With the increasing availability of genetic tests, more doctors are offering and ordering such tests for their patients. Ordering a genetic test appears to be a simple process of filling in paperwork, drawing 3 mL of blood in an ethylenediaminetetraacetic acid tube and receiving a test report. This is identical to sending off a full blood count. However, it is far more complex than that. There are many potential pitfalls, as shown by the increasing number of complaints and lawsuits filed against doctors and allied health staff. Furthermore, clinical genetics involves more than just ordering tests; in fact, focusing on genetic tests alone is a potential pitfall. In this review, we discuss the common pitfalls in clinical genetics and how doctors can avoid these pitfalls to ensure patient safety and to safeguard their practice.
Humans ; Edetic Acid ; Fenbendazole ; Patient Safety ; Physicians

OBJECTIVE: To investigate the value of ⁶⁸Ga-labeled prostate specific membrane antigen (PSMA) PET/CT for assessing tumor load in primary lesions for risk stratification and predicting metastasis of newly diagnosed prostate cancer (PCa). METHODS: We retrospectively analyzed the data of 36 patients (mean age 71.3 ± 8.6 years, range 56 to 89 years) with newly diagnosed PCa undergoing ⁶⁸Ga-PSMA-I&T PET/CT from June 2018 to July 2019. SUVmax and SUVmean of the primary lesions were measured, and the primary PSMA tumor volume (PSMA-TV) and total lesion PSMA (TL-PSMA) were automatically measured and calculated in all the patients. The correlations of primary SUVmax, PSMA-TV, and TL-PSMA with PSA and Gleason score (GS) were analyzed, and SUVmax, PSMA-TV and TL-PSMA of the primary lesions were compared among different PCa subgroups. RESULTS: SUVmax, PSMA-TV and TL-PSMA of the primary lesions were all correlated with PSA and GS (P < 0.05). PCa subgroup analysis showed that SUVmax, PSMA-TV and TL-PSMA were all significantly higher in patients with PSA >20 ng/mL than in those with PSA ≤20 ng/mL (P < 0.001), and were higher in patients with a GS ≥8 than in those with a GS ≤7 (P < 0.001). PSMA-TV and TL-PSMA were significantly higher in patients with tumor metastasis than in those without metastasis (P < 0.001), while SUVmax did not differ significantly with tumor metastasis. SUVmax (P=0.002), PSMA-TV (P < 0.001), and TL-PSMA (P < 0.001) were all significantly higher in high-risk group than in low-to moderate-risk group. CONCLUSION PSMA-TV and TL-PSMA of ⁶⁸Ga-PSMA-I&T PET/CT have potential value in predicting risk stratification and metastasis of newly diagnosed PCa.
Aged ; Aged, 80 and over ; Edetic Acid ; Gallium Isotopes ; Gallium Radioisotopes ; Humans ; Male ; Middle Aged ; Oligopeptides ; Positron Emission Tomography Computed Tomography ; Prostate-Specific Antigen ; Prostatic Neoplasms/pathology* ; Retrospective Studies ; Tumor Burden

The aim of this study was to compare the efficiency of four final irrigation protocols in smear layer removal and bacterial inhibition in root canal systems. Thirty roots inoculated with Enterococcus faecalis were prepared with ProTaper Universal files. The teeth were disinfected by conventional needle irrigation, sonic agitation using the EndoActivator device, passive ultrasonic irrigation, or an M3 Max file. Teeth with no root canal preparation served as blank controls for the establishment of the infection baseline. Teeth with preparation but no final irrigation served as a post-instrumentation baseline. After the final irrigation, the teeth were sectioned in half. One half of each tooth was examined by scanning electron microscopy (SEM) to assess smear layer removal using a five-point scale. The other half was examined by confocal laser scanning microscopy (CLSM) using the LIVE/DEAD BackLight bacterial viability kit to evaluate the depth of bacterial survival in dentinal tubules. SEM analysis revealed no significant difference in smear layer removal throughout the whole canal among the EA, PUI, and M3 Max groups (P > 0.05). CLSM revealed that PUI achieved the greatest bacterial inhibition depth in the coronal ((174.27 ± 31.63) μm), middle ((160.94 ± 37.77) μm), and apical ((119.53 ± 28.49) μm) thirds of the canal (all P < 0.05 vs. other groups). According to this comprehensive SEM and CLSM evaluation, PUI appears to have the best infection control ability in root canal systems.
Dental Pulp Cavity ; Edetic Acid ; Humans ; Microscopy, Electron, Scanning ; Root Canal Irrigants ; Root Canal Preparation ; Smear Layer ; Sodium Hypochlorite

BACKGROUND: Extraction of cell-free DNA (cfDNA) is a key step for determining the quality of cfDNA-related molecular diagnostics. We evaluated the effect of sample containers and sample storage conditions on cfDNA extraction. METHODS: The cfDNA extraction using the MagMAX Cell-Free DNA Isolation Kit from five healthy controls and five lung cancer patients was evaluated according to the type of sample container and storage conditions: K2-EDTA container, <1, 6, 24, and 48 hr storage at 4℃ after immediate plasma separation; and Cell-Free DNA BCT container, <1, 3, 7, and 14 days stored at room temperature. Mutation analysis of EGFR exons 18–21 was performed. To assess the effect of a delay in centrifugation, EDTA whole blood samples from five healthy individuals were stored at 4℃ for 6, 12, and 24 hr before plasma separation. RESULTS: There was no significant difference in the amount and nucleic acid size of cfDNA in both controls and patients with cancer when EDTA plasma was stored at 4℃ up to 48 hr. The amount and size of cfDNA in the BCT container were not different up to 7 days; however, the 14-day sample showed an increase in cfDNA concentration due to genomic DNA contamination. EGFR mutations were detected on EDTA containers up to 48 hr and with BCT containers up to 14 days. When EDTA whole blood was stored at 4℃ and plasma separation was delayed, the cfDNA concentration increased from 24 hr. CONCLUSIONS: The cfDNA extraction was affected by the sample containers and storage conditions.
Biopsy ; Centrifugation ; DNA Contamination ; DNA ; Edetic Acid ; Exons ; Humans ; Lung Neoplasms ; Pathology, Molecular ; Plasma

OBJECTIVE: This work aims to uncover the promoting effect of 17% ethylenediaminetetraacetic acid (EDTA) irrigation on the dentin adhesion of Enterococcus faecalis (E. faecalis). METHODS: Forty-eight half split samples and twelve dentin slices were randomly divided into three experimental groups and one control group. The experimental groups and the control group were soaked with EDTA in different time lengths and with normal saline, respectively. E. faecalis was inoculated, and its dentin adhesion was measured via scanning electron microscopy, confocal laser scanning microscopy (CLSM), colony forming unit counts, and histological Gram staining. RESULTS: According to histological Gram staining, the depth showed no statistically significant differences between 1 min group and the control group, 1 min group and 3 min groups (P>0.05). E. faecalis intruded in the dentine tubules (measured by CLSM), and the thickness of the biofilm on the dentin surface and the colony numbers of experimental groups were greater than those of the control group (P<0.05). The differences between the three experimental groups were statistically signi-ficant (P<0.05). CONCLUSIONS EDTA (17%) irrigation can promote E. faecalis adhesion to dentin. This adhesion would in turn prolong EDTA treatment time.
Biofilms ; Dentin ; Edetic Acid ; Enterococcus faecalis ; Microscopy, Confocal ; Root Canal Irrigants ; Sodium Hypochlorite

OBJECTIVE: This study aimed to evaluate the effectiveness of XP-endo Finisher (XPF) file and passive ultrasonic irrigation (PUI) in the smear layer removal of the root canal. METHODS: A total of 60 human single-rooted premolars were selected and decoronated to standardize their canal length to 16 mm. Tooth samples were prepared using a S3 rotary system to prepare root canal with the file size of 3S and then randomly divided into 6 groups according to the final irrigation protocol, as follows: XPF 3 mL of 3% NaOCl for 1 min (group A); XPF 3 mL of 3% NaOCl for 1 min, followed by 4 mL of 17% ethylene diamine tetraacetic acid (EDTA) for 1 min (group B); PUI of 3 mL of 3% NaOCl for 1 min (group C); PUI of 3 mL of 3% NaOCl for 1 min, followed by 4 mL of 17% EDTA for 1 min (group D); 3 mL of 3% NaOCl for 1 min by using a syringe and a 30 G side-vented needle (group E); and 3 mL of 3% NaOCl for 1 min by using a syringe and a 30 G side-vented needle, followed by 4 mL of 17% EDTA for 1 min (group F). After the completion of the root canal preparation, the teeth were split into two longitudinally. The mean numbers of the visible open dentinal tubules in the apical and middle thirds of the root canals were evaluated via scanning electron microscope. RESULTS: The whole surfaces of the root canals in groups A, C, and E were covered by a smear layer. Groups A and C possessed significantly higher number of visible open dentinal tubules than in group E (P<0.05), with statistically insignificant difference between groups A and C (P>0.05). The apical third of the samples in groups B and D and in the middle thirds of canals in group F exhibited a small amount of smear layer, and the dentinal tubules were open or semi-open. The root canal surfaces in the apical third of the samples in group F were covered by a smear layer, and the dentinal tubules were sealed or semisealed. The smear layers in the middle third of the samples in groups B and D were removed, and the dentinal tubules were more visibly open than those of the four other groups (P<0.05). The difference between groups B and D were statistically insignificant (P>0.05). CONCLUSIONS The difference between XPF and PUI in terms of the smear layer removal of the root canals was insignificant. Hence, XPF, as a new irrigation agitation technique, can aid in improving smear layer removal.
Dental Pulp Cavity ; Edetic Acid ; Humans ; Microscopy, Electron, Scanning ; Root Canal Irrigants ; Root Canal Preparation ; Smear Layer ; Sodium Hypochlorite

The Luminex-based single antigen bead (SAB) assay is widely used to detect HLA antibody in transplant recipients. However, one limitation of the SAB assay is the prozone effect, which occurs mostly as a result of complement interference. We investigated the efficacy of EDTA treatment for overcoming the prozone effect and predicting C1q binding of HLA antibody. We subjected 27 non-treated (naïve) and EDTA-treated serum samples from highly sensitized patients to IgG-SAB assays, and we confirmed the prozone effect in 53% and 31% of class I and class II antibody tests, respectively, after EDTA treatment. When we conducted additional assays after dithiothreitol treatment and serum dilution, EDTA was the most efficacious in eliminating the prozone effect. Reducing the prozone effect by EDTA treatment strengthened the correlation between IgG mean fluorescence intensity (MFI) and C1q MFI values (ρ=0.825) as compared with the naïve sera (ρ=0.068). Although C1q positivity was dependent on the concentration of HLA antibody in EDTA-treated sera, the correlations varied individually. Overall, our results confirmed the efficacy of EDTA treatment for overcoming the prozone effect. EDTA treatment showed a positive effect on the correlation between IgG MFI and C1q MFI values.
Complement System Proteins ; Dithiothreitol ; Edetic Acid ; Fluorescence ; Humans ; Immunoglobulin G ; Transplant Recipients

OBJECTIVES: To achieve pulp-dentin complex regeneration with tissue engineering, treatment efficacies and safeties should be evaluated using in vivo orthotopic transplantation in a sufficient number of animals. Mice have been a species of choice in which to study stem cell biology in mammals. However, most pulp-dentin complex regeneration studies have used large animals because the mouse tooth is too small. The purpose of this study was to demonstrate the utility of the mouse tooth as a transplantation model for pulp-dentin complex regeneration research. MATERIALS AND METHODS: Experiments were performed using 7-week-old male Institute of Cancer Research (ICR) mice; a total of 35 mice had their pulp exposed, and 5 mice each were sacrificed at 1, 2, 4, 7, 9, 12 and 14 days after pulp exposure. After decalcification in 5% ethylenediaminetetraacetic acid, the samples were embedded and cut with a microtome and then stained with hematoxylin and eosin. Slides were observed under a high-magnification light microscope. RESULTS: Until 1 week postoperatively, the tissue below the pulp chamber orifice appeared normal. The remaining coronal portion of the pulp tissue was inflammatory and necrotic. After 1 week postoperatively, inflammation and necrosis were apparent in the root canals inferior to the orifices. The specimens obtained after experimental day 14 showed necrosis of all tissue in the root canals. CONCLUSIONS: This study could provide opportunities for researchers performing in vivo orthotopic transplantation experiments with mice.
Animals ; Biology ; Dental Pulp Cavity ; Dental Pulp Necrosis ; Edetic Acid ; Eosine Yellowish-(YS) ; Hematoxylin ; Humans ; Inflammation ; Male ; Mammals ; Mice ; Necrosis ; Pulpitis ; Regeneration ; Safety ; Stem Cells ; Tissue Engineering ; Tooth

OBJECTIVE: To investigate the effect of smear layer on apical sealing ability in teeth obturated with mineral trioxide aggregate (MTA) Plus as retrofilling materials. METHODS: Fifty freshly extracted maxillary anterior teeth or premolars with single root canal were used in this study. All teeth were instrumented to master apical point 60# by using the stepback technique, obturated with lateral condensation technique, and then apical resected. A rootend cavity was then instrumented with an ultrasonic diamond-coated tip. Then the selected teeth were randomly and equally divided into two groups (n=25). In the experimental group (smear-), the teeth were irrigated with 0.17 g/L ethylenediaminetetraacetic acid (EDTA) to remove smear layer on the root-end cavity wall; in the control group (smear+), the teeth were irrigated with physiological saline. Five teeth were extracted to evaluate the cleanliness of root end cavity walls under a videomicroscope, respectively. The scanning electron microscope (SEM) evaluation was also performed for the presence of smear layer and open tubule. For the additional 40 teeth, the root-end cavities were filled with MTA Plus. The quantitative apical leakage of each teeth was evaluated by measuring the concentration of leaked sucrose in apical reservoir on 1, 7, 14, 21, 28, 35, 42, 49 and 56 days, respectively. The samples were stored at 37 °C and 100% humidity for 56 days. Statistical analysis was done with ANOVA for repeated measurement design data. RESULTS: Removal of the smear layer did not cause significantly less apical leaked sucrose than that when the smear layer was left intact for 56 days (P>0.05). There were statistically significant differences at the concentration of leaked sucrose among different observation time points (P<0.05). CONCLUSION It may be concluded that removing the smear layer may not be necessary in root-end cavities filled with MTA Plus.
Aluminum Compounds ; Calcium Compounds ; Dental Leakage ; Drug Combinations ; Edetic Acid ; Oxides ; Root Canal Filling Materials ; Root Canal Irrigants ; Root Canal Preparation ; Root Canal Therapy ; Silicates ; Smear Layer ; Sucrose/pharmacokinetics*

BACKGROUND: Antibodies specific to human neutrophil antigen (HNA), especially HNA-2, are implicated in various conditions, including neonatal alloimmune neutropenia, febrile non-hemolytic transfusion reactions, and transfusion-related acute lung injury. The distribution of the HNA-2 phenotype frequencies in the Thai population remains unknown. This study aimed to investigate HNA-2 phenotype frequencies in Thai blood donors and to compare the relationships of sex and age with HNA-2 expression. METHODS: EDTA blood samples were collected from 220 unrelated healthy Thai blood donors, including 150 males and 70 females, with ages ranging from 20 to 57 years. Polymorphonuclear cells were isolated and stained with monoclonal antibodies clone MEM-166 and clone 2D1, which are specific to human CD177 (HNA-2) and CD45, respectively. HNA-2 expression according to sex and age was analyzed by flow cytometry. RESULTS: Among the 220 donors, HNA-2-positive and HNA-2-null-phenotype frequencies were 0.995 and 0.005, respectively. Mean antigen expression was significantly higher in women (71.01±15.46%) than in men (64.59±18.85%; P < 0.05). No significant differences in HNA-2 expression were found between different age groups. HNA-2 phenotype frequencies were similar to those in Asian, African, American, and Brazilian populations, but were significantly different from those in eastern Japanese, Korean, and French populations (P < 0.001). CONCLUSIONS: This is the first report of HNA-2 phenotype frequencies in a Thai population, and the data will be helpful in predicting the risk of HNA-2 alloimmunization and in recruiting granulocyte panel donors.
Acute Lung Injury ; Antibodies ; Antibodies, Monoclonal ; Asian Continental Ancestry Group* ; Blood Donors* ; Clone Cells ; Edetic Acid ; Febrile Neutropenia ; Female ; Flow Cytometry ; Granulocytes ; Humans ; Male ; Neutrophils ; Phenotype* ; Tissue Donors ; Transfusion Reaction