Sodium bicarbonate cotransporter NBCn1/Slc4a7 affects locomotor activity and hearing in mice.

Despite a widespread expression pattern in the central nervous system, the role of the sodium bicarbonate cotransporter NBCn1/Slc4a7 has not been investigated for locomotor activity, emotion and cognition. Here, we addressed the behavioral consequences of NBCn1 knockout and evaluated hearing and vision that are reportedly impaired in an earlier line of NBCn1 knockout mice and may contribute to behavioral changes. In a circular open field, the knockout mice traveled a shorter distance, especially in the periphery of the chamber, than wildtype littermates. The knockout mice also traveled a shorter total distance in a home cage-like open field. Rearing and grooming behaviors were reduced. The knockout and control mice displayed similar time spent and number of open and closed arms in the elevated plus maze test, indicating negligible change in anxiety. In the Morris water maze test, both groups of mice learned the location of an escape platform within comparable time on the training trials and showed similar platform identification on the probe trial. The knockout mice maintained normal visual responses in the optokinetic drum and produced evoked potentials in response to light stimuli. However, these mice failed to produce auditory evoked potentials. qPCR revealed a robust expression of an alternatively transcribed NBCn1 variant in the knockout mouse retina. These results indicate that NBCn1 deletion leads to reduced locomotor activity in mice by affecting their exploratory behaviors or emotionality. The deletion also causes hearing loss, but its effect on vision varies between different lines of knockout mice.

Validation of the Korean Version of the Body Image Disturbance Questionnaire-Scoliosis.

STUDY DESIGN: Cross-cultural adaptation and validation of the Body Image Disturbance Questionnaire-Scoliosis OBJECTIVE.: The purpose of this study was to evaluate the reliability and validity of an adapted Korean version of the Body Image Disturbance Questionnaire-Scoliosis version (BIDQ-S). SUMMARY OF BACKGROUND DATA: A modified version of the BIDQ instrument has been validated in adolescent idiopathic scoliosis (AIS) to assess the perception of spinal appearance and psychological disturbance. However, there is no culturally adapted, reliable, and validated BIDQ-S for the Korean population. METHODS: Translation/retranslation of the English version of the BIDQ-S was conducted, and all steps of the cross-cultural adaptation process were performed. The Korean version of the BIDQ-S (K-BIDQ-S) and the previously validated appearance domain of the Korean version of the Scoliosis Research Society-22 Outcomes questionnaire (K-SRS-22) and Spinal Appearance Questionnaire (K-SAQ) were mailed to 152 patients with AIS. Reliability assessments were conducted using κ statistics to assess item agreements, and intraclass correlation coefficients (ICCs) and Cronbach α values were calculated. Convergent validity was evaluated by comparing the scores of the K-BIDQ-S, K-SAQ, and K-SRS-22 appearance domain and discriminant validity by analyzing relationships between K-BIDQ-S score and patient characteristics. RESULTS: All items of the K-BIDQ-S had κ values of agreement >0.6. The K-BIDQ-S showed excellent test/retest reliability with an ICC of 0.912. Internal consistency of the K-BIDQ-S was found to be very good (α = 0.880). Convergent validity testing demonstrated good correlations between the K-BIDQ-S and K-SAQ (r = 0.617), and between the K-BIDQ-S and K-SRS-22 (r = -651). The correlation between the K-BIDQ-S and major curve magnitude was significant (r = 0.688). Discriminant validity was confirmed by significant differences in K-BIDQ-S scores among patients requiring observation, bracing, or surgery. CONCLUSION: The K-BIDQ-S showed satisfactory reliability and validity, and thus, is considered suitable for the evaluation of spinal deformity appearance in Korean-speaking patients with AIS. LEVEL OF EVIDENCE: 3.

Activation of tumor-promoting pathways implicated in hepatocellular adenoma/carcinoma, a long-term complication of glycogen storage disease type Ia.

Hepatocellular adenoma/carcinoma (HCA/HCC) is a long-term complication of the metabolic disorder glycogen storage disease type Ia (GSD-Ia) deficient in glucose-6-phosphatase-α (G6PC or G6Pase-α). We have shown previously that hepatic G6Pase-α deficiency leads to autophagy impairment, mitochondrial dysfunction, enhanced glycolysis, and augmented hexose monophosphate shunt, all of which can contribute to hepatocarcinogenesis. However, the mechanism underlying HCA/HCC development in GSD-Ia remains unclear. We now show that G6Pase-α deficiency-mediated hepatic autophagy impairment leads to sustained accumulation of an autophagy-specific substrate p62 which can activate tumor-promoting pathways including nuclear factor erythroid 2-related factor 2 (Nrf2) and mammalian target of rapamycin complex 1 (mTORC1). Consistently, the HCA/HCC lesions developed in the G6Pase-α-deficient livers display marked accumulation of p62 aggregates and phosphorylated p62 along with activation of Nrf2 and mTORC1 signaling. Furthermore, the HCA/HCC lesions exhibit activation of additional oncogenic pathways, ß-catenin and Yes-associated protein (YAP) which is implicated in autophagy impairment. Intriguingly, hepatic levels of glucose-6-phosphate and glycogen which are accumulated in the G6Pase-α-deficient livers were significantly lower in HCC than those in HCA. Conversely, compared to HCA, the HCC lesion display increased expression of many oncogenes and the M2 isoform of pyruvate kinase (PKM2), a glycolytic enzyme critical for aerobic glycolysis and tumorigenesis. Collectively, our data show that hepatic G6Pase-α-deficiency leads to persistent autophagy impairment and activation of multiple tumor-promoting pathways that contribute to HCA/HCC development in GSD-Ia.

Leptin in adolescent idiopathic scoliosis - A meta-analysis.

To compare the serum levels of leptin and soluble leptin receptor (sOB-R) with adolescent idiopathic scoliosis (AIS) girls and controls through meta-analysis. The MEDLINE via PubMed, Cochrane, Scopus, and EMBASE database, from the earliest available date of indexing between January 2010 and January 2019, were searched for comparative studies evaluating serum levels of leptin and sOB-R in AIS girls. Two authors performed the data extraction independently. Any discrepancies were resolved by a consensus. Six comparative studies were identified. There was no statistically significant difference in terms of leptin between AIS girls and control [p = 0.19, WMD = -2.06 (-5.14, 1.03) ng/mL]. However, the sOB-R level was significantly higher [p < 0.00001, WMD = 2.85 (1.81, 3.88) ng/mL] and the free leptin index was significantly lower [p = 0.0006, WMD = -0.12 (-0.19, -0.05)] in AIS girls than those of healthy control girls. The body mass index was significantly lower in AIS girls [p = 0.03, WMD = -1.53 (-2.95, -0.12) kg/m2]. The current meta-analysis showed that the level of sOB-R is higher in AIS patients than controls, while the concentration of leptin remains unchanged in AIS patients. Further well-designed studies would be necessary to substantiate our results.

Enrichment of vascular endothelial growth factor secreting mesenchymal stromal cells enhances therapeutic angiogenesis in a mouse model of hind limb ischemia.

Critical limb ischemia, a severe manifestation of peripheral artery disease, is emerging as a major concern in aging societies worldwide. Notably, cell-based gene therapy to induce angiogenesis in ischemic tissue has been investigated as treatment. Despite many studies demonstrating the efficacy of this approach, better therapies are required to prevent serious sequelae such as claudication, amputation and other cardiovascular events. We have now established a simplified method to enhance the effects of therapeutic transgenes by selecting for and transplanting only transduced cells. Herein, mesenchymal stromal cells were transfected to co-express vascular endothelial growth factor as angiogenic factor and enhanced green fluorescent protein as marker. Transfected cells were then collected using flow cytometry based on green fluorescence and transplanted into ischemic hind limbs in mice. Compared with unsorted or untransfected cells, purified cells significantly improved blood perfusion within 21days, suggesting that transplanting only cells that overexpress vascular endothelial growth factor enhances therapeutic angiogenesis. Importantly, this approach may prove to be useful in cell-based gene therapy against a wide spectrum of diseases, simply by replacing the gene to be delivered or the cell to be transplanted.

Cardiac Usage of Reducible Poly(oligo-D-arginine) As a Gene Carrier for Vascular Endothelial Growth Factor Expression.

Developments of non-viral carriers have headed toward reducing cytotoxicity, which results from the use of conventional gene carriers, and enhancing gene delivery efficiency. Cys-(d-R9)-Cys repeated reducible poly(oligo-D-arginine) (rPOA) is one of the most efficient non-viral carriers for gene therapy; however, while its efficiency has been verified in the lung and brain, it is necessary to confirm its activity in each organ or tissue since there are differences of gene carrier susceptibility to among tissue types. We therefore tested the compatibility of rPOA in cardiac tissue by in vitro or in vivo experiments and confirmed its high transfection efficiency and low cytotoxicity. Moreover, substantial regenerative effects were observed following transfection with rPOA/pVEGF expression vector complexes (79% decreased infarct size) compared to polyethyleneimine (PEI) (34% decreased infarct size) in a rat myocardial infarction (MI) model. These findings suggest that rPOA efficiently enables DNA transfection in cardiac tissue and can be used as a useful non-viral therapeutic gene carrier for gene therapy in ischemic heart disease.

Urethroplasty by use of turnover flaps (modified mathieu procedure) for distal hypospadias repair in adolescents: comparison with the tubularized incised plate procedure.

PURPOSE: The purpose of this study was to examine whether urethroplasty with a turnover flap, as an alternative method of distal hypospadias repair in adolescents, improves the outcome of surgery. MATERIALS AND METHODS: Between January 2004 and December 2013, a total of 38 adolescents (aged 11-17 years) underwent distal hypospadias repair with either the tubularized incised plate (TIP) procedure (n=25) or the turnover flap procedure (n=13). The turnover flap procedure was performed with a proximal, ventral penile flap that was turned over to cover the urethral plate. Patient demographics, perioperative outcomes, complications, and postoperative uroflowmetry in each surgical group were analyzed retrospectively. RESULTS: The patient demographics were similar in the two groups. There were no significant differences in perioperative outcomes between the groups, including mean operative time, duration of hospital stay, and urethral catheterization. The number of patients with at least one complication, including wound dehiscence, urethrocutaneous fistula, meatal stenosis, and urethral stricture, was lower in the turnover flap group (1/13, 7.7%) than in the TIP group (11/25, 44%, p=0.030). The incidence of meatal stenosis was lower in the turnover flap group (0/12, 0%) than in the TIP group (6/25, 24%). In postoperative uroflowmetry, the plateau-shaped curve rate was lower in the turnover flap group (1/12, 8.3%) than in the TIP group (5/19, 26.3%); the peak flow was higher (p=0.030). CONCLUSIONS: The turnover flap procedure is clinically useful for repairing adolescent distal hypospadias because it offers lower complication rates and better functional outcomes than TIP.

Identification of the cis-element and bZIP DNA binding motifs for the autogenous negative control of mouse NOSTRIN.

mNOSTRIN is the mouse ortholog of hNOSTRIN. Unlike hNOSTRIN, which is alternatively spliced to produce two isoforms (α and ß), only a single isoform of mNOSTRIN has been detected in either the nucleus or cytoplasm/membrane. Because mNOSTRIN represses its own transcription through direct binding onto its own promoter, this protein is constantly expressed in a temporally regulated pattern during differentiation of F9 embryonic carcinoma cells. In this study, we identified the specific cis-element in the mNOSTRIN regulatory region that is responsible for negative autogenous control. This element exhibits inverted dyad symmetry. Furthermore, we identified a putative bZIP motif in the middle region of mNOSTRIN, which is responsible for DNA binding, and showed that disruption of the leucine zippers abolished the DNA-binding activity of mNOSTRIN. Here, we report that a single form of mNOSTRIN functions in both the nucleus and cytoplasm/membrane. In the nucleus, mNOSTRIN acts as a transcriptional repressor by binding to the cis-element through its bZIP motif.

Role of prostate-specific antigen change ratio at initial biopsy as a novel decision-making marker for repeat prostate biopsy.

PURPOSE: Prostate biopsy is used to confirm the prostate cancer. Although first biopsy result was benign, repeat biopsy is recommended for the patient who has higher risk of prostate cancer. In this study, we investigated the PSA change ratio (post-biopsy PSA to baseline PSA) whether it could be predictive factor of prostate cancer and helpful when decided to perform repeat biopsy. MATERIALS AND METHODS: 151 patients, first diagnosed as benign, but underwent repeat biopsy due to clinical suspicion of prostate cancer were included. Post-biopsy PSA was checked 60 minutes later after biopsy. PSA change ratio was defined as post-biopsy PSA to baseline PSA. According to results of repeat biopsy, patients were divided into benign group (group A) and cancer groups (group B). Between two group baseline PSA, PSA density, post-biopsy PSA and PSA change ratio were compared, and most effective cut-off value was analyzed using receiver operating characteristic (ROC). RESULTS: 129 men were benign, 22 men were prostate cancer according to results of repeat biopsy. Between two groups, post-biopsy PSA and PSA change ratio were statically significant differences. (p<0.001, <0.001) The effective cut-off value was 3.0, 3.5 and 4.0 according to ROC. At ROC curve, PSA change ratio was statistically significant for diagnosis of prostate cancer. (AUC 0.800, p<0.001). CONCLUSIONS: PSA change ratio is thought be a predictive factor for prostate cancer. If the PSA change ratio was less than 3.0-4.0, repeat biopsy should be considered to confirm the diagnosis.

Feed-back regulation of disabled-2 (Dab2) p96 isoform for GATA-4 during differentiation of F9 cells.

F9 embryonic carcinoma (EC) cells undergo extra-embryonic endodermal (ExE) differentiation in response to retinoic acid (RA) treatment, which induces the expression of two isoforms (p96 and p67) of the adaptor protein, Disabled-2 (Dab2). In the current study, constitutive and ectopic expression of the p96 isoform induced ExE differentiation in F9 EC cells in the absence of RA treatment via the activation of GATA-4 by p96. During the RA-induced differentiation process, Dab2 expression is induced by the GATA factors in a coherent feed-forward loop; on the other hand, we showed that p96 regulates GATA-4 in a positive feed-back manner in this study. Our results indicate that p96 Dab2 plays a key role in the ExE differentiation process.

Mouse disabled 2 interacting protein 2 functions as a transcriptional repressor through direct binding onto its own promoter.

The mDaIP2 protein is a mouse orthologue of human Nostrin (a regulator of eNos). The absence of eNos activity in RA-treated F9 cell implies that the protein plays somehow different role from Nostrin. In this experiment, this protein has been shown to repress the expression of its own gene, via a feedback mechanism which involves binding to the promoter region. Data from cotransfection, DNAP, mDaIP2-silenced F9 cell, and EMSA analyses clearly support the repression activity and direct binding of the protein to the promoter region. The protein contains N-terminal FCH domain and C-terminal SH3 domain. The SH3 domain is known to interact with the proline-rich domain of mDab2, while even no function has been reported about the FCH domain. Here, we report a novel function of mDaIP2 as a transcriptional repressor and suggest the possible association of the FCH domain with transcriptional regulation.