NAC domain transcription factors VNI2 and ATAF2 form protein complexes and regulate leaf senescence.

The NAM, ATAF1/2, and CUC2 (NAC) domain transcription factor VND-INTERACTING2 (VNI2) negatively regulates xylem vessel formation by interacting with another NAC domain transcription factor, VASCULAR-RELATED NAC-DOMAIN7 (VND7), a master regulator of xylem vessel formation. Here, we screened interacting proteins with VNI2 using yeast two-hybrid assay and isolated two NAC domain transcription factors, Arabidopsis thaliana ACTIVATION FACTOR 2 (ATAF2) and NAC DOMAIN CONTAINING PROTEIN 102 (ANAC102). A transient gene expression assay showed that ATAF2 upregulates the expression of genes involved in leaf senescence, and VNI2 effectively inhibits the transcriptional activation activity of ATAF2. vni2 mutants accelerate leaf senescence, whereas ataf2 mutants delay leaf senescence. In addition, the accelerated leaf senescence phenotype of the vni2 mutant is recovered by simultaneous mutation of ATAF2. Our findings strongly suggest that VNI2 interacts with and inhibits ATAF2, resulting in negatively regulating leaf senescence.

Knotless Tendoscopic Peroneal Retinaculum Repair Technique for Recurrent Peroneal Tendon Dislocation.

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.

Sequence features around cleavage sites are highly conserved among different species and a critical determinant for RNA cleavage position across eukaryotes.

RNA degradation is one of the critical steps 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 Cap analysis of gene expression (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.

Methods for detecting RNA degradation intermediates in plants.

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.

Is musculoskeletal pain related to locomotive syndrome even in young and middle-aged adults?

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.

Is locomotion training effective for middle-aged workers?

OBJECTIVES: Locomotion training (LT) consisting of single-leg standing and squatting was developed to help prevent locomotive syndrome (LS), and is typically used in older people. The objective of this study was to examine the effects of LT on young and middle-aged people. METHODS: This study was performed at two companies. Workers in company A engaged in LT five times/week for 1 year, whereas workers in company B did not. Baseline and follow-up checkups consisted of questionnaires and physical performance tests, including three kinds of locomotion tests. RESULTS: In total, 88 and 101 workers in companies A and B, respectively, met the inclusion criteria. LS stage, stand-up test results, and scores on a geriatric locomotive function scale significantly improved among workers in company A, but only stand-up test results significantly improved among workers in company B. Quadriceps power increased in company A, but did not change in company B. Especially, workers with LS in company A had more significant changes than those without LS and those in company B. CONCLUSIONS: The results of this longitudinal study suggest that LT is useful even for young and middle-aged workers. LT was especially more effective for workers than those without LS.

Repair of Acute Patellar Tendon Rupture Using an Internal Brace Technique.

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.

Lettuce-based production of an oral vaccine against porcine edema disease for the seed lot system.

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.

Feature selection for RNA cleavage efficiency at specific sites using the LASSO regression model in Arabidopsis thaliana.

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.

Arthroscopic Internal Brace Augmentation With Arthroscopic Modified Broström Operation for Chronic Ankle Instability.

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.

Changes in mRNA Degradation Efficiencies under Varying Conditions Are Regulated by Multiple Determinants in Arabidopsis thaliana.

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.

Arthroscopic Treatment of Primary Synovial Chondromatosis of the Subscapular Bursa: A Case Report.

INTRODUCTION: Synovial chondromatosis is a rare metaplastic disorder of the synovial membrane. Primary synovial chondromatosis of the shoulder joint is a rare localization and extra-articular localization around the shoulder is much less common. CASE REPORT: We report a rare case of a 13-year-old boy with primary synovial chondromatosis of the subscapular bursa. Computed tomography and magnetic resonance imaging showed that multiple cartilaginous loose bodies were found in the subscapular bursa and the glenohumeral joint. Arthroscopic removal of loose bodies and synovectomy of the subscapular bursa was performed through sublabral foramen. CONCLUSION: Pre-operative investigation for the precise location of the lesions was important for the planning the operative procedure. Arthroscopic removal of loose bodies and synovectomy was useful treatment options for synovial chondromatosis of the subscapular bursa.

Tendoscopic peroneal retinaculum repair for recurrent peroneal tendon dislocation enables earlier return to sports than the open procedure.

PURPOSE: 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. METHODS: 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. RESULTS: 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). CONCLUSIONS: 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. LEVEL OF EVIDENCE: III.

Locomotive syndrome testing in young and middle adulthood.

Background: Previous epidemiological studies on locomotive syndrome (LS) have been mainly conducted in retired elderly people. However, from the view point of its prevention, it is important to assess currently working younger people. The purpose of this study was to examine the status of the level of LS in young and middle-aged adult workers.Methods: This cross-sectional study was performed at four companies. We used the two-step test, stand-up test, and 25-question Geriatric Locomotive Function Scale (GLFS-25) as screening tools of LS. We also questioned the subjects for knee pain and low back pain.Results: A total of 715 workers participated in this study. The percentage of females with LS increased with age. However, the percentage of males with LS was approximately 20% in each age category. Two-step and stand-up test scores worsened in an age-dependent manner, although GLFS-25 scores varied independently of age. Knee pain, low back pain and physical activity were significantly related to LS-level.Conclusion: Our results suggest that there is some degree of LS even in young and middle-aged adults. Knee pain, low back pain and physical activity levels are key factors related to LS even in young and middle-aged adults.

Different Plant Species Have Common Sequence Features Related to mRNA Degradation Intermediates.

mRNA degradation is an important cellular mechanism involved in the control of gene expression. Several genome-wide profiling methods have been developed for detecting mRNA degradation in plants and animals. However, because many of these techniques use poly (A) mRNA for library preparation, degradation intermediates are often only detected near the 3'-ends of transcripts. Previously, we developed the Truncated RNA End Sequencing (TREseq) method using Arabidopsis thaliana, and demonstrated that this method ameliorates 3'-end bias. In analyses using TREseq, we observed G-rich sequences near the 5'-ends of degradation intermediates. However, this finding remained to be confirmed in other plant species. Hence, in this study, we conducted TREseq analyses in Lactuca sativa (lettuce), Oryza sativa (rice) and Rosa hybrida (rose). These species including A. thaliana were selected to encompass a diverse range in the angiosperm phylogeny. The results revealed similar sequence features near the 5'-ends of degradation intermediates, and involvement of translation process in all four species. In addition, homologous genes have similar efficiencies of mRNA degradation in different plants, suggesting that similar mechanisms of mRNA degradation are conserved across plant species. These strong sequence features were not observed in previous degradome analyses among different species in plants.

Affinity-based high-resolution analysis of DNA binding by VASCULAR-RELATED NAC-DOMAIN7 via fluorescence correlation spectroscopy.

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.

Spontaneous Osteonecrosis of the Tarsal Navicular: A Report of Two Cases.

Spontaneous osteonecrosis of the tarsal navicular, called the Mueller-Weiss syndrome, is an uncommon disease. Patients who are resistant to conservative treatment require operative treatment. However, there is no established operative treatment. Two cases of spontaneous osteonecrosis of the tarsal navicular with double (talonavicular and naviculocuneiform joints) arthrodeses with different locking plates are presented. Removal of necrotic areas from the tarsal navicular and replacement with autologous bone graft procured from the iliac crest followed by arthrodesis using a locking plate were performed. Case 1 was fixed with an LCP Distal Radius Plate (SYNTHES) and 6 2.4 mm locking screws. Case 2 was fixed with a Cervical Spine Locking Plate Variable Angle (SYNTHES) and 4 4.0 mm locking screws. Case 2 achieved solid fusion of the talonavicular-cuneiform joints, but case 1 resulted in nonunion of the talonavicular joint. This difference in internal fixation strength might have caused the difference in the results. Performance of double arthrodeses from the medial aspect using a locking plate is a reasonable operative procedure to treat spontaneous osteonecrosis of the tarsal navicular. Strong primary fixation using a thick plate with large-thread screws was important to obtain joint fusions.

A bizarre parosteal osteochondromatous proliferation at the sesamoid bone of the hallux: A case report.

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.

Generation of brilliant green fluorescent petunia plants by using a new and potent fluorescent protein transgene.

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.

Does hallux valgus impair physical function?

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.