Antimicrobial Efficacy of Intracanal Medicaments against E. Faecalis Bacteria in Infected Primary Molars by Using Real-Time PCR: A Randomized Clinical Trial

Aim. This study aimed to compare the antimicrobial efficacy of calcium hydroxide (CH) and triple antibiotic paste (TAP) against E. faecalis bacteria in infected primary molars. Methods and Materials. Thirty-nine 4–6-year-old children with an infected primary molar were randomly divided into three equal groups (n = 13) to receive either CH or TAP and an untreated control group. Following access cavity preparation, the first microbiological samples (S1) were collected by using absorbent paper points. The canals were prepared and rinsed. Then, CH or TAP was applied in the root canals. Seven days later, the second microbiological samples (S2) were collected. DNA extraction was performed to count E. faecalis bacteria by using real-time PCR for S1 and S2 samples. Data were analyzed through one-way ANOVA and Tukey’s test (α = 0.05). Results. E. faecalis bacteria counts decreased significantly in CH and TAP groups compared with the control group ( P ≤ 0.001 ). However, no statistically significant difference existed between these two groups ( P = 0.698 ). Conclusion. Both TAP and CH have significant antimicrobial effects as intracanal medicament between the treatment sessions in infected primary teeth.

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Success Rate of Obturation of Root Canals by Different Techniques in Primary Molars: A Comparative Clinical Study

Current Dentistry ◽

10.2174/2542579x02999200503032916 ◽

2020 ◽

Vol 2 (1) ◽

pp. 36-44

Author(s):

Satyawan G. Damle ◽

Ritika Bansal ◽

Dhanashree D. Sakhare

Keyword(s):

Success Rate ◽

Study Groups ◽

Primary Molars ◽

Control Group ◽

Second Primary ◽

Chi Square ◽

Exact Test ◽

Root Canals ◽

Significant Difference ◽

Comparative Clinical Study

Objective: To compare the success rate of different obturation procedures in primary mandibular second molars clinically and also by digital radiovisiography. Methods: A total of 40 children aged between 4-8 years with deeply carious mandibular second primary molars indicated for single session pulpectomy were selected. Canals were obturated with Metapex. The 3 study groups (Endodontic plugger, Handheld lentulospiral, Navi Tip syringe) were compared with the control group (reamer) both clinically and radiovisiographically. The data collected were statistically analyzed using Pearson’s Chi-square and Fisher’s exact test. Results: The use of Navi tip syringe led to the least number of voids followed by Endodontic plugger and Reamer and the highest number of voids was reported with Lentulospiral. Navitip presented maximum number of optimally filled cases followed by Endodontic plugger and Lentulospiral and least number of optimally filled cases with reamer. However, there was no statistically significant difference (p>0.05) in any of the groups with clinical (pain and tenderness to percussion) and radiographic parameters (presence or absence of voids and length of obturation). Conclusion: Within the limitations of the present study, though the clinical outcome was statistically insignificant, Navitip syringe exhibited encouraging results and is a promising option for obturation in primary teeth.

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The association of male infertility with telomere length: A case control study

Indian Journal of Clinical Anatomy and Physiology ◽

10.18231/j.ijcap.2021.070 ◽

2021 ◽

Vol 8 (4) ◽

pp. 325-332

Author(s):

Kate Deepali Rajesh ◽

Puranam Vatsalaswamy ◽

Manvikar Purshotam Rao

Keyword(s):

Male Infertility ◽

Telomere Length ◽

Real Time ◽

Real Time Pcr ◽

Case Control Study ◽

Case Control ◽

Control Group ◽

Significant Difference ◽

Control Study ◽

Sperm Telomere Length

To study the relevance of sperm telomere length and infertility in men. : Our case-control study included twenty-five males in couple with sub-fertility/infertility (test group) and twenty five healthy males (control group) with proven paternity in the age group 25 to 35 years. The Absolute Sperm Telomere length (aSTL) was measured by real-time PCR. We investigated whether any significant difference in the aSTL value existed between the groups and analysed the relationship between aSTL and other sperm parameters.The mean (SE) aSTL recorded in the infertile cases was significantly shorter than for the control group being 140.60 (6.66) Kb/genome and 239.63 (12.32) Kb/genome respectively (p <0.001) A weak correlation was eminent between aSTL kb/genome and the total sperm count mil/ml (rho= 0.04, p - 0.86), progressive sperm motility (rho= - 0.02, p=0.934) and sperm viability (rho= - 0.07 p=0.741) in the infertile group. The measurement of aSTL by real-time PCR is a simple and rapid method that offers further paramount information with respective to the quality of sperm. It is befitted for epidemiological studies, hence opening new perspectives in the evaluation of male infertility. Limitations - Our study was confined to men aged between 25 and 35 years. Further comparative studies are needed to explore the significance of STL and infertility in older males. Additional studies will help illumine the significance of aSTL as a prognostic biomarker in assisted reproduction.

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An in Vivo Comparison of Antimicrobial Efficacy of Sodium Hypochlorite and Biopure MTAD™ against Enterococcus Faecalis in Primary Teeth: A qPCR Study

Journal of Clinical Pediatric Dentistry ◽

10.17796/jcpd.39.1.c4q2155r16817219 ◽

2014 ◽

Vol 39 (1) ◽

pp. 30-34 ◽

Cited By ~ 3

Author(s):

SG Tulsani ◽

N Chikkanarasaiah ◽

S Bethur

Keyword(s):

Root Canal ◽

Primary Teeth ◽

Antimicrobial Efficacy ◽

Permanent Teeth ◽

Control Group ◽

Statistical Significant Difference ◽

Anterior Teeth ◽

Significant Difference ◽

Root Canal Irrigant

Objectives: Biopure MTAD™, a new root canal irrigant has shown promising results against the most common resistant microorganism, E. faecalis, in permanent teeth. However, there is lack of studies comparing its antimicrobial effectiveness with NaOCl in primary teeth. The purpose of this study was to compare the in vivo antimicrobial efficacy of NaOCl 2.5% and Biopure MTAD™ against E. faecalis in primary teeth. Study design: Forty non vital single rooted primary maxillary anterior teeth of children aged 4-8 years, were irrigated either with NaOCl 2.5% (n=15), Biopure MTAD™ (n=15) and 0.9% Saline (n=10, control group). Paper point samples were collected at baseline (S1) and after chemomechanical preparation (S2) during the pulpectomy procedure. The presence of E. faecalis in S1 & S2 was evaluated using Real time Polymerase Chain Reaction. Results: Statistical significant difference was found in the antimicrobial efficacy of NaOCl 2.5 % and BioPure MTAD™ when compared to saline (p>0.05). However, no statistical significant difference was found between the efficacies of both the irrigants. Conclusions: NaOCl 2.5% and BioPure MTAD™, both irrigants are equally efficient against E. faecalis in necrotic primary anterior teeth. MTAD is a promising irrigant, however clinical studies are required to establish it as ideal root canal irrigant in clinical practice.

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Genome-wide screening of m6A-modified transcript in the Wilms’ tumor

10.21203/rs.3.rs-197376/v1 ◽

2021 ◽

Author(s):

Zhuo Liu ◽

Feng He ◽

Jing Liu ◽

Shengrong OuYang ◽

Zexi Li ◽

...

Keyword(s):

Gene Ontology ◽

Real Time ◽

Real Time Pcr ◽

Wilms Tumor ◽

Expression Patterns ◽

Mrna Levels ◽

Gene Ontology Analysis ◽

Quantitative Real Time Pcr ◽

Related Factors ◽

Significant Difference

Abstract Background Wilms’ tumor, also called nephroblastoma, is the most common pediatric renal malignancy. The pathogenesis of Wilms’ tumor has been attributed to several genetic and epigenetic factors. However, the most pervasive internal mRNA modification that affects almost every process of RNA metabolism, RNA N6-Methyladenosine (m6A) methylation, has not been characterized in Wilms’ tumor. Methods Wilms’ tumor (WT) and adjacent non-cancerous (NC) tissue samples were obtained from 23 children with nephroblastoma, and the global m6A levels were measured by mass spectrometry. Analyses by m6A-mRNA epitranscriptomic microarray and mRNA microarray were performed, and m6A-related mRNAs were validated by quantitative real-time PCR for input and m6A-immunoprecipitated RNA samples from WT and NC tissues. Gene ontology analysis and KEGG pathway analysis were performed for differentially expressed genes, and expression of RNA methylation-related factors was measured by quantitative real-time PCR. Results The total m6A methylation levels in total RNA of WT samples and NC samples were (0.21 ± 0.01)% and (0.22 ± 0.01)%, respectively, with no statistically significant difference. Fifty-nine transcripts were differentially m6A-methylated between the WT and NC groups, which showed distinct m6A modification patterns. Gene ontology analysis indicated that m6A-modified genes were enriched in cancer-associated pathways, including the mTOR pathway, and conjoint analysis of the unique methylation and gene expression patterns in WT samples suggested an association with metabolic pathways.The mRNA levels of the m6A-related “reader” genes, YTHDF1, YTHDF2 and IGF2BP3, were statistically higher in WT samples than in NC samples. Conclusion This is the first study to determine the m6A modification profiles in Wilms’ tumor. Our data provide novel information regarding patterns of m6A modification that correlate with carcinogenesis in Wilms’ tumor.

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Resorbable Collagen Barrier Impeding the Extrusion of Obturating Material in Primary Molars Undergoing Resorption – A Randomized Clinical Trial

Journal of Clinical Pediatric Dentistry ◽

10.17796/1053-4625-45.5.4 ◽

2021 ◽

Vol 45 (5) ◽

pp. 312-316

Author(s):

Mishra Neha Sanjeev ◽

Harsimran Kaur ◽

Sandeep Singh Mayall ◽

Rishika ◽

Ramakrishna Yeluri

Keyword(s):

Test Group ◽

Primary Molars ◽

Control Group ◽

Chi Square ◽

Significance Level ◽

Significant Difference ◽

Chi Square Test ◽

Group A ◽

Group B ◽

And Control

Objective: To evaluate the effectiveness of placing a resorbable collagen barrier in impeding the extrusion of obturation material in primary molars undergoing resorption. Study design: All the 94 canals in 47 mandibular molars were allocated to 2 groups- Group ‘A’- 47 canals with collagen barrier (Test group) and Group ‘B’- 47 canals without collagen barrier (Control group) based on randomization protocol. Pulpectomy was performed and obturation of both test and control canals were radiographically assessed. Pearson’s chi – square test was applied to analyze the results. The significance level was predetermined at p < 0.05. Results: Among the test group, 93.6% of the canals showed no extrusion while, 6.4% showed visible extrusion of the material outside the apex. In the control group, 83% showed no extrusion whereas 17% of the canals showed visible extrusion outside the apex. But no significant difference was noted (p>0.05). Conclusion: The placement of resorbable collagen barrier in the apical third of the canal prevented the extrusion of obturating material beyond the apex in resorbing primary molars.

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Evaluation of the Sealing Ability of Three Obturation Techniques Using a Glucose Leakage Test

BioMed Research International ◽

10.1155/2017/2704094 ◽

2017 ◽

Vol 2017 ◽

pp. 1-8 ◽

Cited By ~ 2

Author(s):

Katarzyna Olczak ◽

Halina Pawlicka

Keyword(s):

Continuous Wave ◽

Negative Control Group ◽

Positive Control ◽

Negative Control ◽

Control Group ◽

Thermal Methods ◽

Root Canals ◽

Anterior Teeth ◽

Sealing Ability ◽

Significant Difference

The aim of this study was to evaluate the sealing ability of three different canal filling techniques. Sixty-four roots of extracted human maxillary anterior teeth were prepared using ProTaper® rotary instruments. The specimens were then randomly divided into 3 experimental groups (n=16) and 2 control groups (n=8). The root canals were filled using cold lateral compaction (CLC group), continuous wave condensation technique using the Elements Obturation Unit® (EOU group), and ProTaper obturators (PT group). For the negative control group, 8 roots were filled using lateral compaction as in the CLC group, and the teeth were covered twice with a layer of nail varnish (NCG group). Another 8 roots were filled using lateral compaction, but without sealer, and these were used as the positive control (PCG group). A glucose leakage model was used for quantitative evaluation of microleakage for 24 hours and 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12 weeks. No significant difference in the cumulative amount of leakage was found between the three experimental groups at all observation times. The lateral condensation of cold gutta-percha can guarantee a similar seal of canal fillings as can be achieved by using thermal methods, in the round canals.

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Astragaloside positively regulated osteogenic differentiation of pre-osteoblast MC3T3-E1 through PI3K/Akt signaling pathway

Journal of Orthopaedic Surgery and Research ◽

10.1186/s13018-021-02690-1 ◽

2021 ◽

Vol 16 (1) ◽

Author(s):

Wei Bing Jing ◽

Hongjuan Ji ◽

Rui Jiang ◽

Jinlong Wang

Keyword(s):

Western Blot ◽

Osteogenic Differentiation ◽

Real Time ◽

Signaling Pathway ◽

Real Time Pcr ◽

Akt Signaling ◽

Calcium Deposition ◽

Control Group ◽

Akt Signaling Pathway ◽

Western Blot Assay

Abstract Background Osteoporosis is a widespread chronic disease characterized by low bone density. There is currently no gold standard treatment for osteoporosis. The aim of this study was to explore the role and mechanism of Astragaloside on osteogenic differentiation of MC3T3-E1 cells. Methods MC3T3-E1 cells were divided into control and different dose of Astragaloside (10, 20, 40, 50, and 60 μg/ml). Then, ALP and ARS staining were performed to identify the effects of Astragaloside for early and late osteogenic capacity of MC3T3-E1 cells, respectively. Real-time PCR and western blot were performed to assess the ALP, OCN, and OSX expression. PI3K/Akt signaling pathway molecules were then assessed by Western blot. Finally, PI3K inhibitor, LY294002, was implemented to assess the mechanism of Astragaloside in promoting osteogenic differentiation of MC3T3-E1 cells. Results Astragaloside significantly increased the cell viability than the control group. Moreover, Astragaloside enhanced the ALP activity and calcium deposition than the control groups. Compared with the control group, Astragaloside increased the ALP, OCN, and OSX expression in a dose-response manner. Western blot assay further confirmed the real-time PCR results. Astragaloside could significantly increase the p-PI3K and p-Akt expression than the control group. LY294002 partially reversed the promotion effects of Astragaloside on osteogenic differentiation of MC3T3-E1 cells. LY294002 partially reversed the promotion effects of Astragaloside on ALP, OCN, and OSX of MC3T3-E1 cells. Conclusion The present study suggested that Astragaloside promoted osteogenic differentiation of MC3T3-E1 cells through regulating PI3K/Akt signaling pathway.

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A Digital Microscopic Inspection of Dentinal Defects after Using Endodontic Retreatment Files

International Journal of Dentistry ◽

10.1155/2021/6661387 ◽

2021 ◽

Vol 2021 ◽

pp. 1-6

Author(s):

Ammar AbuMostafa ◽

Hala Almoqayyad ◽

Al-Omari Mohammad

Keyword(s):

Negative Control Group ◽

Positive Control ◽

Negative Control ◽

Control Group ◽

Filling Material ◽

Root Canals ◽

Root Canal Retreatment ◽

Significant Difference ◽

Digital Microscope ◽

Microscopic Inspection

Aim. The study aimed at evaluating the incidence of dentinal defects after root canal retreatment with ProTaper Universal retreatment (PTUR) and XP-endo Shaper and Finisher R (XP). Materials and Methods. Sixty extracted single-rooted human premolars were selected and divided into 4 groups of 15 teeth each. In the negative control group, the teeth were left unprepared. In the positive control group, the teeth were prepared with ProTaper Next and obturated with no further retreatment. In the PTUR and XP groups, the teeth were prepared and obturated followed by removal of the filling material at body temperature using PTUR and XP instruments, respectively. The roots were then sectioned at 3, 6, and 9 mm from the apex and observed under a digital microscope to detect defects. Results. PTUR group showed significantly higher ( p value <0.05) incidence of defects than the other groups. Comparison of no defects versus defects between groups in different areas of root canals demonstrated significant difference among the groups in the apical and cervical regions. Conclusion. Within the limitations of the present study, PTUR files created significantly more dentinal defects than XP files, with most of those defects at the cervical and apical areas of the root canals.

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TLR, RAGE, and HMGB1 in the Inflammatory Response in Ph(-) Myeloproliferative Neoplasm

Blood ◽

10.1182/blood-2020-134988 ◽

2020 ◽

Vol 136 (Supplement 1) ◽

pp. 31-32

Author(s):

Guanfang Shi ◽

Kiron Nair ◽

Preethi Ramachandran ◽

Chi Chen ◽

Ching Wong ◽

...

Keyword(s):

Inflammatory Response ◽

Real Time ◽

Inflammatory Cytokines ◽

Internal Control ◽

Real Time Pcr ◽

Mononuclear Cells ◽

Conflicts Of Interest ◽

Philadelphia Chromosome ◽

Myeloproliferative Neoplasm ◽

Significant Difference

Recent evidence of increased constitutional symptoms and inflammatory cytokines in Philadelphia chromosome negative (Ph (-)) MPN suggests that an inflammatory response is important in the pathogenesis of Ph (-) MPN. Toll-like receptors (TLR), Receptor for Advanced Glycation End products (RAGE) and High mobility group protein B1 (HMGB1) are the important pathways for the inflammatory response. All these three important pathway proteins were studied in MPN diseases in the current studies. Materials and Methods: TLR assay. TLR 2,3, 4, 7, 9 quantification was performed by immuno-staining of 1×106 mononuclear cells (peripheral blood) which were incubated with fluorescence-conjugated anti-TLR-2,3, 4, 7, 9 antibodies and assayed by flow cytometry. HMGB1assay:HMGB1 ELISA kit from Immuno-Biological Laboratories, Inc. (IBL-America) were used. The plasma samples were diluted four times with the provided sample dilution buffer, and assayed in duplicate according to the manufacturer's suggestion. RAGE (RT-PCR) Assay: Total RNA was extracted from normal control or patient mononuclear cells. Predesigned primers for RAGE, and internal control genes were ordered from Qiagen (Germantown, MD). Real-time PCR was performed using SsoAdvanced™ Universal SYBR® Green Supermix (Bio-Rad, Hercules, CA) on Bio-Rad iQ5 Multicolor Real-Time PCR Detection System. At least three house-keeping genes (ribosomal protein L4, TATA box binding protein, and tubulin-α 1b) were used as normalization controls. The expression of RAGE were compared with each internal control. Average of three was used to calculate the ratio of final patient to normal Results: Total of 97 patients with MPN were studied 1) TLR: TLR 3,7,9 was not significantly different from controls. But TLR 2 was significantly increased in both PV, as well as in the MPN group when PV, ET and MF were grouped together as MPN (Fig A). TLR 4 was not significantly increased in PV, ET, MF individually but was found to be significantly increased than the controls, when they are grouped together as MPN (Fig B). 2) RAGE: No significant difference was found between ET, PV, MF individually or when they were grouped together as MPN than the controls (Fig C). 3) HMGB1: No significant difference was seen between ET, PV, MF or when they were grouped as MPN (Fig D). Conclusion: Current study suggests that TLR pathway especially TLR2, and to a lesser extent TLR4 are the important pathways for inflammatory response with increased inflammatory cytokines in MPN, while HMGB1 and RAGE pathways were not different from controls. Figure Disclosures No relevant conflicts of interest to declare.

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Diversity of KIR genes and their HLA-C ligands in Ugandan populations with historically varied malaria transmission intensity

10.21203/rs.3.rs-45487/v2 ◽

2021 ◽

Author(s):

Stephen Tukwasibwe ◽

James A. Traherne ◽

Olympe Chazara ◽

Jyothi Jayaraman ◽

John Trowsdale ◽

...

Keyword(s):

Infectious Diseases ◽

Malaria Transmission ◽

Real Time ◽

High Throughput ◽

Real Time Pcr ◽

Association Studies ◽

Transmission Intensity ◽

Malaria Transmission Intensity ◽

Significant Difference ◽

Pcr Method

Abstract Background: Malaria is one of the most serious infectious diseases in the world. The malaria burden is greatly affected by human immunity, and immune responses vary between populations. Genetic diversity in KIR and HLA-C genes, which are important in immunity to infectious diseases, is likely to play a role in this heterogeneity. Several studies have shown that KIR and HLA-C genes influence the immune response to viral infections, but few studies have examined the role of KIR and HLA-C in malaria infection, and these have used low-resolution genotyping. The aim of this study was to determine whether genetic variation in KIR and their HLA-C ligands differ in Ugandan populations with historically varied malaria transmission intensity using more comprehensive genotyping approaches.Methods: High throughput multiplex quantitative real-time PCR method was used to genotype KIR genetic variants and copy number variation and a high-throughput real-time PCR method was developed to genotype HLA-C1 and C2 allotypes for 1,344 participants, aged 6 months to 10 years, enrolled from Ugandan populations with historically high (Tororo District), medium (Jinja District) and low (Kanungu District) malaria transmission intensity. Results: The prevalence of KIR3DS1, KIR2DL5, KIR2DS5 and KIR2DS1 genes was significantly lower in populations from Kanungu compared to Tororo (7.6% vs. 13.2%: p=0.006, 57.2% vs. 66.4%: p=0.005, 33.2% vs. 46.6%: p<0.001 and 19.7% vs. 26.7%: p=0.014 respectively) or Jinja (7.6% vs.18.1%: p<0.001, 57.2% vs. 63.8%: p=0.048, 33.2% vs. 43.5%: p=0.002 and 19.7% vs. 30.4%: p<0.001 respectively). The prevalence of homozygous HLA-C2 was significantly higher in populations from Kanungu (31.6%) compared to Jinja (21.4%), p=0.043, with no significant difference between Kanungu and Tororo (26.7%), p=0.296. Conclusions: The KIR3DS1, KIR2DL5, KIR2DS5 and KIR2DS1 genes may partly explain differences in transmission intensity of malaria since these genes have been positively selected for in places with historically high malaria transmission intensity. The high-throughput multiplex real-time HLA-C genotyping PCR method developed will be useful in disease association studies involving large cohorts.

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