scholarly journals Production of monoclonal antibodies against Streptococcus mutans antigens

2006 ◽  
Vol 20 (4) ◽  
pp. 297-302 ◽  
Author(s):  
Antonio Carlos Victor Canettieri ◽  
Fujiko Yamasiro Kretchetoff ◽  
Cristiane Yumi Koga-Ito ◽  
Daniella Moreira ◽  
Fabio José Condino Fujarra ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Monoclonal Antibodies ◽  
Streptococcus Mutans ◽  
Cardiac Tissue ◽  
Cross Reactivity ◽  
Oral Streptococci ◽  
Significant Difference ◽  
Common Antigens ◽  
Human Cardiac Tissue

Several studies have been conducted in the last decades aiming to obtain an anti-caries vaccine, however some studies have demonstrated cross reactivity between Streptococcus mutans surface antigens and the human cardiac tissue. In this work, the reactivity of five anti-Streptococcus mutans monoclonal antibodies (MoAb) (24A, 56G, C8, E8 and F6) was tested against oral streptococci, cardiac antigens and skeletal and cardiac myosins, aiming to evaluate the specificity of these MoAb. The hybrid producers of immunoglobulins of the IgG2b class were cloned by limit dilution and expanded in vivo. MoAb were tested by ELISA. The hybrid 24A reacted with S. mutans CCT 1910, S. salivarius CCT 0365 and S. pyogenes T23. No reactivity difference was observed among the tested species. Cross reactivity with heart and cardiac myosin was not confirmed and only reaction with myosin of skeletal muscle was observed (p = 0.0381). The hybrid 56G reacted with all the tested microorganisms and there was statistically significant difference between S. mutans and S. pyogenes T23 (p < 0.001). This hybrid also reacted with myosin of skeletal muscle (p = 0.0095). C8, E8 and F6 presented low reactivity against oral streptococci strains and no reactivity against cardiac antigens. The data of this study showed that the 24A and 56G anti-S. mutans MoAb presented reactivity with S. pyogenes and S. salivarius, reinforcing the occurrence of common antigens between these species. The tested MoAb presented low cross-reactivity with myosin of skeletal muscle, but anti-heart activity could not be confirmed.

Abstract 366: Engineered Human Cardiac Tissue from Skeletal Muscle Derived Cells and Induced Pluripotent Stem Cell Derived Cardiomyocytes

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Kimimasa Tobita ◽  
Jason S Tchao ◽  
Jong Kim ◽  
Bo Lin ◽  
Johnny Huard ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Cardiac Tissue ◽  
Troponin I ◽  
Troponin T ◽  
Induced Pluripotent Stem Cell ◽  
Human Cardiac Tissue ◽  
Induced Pluripotent ◽  
Immature Myocardium

We have previously shown that rat skeletal muscle derived stem cells differentiate into an immature cardiomyocyte (CM) phenotype within a 3-dimensional collagen gel engineered cardiac tissue (ECT). Here, we investigated whether human skeletal muscle derived progenitor cells (skMDCs) can differentiate into a CM phenotype within ECT similar to rat skeletal muscle stem cells and compared the human skMDC-ECT properties with ECT from human induced pluripotent stem cell (iPSc) derived CMs. SkMDCs differentiated into a cardiac muscle phenotype within ECT and exhibited spontaneous beating activity as early as culture day 4 and maintained their activity for more than 2 weeks. SkMDC-ECTs stained positive for cardiac specific troponin-T and troponin-I, and were co-localized with fast skeletal muscle myosin heavy chain (sk-fMHC) with a striated muscle pattern similar to fetal myocardium. The iPS-CM-ECTs maintained spontaneous beating activity for more than 2 weeks from ECT construction. iPS-CM stained positive for both cardiac troponin-T and troponin-I, and were also co-localized with sk-fMHC while the striated expression pattern of sk-fMHC was lost similar to post-natal immature myocardium. Connexin-43 protein was expressed in both engineered tissue types, and the expression pattern was similar to immature myocardium. The skMDC-ECT significantly upregulated expression of cardiac-specific genes compared to conventional 2D culture. SkMDC-ECT displayed cardiac muscle like intracellular calcium ion transients. The contractile force measurements demonstrated functional properties of fetal type myocardium in both ECTs. Our results suggest that engineered human cardiac tissue from skeletal muscle progenitor cells mimics developing fetal myocardium while the engineered cardiac tissue from inducible pluripotent stem cell-derived cardiomyocytes mimics post-natal immature myocardium.

scholarly journals Method for the Selective Measurement of Amino-Terminal Variants of Procalcitonin

2009 ◽  
Vol 55 (9) ◽  
pp. 1672-1679 ◽  
Author(s):  
Joachim Struck ◽  
Martina Strebelow ◽  
Sonja Tietz ◽  
Christine Alonso ◽  
Nils G Morgenthaler ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Bacterial Infections ◽  
Cross Reactivity ◽  
Functional Assay ◽  
Human Endotoxemia ◽  
Amino Terminal ◽  
Endotoxemia Model ◽  
Sandwich Assays ◽  
Kinetics Of

Abstract Background: Procalcitonin (PCT) is an established marker for diagnosing and monitoring bacterial infections. Full-length PCT [116 amino acids that make up procalcitonin (PCT1–116)] can be truncated, leading to des-Ala-Pro-PCT (des-Alanin-Prolin-Procalcitonin; PCT3–116). Current immunoassays for PCT (“total PCT”) use antibodies directed against internal epitopes and are unable to distinguish amino-terminal PCT variants. Here we describe the development of monoclonal antibodies recognizing the amino-termini of PCT1–116 and PCT3–116 and their use in the selective measurement of these PCT species. Methods: With newly developed monoclonal antibodies against the amino-termini of PCT1–116 and PCT3–116, and an antibody against the katacalcin moiety of PCT, we developed and characterized immunoluminometric assays for the 2 PCT peptides. We comparatively assessed the kinetics of PCT variants in a human endotoxemia model. Results: Monoclonal antibodies against the amino-termini of PCT1–116 and PCT3–116 showed &lt;1% cross-reactivity with other PCT-related peptides. The sandwich assays for PCT1–116 and PCT3–116 had functional assay sensitivities of 5 and 1.2 pmol/L, respectively, and exhibited recoveries within 20% of expected values. Plasma PCT1–116 was stable for 6 h at 22 °C and 24 h at 4 °C, and PCT3–116 was stable for at least 24 h at both temperatures. During experimental endotoxemia in healthy people, both PCT1–116 and PCT3–116 increased early in parallel with total PCT, but further increases in PCT1–116 were significantly slower than for PCT3–116 (P = 0.0049) and total PCT (P = 0.0024). Conclusions: The new assays selectively measure PCT1–116 and PCT3–116. Both PCT species increase early during endotoxemia but differ in their kinetics thereafter. The selective measurement of PCT species with different in vivo kinetics may be useful in improving PCT-guided therapies.

scholarly journals Comparative Evaluation of Antibacterial Efficacy of Four Toothpastes and Mouthwashes against Streptococcus mutans and Lactobacillus: An in vivo Study

2016 ◽  
Vol 1 (2) ◽  
pp. 60-65
Author(s):  
Jonathan E Sam ◽  
Paulaian Benin ◽  
Ruth H Beaulah ◽  
Gnanaseelan LNU ◽  
Lal Krishna ◽  
...  
Keyword(s):  
Streptococcus Mutans ◽  
Comparative Evaluation ◽  
Antibacterial Efficacy ◽  
Time Intervals ◽  
Group Iii ◽  
Antimicrobial Substances ◽  
Group I ◽  
Intergroup Comparison ◽  
Significant Difference

ABSTRACT Background Cariogenic microorganisms are the most important cause for occurrence of dental caries. Dentifrices and mouthwashes containing antimicrobial substances are proven to be effective in the eradication of these pathogens from the oral cavity. Aim To evaluate the antimicrobial efficacy of fluoride, chlorhexidine (CHX), herbal, and xylitol containing toothpastes and mouthwashes against Streptococcus mutans (S. mutans) and Lactobacillus (LB) in subjects within the age group of 18 to 22 years at time intervals of 1, 3, and 6 months. Materials and methods One hundred subjects were randomly divided into four groups. Group I: fluoride, group II: chlorhexidine, group III: herbal, group IV: xylitol and instructed to use toothpastes and mouthwashes containing the specific agents. Salivary samples were collected to evaluate the levels of S. mutans and LB at baseline, 1, 3, and 6 months. Bacterial levels were evaluated using caries risk test (CRT) kit. Data were analyzed using analysis of variance and post hoc test. Results During intragroup comparison, S. mutans levels in group I showed statistically significant difference among the four time intervals. On intergroup comparison, S. mutans levels after 6 months for groups I, II, III, and IV were 1.12, 1.16, 1.28, and 1.4 respectively. Conclusion It can be concluded that fluoride, CHX, and xylitol showed a significant reduction in S. mutans and LB count after a time period of 6 months while herbal group did not show a significant reduction in S. mutans and LB count at any intervals. How to cite this article Sam JE, Benin P, Beaulah RH, Gnanaseelan, Krishna L, Raja J. Comparative Evaluation of Antibacterial Efficacy of Four Toothpastes and Mouthwashes against Streptococcus mutans and Lactobacillus: An in vivo Study. J Oper Dent Endod 2016;1(2):60-65.

Steroid effects on permeability of rat thymic lymphocytes to α-aminoisobutyrate

1963 ◽  
Vol 205 (3) ◽  
pp. 446-452 ◽  
Author(s):  
Melvin Blecher
Keyword(s):  
Skeletal Muscle ◽  
Extracellular Fluid ◽  
In Vivo Studies ◽  
Metabolic Inhibitors ◽  
Active Process ◽  
Low Concentrations ◽  
Significant Difference ◽  
Adrenalectomized Rats

In vitro studies of the flux of α-aminoisobutyrate-1-C14 (AIB) between rat thymic lymphocytes and extracellular fluid have revealed that: a) the amino acid enters cells but is not further metabolized; b) at low concentrations, similar to those of amino acids in plasma, the net influx and efflux of AIB exhibit properties of an active process; and c) influx of AIB is inhibited, and efflux stimulated, by deoxycorticosterone (DOC), by metabolic inhibitors, and by other specific steroids. In vivo studies of the distribution of AIB between serum and tissue demonstrated that administration of DOC to adrenalectomized rats inhibited concentration of AIB by thymus, diaphragm, and skeletal muscle, augmented uptake by liver, and increased the serum level of AIB. Prior adrenalectomy of donor rats resulted in no change from normal in the in vitro capacity of thymic lymphocytes to take up AIB. There was no significant difference from normal in the in vivo concentration of AIB by thymus, liver, and skeletal muscle of adrenalectomized rats, although uptake by diaphragm was decreased compared to normal control animals.

31P NMR quantitation of phosphorus metabolites in rat heart and skeletal muscle in vivo

2001 ◽  
Vol 281 (2) ◽  
pp. H882-H887 ◽  
Author(s):  
Sam Hitchins ◽  
Julie M. Cieslar ◽  
Geoffrey P. Dobson
Keyword(s):  
Skeletal Muscle ◽  
Rat Heart ◽  
Signal Amplitude ◽  
Phosphorus Compounds ◽  
External Reference ◽  
Significant Difference ◽  
External Standard ◽  
Phosphorus Metabolites ◽  
Tissue Phosphorus

The aim of this study was to examine two methods of 31P NMR quantitation of phosphocreatine (PCr), ATP, and Pi in rat heart and skeletal muscle in vivo. The first method employed an external standard of phenylphosphonic acid (PPA; 10 mM), and the second method used an enzymatic measurement of tissue ATP equated to the area under the βATP peak. With the use of the external standard, the concentrations of ATP, PCr, and Pi in the rat heart were 4.48 ± 0.33, 9.21 ± 0.65, and 2.25 ± 0.16 μmol/g wet wt, respectively. With the use of the internal ATP standard, measured on the same tissue, the contents (means ± SE) were 4.78 ± 0.19, 9.83 ± 0.18, and 2.51 ± 0.33 μmol/g wet wt, respectively ( n = 7). In skeletal muscle, ATP, PCr, and Pi were 6.09 ± 0.19, 23.44 ± 0.88, and 1.81 ± 0.18 μmol/g wet wt using the PPA standard and 6.03 ± 0.19, 23.30 ± 1.30, and 1.82 ± 0.19 μmol/g wet wt using the internal ATP standard ( n = 6). There was no significant difference for each metabolite as measured by the two methods of quantification in heart or skeletal muscle. The results validate the use of an external reference positioned symmetrically above the coil and imply that each has similar NMR sensitivities (similar signal amplitude per mole of 31P between PPA and tissue phosphorus compounds). We conclude that PCr, ATP, and Pi are nearly 100% visible in the normoxic heart and nonworking skeletal muscle given the errors of measurement.

In vivo insulin signaling through PI3-kinase is impaired in skeletal muscle of adult rat offspring exposed to ethanol in utero

2005 ◽  
Vol 99 (2) ◽  
pp. 528-534 ◽  
Author(s):  
Li Chen ◽  
Xing-Hai Yao ◽  
B. L. G. Nyomba
Keyword(s):  
Insulin Resistance ◽  
Skeletal Muscle ◽  
Insulin Receptor ◽  
Insulin Signaling ◽  
In Utero ◽  
Pi3 Kinase ◽  
Adult Rat ◽  
Rat Offspring ◽  
Significant Difference

It is now known that prenatal ethanol (EtOH) exposure is associated with impaired glucose tolerance and insulin resistance in rat offspring, but the underlying mechanism(s) is not known. To test the hypothesis that in vivo insulin signaling through phosphatidylinositol 3 (PI3)-kinase is reduced in skeletal muscle of adult rat offspring exposed to EtOH in utero, we gave insulin intravenously to these rats and probed steps in the PI3-kinase insulin signaling pathway. After insulin treatment, EtOH-exposed rats had decreased tyrosine phosphorylation of the insulin receptor β-subunit and of insulin receptor substrate-1 (IRS-1), as well as reduced IRS-1-associated PI3-kinase in the gastrocnemius muscle compared with control rats. There was no significant difference in basal or insulin-stimulated Akt activity between EtOH-exposed rats and controls. Insulin-stimulated PKC isoform ζ phosphorylation and membrane association were reduced in EtOH-exposed rats compared with controls. Muscle insulin binding and peptide contents of insulin receptor, IRS-1, p85 subunit of PI3-kinase, Akt/PKB, and atypical PKC isoform ζ were not different between EtOH-exposed rats and controls. Thus insulin resistance in rat offspring exposed to EtOH in utero may be explained, at least in part, by impaired insulin signaling through the PI3-kinase pathway in skeletal muscle.

Profiling of skeletal muscle Ankrd2 protein in human cardiac tissue and neonatal rat cardiomyocytes

2015 ◽  
Vol 143 (6) ◽  
pp. 583-597 ◽  
Author(s):  
Jovana Jasnic-Savovic ◽  
Aleksandra Nestorovic ◽  
Slobodan Savic ◽  
Sinisa Karasek ◽  
Nicola Vitulo ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Cardiac Tissue ◽  
Neonatal Rat ◽  
Rat Cardiomyocytes ◽  
Neonatal Rat Cardiomyocytes ◽  
Human Cardiac Tissue

Sensitive Measurement of Thrombopoietin by a Monoclonal Antibody Based Sandwich Enzyme-linked Immunosorbent Assay

1997 ◽  
Vol 78 (04) ◽  
pp. 1262-1267 ◽  
Author(s):  
Claudia C Folman ◽  
Albert E G K von dem Borne ◽  
Irma H J A M Rensink ◽  
Winald Gerritsen ◽  
C Ellen van der Schoot ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
Platelet Transfusion ◽  
Large Scale ◽  
Limit Of Detection ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Reliable Tool

SummaryIn this report a sensitive enzyme-linked immunosorbent assay (ELISA) for the measurement of plasma thrombopoietin (Tpo) is described that is solely based on monoclonal antibodies (MoAbs).The assay has an intra and inter-assay variance of 5-7% and 7-13%, respectively. Native and recombinant human Tpo (rhTpo) were recognized equally well, no cross reactivity with other cytokines was found and rhTpo added to plasma and serum was completely recovered. With the ELISA, Tpo concentrations in EDTA-anticoagulated plasma of all controls (n = 193) could be determined, since the limit of detection (2 ± 0.8 A.U./ml, mean ± sd) was lower than the concentration found in controls (11 ± 8 A.U./ml, mean ± sd; 2.5th-97.5th percentile: 4-32 A.U./ml). Tpo levels in serum were on average 3.4 times higher than in plasma.We showed in vivo that Tpo is bound by platelets, as in thrombocytopenic patients (n = 5) a platelet transfusion immediately led to a drop in plasma Tpo level, whereas in patients receiving chemotherapy the induced thrombocytopenia was followed by a rise in plasma Tpo levels.In summary, these results indicate that this ELISA is a reliable tool for Tpo measurements and is applicable for large scale studies.

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