to develop a vaccine and specific diagnostic tests, it is important to characterize different stages of the filarial worms. Microfilariae (Mf) stage of the roundworm is found in host’s blood or lymph vessels and can be important not only for developing better immunodiagnostics but also for understanding immune recognition and its relevance to immunepathogenesis and protective immunity.Objective:The present study aimed to immunocharacterize Mf and adult worm antigens that could be helpful in future diagnostic tests.Method:Four different immune sera against Setaria cervi intact live, intact live with adjuvant, intact glutaraldehyde fixed with adjuvant and total somatic Mf were prepared and used for the immunocharacterization of Mf antigens.Results:Our study results suggest that compared to fixed intact Mf, live intact Mf are more immunogenic, as the immune sera generated against intact live Mf showed high ELISA reactivity with Setaria cervi Mf and adult worm antigens. All the four immune sera IgG fractions had surface specificity as determined through considerable ELISA reactivity with S. cervi intact Mf. When tested under native conditions (immunoelectrophoresis and crossed immunoelectrophoresis), all the four immune rabbit sera were able to detect antigens of S. cervi Mf and adult stages.Conclusion:These results can be useful in detailed understanding of the complex nature of the Mf and adult antigens, which are prerequisites in the development of vaccine and more specific diagnostic tests.

scholarly journals Characterization of the SARS-CoV-2 S Protein: Biophysical, Biochemical, Structural, and Antigenic Analysis

ACS Omega ◽  
2020 ◽  
Author(s):  
Natalia G. Herrera ◽  
Nicholas C. Morano ◽  
Alev Celikgil ◽  
George I. Georgiev ◽  
Ryan J. Malonis ◽  
...  
Keyword(s):  
Antigenic Analysis ◽  
S Protein

THE EFFECT OF BACTERIAL CELL LYSIS AND OF PLANT EXTRACTS ON CELLULOSE PRODUCTION BY ACETOBACTER XYLINUM

1963 ◽  
Vol 41 (1) ◽  
pp. 1691-1702 ◽  
Author(s):  
T. E. Webb ◽  
J. Ross Colvin
Keyword(s):  
Bacterial Cell ◽  
Cell Envelope ◽  
Acetobacter Xylinum ◽  
Cellulose Synthesis ◽  
Additive Effects ◽  
Cellulose Production ◽  
Whole Cells ◽  
Negative Results ◽  
Heat Stable ◽  
Plant Sources

The production of cellulose by lysozyme lysates of Acetobacter xylinum is similar to that of a suspension of whole cells, in contrast to the negative results obtained with previous "cell-free" preparations. The results of differential centrifugation of these lysates suggests that most of the enzymes required for cellulose synthesis from glucose normally are held by the cell envelope and are not located in the cytoplasm. However, a heat-stable cofactor(s) is present in the supernatant derived from the cell contents which may stimulate cellulose synthesis by the cell envelopes.The addition of extracts from a number of plant sources increased cellulose synthesis by whole cells of A. xylinum. In particular, the supernatant prepared by centrifugation of an homogenate of tomatoes increased bacterial cellulose production at pH 6 by a factor of 3. Both dialyzable and non-dialyzable substances in the extract are responsible. Fractionation of the non-dialyzable portion of the extract by column chromatography suggests that the overall increase is due to additive effects of several compounds. Here also the compounds appear to act upon the bacterial cell envelope.

Sign in / Sign up

Export Citation Format

Share Document