Evaluation of digital construction, production and intraoral position accuracy of novel 3D CAD/CAM titanium retainers.

OBJECTIVES: New opportunities have arisen to manufacture three-dimensional computer-aided design/computer-aided manufacturing (3D CAD/CAM) retainers from titanium blocks by digital cutting technology. These novel technologies need to fulfill requirements regarding digital planning and position accuracy. The aim of the present study was to investigate the digital construction, the CAD/CAM production and the intraoral positioning accuracy of custom-manufactured novel 3D CAD/CAM titanium retainers. MATERIALS AND METHODS: A total of 37 prime4me® RETAIN3R (Dentaurum, Ispringen, Germany) retainers were inserted to stabilize the upper anterior front teeth. Following insertion, an intraoral scan was used to record the position. The intraoral position was compared to the virtual setup using 3D superimposition software. Measurement points were evaluated in all three dimensions (horizontal, sagittal and vertical planes). Data were analyzed using Kruskal-Wallis test followed by Dunn's multiple comparison test. RESULTS: A total of 185 measurements were performed. The horizontal plane and the sagittal plane demonstrated a high level of positioning accuracy between the planned and the intraoral position. Statistically significant deviations between the preceding virtual setup and the intraoral situation were observed in the vertical dimension. Within the retainer, the intraoral positioning accuracy decreased for the measurement points in the direction of the distal retainer segment. CONCLUSION: Based on the results, the present study shows a high level of congruence between the 3D virtually planning and the final intraoral position of the fabricated novel 3D CAD/CAM titanium retainers.

Electrodialytic removal of tungsten and arsenic from secondary mine resources - Deep eutectic solvents enhancement.

Tungsten is a critical raw material for European and U.S. economies. Tungsten mine residues, usually considered an environmental burden due to e.g. arsenic content, are also secondary tungsten resources. The electrodialytic (ED) process and deep eutectic solvents (DES) have been successfully and independently applied for the extraction of metals from different complex environmental matrices. In this study a proof of concept demonstrates that coupling DES in a two-compartment ED set-up enhances the removal and separation of arsenic and tungsten from Panasqueira mine secondary resources. Choline chloride with malonic acid (1:2), and choline chloride with oxalic acid (1:1) were the DES that in batch extracted the average maximum contents of arsenic (16%) and tungsten (9%) from the residues. However, when ED was operated at a current intensity of 100 mA for 4 days, the extraction yields increased 22% for arsenic and 11% for tungsten, comparing to the tests with no current. From the total arsenic and tungsten extracted, 82% and 77% respectively were successfully removed from the matrix compartment, as they electromigrated to the anolyte compartment, from where these elements can be further separated. This achievement potentiates circular economy, as the final treated residue could be incorporated in construction materials production, mitigating current environmental problems in both mining and construction sectors.

High expression of human carboxypeptidase M in Pichia pastoris: purification and partial characterization.

Carboxypeptidase M (CPM) is an extracellular glycosylphosphatidyl-inositol-anchored membrane glycoprotein, which removes the C-terminal basic residues, lysine and arginine, from peptides and proteins at neutral pH. CPM plays an important role in the control of peptide hormones and growth factor activity on the cell surface. The present study was carried out to clone and express human CPM in the yeast Pichia pastoris in order to evaluate the importance of this enzyme in physiological and pathological processes. The cDNA for the enzyme was amplified from total placental RNA by RT-PCR and cloned in the vector pPIC9, which uses the methanol oxidase promoter and drives the expression of high levels of heterologous proteins in P. pastoris. The cpm gene, after cloning and transfection, was integrated into the yeast genome, which produced the active protein. The recombinant protein was secreted into the medium and the enzymatic activity was measured using the fluorescent substrate dansyl-Ala-Arg. The enzyme was purified by a two-step protocol including gel filtration and ion-exchange chromatography, resulting in a 1753-fold purified active protein (16474 RFU mg protein(-1) min(-1)). This purification protocol permitted us to obtain 410 mg of the purified protein per liter of fermentation medium. SDS-PAGE showed that recombinant CPM migrated as a single band with a molecular mass similar to that of native placental enzyme (62 kDa), suggesting that the expression of a glycosylated protein had occurred. These results demonstrate for the first time the establishment of a method using P. pastoris to express human CPM necessary to the development of specific antibodies and antagonists, and the analysis of the involvement of this peptidase in different physiological and pathological processes.

The role of nt590 P21 gene polymorphism in the susceptibility to nasopharyngeal cancer.

AIM: The purpose of this study was to assess if the P21 nt590 polymorphism is associated with the susceptibility to nasopharyngeal cancer and with the age at diagnosis. MATERIALS AND METHODS: We analyzed the frequency of 3'UTR P21 polymorphisms in blood samples from 102 nasopharyngeal cancer patients and 191 controls, with no known oncologic disease, using PCR-RFLP. RESULTS: The polymorphism genotype frequencies were 93.2% (CC), 5.2% (CT) and 1.6% (TT) in the control group and 88.2% (CC), 10.8% (CT) and 1.0% (TT) in the cases group. We found no statistically significant association between the different P21 polymorphism genotypes and risk of nasopharyngeal cancer (p = 0.201). However, approximately a four-fold increased risk of undifferentiated nasopharyngeal carcinoma in early stages was observed for P21 T carriers (OR = 3.734; 95% IC 1.289-10.281; p = 0.01). Furthermore, our results indicate that the waiting time for onset of neoplasia in T carriers patients was 12.4 years earlier (56.5 years old), comparing with those carrying CC genotype (68.9 years old). CONCLUSIONS: Our findings suggest that the 3'UTR P21 polymorphism may play an important role in the pathogenesis and initiation, but not in the progression, of undifferentiated nasopharyngeal carcinoma. Moreover, the polymorphism seems to contribute to a significantly earlier age at diagnosis.

High expression of human carboxypeptidase M in Pichia pastoris: Purification and partial characterization

Carboxypeptidase M (CPM) is an extracellular glycosylphosphatidyl-inositol-anchored membrane glycoprotein, which removes the C-terminal basic residues, lysine and arginine, from peptides and proteins at neutral pH. CPM plays an important role in the control of peptide hormones and growth factor activity on the cell surface. The present study was carried out to clone and express human CPM in the yeast Pichia pastoris in order to evaluate the importance of this enzyme in physiological and pathological processes. The cDNA for the enzyme was amplified from total placental RNA by RT-PCR and cloned in the vector pPIC9, which uses the methanol oxidase promoter and drives the expression of high levels of heterologous proteins in P. pastoris. The cpm gene, after cloning and transfection, was integrated into the yeast genome, which produced the active protein. The recombinant protein was secreted into the medium and the enzymatic activity was measured using the fluorescent substrate dansyl-Ala-Arg. The enzyme was purified by a two-step protocol including gel filtration and ion-exchange chromatography, resulting in a 1753-fold purified active protein (16474 RFU mg protein-1 min-1). This purification protocol permitted us to obtain 410 mg of the purified protein per liter of fermentation medium. SDS-PAGE showed that recombinant CPM migrated as a single band with a molecular mass similar to that of native placental enzyme (62 kDa), suggesting that the expression of a glycosylated protein had occurred. These results demonstrate for the first time the establishment of a method using P. pastoris to express human CPM necessary to the development of specific antibodies and antagonists, and the analysis of the involvement of this peptidase in different physiological and pathological processes.