Ablation of XP-V gene causes adipose tissue senescence and metabolic abnormalities

Obesity and the metabolic syndrome have evolved to be major health issues throughout the world. Whether loss of genome integrity contributes to this epidemic is an open question. DNA polymerase η (pol η), encoded by the xeroderma pigmentosum (XP-V) gene, plays an essential role in preventing cutaneous cancer caused by UV radiation-induced DNA damage. Herein, we demonstrate that pol η deficiency in mice (pol η^−/−) causes obesity with visceral fat accumulation, hepatic steatosis, hyperleptinemia, hyperinsulinemia, and glucose intolerance. In comparison to WT mice, adipose tissue from pol η^−/− mice exhibits increased DNA damage and a greater DNA damage response, indicated by up-regulation and/or phosphorylation of ataxia telangiectasia mutated (ATM), phosphorylated H₂AX (γH₂AX), and poly[ADP-ribose] polymerase 1 (PARP-1). Concomitantly, increased cellular senescence in the adipose tissue from pol η^−/− mice was observed and measured by up-regulation of senescence markers, including p53, p16^Ink4a, p21, senescence-associated (SA) β-gal activity, and SA secretion of proinflammatory cytokines interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) as early as 4 wk of age. Treatment of pol η^−/− mice with a p53 inhibitor, pifithrin-α, reduced adipocyte senescence and attenuated the metabolic abnormalities. Furthermore, elevation of adipocyte DNA damage with a high-fat diet or sodium arsenite exacerbated adipocyte senescence and metabolic abnormalities in pol η^−/− mice. In contrast, reduction of adipose DNA damage with N-acetylcysteine or metformin ameliorated cellular senescence and metabolic abnormalities. These studies indicate that elevated DNA damage is a root cause of adipocyte senescence, which plays a determining role in the development of obesity and insulin resistance.The human genome is constantly challenged by exogenous and endogenous DNA damaging agents. To ensure genome integrity, human cells have faithful DNA replication machinery and DNA repair systems that are coordinated by DNA damage response networks. In response to different extents or types of DNA lesions, the DNA damage response activates appropriate cellular responses, including transient or permanent (senescence) cell cycle arrest, or apoptosis, to minimize the detrimental effects of DNA lesions (1). Reduction or deficiency in DNA repair/replication enzyme activity is well documented to increase vulnerability for the development of cancer, neurodegenerative diseases, and aging (2). In addition, defective DNA repair enzymes are associated with the metabolic symptom; for example, DNA glycosylase (Neil1)- and OGG1-deficient mice are obese (3–5), and nucleotide excision repair protein ERCC1-XPF deficiency causes lipodystrophy (6). Furthermore, DNA damage response protein ataxia telangiectasia mutated (ATM) suppresses JNK activity through p53 signaling and mediates an antioxidant action that has been suggested to be relevant to the metabolic syndrome (7). Nucleotide excision repair XP-A protein may affect metabolism by altering mitochondrial function (8). To date, the mechanisms that link genome instability and metabolic dysregulation have not been elucidated.DNA polymerase η (pol η) is a specialized lesion bypass polymerase that faithfully replicates across UV-induced cyclobutane pyrimidine dimers (9) to rescue stalled DNA replication forks from potential breakages and mutations. Defects in the gene encoding pol η produce a variant form of the autosomal recessive disease xeroderma pigmentosum (XP-V) (9). Patients with XP-V are highly sensitive to sunlight and prone to cutaneous cancer (9). In addition to skin, pol η is expressed in most tissues (10). The expression of pol η correlates with the effectiveness of anticancer therapeutic agents that exert their activity by introducing DNA lesions that block the progression of DNA replication (11). In addition to exogenous introduced DNA lesions, pol η contributes to genomic stability during unperturbed DNA replication (12) and replicates across reactive oxygen species (ROS)-induced oxidative DNA lesions 8-oxoG (7,8-dihydro-8-oxoguanine), thymine glycol, and lipid peroxidation DNA adducts generated during endogenous metabolic processes (13, 14). ROS generated from endogenous metabolism or exogenous sources has been associated with cancer, aging, and the metabolic syndrome.Therefore, the initial hypothesis for our studies was that pol η contributes to reduce tumorigenesis by managing oxidative DNA lesions in other organs. However, in a 2-y study, no increase in the incidence of tumors of any type was found with pol η KO (pol η^−/−) mice (15). Instead, we found pol η^−/− mice develop metabolic abnormalities that induce obesity.     

Transpositioned flap vestibuloplasty combined with implant surgery in the severely resorbed atrophic edentulous ridge

The use of transpositioned flap (lipswitch) vestibuloplasty combined with implant surgery in patients with severely resorbed atrophic edentulous ridges is reviewed. The cases of 17 patients with severely resorbed atrophic edentulous ridges at the mandible undergoing implant rehabilitation were reviewed. Lipswitch vestibuloplasty was followed immediately by the implant surgery. Postoperative follow-up consisted of clinical and radiographic examinations. Seventeen patients with atrophic ridges (12 class II and 5 class III) each had 2 implant fixtures placed in the mandible as abutments for a clip and bar overdenture. The average time of follow-up was 6 years. Before surgery, all patients had severely atrophic ridges with a compromised shallow vestibule of varying degrees. Satisfactory results were observed in regard to the immediate and long-term morphology of the vestibule, the health of the peri-implant tissue, the stability of implant fixtures, and the functionality of the prostheses. The lipswitch vestibuloplasty offers a safe and convenient method of surgical access for implant fixture installation, with the advantage of rebuilding the vestibule of a compromised atrophic ridge in the anterior mandible.     

Resinous perforation-repair materials inhibit the growth,attachment, and proliferation of human gingival fibroblasts

The choice of repair material is one of the important factors in the prognosis of the endodontically treated tooth with a perforation defect. The cytotoxicity of perforation-repair materials must be investigated to ensure a safe biological response. The aim of this in vitro study was to evaluate the effect of resin-modified, glass-ionomer cement, compomer, and resin on human-gingival fibroblasts. Human gingival fibroblasts from crown lengthening surgery were cultured by using an explant technique with the consent of the patient. Cytotoxicity was judged by using an assay of tetrazolium bromide reduction. The results showed that resin-modified, glass-ionomer cement Fuji II LC, compomer Compoglass, and resin SpectrumTPH (TPH) were cytotoxic to primary human gingival fibroblast cultures by inhibiting cell growth and proliferation. TPH alone had an effect on cell attachment. It was found that TPH was the most cytotoxic repair material among those tested in all cultures. The toxicity decreased in the order of TPH>FLC>CG.     

Lactate dehydrogenase leakage of hepatocytes with AH26 and AH Plus sealer treatments

Numerous root canals filling materials are available in the field of dentistry, based on various formulas that contain a variety of different and partly mutagenic components, such as epoxy resin sealers, Ca(OH)2-based materials, and zinc oxide-eugenol cements. AH Plus root canal sealer will not release formaldehyde according to the manufacturer, although AH26 does. The purpose of this study was to analyze the leakage of lactate dehydrogenase (LDH) from rat hepatocytes after treatment with AH26 and AH Plus root canal sealers in vitro. Hepatocytes from male Sprague-Dawley rats were used to test the cytotoxicity of AH26 and AH Plus. The root canal sealers were mixed and then dissolved in the dimethyl sulfoxide to final concentrations of 0.01%, 0.04%, and 0.1% (wt/vol), with a dimethyl sulfoxide concentration of < 0.05%. Dosage-dependent and time-dependent lactate dehydrogenase leakage values were measured and tested by one-way ANOVA. The results showed that both AH26 and AH Plus are toxic to rat hepatocytes. At a low concentration, AH26 had a higher toxicity than AH Plus to rat hepatocytes.     

Stimulation of cells derived from nifedipine-induced gingival overgrowth with Porphyromonas gingivalis, lipopolysaccharide, and interleukin-1beta

The purpose of this study was to clarify the main contributory factor of nifedipine-induced gingival overgrowth either by Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) or interleukin-1beta (IL-1beta). Human gingival fibroblasts from healthy tissues and nifedipine-induced gingival overgrowth tissues were stimulated with nifedipine, IL-1beta, Escherichia coli lipopolysaccharide (Ec-LPS), and Pg-LPS, and the gene expressions were analyzed by RT-PCR. Analysis of the data showed no strong evidence of a synergistic effect of nifedipine and Pg-LPS on IL-6, connective tissue growth factor (CTGF), and type 1 collagen gene expression of either healthy cells or nifedipine-induced gingival overgrowth cells. Among the three stimulants--IL-1beta, Pg-LPS, and Ec-LPS--androgen receptor and IL-6 gene expressions in both the healthy and nifedipine-induced gingival overgrowth groups were strongly up-regulated by the presence of IL-1beta only. Furthermore, the responses to IL-1beta in the nifedipine-induced gingival overgrowth group were stronger than those of the healthy group. It can be concluded that IL-1beta is an important mediator responsible for the higher IL-6 and androgen receptor expression of nifedipine-induced gingival overgrowth cells.     

Long-term effect of xylitol gum use on mutans streptococci in adults

Many studies have shown the effects of chewing xylitol gum on mutans streptococci (MS) over short- and long-term periods in children; however, few studies have addressed long-term periods in adults. The objective of this investigation was to examine for 6 months the effects of chewing xylitol gum on MS in saliva and plaque in 127 adults (mean age 28.0 years). The participants were assigned to three groups according to gum type, in part taking preference for flavor into account and in part at random: xylitol (XYL), maltitol (MAL) and control (CR); 33, 34 and 27 subjects in each group, respectively, completed the trial. Daily gum use of the XYL and MAL groups was 7.9 and 7.1 g, respectively. MS levels, which declined significantly in saliva (p < 0.05) and plaque (p < 0.001) in the XYL group after 6 months, exhibited a significant increase in plaque in the MAL group (p < 0.001). Differences in relative changes of MS levels in plaque during the experimental period were significant between the XYL group and the CR (p < 0.05) and MAL groups (p < 0.001). Differences in relative change of amount of plaque during the experimental period were not statistically significant between the groups. The present study demonstrated that chewing xylitol gum for 6 months continued to inhibit the growth of mutans streptococci in adults.     

Reconstruction of the severely resorbed atrophic edentulous ridge of the maxilla and mandible for implant rehabilitation: report of a case

We describe a case with a severely resorbed atrophic edentulous ridge in both the maxilla and mandible. The maxilla was reconstructed using a sinus-lifting procedure and onlay bone graft. The mandible was reconstructed by anterior osteotomy with an interpositional sandwich iliac bone graft at the symphysis area, subperiosteally with iliac bone chips mixed with hydroxylapatite bilaterally at the posterior atrophic ridge, and vestibuloplasty with a split thickness skin graft (STSG). After full-mouth implant rehabilitation, a 5-year follow-up of this case shows a satisfactory result.     

Examination of the signal transduction pathways leading to upregulation of tissue type plasminogen activator by Porphyromonas endodontalis in human pulp cells

AIM: To investigate the tissue type plasminogen activator (t-PA) activity in human pulp cells stimulated with Porphyromonas endodontalis (P. endodontalis) in the absence or presence of p38 inhibitor SB203580, mitogen-activated protein kinase kinase (MEK) inhibitor U0126 and phosphatidylinositaol 3-kinase (PI3K) inhibitor LY294002. METHODOLOGY: The supernatants of P. endodontalis were used to evaluate t-PA activity in human pulp cells using casein zymography and enzyme-linked immunosorbent assay (ELISA). Furthermore, to search for possible signal transduction pathways, SB203580, U0126 and LY294002 were added to test how they modulated the t-PA activity. RESULTS: The main casein secreted by human pulp cells migrated at 70 kDa and represented t-PA. Secretion of t-PA was found to be stimulated with P. endodontalis during 2-day cultured period (P < 0.05). From the results of casein zymography and ELISA, SB203580 and U0126 significantly reduced the P. endodontalis stimulated t-PA production respectively (P < 0.05). However, LY294002 lacked the ability to change the P. endodontalis stimulated t-PA production (P > 0.05). CONCLUSIONS: Porphyromonas endodontalis enhances t-PA production in human pulp cells, and the signal transduction pathways p38 and MEK are involved in the inhibition of t-PA.