Self-perception of glycemic control among Japanese type 2 diabetic patients: accuracy of patient perception and characteristics of patients with misperception.

AIMS: Some diabetic patients, despite reporting a good perception of their glycemic control, actually show poor control and this misperception might well hinder successful diabetes management. This study aimed to assess patients' self-perception of glycemic control and to clarify factors associated with misperception of glycemic control status. METHODS: Baseline data from a hospital-based prospective cohort of 519 type 2 diabetic patients were analyzed. Self-perception of glycemic control and other items, including sociodemographic factors and blood test data, were determined from a self-administered questionnaire and medical records. Factors associated with misperception were examined by age group (elderly [aged ≥ 65 years] vs non-elderly [aged < 65 years]) using multiple logistic regression analysis. RESULTS: Among poorly controlled patients, misperception was higher in the elderly (glycated hemoglobin [HbA1c] 7.4-8.3, 55.1%; HbA1c >8.4, 44.8%) than in the non-elderly (HbA1c 7.4-8.3, 20.0%; HbA1c >8.4, 18.9%). The factors significantly associated with misperception were as follows: high lifestyle regimen adherence in both age groups (non-elderly group odds ratio [OR] 5.23; elderly group OR 5.15, respectively); high family support (OR = 7.32), failure to achieve blood pressure control (OR = 6.94) and having diabetic complications (OR = 0.06) among the non-elderly; and long duration of diabetes (OR = 4.06) among the elderly. CONCLUSIONS: For better management of diabetes, physicians should pay attention to the patient characteristics associated with misperception among uncontrolled diabetic patients, particularly among those who are elderly.

CD47 regulates the TGF-ß signaling pathway in osteoblasts and is distributed in Meckel's cartilage.

Previously, CD47 gene expression has been shown to increase during mandible development using a micro array technique. To determine the function of CD47 in osteoblasts, CD47 was silenced using siRNA in vitro. The TGF-ß1 and phosphorylated-Smad2 levels and transcription factor genes related to bone metabolism increased dose-dependently with CD47 silencing. Furthermore, we determined the distribution of CD47 in mouse embryonic E13 and E15 in vivo. The CD47-positive cells were localized in Meckel's cartilage and antenatal mandibular bone. These results suggest that TGF-ß1 signaling and mandible development might be regulated by CD47.

The distances between the facial and palatal papillae in the maxillary anterior dentition.

This study examined the distances between the facial and palatal papillae (DFPP) in the maxillary anterior dentition. A total of 360 filled interdental papillae in 72 study models were examined (Group A: 36 young patients, Group B: 36 adult patients). The DFPP was measured directly on alginate impressions. In Group A, the average DFPP was 1.27 ± 0.6, 1.53 ± 0.42, and 1.83 ± 0.56 mm between the central incisors (CI-CI), central and lateral incisors (CI-LI), and lateral incisor and canine (LI-CA), respectively. In Group B, the respective average DFPPs were 1.3 ± 0.83, 1.38 ± 0.68, and 1.62 ± 0.58 mm. Within the limits of the present study, the average distances between the facial and palatal papillae seem to be useful for setting the proximal contact position.

Inhibitory effects of low-energy pulsed ultrasonic stimulation on cell surface protein antigen C through heat shock proteins GroEL and DnaK in Streptococcus mutans.

This study concerns the use of low-energy pulsed ultrasound as nondestructive photodynamic antimicrobial therapy for controlling dental plaque. We examined the antibacterial and bactericidal effects of low-energy pulsed ultrasound on mutans streptococci and its inhibitory effects on bacterial cell adhesion of Streptococcus mutans. The results indicated weak antibacterial and bactericidal effects. However, ultrasonic stimulation for less than 20 min markedly decreased bacterial cell adhesion. To analyze the mechanism underlying the inhibitory effect, we examined cell surface protein antigen C (PAc) and glucosyltransferase I (GTF-I) expression in S. mutans. The levels of PAc gene and protein expression were markedly decreased by ultrasonic stimulation for 20 min. However, no change in GTF-I expression was observed. The expression of stress response heat shock proteins GroEL and DnaK was also examined. GroEL and DnaK levels were significantly decreased by ultrasonic stimulation, and the expression of the PAc protein was also diminished upon the addition of GroEL or DnaK inhibitors without ultrasonic stimulation. These observations suggest that the expression of the PAc protein in S. mutans may be dependent on heat shock proteins. Thus, low-energy pulsed ultrasound decreases bacterial adhesion by the inhibitory effect on the PAc protein and heat shock protein expression and may be useful as photodynamic antimicrobial chemotherapy in controlling dental plaque.

Traf2 interacts with Smad4 and regulates BMP signaling pathway in MC3T3-E1 osteoblasts.

Bone morphogenetic proteins (BMPs) play important roles in osteoblast differentiation and maturation. In mammals, the BMP-induced receptor-regulated Smads form complexes with Smad4. These complexes translocate and accumulate in the nucleus, where they regulate the transcription of various target genes. However, the function of Smad4 remains unclear. We performed a yeast two-hybrid screen using Smad4 as bait and a cDNA library derived from bone marrow, to indentify the proteins interacting with Smad4. cDNA clones for Tumor necrosis factor (TNF) receptor-associated factor 2 (Traf2) were identified, and the interaction between the endogenous proteins was confirmed in the mouse osteoblast cell line MC3T3-E1. To investigate the function of Traf2, we silenced it with siRNA. The level of BMP-2 protein in the medium, the expression levels of the Bmp2 gene and BMP-induced transcription factor genes, including Runx2, Dlx5, Msx2, and Sp7, and the phosphorylated-Smad1 protein level were increased in cells transfected with Traf2 siRNA. The nuclear accumulation of Smad1 increased with TNF-alpha stimulation for 30 min at Traf2 silencing. These results suggest that the TNF-alpha-stimulated nuclear accumulation of Smad1 may be dependent on Traf2. Thus, the interaction between Traf2 and Smad4 may play a role in the cross-talk between TNF-alpha and BMP signaling pathways.

The in vitro osteogenetic characteristics of primary osteoblastic cells from a rabbit calvarium.

Previously, we showed that recombinant human bone morphogenetic protein-2 (rhBMP-2) increased bone augmentation beyond the skeletal envelope within a titanium cap in a rabbit calvarium; many cuboidal osteoblastic cells were observed histologically. These results suggested that the new osteoblastic cells might have differentiated and matured via stimulation by rhBMP-2. To date, however, no studies have reported the characteristics of osteoblastic cells derived from adult rabbit calvarium, after addition of rhBMP-2. To determine the effects of rhBMP-2 on osteoblastic cells, we observed morphological characteristics and alkaline phosphatase activity of osteoblastic cells from an adult rabbit calvarium. The expression of proteins in the BMP signaling pathway and extracellular matrix were analyzed, and mineralized nodule formation was assessed. The alkaline phosphatase activity increased significantly after rhBMP-2 stimulation. The protein levels of phosphorylated-Smad1, Runx2, osteocalcin, osteopontin, and type I collagen were augmented by rhBMP-2 stimulation using Western blotting or ELISA; rhBMP-2 also stimulated mineralized nodule formation with alizarin red staining. The results suggest that primary osteoblastic cells derived from a rabbit calvarium have osteogenetic characteristics in vitro, underscoring the potential use of these cells as a model for studying bone formation. These cells may play an important role in in vivo bone augmentation in a rabbit experimental model.

Glycolytic enzyme GAPDH promotes peroxide stress signaling through multistep phosphorelay to a MAPK cascade.

Phosphorelay signaling of environmental stimuli by two-component systems is prevailing in bacteria and also utilized by fungi and plants. In the fission yeast Schizosaccharomyces pombe, peroxide stress signals are transmitted from the Mak2/3 sensor kinases to the Mpr1 histidine-containing phosphotransfer (HPt) protein and finally to the Mcs4 response regulator, which activates a MAP kinase cascade. Here we show that, unexpectedly, the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) physically associates with the Mcs4 response regulator and stress-responsive MAP kinase kinase kinases (MAPKKKs). In response to H2O2 stress, Cys-152 of the Tdh1 GAPDH is transiently oxidized, which enhances the association of Tdh1 with Mcs4. Furthermore, Tdh1 is essential for the interaction between the Mpr1 HPt protein and the Mcs4 response regulator and thus for phosphorelay signaling. These results demonstrate that the glycolytic enzyme GAPDH plays an essential role in the phosphorelay signaling, where its redox-sensitive cysteine residue may provide additional input signals.

Ubc9 promotes the stability of Smad4 and the nuclear accumulation of Smad1 in osteoblast-like Saos-2 cells.

Bone morphogenetic proteins (BMPs) play important roles in osteoblast differentiation and maturation. In mammals, the BMP-induced receptor-regulated Smads form complexes with Smad4. These complexes translocate and accumulate within the nucleus, where they regulate the transcription of various target genes. However, the function of Smad4 remains unclear. We performed a yeast two-hybrid screen using Smad4 as bait and a cDNA library derived from human bone marrow to identify the proteins interacting with Smad4. Two full-length cDNA clones for Ubc9 were identified, and the potential functions of Ubc9 were investigated. To determine the role of Ubc9 in the BMP signaling pathway, the endogenous transcription of Ubc9 in the human osteoblast cell line Saos-2 was silenced using siRNA. The expression of BMP-induced transcription factors, including Runx2, Dlx5, Msx2, and Osterix, was examined using real-time reverse transcription polymerase chain reaction (qRT-PCR), and the protein expression of Smad4, Smad1, phosphorylated Smad1, and BMP type I receptors was determined by Western blotting. The subcellular localization of Smad1 and Smad4 was observed using immunofluorescence staining after Ubc9 silencing. To determine whether Smad4 is sumoylated in vitro, recombinant Smad4 was purified and sumoylated Smad4 was visualized using Western blotting. The mRNA expression of various transcription factors was markedly inhibited after Ubc9 silencing. The protein levels of Smad4 and phosphorylated Smad1 decreased in a dose-dependent manner according to the amount of siRNA applied. Gene silencing also decreased the nuclear accumulation of Smad1 and Smad4. The sumoylation assay showed that sumoylated Smad4 is present and dependent on Ubc9 in vitro, which was confirmed by pretreatment with Senp2, a SUMO-protease. These results suggest that Ubc9 promotes the stability of sumoylated Smad4. Furthermore, the expression of key transcription factors, phosphorylated Smad1 protein, and the nuclear accumulation of Smad1 and Smad4 are inhibited by Ubc9 silencing. Thus, Ubc9 plays an important role in the up-regulation of the BMP signaling pathway.

Effects of low-intensity pulsed ultrasound on the differentiation of C2C12 cells.

Low-intensity pulsed ultrasound (LIPUS) is known to accelerate bone regeneration, but the precise cellular mechanism is still unclear. The purpose of this study was to determine the effect of LIPUS on the differentiation of pluripotent mesenchymal cell line C2C12. The cells were cultured in differentiation medium with or without the addition of LIPUS stimulation. The ultrasound signal consisted of 1.5 MHz at an intensity of 70 mW/cm2 for 20 min for all cultures. To verify the cell lineage after LIPUS stimulation, mRNA expression of cellular phenotype-specific markers characterizing osteoblasts (Runx2, Msx2, Dlx5, AJ18), chondroblasts (Sox9), myoblasts (MyoD), and adipocytes (C/EBP, PPARgamma) was studied using real-time polymerase chain reaction analysis. The protein expression of Runx2 and activated phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 mitogen-activated protein kinase (p38 MAPK) were performed using Western blotting. The mRNA expression of Runx2, Msx2, Dlx5, AJ18, and Sox9 was increased markedly by the LIPUS stimulation, whereas the expression of MyoD, C/EBP, and PPARgamma was drastically decreased. In the Western blot analysis, LIPUS stimulation increased Runx2 protein expression and phosphorylation of ERK1/2 and p38 MAPK. Our study demonstrated that LIPUS stimulation converts the differentiation pathway of C2C12 cells into the osteoblast and/or chondroblast lineage via activated phosphorylation of ERK1/2 and p38 MAPK.

Inhibitory effects of a super pulsed carbon dioxide laser at low energy density on periodontopathic bacteria and lipopolysaccharide in vitro.

OBJECTIVE AND BACKGROUND: Previous studies have described the effect of irradiation by a carbon dioxide (CO2) laser at high energy density on oral bacteria, and various side-effects have also been observed. However, no published studies have examined the effect of irradiation by a CO2 laser at low energy density on oral bacteria. The purpose of this study was to investigate the effects of super pulsed CO2 laser irradiation on periodontopathic bacteria and lipopolysaccharide (LPS). METHODS: Bacterial suspensions of two species of periodontopathic bacteria received laser irradiation at energy densities of 0-12.5 J/cm2. The suspensions were then spread over agar plates and incubated anaerobically. The bactericidal effects were evaluated based on colony formation. Samples of LPS were laser-irradiated at energy densities of 0-12.5 J/cm2. The biological activity was measured, and LPS was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: The irradiation at low energy densities of 7.5 and 12.5 J/cm2 killed more than 99.9 and 99.999% of Porphyromonas gingivalis and more than 99% of Actinobacillus actinomycetemcomitans was sterilized by the irradiation at 7.5 J/cm2. LPS biological activity was significantly decreased by laser irradiation at energy densities of more than 7.5 J/cm2 (p < 0.05), and the components of LPS analyzed by SDS-PAGE was diminished non-specifically. CONCLUSION: The results indicate that CO2 laser irradiation at low power is capable of bactericidal effect on periodontopathic bacteria and decreasing LPS activity.

Wsh3/Tea4 is a novel cell-end factor essential for bipolar distribution of Tea1 and protects cell polarity under environmental stress in S. pombe.

BACKGROUND: The fission yeast Schizosaccharomyces pombe has a cylindrical cell shape, for which growth is strictly limited to both ends, and serves as an excellent model system for genetic analysis of cell-polarity determination. Previous studies identified a cell-end marker protein, Tea1, that is transported by cytoplasmic microtubules to cell tips and recruits other cell-end factors, including the Dyrk-family Pom1 kinase. The deltatea1 mutant cells cannot grow in a bipolar fashion and show T-shaped morphology after heat shock. RESULTS: We identified Wsh3/Tea4 as a novel protein that interacts with Win1 MAP kinase kinase kinase (MAPKKK) of the stress-activated MAP kinase cascade. Wsh3 forms a complex with Tea1 and is transported to cell tips by growing microtubules. The deltawsh3 mutant shows monopolar growth with abnormal Tea1 aggregate at the non-growing cell end; this abnormal aggregate fails to recruit Pom1 kinase. Consistent with the observed interaction between Win1 and Wsh3, cells lacking Wsh3 or Tea1 show more severe cell-polarity defects under osmolarity and heat-stress stimuli that are known to activate the stress MAPK cascade. Furthermore, mutants of the stress MAPK also exhibit cell-polarity defects when exposed to the same stress. CONCLUSIONS: Wsh3/Tea4 is an essential component of the Tea1 cell-end complex. In addition to its role in bipolar growth during the normal cell cycle, the Wsh3-Tea1 complex, together with the stress-signaling MAPK cascade, contributes to cell-polarity maintenance under stress conditions.

Response of fission yeast to toxic cations involves cooperative action of the stress-activated protein kinase Spc1/Sty1 and the Hal4 protein kinase.

Stress-activated protein kinases (SAPKs), members of a mitogen-activated protein kinase (MAPK) subfamily, are highly conserved among eukaryotes. Studies of yeasts demonstrated that SAPKs play pivotal roles in survival responses to high osmolarity, oxidative stress, and heat shock. Here we report a novel physiological role of the fission yeast Spc1 SAPK in cellular resistance to certain cations, such as Na(+), Li(+), and Ca(2+). Strains lacking Spc1 or its activator, Wis1 MAPK kinase, are hypersensitive to these cations. Spc1 positively regulates expression of sod2(+) encoding a Na(+)/H(+) antiporter through Atf1 and other transcription factors. In addition, we have identified a novel Spc1-interacting protein, Hal4, which is highly homologous to the budding yeast Sat4/Hal4 protein kinase. Like its budding yeast counterpart, the fission yeast Hal4 kinase is essential for cellular resistance to Na(+), Li(+), and Ca(2+). The hal4-null phenotype is complemented by overexpression of the Trk1 potassium transporter or increased K(+) in the growth medium, suggesting that Hal4 promotes K(+) uptake, which consequently increases cellular resistance to other cations. Interestingly, the Spc1-Hal4 interaction appears to be required for cellular resistance to Ca(2+) but not Na(+) and Li(+). We propose that Spc1 SAPK and Hal4 kinase cooperatively function to protect cells from the toxic cations.