scholarly journals ABO Genotyping from Pulp and Dentin Using DNA - A PCR Study

Author(s):  
Jaya Lakshmi B. ◽  
Avinash Tejasvi M. L.

Background: Blood grouping has a major role in forensic science in the field of human identification. Aim of the Study: Aim of this study was to determine ABO genotyping from pulp and dentin using PCR method. Objectives: To determine PCR based blood grouping from the DNA isolated from tooth Pulp and Dentin. Materials and Meathods: Dental pulp tissue and Dentin was collected from 20 permanent teeth and DNA extraction was carried out from pulp using (modified proteinase –K method) and from dentin using phenol/chloroform(organic method). PCR procedure was carried out and samples were subjected to agarose gel electrophoresis and blood group was determined using specific PCR primers for ABO genotyping. Results: Among the 20 samples of pulp tissue, 17 samples showed accurate results in PCR method in comparison with slide agglutination method. Among the 20 samples of dentin, blood grouping from dentin was not possible as the quantity and purity of DNA from dentin samples were not optimal. Sensitivity of 85% and specificity of 50% was noticed from the samples of pulp. Conclusion: PCR was found to be an effective method in determination of blood group from teeth. Thus our study states that isolation of DNA can be done from both pulp and dentin and the blood grouping can be done from tooth pulp by PCR method. Hence PCR method can be used for identification of individuals which adds beneficiary value to forensic dentistry.

2003 ◽  
Vol 69 (12) ◽  
pp. 7430-7434 ◽  
Author(s):  
Trevor G. Phister ◽  
David A. Mills

ABSTRACT Traditional methods to detect the spoilage yeast Dekkera bruxellensis from wine involve lengthy enrichments. To overcome this difficulty, we developed a quantitative real-time PCR method to directly detect and enumerate D. bruxellensis in wine. Specific PCR primers to D. bruxellensis were designed to the 26S rRNA gene, and nontarget yeast and bacteria common to the winery environment were not amplified. The assay was linear over a range of cell concentrations (6 log units) and could detect as little as 1 cell per ml in wine. The addition of large amounts of nontarget yeasts did not impact the efficiency of the assay. This method will be helpful to identify possible routes of D. bruxellensis infection in winery environments. Moreover, the time involved in performing the assay (3 h) should enable winemakers to more quickly make wine processing decisions in order to reduce the threat of spoilage by D. bruxellensis.


2006 ◽  
Vol 30 (1) ◽  
pp. 19-21 ◽  
Author(s):  
Manju Gopa Kumar ◽  
Amitha Hegde

Teeth endure postmortem degradation and extreme changes in ambient temperature and pressure better than most human tissues. In the present day scenario the growing number of crime against children in the form of battering, physical/sexual abuse, abduction and kidnapping, the use of exfoliated primary teeth, become many times the only evidence available at the crime scene. Despite exposure of the body to burial, mutilation, explosion or incineration, it is usually possible to extract DNA from pulp tissue of tooth with sufficient quality and quantity. Hence the present study was undertaken to find out the sex of a child from exfoliated/extracted deciduous teeth using a Polymerase Chain reaction(PCR) based analysis. Tooth samples were stored in room temperature after double coding for various periods. Dental pulp tissue was collected from each sample and DNA was isolated by proteinase-k digestion and phenol chloroform extraction methods. PCR amplification was done with two sets of oligonucleiotide primers. Amplification of X (131bp) and Y-specific sequences (172bp) in males and that of the X-specific sequence in females was observed and compared with the template DNA showing male and female controls. Determination of sexes of all freshly collected samples within 24 hours and after 1 month of extraction respectively gave 100% result. However, PCR was not found to be an effective method for sex determination after 6 months post extraction.


1993 ◽  
Vol 295 (1) ◽  
pp. 313-315 ◽  
Author(s):  
A A Fryer ◽  
L Zhao ◽  
J Alldersea ◽  
W R Pearson ◽  
R C Strange

We describe the identification of the GSTM1 null, GSTM1 A, GSTM1 B and GSTM1 A,B polymorphisms at the glutathione S-transferase GSTM1 locus using a single-step PCR method. Target DNA was amplified using primers to intron 6 and exon 7 with site-directed mutagenesis being used to introduce a restriction site in DNA amplified from GSTM1 *A, thereby allowing differentiation of this allele and GSTM1 *B. The accuracy of this approach in identifying the GSTM1 A, GSTM1 B, GSTM1 A,B and GSTM1 null polymorphisms was confirmed by comparison with, firstly, an established PCR method that distinguishes GSTM1 *0 homozygotes from individuals with the other GSTM1 genotypes and, secondly, GSTM1 phenotypes determined using chromatofocusing.


2019 ◽  
Vol 47 (10) ◽  
pp. 5094-5105
Author(s):  
Yan Huang ◽  
Xiaoying Tang ◽  
Zafer C. Cehreli ◽  
Xiaoyun Dai ◽  
Jiangjingjun Xu ◽  
...  

Objectives To investigate the potential for pulpal regeneration via autologous transplantation of deciduous tooth pulp into immature necrotic permanent teeth using an experimental dog model. Methods Experimental apical periodontitis was induced in 60 teeth of six Beagle dogs. Following canal disinfection and pulpotomy, autologous deciduous pulp tissue was transplanted into the root canals (n = 30); as controls, contralateral teeth were treated in accordance with the recommendations of the American Association of Endodontists. Radiographic examinations were performed immediately before transplant, as well as 3 and 6 months after transplant. At the 6-month examination, root samples were collected and histological and immunohistochemical analyses were used to examine tissue regeneration. Results Radiographic analysis showed no significant differences in most histopathological parameters examined; however, apical diameter reduction was greater in the experimental group. Histological and immunohistochemical analyses showed that the canal walls of the experimental group had newly formed dentin-like tissue with dentinal tubules, while the control group had cementum-like deposits along the canal wall and apical foramina. Conclusions Autologous transplantation may be useful for regeneration of dental pulp in necrotic young permanent teeth.


Author(s):  
Minu Anoop ◽  
Indrani Datta

: Most conventional treatments for neurodegenerative diseases fail due to their focus on neuroprotection rather than neurorestoration. Stem cell‐based therapies are becoming a potential treatment option for neurodegenerative diseases as they can home in, engraft, differentiate and produce factors for CNS recovery. Stem cells derived from human dental pulp tissue differ from other sources of mesenchymal stem cells due to their embryonic neural crest origin and neurotrophic property. These include both dental pulp stem cells [DPSCs] from dental pulp tissues of human permanent teeth and stem cells from human exfoliated deciduous teeth [SHED]. SHED offer many advantages over other types of MSCs such as good proliferative potential, minimal invasive procurement, neuronal differentiation and neurotrophic capacity, and negligible ethical concerns. The therapeutic potential of SHED is attributed to the paracrine action of extracellularly released secreted factors, specifically the secretome, of which exosomes is a key component. SHED and its conditioned media can be effective in neurodegeneration through multiple mechanisms, including cell replacement, paracrine effects, angiogenesis, synaptogenesis, immunomodulation, and apoptosis inhibition, and SHED exosomes offer an ideal refined bed-to-bench formulation in neurodegenerative disorders. However, in spite of these advantages, there are still some limitations of SHED exosome therapy, such as the effectiveness of long-term storage of SHED and their exosomes, the development of a robust GMP-grade manufacturing protocol, optimization of the route of administration, and evaluation of the efficacy and safety in humans. In this review, we have addressed the isolation, collection and properties of SHED along with its therapeutic potential on in vitro and in vivo neuronal disorder models as evident from the published literature.


Materials ◽  
2021 ◽  
Vol 14 (2) ◽  
pp. 305
Author(s):  
Chung-Min Kang ◽  
Saemi Seong ◽  
Je Seon Song ◽  
Yooseok Shin

The use of hydraulic silicate cements (HSCs) for vital pulp therapy has been found to release calcium and hydroxyl ions promoting pulp tissue healing and mineralized tissue formation. The present study investigated whether HSCs such as mineral trioxide aggregate (MTA) affect their biological and antimicrobial properties when used as long-term pulp protection materials. The effect of variables on treatment outcomes of three HSCs (ProRoot MTA, OrthoMTA, and RetroMTA) was evaluated clinically and radiographically over a 48–78 month follow-up period. Survival analysis was performed using Kaplan–Meier survival curves. Fisher’s exact test and Cox regression analysis were used to determine hazard ratios of clinical variables. The overall success rate of MTA partial pulpotomy was 89.3%; Cumulative success rates of the three HSCs were not statistically different when analyzed by Cox proportional hazard regression analysis. None of the investigated clinical variables affected success rates significantly. These HSCs showed favorable biocompatibility and antimicrobial properties in partial pulpotomy of permanent teeth in long-term follow-up, with no statistical differences between clinical factors.


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