Purification of coronavirus virions for Cryo-EM and proteomic analysis

Purification of intact enveloped virus particles can be useful as a first step in understanding the structure and function of both viral and host proteins that are incorporated into the virion. Purified preparations of virions can be used to address these questions using techniques such as mass spectrometry proteomics. Recent studies on the proteome of coronavirus virions have shown that in addition to the structural proteins, accessory and non-structural virus proteins and a wide variety of host cell proteins associate with virus particles. To further study the presence of virion proteins, high quality sample preparation is crucial to ensure reproducible analysis by the wide variety of methods available for proteomic analysis.

Assessing the carcinogenic potential of low-dose exposures to chemical mixtures in the environment: the challenge ahead

Lifestyle factors are responsible for a considerable portion of cancer incidence worldwide, but credible estimates from the World Health Organization and the International Agency for Research on Cancer (IARC) suggest that the fraction of cancers attributable to toxic environmental exposures is between 7% and 19%. To explore the hypothesis that low-dose exposures to mixtures of chemicals in the environment may be combining to contribute to environmental carcinogenesis, we reviewed 11 hallmark phenotypes of cancer, multiple priority target sites for disruption in each area and prototypical chemical disruptors for all targets, this included dose-response characterizations, evidence of low-dose effects and cross-hallmark effects for all targets and chemicals. In total, 85 examples of chemicals were reviewed for actions on key pathways/mechanisms related to carcinogenesis. Only 15% (13/85) were found to have evidence of a dose-response threshold, whereas 59% (50/85) exerted low-dose effects. No dose-response information was found for the remaining 26% (22/85). Our analysis suggests that the cumulative effects of individual (non-carcinogenic) chemicals acting on different pathways, and a variety of related systems, organs, tissues and cells could plausibly conspire to produce carcinogenic synergies. Additional basic research on carcinogenesis and research focused on low-dose effects of chemical mixtures needs to be rigorously pursued before the merits of this hypothesis can be further advanced. However, the structure of the World Health Organization International Programme on Chemical Safety 'Mode of Action' framework should be revisited as it has inherent weaknesses that are not fully aligned with our current understanding of cancer biology.

Challenges in quantifying wind generation's contribution to securing peak demand

Wind generation's contribution to supporting peak electricity demand is one of the key questions in wind integration studies. Differently from conventional units, the available outputs of different wind farms cannot be approximated as being statistically independent, and hence near-zero wind output is possible across an entire power system. This paper will review the risk model structures currently used to assess wind's capacity value, along with discussion of the resulting data requirements. A central theme is the benefits from performing statistical estimation of the joint distribution for demand and available wind capacity, focusing attention on uncertainties due to limited histories of wind and demand data; examination of Great Britain data from the last 25 years shows that the data requirements are greater than generally thought. A discussion is therefore presented into how analysis of the types of weather system which have historically driven extreme electricity demands can help to deliver robust insights into wind's contribution to supporting demand, even in the face of such data limitations. The role of the form of the probability distribution for available conventional capacity in driving wind capacity credit results is also discussed.

The proteome of the infectious bronchitis virus Beau-R Virion

Infectious bronchitis is a highly contagious respiratory disease of poultry caused by the coronavirus IBV. It was thought that coronavirus virions were composed of three major viral structural proteins, until investigations of other coronaviruses showed that coronavirus virions also include viral non-structural and group specific proteins as well as host cell proteins. To study the proteome of IBV virions, virus was grown in embryonated chicken eggs and purified by sucrose gradient ultracentrifugation and analysed by mass spectrometry proteomic. Analysis of three preparations of purified IBV yielded the three expected structural proteins plus thirty-five additional virion-associated host proteins. Virion-associated host proteins had a diverse range of functional attributions, being involved in cytoskeleton formation, RNA binding and protein folding pathways. Some of these proteins were unique to this study, whilst others were found to be orthologous to proteins identified in SARS-CoV virions, and also virions from a number of other RNA and DNA viruses. Together these results demonstrate that coronaviruses have the capacity to incorporate a substantial variety of host protein, which may have implications for the disease process.

Ex Vivo Accuracy of an Apex Locator Using Digital Signal Processing in Primary Teeth

The purpose of this study was to evaluate ex vivo the accuracy an electronic apex locator during root canal length determination in primary molars. Methods: One calibrated examiner determined the root canal length in 15 primary molars (total=34 root canals) with different stages of root resorption. Root canal length was measured both visually, with the placement of a K-file 1 mm short of the apical foramen or the apical resorption bevel, and electronically using an electronic apex locator (Digital Signal Processing). Data were analyzed statistically using the intraclass correlation (ICC) test. Results: Comparing the actual and electronic root canal length measurements in the primary teeth showed a high correlation (ICC=0.95) Conclusions: The Digital Signal Processing apex locator is useful and accurate for apex foramen location during root canal length measurement in primary molars. (Pediatr Dent 200937:320-2) Received April 75, 2008 vertical bar Lost Revision August 21, 2008 vertical bar Revision Accepted August 22, 2008

Unilateral Correction of Space Loss in the Mixed Dentition

Distalization of maxillary molars is indicated for correction of Class II dental malocclusion and for space gain in cases of space deficiency. The ideal treatment with an intraoral fixed appliance for molar distalization should fulfill the following requirements: patient compliance; acceptable esthetics; comfort; minimum anterior anchor loss (as evidenced by inclination of incisors); bodily movement of the molars to avoid undesirable effects and unstable outcomes; and minimum time required during sessions for placement and activations. The purpose of this paper was to present an alternative treatment for space recovery in the area of the maxillary right second premolar when there has been significant mesial movement of the permanent maxillary right first molar. We used a modified appliance that allows unilateral molar distalization in cases of unilateral tooth/arch size discrepancy using the opposite side as anchor, thus reducing the mesialization of the anterior teeth. (Pediatr Dent 2008;30:334-41) Received August 17, 2006 / Last Revision October 17, 2007 / Revision Accepted October 17, 2007

Efficacy of Microwaves and Chlorhexidine on the Disinfection of Pacifiers and Toothbrushes: An In Vitro Study

Purpose: The purpose of this study was to evaluate, in vitro, the contamination of toothbrushes and pacifiers by Streptococcus mutans, and the efficacy of microwave and chlorhexidine for their disinfection. Methods: Sixty pacifiers and 60 toothbrushes were contaminated with S mutans and then divided into groups according to the disinfection protocol: Group 1-chlorhexidine solution; Group 2-microwave sterilization; and Group 3-sterile tap water. The devices were evaluated microbiologically as to the formation of S mutans colonies/biofilms and were examined by scanning electron microscopy. The results were submitted for statistical analysis by Friedman`s test at a 5% significance level. Results: The results of both types of evaluation showed a large number of S mutans colonies/biofilms after spraying with sterile tap water, and chlorhexidine spraying and microwaving were effective in eliminate colonies/biofilms. Groups 1 and 2 were statistically similar to each other (P>.05) and differed significantly from Group 3 (P<.05). Conclusions: The 0.12% chlorhexidine solution spray and 7 minutes of microwave irradiation were effective for disinfection of pacifiers and toothbrushes. (Pediatr Dent 2011;33:10-3) Received July 29, 2009 I Last Revision January 26, 2010 I Accepted March 10, 2010

A Prospective Follow-up Study of 44 Mandibular Immediately Loaded Implants Using Resonance Frequency Analysis: Preliminary 1-Year Results

Purpose: This clinical study aimed to evaluate initial, 4-months, and 1-year stability of immediately loaded dental implants inserted according to a protocol of lower rehabilitation with prefabricated bars. Materials and Methods: The sample was composed of 11 edentulous patients. In each patient, 4 interforaminal implants were inserted. Immediately after implant installation, resonance frequency analysis (RFA) for each fixation was registered as well as after 4 months and 1 year with the prosthetic bar removed as it is a screwed system. Results: The clinical implant survival rate was 100%. The RFA showed an increase in stability after 4 months from 64.09 +/- 648 to 64.31 +/- 4.96 and I year, 67.11 +/- 4.37. The analysis of variance showed a statistically significant result (P = 0.015) among implant stability quotient values for the different periods evaluated. Tukey test results showed statistically significant differences between 1-year results and the initial periods but there was no statistically significant difference between initial and 4-month results (P > 0.05). Conclusion: These preliminary 1-year results indicate that immediate loading of mandibular dental implants using the studied prefabricated bars protocol is a reliable treatment as it is in accordance with the results described in the literature for other similar techniques. (Implant Dent 2009; 18:530-538)

COLOR STABILITY OF DENTAL CERAMICS SUBMITTED TO ARTIFICIAL ACCELERATED AGING AFTER REPEATED FIRINGS

Statement of problem. Color stability is an important factor to ensure the long-term clinical success of ceramic restorations. There is a lack of information on how color is affected by fabrication procedures, such as the number of firings. Purpose. The purpose of this study was to evaluate the effects that the number of firings and type of substrate have on the color stability of dental ceramic submitted to artificial accelerated aging. Material and methods. Sixty specimens were fabricated: 30 metal ceramic (Verabond II + IPS d.SIGN) and 30 all-ceramic (IPS d.SIGN). Specimens were divided into 3 groups (n=10), and submitted to 2, 3, or 4 firings (+/- 900 degrees C), respectively, according to the manufacturer`s instructions. Color readings were obtained with a spectro photometer before and after artificial accelerated aging, and L*, a*, and b* coordinates and total color variation (Delta E) were analyzed (2-way ANOVA, Bonferroni, (alpha=05). Results. For metal ceramic specimens, differences for the L* coordinates were significant (P<.05) only for the group submitted to 3 firings. With respect to the all-ceramic specimens, smaller L* coordinates were obtained for greater a* and b* coordinates, indicating that the greater the number of firings, the darker and more reddish/yellowish the specimen. All Delta E values, for all groups, were below 1.0. All-ceramic specimens submitted to 3 and 4 firings presented Delta E means differing statistically (P<.05) from those of the metal ceramic group. Conclusions. The type of substrate and number of firings affected the color stability of the ceramic material tested. Artificial accelerated aging did not produce perceptible color stability changes (Delta E<1.0). (J Prosthet Dent 2009-101:13-18)

Bond Strength and Etching Pattern of Adhesive Systems to Enamel: Effects of Conditioning Time and Enamel Preparation

Statement of the problem: The performance of self-etch systems on enamel is controversial and seems to be dependent on the application technique and the enamel preparation. Purpose of the Study: To examine the effects of conditioning time and enamel surface preparation on bond strength and etching pattern of adhesive systems to enamel. Materials and Methods: Ninety-six teeth were divided into 16 conditions (N = 6) in function of enamel preparation and conditioning time for bond strength test. The adhesive systems OptiBond FL (Kerr, Orange, CA, USA), OptiBond SOLO Plus (Kerr), Clearfil SE Bond (Kuraray, Osaka, Japan), and Adper Prompt L-Pop (3M ESPE, St. Paul, MN, USA) were applied on unground or ground enamel following the manufacturers` directions or doubling the conditioning time. Cylinders of Filtek Flow (0.5-mm height) were applied to each bonded enamel surface using a Tygon tube (0.7 mm in diameter; Saint-Gobain Corp., Aurora, OH, USA). After storage (24 h/37 degrees C), the specimens were subjected to shear force (0.5 mm/min). The data were treated by a three-way analysis of variance and Tukey`s test (alpha = 0.05). The failure modes of the debonded interfaces and the etching pattern of adhesives were observed using scanning electron microscopy. Results: Only the main factor ""adhesive"" was statistically significant (p < 0.001). The lowest bond strength value was observed for OptiBond FL. The most defined etching pattern was observed for 35% phosphoric acid and for Adper Prompt L-Pop. Mixed failures were observed for all adhesives, but OptiBond FL showed cohesive failures in resin predominantly. Conclusions: The increase in the conditioning time as well as the enamel pretreatment did not provide an increase in the resin-enamel bond strength values for the studied adhesives. CLINICAL SIGNIFICANCE The surface enamel preparation and the conditioning time do not affect the performance of self-etch systems to enamel. (J Esthet Restor Dent 20:322-336, 2008)

Effect of steeping conditions (sulfur dioxide, lactic acid, and temperature) on starch yield, starch quality, and germ quality from the Intermittent Milling and Dynamic Steeping process (IMDS) for a Brazilian corn hybrid

Effect of lactic acid, SO2, temperature, and their interactions were assessed on the dynamic steeping of a Brazilian dent corn (hybrid XL 606) to determine the ideal relationship among these variables to improve the wet-milling process for starch and corn by-products production. A 2x2x3 factorial experimental design was used with SO2 levels of 0.05 and 0.1% (w/v), lactic acid levels of 0 and 0.5% (v/v), and temperatures of 52, 60, and 68degreesC. Starch yield was used as deciding factor to choose the best treatment. Lactic acid added in the steep solution improved the starch yield by an average of 5.6 percentage points. SO2 was more available to break down the structural protein network at 0.1% than at the 0.05% level. Starch-gluten separation was difficult at 68degreesC. The lactic acid and SO2 concentrations and steeping temperatures for better starch recovery were 0.5, 0.1, and 52degreesC, respectively. The Intermittent Milling and Dynamic Steeping (IMDS) process produced, on average, 1.4% more starch than the conventional 36- hr steeping process. Protein in starch, oil content in germ, and germ damage were used as quality factors. Total steep time can be reduced from 36 hr for conventional wet-milling to 8 hr for the IMDS process.

Contribuições para verificação automática de applets javacard

The widespread growth in the use of smart cards (by banks, transport services, and cell phones, etc) has brought an important fact that must be addressed: the need of tools that can be used to verify such cards, so to guarantee the correctness of their software. As the vast majority of cards that are being developed nowadays use the JavaCard technology as they software layer, the use of the Java Modeling Language (JML) to specify their programs appear as a natural solution. JML is a formal language tailored to Java. It has been inspired by methodologies from Larch and Eiffel, and has been widely adopted as the de facto language when dealing with specification of any Java related program. Various tools that make use of JML have already been developed, covering a wide range of functionalities, such as run time and static checking. But the tools existent so far for static checking are not fully automated, and, those that are, do not offer an adequate level of soundness and completeness. Our objective is to contribute to a series of techniques, that can be used to accomplish a fully automated and confident verification of JavaCard applets. In this work we present the first steps to this. With the use of a software platform comprised by Krakatoa, Why and haRVey, we developed a set of techniques to reduce the size of the theory necessary to verify the specifications. Such techniques have yielded very good results, with gains of almost 100% in all tested cases, and has proved as a valuable technique to be used, not only in this, but in most real world problems related to automatic verification

Finite Element Modeling for Development and Optimization of a Bone Plate for Mandibular Fracture in Dogs

This study aimed to develop a plate to treat fractures of the mandibular body in dogs and to validate the project using finite elements and biomechanical essays. Mandible prototypes were produced with 10 oblique ventrorostral fractures (favorable) and 10 oblique ventrocaudal fractures (unfavorable). Three groups were established for each fracture type. Osteosynthesis with a pure titanium plate of double-arch geometry and blocked monocortical screws offree angulanon were used. The mechanical resistance of the prototype with unfavorable fracture was lower than that of the fcworable fracture. In both fractures, the deflection increased and the relative stiffness decreased proportionally to the diminishing screw number The finite element analysis validated this plate study, since the maximum tension concentration observed on the plate was lower than the resistance limit tension admitted by the titanium. In conclusion, the double-arch geometry plate fixed with blocked monocortical screws has sufficient resistance to stabilize oblique,fractures, without compromising mandibular dental or neurovascular structures. J Vet Dent 24 (7); 212 - 221, 2010

Bone Spreading and Standardized Dilation of Horizontally Resorbed Bone: Technical Considerations

The uses of a new bone spreading technique with simultaneous implant placement are discussed. The spreading system is an alternative technique to Summers' osteotome. Specific screw designs (spreader) served to laterally compress the bone to increase the cancellous density adjacent to the site. The spreader achieved a controlled and standardized dilation of horizontal bone. The advantages, material selection, and the application of this new procedure are detailed. (Implant Dent 2009; 18:119-125)