Alternative polyadenylation determines the functional landscape of inverted Alu repeats.

Inverted Alu repeats (IRAlus) are abundantly found in the transcriptome, especially in introns and 3' untranslated regions (UTRs). Yet, the biological significance of IRAlus embedded in 3' UTRs remains largely unknown. Here, we find that 3' UTR IRAlus silences genes involved in essential signaling pathways. We utilize J2 antibody to directly capture and map the double-stranded RNA structure of 3' UTR IRAlus in the transcriptome. Bioinformatic analysis reveals alternative polyadenylation as a major axis of IRAlus-mediated gene regulation. Notably, the expression of mouse double minute 2 (MDM2), an inhibitor of p53, is upregulated by the exclusion of IRAlus during UTR shortening, which is exploited to silence p53 during tumorigenesis. Moreover, the transcriptome-wide UTR lengthening in neural progenitor cells results in the global downregulation of genes associated with neurodegenerative diseases, including amyotrophic lateral sclerosis, via IRAlus inclusion. Our study establishes the functional landscape of 3' UTR IRAlus and its role in human pathophysiology.

Long-term effect of implant-abutment connection type on marginal bone loss and survival of dental implants.

PURPOSE: This study aimed to compare the long-term survival rate and peri-implant marginal bone loss between different types of dental implant-abutment connections. METHODS: Implants with external or internal abutment connections, which were fitted at Gangneung-Wonju National University Dental Hospital from November 2011 to December 2015 and followed up for >5 years, were retrospectively investigated. Cumulative survival rates were evaluated for >5 years, and peri-implant marginal bone loss was evaluated at 1- and 5-year follow-up examinations after functional loading. RESULTS: The 8-year cumulative survival rates were 93.3% and 90.7% in the external and internal connection types, respectively (P=0.353). The mean values of marginal bone loss were 1.23 mm (external) and 0.72 mm (internal) (P<0.001) after 1 year of loading, and 1.20 mm and 1.00 mm for external and internal abutment connections, respectively (P=0.137) after 5 years. Implant length (longer, P=0.018), smoking status (heavy, P=0.001), and prosthetic type (bridge, P=0.004) were associated with significantly greater marginal bone loss, and the use of screw-cement-retained prosthesis was significantly associated (P=0.027) with less marginal bone loss. CONCLUSIONS: There was no significant difference in the cumulative survival rate between implants with external and internal abutment connections. After 1 year of loading, marginal bone loss was greater around the implants with an external abutment connection. However, no significant difference between the external and internal connection groups was found after 5 years. Both types of abutment connections are viable treatment options for the reconstruction of partially edentulous ridges.

ZNF598 co-translationally titrates poly(GR) protein implicated in the pathogenesis of C9ORF72-associated ALS/FTD.

C9ORF72-derived dipeptide repeat proteins have emerged as the pathogenic cause of neurodegeneration in amyotrophic lateral sclerosis and frontotemporal dementia (C9-ALS/FTD). However, the mechanisms underlying their expression are not fully understood. Here, we demonstrate that ZNF598, the rate-limiting factor for ribosome-associated quality control (RQC), co-translationally titrates the expression of C9ORF72-derived poly(GR) protein. A Drosophila genetic screen identified key RQC factors as potent modifiers of poly(GR)-induced neurodegeneration. ZNF598 overexpression in human neuroblastoma cells inhibited the nuclear accumulation of poly(GR) protein and decreased its cytotoxicity, whereas ZNF598 deletion had opposing effects. Poly(GR)-encoding sequences in the reporter RNAs caused translational stalling and generated ribosome-associated translation products, sharing molecular signatures with canonical RQC substrates. Furthermore, ZNF598 and listerin 1, the RQC E3 ubiquitin-protein ligase, promoted poly(GR) degradation via the ubiquitin-proteasome pathway. An ALS-relevant ZNF598R69C mutant displayed loss-of-function effects on poly(GR) expression, as well as on general RQC. Moreover, RQC function was impaired in C9-ALS patient-derived neurons, whereas lentiviral overexpression of ZNF598 lowered their poly(GR) expression and suppressed proapoptotic caspase-3 activation. Taken together, we propose that an adaptive nature of the RQC-relevant ZNF598 activity allows the co-translational surveillance to cope with the atypical expression of pathogenic poly(GR) protein, thereby acquiring a neuroprotective function in C9-ALS/FTD.

The trinity of ribosome-associated quality control and stress signaling for proteostasis and neuronal physiology.

Translating ribosomes accompany co-translational regulation of nascent polypeptide chains, including subcellular targeting, protein folding, and covalent modifications. Ribosome-associated quality control (RQC) is a co-translational surveillance mechanism triggered by ribosomal collisions, an indication of atypical translation. The ribosome-associated E3 ligase ZNF598 ubiquitinates small subunit proteins at the stalled ribosomes. A series of RQC factors are then recruited to dissociate and triage aberrant translation intermediates. Regulatory ribosomal stalling may occur on endogenous transcripts for quality gene expression, whereas ribosomal collisions are more globally induced by ribotoxic stressors such as translation inhibitors, ribotoxins, and UV radiation. The latter are sensed by ribosome-associated kinases GCN2 and ZAKα, activating integrated stress response (ISR) and ribotoxic stress response (RSR), respectively. Hierarchical crosstalks among RQC, ISR, and RSR pathways are readily detectable since the collided ribosome is their common substrate for activation. Given the strong implications of RQC factors in neuronal physiology and neurological disorders, the interplay between RQC and ribosome-associated stress signaling may sustain proteostasis, adaptively determine cell fate, and contribute to neural pathogenesis. The elucidation of underlying molecular principles in relevant human diseases should thus provide unexplored therapeutic opportunities. [BMB Reports 2021; 54(9): 439-450].

Correlation analysis of gingival recession after orthodontic treatment in the anterior region: an evaluation of soft and hard tissues.

PURPOSE: The aim of this study was to investigate and identify the main causes of periodontal tissue change associated with labial gingival recession by examining the anterior region of patients who underwent orthodontic treatment. METHODS: In total, 45 patients who had undergone orthodontic treatment from January 2010 to December 2015 were included. Before and after the orthodontic treatment, sectioned images from 3-dimensional digital model scanning and cone-beam computed tomography images in the same region were superimposed to measure periodontal parameters. The initial labial gingival thickness (IGT) and the initial labial alveolar bone thickness (IBT) were measured at 4 mm below the cementoenamel junction (CEJ), and the change of the labial gingival margin was defined as the change of the distance from the CEJ to the gingival margin. Additionally, the jaw, tooth position, tooth inclination, tooth rotation, and history of orthognathic surgery were investigated to determine the various factors that could have affected anterior periodontal tissue changes. RESULTS: The mean IGT and IBT were 0.77±0.29 mm and 0.77±0.32 mm, respectively. The mean gingival recession was 0.14±0.57 mm. Tooth inclination had a significant association with gingival recession, and as tooth inclination increased labially, gingival recession increased by approximately 0.2 mm per 1°. CONCLUSIONS: In conclusion, the IGT, IBT, tooth position, tooth rotation, and history of orthognathic surgery did not affect labial gingival recession. However, tooth inclination showed a significant association with labial gingival recession of the anterior teeth after orthodontic treatment.

Combining virtual model and cone beam computed tomography to assess periodontal changes after anterior tooth movement.

BACKGROUND: Orthodontic force may affect not only periodontal ligaments, but also the alveoloar bone and the gingiva according to the type of tooth movements. The authors assessed changes in gingival thickness (GT) and alveolar bone thickness (ABT) after orthodontic treatment using a new method. METHODS: This study included 408 teeth (208 central incisors, 200 lateral incisors) from the upper and lower 4 anterior teeth of 52 patients who had completed orthodontic treatment. GT and ABT were measured using virtual casts fabricated from impressions and cone beam computed tomography (CBCT). Two sectioned images of every tooth axis were acquired by partitioning each tooth with a line connecting the midpoint of the incisal edge to the midpoint of the cementoenamel junction in the virtual models and the root apex in CBCT images. After superimposing the two sectioned images, GT and ABT were measured before and after orthodontic tooth movement. Correlations between GT and ABT before and after treatment, and changes in GT and ABT associated with sex, tooth arch, tooth position, orthognathic surgery, and tooth inclination and rotation were assessed. RESULTS: Before orthodontic treatment, GT and ABT were significantly correlated. Patients who underwent orthognathic surgery exhibited an increase in GT thickness compared with those who did not. ABT was significantly decreased in proclined teeth and in rotated teeth. CONCLUSIONS: GT and ABT can be affected by the nature of tooth movement and can be accurately assessed by comparing sectioned CBCT images and virtual models.

The prognosis of splinted restoration of the most-distal implants in the posterior region.

PURPOSE: The aim of this study was to compare the efficacies of two-implant splinting (2-IS) and single-implant restoration (1-IR) in the first and second molar regions over a mean functional loading period (FLP) of 40 months, and to propose the appropriate clinical considerations for the splinting technique. MATERIALS AND METHODS: The following clinical factors were examined in the 1-IR and 2-IS groups based on the total hospital records of the patients: sex, mean age, implant location, FLP, bone grafting, clinical crown-implant ratio, crown height space, and horizontal distance. The mechanical complications [i.e., screw loosening (SL), screw fracture, crown fracture, and repeated SL] and biological complications [i.e., peri-implant mucositis (PM) and peri-implantitis (PI)] were also evaluated for each patient. In analysis of two groups, the chi-square test and Student's t-test were used to identify the relationship between clinical factors and complication rates. The optimal cutoff value for the FLP based on complications was evaluated using receiver operating characteristics analysis. RESULTS: In total, 234 patients with 408 implants that had been placed during 2005 - 2014 were investigated. The incident rates of SL (P<.001), PM (P=.002), and PI (P=.046) differed significantly between the 1-IR and 2-IS groups. The FLP was the only meaningful clinical factor for mechanical and biological complication rates in 2-IS. CONCLUSION: The mechanical complication rates were lower for 2-IS than for 1-IR, while the biological complication rates were higher for 2-IS. FLP of 39.80 and 46.57 months were the reference follow-up periods for preventing biological and mechanical complications, respectively.

Mechanical and biological complication rates of the modified lateral-screw-retained implant prosthesis in the posterior region: an alternative to the conventional Implant prosthetic system.

PURPOSE: The modified lateral-screw-retained implant prosthesis (LSP) is designed to combine the advantages of screw- and cement-retained implant prostheses. This retrospective study evaluated the mechanical and biological complication rates of implant-supported single crowns (ISSCs) inserted with the modified LSP in the posterior region, and determined how these complication rates are affected by clinical factors. MATERIALS AND METHODS: Mechanical complications (i.e., lateral screw loosening [LSL], abutment screw loosening, lateral screw fracture, and ceramic fracture) and biological complications (i.e., peri-implant mucositis [PM] and peri-implantitis) were identified from the patients' treatment records, clinical photographs, periapical radiographs, panoramic radiographs, and clinical indices. The correlations between complication rates and the following clinical factors were determined: gender, age, position in the jaw, placement location, functional duration, clinical crown-to-implant length ratio, crown height space, and the use of a submerged or nonsubmerged placement procedure. RESULTS: Mechanical and biological complications were present in 25 of 73 ISSCs with the modified LSP. LSL (n=11) and PM (n=11) were the most common complications. The incidence of mechanical complications was significantly related to gender (P=.018). The other clinical factors were not significantly associated with mechanical and biological complication rates. CONCLUSION: Within the limitations of this study, the incidence of mechanical and biological complications in the posterior region was similar for both modified LSP and conventional implant prosthetic systems. In addition, the modified LSP is amenable to maintenance care, which facilitates the prevention and treatment of mechanical and biological complications.

Mechanical Complication Rates and Optimal Horizontal Distance of the Most Distally Positioned Implant-Supported Single Crowns in the Posterior Region: A Study with a Mean Follow-Up of 3 Years.

PURPOSE: The purpose of this retrospective study was to determine the 3-year mechanical complication rates of the most distally positioned implant-supported single crowns (ISSCs) in the posterior region, and how these complication rates are affected by several clinical factors (i.e., gender, mean age, horizontal distance [HD], position in the jaw, placement location, duration of functional loading, clinical crown-to-implant length ratio [C/I ratio], and crown height space of the implant). MATERIALS AND METHODS: The mechanical complications (i.e., abutment screw loosening [ASL], abutment screw fracture [ASF], and ceramic fracture [CF]) associated with the implants were identified by examining the patients' treatment records, clinical photographs, and intraoral periapical radiographs. Statistical analyses were performed using chi-square and Student's t-test to identify the relationship between various clinical factors and the mechanical complication rates. Receiver operating characteristics analysis was conducted to determine the optimal cut-off value for the HD between the most distally positioned ISSCs and the mesially adjacent natural tooth beyond which complications occur. RESULTS: The study inclusion criteria were met by 183 patients who had undergone implant surgery in the period 2004 to 2011, involving a total of 221 implant treatments. Mechanical complications were present in 40 (18.1%) of the 221 investigated ISSCs. ASL was the most common complication (n = 28, 12.7%), followed in order by CF (n = 9, 4.1%) and ASF (n = 3, 1.4%). Repeated ASL and CF occurred in four (1.8%) and two (0.9%) implants, respectively. The mechanical complication rates differed significantly between implants with different HDs (p = 0.009) and clinical C/I ratios (p = 0.019); however, there was no significant association between the other clinical factors and the mechanical complication rates. CONCLUSION: Within the limitations of this study, it appears that the incidence of mechanical complications is higher for the most distally positioned ISSCs in the posterior region than for those positioned at other sites. Furthermore, since the rate of mechanical complications increases with increasing HD, an HD of 3.7 mm or less is recommended.

Synergistic Effects of a Calcium Phosphate/Fibronectin Coating on the Adhesion of Periodontal Ligament Stem Cells Onto Decellularized Dental Root Surfaces.

This study aimed to enhance the attachment of periodontal ligament stem cells (PDLSCs) onto the decellularized dental root surface using surface coating with fibronectin and/or calcium phosphate (CaP) and to evaluate the activity of PDLSCs attached to a coated dental root surface following tooth replantation. PDLSCs were isolated from five dogs, and the other dental roots were used as a scaffold for carrying PDLSCs and then assigned to one of four groups according to whether their surface was coated with CaP, fibronectin, CaP/fibronectin, or left uncoated (control). Fibronectin increased the adhesion of PDLSCs onto dental root surfaces compared to both the control and CaP-coated groups, and simultaneous surface coating with CaP and fibronectin significantly accelerated and increased PDLSC adhesion compared to the fibronectin-only group. On in vivo tooth replantation, functionally oriented periodontal new attachment was observed on the CaP/fibronectin-coated dental roots to which autologous PDLSCs had adhered, while in the control condition, dental root replantation was associated only with root resorption and ankylosis along the entire root length. CaP and fibronectin synergistically enhanced the attachment of PDLSCs onto dental root surfaces, and autologous PDLSCs could produce de novo periodontal new attachment in an experimental in vivo model.

The DOUBLETIME protein kinase regulates phosphorylation of the Drosophila PDP1epsilon.

Reversible phosphorylation of clock proteins plays an important role in circadian timekeeping as it is a key post-translational mechanism that regulates the activity, stability and subcellular localization of core clock proteins. The kinase DOUBLETIME (DBT), a Drosophila ortholog of mammalian casein kinase Iepsilon, regulates circadian phosphorylation of two essential clock proteins, PERIOD and dCLOCK. We present evidence that Par Domain Protein 1epsilon (PDP1epsilon), a transcription factor and mediator of clock output in Drosophila, is phosphorylated in vivo and in cultured cells by DBT activity. We also demonstrate that DBT interacts with PDP1epsilon and promotes its degradation by the ubiquitin-proteasome pathway in cultured cells. In addition, PDP1epsilon nuclear localization is decreased by dbt RNA interference in S2 cell system. These results suggest that DBT regulates phosphorylation, stability and localization of PDP1epsilon, and that it has multiple targets in the Drosophila circadian system.

Changes in FDG Tumor Uptake during and after Fractionated Radiation Therapy in a Rodent Tumor Xenograft.

OBJECTIVE: The uptake of FDG was measured before, during, and after fractionated radiation in order to evaluate the potential of FDG-PET imaging as an indicator of tumor response.METHODS: The study was performed with nude rats bearing the human neuroblastoma BE(2)C tumor xenografts. Tumors were irradiated with 10 fractions of 2 Gy using a 320 kV(p) X-ray unit. Following a baseline FDG-PET scan, repeat scans were performed weekly until animal sacrifice. The rodents were given up to 10 FDG-PET scans, over a period of up to 75 days posttreatment.RESULTS AND CONCLUSIONS: Neither, the average and maximum activity/cc of FDG tumor uptake, nor the respective standardized uptake values (SUV), correlated with tumor response. Instead, the total FDG uptake (defined as the product of the average FDG activity/cc with the tumor volume) correlated better with tumor response.