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BACKGROUND: Apical surgery with standard retrograde maneuvers may be challenging in certain cases. Simplifying apical surgery to reduce operating time and streamline retrograde manipulation is an emerging need in clinical endodontics. AIM OF THE STUDY: The aim of the study was to compare the bacterial sealing ability of a calcium silicate-based sealer with the single cone technique combined with root end resection only, and calcium silicate-based sealer as a retrograde filling versus MTA retrofilling, and to analyze bacterial viability using confocal laser scanning microscope (CLSM). MATERIALS AND METHODS: In this in vitro experimental study, 50 extracted human maxillary incisor teeth were instrumented and randomly divided into five groups: three experimental groups, a positive control group, and a negative control group (n = 10/group). In the experimental groups, the roots were obturated using the single cone technique (SCT) and a calcium silicate-based sealer. In group 1, the roots were resected 3 mm from the apex with no further retrograde preparation or filling. In groups 2 and 3, the roots were resected, retroprepared, and retrofilled with either a calcium silicate-based sealer or MTA, respectively. Group 4 (positive control) was filled with a single gutta-percha cone without any sealer. In group 5 (negative control), the canals were left empty, and the roots were sealed with wax and nail varnish. A bacterial leakage model using Enterococcus faecalis was employed to assess the sealing ability over a 30-day period, checking for turbidity and analyzing colony forming units (CFUs) per milliliter. Five specimens from each group were examined using CLSM for bacterial viability. Data for the bacterial sealing ability were statistically analyzed using chi-squared and Kruskal-Wallis tests. RESULTS: The three experimental groups did not show significant differences in terms of bacterial leakage, or bacterial counts (CFUs) (P > 0.05). However, significant differences were observed when comparing the experimental groups to the positive control group. Notably, the calcium silicate-based sealer, when used as a retrofilling, yielded the best sealing ability. CLSM imaging revealed viable bacterial penetration in all the positive control group specimens while for the experimental groups, dead bacteria was the prominent feature seen. CONCLUSION: Within the limitations of this study, it could be concluded that the bacterial sealing ability of calcium silicate-based sealer with the single cone technique combined with root end resection only and calcium silicate-based sealer as a retrograde filling were comparable with MTA retrofilling during endodontic surgical procedures.

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PURPOSE: The current study aimed at assessing vitamin D level and vitamin D receptor polymorphism in hypothyroid Egyptian patients and its effect on hypothyroidism and thyroid morphology, also to find a causal relation between vitamin D and hypothyroidism. METHODS: This case-control study was conducted on 35 hypothyroid patients and 35 matched unrelated healthy controls. Total serum 25-hydroxyvitamin D3 and thyroid antibodies were measured using a human ELISA kit. Genotyping was performed by using real-time PCR. HOMA-IR was also calculated (fasting insulin in mIU/L × fasting glucose in mg/dL/405). All subjects were assessed for thyroid morphology by thyroid ultrasonography. RESULTS: Vitamin D level was lower in hypothyroid patients than in control subjects. Vitamin D was also inversely related to TSH, HOMA-IR, and levels of anti-TG and anti-TPO. VDR polymorphism (Fok1 and Apa1) had no relation to TSH or vitamin D levels in both patients and control groups. Low vitamin D levels were associated with increased thyroid vascularity and nodularity; furthermore, vitamin D was inversely proportional to thyroid gland volume. Correlation of HOMA-IR with the levels of both anti-TG and anti-TPO in the 70 subjects proved that HOMA-IR was positively correlated to both antibodies. CONCLUSION: This study confirmed the association of vitamin D deficiency with hypothyroidism, thyroid autoimmunity, increased volume, nodularity, and vascularity of thyroid gland in hypothyroid patients as well as increased HOMA-IR. It proved the association between HOMA-IR and thyroid autoimmunity. The study proved no association between VDR polymorphisms (Fok1 and Apa1) with either vitamin D levels or TSH levels.

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Quantum dots (QDs) are semiconductor nanocrystals that possess unique optical properties including broad-range excitation, size-tunable narrow emission spectra and high photostability, giving them considerable value in various biomedical applications. The size and composition of QDs can be varied to obtain the desired emission properties and make them amenable to simultaneous detection of multiple targets. Furthermore, numerous surface functionalizations can be used to adapt QDs to the needed application. The successful use of QDs has been reported in the areas of in vitro diagnostics and imaging. There is also potential for multimodal applications for simultaneous imaging. Toxicity issues are still a prime concern with regards to in vivo applications on account of the toxic constituents of QDs.

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In order to meet the challenges of effective healthcare, the clinical laboratory is constantly striving to improve testing sensitivity while reducing the required time and cost. Gold nanoparticles (AuNPs) are proposed as one of the most promising tools to meet such goals. They have unique optophysical properties which enable sensitive detection of biomarkers, and are easily amenable to modification for use in different assay formats including immunoassays and molecular assays. Additionally, their preparation is relatively simple and their detection methods are quite versatile. AuNPs are showing substantial promise for effective practical applications and commercial utilization is already underway. This article covers the principles of preparation of AuNPs and their use for development of different diagnostic platforms.

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CONTEXT: Correction factors have been proposed for estimating true potassium concentrations in blood samples with evidence of in vitro hemolysis. OBJECTIVE: We used 2 different models of true (ie, nonsimulated) in vitro hemolysis to evaluate the clinical utility of correction factors for estimating potassium concentrations in samples with evidence of in vitro hemolysis. DESIGN: Potassium correction factors were derived using 2 different models. In model 1, potassium and plasma hemoglobin were measured with the Hitachi 747 analyzer in 132 paired blood samples, with each pair consisting of 1 sample with evidence of hemolysis and 1 without, collected during the same phlebotomy procedure. The change in measured potassium concentration was plotted versus the change in plasma hemoglobin concentration for each pair of samples. In model 2, the potassium levels of 142 784 blood samples and the corresponding hemolytic index values were measured with the Beckman LX20 analyzer. Potassium concentrations at the 10th, 25th, 50th, 75th, and 90th percentiles were calculated for each hemolysis index category. RESULTS: From our 2 models, we derived correction factors expressing an increase in potassium concentration of 0.51 and 0.40 mEq/L for every increase in plasma hemoglobin concentration of 0.1 g/dL. These correction factors are much higher than those reported in studies that simulated in vitro hemolysis by freeze-thaw lysis or osmotic disruption of whole blood. CONCLUSIONS: Use of correction factors for estimating the true potassium concentration in samples with evidence of in vitro hemolysis is not recommended. Derivation of correction factors by using samples with nonsimulated in vitro hemolysis suggests that the actual increase in potassium in hemolyzed samples is much higher than that reported in previous studies that produced hemolysis with artificial means.

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There is a constant need to improve the performance of current diagnostic assays as well as develop innovative testing strategies to meet new testing challenges. The use of nanoparticles promises to help promote in vitro diagnostics to the next level of performance. Quantum dots (QDs), gold nanoparticles (AuNPs), and superparamagnetic nanoparticles are the most promising nanostructures for in vitro diagnostic applications. These nanoparticles can be conjugated to recognition moieties such as antibodies or oligonucleotides for detection of target biomolecules. Nanoparticles have been utilized in immunoassays, immunohistochemistry, DNA diagnostics, bioseparation of specific cell populations, and cellular imaging. Nanoparticle-based diagnostics may open new frontiers for detection of tumours, infectious diseases, bio-terrorism agents, and neurological diseases, to name a few. More work is necessary to fully optimize use of nanoparticles for clinical diagnosis and to resolve some concerns regarding potential health and environmental risks related to their use. However, we envision further developments of nanoparticle-based diagnostics will yield unique assays with enhanced sensitivity and multiplexing capability for the modern clinical laboratory.

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Gene doping is the newest threat to the spirit of fair play in sports. Its concept stemmed out from legitimate gene therapy trials, but anti-doping authorities fear that they now may be facing a form of doping that is virtually undetectable and extremely appealing to athletes. This paper presents studies that generated mouse models with outstanding physical performance, by manipulating genes such as insulin-like growth factor 1 (IGF-1) or phosphoenolpyruvate carboxykinase (PEPCK), which are likely to be targeted for gene doping. The potential transition from super mice to super athletes will also be discussed, in addition to possible strategies for detection of gene doping.

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Gene doping, the abuse of gene therapy for illicit athletic enhancement, is perceived as a coming threat and is a prime concern to the anti-doping community. This doping technique represents a significant ethical challenge and there are concerns regarding its safety for athletes. This article presents the basics of gene doping, potential strategies for its detection and the role of promising new technologies in aiding detection efforts. These include the use of lab-on-a-chip techniques as well as nanoparticles to enhance the performance of current analytical methods and to develop new doping detection strategies.

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The quest for athletic excellence holds no limit for some athletes, and the advances in recombinant DNA technology have handed these athletes the ultimate doping weapons: recombinant proteins and gene doping. Some detection methods are now available for several recombinant proteins that are commercially available as pharmaceuticals and being abused by dopers. However, researchers are struggling to come up with efficient detection methods in preparation for the imminent threat of gene doping, expected in the 2008 Olympics. This Forum article presents the main detection strategies for recombinant proteins and the forthcoming detection strategies for gene doping as well as the prime analytical challenges facing them.

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Quantum dots (QDs) are among the most promising items in the nanomedicine toolbox. These nanocrystal fluorophores have several potential medical applications including nanodiagnostics, imaging, targeted drug delivery, and photodynamic therapy. The diverse potential applications of QDs are attributed to their unique optical properties including broad-range excitation, size-tunable narrow emission spectra, and high photostability. The size and composition of QDs can be varied to obtain the desired emission properties a makes them amenable for simultaneous detection of multiple targets. Also, numerous surface functionalizations can be used to adapt QDs to the needed application. Recent reports have shown successful use of QDs in various medical applications. With respect to in vivo applications, caution must be exercised with QDs due to their toxic components. Development of appropriate health and safety regulations and resolution of intellectual property issues are necessary for commercialization. In light of these obstacles however, QDs appear to be too valuable to nanomedicine to dismiss, and will eventually come into routine practical use.

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BACKGROUND: The use of nanotechnologies for diagnostic applications shows great promise to meet the rigorous demands of the clinical laboratory for sensitivity and cost-effectiveness. New nanodiagnostic tools include quantum dots (QDs), gold nanoparticles, and cantilevers. QDs, which are the most promising nanostructures for diagnostic applications, are semiconductor nanocrystals characterized by high photostability, single-wavelength excitation, and size-tunable emission. QDs and magnetic nanoparticles can be used for barcoding of specific analytes. Gold and magnetic nanoparticles are key components of the bio-barcode assay, which has been proposed as a future alternative to the PCR. METHODS: We examined articles published over the past 10 years investigating the use of QDs, gold nanoparticles, cantilevers, and other nanotechnologies in promising diagnostic applications. RESULTS: Several nanodiagnostic assays have been developed, including a QD-based assay capable of detecting biotinylated prostate-specific antigen (PSA) at 0.38 ng/L, a bio-barcode assay capable of detecting 30 amol/L PSA in a 10-microL sample, and another able to detect 50 molecules of the Alzheimer marker amyloid beta-derived diffusible ligand in 10 microL of cerebrospinal fluid. CONCLUSIONS: Nanodiagnostics promise increased sensitivity, multiplexing capabilities, and reduced cost for many diagnostic applications as well as intracellular imaging. Further work is needed to fully optimize these diagnostic nanotechnologies for clinical laboratory setting and to address the potential health and environmental risks related to QDs.

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Advances in recombinant DNA technology have created one of the most powerful weapons in the current doping arsenal: recombinant proteins [Sweeney HL. Gene doping. Sci Am 2004;291:62-9; Unal M, Ozer Unal D. Gene doping in sports. Sports Med 2004;34:357-62]. Recombinant erythropoietin (EPO) and human growth hormone (hGH) are currently being abused but are fortunately detectable either directly by employing isoelectric focusing and immunoassays or indirectly by assessing changes in selected hematopoietic parameters. The detection is technically demanding due to the extent of similarity between the recombinant proteins and their endogenous counterparts. Another issue facing detection efforts is the speed and conditions at which blood samples are collected and analyzed in a sports setting. Recently, gene doping, which stemmed out of legitimate gene therapy trials, has emerged as the next level of doping. Erythropoietin (EPO), human growth hormone (hGH), insulin-like growth factor-1 (IGF-1), peroxisome proliferator-activated receptor-delta (PPAR delta), and myostatin inhibitor genes have been identified as primary targets for doping. Sports clinical scientists today are racing against the clock because assuring the continued integrity of sports competition depends on their ability to outpace the efforts of dopers by developing new detection strategies.

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