Removal of fiber posts using conventional versus guided endodontics: a comparative study of dentin loss and complications.

OBJECTIVES: To compare the efficacy of fiber post removal using conventional (CONV) versus guided endodontics (GE) in terms of dentin loss, residual resin material, procedural errors, and working time in vitro. MATERIAL AND METHODS: Ninety human central incisors were root-filled and scanned by micro-computed tomography (CT), then restored with fiber posts and composite. Twenty-four sets of teeth with up to four human maxillary central incisors were fabricated and divided into three groups: conventional post removal by a general dentist (CG) or endodontology specialist (CS) and guided endodontics (GE) by a general dentist, yielding 30 teeth per operator and group. After treatment, the prepared access cavities were volumetrically assessed by micro-CT. Statistical significance was evaluated by one-way analysis of variance followed by post hoc comparisons with Tukey's HSD test and Pearson's chi-squared test for independence. RESULTS: Both CONV and GE resulted in dentin loss and residual resin material. CS resulted in more dentin loss and less residual resin material than CG and GE (p < .05). All groups had some deviations from the original root canal but no perforations. The shortest working time was observed in the GE group. CONCLUSIONS: Compared to the conventional freehand technique, GE resulted in significantly less radicular dentin loss, a few deviations but no perforations. CLINICAL RELEVANCE: Guided endodontics can improve the speed and safety of fiber post removal without root perforation.

People living with HIV have low trabecular bone mineral density, high bone marrow adiposity, and poor trabecular bone microarchitecture at the proximal femur.

People living with HIV (PLWH) have increased risk of osteoporosis and fractures. We assessed the proximal femur of PLWH and age-matched seronegative controls using quantitative computed tomography and magnetic resonance imaging. Results suggest that the trabecular compartment is compromised at fracture-prone regions in the proximal femur of PLWH. INTRODUCTION: People living with HIV (PLWH) have increased risk of osteoporosis and fractures. However, studies assessing the main determinants of bone strength in the proximal femur exclude this vulnerable population. We assessed the proximal femur of 40 PLWH and 26 age-matched seronegative controls using quantitative computed tomography and magnetic resonance imaging. METHODS: We examined cortical volumetric bone mineral density (Ct.vBMD), trabecular vBMD (Tb.vBMD), cortical thickness (Ct.Th), bone marrow adiposity (BMA), and trabecular number, separation, and bone volume fraction. Parametric comparisons between the two groups were made for the femoral head, femoral neck, trochanter, and total hip using linear regression adjusting for several covariates, including metrics of body composition. In addition, we investigated the associations of BMA with Tb.vBMD and trabecular microarchitecture with Spearman's rank partial correlations. RESULTS: PLWH had lower Tb.vBMD and deteriorated trabecular microarchitecture in the femoral neck, trochanter and total hip, and elevated BMA in the femoral head, femoral neck, and total hip. Ct.vBMD and Ct.Th were not significantly different between the two groups. BMA was significantly associated with lower Tb.vBMD and deteriorated trabecular microarchitecture in both groups albeit at different femoral regions. CONCLUSIONS: Our findings suggest that the trabecular, and not the cortical, compartment is compromised in the proximal femur of PLWH. The observed impairments in fracture-prone regions in PLWH indicate lower femoral strength and suggest higher fracture risk. The inverse associations of BMA with trabecular bone density and microarchitecture quality agree with findings at other anatomic sites and in other populations, suggesting that excess BMA possibly due to a switch from the osteoblast to the adipocyte lineage may be implicated in the pathogenesis of bone fragility at the femur in PLWH.

A critical evaluation of the material properties and clinical suitability of in-house printed and commercial tooth replicas for endodontic training.

AIM: To assess the suitability of several 3D-printed resins for the manufacturing of tooth replicas for endodontic training in comparison with commercially available replicas by analysing the properties of the materials and comparing them with real teeth during endodontic training. METHODOLOGY: Tooth replicas were 3D-printed using four resins (NextDent Model, NextDent C&B, V-Print ee and Vero White Plus) and compared with two commercially available products (VDW and Smile Factory) as well as extracted human teeth. Martens hardness, indentation modulus and radiopacity were investigated on these tooth replicas. Experienced dentists evaluated the suitability of the replicas for endodontic training by comparing them with real teeth in terms of appearance, anatomy, radiopacity, similarity to dentine during access opening, canal gauging and canal instrumentation. Data were analysed using the Kolmogorov-Smirnov and Mann-Whitney U-test. RESULTS: The greatest hardness values were recorded for human dentine (P < 0.001), followed by V-Print ee and the commercial tooth replica of Smile Factory. The greatest radiopacity was associated with VOC and dentine (P < 0.001) in comparison with the other materials tested. The appearance of the in-house printed tooth replicas was subjectively evaluated by the dentists as being more realistic than the commercially available products. No differences between the replicas was detected during mechanical instrumentation of root canals. CONCLUSION: None of the tooth replicas were able to simulate human dentine from the perspectives evaluated. V-Print ee had radiopacity comparable with dentine, but its hardness was not comparable with dentine.

Technical quality of a matching-taper single-cone filling technique following rotary instrumentation compared with lateral compaction after manual preparation: a retrospective study.

OBJECTIVES: The objectives of the study were to evaluate the radiographic technical quality of root canal treatment before and after the implementation of a nickel-titanium rotary (NiTiR) preparation followed by a matching-taper single-cone (mSC) obturation and to detect the procedural errors associated with this technique. MATERIALS AND METHODS: A random sample of 535 patients received root canal treatment at the Department of Conservative Dentistry and Periodontology at the University of Würzburg: 254 teeth were treated in 2002-2003 by using stainless steel instruments (SSI) for preparation and a lateral compaction (LC) technique (classic group (CG)). Two hundred eighty-one teeth were root filled in 2012-2013 employing NiTiR instruments for the root canal shaping and a mSC technique (advanced group (AG)). The quality assessments were based on the radiographic criteria of the European Society of Endodontology. The presence of voids was recorded separately for the apical, central and cervical thirds of the root canals. Procedural errors, such as ledges, apical transportations, perforations and fractured instruments, were detected. The root canal fillings in the CG and AG were compared using chi-squared and Fisher's exact tests. Multivariable logistic regression was performed to investigate the association between the independent variables (patient age, tooth type and type of treatment) and the dependent variables (density and length). RESULTS: Adequate length was achieved significantly more often in the AG compared to the CG for molars (p = 0.017), mandibular teeth (p = 0.013) and primary root canal treatments (p = 0.024). No significant difference was detected between the AG and CG regarding adequate length in general (p = 0.051) or adequate overall quality of root canal filling (p = 0.1). In the AG, a significant decrease in procedural errors was evident (p = 0.019) and decreases in the densities of the root canal fillings in the cervical (p = 0.01) and central (p = 0.01) thirds of the root canals were also observed. Moreover, root canals in elderly patients exhibited fewer voids (p = 0.009). CONCLUSIONS: Rotary root canal preparation followed by a matching-taper single-cone filling technique provides a reliable shaping of the root canal, with fewer procedural errors and a more acceptable filling quality in terms of length and homogeneity in the apical third. Less favourable results were achieved in the central and cervical parts of the root canals. CLINICAL RELEVANCE: The matching-taper single-cone technique seems to effectively obturate well-tapered root canals after adequate rotary instrumentation. Irregularly shaped canals require additional lateral or warm vertical condensation to avoid voids.

A transgenic zebrafish model for monitoring glucocorticoid receptor activity.

Gene regulation resulting from glucocorticoid receptor and glucocorticoid response element interactions is a hallmark feature of stress response signaling. Imbalanced glucocorticoid production and glucocorticoid receptor activity have been linked to socioeconomically crippling neuropsychiatric disorders, and accordingly there is a need to develop in vivo models to help understand disease progression and management. Therefore, we developed the transgenic SR4G zebrafish reporter line with six glucocorticoid response elements used to promote expression of a short half-life green fluorescent protein following glucocorticoid receptor activation. Herein, we document the ability of this reporter line to respond to both chronic and acute exogenous glucocorticoid treatment. The green fluorescent protein expression in response to transgene activation was high in a variety of tissues including the brain, and provided single-cell resolution in the effected regions. The specificity of these responses is demonstrated using the partial agonist mifepristone and mutation of the glucocorticoid receptor. Importantly, the reporter line also modeled the temporal dynamics of endogenous stress response signaling, including the increased production of the glucocorticoid cortisol following hyperosmotic stress and the fluctuations of basal cortisol concentrations with the circadian rhythm. Taken together, these results characterize our newly developed reporter line for elucidating environmental or genetic modifiers of stress response signaling, which may provide insights to the neuronal mechanisms underlying neuropsychiatric disorders such as major depressive disorder.

Neuroprotection by oxygen in acute transient focal cerebral ischemia is dose dependent and shows superiority of hyperbaric oxygenation.

The neuroprotective effect of oxygen after acute stroke in rats has been shown previously. However, the question of optimal dosing still remains unanswered. Thus, we investigated the use of oxygen at different concentrations by either normobaric oxygenation (NBO) or hyperbaric oxygenation (HBO) at different pressures in a model of transient ischemia/reperfusion in rats. Animals underwent 90 min of middle cerebral artery occlusion (MCAO) followed by 90 min of reperfusion before oxygen treatment. Oxygen was applied either by NBO (100% O(2); 1.0 absolute atmosphere, ATA) or HBO (100% O(2); 1.5, 2.0, 2.5 or 3.0 ATA) for 1 h. Primary endpoints were infarct volume and clinical outcome measured 24 h and 7 days following the MCAO. A statistically significant and long-lasting reduction in infarct volume was seen in the HBO 2.5 ATA and 3.0 ATA groups over a period of 7 days. The reduced infarct volume was accompanied with a statistically significant improvement in clinical outcome in the high-dose oxygen-treated groups. The presented data indicate that oxygen is a highly neuroprotective molecule in transient focal cerebral ischemia in rats, when applied early and at high doses. The effect is dose dependent and shows a superiority of HBO over NBO, when the primary endpoints infarct volume reduction and clinical outcome are analyzed. These data are important for the development of new acute stroke treatment studies in humans.

Assessment of trabecular bone structure comparing magnetic resonance imaging at 3 Tesla with high-resolution peripheral quantitative computed tomography ex vivo and in vivo.

UNLABELLED: In vivo high-resolution peripheral quantitative micro-CT (HR-pQCT) is a new modality for imaging peripheral sites like the distal tibia and the distal radius, providing structural bone parameters. Comparing HR-pQCT with MRI, we found that both modalities are capable of offering meaningful information on trabecular structure. BACKGROUND: Magnetic resonance imaging (MRI) has emerged as the leading in vivo method for measuring trabecular bone micro-architecture and providing structural information. Recently, an in vivo HR-pQCT modality was introduced for imaging peripheral sites like the distal tibia and the distal radius, providing structural bone parameters. The goal of this work was to compare and evaluate the performances and in vivo capabilities of HR-pQCT in comparison with MRI at 3 Tesla. METHODS: To this end images of 8 human specimens (5 tibiae and 3 radii) and 11 participants (6 tibia and 5 radii) were acquired with both modalities. Additionally, the radius specimens were scanned with micro-CT (muCT), which was used as a standard of reference. Structural parameters calculated from MRI were compared with results from HR-pQCT images and additionally muCT for the radii specimens. RESULTS: High correlations (r > 0.7) were found for trabecular number and trabecular spacing between the two modalities in vivo and ex vivo. 2D and 3D analysis revealed high correlations (r > 0.8) in structural bone parameters for all measurements. Using micro-CT as standard of reference both results from QCT and MRI correlated well. CONCLUSION: Both imaging modalities were found to perform equally well regarding trabecular bone measurements.

[Compliance of the normal-sized aorta in adolescents with Marfan syndrome: comparison of MR measurements of aortic distensibility and pulse wave velocity]. / Compliance der morphologisch unauffälligen Aorta bei Jugendlichen mit Marfan Syndrom: Vergleich von MR-Messungen der aortalen Dehnbarkeit und der Pulswellengeschwindigkeit.

PURPOSE: To compare the aortic compliance of the normal-sized aorta of adolescents with Marfan syndrome and healthy controls using MR measurements of the aortic distensibility and pulse wave velocity. MATERIALS AND METHODS: Fourteen patients (median age: 15 years) and 11 healthy subjects (23 [12 - 32] years) were examined at 1.5 T. The MR protocol included 2D steady-state free precession (SSFP)-CINE MRI of the aortic distensibility and PC-MRI of the pulse wave velocity. All measurements were positioned perpendicular to the descending aorta at the level of the diaphragm for assessing the changes in the aortic cross-sectional areas and additionally above and below this plane for assessing the pulse wave velocity. In addition contrast-enhanced 3D-MR angiography was performed in adolescents with Marfan syndrome to exclude morphologic changes and to prove normal-sized aorta. RESULTS: Compared with control subjects, adolescents with Marfan syndrome had significantly decreased distensibility and significantly increased pulse wave velocity (chi (2)-test, p = 0.0002) using an age-related non-linear regression analysis. The related aortic compliance was significantly decreased (chi (2)-test, p = 0.0002). There was a good correlation between the two methods (r = 0.86). A low intraobserver variability was found for both methods (< or = 2 %). CONCLUSIONS: MRI allows detecting abnormal elastic aortic wall properties already in the normal-sized aorta of adolescents with Marfan syndrome. Monitoring of these properties could be relevant for evaluating disease onset and progression. MRI has the potential value of compliance measurements for the follow-up and to guide therapy indications.

Fully balanced steady-state 3D-spin-echo (bSSSE) imaging at 3 Tesla.

In this work a 3D fully balanced steady-state spin-echo sequence (bSSSE) was designed and implemented on a 3-Tesla whole-body MR scanner. In contrast to previously introduced nonbalanced SSSE (nbSSSE) sequences, bSSSE features fully rewound imaging gradients. Additionally, all phase encodings and the readout prephasing are performed after the refocusing pulse. Its performance was compared with previously used gradient-echo (GE) and nbSSSE sequences. The signal response of each sequence was simulated by numerical solution of the Bloch equation. All sequences were then optimized based on these simulations for high-resolution (HR) imaging of tissues with short relaxation times and applied to in vivo HR-MRI of trabecular bone. bSSSE outperformed nbSSSE sequences in terms of SNR and SNR efficiency. Measurements of structural bone parameters revealed thinner trabeculae and a lower bone/bone-marrow fraction for the bSSSE sequence compared to the nbSSSE sequence. By applying a parallel imaging technique based on generalized autocalibrating partially parallel acquisition (GRAPPA) with a reduction factor of 2, we were able to maintain a clinically feasible scan time using bSSSE. Comparisons of structural bone parameters revealed a difference of less than 3% for all structural parameters between parallel and conventional imaging.

Wavelet Based Characterization of Vertebral Trabecular Bone Structure from MR Images of Specimen at 3 Tesla Compared to MicroCT Measurements.

Trabecular bone structure and bone density contribute to the strength of bone and are important in the study of osteoporosis. Wavelets are a powerful tool to characterize and quantify texture in an image. In this study the thickness of trabecular bone was analyzed in 8 cylindrical cores of the vertebral spine. Images were obtained from 3 Tesla (T) magnetic resonance imaging (MRI) and micro-computed tomography (μCT). Results from the wavelet based analysis of trabecular bone were compared with standard two-dimensional (2D) structural parameters (analogous to bone histomorphometry) obtained using mean intercept length (MR images) and direct three-dimensional (3D) distance transformation methods (μCT images). Additionally, the bone volume fraction was determined from MR images. We conclude that the wavelet based analyses delivers comparable results to the established MR histomorphometric measurements. The average deviation in trabecular thickness was less than one pixel size between the wavelet and the standard approach for both MR and μCT analysis. Since the wavelet based method is less sensitive to image noise, we see an advantage of wavelet analysis of trabecular bone for MR imaging when going to higher resolution.

[The biofeedback sole tested is a suitable device for avoiding overload under partial load]. / Kontrollierte Teilbelastung unter Verwendung einer Biofeedbackschuhsohle.

During healing of fractures and after implantation of artificial joints mobilization with partial load is a widespread and well-accepted therapeutic principle. Translating our recommendations into action, however, overtaxes many of the patients. The use of the biofeedback sole tested should guarantee avoiding overload under partial weight-bearing conditions. The distribution of the strain sensors in the biofeedback sole was primarily checked with the help of an established system controlling the distribution of strain (PAROTEC) in 17 voluntary, healthy people. Afterwards the biofeedback sole was connected to a digital recording system to analyze the number of steps overloaded in different test cycles in 50 volunteers while using crutches and performing partial load with 20 kg. The distribution of the strain sensors in the biofeedback sole is correct to control partial load under weight bearing with 20 kg. The permanent use of the biofeedback sole showed highly significant advantages in avoiding overload. The number of steps overloaded was reduced by 77%. This effect is independent from body weight or age.

Efficient translation of mRNAs in influenza A virus-infected cells is independent of the viral 5' untranslated region.

We test the hypothesis that the translation machinery in cells infected by influenza A virus efficiently translates only mRNAs that possess the influenza viral 5' untranslated region (5'-UTR) by introducing mRNAs directly into the cytoplasm of infected cells. This strategy avoids effects due to the inhibition of the nuclear export of cellular mRNAs mediated by the viral NS1 protein. In one approach, we transfect in vitro synthesized mRNAs into infected cells and demonstrate that these mRNAs are efficiently translated whether or not they possess the influenza viral 5'-UTR. In the second approach, an mRNA is synthesized endogenously in the cytoplasm of influenza A virus infected cells by a constitutively expressed T7 RNA polymerase. Although this mRNA is uncapped and lacks the influenza viral 5'-UTR sequence, it is efficiently translated in infected cells via an internal ribosome entry site. We conclude that the translation machinery in influenza A virus infected cells is capable of efficiently translating all mRNAs and that the switch from cellular to virus-specific protein synthesis that occurs during infection results from other processes.

Converting from general anesthesia to cervical block anesthesia for carotid endarterectomy.

Between January 1, 1992 and June 30, 1998, vascular and general surgery residents performed 401 carotid endarterectomies (185 cervical block, 216 general anesthesia) under supervision of vascular attending surgeons. In January 1995, cervical block anesthesia was newly instituted. Initially anesthesiologists were randomly assigned to these cases and anesthetic technique was not standardized. At the surgeons' insistence later in the series, three specially trained anesthesiologists routinely administered cervical block anesthesia. As experience grew, surgeons realized that operating time greater than 2 hours and high neck dissections requiring mandibular retraction were poorly tolerated by cervical block anesthesia patients but that repeat carotid endarterectomies could be safely performed. Shunts were selectively inserted if significant electroencephalographic changes occurred or carotid stump pressures were less than 50 mm Hg systolic when general anesthesia was used; neurologic changes occurred when cervical block anesthesia was used; or there was a history of ipsilateral stroke for either anesthetic method. Despite an initial learning curve with cervical block anesthesia, the stroke-mortality rate was similar between the two groups (general anesthesia: 1.9% [four cerebrovascular accidents, two deaths]; cervical block anesthesia: 1.6% [two cerebrovascular accidents, two deaths]). Eight (12%) of the first 66 cervical block anesthesia patients were converted to general anesthesia compared to 2 (1.7%) of the most recent 119 patients with cervical block anesthesia (p = 0.03). After cervical block anesthesia was initiated, seven of the first eight repeat carotid endarterectomies were performed using general anesthesia compared to one of the most recent six repeat cases (p = 0.02). As surgeons' comfort with cervical block anesthesia increased, 94% (100) of the most recent consecutive 106 carotid endarterectomies were performed using this technique. These results suggest that carotid endarterectomy can be performed as safely using cervical block anesthesia as general anesthesia, specialized anesthesiologists are essential to achieve a successful outcome, selected repeat carotid endarterectomies can be performed using cervical block anesthesia, very cephalad lesions are best treated using general anesthesia, and surgical trainees can safely perform carotid endarterectomy under attending surgeon supervision if the operation is carried out expeditiously.

Should duplex ultrasonography be performed for surveillance of femoropopliteal and femorotibial arterial prosthetic bypasses?

Duplex ultrasonography (DU) has been shown to be beneficial for surveillance of lower extremity vein bypasses. However, DU as part of surveillance program for prosthetic grafts is not widely accepted. The purpose of this report was to determine if DU could reliably detect failing prosthetic infrainguinal arterial bypasses and if there were differences in predictability between femoropopliteal (FP) and femorotibial (FT) prosthetic grafts. Between January 1992 and December 1997, 89 infrainguinal grafts in 66 patients were entered into our postoperative prosthetic graft surveillance protocol, which included clinical evaluation, segmental pressures, pulse volume recordings, and DU performed every 3 months. Patients with follow-up of less than 3 months were excluded unless the graft thrombosed. An abnormal DU considered predictive of graft failure included (1) peak systolic velocity (PSV) > 300 cms/sec at inflow or outflow arteries, in the graft or at an anastomosis (unless an adjunctive arteriovenous fistula had been performed); (2) adjacent PSV ratio > 3.0; (3) uniform PSVs < 45 cms/sec; or (4) monophasic signals throughout the graft. DU was considered to have correctly diagnosed a failing graft if a stenosis > 75% the luminal diameter of the graft, at an anastomosis, or in an inflow/outflow artery was confirmed by operative or arteriographic findings or if the graft thrombosed after an abnormal DU but before intervention. Our results support the routine use of DU as a part of a graft surveillance protocol for femorotibial, but not femoropopliteal, prosthetic grafts.

The 3'-end-processing factor CPSF is required for the splicing of single-intron pre-mRNAs in vivo.

We describe a new approach to elucidate the role of 3'-end processing in pre-mRNA splicing in vivo using the influenza virus NS1A protein. The effector domain of the NS1A protein, which inhibits the function of the CPSF and PABII factors of the cellular 3'-end-processing machinery, is sufficient for the inhibition of not only 3'-end formation but also the splicing of single-intron pre-mRNAs in vivo. We demonstrate that inhibition of the splicing of single-intron pre-mRNAs results from inhibition of 3'-end processing, thereby establishing that 3'-end processing is required for the splicing of a 3' terminal intron in vivo. Because the NS1A protein causes a global suppression of 3'-end processing in trans, we avoid the ambiguities caused by the activation of cryptic poly(A) sites that occurs when mutations are introduced into the AAUAAA sequence in the pre-mRNA. In addition, this strategy enabled us to establish that the function of a particular 3'-end-processing factor, namely CPSF, is required for the splicing of single-intron pre-mRNAs in vivo: splicing is inhibited only when the effector domain of the NS1A protein binds and inhibits the function of the 30-kDa CPSF protein in 3'-end formation. In contrast, the 3'-end processing factor PABII is not required for splicing. We discuss the implications of these results for cellular and influenza viral mRNA splicing.

The active sites of the influenza cap-dependent endonuclease are on different polymerase subunits.

The cap-dependent endonuclease of the influenza viral RNA polymerase, which produces the capped RNA primers that initiate viral mRNA synthesis, is comprised of two active sites, one for cap binding and one for endonuclease cleavage. We identify the amino acid sequences that constitute these two active sites and demonstrate that they are located on different polymerase subunits. Binding of the 5' terminal sequence of virion RNA (vRNA) to the polymerase activates a tryptophan-rich, cap-binding sequence on the PB2 subunit. At least one of the tryptophans functions in cap binding, indicating that this active site is probably similar to that of other known cap-binding proteins. Endonuclease cleavage, which is activated by the subsequent binding of the 3' terminal sequence of vRNA, resides in a PB1 sequence that contains three essential acidic amino acids, similar to the active sites of other enzymes that cut polynucleotides to produce 3'-OH ends. These results, coupled with those of our previous study, provide a molecular map of the five known essential active sites of the influenza viral polymerase.

Influenza B virus NS1 protein inhibits conjugation of the interferon (IFN)-induced ubiquitin-like ISG15 protein.

Of the several hundred proteins induced by interferon (IFN) alpha/beta, the ubiquitin-like ISG15 protein is one of the most predominant. We demonstrate the novel way in which the function of the ISG15 protein is inhibited by influenza B virus, which strongly induces the ISG15 protein: a specific region of the influenza B virus NS1 protein, which includes part of its effector domain, blocks the covalent linkage of ISG15 to its target proteins both in vitro and in infected cells. We identify UBE1L as the E1 enzyme that catalyzes the first activation step in the conjugation of ISG15, and show that the NS1B protein inhibits this activation step in vitro. Influenza A virus employs a different strategy: its NS1 protein does not bind the ISG15 protein, but little or no ISG15 protein is produced during infection. We discuss the likely basis for these different strategies.