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RNA degradation is critical for control of gene expression, and endonucleolytic cleavage– dependent RNA degradation is conserved among eukaryotes. Some cleavage sites are secondarily capped in the cytoplasm and identified using the CAGE method. Although uncapped cleavage sites are widespread in eukaryotes, comparatively little information has been obtained about these sites using CAGE-based degradome analysis. Previously, we developed the truncated RNA-end sequencing (TREseq) method in plant species and used it to acquire comprehensive information about uncapped cleavage sites; we observed G-rich sequences near cleavage sites. However, it remains unclear whether this finding is general to other eukaryotes. In this study, we conducted TREseq analyses in fruit flies (Drosophila melanogaster) and budding yeast (Saccharomyces cerevisiae). The results revealed specific sequence features related to RNA cleavage in D. melanogaster and S. cerevisiae that were similar to sequence patterns in Arabidopsis thaliana. Although previous studies suggest that ribosome movements are important for determining cleavage position, feature selection using a random forest classifier showed that sequences around cleavage sites were major determinant for cleaved or uncleaved sites. Together, our results suggest that sequence features around cleavage sites are critical for determining cleavage position, and that sequence-specific endonucleolytic cleavage–dependent RNA degradation is highly conserved across eukaryotes.

Peroneal tendon dislocation (PTD) is sometimes diagnosed as a sports-related injury, of which many cases lead to recurrent PTD (RPTD). Superior retinaculum repair is the major operative treatment of RPTD. The technique described herein comprises 8 steps: (1) Standard tendoscopic examination, (2) debridement of the pseudo-pouch base, (3) first anchor insertion, (4) suture relay, (5) second anchor insertion and suture relay, (6) third anchor insertion and suture relay, (7) suture tightening, and (8) suture bridge. Although this tendoscopic peroneal retinaculum repair technique is complicated, expensive, and requires a longer operation time, it incorporates a double-row suture bridge. Therefore, it has a wider contact surface between the superior retinaculum repair and fibula bone and tighter fixation than does a single-row technique. Moreover, our technique is knotless and thereby avoids knot-related complications. Tendoscopy has additional advantages in terms of less postoperative pain, fewer complications, and better cosmesis. In conclusion, this knotless tendoscopic peroneal retinaculum repair technique for RPTD is a patient-friendly surgery compared with previous procedures.

Plant-made oral vaccines can be a cost-effective method to control infectious diseases of humans and farm animals. Pig edema is a bacterial disease caused by enterohemorrhagic Escherichia coli producing the toxin Shiga toxin 2e (Stx2e). In our previous report, we chose the non-toxic B subunit of Stx2e (Stx2eB) as a vaccine antigen, and Stx2eB was expressed in lettuce (Lactuca sativa L., cv. Green wave). We found that a double repeated Stx2eB (2×Stx2eB) accumulates to higher levels than a single Stx2eB. In this study, we analyzed progeny plants introduced with 2×Stx2eB in which the gene was expressed under the control of conventional cauliflower mosaic virus 35S RNA (CaMV 35S) promoter, and found that the lettuce underwent transgene silencing and bore few seeds. We resolved these problems by using a transgene cassette which harbored a transcriptional promoter derived from the lettuce ubiquitin gene and a longer version of HSPT. The lettuce harboring this expression construct will be valuable in establishing the seed lot system on the basis that thousands of seeds can be obtained from one plant body and the resulting progeny plants accumulate 2×Stx2eB at high levels without the transgene silencing.

The purpose of this study was to evaluate whether tendoscopic peroneal retinaculum repair for patients with recurrent peroneal tendon dislocation (RPTD) is more useful than an open procedure. Twenty-five patients with RPTD were retrospectively reviewed. Twelve patients (13 ankles) with RPTD underwent the open procedure (Group A) between 2008 and 2014, and 13 patients (14 ankles) underwent the tendoscopic procedure (Group B) between 2014 and 2017. Evaluation parameters included clinical results [the Japanese Society for Surgery of the Foot (JSSF) ankle-hind foot scale], operation time, complications, return to sports, and recurrence. Postoperative JSSF ankle/hindfoot scale scores were significantly better than the pre-surgical scores in both groups. The mean operation time was significantly longer in Group B than in Group A (75.7 ± 20.5 vs 38.4 ± 10.5 min). There was one recurrence in Group A, but none in Group B. Group A had no complications, and Group B had one wound infection. Group B, excluding the case of infection, could return to sports earlier than Group A, excluding the recurrent case (13.4 ± 1.5 vs 12.2 ± 0.6 weeks). This tendoscopic procedure needs longer operation time and is more technically demanding, but it is a useful procedure, because it is less invasive and can accelerate return to sports. III.

VASCULAR-RELATED NAC-DOMAIN7 (VND7) is the master transcription factor for vessel element differentiation in Arabidopsis thaliana. To identify the cis-acting sequence(s) bound by VND7, we employed fluorescence correlation spectroscopy (FCS) to find VND7-DNA interactions quantitatively. This identified an 18-bp sequence from the promoter of XYLEM CYSTEINE PEPTIDASE1 (XCP1), a direct target of VND7. A quantitative assay for binding affinity between VND7 and the 18-bp sequence revealed the core nucleotides contributing to specific binding between VND7 and the 18-bp sequence. Moreover, by combining the systematic evolution of ligands by exponential enrichment (SELEX) technique with known consensus sequences, we defined a motif termed the Ideal Core Structure for binding by VND7 (ICSV). We also used FCS to search for VND7 binding sequences in the promoter regions of other direct targets. Taking these data together, we proposed that VND7 preferentially binds to the ICSV sequence. Additionally, we found that substitutions among the core nucleotides affected transcriptional regulation by VND7 in vivo, indicating that the core nucleotides contribute to vessel-element-specific gene expression. Furthermore, our results demonstrate that FCS is a powerful tool for unveiling the DNA-binding properties of transcription factors.

High expression of a transgene is often necessary to produce useful substances in plants. The efficiency of mRNA translation is an important determinant of the level of transgene expression. In dicotyledonous plants, the 5′UTR of certain mRNAs act as translational enhancers, dramatically improving transgene expression levels. On the other hand, translation enhancers derived from dicotyledonous plants are not so much effective in monocotyledonous plants, which are important as industrial crops and as hosts for production of useful substances. In this study, we evaluated the polysome association on a large scale with high resolution for each 5′UTR variant from multiple transcription start site in normal and heat-stressed Oryza sativa suspension cultures. Translational enhancer candidates were selected from the resultant large-scale data set, and their enhancer activities were evaluated by transient expression assay. In this manner, we obtained several translational enhancers with significantly higher activities than previously reported enhancers. Their activities were confirmed in a different monocotyledonous plant, Secale cereale, and using a different reporter gene. In addition, enhancer activities of tested 5′UTRs were different between monocotyledonous and dicotyledonous plants, suggesting that the enhancer activities were not compatible between them. Overall, we demonstrate these useful 5′UTRs as enhancer sequence for transgene expression in monocotyledonous plants.

Background RNA degradation is important for the regulation of gene expression. Despite the identification of proteins and sequences related to deadenylation-dependent RNA degradation in plants, endonucleolytic cleavage-dependent RNA degradation has not been studied in detail. Here, we developed truncated RNA end sequencing in Arabidopsis thaliana to identify cleavage sites and evaluate the efficiency of cleavage at each site. Although several features are related to RNA cleavage efficiency, the effect of each feature on cleavage efficiency has not been evaluated by considering multiple putative determinants in A. thaliana. Results Cleavage site information was acquired from a previous study, and cleavage efficiency at the site level (CSsite value), which indicates the number of reads at each cleavage site normalized to RNA abundance, was calculated. To identify features related to cleavage efficiency at the site level, multiple putative determinants (features) were used to perform feature selection using the Least Absolute Shrinkage and Selection Operator (LASSO) regression model. The results indicated that whole RNA features were important for the CSsite value, in addition to features around cleavage sites. Whole RNA features related to the translation process and nucleotide frequency around cleavage sites were major determinants of cleavage efficiency. The results were verified in a model constructed using only sequence features, which showed that the prediction accuracy was similar to that determined using all features including the translation process, suggesting that cleavage efficiency can be predicted using only sequence information. The LASSO regression model was validated in exogenous genes, which showed that the model constructed using only sequence information can predict cleavage efficiency in both endogenous and exogenous genes. Conclusions Feature selection using the LASSO regression model in A. thaliana identified 155 features. Correlation coefficients revealed that whole RNA features are important for determining cleavage efficiency in addition to features around the cleavage sites. The LASSO regression model can predict cleavage efficiency in endogenous and exogenous genes using only sequence information. The model revealed the significance of the effect of multiple determinants on cleavage efficiency, suggesting that sequence features are important for RNA degradation mechanisms in A. thaliana.

Acute patellar tendon rupture is a serious injury, resulting in the disruption of the knee extensor mechanism. Many authors recommend augmented repairs of patellar tendon ruptures to allow early active rehabilitation. An internal brace technique, which is a ligament augmentation using high-strength suture tape and knotless anchors, has been used as augmentation for the primary tendon or ligament injury. A case of acute patellar tendon rupture in a Judo player, who was successfully treated with primary repair and augmentation using an internal brace technique, is presented. In this case, the patient regained full function of the knee and returned to full sports activities postoperatively. An internal brace technique provides biomechanical stability of the repaired tendon without donor site morbidity and could be an effective procedure for the treatment of acute patellar tendon rupture.

RNA degradation is an important process for controlling gene expression and is mediated by decapping / deadenylation-dependent or endonucleolytic cleavage-dependent RNA degradation mechanisms. High-throughput sequencing of RNA degradation intermediates was initially developed in Arabidopsis thaliana and similar RNA degradome sequencing methods were conducted in other eukaryotes. However, interpreting results obtained by these sequencing methods is fragmented, and an overview is needed. Here we review the findings and limitations of these sequencing methods and discuss the missing experiments needed to understand RNA degradation intermediates accurately. This review provides direction for future research on RNA degradation and is a reference for RNA degradome studies in other species.

Lateral ankle sprains are very common injuries that sometimes lead to chronic lateral ankle instability. The modified Broström operation is the gold standard procedure for treatment of chronic lateral ankle instability. Currently, this operation is performed arthroscopically. Broström repair depends on the quality of the remnant ligament. In cases with an insufficient remnant ligament, Gould augmentation or reconstruction using the gracilis tendon is generally performed. Recently, tape augmentation (internal brace) also has been used to support an insufficient ligament. This article introduces arthroscopic tape augmentation with arthroscopic modified Broström operation. This technique consists of creation of a talar anchor hole and fibular anchor hole, reattachment of the remnant ligament to the fibula with tape, and tape fixation to the talus. This technique uses only one knotless anchor screwed to the fibula for both the modified Broström operation and fixation of the tape. This technique is relatively simple and produces similar results as an open procedure., Technique Video Video 1 This video demonstrates each step of the arthroscopic internal brace augmentation with arthroscopic modified Broström operation. The procedure is performed with the patient in the supine position. The patient's leg is placed on the leg holder, and the hip and knee joints are flexed to about 45°. For creation of the talar anchor hole, the viewing portal and working portal are the anteromedial and accessory anterolateral portals, respectively. For the other procedures, the viewing and working portals are the anterolateral and accessory anterolateral portals, respectively. Surgical steps consist of creation of the talar anchor hole (1:19-1:41), creation of the fibular anchor hole (1:41-1:54), suture lasso using an 18-G needle and 2-0 PROLENE (1:54-2:09), suture relay from the 2-0 PROLENE to a tape (2:09-2:17), grasping of the ATFL and CFL complex by the tape (2:17-2:35), suture lasso insertion for the artificial ligament (2:36-2:45), insertion of the anchor into the fibula with an artificial ligament and tape (2:46-3:15), suture relay from the 2-0 PROLENE to the artificial ligament (3:20-3:28), and insertion of the knotless anchor into the talus (3:35-3:55). (ATFL, anterior talofibular ligament; CFL, calcaneofibular ligament.)

Multiple mechanisms are involved in gene expression, with mRNA degradation being critical for the control of mRNA accumulation. In plants, although some trans-acting factors and motif sequences have been identified in deadenylation-dependent mRNA degradation, endonucleolytic cleavage-dependent mRNA degradation has not been studied in detail. Previously, we developed truncated RNA-end sequencing (TREseq) in Arabidopsis thaliana and detected G-rich sequence motifs around 5′ degradation intermediates. However, it remained to be elucidated whether degradation efficiencies of 5′ degradation intermediates in A. thaliana vary among growth conditions and developmental stages. To address this issue, we conducted TREseq of cultured cells under heat stress and at three developmental stages (seedlings, expanding leaves and expanded leaves) and compared 5′ degradation intermediates data among the samples. Although some 5′ degradation intermediates had almost identical degradation efficiencies, others differed among conditions. We focused on the genes and sites whose degradation efficiencies differed. Changes in degradation efficiencies at the gene and site levels revealed an effect on mRNA accumulation in all comparisons. These changes in degradation efficiencies involved multiple determinants, including mRNA length and translation efficiency. These results suggest that several determinants govern the efficiency of mRNA degradation in plants, helping the organism to adapt to varying conditions by controlling mRNA accumulation.

Objectives Locomotive syndrome (LS) is the leading cause of persons needing long-term care in old age and is characterized by locomotive organ impairment including musculoskeletal pain. The aim was to examine the association between musculoskeletal pain and LS in young and middle-aged persons. Methods A total of 836 participants (male 667, female 169; mean age 44.4 years) were examined in this cross-sectional study. The LS was evaluated by three screening tools: the two-step test, the stand-up test, and the 25-question Geriatric Locomotive Function Scale. Musculoskeletal pain, exercise habits, physical function (walkability and muscle strength), and physical activity were also assessed. Results The LS was found in 22.8% of participants. The number with musculoskeletal pain was significantly higher in those with the LS. A significant correlation was found between the degree of musculoskeletal pain and exercise habits. Less regular exercise was significantly associated with higher LS prevalence. Physical activity and function were greater in participants with more regular exercise. Conclusion Musculoskeletal pain was significantly related to LS even in young and middle-aged persons. The present results suggest that control of musculoskeletal pain and improvement of exercise habits in young and middle-aged persons might help prevent the LS.

Plant high-affinity K(+) (HAK) transporters are divided into four major clusters. Cluster I transporters, in particular, are thought to have high-affinity for K(+). Of the 27 HAK genes in rice, eight HAK transporters belong to cluster I. In this study, we investigated the temporal expression patterns during K(+) deficiency and K(+) transport activity of these eight HAK transporters. The expression of seven HAK genes except OsHAK20 was detected. Expression of OsHAK1, OsHAK5 and OsHAK21 was induced in response to K(+) deficiency; however, that of other genes was not. Six of the eight HAK transporters—OsHAK1, OsHAK5, OsHAK19, OsHAK20, OsHAK21, and OsHAK27—complemented the K(+)-transporter-deficient yeast or bacterial strain. Further, the yeast cells expressing OsHAK1 were more sensitive to Na(+) than those expressing OsHAK5. Mutant analysis showed that the high-affinity K(+) uptake activity was almost undetectable in oshak1 mutants in a low-K(+) medium (0.02 mM). In addition, the high-affinity K(+) uptake activity of wild-type plants was inhibited by mild salt stress (20 mM NaCl); however, Na(+) permeability of OsHAK1 was not detected in Escherichia coli cells. The high-affinity K(+) uptake activity by leaf blades was detected in wild-type plants, while it was not detected in oshak1 mutants. Our results suggest that OsHAK1 and OsHAK5 are the two important components of cluster I corresponding to low-K(+) conditions, and that the transport activity of OsHAK1, unlike that of OsHAK5, is sensitive to Na(+). Further, OsHAK1 is suggested to involve in foliar K(+) uptake.

Plant high-affinity K+ (HAK) transporters are divided into four major clusters. Cluster I transporters, in particular, are thought to have high-affinity for K+. Of the 27 HAK genes in rice, eight HAK transporters belong to cluster I. In this study, we investigated the temporal expression patterns during K+ deficiency and K+ transport activity of these eight HAK transporters. The expression of seven HAK genes except OsHAK20 was detected. Expression of OsHAK1, OsHAK5 and OsHAK21 was induced in response to K+ deficiency; however, that of other genes was not. Six of the eight HAK transporters-OsHAK1, OsHAK5, OsHAK19, OsHAK20, OsHAK21, and OsHAK27-complemented the K+-transporter-deficient yeast or bacterial strain. Further, the yeast cells expressing OsHAK1 were more sensitive to Na+ than those expressing OsHAK5. Mutant analysis showed that the high-affinity K+ uptake activity was almost undetectable in oshak1 mutants in a low-K+ medium (0.02 mM). In addition, the high-affinity K+ uptake activity of wild-type plants was inhibited by mild salt stress (20 mM NaCl); however, Na+ permeability of OsHAK1 was not detected in Escherichia coli cells. The high-affinity K+ uptake activity by leaf blades was detected in wild-type plants, while it was not detected in oshak1 mutants. Our results suggest that OsHAK1 and OsHAK5 are the two important components of cluster I corresponding to low-K+ conditions, and that the transport activity of OsHAK1, unlike that of OsHAK5, is sensitive to Na+. Further, OsHAK1 is suggested to involve in foliar K+ uptake.

Background Despite recommendations on how to prevent baseball injuries in youths by the Japanese Society of Clinical Sports Medicine, shoulder and elbow pain still frequently occurs in young baseball players. We conducted a questionnaire survey among baseball players at elementary schools across the country to understand the practice conditions of players, examining the risk factors of shoulder and elbow pain in baseball players. Methods The questionnaire survey was conducted among elementary school baseball players as members of the Baseball Federation of Japan in September 2015. Results A total of 8354 players belonging to 412 teams (average age: 8.9) responded to the survey. Among 7894 players who did not have any shoulder and/or elbow pain in September 2014, elbow pain was experienced in 12.3% of them, shoulder pain in 8.0% and shoulder and/or elbow pain in 17.4% during the previous one year. A total of 2835 (39.9% of the total) practiced four days or more per week and 97.6% practiced 3 h or more per day on Saturdays and Sundays. The risk factors associated shoulder and elbow pain included a male sex, older age, pitchers and catchers, and players throwing more than 50 balls per day. Conclusions It has been revealed that Japanese elementary school baseball players train too much. Coaches should pay attention to older players, male players, pitchers and catchers in order to prevent shoulder and elbow pain. Furthermore, elementary school baseball players should not be allowed to throw more than 50 balls per day. Study design Retrospective cohort study.

A bizarre parosteal osteochondromatous proliferation (BPOP) is a rare, benign, osseous tumor that presents as an exophytic cortical growth consisting of bone, cartilage, and fibrous tissue. However, the identification of BPOP by preoperative imaging along with appropriate diagnosis and treatment without recurrence is challenging, particularly for BPOPs that rarely develop in certain parts of the body. Herein, we report a rare case involving a BPOP at the sesamoid bone of the hallux that was first suspected on preoperative computed tomography imaging. There has been no recurrence 3 years after surgical resection.

The application of fluorescent proteins in ornamental plants has lagged behind despite the recent development of powerful genetic tools. Although we previously generated transgenic torenia plants expressing green fluorescent protein from marine plankton (CpYGFP), in which bright fluorescence was easily visible at the whole plant level, the maximum excitation of this protein within the visible light spectrum required the use of a coloured emission filter to eliminate exciting light. Here, to overcome this limitation, we generated transgenic petunia plants expressing eYGFPuv, a CpYGFP derivative exhibiting bright fluorescence under invisible ultraviolet (UV) light excitation, with a novel combination of transcriptional terminator plus translational enhancer. As expected, all transgenic plants exhibited brilliant green fluorescence easily visible to the naked eye without an emission filter. In addition, fluorescence expressed in transgenic petunia flowers was stable during long-term vegetative propagation. Finally, we visually and quantitatively confirmed that transgenic petunia flowers resist to long-term exposure of UV without any damages such as fluorescence decay and withering. Thus, our whole-plant fluorescence imaging tool, that does not require high sensitive imaging equipment or special imaging conditions for observation, might be useful not only for basic plant research but also for ornamental purposes as a novel flower property.

Background The relationships between radiographic hallux valgus (HV) and various physical functions independent of knee osteoarthritis (KOA) were examined among residents of a mountain village in Japan. Methods Study participants were recruited from mountain village residents aged ≥50 years. Participants’ height, weight, and body mass index (BMI) were measured, and baseline data, including age, sex, and foot pain, were obtained using interviews and questionnaires. Radiography of the feet and knees was performed to assess the presence of HV (HV angle ≥20°) and KOA (Kellgren-Lawrence grade ≥ II). Grip strength, 6-m walk at usual and maximum speeds, single-leg stance time, and stand up from a chair time were evaluated as physical function performance tests. Plantar pressure patterns were also examined. Results Moderate-severe HV (HV angle ≥30 degrees), impaired grip strength and maximum walking speed, and painful HV reduced usual and maximum walking speeds independent of KOA. Hallux plantar pressure decreased according to the HV angle. Hallux plantar pressure was significantly lower in painful HV than in the no HV feet or painless HV. Conclusions Moderate-severe HV deformity and HV-related pain impaired physical function independent of KOA. By controlling the pain and severe deformity of HV by treatments such as surgery, the physical function of HV patients might be improved.

Background Locomotive syndrome (LS) is a condition of impaired mobility that is common in individuals with musculoskeletal disease who are at high risk of requiring nursing care. The "25-question Geriatric Locomotive Function Scale" (GLFS-25) was developed to help clinicians detect LS. The purpose of this study is to examine the relationship between the GLFS-25 score and knee osteoarthritis (KOA), osteoporosis (OP), and fall-related physical function in participants of a rural Japanese cohort study (Miyagawa study). Methods We examined 219 participants (>65 years old) of the 8 th Miyagawa study in 2011. All of the participants completed Kaigo Hoken and GLFS-25 questionnaires and underwent radiography of both knees for KOA, dual energy X-ray absorptiometry for OP, and physical function tests. We classified the participants with a GLFS-25 > 16 points into the LS group (versus the non-LS group). Results The LS group had more participants with KOA and OP as well as those who required nursing care by Kaigo Hoken than the non-LS group. Subjects in the LS group had worse physical function than those in the non-LS group. Conclusions The GLFS-25 questionnaire is considered a useful instrument because it accurately reflects KOA, OP, Kaigo Hoken nursing care need, and physical function.

We describe entrapment of the common peroneal nerve by a suture after surgical repair of the distal biceps femoris tendon. Complete rupture of the distal biceps femoris tendon of a 16-year-old male athlete was surgically repaired. Postoperative common peroneal nerve palsy was evident, but conservative treatment did not cause any neurological improvement. Reexploration revealed that the common peroneal nerve was entrapped by the surgical suture. Complete removal of the suture and external neurolysis significantly improved the palsy. The common peroneal nerve is prone to damage as a result of its close proximity to the biceps femoris tendon and it should be identified during surgical repair of a ruptured distal biceps femoris tendon.

The major obstacle of efficient transgene expression seems to be gene silencing, and one of the important factors in gene silencing is mRNA stability. Regulation of mRNA stability is an important aspect of the control of gene expression. mRNAs are degraded by both exonucleolytic digestion and endonucleolytic cleavage. However, with the exception of small RNA-guided cleavage, the mechanisms underlying endonucleolytic cleavage-dependent RNA degradation remain to be elucidated. High-throughput approaches for genome-wide profiling of RNA cleavage sites, collectively termed degradome sequencing, have been developed by several groups. These analyses have contributed to the identification of mRNA cleavage sites in plants, but due to selection of poly (A) mRNA in library preparation, these approaches cannot identify cleavage sites in a fully accurate manner. To address this issue, we developed a new experimental method, truncated RNA end sequencing (TREseq), which enabled us to accurately identify many cleavage sites. TREseq can also be used to estimate the efficiency of mRNA cleavage, revealing differences in base frequencies near cleavage sites that reflect differences in cleavage efficiency. These results will contribute to gain important knowledge about the stability of the transgene mRNA in the future.

Longitudinal flexor hallucis longus (FHL) tendon tears are sometimes complicated by posterior ankle impingement syndrome (PAIS), especially in ballet dancers. In recent years, PAIS has been treated endoscopically, but it is difficult to suture FHL tendon tears endoscopically. In this report, we describe how to suture the FHL tendon endoscopically with the Meniscal Viper Repair system (Arthrex, Naples, FL). Without our endoscopic technique, when a patient is found to have a longitudinal tear of the FHL under endoscopy, we must choose to either neglect the tear or convert to an open repair. Open tendon suture techniques have reportedly had relatively good results but require a longer skin incision than endoscopic surgery for PAIS. Compared with the open repair, the advantages of our technique include earlier recovery, less pain, a lower rate of soft tissue complications, and improved healing through better preservation of the blood supply. This technique is an attractive and useful option because it is an easy and safe method for longitudinal FHL tendon tears.

Transgene expression is regulated through several steps, this study focuses on the mRNA translation step. The expression level of transgenes can be increased by 5′-untranslated region (5′UTR) sequences in certain genes which act as translational enhancers. On the other hand, translation in most mRNA species is repressed by growth, development, and stress events. There is a possibility that transgene mRNA is also repressed in these conditions, despite the use of a translational enhancer. Therefore, a consistently efficient translational enhancer is needed to develop a reliable transgene expression system. Herein we searched for mRNAs translated stably under different growth, development and environmental conditions using data sets of polysome fraction assays and microarray analysis. Correct 5′UTR sequences of candidate genes were determined by cap analysis of gene expression and we tested translational ability of the candidate 5′UTRs by reporter assays. We found the 5′UTR of cold-regulated 47 gene to be an effective translational enhancer, contributing to stable high-level expression under various conditions.

The secondary cell walls of xylem cells, including vessel elements, provide mechanical strength and contribute to the conduction of water and minerals. VASCULAR-RELATED NAC-DOMAIN7 (VND7) is a NAC-domain transcription factor that regulates the expression of genes required for xylem vessel element formation. Transient expression assays using 68 transcription factors that are expressed during xylem vessel differentiation showed that 14 transcription factors, including VND1-VND7, are putative positive regulators of VND7 expression. Electrophoretic mobility shift assays revealed that all seven VND proteins bound to the VND7 promoter region at its SMBE/TERE motif, indicating that VND7 is a direct target of all of the VND transcription factors. Overexpression of VND1-VND5, GATA12 and ANAC075, newly identified transcription factors that function upstream of VND7, resulted in ectopic xylem vessel element formation. These data suggest that VND7 transcription is a regulatory target of multiple classes of transcription factors.

From Drips to Tubes In the evolutionary transition from aquatic to terrestrial habitats, plants acquired internal systems to transport water and provide structural support. Xu et al. (p. 1505 , published online 20 March) studied a family of genes and the cells they control to better understand the innovations required to adapt to dry land. In Arabidopsis , specific transcription factors regulate development of xylem—the plant tissue that transports water. The moss Physcomitrella patens has similar genes, which regulate development of hydroids and stereids, cells specialized in water transport and structural support. The similarity in the genes and their functions suggests the evolutionary origins of land-plant vascular systems.

Proximal humeral fractures are becoming increasingly common, and treatment of displaced proximal humeral fractures remains a challenge. Minimally invasive plate osteosynthesis (MIPO), which provides early functional recovery, is a promising treatment option for proximal humeral fractures [1]. We report a patient with posterior fracture–dislocation of the shoulder who was successfully treated with arthroscopic reduction and plate fixation via a minimally invasive approach.

In order to achieve higher expression of a transgene, it is important to optimize not only the promoter, 5′-untranslated region (UTR), and localization signal, but also the transcriptional terminator. We previously reported that the terminator derived from the Arabidopsis thaliana heat shock protein 18.2 gene (HSPT) increases gene expression in various plant species. In this study, with the goal of further increasing the expression level of transgenes, we evaluated a longer version of HSPT, corresponding to the 878 bp downstream of the stop codon, in cultured tobacco cells. The longer version of HSPT increased the expression levels of various genes including Renilla reniformis luciferase, Photinus pyralis luciferase, horseradish (Armoracia rusticana) peroxidase, and the non-toxic B subunit of Stx2e, a candidate vaccine protein for pig edema disease. This effect was not observed in a transient expression system. This element represents a useful tool for expression of transgenes integrated into the nuclear genome in plant cells.