[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.

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.

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.

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.

Two distinct forms of the peridinin-chlorophyll a-protein from Amphidinium carterae.

Peridinin-chlorophyll a-proteins (PCPs) have been purified by combination of ammonium sulphate precipitation and cation exchange chromatography. The amino acid sequences of several of the most abundant forms have been deduced by direct protein sequencing and from DNA and indicate a highly conserved multi-gene family. At least two of the PCP genes are tandemly arranged. A novel form of the protein was also obtained in low yield with fewer peridinins (six vs eight) per chlorophyll a and with a different molecular mass (34 kDa vs 32 kDa) of its apoprotein. It had only 31% sequence identity with any of the more abundant PCP forms but retained a two-domain structure.

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.

Protein identification with N and C-terminal sequence tags in proteome projects.

Genome sequences are available for increasing numbers of organisms. The proteomes (protein complement expressed by the genome) of many such organisms are being studied with two-dimensional (2D) gel electrophoresis. Here we have investigated the application of short N-terminal and C-terminal sequence tags to the identification of proteins separated on 2D gels. The theoretical N and C termini of 15, 519 proteins, representing all SWISS-PROT entries for the organisms Mycoplasma genitalium, Bacillus subtilis, Escherichia coli, Saccharomyces cerevisiae and human, were analysed. Sequence tags were found to be surprisingly specific, with N-terminal tags of four amino acid residues found to be unique for between 43% and 83% of proteins, and C-terminal tags of four amino acid residues unique for between 74% and 97% of proteins, depending on the species studied. Sequence tags of five amino acid residues were found to be even more specific. To utilise this specificity of sequence tags for protein identification, we created a world-wide web-accessible protein identification program, TagIdent (http://www.expasy.ch/www/tools.html), which matches sequence tags of up to six amino acid residues as well as estimated protein pI and mass against proteins in the SWISS-PROT database. We demonstrate the utility of this identification approach with sequence tags generated from 91 different E. coli proteins purified by 2D gel electrophoresis. Fifty-one proteins were unambiguously identified by virtue of their sequence tags and estimated pI and mass, and a further 11 proteins identified when sequence tags were combined with protein amino acid composition data. We conlcude that the TagIdent identification approach is best suited to the identification of proteins from prokaryotes whose complete genome sequences are available. The approach is less well suited to proteins from eukaryotes, as many eukaryotic proteins are not amenable to sequencing via Edman degradation, and tag protein identification cannot be unambiguous unless an organism's complete sequence is available.

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.

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.

(Benzoylphenyl)piperidines: a new class of immunomodulators.

A series of (benzoylphenyl)piperidines has been synthesized and evaluated for activity as immunomodulators. Several of these compounds show good activity in primary screening on the basis of the lymphocytes mitogenic response to Con A, PHA, and PWM. A chloro group in position 4 of the benzoyl moiety as well as an amino group (or a carbamate derivative) para to the piperidine nucleus seems to be essential for activity. The depicted compounds may be considered as the first examples of a new series of immunomodulators.

Induction of Fc and C3b receptors on rabbit corneal cells by herpes simplex virus.

The expression of receptors for Fc portion (FcR) of immunoglobulin G (IgG) and for a C3b component of complement (C3bR) by herpes simplex virus (HSV) was studied in primary cultures of rabbit corneal cells. Monolayer cultures of epithelial, stromal and endothelial cells of the rabbit cornea were infected with three strains each of type 1 (HSV-1) and type 2 HSV (HSV-2). Rosette methods were used to detect receptors by means of sheep erythrocytes sensitized with rabbit IgG for FcR and C3b-coated sheep erythrocytes for C3bR. The FcR were expressed regularly on epithelial, stromal and endothelial cells by all three strains of both HSV-1 and HSV-2. The C3bR, however, were expressed only by HSV-1 on epithelial and stromal cells. Little or no C3bR activities could be detected on endothelial cells infected with any strain of HSV-1 or HSV-2. The FcR and C3bR expressed on corneal cells were induced by HSV and were blocked by monoclonal antibody to HSV-1 glycoprotein E(gE) or glycoprotein C(gC) respectively, confirming findings of other investigators that gE acts as FcR and gC as C3bR.

Role of virus-infected mononuclear leukocytes in herpetic chorioretinitis of newborn rabbits.

The role of virus-infected mononuclear leukocytes (MNLs) in the pathogenesis of neonatal herpetic chorioretinitis in newborn rabbits was investigated. As early as 2 days after inoculating the animals' skins with type 2 herpes simplex virus (HSV-2), infectious MNLs in the infected animals' peripheral blood were found. The virus was associated, for the most part, with MNLs that belonged to phagocytic and adherent cell fractions. Observations by electron microscopy indicated that HSV-2 was actively replicating in the MNLs. It was also found that as few as 80 virus-infected MNLs injected via the right common carotoid artery were capable of inducing the chorioretinal lesions in 50% of the eyes, but that as many as 10(3) Pfu of free virus were required to produce the same lesions in the same percentage of eyes. This result clearly indicated that virus-infected MNLs were far more efficient in producing chorioretinitis than free virus, and may thus play a crucial role in the pathogenesis of herpetic chorioretinitis in newborn rabbits. When 111In-labeled virus-infected or uninfected MNLs were injected into normal rabbits via the right common carotid artery, the virus-infected MNLs localized more readily in the eye than the uninfected MNLs. The virus-infected MNLs also attached to the cultured vascular endothelial cells significantly more often than the uninfected MNLs. These results suggested that virus-infected MNLs might be easily trapped in the circulation of the eye and, in this way, produce the ocular lesions.

Large-scale amino-acid analysis for proteome studies.

Amino-acid analysis is a relatively new method for identification of proteins separated by two-dimensional gel electrophoresis and blotted onto polyvinylidene difluoride (PVDF) membranes. This article describes modified amino-acid analysis methods for this purpose. Streamlined sample handling is a key feature of the process. To minimise sample manipulation, a single vial is used for hydrolysis and the protein hydrolysate on PVDF membrane is extracted by a one-step procedure. The hydrolysate should not be stored for long periods before analysis. Applications of the technique are presented to demonstrate the identification procedure. This approach is the most cost-effective and time-effective first step in mass protein screening for a large-scale proteome project.

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.

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.

From proteins to proteomes: large scale protein identification by two-dimensional electrophoresis and amino acid analysis.

Separation and identification of proteins by two-dimensional (2-D) electrophoresis can be used for protein-based gene expression analysis. In this report single protein spots, from polyvinylidene difluoride blots of micropreparative E. coli 2-D gels, were rapidly and economically identified by matching their amino acid composition, estimated pI and molecular weight against all E. coli entries in the SWISS-PROT database. Thirty proteins from an E. coli 2-D map were analyzed and identities assigned. Three of the proteins were unknown. By protein sequencing analysis, 20 of the 27 proteins were correctly identified. Importantly, correct identifications showed unambiguous "correct" score patterns. While incorrect protein identifications also showed distinctive score patterns, indicating that protein must be identified by other means. These techniques allow large-scale screening of the protein complement of simple organisms, or tissues in normal and disease states. The computer program described here is accessible via the World Wide Web at URL address (http:@expasy.hcuge.ch/).

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.