Investigating the Impact of Serum on the Growth of TM7x and Host XH001

2020 ◽  
Vol 9 (2) ◽  
Author(s):  
Alice Khomski ◽  
Megan Pugach-Gordon
Keyword(s):  
Periodontal Disease ◽  
Culture Medium ◽  
Considerable Increase ◽  
Gingival Crevicular Fluid ◽  
Fetal Bovine Serum ◽  
Colony Forming Units ◽  
Fetal Bovine ◽  
Crevicular Fluid ◽  
Potential Pathogenicity ◽  
The Impact

Nanosynbacter lyticus strain TM7x is an ultra-small, parasitic bacterium that lives on the surface of host, Actinomyces odontolyticus strain XH001. Studies have reported a considerable increase in TM7x abundance in patients with periodontal disease, and this association warrants further investigation. Interestingly, gingival crevicular fluid (GCF) flow also increases during periodontitis. The prevalence of TM7x during periodontal disease may thus result from attraction to GCF. However, GCF is not commercially available for testing. Since its composition is similar to that of fetal bovine serum (FBS), FBS was tested instead. The objectives of this project are to determine whether FBS promotes the growth of TM7x and XH001 and to develop the optimal culture medium for analysis of these microbes. TM7x/XH001 co-culture and XH001 monoculture were separately cultivated in media (RPMI and FMC) supplemented with FBS. Cultures grown in the same, non-supplemented media served as control samples. Serial dilutions were performed and spotted onto plates at several time points, and samples were quantified in colony-forming units (CFU)/mL. 2% FBS FMC induced a significant increase in the growth of culture samples, as compared to control FMC (p = 0.0140 and p = 0.0424 for the co-culture and monoculture, respectively). Furthermore, microscopy revealed a heightened presence of TM7x in FBS-supplemented media in comparison to control media. These findings signify that FBS is an effective supplement to RPMI and FMC, both of which support TM7x/XH001 growth. Overall, the utilization of these media with FBS can ensure optimal cultivation of TM7x for investigation of its potential pathogenicity.

Involvement of nucleus in the maturation of dengue-2 virus in C6/36 cells

1990 ◽  
Vol 48 (3) ◽  
pp. 912-913
Author(s):  
Jaang J. Wang ◽  
Cheng C. Chen ◽  
Men F. Shaio ◽  
Chia T. Liu ◽  
Chung S. Lee ◽  
...  
Keyword(s):  
Culture Medium ◽  
Cytopathic Effect ◽  
Fetal Bovine Serum ◽  
Virus Particles ◽  
Electron Microscopies ◽  
Embedding Technique ◽  
Fetal Bovine ◽  
Flat Embedding ◽  
20 Nm ◽  
Labeling Method

The involvement of nucleus in the maturation processes of Dengue-2 virus in a mosquito cell line, C6/36 cells, has been identified by the electron microscopy and immunocytochemistry. The C6/36 cells were obtained from ATCC and maintained in MEM culture medium containing 10% fetal bovine serum at 28°C. The cell suspensions or cells grown on teflon-coated coverslips were infected with Dengue-2 virus (107/ml) for various time periods of 2 hours, 3, 6, 8, and 10 days. The cells were then fixed in buffered 1.5% glutaraldehyde, and washed in acetone before immunolabeled with monoclonal antibody. An indirect immunocytochemical labeling method of avidin-biotin complex (ABC) conjugated with peroxidase or gold particles (20 nm in diameter) and a flat embedding technique were used to localize the virus particles.At early stages of infections (before 3 days), there were no virion particles detected. After 6 days and on of infections, cytopathic effect (CPE) was observed and showed positive immuno-peroxidase reactions under the light and electron microscopies.

Salivary Diagnosis - Clinical Uses in Assessing Oral Inflammation

2017 ◽  
Vol 68 (6) ◽  
pp. 1201-1204 ◽  
Author(s):  
Iulia Ioana Stanescu ◽  
Alexandra Totan ◽  
Florentina Rus ◽  
Daniela Miricescu ◽  
Brandusa Mocanu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Periodontal Disease ◽  
Gingival Crevicular Fluid ◽  
Oxidative Stress Marker ◽  
Clinical Settings ◽  
Tissue Destruction ◽  
Whole Saliva ◽  
Oral Inflammation ◽  
Positive Correlations ◽  
Crevicular Fluid

The past decades demonstrated that saliva and its components represent a remarkable diagnosis fluid with valuable clinical uses for both oral and systemic diseases. At the same time it is well established that oxidative stress is involved in a wide number of pathologies, including periodontitis. The specific aim of the present study which included 50 subjects is to determine if saliva can be used in clinical settings to correlate oxidative stress and tissue destruction markers with the severity of periodontal disease. An important oxidative stress marker - 8-hydroxydesoxyguanosine (8-OHdG) and a collagen degradation marker - beta-crosslaps (b-CTX) were quantified in both saliva and gingival crevicular fluid (GCF) using ELISA kits and were found to be significantly increased in the chronic periodontitis group when compared to respective controls (p[0.05). At the same time positive correlations were observed between whole saliva and gingival crevicular fluid (p[0.05). Significant correlations were also determined between GCF and salivary markers and clinical parameters of periodontal disease. Present results demonstrate that saliva and its components can successfully be used in clinical settings and represents a reliable tool for assessing periodontal disease severity.

scholarly journals NLRP3 Inflammasome Expression in Gingival Crevicular Fluid of Patients with Periodontitis and Chronic Hepatitis C

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Petra Surlin ◽  
Luminita Lazar ◽  
Cerasella Sincar ◽  
Dorin Nicolae Gheorghe ◽  
Dora Maria Popescu ◽  
...  
Keyword(s):  
Chronic Hepatitis ◽  
Hepatitis C ◽  
Periodontal Disease ◽  
Chronic Hepatitis C ◽  
Inflammatory Reaction ◽  
Nlrp3 Inflammasome ◽  
Gingival Crevicular Fluid ◽  
Periodontal Status ◽  
Local Inflammatory Reaction ◽  
Crevicular Fluid

The study is aimed at assessing the impact that periodontal disease and chronic hepatitis C could have on gingival crevicular fluid levels of the NLRP3 inflammasome, caspase-1 (CASP-1), and interleukin-18 (IL-18) and at evaluating whether the increased local inflammatory reaction with clinical periodontal consequences is correlated to their upregulation. Patients were divided into four groups, according to their periodontal status and previously diagnosed hepatitis C, as follows: (i) CHC group, chronic hepatitis C patients; (ii) P group, periodontal disease patients, systemically healthy; (iii) CHC + P group, patients suffering from both conditions; and (iv) H group, systemically and periodontally healthy controls. Gingival crevicular samples were collected for quantitative analysis of the NLRP3 inflammasome, CASP-1, and IL-18. CHC + P patients expressed the worse periodontal status and the highest NLRP3, CASP-1, and IL-18 levels, the difference being statistically significant ( p < 0.05 ). The P group patients also expressed significantly more elevated NLRP3, CASP-1, and IL-18 levels, as compared to nonperiodontal patients (CHC and H groups). Chronic hepatitis C and periodontal disease could have a significant influence on the upregulation of NLRP3 inflammasome and its components, possibly contributing to an increased local inflammatory reaction and clinical periodontal consequences.

Fetal bovine serum induces changes in fatty acid composition ofTrypanosoma cruziphosphoinositides

1995 ◽  
Vol 41 (10) ◽  
pp. 951-954 ◽  
Author(s):  
G. Racagni ◽  
M. G. de Lema ◽  
G. Hernández ◽  
E. E. Machado-Domenech
Keyword(s):  
Fatty Acids ◽  
Fatty Acid Composition ◽  
Fatty Acid ◽  
Trypanosoma Cruzi ◽  
Acid Composition ◽  
Bovine Serum ◽  
Culture Medium ◽  
Culture Media ◽  
Fetal Bovine Serum ◽  
Fetal Bovine

Fetal bovine serum (FBS) is a necessary constituent of the culture media employed to foster the growth of Trypanosoma cruzi epimastigote forms. In different laboratories, the serum is used at final concentrations of 5 or 10%. We have normally supplemented the complex medium with 10% FBS. Under this condition we have described the fatty acid composition of the total lipids and of the phosphoinositide fractions. Additionally, we have reported the increase of polyphosphoinositides and phosphatidic acid after cholinergic stimulation. Since further attempts to reproduce these results with 5% FBS in the culture medium were not successful, the effect of the FBS concentration on the fatty acid composition of phospholipids from the T. cruzi epimastigote forms was thoroughly examined. This work showed that when the FBS concentration supplementing the culture medium was reduced from 10 to 5%, the fatty acid composition of the phosphoinositides was altered while the other major phospholipids were not significantly affected. The most relevant result was the decrease in the content of linoleic acid (18:2) and the increase of palmitoleic acid (16:1) in phosphatidylinositol 4,5-bisphosphate. Phosphatidylinositol (PI) and phosphatidylinositol phosphate also exhibited similar changes in the same fatty acids. The C2fatty acid composition of the phosphoinositides, under the same conditions, is also reported here for the first time.Key words: Trypanosoma cruzi, fatty acids, phosphoinositides, fetal bovine serum, phospholipids.

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