Prosthetic-Driven Autotransplantation with the Assistance of Medical Image-Processing Software and a Real-Time Navigation System: A Case Report.

Autotransplantation has been proven as a viable method of reconstructing missing teeth. While preparing the recipient site, the bone reduction location depends largely on the surgeon's experience. Inappropriate overpreparation can cause biologic and esthetic complications, such as buccal or lingual bone resorption. This paper provides an innovative method to aid clinicians in precisely preparing a recipient site with the assistance of medical image-processing software and a real-time navigation system. This case report presents the autotransplantation of a mandibular molar using this technique with good short-term (6 months) clinical outcomes, including radiographic bone fill, normal probing pocket depth, physiologic tooth mobility, acceptable gingival level, and satisfactory restoration.

A Socket Seal Technique With the Use of Autologous Dental Roots for Socket Seal: A Case Series.

The literature identified variations in socket seal surgery, each with limitations. This case series aimed to observe the outcome of using autologous dental root (ADR) for socket sealing on socket preservation (SP). A total of 9 patients with 15 extraction sockets were documented. After flapless extraction, the xenograft or alloplastic grafts were placed in the sockets. Autologous dental roots were prepared extraorally and applied to seal the socket entrance. All SP sites healed uneventfully. Cone-beam computed tomography (CBCT) scan was performed after 4-6 months of healing to evaluate ridge dimensions. The preserved alveolar ridge profiles were verified on CBCT scans and during implant surgery. Implants were placed successfully with a reduced need for guided bone regeneration. Histological biopsy specimens were examined in 3 cases. The histological examination demonstrated vital bone formation and osseointegration of graft particles. All patients completed the final restorations and were monitored for 15.56 ± 9.08 months after functional loading. The favorable clinical outcomes support the use of ADR for SP procedures. It was not only accepted to patients but also easy to perform with low complication rates. The ADR technique is thus a feasible method for socket seal surgery.

Convergent angles of a tapered implant referred from the root profile of premolars.

Background/purpose: Limited studies have discussed the convergent profiles regarding tapered implants based on biological considerations. This study analyzed the convergent angles (CAs) of premolar roots and imitated a tapered implant according to the anatomy of tooth roots. Materials and methods: A total of 60 single-rooted premolars were explored by micro-computed tomography. Every individual root was divided into 10 segments corono-apically, and the roots' buccolingual (BL) and mesiodistal (MD) CAs were measured by sections. To mimic a dental implant, the irregular shape of examined root cross-sections was transformed into a circular shape with equal areas. A biomimetic dental implant (BDI) was reconstructed and its CAs were compared with those of the natural roots' BL and MD at the examined levels and overall estimation. Results: In general, the maxillary and mandibular premolars demonstrated comparable CA patterns. However, significantly different CA patterns of BL, MD, and BDI were developed for both the maxillary and mandibular roots at the examined levels. The BL's CAs were greater than those CAs measured from the BDI and MD aspects, particularly for the sections at the middle and apical thirds of the roots. For overall CAs, the BDI's CAs were comparable with the average CAs of the BL and MD for both premolar groups. Conclusion: Instead of a cylindrical configuration, the BDI prototype demonstrated a tapered model with a continuous slope. The average CA of BDI was 14°-24°, serving as a biological reference for future tapered implant design and research.

Physiological reaction of anxious patients taking sedative medications before and after periodontal surgery.

Background/purpose: Clinicians use sedatives for anxiety patients at times in daily practice, but the direct influence of the medication on the wound healing of periodontal tissues is unknown. The aim of this study was to analyze the influence of the short-term administration of diazepam to patients with dental anxiety undergoing free gingival graft (FGG) procedures. Materials and methods: A total of 51 FGG procedures in 39 patients were included. Twenty-six anxious patients medicated with 5 mg of diazepam from the night before surgery to 7 days after surgery served as the medication group, and the rest served as the control group. Direct examination, photographs and H2O2 were used to evaluate the healing of palatal wounds. Stress levels and sleep quality, and salivary melatonin levels were assessed. Results: On Day 14, complete epithelization of the wounds was noted in 48.39% of the patients in the medication group and 35.29% of the patients in the control group. Regardless of whether they receive medication or not, groups with complete epithelialization by Day 14 had higher levels of preoperative melatonin than those without, with a P value of 0.02. The postoperative melatonin in the medication group tended to present higher levels than the control group. Conclusion: Higher preoperative melatonin levels can accelerate wound healing. The short-term administration of the diazepam seemed to facilitate palatal wound healing by reducing stress and maintaining postoperative melatonin levels. This is the first time the relationships between sedatives, melatonin levels and palatal wound healing has been reported.

Efficacy of free gingival grafting to augment keratinized mucosa around dental implants in posterior regions after restorative procedures: A retrospective clinical study.

STATEMENT OF PROBLEM: Data on the shrinkage of free gingival grafts (FGGs) vary. Most studies have analyzed grafts in nonmolar sites because of measurement limitations and have addressed the changes in grafts and keratinized mucosa width (KMW) only in the early healing phase. PURPOSE: The purpose of this retrospective clinical study was to assess the dimensional changes of an FGG in the posterior regions and their influencing factors, with the aim of obtaining sufficient and stable KMW after restoration. MATERIAL AND METHODS: A total of 77 implants in 40 participants who had undergone an FGG surgery were recruited. Graft sizes during surgery and the surface areas of keratinized mucosa at the follow-up visit after restorations were compared by digital analysis and verified by clinical measurements and photographs. The association between shrinkage and the graft sizes, implant location, and sex and age of the participants was evaluated. The influence of the shrinkage of FGG on the KMW after restoration was analyzed by multivariable linear regression with generalized estimating equation (GEE) models. RESULTS: The mean ±standard deviation shrinkage of FGG around implants in the posterior regions was 24.76 ±14.77%, and the mean ±standard deviation KMW was 4.16 ±1.77 mm at the follow-up visit. Larger grafts had a statistically higher shrinkage ratio (P<.001). No statistically significant difference was found regarding the effect of implant location, sex, and age on the shrinkage of FGG and final KMW (P>.05). The mean ±standard deviation follow-up period after restoration was 12.45 ±7.73 months CONCLUSIONS: Free gingival grafting was found to be a predictable treatment approach for augmentation of KMW around implants in the posterior region after the fabrication of prostheses as long as grafts of sufficient size were placed. Stable outcomes were shown in the study participants in the follow-up period of up to 3 years.

Morphology of Peri-Implant Tissues Around Permanent Prostheses With Various Emergence Angles Following Free Gingival Grafting.

PURPOSE: To analyze the tissue morphology around implant-supported prostheses by digital technology and to evaluate the effect of prosthetic contours on the changes in tissues following the free gingiva graft procedure. MATERIAL AND METHODS: A total of 53 implants in 32 patients receiving free gingiva grafts were selected. These had previously presented insufficient keratinized mucosa width (KMW). At the follow-up visits (mean: 16.66 ± 9.97 months), the implant position and tissue condition were documented with an oral scanner. Vertical soft tissue thickness (VT), measured from the implant-abutment connection to the marginal tissues, and horizontal soft tissue thickness (HT), at the level of the platform, were calculated. The VT, HT, and emergence angle (EA) of prostheses were assessed by 3Shape analyzing software. The final KMW was measured by clinical assessment. Marginal bone loss (MBL) was calculated in the follow-up bitewing radiographs. RESULTS: The mean VT in the study was 2.65 ± 0.75 mm at the mid-buccal sites, 3.74 ± 1.22 mm at the mesial, 3.16 ± 1.08 mm at the distal, and 2.53 ± 0.92 at the mid-lingual aspects. The mid-buccal HT was 1.45 ± 0.53 mm while the mid-lingual was 1.05 ± 0.43 mm (p = 0.008). Interestingly, prostheses with mid-buccal EA > $\; > \;$ 30° exhibited slightly lower VT, but higher HT, than the ones with EA ≤ $\; \le \;$ 30°. Prostheses with proximal EA > 30° displayed slightly more MBL, compared to prostheses with EA ≤ $\; \le \;$ 30°. The mean KMW was 4.08 ± 1.10 mm. CONCLUSIONS: Free gingival grafting is a predictable treatment approach to augmenting soft tissue 3-dimensionally. Prostheses with EA ≤ $\; \le \;$ 30° were preferable for preserving the maximal VT and maintaining crestal bone stability.

Decoronation-induced infected alveolar socket defect rat model for ridge preservation.

Current rat alveolar ridge preservation models have not been well standardized. In this study, we proposed decoronation-induced infected alveolar socket model of rat. The bilateral maxillary first molars (M1) of twenty-four rats were decoronized or extracted. After 2, 6, 10, and 14 weeks, bone and soft tissue changes at M1 and periodontal conditions of maxillary second (M2) and third molars (M3) were evaluated by micro-computed tomography and histological analysis. Additional eighteen rats with standardized size defects were grafted with Bio-Oss Collagen to compare with unmanipulated contralateral side. Decoronation preserved greater bone and soft tissue dimensions at M1, provided larger three-dimensional (3D) bone contour volume, but also promoted periodontal breakdown of M2 Histological results showed intense inflammatory cell infiltrations and severe bone resorption within M1 socket and at mesial aspect of M2. The critical dimensions to accommodate largest standardized defect at M1 were 2.2-2.3 mm at vertical bone height and 2.8-3.2 mm at alveolar crestal width. Bio-Oss Collagen could not fully preserve buccal or palatal bone height but could be beneficial in preserving ridge width in large alveolar defects. Collectively, if periodontally-involved alveolar bone defect is preferred, we suggest extracting M1 roots 6 weeks after decoronation to allow periodontitis to occur at M2. If standardized critical dimension defect is preferred, we suggest extracting M1 roots 2 weeks after decoronation, and creating defect in the middle of M1 site with size no larger than 2.7 mm diameter to its full depth.

Metformin-Incorporated Gelatin/Nano-Hydroxyapatite Scaffolds Promotes Bone Regeneration in Critical Size Rat Alveolar Bone Defect Model.

In this study, we fabricated gelatin/nano-hydroxyapatite/metformin scaffold (GHMS) and compared its effectiveness in bone regeneration with extraction-only, Sinbone, and Bio-Oss Collagen® groups in a critical size rat alveolar bone defect model. GHMS was synthesized by co-precipitating calcium hydroxide and orthophosphoric acid within gelatin solution, incorporating metformin, and cross-linked by microbial transglutaminase. The morphology, characterization, and biocompatibility of scaffold were examined. The in vitro effects of GHMS on osteogenic gene and protein expressions were evaluated. In vivo bone formation was assessed in a critical size rat alveolar bone defect model with micro-computed tomography and histological examination by comparing GHMS with extraction-only, Sinbone, and Bio-Oss Collagen®. The synthesized GHMS had a highly interconnected porous structure with a mean pore size of 81.85 ± 13.8 µm. GHMS exhibited good biocompatibility; promoted ALPL, RUNX2, SP7, BGLAP, SPARC and Col1a1 gene expressions; and upregulated the synthesis of osteogenic proteins, including osteonectin, osteocalcin, and collagen type I. In critical size rat alveolar bone defects, GHMS showed superior bone regeneration compared to extraction-only, Sinbone, and Bio-Oss Collagen® groups as manifested by greater alveolar ridge preservation, while more bone formation with a lower percentage of connective tissue and residual scaffold at the defect sites grafted with GHMS in histological staining. The GHMS presented in this study may be used as a potential bone substitute to regenerate alveolar bone. The good biocompatibility, relatively fast degradation, interconnected pores allowing vascularization, and higher bioactivity properties of the components of the GHMS (gelatin, nHA, and metformin) may contribute to direct osteogenesis.

Restoration of a wide edentulous posterior site with two small-diameter implants: Biologically-driven alternative treatment.

BACKGROUND/PURPOSE: Crestal bone stability, implant rigidity and occlusal loading are issues with small-diameter implants. This article demonstrates the use of two small-diameter implants replacing a missing wide edentulous site and discusses factors that may affect bone changes. METHODS: Patients who wanted to restore an edentulous space measuring from 12 to 14 mm wide in the posterior region were offered an alternative treatment option, using two narrow or regular-diameter implants instead of one wide implant. In the study, the crestal bone stability of 12 implants in 6 edentulous sites was assessed by cone beam CTs and periapical radiographs in follow-up visits for up to 4 years. RESULTS: The bone level of all the implants was stable at buccal, lingual, mesial and distal sites, with mean values < 1 mm. The average buccal bone thickness was 1.15 ± 1.07 mm and lingual was 1.86 ± 0.89 mm, meaning that implants were surrounded by a sufficient amount of bone. The good treatment outcome may be attributed to the capability of fabricating better emergence profiles, angles (Mean: 20.67 ± 7.82° at the mesial and 20.25 ± 8.23° at the distal site) and cleansable embrasures of prostheses which are key to maintaining good oral hygiene and implant health. CONCLUSION: Using two narrow or regular-diameter implants to replace a single edentulous site measured around 12-14 mm wide in posterior region seemed to be a feasible treatment option. It is especially suitable for sites with ridge atrophy and/or patients suffering from systemic diseases.

Molecular Cloning of Mouse Homologue of Enamel Protein C4orf26 and Its Phosphorylation by FAM20C.

It is widely accepted that cellular processes are controlled by protein phosphorylation and has become increasingly clear that protein degradation, localization and conformation as well as protein-protein interaction are the examples of subsequent cellular events modulated by protein phosphorylation. Enamel matrix proteins belong to members of the secretory calcium binding phosphoprotein (SCPP) family clustered on chromosome 4q21, and most of the SCPP phosphoproteins have at least one S-X-E motifs (S; serine, X; any amino acid, E; glutamic acid). It has been reported that mutations in C4orf26 gene, located on chromosome 4q21, are associated with autosomal recessive type of Amelogenesis Imperfecta (AI), a hereditary condition that affects enamel formation/mineralization. The enamel phenotype observed in patients with C4orf26 mutations is hypomineralized and partially hypoplastic, indicating that C4orf26 protein may function at both secretory and maturation stages of amelogenesis. The previous in vitro study showed that the synthetic phosphorylated peptide based on C4orf26 protein sequence accelerates hydroxyapatite nucleation. Here we show the molecular cloning of Gm1045, mouse homologue of C4orf26, which has 2 splicing isoforms. Immunohistochemical analysis demonstrated that the immunolocalization of Gm1045 is mainly observed in enamel matrix in vivo. Our report is the first to show that FAM20C, the Golgi casein kinase, phosphorylates C4orf26 and Gm1045 in cell cultures. The extracellular localization of C4orf26/Gm1045 was regulated by FAM20C kinase activity. Thus, our data point out the biological importance of enamel matrix-kinase control of SCPP phosphoproteins and may have a broad impact on the regulation of amelogenesis and AI.

Retrospective evaluation of gingival thickness and tissue stability after connective tissue grafting in Asian populations: 1 month to 3.5 years.

BACKGROUND/PURPOSE: The objectives of this retrospective study are to analyze post-surgical gingival thickness after connective tissue grafting in the Asian population and to assess its tissue stability for up to approximately 3.5 years. METHODS: A total of 111 grafted teeth and 57 nearby nongrafted teeth in 28 Asian patients who had undergone connective tissue grafting surgery were selected. Gingival thickness was measured by transgingival probing. The mean gingival thickness of the grafted teeth was compared with adjacent nongrafted teeth in the same individuals. The mean gingival thickness of the grafted teeth in different tooth types and at various time intervals were statistically analyzed. RESULTS: The average gingival thickness following connective tissue grafting is 1.99 ± 0.62 mm compared to 0.96 ± 0.40 mm with nongrafted teeth (P < .0001). The maxillary premolar is the tooth type that underwent connective tissue grafting most frequently in our study. Among different tooth types, mandibular molars showed the thickest gingival tissues whereas mandibular incisors presented the thinnest tissues. No statistically significant difference in the mean tissue thickness at different time intervals was observed. CONCLUSION: Connective tissue grafting is a predictable treatment modality for gingival phenotype conversion, even in Asians, achieving nearly 2 mm of gingival thickness on average, post-operation. Tissue stability after connective tissue grafting has been presented in our study. This quantitative assessment of the gingival thickness in Asians may encourage clinicians to deal with soft tissue architecture ahead of main surgical, restorative and orthodontic treatments in order to achieve pleasing treatment outcomes.

FAM20C directly binds to and phosphorylates Periostin.

It is widely accepted that FAM20C functions as a Golgi casein kinase and has large numbers of kinase substrates within the secretory pathway. It has been previously reported that FAM20C is required for maintenance of healthy periodontal tissues. However, there has been no report that any extracellular matrix molecules expressed in periodontal tissues are indeed substrates of FAM20C. In this study, we sought to identify the binding partner(s) of FAM20C. FAM20C wild-type (WT) and its kinase inactive form D478A proteins were generated. These proteins were electrophoresed and the Coomassie Brilliant Blue (CBB)-positive bands were analyzed to identify FAM20C-binding protein(s) by Mass Spectrometry (MS) analysis. Periostin was found by the analysis and the binding between FAM20C and Periostin was investigated in cell cultures and in vitro. We further determined the binding region(s) within Periostin responsible for FAM20C-binding. Immunolocalization of FAM20C and Periostin was examined using mouse periodontium tissues by immunohistochemical analysis. In vitro kinase assay was performed using Periostin and FAM20C proteins to see whether FAM20C phosphorylates Periostin in vitro. We identified Periostin as one of FAM20C-binding proteins by MS analysis. Periostin interacted with FAM20C in a kinase-activity independent manner and the binding was direct in vitro. We further identified the binding domain of FAM20C in Periostin, which was mapped within Fasciclin (Fas) I domain 1-4 of Periostin. Immunolocalization of FAM20C was observed in periodontal ligament (PDL) extracellular matrix where that of Periostin was also immunostained in murine periodontal tissues. FAM20C WT, but not D478A, phosphorylated Periostin in vitro. Consistent with the overlapped expression pattern of FAM20C and Periostin, our data demonstrate for the first time that Periostin is a direct FAM20C-binding partner and that FAM20C phosphorylates Periostin in vitro.

Interdisciplinary Approach to Treat Patients With Tooth Agenesis: A Case Report.

INTRODUCTION: Tooth agenesis is a type of dental anomaly in which individuals are missing teeth due to developmental failure as a result of genetic or environmental factors. With approximately one fourth of the population missing ≥1 third molar, tooth agenesis is considered a common dental anomaly. However, the severity of tooth agenesis can range from a missing single tooth to multiple teeth. When suffering from severe tooth agenesis, the patient's health and social relationships are often affected. CASE PRESENTATION: The patient in this report congenitally lost 11 teeth and suffered from compromised esthetics and impaired chewing function. In such a severe tooth agenesis case, interdisciplinary treatments involving orthodontics, periodontics, and prosthodontics were engaged to reconstruct the ideal biology, function, and esthetics for the patient. With an interdisciplinary approach, the periodontist played an important role in the rehabilitation of the edentulous regions with implants in combination with various hard and soft tissue augmentation procedures. In addition, the patient with severe tooth agenesis presented with additional dental anomalies. The periodontist, therefore, had to collaborate with other specialists to provide early detection and intervention to avoid future complications, such as the management of infraoccluded ankylosed deciduous molars and aberrant frenum. The patient at the end of treatment had a good occlusion with improved function and esthetics. CONCLUSION: This case report describes the interdisciplinary treatment approach used and points out the role of periodontists in the treatment of a patient with severe tooth agenesis.

Staged orthodontic treatment in preparation for immediate implant placement: A clinical report with a 5-year follow-up.

Anterior implant restoration is one of the most challenging restorative procedures, especially for sites with vertical and/or horizontal hard and soft tissue deformities. Orthodontic extrusion before implant placement may be the best means of overcoming vertical deficiencies. This article describes a modification to the standard technique, involving staged orthodontic extrusion and buccal root torque. The main advantage of this modification is that it encourages bone and soft tissue development, thereby allowing the patient to receive immediate or early implant treatment. A clinical procedure is presented to illustrate the modified technique that resulted in an esthetically pleasing and stable 5-year outcome.

10-hydroxy-2-decenoic acid of royal jelly exhibits bactericide and anti-inflammatory activity in human colon cancer cells.

BACKGROUND: Royal jelly (RJ), the exclusive food for the larva of queen honeybee, is regarded as the novel supplement to promote human health. The function of RJ may be attributed to its major and unique fatty acid, 10-hydroxy-2-decenoic acid (10-HDA). The current study investigated the anti-inflammory function of 10-HDA on human colon cancer cells, WiDr, as well as its effect on the growth of pathogenic bacterium. METHODS: The pro-inflammatory cytokines, receptor antagonist cytokine (IL-1ra) and nuclear factor-kappa B (NF-κB) in WiDr cells was analyzed by Enzyme-linked immunosorbent assay (ELISA) or western blot. The growth inhibition of 10-HDA on bacterium was evaluated by determination of minimal inhibitory concentrations (MIC) and minimal bactericide concentrations (MBC). RESULTS: The production of pro-inflammatory cytokines, Interleukin (IL)-8, IL-1ß and tumor necrosis factor-alpha (TNF-α) in WiDr cells was modulated by 10-HDA. IL-8 were dramatically declined by 10-HDA at 3 mM, while IL-1ß and TNF-α were significantly decreased. 10-HDA increased IL-1ra in a dose manner. NF-κB pathway is primarily in response to prototypical pro-inflammatory cytokines, and NF-κB was reduced after 10-HDA treatment. 10-HDA acted as potent bactericide against animal- or human-specific pathogens, including Staphylococcus aureus, Streptococcus alactolyticus, Staphylococcus intermedius B, Staphylococcus xylosus, Salmonella cholearasuis, Vibro parahaemolyticus and Escherichia coli (hemolytic). CONCLUSIONS: The current study showed that in vitro 10-HDA from RJ exhibited anti-inflammatory activity in WiDr cells, as well as anti-bacterial activity against animal pathogens. 10-HDA showed its potential as anti-imflammtory agent and bactericide to benefit human gastrointestinal tract.

Updates on the molecular epidemiology of Enterovirus D68 after installation of screening test among acute flaccid paralysis patients in Taiwan.

In respond to acute flaccid paralysis (AFP) in association with Enterovirus D68 (EV-D68) infection, Taiwan Centers for Disease Control began to screen EV-D68 infection among each AFP patient since July 2015 and detected the first case in August 2016. This article updated the molecular epidemiology trends of EV-D68 from the national surveillance data.

The functional property of royal jelly 10-hydroxy-2-decenoic acid as a melanogenesis inhibitor.

BACKGROUND: It has been reported that royal jelly would reduce melanin synthesis and inhibit the expression of melanogensis related proteins and genes. In this study, we evaluate the anti-melanogenic and depigmenting activity of 10-hydroxy-2-decenoic acid (10-HDA) from royal jelly of Apis mellifera. METHODS: In this study, we assesses the 10-HDA whitening activity in comparison with the changes in the intracellular tyrosinase activity, melanin content and melanin production related protein levles in B16F1 melanoma cells after treating with 10-HDA. Furthermore, the skin whitening effect was evaluated by applying a cream product containing with 0.5%, 1% and 2% of 10-HDA onto the skin of mice (C57BL/6 J) for 3 week to observe the effect of DL*-values. RESULTS: The results showed that 10-HDA inhibited the MITF protein expression (IC50 0.86 mM) in B16F1 melanoma cells. Western blot analysis revealed that 10-HDA inhibited the activity of tyrosinase and the expression of tyrosinase-related protein 1 (TRP-1), TRP-2, and microphthalmia-associated transcription factor (MITF) in B16F1 melanoma cells. In addition, the 10-HDA was applied on the skin of mice show significantly increased the average skin-whitening index (L value). CONCLUSIONS: The validation data indicated the potential of 10-HDA for use in suppressing skin pigmentation. The 10-HDA is proposed as a candidate to inhibit melanogenesis, thus it could be developed as cosmetics skin care products.

FAM20A binds to and regulates FAM20C localization.

Mutations in the Family with sequence similarity (FAM) 20 gene family are associated with mineralized tissue phenotypes in humans. Among these genes, FAM20A mutations are associated with Amelogenesis Imperfecta (AI) with gingival hyperplasia and nephrocalcinosis, while FAM20C mutations cause Raine syndrome, exhibiting bone and craniofacial/dental abnormalities. Although it has been demonstrated that Raine syndrome associated-FAM20C mutants prevented FAM20C kinase activity and secretion, overexpression of the catalytically inactive D478A FAM20C mutant was detected in both cell extracts and the media. This suggests that FAM20C secretion doesn't require its kinase activity, and that another molecule(s) may control the secretion. In this study, we found that extracellular FAM20C localization was increased when wild-type (WT), but not AI-forms of FAM20A was co-transfected. On the other hand, extracellular FAM20C was absent in the conditioned media of mouse embryonic fibroblasts (MEFs) derived from Fam20a knock-out (KO) mouse, while it was detected in the media from WT MEFs. We also showed that cells with the conditioned media of Fam20a WT MEFs mineralized, but those with the conditioned media of KO MEFs failed to mineralize in vitro. Our data thus demonstrate that FAM20A controls FAM20C localization that may assist in the extracellular function of FAM20C in mineralized tissues.

Morphological Variability and Distinct Protein Profiles of Cultured and Endosymbiotic Symbiodinium cells Isolated from Exaiptasia pulchella.

Symbiodinium is a dinoflagellate that plays an important role in the physiology of the symbiotic relationships of Cnidarians such as corals and sea anemones. However, it is very difficult to cultivate free-living dinoflagellates after being isolated from the host, as they are very sensitive to environmental changes. How these symbiont cells are supported by the host tissue is still unclear. This study investigated the characteristics of Symbiodinium cells, particularly with respect to the morphological variability and distinct protein profiles of both cultured and endosymbiotic Symbiodinium which were freshly isolated from Exaiptasia pulchella. The response of the cellular morphology of freshly isolated Symbiodinium cells kept under a 12 h L:12 h D cycle to different temperatures was measured. Cellular proliferation was investigated by measuring the growth pattern of Symbiodinium cells, the results of which indicated that the growth was significantly reduced in response to the extreme temperatures. Proteomic analysis of freshly isolated Symbiodinium cells revealed twelve novel proteins that putatively included transcription translation factors, photosystem proteins, and proteins associated with energy and lipid metabolism, as well as defense response. The results of this study will bring more understandings to the mechanisms governing the endosymbiotic relationship between the cnidarians and dinoflagellates.

Structure and antimicrobial activity relationship of royalisin, an antimicrobial peptide from royal jelly of Apis mellifera.

Royalisin is a 5.5-kDa antibacterial peptide isolated from the royal jelly of the honeybee (Apis mellifera). The antimicrobial activity of royalisin against fungi, Gram-positive and Gram-negative bacteria has been revealed. Compared with another insect antibacterial peptide, there is an extra stretch of 11 amino acid residues at the C-terminus of royalisin. In this study, a recombinant shortened form of royalisin named as royalisin-D, was constructed without the 11 amino acid residues at the C-terminal of royalisin and linked to the C-terminal of oleosin by an inteinS fragment. The recombinant protein was overexpressed in Escherichia coli, purified by artificial oil body system and subsequently released through self-splicing of inteinS induced by the changes of temperature. The antibacterial activity of royalisin-D was compared with royalisin via minimal inhibitory concentration (MIC) assay, minimal bactericidal concentration (MBC) assay, microbial adhesion to solvents (MATS) methods, and cell membrane permeability. Furthermore, the recombinant royalisin and royalisin-D have also been treated with the reducing agent of disulfide bonds, dithiothreitol (DTT), to investigate the importance of the intra-disulfide bond in royalisin. In our results, royalisin-D exhibited similar antimicrobial activity to royalisin. Royalisin and royalisin D lost their antimicrobial activities when the intra-disulfide bonds were reduced by DDT. The intra-disulfide bond plays a more important role than the extra stretch of 11 amino acid residues at the C-terminus of royalisin in terms of the antimicrobial properties of the native royalisin.