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Dance is a popular activity throughout the world. There are many styles of dance, each having distinct physical demands and injury patterns. Healthcare providers should have some familiarity with the typical injuries seen in the dancer’s foot and ankle. Along with strong physical exam skills, an understanding of the best imaging options is paramount. High-resolution ultrasound can often be used as reliably as MRI for many of these diagnoses. The present article aims to brief the regional anatomy, address the typical presentation, and review exam findings and management options for the care of dancers’ injuries in the foot and ankle.

Fat grafting, used for soft tissue augmentation during aesthetic or reconstructive plastic surgery, has disadvantages of low efficiency and unpredictable resorption rate. As an alternative, cell-assisted lipotransfer (CAL) is widely used because of its simplicity and low fat resorption rate. However, relevant studies on optimal CAL parameters are still lacking. Here, we aimed to identify the most effective ratio of fat to stromal vascular fraction (SVF) for CAL. We designed two experimental paradigms. The first involved four groups of mice, each group injected with varying ratios of fat and SVF purified from different amounts of fat from a fixed amount of harvested fat. The second experiment involved four groups of mice, each injected with varying amounts of SVF mixed with a fixed amount of fat tissue. The amount of surviving fat in both experiments was compared 8 weeks after fat transplantation. In the first experiment, the group injected with only fat, without consuming any of the harvested fat for SVF purification, showed the greatest mean volume and weight. In the second experiment, groups with 1:1 or more ratio of fat to SVF showed greater volume and weight than the group without SVF. Notably, a ratio of 1:1 did not give significantly different results than higher ratios. Thus, when a limited amount of fat tissue is available, using all of it for grafting is the most effective. However, if an adequate amount is available, using a fat-to-SVF ratio of 1:1 is the most efficient. This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266

Plants regulate a number of primary metabolites, including carbohydrates, organic acids, and amino acids, in response to UV-B radiation. Therefore, it is essential to understand the time-dependent response of rice plants to UV-B stress. This study focused on the response of plants to UV-B at different leaf developmental phases (emerging, growing, and maturing) in an attempt to fully comprehend the metabolic shift. We analyzed the expression levels of genes related to starch/sucrose metabolism in the leaf blades of rice seedlings (Oryza sativa L. “Saechuchenog”) exposed to UV-B irradiation for short (1 day) and long terms (5 days) using quantitative real-time polymerase chain reaction. We also examined the diurnal variations in the contents of primary metabolites using an established GCTOF-MS (gas chromatography time of flight-mass spectrometry) method. The results showed that the levels of primary metabolites were largely dependent upon the diurnal rhythm and leaf developmental phase. The young leaves (sink) produced and accumulated starch rather than sucrose. The short-term (4 h, 1 day) UV-B exposure inhibited sucrose synthesis, which could be the first target of UV-B radiation. Following short- and long-term (5 days) exposure to UV-B radiation, the dynamic response of primary metabolites was evaluated. It was found that the content of carbohydrates decreased throughout the period of exposure to UV-B stress, especially in terms of sucrose concentration. However, the content of the majority of amino acids increased after an early decrease. Our data revealed that the metabolic response, as well as the gene expression, differed with the period (intensity) of exposure to UV-B radiation and with the phase of leaf development. These findings provide new insights for a better understanding of the metabolic response of a variety of plant species exposed to a wide range of UV-B radiation.

With increasing social concerns for healthy food, the studies on the cultivation of crops to increase accumulation of functional metabolites in crops have been investigated. Accumulation of the metabolites in crops is highly affected by various types of stress, such as nutrient deficiency, water deficit (WD), extreme temperature and UV-B radiation as well as their own life cycle. This review summarizes the previous studies on the effects of environmental stresses, especially WD and UV-B radiation, on accumulation of functional metabolites in crops. UV-B radiation and WD during specific period (mainly at maturation stage) activates the adaptation and/or defense system in crops, thereby increasing biosynthesis of the metabolites. Although WD and UV-B radiation tend to decrease in crop yield, the decrease can be compensated by the production of high value crops having high content of functional metabolites.

An estimation of the requirement of minerals based on growth stage and cropping pattern is very important for greenhouse zucchini. This study was performed at farmer’s field which was applied with a fertigation system and a semi-forcing cultivation from Feb. to July in 2014, and nitrogen levels were set up with x0.5, x0.75, x1.0 and x1.5 of the NO₃-N-based soil-testing recommendation for zucchini cultivation. Top dressing of nitrogen (basal : top = 4 : 6) and potassium (basal : top = 3 : 7) was applied with an interval of every two weeks from two and six weeks after transplanting, respectively, and phosphorus was totally supplied with basal dressing. The nitrogen uptake was the order of x1.0, x0.75, x1.5 and x0.5, phosphorus, x1.0, x0.75, x0.5 and x1.5, and potassium, x0.75, x1.0, x1.5 and x0.5. From these results, it was suggested that highest mineral uptake could be reached between x0.75 and x1.0 of the NO₃-N-based soil-testing recommendation. In conclusion, nutrient management based on the growth stage was proven to be better method for favorable growth and yield of zucchini.

This study was conducted to evaluate the effects of reduced nitrogen (N) and potassium (K) fertigation as additional fertilizer on tomato yield and nutrient contents in excessively nutrients-accumulated soil. Shoot and root dry weights (DW), dry matter rate for shoot, root and fruit and number of fruit in both AF50 and AF100 (50 and 100% levels of additional fertilizer) treatments were increased in comparison with those in AF0 (0% level of additional fertilizer) treatment. In case of nutrient uptake by tomato, nitrogen, phosphorous (P) and potassium contents in all tomato parts (leaf, stem, root and fruit) in AF50 and AF100 treatment were lower than those in AF0 treatment. On the contrary, soluble sugar and starch contents in all tomato parts in AF50 and AF100 were higher than those in AF0 treatment. There were differences between AF0 and AF50 or AF100 in tomato growth, yield, nutrient level and contents of soluble sugar and starch. In contrast, the level and initiation point of fertigation did not significantly affect the parameters. Based on our results, the application of properly reduced level of additional fertilizer is possible to maintain the productivity of tomato and alleviate the nutrient accumulation in plastic film house soils.

Leucine-rich repeat kinase 2 (LRRK2) has been identified as a causative gene for Parkinson's disease (PD). LRRK2 contains a kinase and a GTPase domain, both of which provide critical intracellular signal-transduction functions. We showed previously that Rab5b, a small GTPase protein that regulates the motility and fusion of early endosomes, interacts with LRRK2 and co-regulates synaptic vesicle endocytosis. Using recombinant proteins, we show here that LRRK2 phosphorylates Rab5b at its Thr6 residue in in vitro kinase assays with mass spectrophotometry analysis. Phosphorylation of Rab5b by LRRK2 on the threonine residue was confirmed by western analysis using cells stably expressing LRRK2 G2019S. The phosphomimetic T6D mutant exhibited stronger GTPase activity than that of the wild-type Rab5b. In addition, phosphorylation of Rab5b by LRRK2 also exhibited GTPase activity stronger than that of the unphosphorylated Rab5b protein. Two assays testing Rab5's activity, neurite outgrowth analysis and epidermal growth factor receptor degradation assays, showed that Rab5b T6D exhibited phenotypes that were expected to be observed in the inactive Rab5b, including longer neurite length and less degradation of EGFR. These results suggest that LRRK2 kinase activity functions as a Rab5b GTPase activating protein and thus, negatively regulates Rab5b signalling.

New tandem mass spectrometric method coupled with liquid chromatography (LC-MS/MS) has been developed to determine the total concentration of camptothecin derivatives (irinotecan and SN-38) regardless of inter-conversion phenomenon between carboxylate and lactone forms. At first, all sample solutions were acidified for 1h in order to completely convert CPT derivatives into their lactone forms and then CPT derivatives were extracted with organic solution containing diethyl ether and ethyl acetate (2:1, v/v) just after alkalization in the range pH 8.0-8.5 in acid-treated solutions. Analytes were separated on a reverse phase C18 column (150×2.1mm) and eluted isocratically with a mobile phase which consisted of acetonitrile-methanol-buffer (0.1% formic acid, 5mM ammonium formate) (3:4:3, v/v). CPT derivatives were monitored by tandem mass spectrometry in electrospay-positive ionization and multiple reaction mode programmed to the following transitions (m/z): '587.6→167.2' of CPT-11, '393.6→349.3' of SN-38 and '349.4→ 305.2' of CPT. The method was validated to have the proper linearity (r(2)>0.99) over the range of 5-1000ng/ml of CPT-11 and 1-250ng/ml of SN-38 with good accuracy (89.8-114.3%) and precision (less than 10%). In all stability tests, concentration of CPT-11 and SN-38 had been left in the acceptable range of 88.8-110.7% when sample solutions were acidified before determination of CPT derivatives. Newly developed LC-MS/MS method was suitable for the determination of CPT derivatives of both rabbit plasma and tumor tissues in the pharmacokinetic study.

Expected climatic changes likely elicit serious challenges for crop production. Therefore, it is indispensable to investigate the response of crop growth parameters and yield under temperature variability environments. The current experiment on chilli pepper growth was conducted in a field, rain-shelter plastic house, and plastic greenhouse, with accumulated temperatures of 2832 &deg, C, 2967 &deg, C, and 3105 &deg, C in 2017, and 2944 &deg, C, 3091 &deg, C, and 3168 &deg, C in 2018 growing seasons. Based on soil analysis, 132.7 kg ha&minus, 1 (1&times, of livestock manure compost as an optimum and 265.4 kg ha&minus, 1 (2&times, ) as a double amount of organic matter were applied to each simulated temperature condition. The results showed that organic manure application favorably affects the growth attributes and nutrient uptake of chilli pepper with the highest values found in the plastic greenhouse, followed by the rain-shelter house, over the open field cultivation condition. The highest growth of chilli pepper was at the 2&times, rate of organic manure application, whereas the highest yield was found at the 1&times, rate of organic manure application. The application of organic manure at the 1&times, rate in the greenhouse increased root, shoot, and fruit dry weights of chilli pepper by 21.4%, 52.4%, and 79.7%, respectively, compared to the control values. These results indicate that the rational use of organic amendments might be the best solution for chilli pepper production under variable climate conditions.

LRRK2 (leucine-rich repeat kinase 2) has been identified as a gene corresponding to PARK8, an autosomal-dominant gene for familial Parkinson’s disease (PD). LRRK2 pathogenicspecific mutants induce neurotoxicity and shorten neurites. To elucidate the mechanism underlying LRRK2 expression, we constructed the LRRK2-promoter-luciferase reporter and used it for promoter analysis. We found that the glucocorticoid receptor (GR) transactivated LRRK2 in a ligand-dependent manner. Using quantitative RT-PCR and Western analysis, we further showed that treatment with dexamethasone, a synthetic GR ligand, induced LRRK2 expression at both the transcriptional and translational levels, in dopaminergic MN9D cells. Dexamethasone treatment also increased expression of α-synuclein, another PD causative gene, and enhanced transactivation of the α-synuclein promoter-luciferase reporter. In addition, dexamethasone treatment to MN9D cells weakly induced cytotoxicity based on an LDH assay. Because glucocorticoid hormones are secreted in response to stress, our data suggest that stress might be a related factor in the pathogenesis of PD. [BMB Reports 2013; 46(9): 454-459]

Atrial fibrillation (AF) is the most common sustained arrhythmia. AF is caused by inflammation, oxidative stress, and various structural heart diseases. An important key factor in AF promotion is that atrial cardiomyocytes have constitutively active acetylcholine-activated K+ (IKACh) current that is enhanced by the tachycardia. Furthermore, it is known that heat shock proteins (HSPs) are involved in the protection against different forms of cellular stress and recent investigators reported that the HSP inducing compound geranylgeranylacetone (GGA, also known as teprenone) prevents atrial remodeling and attenuates the promotion of AF. Whether GGA and GGA-derivatives prevent the acetylcholine-activated K+ current is unknown. Methods: We examined the effect of HSP inducer GGA and 5 derivatives on IKACh current in mouse atrial cardiomyocytes. In the present study, the IKACh current was recorded using a nystatin-perforated whole cell patch-clamp technique following activation by acetylcholine (10 μM for 2 min). After the measurement of the baseline IKACh current, atrial cardiomyocytes were treated with GGA 10μM or a derivative of GGA (NYK9274, NYK9308, NYK9354, NYK9356 or NYK9376) for 10min and the IKACh current was re-measured. Results: IKACh current in mouse atrial cardiomyocyte treated with acetylcholine (10 μM) was significantly increased, which was markedly attenuated by GGA 10μM treatment (n=5, peak; 54±11.2%, quasi-steady-state; 58±11.4%, p Conclusion: This study provides important evidences that the HSP inducer GGA facilitates its potent antiarrhythmic effect by inhibiting IKAch, which underlies electrical remodeling in mammalian atrial cardiomyocytes. Furthermore, two GGA derivatives, NYK9308 and NYK9356, with improved pharma-chemical properties compared to GGA, reveal similar protective effects compared to GGA, and provide important background information for the use of GGA and GGA-like compounds in patients with AF.

2 Division of Radiation Cancer, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul 139-706, Korea - We have recently re- ported the PRC1 promoter as a promoter candidate to control expression of transcriptionally targeted genes for breast cancer gene therapy. We tested whether the PRC 1 promoter could be also applied for the lung cancer gene therapy. In the transient transfection assay with naked plasmids containing the luciferase fused to the PRC1 promoter, the promoter showed little activity in the normal lung cell line, MRC5. However, in the lung cancer A549 cells, PRC1 showed approximately 30-fold activa- tion which was similar to the survivin promoter, the gene whose promoter has been already re- ported as a candidate for the gene therapy of lung cancer. In viral systems, the PRC1 promoter showed approximately 75% and 66% of transcriptional activity compared to the CMV promoter in the adeno-associated virus (AAV) and the adenovirus (AV) systems, respectively. However, the PRC1 promoter in either AAV or AV showed approximately 20% activity compared to the CMV promoter in the normal lung cells. In addition, human lung tumor xenograft mice showed that the PRC1 promoter activity was as strong as the CMV activity in vivo. Taken together, these results sug- gested that PRC1 might be a potential promoter candidate for transcriptionally targeted lung cancer gene therapy.

The leucine-rich repeat kinase 2 (LRRK2) has been identified as the defective gene at the PARK8 locus causing the autosomal dominant form of Parkinson's disease (PD). Although several LRRK2 mutations were found in familial as well as sporadic PD patients, its physiological functions are not clearly defined. In this study, using yeast two-hybrid screening, we report the identification of Rab5b as an LRRK2-interacting protein. Indeed, our GST pull down and co-immunoprecipitation assays showed that it specifically interacts with LRRK2. In addition, subcellular fractionation and immunocytochemical analyses confirmed that a fraction of both proteins co-localize in synaptic vesicles. Interestingly, we found that alteration of LRRK2 expression by either overexpression or knockdown of endogenous LRRK2 in primary neuronal cells significantly impairs synaptic vesicle endocytosis. Furthermore, this endocytosis defect was rescued by co-expression of functional Rab5b protein, but not by its inactive form. Taken together, we propose that LRRK2, in conjunction with its interaction with Rab5b, plays an important role in synaptic function by modulating the endocytosis of synaptic vesicles.

Leucine-rich repeat kinase 2 (LRRK2) is a gene that, upon mutation, causes autosomal-dominant familial Parkinson’s disease (PD). Yeast two-hybrid screening revealed that Snapin, a SNAP-25 (synaptosomal-associated protein-25) interacting protein, interacts with LRRK2. An in vitro kinase assay exhibited that Snapin is phosphorylated by LRRK2. A glutathione-S-transferase (GST) pull-down assay showed that LRRK2 may interact with Snapin via its Ras-of-complex (ROC) and N-terminal domains, with no significant difference on interaction of Snapin with LRRK2 wild type (WT) or its pathogenic mutants. Further analysis by mutation study revealed that Threonine 117 of Snapin is one of the sites phosphorylated by LRRK2. Furthermore, a Snapin T117D phosphomimetic mutant decreased its interaction with SNAP-25 in the GST pull-down assay. SNAP-25 is a component of the SNARE (Soluble NSF Attachment protein REceptor) complex and is critical for the exocytosis of synaptic vesicles. Incubation of rat brain lysate with recombinant Snapin T117D, but not WT, protein caused decreased interaction of synaptotagmin with the SNARE complex based on a co-immunoprecipitation assay. We further found that LRRK2-dependent phosphorylation of Snapin in the hippocampal neurons resulted in a decrease in the number of readily releasable vesicles and the extent of exocytotic release. Combined, these data suggest that LRRK2 may regulate neurotransmitter release via control of Snapin function by inhibitory phosphorylation. Researchers in Korea have found that a protein linked with Parkinson's disease (PD) may play a role in regulating neuronal signaling. Mutations in the gene encoding the leucine-rich repeat kinase 2 (LRRK2) protein, a kinase whose enhanced enzyme activity is related to PD pathogenesis, are hereditary risk factors for this neurodegenerative disorder. Researchers led by Wongi Seol of Wonkwang University searched for substrates for LRRK2 kinase that might explain this enzyme's involvement in PD. They determined that LRRK2 phosphorylates Snapin, a protein that ensures proper localization of neurotransmitter reserves and LRRK2-mediated phosphorylation inhibits Snapin, thus reducing the size of these reserves and the amount of neurotransmitter release. The extent to which LRRK2-Snapin interaction contributes to Parkinson's remains unclear, but this pathway nevertheless seems to play an important role in proper brain function.

The constitutive androstane receptor (CAR) is a member of the nuclear receptor superfamily and plays an important role in the degradation of xenobiotics in the liver. Using yeast two-hybrid screening, we identified SF3a3, a 60-kDa subunit of the splicing factor 3a complex, as a specific CAR-interacting protein. We further confirmed their interaction by both co-immunoprecipitation and GST pull-down assay. Functional studies showed that overexpression of SF3a3 inhibited the reporter activity driven by a promoter containing CAR binding sequences by up to 50%, whereas reduced expression of SF3a3 activated the same reporter activity by approximately three-fold. The inhibitory function of SF3a3 is independent of the presence of TCPOBOP, a CAR ligand. These data suggest that SF3a3 functions as a co-repressor of CAR transcriptional activity, in addition to its canonical function.

For cancer gene therapy, cancer-specific over- expression of a therapeutic gene is required to reduce side effects derived from expression of the gene in normal cells. To develop such an expression vector, we searched for genes over-expressed and/or specifically expressed in cancer cells using bioinformatics and have selected genes coding for protein regulator of cytokinesis 1 (PRC1) and ribonuclease reductase 2 (RRM2) as candidates. Their cancer-specific expressions were confirmed in both breast cancer cell lines and patient tissues. We compared each promoter's cancer-specific activity in the breast normal and cancer cell lines using the luciferase gene as a reporter and confirmed cancer-specific expression of both PRC1 and RRM2 promoters. To test activities of these promoters in viral vectors, the promoters were also cloned into an adeno-associated viral (AAV) vector containing green fluorescence protein (GFP) as the reporter. The GFP expression levels by these promoters were various depending on cell lines tested and, in MDA-MB-231 cells, GFP activities derived from the PRC1 and RRM2 promoters were as strong as that from the cytomegalovirus (CMV) promoter. Our result showed that a vector containing the PRC1 or RRM2 promoter could be used for breast cancer specific overexpression in gene therapy.