QUANTIFICATION OF STREPTOCOCCUS SANGUINIS ISOLATED FROM DENTAL PLAQUE AND SALIVA OF SUBJECTS WITH AND WITHOUT CORONARY HEART DISEASE – ANALYSIS USING REAL-TIME PCR

Coronary Heart Disease (CHD) is the major cause of death in most countries in the world. Gram-positive and Gram-negative bacteria have been identified in bacteremia cases and known to have a role in various vascular diseases, including Streptococcus sanguinis which is most frequently isolated from endocarditis patients and often associated with CHD. The purpose of this study was to analyze the number of Streptococcus sanguinis isolated from dental plaque and saliva of subjects with and without CHD. Bacterial colonies isolated from the dental plaque and saliva of 16 subjects without CHD and 8 subjects with CHD were planted in Mitis salivarius agar, and then the DNA was extracted and quantified with a Real-Time PCR technique using 16S rRNA specific primers. The quantification of Real-Time PCR showed that there was a difference in the number of S. sanguinis between the two groups of subjects, but an unpaired T-test showed that the difference was not statistically significant. Furthermore, the number of S. sanguinis from dental plaque in CHD subjects tends to be higher than that of non-CHD subjects whereas the number of S. sanguinis from saliva in non-CHD subjects tends to be higher than that of CHD subjects.

Download Full-text

Abstract P276: Differential Association of D-dimer with Stroke and Coronary Heart Disease: The REasons for Geographic and Racial Differences in Stroke (REGARDS) Study

Circulation ◽

10.1161/circ.127.suppl_12.ap276 ◽

2013 ◽

Vol 127 (suppl_12) ◽

Author(s):

Neil A Zakai ◽

George Howard ◽

Leslie A McClure ◽

Suzanne E Judd ◽

Brett M Kissela ◽

...

Keyword(s):

Coronary Heart Disease ◽

Heart Disease ◽

Vascular Disease ◽

Racial Differences ◽

Statistical Significance ◽

Female Gender ◽

Cox Models ◽

The Difference ◽

Chd Risk Factors ◽

D Dimer

Introduction: D-dimer, a marker of coagulation activation, has higher levels in blacks than whites and has been variably associated with stroke and coronary heart disease (CHD). Methods: REGARDS recruited 30,239 participants in their homes across the continental US between 2003-07; by design 55% were female, 41% black, and 56% lived in the southeast. In a case-cohort study, D-dimer was measured in 646 participants with incident stroke, 515 with incident CHD, and 1104 in a cohort random sample. D-dimer was log transformed and modeled per 1-unit increase. Cox models were used to determine the HR for vascular disease for D-dimer and the difference in HR (95% CI) by race and vascular disease calculated by bootstrapping with 1000 replicate samples and using the 2.5 and 97.5 percentiles of the distribution (see Table for model variables). Results: Median D-dimer was higher in blacks (0.45 mcg/mL; IQR 0.26, 0.85) than whites (0.38 mcg/mL; IQR 0.23, 0.69); p <0.001. D-dimer was higher with increasing age, female gender, diabetes, hypertension and prebaseline cardiovascular disease (all p <0.05). The table shows the HR of stroke and CHD by baseline D-dimer. In minimally-adjusted models, D-dimer was associated with both stroke and CHD. Accounting for Framingham stroke and CHD risk factors, D-dimer remained associated with CHD (HR 1.45; 95% CI 1.18, 1.79), but was marginally associated with stroke (HR 1.20; 95% CI 0.99, 1.45). The difference in the HR of D-dimer between CHD and stroke was 0.22 in the basic model and 0.25 in the Framingham model, but this difference was of marginal statistical significance (Table). There was no difference in the HRs for stroke or CHD for D-dimer in blacks compared to whites (Table). Discussion: The association of D-dimer with stroke appeared smaller than for CHD with similar associations by race. Findings suggest that hemostasis activation may play a greater role in pathogenesis of CHD than stroke. Further study is needed to confirm these findings and evaluate the association of D-dimer with different stroke subtypes.

Download Full-text

Effect of Tobacco on Lipoprotein Profile: A Comparative study among Smokers and Chewers

Research Journal of Pharmacy and Technology ◽

10.52711/0974-360x.2021.00865 ◽

2021 ◽

pp. 4973-4976

Author(s):

Roshan Kumar Jha ◽

Ranjit S. Ambad ◽

Priya Koundal ◽

Akansha Singh

Keyword(s):

Risk Factors ◽

Coronary Heart Disease ◽

Heart Disease ◽

Total Cholesterol ◽

Sample Processing ◽

Exclusion Criteria ◽

Future Studies ◽

Area Of Interest ◽

Significant Difference ◽

The Difference

It has been proved that tobacco is one of the cholesterol dependent risk factors pathogenically, and in addition with other risk factors it may lead to coronary heart disease. Thus, a strong interaction exists between hypercholesterolemia and tobacco ingesting in the genesis of coronary heart disease. The aim of this study was to study the effect of tobacco smoking and chewing and compare its effect on lipoproteins. 60 subjects were included in the study, and were grouped into 3 three groups, tobacco smokers, tobacco chewers and tobacco non-abusers. Each group comprises 20 participants: selected on the basis of inclusion and exclusion criteria. Proper sampling and sample processing methods were employed to evaluate lipid profile. Total cholesterol and triglycerides levels were increased in smokers in comparison to non-smokers/non-chewers, and the differences were significant p<0.0001. HDL level was decreased in smokers as compared to non-smokers/non-chewers and the difference was statistically significant p<0.0001. Total cholesterol and LDL levels were increased in smokers in comparison to chewers. HDL level was decreased in chewers as compared to chewers. There was no significant association in any of the parameters. Present study observed increased and significant p<0.0001 differences in levels of total cholesterol and triglycerides while, HDL levels were decreased significantly p<0.0001, and also observed there was no significant difference among tobacco smokers and chewers. This may be a new area of interest for future studies.

Download Full-text

Developing, Modifying, and Validating a TaqMan Real-Time PCR Technique for Accurate Identification of Leishmania Parasites Causing Most Leishmaniasis in Iran

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.731595 ◽

2021 ◽

Vol 11 ◽

Author(s):

Reza Fotouhi-Ardakani ◽

Seyedeh Maryam Ghafari ◽

Paul Donald Ready ◽

Parviz Parvizi

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Leishmania Major ◽

Taqman Probe ◽

Amino Acid Permease ◽

Specific Primers ◽

Accurate Identification ◽

Parasitic Load ◽

Species Specific

Many laboratory methods are used to diagnose leishmaniasis because it is characterized by varied symptoms and caused by different Leishmania species. A quantitative real-time PCR method based on a TaqMan probe was developed and modified for accurate identification of human cutaneous leishmaniasis (caused by Leishmania major or Leishmania tropica) from endemic areas of Iran. Two gene regions of amino acid permease 3 (AAP3) and cytochrome oxidase II (COII) were considered. Six new sets of species-specific primers and probes were designed. A total of 123 samples were examined and employed to evaluate and validate real-time PCR. According to parasitic load of the genesig®Leishmania Advanced Standard Kit, a serial dilution of purified plasmid (2–2×107 copies/reaction) was prepared under the same conditions for both genes. Specific primers and probes were able to detect three and six parasite copies in AAP3 and COII genes, respectively, and were able to detect three copies of parasites for L. major and L. tropica. The sensitivities of the reference kit and our method were 98.7 and 98.1%, respectively, and specificity was 100% for detecting parasite genomes in all assays. Designed primers and probes performed well in terms of efficiency and regression coefficient. For AAP3 and COII genes, respectively, the linear log range was 7 and the correlation coefficient (R2) was 0.749 and 0.996 for the reference kit using the standard generated curve and 0.98 and 0.96 with serial dilutions of parasite DNA. This research detected L. major and L. tropica definitely and opens the horizon for the other scientists in the multiplex reactions in designing and optimization of the conditions in silico and in vivo.

Download Full-text

Co-infection of bovine papillomavirus type-1 and -10 in teat warts of dairy cattle

Veterinární Medicína ◽

10.17221/7178-vetmed ◽

2013 ◽

Vol 58 (No. 12) ◽

pp. 605-608

Author(s):

P. Kumar ◽

BL Jangir ◽

G. Saikumar ◽

R. Somvanshi

Keyword(s):

Dairy Cattle ◽

Real Time ◽

Real Time Pcr ◽

Rice Grain ◽

Bovine Papillomavirus ◽

Viral Dna ◽

Specific Primers ◽

Copy Numbers ◽

Pcr Techniques

The present study was carried out to investigate the involvement of different bovine papillomaviruses in the teat warts of cattle. A total of 11 teat wart samples showing rice grain-like and small, sessile elevated greyish or flesh-like growths were collected from dairy cattle. DNA was extracted from these teat wart samples and PCR and real time PCR techniques were applied using specific primers for BPV-1 and -10 to detect the presence of viral nucleic acid. PCR revealed the presence of viral DNA of BPV-1 and -10 in three and seven samples, respectively. Quantification using real time PCR revealed that the copy numbers of the viral DNA of BPV-1 and -10 DNA varied from 1.12E + 04 to 2.99E + 04 and 3.56E + 02 to 5.23E + 06, respectively. From the present study it can be concluded that BPV-1 and -10 are involved in production of rice grain-like and sessile elevated growths on the teats of cattle.

Download Full-text

Non-invasive assessment of endothelial activity in patients with coronary heart disease and cardiovascular risk factors

VASA ◽

10.1024/0301-1526.37.2.137 ◽

2008 ◽

Vol 37 (2) ◽

pp. 137-142 ◽

Cited By ~ 8

Author(s):

Fronek ◽

Allison

Keyword(s):

Risk Factors ◽

Coronary Heart Disease ◽

Heart Disease ◽

Cardiovascular Risk ◽

Cardiovascular Risk Factors ◽

Cvd Risk ◽

Hypertension Status ◽

The Difference ◽

Highly Correlated ◽

And Gender

Background: The aim of this study was first to compare the widely used flow mediated dilation ( FMD ) method with the iontophoretically induced acetylcholine vasodilation (IAV ) procedure. The ultimate goal was to examine the endothelial activity ( EA ) in patients with various cardiovascular risk factors compared with control subjects. Patients and methods: In the upper extremities of 27 subjects, comparisons of EA by FMD and IAV measured with laser Doppler flux method (LDF) were conducted. IAV-EA was then measured using LDF in an additional 93 subjects with various cardiovascular ( CVD ) risk factors and/or a diagnosis of coronary heart disease (CHD). Results: The mean age of the subjects was 56.2 years and 54% were male. There was a robust and significant correlation between FMD vs IAV endothelial activity (r = 0.87, p = 0.025). After adjustment for age, there were significant differences in LDF-measured, acetylcholine-induced EA by diagnosis of CHD (p = 0.02), hyperlipidemia (p = 0.03) and diabetes (p < 0.01), as well as by sex (p < 0.01). The difference by hypertension status was of borderline significance (p = 0.07). LDF EA was higher in non-smokers compared to smokers but this difference was not statistically significant (p = 0.3). After adjustment for age and gender, a 10-unit increase in LDF-measured EA was associated with a 12% lower odds for a diagnosis of CHD (p = 0.07). Conclusions: Measurement of IAV-EA by LDF is a simple, noninvasive methodology which is highly correlated with post-occlusive FMD EA and is also significantly associated with a diagnosis of CHD.

Download Full-text

Real-time PCR Assay with SNP-specific Primers for the Detection of a G143A Mutation Level in Venturia inaequalis Field Populations

Journal of Phytopathology ◽

10.1111/j.1439-0434.2011.01805.x ◽

2011 ◽

Vol 159 (7-8) ◽

pp. 569-578 ◽

Cited By ~ 8

Author(s):

Monika Michalecka ◽

Tadeusz Malinowski ◽

Agata Broniarek-Niemiec ◽

Anna Bielenin

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Venturia Inaequalis ◽

Pcr Assay ◽

Specific Primers

Download Full-text

Quantitative PCR with 16S rRNA-Gene-Targeted Species-Specific Primers for Analysis of Human Intestinal Bifidobacteria

Applied and Environmental Microbiology ◽

10.1128/aem.70.1.167-173.2004 ◽

2004 ◽

Vol 70 (1) ◽

pp. 167-173 ◽

Cited By ~ 309

Author(s):

Takahiro Matsuki ◽

Koichi Watanabe ◽

Junji Fujimoto ◽

Yukiko Kado ◽

Toshihiko Takada ◽

...

Keyword(s):

Real Time ◽

Quantitative Pcr ◽

Real Time Pcr ◽

Intestinal Flora ◽

Healthy Adults ◽

Pcr Detection ◽

Distribution Analysis ◽

Specific Primers ◽

Cell Counts ◽

Species Specific

ABSTRACT A highly sensitive quantitative PCR detection method has been developed and applied to the distribution analysis of human intestinal bifidobacteria by combining real-time PCR with Bifidobacterium genus- and species-specific primers. Real-time PCR detection of serially diluted DNA extracted from cultured bifidobacteria was linear for cell counts ranging from 106 to 10 cells per PCR assay. It was also found that the method was applicable to the detection of Bifidobacterium in feces when it was present at concentrations of >106 cells per g of feces. Concerning the distribution of Bifidobacterium species in intestinal flora, the Bifidobacterium adolescentis group, the Bifidobacterium catenulatum group, and Bifidobacterium longum were found to be the three predominant species by examination of DNA extracted from the feces of 46 healthy adults. We also examined changes in the population and composition of Bifidobacterium species in human intestinal flora of six healthy adults over an 8-month period. The results showed that the composition of bifidobacterial flora was basically stable throughout the test period.

Download Full-text

Accounting for Time-Varying Confounding in the Relationship Between Obesity and Coronary Heart Disease: Analysis With G-Estimation

American Journal of Epidemiology ◽

10.1093/aje/kwx360 ◽

2017 ◽

Vol 187 (6) ◽

pp. 1319-1326 ◽

Cited By ~ 13

Author(s):

Maryam Shakiba ◽

Mohammad Ali Mansournia ◽

Arsalan Salari ◽

Hamid Soori ◽

Nasrin Mansournia ◽

...

Keyword(s):

Coronary Heart Disease ◽

Heart Disease ◽

Time Varying ◽

The Relationship ◽

Disease Analysis

Download Full-text

Differentiation of infectious bronchitis virus vaccine strains Ma5 and 4/91 by TaqMan real-time PCR

Polish Journal of Veterinary Sciences ◽

10.1515/pjvs-2017-0073 ◽

2017 ◽

Vol 20 (3) ◽

pp. 599-601 ◽

Cited By ~ 5

Author(s):

T. Stenzel ◽

D. Dziewulska ◽

M. Śmiałek ◽

B. Tykałowski ◽

J. Kowalczyk ◽

...

Keyword(s):

Real Time ◽

Virus Vaccine ◽

Infectious Bronchitis Virus ◽

Real Time Pcr ◽

Cross Reactivity ◽

Specific Primers ◽

Assay Sensitivity ◽

Infectious Bronchitis ◽

Molecular Assays ◽

Rapid Molecular Assays

Abstract The aim of this study was to develop rapid molecular assays for differentiating vaccine strains Ma5 and 4/91 of the infectious bronchitis virus (IBV). Specific primers and probes for S1 and N genes were designed based on the nucleotide sequences of both vaccine strains. Cross-reactivity was not observed. Assay sensitivity was 2.373 × 103 copies of the Ma5 strain, and 3.852 x 103 copies of the 4/91 strain. Samples belonging to a known genotype demonstrated that the designed assays supported rapid and sensitive detection of Ma5 and 4/91 vaccine strains of IBV.

Download Full-text