Meta-analysis of the transcriptome identifies aberrant RNA processing as common feature of aging in multiple species.

Aging is accompanied by the gradual deregulation of the transcriptome. However, whether age-dependent changes in the transcriptome are evolutionarily conserved or diverged remains largely unexplored. Here, we performed a meta-analysis examining the age-dependent changes in the transcriptome using publicly available datasets of 11 representative metazoans, ranging from Caenorhabditis elegans to humans. To identify the transcriptomic changes associated with aging, we analyzed various aspects of the transcriptome, including genome composition, RNA processing, and functional consequences. The use of introns and novel splice sites tended to increase with age, particularly in the brain. In addition, our analysis suggests that the age-dependent accumulation of premature termination codon-containing transcripts is a common feature of aging across multiple animal species. Using C. elegans as a test model, we showed that several splicing factors that are evolutionarily conserved and age-dependently downregulated were required to maintain a normal lifespan. Thus, aberrant RNA processing appears to be associated with aging and a short lifespan in various species.

Immunosenescence in Caenorhabditis elegans.

Immunosenescence is an age-dependent decline in immune functions and hallmark of aging in diverse species, ranging from invertebrates to mammals. However, identifying the factors responsible for immunosenescence is challenging because of the complexity of immune systems and aging in mammals. The roundworm Caenorhabditis elegans is suitable for understanding immunosenescence because of its simple immune system and rapid aging process. In this review, we discuss the advances in our understanding of immunosenescence in C. elegans. PMK-1/p38 mitogen-activated protein kinase (MAPK), SKN-1/NRF, and ZIP-10/bZIP transcription factor regulate immunosenescence through p38 MAPK and insulin/IGF-1 signaling pathways. Because these factors and pathways are evolutionarily conserved, the findings discussed in this review may help understand the mechanisms underlying immunosenescence and develop new treatment therapy for immunosenescence in humans.

Double-stranded RNA induction asa potential dynamic biomarkerfor DNA-demethylating agents.

Hypomethylating agents (HMAs), such as azacitidine and decitabine, induce cancer cell death by demethylating DNAs to promote the expression of tumor-suppressor genes. HMAs also reactivate the transcription of endogenous double-stranded RNAs (dsRNAs) that trigger the innate immune response and subsequent apoptosis via viral mimicry. However, the expression patterns of endogenous dsRNAs and their relevance in the efficacy of HMAs remain largely uninvestigated. Here, we employ amidine-conjugated spiropyran (Am-SP) to examine the dynamic expression pattern of total dsRNAs regulated by HMAs. By analyzing the bone-marrow aspirates of myelodysplastic syndrome or acute myeloid leukemia patients who received the HMAs, we find a dramatic increase in total dsRNA levels upon treatment only in patients who later benefited from the therapy. We further apply our approach in solid tumor cell lines and show that the degree of dsRNA induction correlates with the effectiveness of decitabine in most cases. Notably, when dsRNA induction is accompanied by increased expression of nc886 RNA, decitabine becomes ineffective. Collectively, our study establishes the potential application of monitoring the total dsRNA levels by a small molecule as an analytical method and a dynamic marker to predict the clinical outcome of the HMA therapy.

MON-2, a Golgi protein, promotes longevity by upregulating autophagy through mediating inter-organelle communications.

The Golgi apparatus regulates the process of modification and subcellular localization of macromolecules, including proteins and lipids. Aberrant protein sorting caused by defects in the Golgi leads to various diseases in mammals. However, the role of the Golgi apparatus in organismal longevity remained largely unknown. By employing a quantitative proteomic approach, we demonstrated that MON-2, an evolutionarily conserved Arf-GEF protein implicated in Golgi-to-endosome trafficking, promotes longevity via upregulating macroautophagy/autophagy in C. elegans. Our data using cultured mammalian cells indicate that MON2 translocates from the Golgi to the endosome under starvation conditions, subsequently increasing autophagic flux by binding LGG-1/GABARAPL2. Thus, Golgi-to-endosome trafficking appears to be an evolutionarily conserved process for the upregulation of autophagy, which contributes to organismal longevity.

MON-2, a Golgi protein, mediates autophagy-dependent longevity in Caenorhabditis elegans.

The Golgi apparatus plays a central role in trafficking cargoes such as proteins and lipids. Defects in the Golgi apparatus lead to various diseases, but its role in organismal longevity is largely unknown. Using a quantitative proteomic approach, we found that a Golgi protein, MON-2, was up-regulated in long-lived Caenorhabditis elegans mutants with mitochondrial respiration defects and was required for their longevity. Similarly, we showed that DOP1/PAD-1, which acts with MON-2 to traffic macromolecules between the Golgi and endosome, contributed to the longevity of respiration mutants. Furthermore, we demonstrated that MON-2 was required for up-regulation of autophagy, a longevity-associated recycling process, by activating the Atg8 ortholog GABARAP/LGG-1 in C. elegans. Consistently, we showed that mammalian MON2 activated GABARAPL2 through physical interaction, which increased autophagic flux in mammalian cells. Thus, the evolutionarily conserved role of MON2 in trafficking between the Golgi and endosome is an integral part of autophagy-mediated longevity.

Age-dependent upregulation of Y RNAs in Caenorhabditis elegans.

Y RNA is a conserved small non-coding RNA whose functions in aging remain unknown. Here, we sought to determine the role of C. elegans Y RNA homologs, CeY RNA (CeY) and stem-bulge RNAs (sbRNAs), in aging. We found that the levels of CeY and sbRNAs generally increased during aging. We showed that CeY was downregulated by oxidative and thermal stresses, whereas several sbRNAs were upregulated by oxidative stress. We did not observe lifespan phenotypes by mutations in CeY-coding yrn-1. Future research under various genetic and environmental conditions is required to further evaluate the role of Y RNA in C. elegans aging.

Combinatorial Approach Using Caenorhabditis elegans and Mammalian Systems for Aging Research.

Aging is associated with functional and structural declines in organisms over time. Organisms as diverse as the nematode Caenorhabditis elegans and mammals share signaling pathways that regulate aging and lifespan. In this review, we discuss recent combinatorial approach to aging research employing C. elegans and mammalian systems that have contributed to our understanding of evolutionarily conserved aging-regulating pathways. The topics covered here include insulin/IGF-1, mechanistic target of rapamycin (mTOR), and sirtuin signaling pathways; dietary restriction; autophagy; mitochondria; and the nervous system. A combinatorial approach employing high-throughput, rapid C. elegans systems, and human model mammalian systems is likely to continue providing mechanistic insights into aging biology and will help develop therapeutics against age-associated disorders.

Different bone regeneration patterns in periimplant circumferential gap defects grafted with two types of osteoconductive biomaterial.

This study aimed to determine healing patterns in periimplant gap defect grafted with demineralized bovine bone mineral (DBBM) and porous titanium granules (PTG), which are known to induce a minimal tissue reaction and to undergo minimal biodegradation in healing process. Experiments were performed using a standardized periimplant gap-defect model in dogs with two observational periods: 4 and 8 weeks. Circumferential defects were surgically induced around dental implants on unilateral mandibles in five dogs, and collagen barrier membranes were placed over the DBBM and PTG grafts at two experimental sites and over a nongrafted site. Four weeks later, the same procedures were performed on the contralateral mandible, and the animals allowed to heal for a further 4 weeks, after which they were sacrificed and their mandibles with graft/control sites harvested for histologic evaluation. Both types of grafted biomaterials significantly enhanced the defect fill with newly formed bone, but the bone-to-implant contact (BIC) was significantly increased only at sites that had been grafted with DBBM. The two experimental sites exhibited different healing patterns, with new bone formation being observed on the surface of the DBBM particles throughout the defect, while there was no de novo bone formation on the PTG surface, but rather appositional bone growth from the base and lateral walls of the defect. It has been suggested that gap-defect filling with DBBM around dental implants may enhance both BIC and defect fill; however, the present findings show that defect grafting with PTG enhances only defect fill and not BIC. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1202-1209, 2016.

Simultaneous placement of an interpositional free gingival graft with nonsubmerged implant placement.

PURPOSE: Various surgical techniques target achieving adequate keratinized tissue around dental implants; however, these techniques are usually performed before implant placement or upon the exposure of submerged implants. The aim of this case report is to describe a simultaneous placement of an interpositional free gingival graft (iFGG) with that of nonsubmerged implants in a patient lacking keratinized tissue and to assess the long-term outcome of this grafted gingiva. METHODS: A wedge-shaped free gingnival graft (FGG), including an epithelium-connective tissue (E-C) portion and a connective-tissue-only (CT) portion, was harvested from the palate. The CT portion was inserted under the buccal flap, and the E-C portion was secured tightly around the implants and to the lingual flap. RESULTS: At the 8-year follow-up, the gingival graft remained firmly attached and was well maintained, with no conspicuous shrinkage or reported discomfort during oral hygiene procedures. The use of an iFGG at a nonsubmerged implant placement minimizes the required number of surgical steps and patient discomfort while providing adequate buccal keratinized tissue. CONCLUSIONS: Therefore, the technique could be considered an alternative method in increasing the keratinized tissue for cases that have a minimal amount of keratinized tissue.

Cumulative survival rate of Astra Tech implants: a retrospective analysis.

PURPOSE: The aim of this study was to analyze the short-term predictability and reliability of Astra Tech implants according to the demographical distribution of patients and condition of implant sites and location of implants. METHODS: Among patients treated with Astra Tech implant (Astra Tech AB) in the Department of Periodontology at the Dental Hospital of Yonsei University of College of Dentisry and K Dental Clinic from May 2004 to March 2009, 195 implants in 98 patients which had been restored more than 6 months ago were reviewed in this study. Following data were reviewed from patient charts and implants success rate was examined: 1) patient type and implant location, 2) bone status at the implant site, 3) diameter and length of the placed implants, 4) presence or absence of bone augmentation and types of the augmentation. RESULTS: THE RESULTS FROM THIS STUDY ARE AS FOLLOWS: 1) most implants were placed in the molar area, especially 1st molar area of maxilla, 2) most implants were placed at D2 and D3 bone type, 3) most implants were placed in areas of B and C bone quantity, 4) autogenous and alloplastic bone graft and artificial membrane were used for placement of 74 implants. CONCLUSIONS: Short-term survival rate of Astra Tech implants was 100%.

Spontaneous healing capacity of rabbit cranial defects of various sizes.

PURPOSE: This study evaluated the spontaneous healing capacity of surgically produced cranial defects in rabbits with different healing periods in order to determine the critical size defect (CSD) of the rabbit cranium. METHODS: Thirty-two New Zealand white rabbits were used in this study. Defects of three sizes (6, 8, and 11 mm) were created in each of 16 randomly selected rabbits, and 15-mm defects were created individually in another 16 rabbits. The defects were analyzed using radiography, histologic analysis, and histometric analysis after the animal was sacrificed at 2, 4, 8, or 12 weeks postoperatively. Four samples were analyzed for each size of defect and each healing period. RESULTS: The radiographic findings indicated that defect filling gradually increased over time and that smaller defects were covered with a greater amount of radiopaque substance. Bony islands were observed at 8 weeks at the center of the defect in both histologic sections and radiographs. Histometrical values show that it was impossible to determine the precise CSD of the rabbit cranium. However, the innate healing capacity that originates from the defect margin was found to be constant regardless of the defect size. CONCLUSIONS: The results obtained for the spontaneous healing capacity of rabbit cranial defects over time and the underlying factors may provide useful guidelines for the development of a rabbit cranial model for in vivo investigations of new bone materials.

Osteoconductive effects of calcium phosphate glass cement grafts in rabbit calvarial defects.

Calcium phosphate glass (CPG) is well-documented alloplastic bone graft material. The objective of this study was to evaluate the osteoconductive effect of newly developed calcium phosphate glass cement (CPGC) in rabbit calvarial defects. Three circular defects (8 mm diameter) were created on the rabbit calvarium. One defect was filled with biphasic calcium phosphate (BCP group) and one defect was filled with CPGC (CPGC group). The remaining defect was not filled as the control. Histologic and histometric analysis were performed at four and eight weeks following the implantation of materials. One-way ANOVA method was used to evaluated the significance between three groups (p < 0.05). The CPGC group did not show a statistical difference in new bone area compared with the control at all healing periods, but the bone formation rate of CPGC seemed to increase between four and eight weeks. This suggests the bone formation rate of CPGC is initially slow, but increases at a specific time, showing the possibility of greater bone formation with time. The resorption rate of CPGC was greater than BCP. Within the limits of this study, CPGC demonstrated good space maintaining capacity and had an osteoconductive effect, suggesting it could be successfully used to improve bone formation capacity.