OBJECTIVE: To investigate the stem cell-like characteristics of human periodontal ligament (PDL) stromal cells outgrown from orthodontically extracted premolars and to evaluate the potential for myogenic differentiation. METHODS: PDL stromal cells were obtained from extracted premolars by using the outgrowth method. Cell morphological features, self-replication capability, and the presence of cell-surface markers, along with osteogenic, adipogenic, and chondrogenic differentiation, were confirmed. In addition, myogenic differentiation was induced by the use of 5-aza-2'-deoxycytidine (5-Aza) for DNA demethylation. RESULTS: PDL stromal cells showed growth patterns and morphological features similar to those of fibroblasts. In contrast, the proliferation rates of premolar PDL stromal cells were similar to those of bone marrow and adipogenic stem cells. PDL stromal cells expressed surface markers of human mesenchymal stem cells (i.e., CD90 and CD105), but not those of hematopoietic stem cells (i.e., CD31 and CD34). PDL stromal cells were differentiated into osteogenic, adipogenic, and chondrogenic lineages. Myotube structures were induced in PDL stromal cells after 5-Aza pretreatment, but not in the absence of 5-Aza pretreatment. CONCLUSIONS: PDL stromal cells isolated from extracted premolars can potentially be a good source of postnatal stem cells for oromaxillofacial regeneration in bone and muscle.
Azacitidine ; Bicuspid ; Bone Marrow ; DNA ; Durapatite ; Fibroblasts ; Hematopoietic Stem Cells ; Humans ; Mesenchymal Stromal Cells ; Molecular Biology ; Muscle Fibers, Skeletal ; Muscles ; Periodontal Ligament ; Regeneration ; Stem Cells ; Stromal Cells

Actin cytoskeleton has been known to control and/or be associated with chondrogenesis. Staurosporine and cytochalasin D modulate actin cytoskeleton and affect chondrogenesis. However, the underlying mechanisms for actin dynamics regulation by these agents are not known well. In the present study, we investigate the effect of staurosporine and cytochalasin D on the actin dynamics as well as possible regulatory mechanisms of actin cytoskeleton modulation. Staurosporine and cytochalasin D have different effects on actin stress fibers in that staurosporine dissolved actin stress fibers while cytochalasin D disrupted them in both stress forming cells and stress fiber-formed cells. Increase in the G-/F-actin ratio either by dissolution or disruption of actin stress fiber is critical for the chondrogenic differentiation. Cytochalasin D reduced the phosphorylation of cofilin, whereas staurosporine showed little effect on cofilin phosphorylation. Either staurosporine or cytochalasin D had little effect on the phosphorylation of myosin light chain. These results suggest that staurosporine and cytochalasin D employ different mechanisms for the regulation of actin dynamics and provide evidence that removal of actin stress fibers is crucial for the chondrogenic differentiation.
Actin Cytoskeleton/*drug effects ; Actins/metabolism ; Animals ; Cell Differentiation/*drug effects ; Cells, Cultured ; Chickens ; Chondrogenesis/*drug effects ; Cytochalasin D/*pharmacology ; Mesoderm/cytology/drug effects ; Myosin Light Chains/metabolism ; Nucleic Acid Synthesis Inhibitors/*pharmacology ; Phosphorylation ; Staurosporine/*pharmacology ; Stress Fibers/drug effects

No abstract available.
Infarction* ; Paralysis*

BACKGROUND AND PURPOSE: Alterations in blood fatty acid (FA) composition are associated with cardiovascular diseases. We investigated whether plasma FA composition was related to stroke severity and functional outcome in acute ischemic stroke patients. METHODS: We prospectively enrolled 156 patients with first-episode cerebral infarction, within 7 days of symptom onset. The proportion of FAs was analyzed using gas chromatography, and the summation of the omega-3 polyunsaturated fatty acids (omega3-PUFA), 18:3 omega3 alpha-linolenic acid, 20:3 omega3 eicosatrienoic acid, 20:5 omega3 eicosapentaenoic acid (EPA), and 22:6 omega3 docosahexaenoic acid (DHA) was reported as Sigmaomega3-PUFAs. Stroke severity was assessed using the National Institutes of Health Stroke Scale (NIHSS) score on admission. Poor functional outcome was defined by modified Rankin scale (mRS) > or =3 at three months after the index stroke. RESULTS: Lower proportions of EPA (beta=-0.751), DHA (beta=-0.610), and Sigmaomega3-PUFAs (beta=-0.462) were independently associated with higher NIHSS score, after adjusting for stroke subtype, hemoglobin, high density lipoprotein, high sensitivity C-reactive protein, fasting glucose, 16:0 palmitic acid, and Sigmasaturated fatty acids. Moreover, a lower proportion of DHA (odds ratio [OR]: 0.20, 95% confidence interval [CI]: 0.04-0.88), and Sigmaomega3-PUFAs (OR: 0.22, 95% CI: 0.05-0.84) showed an independent relationship with poor functional outcome after adjusting for age, sex, smoking status, NIHSS score, stroke subtype, and 16:0 palmitic acid. CONCLUSIONS: Our results demonstrate that omega3-PUFAs correlated with stroke severity on admission and functional outcomes at 3 months. omega3-PUFAs are potential blood biomarkers for prognosis of acute non-cardiogenic ischemic stroke patients.
alpha-Linolenic Acid ; Biomarkers ; C-Reactive Protein ; Cardiovascular Diseases ; Cerebral Infarction ; Chromatography, Gas ; Eicosapentaenoic Acid ; Fasting ; Fatty Acids* ; Fatty Acids, Unsaturated ; Glucose ; Humans ; Lipoproteins ; National Institutes of Health (U.S.) ; Palmitic Acid ; Plasma* ; Prognosis ; Prospective Studies ; Smoke ; Smoking ; Stroke*

BACKGROUND AND PURPOSE: Low-density lipoprotein (LDL) particle size is considered to be one of the more important cardiovascular risk factors, and small LDL particles are known to have atherogenic potential. The aim of this study was to determine whether LDL particle size is associated with stroke severity and functional outcome in patients with atherothrombotic stroke. METHODS: Between January 2009 and May 2011, 248 patients with first-episode cerebral infarction who were admitted to our hospital within 7 days after symptom onset were prospectively enrolled. LDL particle size was measured using the nondenaturing polyacrylamide gradient gel electrophoresis assay. Stroke severity was assessed by applying the National Institutes of Health Stroke Scale (NIHSS) at admission. Functional outcome was investigated at 3 months after the index stroke using the modified Rankin Scale (mRS), and poor functional outcome was defined as an mRS score of > or =3. RESULTS: The LDL particle size in the 248 patients was 25.9+/-0.9 nm (mean+/-SD). LDL particle size was inversely correlated with the degree of cerebral artery stenosis (p=0.010). Multinomial multivariate logistic analysis revealed that after adjustment for age, sex, and variables with p<0.1 in univariate analysis, LDL particle size was independently and inversely associated with stroke severity (NIHSS score > or =5; reference, NIHSS score 0-2; odds ratio=0.38, p=0.028) and poor functional outcome (odds ratio=0.44, p=0.038). CONCLUSIONS: The results of this study demonstrate that small LDL particles are independently correlated with stroke outcomes. LDL particle size is thus a potential biomarker for the prognosis of atherothrombotic stroke.
Atherosclerosis ; Cerebral Arteries ; Cerebral Infarction ; Constriction, Pathologic ; Electrophoresis ; Humans ; Lipoproteins ; National Institutes of Health (U.S.) ; Particle Size* ; Prognosis ; Prospective Studies ; Risk Factors ; Stroke*

BACKGROUND: The aim of this study was to investigate the association between body mass index (BMI) and survival in patients with severe sepsis or septic shock. METHODS: We analyzed the sepsis registry of patients presenting to the emergency department (ED) of a tertiary urban hospital and meeting the criteria for severe sepsis or septic shock from August 2008 to March 2012. We categorized patients into the underweight group (BMI < 18.5 kg/m2), the normal weight group (18.5 < or = BMI < 25 kg/m2) and the obese group (BMI > or = 25 kg/m2). Then, we analyzed the registry to evaluate the relation between obesity and in-hospital mortality. RESULTS: A total of 770 adult patients with severe sepsis and septic shock were analyzed. In-hospital mortality rate of the underweight group (n = 86), the normal weight group (n = 489) and the obese group (n = 195) was 22.1%, 15.3% and 16.4%, respectively. In a multivariate regression analysis, the underweight group had a significant association with in-hospital mortality compared with the normal weight group (odds ratio [OR], 1.12; 95% confidence interval [CI], 0.68-1.87; p = 0.028). The obese group showed no significant difference in mortality (OR, 2.04; 95% CI, 1.08-3.86; p = 0.65). CONCLUSIONS: The underweight patients showed significantly higher mortality than the normal weight patients with severe sepsis and septic shock.
Adult ; Body Mass Index* ; Emergencies ; Hospital Mortality ; Hospitals, Urban ; Humans ; Mortality ; Obesity ; Sepsis* ; Shock, Septic* ; Thinness

Furanocoumarin 8-methoxypsoralen (8-MOP) is the parent compound that naturally occurs in traditional medicinal plants used historically. 8-MOP has been employed as a photochemotherapeutic component of Psoralen + Ultraviolet A (PUVA) therapy for the treatment of vitiligo and psoriasis. Although the role of 8-MOP in PUVA therapy has been studied, little is known about the effects of 8-MOP alone on human gastric cancer cells. In this study, we observed anti-proliferative effect of 8-MOP in several human cancer cell lines. Among these, the human gastric cancer cell line SNU1 is the most sensitive to 8-MOP. 8-MOP treated SNU1 cells showed G1-arrest by upregulating p53 and apoptosis by activating caspase-3 in a dose-dependent manner, which was confirmed by loss-of-function analysis through the knockdown of p53-siRNA and inhibition of apoptosis by Z-VAD-FMK. Moreover, 8-MOPinduced apoptosis is not associated with autophagy or necrosis. The signaling pathway responsible for the effect of 8-MOP on SNU1 cells was confirmed to be related to phosphorylated PI3K, ERK2, and STAT3. In contrast, 8-MOP treatment decreased the expression of the typical metastasis-related proteins MMP-2, MMP-9, and Snail in a p53-independent manner. In accordance with the serendipitous findings, treatment with 8-MOP decreased the wound healing, migration, and invasion ability of cells in a dose-dependent manner. In addition, combination treatment with 8-MOP and gemcitabine was effective at the lowest concentrations. Overall, our findings indicate that oral 8-MOP has the potential to treat early human gastric cancer, with fewer side effects.

Purpose: The purpose of this study was to investigate the concordance rate of PIK3CA mutations between primary and matched distant metastatic sites in patients with breast cancer and to verify whether there are differences in the frequency of PIK3CA hotspot mutations depending on the metastatic sites involved. Materials and Methods: Archived formalin-fixed paraffin-embedded (FFPE) specimens of primary breast and matched distant metastatic tumors were retrospectively obtained for 49 patients. Additionally, 40 archived FFPE specimens were independently collected from different breast cancer metastatic sites, which were limited to three common sites: the liver, brain, and lung. PIK3CA mutations were analyzed using droplet digital PCR, including hotspots involving exons 9 and 20. Results: After analysis of 49 breast tumors with matched metastasis sites, 87.8% showed concordance in PIK3CA mutation status. According to PIK3CA hotspot mutation testing in 89 cases of breast cancer metastatic sites, the proportion of PIK3CA mutations at sites of metastasis involving the liver, brain, and lung was 37.5%, 28.6%, and 42.9%, respectively, which did not result in statistical significance. Conclusion The high concordance of PIK3CA mutation status between primary and matched metastasis sites suggests that metastatic sites, regardless of the metastatic organ, could be considered sample sources for PIK3CA mutation testing for improved therapeutic strategies in patients with metastatic breast cancer.

PURPOSE: The purpose of this study was to investigate the effect of common sources of infection on outcome in patients with severe sepsis and septic shock in the emergency department (ED). METHODS: We conducted a retrospective observational study involving adult patients who were diagnosed with severe sepsis or septic shock in the ED of a tertiary care hospital during the period between August 2008 and March 2012. We categorized patients according to four groups based on source of infection (respiratory infection, intra-abdominal infection [IAI], urinary tract infection [UTI], and other sources [OS] group). The primary outcome was inhospital mortality. Multivariable logistic regression analysis was performed for adjustment of potential confounders, including age, gender, serum lactate concentrations, the Sequential Organ Failure Assessment score, timely antibiotic use, and achievements of early resuscitation targets. RESULTS: A total of 758 patients were included and overall in-hospital mortality was 16.6%. Significant differences in mortality were observed between four groups (27.5% for respiratory infection, 12.1% for IAI, 2.6% for UTI, and 20.0% for other sources, p<0.01). In patients with IAI, adjusted odds ratios (ORs) for mortality were 0.49 (95% confidence interval [CI], 0.27-0.92) compared with the OS group and 0.57 (95% CI, 0.35-0.93) compared with non-IAI. For UTI, adjusted ORs were 0.08 (95% CI, 0.02-0.32) compared with the OS group and 0.09 (95% CI, 0.03-0.35) compared with non-UTI. For respiratory infection, adjusted ORs were 1.33 (95% CI, 0.74-2.39) compared with the OS group and 2.56 (95% CI, 1.60-4.10) compared with non-respiratory infection. CONCLUSION: Results of our study showed that source of infection was independently associated with in-hospital mortality in patients with severe sepsis and septic shock in the ED. In particular, UTI and IAI showed significant association with in-hospital survival. Patients with respiratory infection showed significantly higher mortality, compared with non-respiratory infection patients.
Adult ; Emergency Service, Hospital* ; Hospital Mortality ; Humans ; Intraabdominal Infections ; Lactic Acid ; Logistic Models ; Mortality ; Observational Study ; Odds Ratio ; Resuscitation ; Retrospective Studies ; Sepsis* ; Shock, Septic* ; Tertiary Healthcare ; Urinary Tract Infections

Purpose: To find biomarkers for disease, there have been constant attempts to investigate the genes that differ from those in the disease groups. However, the values that lie outside the overall pattern of a distribution, the outliers, are frequently excluded in traditional analytical methods as they are considered to be ‘some sort of problem.’ Such outliers may have a biologic role in the disease group. Thus, this study explored new biomarker using outlier analysis, and verified the suitability of therapeutic potential of two genes (TM4SF4 and LRRK2). Materials and Methods: Modified Tukey’s fences outlier analysis was carried out to identify new biomarkers using the public gene expression datasets. And we verified the presence of the selected biomarkers in other clinical samples via customized gene expression panels and tissue microarrays. Moreover, a siRNA-based knockdown test was performed to evaluate the impact of the biomarkers on oncogenic phenotypes. Results: TM4SF4 in lung cancer and LRRK2 in breast cancer were chosen as candidates among the genes derived from the analysis. TM4SF4 and LRRK2 were overexpressed in the small number of samples with lung cancer (4.20%) and breast cancer (2.42%), respectively. Knockdown of TM4SF4 and LRRK2 suppressed the growth of lung and breast cancer cell lines. The LRRK2 overexpressing cell lines were more sensitive to LRRK2-IN-1 than the LRRK2 under-expressing cell lines Conclusion Our modified outlier-based analysis method has proved to rescue biomarkers previously missed or unnoticed by traditional analysis showing TM4SF4 and LRRK2 are novel target candidates for lung and breast cancer, respectively.