BACKGROUND: Exon deletions of Duchenne muscular dystrophy (DMD) gene account for most of the alterations found in DMD and Becker muscular dystrophy (BMD). This study was to evaluate the usefulness of dual priming oligonucleotide multiplex PCR (DPO PCR) in detection of exon deletions of DMD gene. METHODS: Thirty-seven DMD or BMD patients who had known exon deletions detected by conventional multiplex PCR (conventional PCR) and nine control subjects were enrolled in this study. When a discrepancy was shown between the results of conventional PCR and DPO PCR, the multiplex ligation-dependent probe amplification (MLPA) technique was performed as a confirmation test. RESULTS: The same deletions previously identified by conventional PCR in 32 out of 37 subjects were also detected by DPO PCR. For the five subjects (13.5%) showing discrepant results between the conventional PCR and DPO PCR, MLPA was performed and its results were found to correlate better with those of DPO PCR. The discrepancies were due to false positive or false negative results of the conventional PCR. CONCLUSIONS: DPO PCR shows a high agreement of results with the conventional PCR and is considered an adequate method to be used as a primary genetic test for the diagnosis of DMD. Because of an improved accuracy, especially for determining the boundaries of DMD gene deletions, DPO PCR can be very useful as a supplement to the conventional PCR.
*DNA Mutational Analysis ; DNA Primers ; Dystrophin/*genetics ; Female ; Gene Deletion ; Genetic Screening ; Humans ; Male ; Muscular Dystrophy, Duchenne/*diagnosis/genetics ; Nucleic Acid Amplification Techniques ; Oligonucleotide Probes ; Polymerase Chain Reaction/*methods ; Reagent Kits, Diagnostic ; Reproducibility of Results

Indoleamine 2,3-dioxygenases (IDOs) are tryptophan-catabolizing enzymes with immunomodulatory functions. However, the biological role of IDO2 and its relationship with IDO1 are unknown. To assess the relationship between IDO2 and IDO1, we investigated the effects of co-expression of human (h) IDO2 on hIDO1 activity. Cells co-expressing hIDO1 and hIDO2 showed reduced tryptophan metabolic activity compared with those expressing hIDO1 only. In a proteomic analysis, hIDO1-expressing cells exhibited enhanced expression of proteins related to the cell cycle and amino acid metabolism, and decreased expression of proteins related to cell survival. However, cells co-expressing hIDO1 and hIDO2 showed enhanced expression of negative regulators of cell apoptosis compared with those expressing hIDO1 only. Co-expression of hIDO1 and hIDO2 rescued the cell death induced by tryptophan-depletion through hIDO1 activity. Cells expressing only hIDO2 exhibited no marked differences in proteome profiles or cell growth compared with mock-transfectants. Cellular tryptophan metabolic activity and cell death were restored by co-expressing the hIDO2 mutant substituting the histidine 360 residue for alanine. These results demonstrate that hIDO2 plays a novel role as a negative regulator of hIDO1 by competing for heme-binding with hIDO1, and provide information useful for development of therapeutic strategies to control cancer and immunological disorders that target IDO molecules.
Cell Proliferation ; Cell Survival ; Gene Expression ; HEK293 Cells ; Heme/*metabolism ; Humans ; Indoleamine-Pyrrole 2,3,-Dioxygenase/genetics/*metabolism ; Protein Binding ; Tryptophan/*metabolism ; Up-Regulation

OBJECTIVE: Neuropathic pain causes patients feel indescribable pain. Deep Brain Stimulation (DBS) is one of the treatment methods in neuropathic pain but the action mechanism is still unclear. To study the effect and mechanism of analgesic effects from DBS in neuropathic pain and to enhance the analgesic effect of DBS, we stimulated the ventral posterolateral nucleus (VPL) in rats. METHODS: To observe the effect from VPL stimulation, we established 3 groups : normal group (Normal group), neuropathic pain group (Pain group) and neuropathic pain+DBS group (DBS group). Rats in DBS group subjected to electrical stimulation and the target is VPL. RESULTS: We observed the behavioral changes by DBS in VPL (VPL-DBS) on neuropathic pain rats. In our study, the pain score which is by conventional test method was effectively decreased. In specific, the time of showing withdrawal response from painful stimulation which is not used measuring method in our animal model was also decreased by DBS. CONCLUSION: The VPL is an effective target on pain modulation. Specifically we could demonstrate changes of pain response duration which is not used, and it was also significantly meaningful. We thought that this study would be helpful in understanding the relation between VPL-DBS and neuropathic pain.
Animals ; Deep Brain Stimulation ; Electric Stimulation ; Humans ; Models, Animal ; Neuralgia* ; Rats ; Ventral Thalamic Nuclei

Gastric cancer is one of the most common cancers in the world. The aims of this study were to evaluate the association between polymorphisms in TFF gene family, TFF1, TFF2, and TFF3 and the risk of gastric cancer (GC) and GC subgroups in a Korean population via a case-control study. The eight polymorphisms in TFF gene family were identified by sequencing and genotyped with 377 GC patients and 396 controls by using TaqMan genotyping assay. The rs184432 TT genotype of TFF1 was significantly associated with a reduced risk of GC (odds ratio, [OR) = 0.45; 95% confidence interval, [CI] = 0.25-0.82; P = 0.009), more protective against diffuse-type GC (OR = 0.20; 95% CI = 0.05-0.89; P = 0.035) than GC (OR = 0.34; 95% CI = 0.14-0.82; P = 0.017) in subjects aged < 60 yr, and correlated with lymph node metastasis negative GC and diffuse-type GC (OR = 0.44; 95% CI = 0.23-0.86; P = 0.016 and OR = 0.20; 95% CI = 0.05-0.87; P = 0.031, respectively). In addition, a decreased risk of lymph node metastasis negative GC and diffuse-type GC was observed for rs225359 TT genotype of TFF1 (OR = 0.46, 95% CI = 0.24-0.88; P = 0.020 and OR = 0.21, 95% CI = 0.05-0.88; P = 0.033, respectively). These findings suggest that the rs184432 and rs225359 polymorphisms in TFF1 have protective effects for GC and contribute to the development of GC in Korean individuals.
Adult ; Aged ; Biomarkers, Tumor/*genetics ; Female ; Genetic Markers/genetics ; Genetic Predisposition to Disease/epidemiology/genetics ; Humans ; Incidence ; Male ; Middle Aged ; Peptides/*genetics ; Polymorphism, Single Nucleotide/genetics ; Promoter Regions, Genetic/genetics ; Reproducibility of Results ; Republic of Korea/epidemiology ; Risk Assessment/methods ; Sensitivity and Specificity ; Stomach Neoplasms/*epidemiology/*genetics ; Tumor Suppressor Proteins/*genetics

Background/Aims: Combination therapy with immunomodulators (IMMs) was proposed as a strategy to prevent the development of loss of response (LOR) to anti-tumor necrosis factor (TNF) for patients with inflammatory bowel disease (IBD). However, the effect is unclear in patients already exposed to IMMs. The aim of this study was to evaluate whether combination therapy with IMMs is superior to monotherapy for prevention of LOR to anti-TNF. Methods: This was a retrospective study of patients in Seoul National University Bundang Hospital with IBD between January 2009 and October 2018. LOR was defined as clinical deterioration after maintenance of anti-TNF for at least 6 months. We investigated the difference in incidence of LOR to anti-TNF between the monotherapy and combination groups. We additionally assessed factors affecting LOR development to anti-TNF. Results: A total of 116 patients with IBD were included in this study (monotherapy 61 patients; combination 55 patients). Overall, LOR to anti-TNF occurred in 31 patients during the follow-up period. The combination of an anti-TNF agent and IMM showed no significant difference in the incidence of LOR compared to anti-TNF agent monotherapy (hazard ratio [HR], 1.64; 95% confidence interval [CI], 0.786 to 3.148; p = 0.182). Female sex was significantly associated with the development of LOR to anti-TNF (HR, 3.032; 95% CI, 1.467 to 6.268; p = 0.003). Conclusions Anti-TNF and IMM combination therapy did not prove efficacious in preventing the development of LOR in IBD patients. Female sex was associated with the development of LOR to anti-TNF; further studies are required to confirm these results.

The purpose of this study was to evaluate the effect of the reaction between investment material and zirconia on the strength of zirconia in the application of heat-pressing method. Sixty specimens were cut (24 mm×4 mm×0.5 mm) into plates from Zirtooth ™ Multi O-9814 block (∅98×14T, HASS, Gangwondo, Korea) and sintered at 1450℃. Specimens were divided into 6 subgroups according to the depending on the investement material; (a) UN group (Control), (b) PH group (Prime vest HS), (c) CP group (Calibra-press), (d) BV group (BC-Vest), (e) MH group (Microstar-HS), (f) F1 group (Formula 1). Five investment materials were buried according to the procedure recommended by the manufacturer and left at room temperature for 30 minutes. The investment mold was dried and maintained at an elevated temperature of 850℃ for 50 minutes. Then, Amber Lisi-POZ LT (HASS) was placed in a thermoformed electric furnace (Programat EP3000/G2, Ivoclar Vivadent, Schaan, Liechtenstein) together with the mold, heated to 915℃ at an elevation temperature of 45℃/min, and moored for 15 minutes. The specimens were loaded to fracture in a universal testing machine and the fracture surface was examined by a field-emission scanning electron microscopy (FE-SEM). The surface of the zirconia specimen with the investment material was analyzed by energy dispersive X-ray spectroscopy (EDS). The 3-point flexural strength test showed the highest value (1265.5 MPa) in the UN group and the lowest value (756.1 MPa) in the F1 group. As a result of EDS analysis, the largest amount of Si was detected in the F1 group, and the most interfacial changes occurred as a result of FE-SEM analysis. It was concluded that when the zirconia is buried with the investment material and the heat press molding is performed, the state of the interface is changed due to the investment material at the bonding interface while the strength is lowered.

The purpose of this study was to investigate the effect of glazing treatment of multi-colored zirconia disc and lithium disilicate block on the optical properties. From an Ultra Translucent Multi-Layered (UTML) disc containing 50% cubic crystal and a lithium disilicate-based Amber Mill Direct (AMD) block, the 1.2 mm thick specimens of the upper layer (T), the center (M), and the lower layer (B) were prepared. Three types of paste glaze of InSync Glaze Paste, IPS E.max Ceram Glaze Paste, and Amber Ceram G7 Glaze Paste were applied and fired to a uniform thickness on the surface of the specimen. CIE L * , a * , b * values were measured using a spectrophotometer and contrast and translucency were calculated. The bonding interface with the glaze layer was observed using HR FE-SEM. In the glazed groups, L * value increased, but a * value and b * value decreased. The translucency value measured in the untreated group was statically higher in the AMD groups than in UTML groups (P<0.05).The translucency did not show a significant difference before and after glazing treatment in the UTML group (P>0.05), but significantly decreased compared to the untreated group after glazing treatment in the AMD group (P<0.05). The contrast ratios generally increased after glaze treatment compared to the untreated group. Since the glaze treatment can affect the optical properties of ceramic restorations, the choice of glaze should be considered important for the desired color by the dental technician.

The purpose of this study was to evaluate the effect of the reaction between investment material and zirconia on the strength of zirconia in the application of heat-pressing method. Sixty specimens were cut (24 mm×4 mm×0.5 mm) into plates from Zirtooth ™ Multi O-9814 block (∅98×14T, HASS, Gangwondo, Korea) and sintered at 1450℃. Specimens were divided into 6 subgroups according to the depending on the investement material; (a) UN group (Control), (b) PH group (Prime vest HS), (c) CP group (Calibra-press), (d) BV group (BC-Vest), (e) MH group (Microstar-HS), (f) F1 group (Formula 1). Five investment materials were buried according to the procedure recommended by the manufacturer and left at room temperature for 30 minutes. The investment mold was dried and maintained at an elevated temperature of 850℃ for 50 minutes. Then, Amber Lisi-POZ LT (HASS) was placed in a thermoformed electric furnace (Programat EP3000/G2, Ivoclar Vivadent, Schaan, Liechtenstein) together with the mold, heated to 915℃ at an elevation temperature of 45℃/min, and moored for 15 minutes. The specimens were loaded to fracture in a universal testing machine and the fracture surface was examined by a field-emission scanning electron microscopy (FE-SEM). The surface of the zirconia specimen with the investment material was analyzed by energy dispersive X-ray spectroscopy (EDS). The 3-point flexural strength test showed the highest value (1265.5 MPa) in the UN group and the lowest value (756.1 MPa) in the F1 group. As a result of EDS analysis, the largest amount of Si was detected in the F1 group, and the most interfacial changes occurred as a result of FE-SEM analysis. It was concluded that when the zirconia is buried with the investment material and the heat press molding is performed, the state of the interface is changed due to the investment material at the bonding interface while the strength is lowered.

The purpose of this study was to investigate the effect of glazing treatment of multi-colored zirconia disc and lithium disilicate block on the optical properties. From an Ultra Translucent Multi-Layered (UTML) disc containing 50% cubic crystal and a lithium disilicate-based Amber Mill Direct (AMD) block, the 1.2 mm thick specimens of the upper layer (T), the center (M), and the lower layer (B) were prepared. Three types of paste glaze of InSync Glaze Paste, IPS E.max Ceram Glaze Paste, and Amber Ceram G7 Glaze Paste were applied and fired to a uniform thickness on the surface of the specimen. CIE L * , a * , b * values were measured using a spectrophotometer and contrast and translucency were calculated. The bonding interface with the glaze layer was observed using HR FE-SEM. In the glazed groups, L * value increased, but a * value and b * value decreased. The translucency value measured in the untreated group was statically higher in the AMD groups than in UTML groups (P<0.05).The translucency did not show a significant difference before and after glazing treatment in the UTML group (P>0.05), but significantly decreased compared to the untreated group after glazing treatment in the AMD group (P<0.05). The contrast ratios generally increased after glaze treatment compared to the untreated group. Since the glaze treatment can affect the optical properties of ceramic restorations, the choice of glaze should be considered important for the desired color by the dental technician.

Purpose: Glioblastoma (GBM) is an intractable disease for which various treatments have been attempted, but with little effect.This study aimed to measure the effect of photodynamic therapy (PDT) and sonodynamic therapy (SDT), which are currently being used to treat brain tumors, as well as sono-photodynamic therapy (SPDT), which is the combination of these two. Materials and Methods: Four groups of Sprague-Dawley rats were injected with C6 glioma cells in a cortical region and treated with PDT, SDT, and SPDT. Gd-MRI was monitored weekly and 18F-FDG-PET the day before and 1 week after the treatment. The acoustic power used during sonication was 5.5 W/cm2 using a 0.5-MHz single-element transducer. The 633-nm laser was illuminated at 100 J/cm2 . Oxidative stress and apoptosis markers were evaluated 3 days after treatment using immunohistochemistry (IHC): 4-HNE, 8-OhdG, and Caspase-3. Results: A decrease in tumor volume was observed in MRI imaging 12 days after the treatment in the PDT group (p<0.05), but the SDT group showed a slight increase compared to the 5-Ala group. The high expression rates of reactive oxygen species-related factors, such as 8-OhdG (p<0.001) and Caspase-3 (p<0.001), were observed in the SPDT group compared to other groups in IHC. Conclusion Our findings show that light with sensitizers can inhibit GBM growth, but not ultrasound. Although SPDT did not show the combined effect in MRI, high oxidative stress was observed in IHC. Further studies are needed to investigate the safety parameters to apply ultrasound in GBM.