Debonding of Leucite-reinforced Glass-ceramic Veneers Using Er, Cr:YSGG Laser Device: Optimizing Speed with Thermal Safety.

CLINICAL RELEVANCE: Removing laminate veneers on anterior teeth by using an Er,Cr:YSGG dental laser can be completed faster than previously reported while maintaining thermal safety.

[Efficacy and safety of neoadjuvant apatinib in combination with dose-dense paclitaxel and carboplatin in locally advanced triple negative breast cancer patients].

Objective: To observe the short-term efficacy and safety of apatinib in combination with dose-dense paclitaxel and carboplatin in locally advanced triple-negative breast cancer (TNBC) patients. Methods: From September 2018 to September 2019, 17 stage Ⅱ/Ⅲ TNBC patients were enrolled in this single arm, single center prospective phase Ⅱ study. They received neoadjuvant treatment of apatinib 250 mg per day, paclitaxel 175 mg/m(2) on 1(st) day and a dose of carboplatin according to the area under curve (AUC)=4 on 2(nd) day, every 14 days as a cycle. Results: By January 2020, 16 cases completed 4-7 cycles of apatinib treatment and 4-8 cycles of chemotherapy. The median cycles of apatinib treatment and chemotherapy were 5 cycles and 6 cycles, respectively. Two cases achieved complete responses (CR), 12 achieved partial responses (PR), 2 achieved stable diseases (SD) and no progressive disease was observed. The objective response rate (ORR) was 87.5%, disease control rate (DCR) was 100%. By January 2020, among 12 patients who received surgery, 8 achieved pathologic complete response (pCR, 66.7%). The grade Ⅲ/Ⅳ adverse events included: neutropenia, thrombocytopenia in 3 cases (18.8%) each, anemia, fatigue, arrhythmia and alanine aminotransferase (ALT) elevation in 1 case each. Apatinib was interrupted in 5 cases, and was discontinued in 3 cases; chemotherapy dosage was reduced in 1 case. Conclusion: Apatinib in combination with dose-dense paclitaxel and carboplatin neoadjuvant therapy are effective and well tolerated in locally advanced TNBC patients.

Stress analysis during jaw movement based on vivo computed tomography images from patients with temporomandibular disorders.

The purpose of this study is to develop three-dimensional (3D) finite element models of temporomandibular joints (TMJs) and to investigate stress distributions. To determine the causes of temporomandibular disorders (TMDs), the magnitude and location of the maximum stresses under physiological loading must be considered. Stress analysis TMD models were reconstructed from computed tomography (CT) data. Several studies have investigated finite element TMJ models, but few have used a bilateral mandible model that includes jaw closing and maximum opening. In this study, the authors defined an asymmetry index for the different stress values on each side joint; this index has not yet been investigated. According to clinical observation, one joint affects the other side joint during mastication. Three symptom-free volunteers and three symptomatic patients were selected as the control group (CG) and TMD group (TG), respectively. For the TG, data analysis indicated that the condyle was asymmetrical during jaw closing, while both the condyle and disc were slightly asymmetrical during jaw opening. The maximum stresses did not significantly differ between the CG and TG for either closing or opening of the jaw. The results of this study have a potential clinical benefit in terms of proving superior biomechanical behaviour.

Botryomycosis presenting as nasal cutaneous fistulas caused by Prevotella melaninogenica.

Botryomycosis is an uncommon chronic suppurative granulomatous bacterial infection that can affect the skin and viscera. Clinically, lesions typically consist of small tender nodules from which draining sinuses may develop to expel a purulent discharge. Histopathological features include characteristic aggregation of microorganisms (grain) within the inflammatory infiltrate. The commonest causative organisms are Staphylococcus aureus and Pseudomonas aeruginosa, of others. Botryomycosis resulting from Prevotella melaninogenica has not been reported previously. We report the case of a middle-aged patient with botryomycosis presenting as nasal cutaneous fistulas caused by P. melaninogenica, which was successfully treated with surgical intervention combined with systemic antibiotic treatment.

Effect of damping properties on fracture resistance of root filled premolar teeth: a dynamic finite element analysis.

AIM: To evaluate the ex vivo effects of damping on stress concentration in root filled premolar teeth. METHODOLOGY: Damping ratios of maxillary premolar teeth that had undergone root canal treatment were tested in a laboratory model. In addition, two-dimensional finite element (FE) models were established for dynamic analysis. RESULTS: The mean-damping ratio was significantly lower in premolar teeth that had undergone root canal preparation (8.50 +/- 0.53%) than in unprepared teeth (14.42 +/- 2.17%) (P < 0.05). However, root filling had a significant positive effect on the damping ratio of the tooth (10.84 +/- 1.70%) (P < 0.05). When the damping ratio was taken into consideration, FE analysis revealed that peak stresses in the apical one-third of the root on the buccal side were reduced by 31.8% when mastication forces were applied on the palatal cusp and occlusal fossa. CONCLUSION: Pulp tissue plays an important role in providing protective effects when teeth are subjected to a dynamic load. However, root filled teeth do not provide such protective effects.

Detection of the furcation involvement of a multi-rooted molar using natural frequency analysis: a numerical approach.

The aim of this study was to evaluate the potential for using the natural frequency (NF) as a parameter to detect vertical bone loss at the furcation of human molars as well as to assess the role that the surrounding bone plays in maintaining molar stability. A three-dimensional finite element model of the human maxillary molar was built. The NF values of the molar modal were calculated with one-sided, two-sided, and three-sided vertical bone loss. It was found that the change in the NF was less than 25 per cent in molars with a one-sided defect when the bone level varied by 10 mm from the cementoenamel junction. However, when a three-sided bony defect was simulated, the change in the NF ranged from 40 to 60 per cent. In addition, it was found that bone loss that had reached the furcation entrance (4 mm) resulted in a sharp change in the NF value. Furthermore, it was found that bone loss involving the mesial and distal surfaces resulted in a larger decrease in the NF value compared with bone loss involving the buccal and palatal surfaces. These results demonstrated that the bone surrounding the mesial and distal sides plays a more important role in maintaining molar stability than does the bone surrounding the buccal and palatal sides.

Natural frequency analysis of tooth stability under various simulated types and degrees of alveolar vertical bone loss.

The aim of this study was to test natural teeth stability under various simulated types and degrees of alveolar vertical bone loss, as well as to assess the role that the surrounding bone played for maintaining tooth stability. A three-dimensional finite element model of the human maxillary central incisor with surrounding tissue, including periodontal ligament, enamel, dentin, pulp, and alveolar bone, was established. One side and multiple vertical bone loss were simulated by means of decreasing the surrounding bone level apically from the cemento-enamel junction in 1 mm steps incrementally downward for 10 mm. Natural frequency values of the incisor model with various types and degrees of bone loss were then calculated. The results showed that, with one-sided bone resorption, the model with labial bone loss had the lowest natural frequency decreasing rates (8.2 per cent). On the other hand, in cases of multiple bone loss, vertical bone resorption at the mesial and distal sides had more negative effects on tooth stability compared to vertical bone losses on facial and lingual sides. These findings suggest that the natural frequency method may be a useful, auxiliary clinical tool for diagnosis of vertical periodontal diseases.

Design of a stability-detecting device for dental implants.

Resonance frequency (RF) analysis technology was used to design a dental implant stability detector. The device uses a miniature-sized electromagnetic triggering rod to elicit vibration in a dental implant. Vibrational signals were recorded via an acoustic receiver. To assess the in vivo performance of the test apparatus, animal models were used. Implants were placed in the left tibia of 12 rabbits using a conventional surgical procedure. Standard 3.2 mm x 8 mm implants were placed in each test tibia with pre-tapping cavities of 3.2 mm and 3.7 mm diameters to simulate either a 'well-fitting' or a 'loosely fitting' situation. The RF values of the test implants were detected by the newly developed device which was directly mounted on the healing abutments of the implants. The results showed that the RF values of the implants under well-fitting conditions significantly increased (p < 0.01) 3 weeks after surgery and reached a plateau at around 6-7 weeks. Meanwhile implants with higher initial RF values had shorter healing times and higher final RF values at the plateau. Based on these findings, it was concluded that the idea of using the current designed device for detecting the degree of bone healing during the osseointegration process seems feasible.

The Callander laryngoscope blade modification is associated with a decreased risk of dental contact.

PURPOSE: Dental damage may occur with laryngoscopy. The purpose of this study was twofold: to determine whether preoperative examination could predict the risk of contacting the teeth with the laryngoscope and to evaluate the effectiveness of a modified Macintosh blade on reducing dental contact. METHODS: Four hundred and eighty-three patients scheduled for elective surgery requiring general anesthesia with endotracheal tube placement were studied prospectively. Features that might predict difficult intubation were assessed preoperatively. Laryngoscopy was performed twice on each patient, once with a regular Macintosh 3 blade and once with a blade in which the flange was partially removed (Callander modification). The distance between the flange of the blade and the upper incisors at glottic exposure was measured. We calculated correlations between individual airway characteristics and the chance of hitting the upper teeth with the regular Macintosh 3 blade and compared the frequencies of contacting the teeth between the two blades. RESULTS: The chance of hitting the upper teeth when using the regular Macintosh 3 blade increased significantly with non-parametric scores for Mallampati classification, mandibular subluxation, head and neck movement, interincisor gap, and condition of the upper teeth. (P < 0.01) The frequency of direct contact varied significantly between the two blades: 20.3% vs 4.1% for Macintosh 3 and modified blades, respectively (P < 0.05). Laryngeal views were improved with the modified blade. CONCLUSION: Airway characteristics correlate with the risk of hitting the upper teeth during laryngoscopy. The modified Macintosh blade reduces the risk of contacting the teeth.

Comparative allergenicity studies of native and recombinant Blomia tropicalis Paramyosin (Blo t 11).

BACKGROUND: The complementary DNA (cDNA) encoding for Blo t 11, a 102 kD allergen from Blomia tropicalis (Bt) was isolated, expressed and characterized previously. This study aimed to isolate the native Blo t 11 allergen and compare its allergenicity with the recombinant forms. METHODS: Native Blo t 11 (nBlo t 11) was isolated from crude Bt extract by immuno-affinity chromatography, analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, and verified by MALDI-TOF MS. Recombinant full-length Blo t 11 (rFL-Blo t 11) and its immunodominant peptide (fD) were expressed as glutathione S-transferase (GST)-fusion proteins in Escherichia coli. Immunoglobulin E (IgE) reactivity of the Blo t 11 allergens were determined by enzyme-linked immunosorbent assay (ELISA) and skin prick test. The inhibition capacity of the nBlo t 11 against fD and vice versa was determined by absorption studies. RESULTS: Affinity purified nBlo t 11 was susceptible to degradation with the major degraded product resolved at approximately 66 kD. The nBlo t 11 was confirmed by immunoblot analysis and MALDI-TOF MS that generated 13 peptides with complete identity to the deduced amino acid sequence of Blo t 11. Comparative in vitro and in vivo allergenicity tests and the cross inhibition studies between the native and recombinant Blo t 11 showed that recombinant fD, but not the rFL-Blo t 11, has comparable IgE reactivity with the native counterpart. CONCLUSIONS: This comparative study confirmed that the recombinant peptide fD contains the main immunodominant region of Blo t 11. This recombinant peptide, instead of the full-length protein, is a good candidate for diagnostic and therapeutics development for mite allergy.

Proteomic investigation of metabolic shift in mammalian cell culture.

Mammalian cells, under typical cultivation conditions, produce large quantities of lactate and ammonia that affect cell growth adversely and result in low cell concentration. Controlled nutrient feeding to maintain low concentrations of glucose and glutamine reduces metabolite production drastically, altering the metabolism of the cells. This metabolic shift results in higher cell concentration in continuous cultures and does not affect the specific productivity of the cells. We have taken a proteomics approach to investigate the differential protein expression with metabolic shift. Using two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS), we have found at least eight differentially expressed spots; two proteins were down-regulated, and the others were up-regulated with metabolic shift. These included metabolic enzymes, the brain form of phosphoglycerate mutase, which was down-regulated, and the precursor of the 23 kDa subunit of NADH-ubiquinone oxidoreductase, which was up-regulated. Another enzyme, the L1 isozyme of ubiquitin carboxyl-terminal hydrolase, which is involved in protein turnover and degradation, was also up-regulated in the metabolically altered cells. The remaining down-regulated spot had been identified as two isoforms of cytoplasmic actins, while three of the up-regulated spots were viral GAG polyproteins from various murine viruses. An unidentified protein was also up-regulated in the cells with altered metabolic state. This study shows the potential of using a proteomics approach in deciphering the intracellular changes in cells with physiological changes such as metabolism shift. The new insight into cell metabolism afforded by this analysis will greatly facilitate process optimization of continuous cell cultures.

Identification of a serine protease inhibitor homologue in Bird's Nest by an integrated proteomics approach.

For centuries, the edible nests of Collocalia spp. ("Bird's Nests") have been used as a Chinese delicacy that had been claimed to be an effective health-giving tonic. However, clinical studies indicated that in Singapore, Bird's Nest is the most common cause of food-induced anaphylaxis in children, which could lead to potentially life-threatening allergenic reactions. The purpose of this study was to characterize the major allergens in Bird's Nest by using the combined technologies of two-dimensional gel electrophoresis (2-DE), immunochemistry, N-terminal protein sequencing, and mass spectrometry. Results from the immunostaining of the Western blots of the Bird's Nest 2-DE separated proteins with the sera from allergic patients indicated the presence of a major allergen of 66 kDa. Initial searches of the matrix assisted laser desorption/ionization--time of flight--mass spectrometry (MALDI-TOF-MS) tryptic peptide masses of the allergen in the SWISS-PROT and NCBI nonredundant databases revealed that this protein was novel. Based on the partial protein sequence information obtained from N-terminal microsequencing and nanoelectrospray-tandem MS, the 66 kDa immunoreactive allergen was found to be homologous to ovoinhibitor, a Kazal-type serine protease inhibitor, which is one of the dominant allergens found in chicken egg white.

Proteome analysis of a human heptocellular carcinoma cell line, HCC-M: an update.

Recently, we reported the proteome analysis of a human hepatocellular carcinoma cell line, HCC-M (Electrophoresis 2000, 21, 1787-1813), using two-dimensional gel electrophoresis (2-DE) and matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). From a total of 408 unique spots excised from the 2-DE gel, 301 spots yielded good MALDI spectra. Out of these, 272 spots had matches returned from the database search leading to the identification of these proteins. Here, we report the results on the identification of the remaining 29 spots using nanoelectrospray ionization-tandem mass spectrometry (nESI-MS/MS). First, "peptide tag sequencing" was performed to obtain partial amino acid sequences of the peptides to search the SWISS-PROTand NCBI nonredundant protein databases. Spots that were still not able to find any matches from the databases were subjected to de novo peptide sequencing. The tryptic peptide sequences were used to search for homologues in the protein and nucleotide databases with the NCBI Basic Local Alignment Search Tool (BLAST), which was essential for the characterization of novel or post-translationally modified proteins. Using this approach, all the 29 spots were unambiguously identified. Among them, phosphotyrosyl phosphatase activator (PTPA), RNA-binding protein regulatory subunit, replication protein A 32 kDa subunit (RP-A) and N-acetylneuraminic acid phosphate synthase were reported to be cancer-related proteins.

Immunochemical characterization of edible bird's nest allergens.

BACKGROUND: We have previously described anaphylaxis induced by edible bird's nest (BN) and demonstrated that this condition is IgE mediated. OBJECTIVES: This study aimed at describing the immunochemical properties of the BN allergens. Comparative studies between 3 commercially available sources (according to the country of origin) of BN were also made. METHODS: Crude extracts of commercially available processed BN from Sarawak (Malaysia), Thailand, and Indonesia and fresh unprocessed BN from the caves of Sarawak were obtained by means of aqueous extraction. Specific IgE toward these sources were determined by using fluorescence allergosorbent tests (FASTs). Cross-reactivity studies between the 3 sources of commercially available processed BN were carried out by means of FAST inhibition. Immunochemical characterization by means of IgE immunoblot, periodate treatment, and heat stability studies were carried out on fresh unprocessed BN from Sarawak. RESULTS: Serum from allergic patients showed differences in IgE binding to the 3 sources of commercially available BN, with the highest levels of specific IgE recorded with the Sarawak source (P <.0001). Of these, only the Sarawak and Thailand sources showed considerable cross-reactivity. Further work on the unprocessed fresh Sarawak source identified a putative 66-kd major allergen containing several isoforms. Periodate treatment resulted in loss of IgE binding. Despite a progressive decline in the molecular weights of allergens on SDS-PAGE with increasing periods of boiling, IgE binding, as assessed by means of FAST, was not affected. N-terminal sequence of the major putative allergen (66 kd) showed homology to a domain of an ovoinhibitor precursor in chicken (SWISS-PROT accession No. P10184). CONCLUSIONS: We have described the immunochemical properties of BN allergens. Edible BN from different sources are allergenically dissimilar. The putative major allergen is a 66-kd protein.

An integrated approach in the discovery and characterization of a novel nuclear protein over-expressed in liver and pancreatic tumors.

An integrated approach in protein discovery through the use of multidisciplinary tools was reported. A novel protein, Hcc-1, was identified by analysis of the hepatocellular carcinoma (HCC)-M cell proteome. The assembled EST sequence of the 210 amino acid novel protein was subsequently confirmed by rapid amplification of cDNA ends (RACE). A total of 687 bp at the 5' untranslated region of Hcc-1 was identified. Promoter activity and several upstream open reading frames (uORFs) were demonstrated at this region. Bioinformatics prediction showed that the first 42 amino acids of the protein is a SAP domain with sequence matches to hnRNP from various vertebrate species. The Hcc-1 protein was localized to the cell nucleus while the gene was localized to chromosome 7q22.1. Hcc-1 cDNA level was increased in pancreatic adenocarcinoma. The level was also increased in well-differentiated hepatocellular carcinoma but decreases as the carcinoma progressed to a poorly differentiated stage.

Two-dimensional electrophoresis map of the human hepatocellular carcinoma cell line, HCC-M, and identification of the separated proteins by mass spectrometry.

Currently, one of the most popular applications of proteomics is in the area of cancer research. In Africa, Southeast Asia, and China, hepatocellular carcinoma is one of the most common cancers, occurring as one of the top five cancers in frequency. This project was initiated with the purpose of separating and identifying the proteins of a human hepatocellular carcinoma cell line, HCC-M. After two-dimensional gel electrophoresis separation, silver staining, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analyses, tryptic peptide masses were searched for matches in the SWISS-PROT and NCBI nonredundant databases. Approximately 400 spots were analyzed using this approach. Among the proteins identified were housekeeping proteins such as alcohol dehydrogenase, alpha-enolase, asparagine synthetase, isocitrate dehydrogenase, and glucose-6-phosphate 1-dehydrogenase. In addition, we also identified proteins with expression patterns that have been postulated to be related to the process of carcinogenesis. These include 14-3-3 protein, annexin, prohibitin, and thioredoxin peroxidase. This study of the HCC-M proteome, coupled with similar proteome analyses of normal liver tissues, tumors, and other hepatocellular carcinoma cell lines, represents the first step towards the establishment of protein databases, which are valuable resources in studies on the differential protein expressions of human hepatocellular carcinoma.

[Distribution of peptidergic neurons in the rat suprachiasmatic nucleus].

The distribution of the peptidergic neurons in the suprachiasmatic nucleus(SCN) in five adult male SD rats was examined by multi-immunolabelling for vasoactive intestinal polypeptide(VIP), arginine vasopressin (AVP) and somatostatin(SOM). The immunoreactions were visualized with an Elite ABC Kit(Vector Lab). The substrates in the peroxidase reaction were diaminobenzidine(DAB), tetramethylbenzidine(TMB) and 4-chloro-1-naphtol. The results showed that the differences in distribution and population of VIP-, AVP- and SOM-like neurons existed in the rostral, intermediate and caudal regions of the SCN and these neurons were overlapped and SOM-like neurons were scattered among them in the area between VIP- and AVP-like neurons. These findings may provide new morphologic evidences for explaining the functional mechanism in the SCN.

An improved synthesis of an umbelliferyl 5-thioxylopyranoside, precursor of the antithrombotic drug Iliparcil.

4-Ethyl-2-oxo-2H-1-benzopyran-7-yl 2,3,4-tri-O-acetyl-5-thio-beta-D-xylopyranoside, a synthetic intermediate of the orally active antithrombotic compound Iliparcil, has been prepared in 44-47% isolated yield. Different conditions were used for the glycosylation of 4-ethyl-2H-7-hydroxy-1-benzopyran-2-one 6 applying 2,3,4-tri-O-acetyl-5-thio-D-xylopyranosyl bromide (2), the analogous beta-chloride 3 or the alpha-trichloroacetimidate 5 as donors. With halides 2 and 3, the reaction was carried out in the presence of ZnO-ZnCl2 or ZnO alone. Both promoters are cheap, safe and therefore compatible with large-scale industrial processes.

High-throughput mass spectrometric discovery of protein post-translational modifications.

The availability of genome sequences, affordable mass spectrometers and high-resolution two-dimensional gels has made possible the identification of hundreds of proteins from many organisms by peptide mass fingerprinting. However, little attention has been paid to how information generated by these means can be utilised for detailed protein characterisation. Here we present an approach for the systematic characterisation of proteins using mass spectrometry and a software tool FindMod. This tool, available on the internet at http://www.expasy.ch/sprot/findmod.html , examines peptide mass fingerprinting data for mass differences between empirical and theoretical peptides. Where mass differences correspond to a post-translational modification, intelligent rules are applied to predict the amino acids in the peptide, if any, that might carry the modification. FindMod rules were constructed by examining 5153 incidences of post-translational modifications documented in the SWISS-PROT database, and for the 22 post-translational modifications currently considered (acetylation, amidation, biotinylation, C-mannosylation, deamidation, flavinylation, farnesylation, formylation, geranyl-geranylation, gamma-carboxyglutamic acids, hydroxylation, lipoylation, methylation, myristoylation, N -acyl diglyceride (tripalmitate), O-GlcNAc, palmitoylation, phosphorylation, pyridoxal phosphate, phospho-pantetheine, pyrrolidone carboxylic acid, sulphation) a total of 29 different rules were made. These consider which amino acids can carry a modification, whether the modification occurs on N-terminal, C-terminal or internal amino acids, and the type of organisms on which the modification can be found. We illustrate the utility of the approach with proteins from 2-D gels of Escherichia coli and sheep wool, where post-translational modifications predicted by FindMod were confirmed by MALDI post-source decay peptide fragmentation. As the approach is amenable to automation, it presents a potentially large-scale means of protein characterisation in proteome projects.

Towards an automated approach for protein identification in proteome projects.

The development of automated, high throughput technologies for the rapid identification of proteins is essential for large-scale proteome projects. While a degree of automation already exists in some stages of the protein identification process, such as automated acquisition of matrix assisted laser desorption ionisation-time of flight (MALDI-TOF) mass spectra, efficient interfaces between different stages are still lacking. We report the development of a highly automated, integrated system for large scale identification of proteins separated by two-dimensional gel electrophoresis (2-DE), based on peptide mass fingerprinting. A prototype robotic system was used to image and excise 288 protein spots from an amido black stained polyvinylidene difluoride (PVDF) blot. Protein samples were enzymatically digested with a commercial automated liquid handling system. MALDI-TOF mass spectrometry was used to acquire mass spectra automatically, and the data analysed with novel automated peptide mass fingerprinting database interrogation software. Using this highly automated system, we were able to identify 95 proteins on the basis of peptide mass fingerprinting, isoelectric point and molecular weight, in a period of less than ten working days. Advantages, problems, and future developments in robotic excision systems, liquid handling, and automated database interrogation software are discussed.