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Eukaryotic translation is a complex process that involves the interplay of various translation factors to convert genetic information into a specific amino acid chain. According to an elegant model of eukaryotic translation initiation, the 3' poly(A) tail of an mRNA, which is occupied by poly(A)-binding proteins (PABPs), communicates with the 5'-cap bound by eIF4E to enhance translation. Although the circularization of mRNA resulting from the communication is widely understood, it has yet to be directly observed. To explore mRNA circularization in translation, we analyzed the level of colocalization of eIF4E, eIF4G, and PABP on individual mRNAs in polysomal and subpolysomal fractions using single polysome analysis. Our results show that the three tested proteins barely coexist in mRNA in either polysomal or subpolysomal fractions, implying that the closed-loop structure generated by the communication between eIF4E, eIF4G, and PAPB may be transient during translation.

The roles of fibronectin leucine-rich transmembrane protein 2 (FLRT2) in physiological and pathological processes are poorly known. Here, we identify a novel function of FLRT2 in preventing endothelial cell senescence and vascular aging. We found that FLRT2 expression was lower in cultured senescent endothelial cells as well as in aged rat and human vascular tissues. FLRT2 silencing in human endothelial cells induced senescence through mTORC2, but not mTORC1, AKT, and p53. We uncovered that FLRT2 directly associated with ITGB4 and thereby promoted ITGB4 phosphorylation, while inhibition of ITGB4 significantly mitigated the induction of senescence triggered by FLRT2 depletion. Importantly, FLRT2 silencing in mice promoted vascular aging and overexpression of FLRT2 rescued a premature vascular aging phenotype. We propose that FLRT2 could be targeted therapeutically to prevent senescence-associated vascular aging. Subject terms: FLRT2, ITGB4, mTORC2, endothelial cell senescence, vascular aging

Newly synthesized mRNA is translated during its export through the nuclear pore complex, when its 5′-cap structure is still bound by the nuclear cap-binding complex (CBC), a heterodimer of cap-binding protein (CBP) 80 and CBP20. Despite its critical role in mRNA surveillance, the mechanism by which CBC-dependent translation (CT) is regulated remains unknown. Here, we demonstrate that the CT initiation factor (CTIF) is tethered in a translationally incompetent manner to the perinuclear region by the DEAD-box helicase 19B (DDX19B). DDX19B hands over CTIF to CBP80, which is associated with the 5′-cap of a newly exported mRNA. The resulting CBP80–CTIF complex then initiates CT in the perinuclear region. We also show that impeding the interaction between CTIF and DDX19B leads to uncontrolled CT throughout the cytosol, consequently dysregulating nonsense-mediated mRNA decay. Altogether, our data provide molecular evidence supporting the importance of tight control of local translation in the perinuclear region.

The hallmark of cellular events observed upon macroautophagic/autophagic induction is the conjugation of LC3B, one of the mammalian Atg8 homologs, with phosphatidylethanolamine. This conversion from LC3B-I (an unconjugated form) to LC3B-II (a conjugated form) is essential for phagophore expansion and formation of autophagosomes. Our recent study revealed that LC3B binds to RNAs with a preference for the consensus AAUAAA motif and recruits the CCR4-NOT deadenylase complex. Consequently, LC3B elicits rapid degradation of mRNAs, which we have termed as LC3B-mediated mRNA decay (LMD). LMD requires the conversion of LC3B-I to LC3B-II and occurs before the formation of autolysosomes. Furthermore, we identified

Newly synthesized mRNAs are exported from the nucleus to cytoplasm with a 5′-cap structure bound by the nuclear cap-binding complex (CBC). During or after export, the CBC should be properly replaced by cytoplasmic cap-binding protein eIF4E for efficient protein synthesis. Nonetheless, little is known about how the replacement takes place. Here, we show that double-stranded RNA-binding protein staufen1 (STAU1) promotes efficient replacement by facilitating an association between the CBC–importin α complex and importin β. Our transcriptome-wide analyses and artificial tethering experiments also reveal that the replacement occurs more efficiently when an mRNA associates with STAU1. This event is inhibited by a key nonsense-mediated mRNA decay factor, UPF1, which directly interacts with STAU1. Furthermore, we find that cellular apoptosis that is induced by ionizing radiation is accompanied by inhibition of the replacement via increased association between STAU1 and hyperphosphorylated UPF1. Altogether, our data highlight the functional importance of STAU1 and UPF1 in the course of the replacement of the CBC by eIF4E, adding a previously unappreciated layer of post-transcriptional gene regulation.

Aging causes a progressive decline in the structure and function of organs. With advancing age, an accumulation of senescent endothelial cells (ECs) contributes to the risk of developing vascular dysfunction and cardiovascular diseases, including hypertension, diabetes, atherosclerosis, and neurodegeneration. Senescent ECs undergo phenotypic changes that alter the pattern of expressed proteins, as well as their morphologies and functions, and have been linked to vascular impairments, such as aortic stiffness, enhanced inflammation, and dysregulated vascular tone. Numerous molecules and pathways, including sirtuins, Klotho, RAAS, IGFBP, NRF2, and mTOR, have been implicated in promoting EC senescence. This review summarizes the molecular players and signaling pathways driving EC senescence and identifies targets with possible therapeutic value in age-related vascular diseases.

Selective recognition and removal of faulty transcripts and misfolded polypeptides are crucial for cell viability. In eukaryotic cells, nonsense-mediated mRNA decay (NMD) constitutes an mRNA surveillance pathway for sensing and degrading aberrant transcripts harboring premature termination codons (PTCs). NMD functions also as a post-transcriptional gene regulatory mechanism by downregulating naturally occurring mRNAs. As NMD is activated only after a ribosome reaches a PTC, PTC-containing mRNAs inevitably produce truncated and potentially misfolded polypeptides as byproducts. To cope with the emergence of misfolded polypeptides, eukaryotic cells have evolved sophisticated mechanisms such as chaperone-mediated protein refolding, rapid degradation of misfolded polypeptides through the ubiquitin–proteasome system, and sequestration of misfolded polypeptides to the aggresome for autophagy-mediated degradation. In this review, we discuss how UPF1, a key NMD factor, contributes to the selective removal of faulty transcripts via NMD at the molecular level. We then highlight recent advances on UPF1-mediated communication between mRNA surveillance and protein quality control.

Loss of PTEN, the major negative regulator of the PI3K/AKT pathway induces a cellular senescence as a failsafe mechanism to defend against tumorigenesis, which is called PTEN-loss-induced cellular senescence (PICS). Although many studies have indicated that the mTOR pathway plays a critical role in cellular senescence, the exact functions of mTORC1 and mTORC2 in PICS are not well understood. In this study, we show that mTOR acts as a critical relay molecule downstream of PI3K/AKT and upstream of p53 in PICS. We found that PTEN depletion induces cellular senescence via p53-p21 signaling without triggering DNA damage response. mTOR kinase, a major component of mTORC1 and mTORC2, directly binds p53 and phosphorylates it at serine 15. mTORC1 and mTORC2 compete with MDM2 and increase the stability of p53 to induce cellular senescence via accumulation of the cell cycle inhibitor, p21. In embryonic fibroblasts of PTEN-knockout mice, PTEN deficiency also induces mTORC1 and mTORC2 to bind to p53 instead of MDM2, leading to cellular senescence. These results collectively demonstrate for the first time that mTOR plays a critical role in switching cells from proliferation signaling to senescence signaling via a direct link between the growth-promoting activity of AKT and the growth-suppressing activity of p53.

BACKGROUND: Quantitative assessment of rotator cuff muscle activity is important in the treatment of shoulder disorders. However, the known methods for assessing rotator cuff muscle activity thus far have been inaccurate, invasive, and inconvenient. QUESTIONS/PURPOSES: (1) Does the activity of the deltoid, supraspinatus, and infraspinatus muscles measured using ultrasound shear wave elastography have a linear correlation with muscle activity assessed using generally used methods, including isokinetic dynamometry and electromyography? (2) Does the activity of the deltoid, supraspinatus, and infraspinatus muscles measured using shear wave elastography show good intraobserver and interobserver reliability? METHODS: Twelve volunteers participated in intrasession reliability experiments. They were asked to perform isometric abduction, external rotation, and scaption contractions (defined as elevation of the arm within the plane of the scapula with neutral arm rotation) gradually increased from 0% to 75% of maximal voluntary contraction. The joint torque, electromyographic activity, and shear elastic modulus were synchronously measured in the middeltoid, supraspinatus, and infraspinatus muscles. The validity of the elastic modulus value was assessed using regression analysis between normalized torque and electromyographic root mean square values. For intraobserver and interobserver reliability measurements, repeated experiments were performed with the same protocol. RESULTS: The shear elastic modulus and normalized joint torque with isokinetic dynamometry showed a linear relationship in all muscles (deltoid, supraspinatus, and infraspinatus) and each of the ultrasonography planes (longitudinal and transverse) (mean R(2) > 0.8 and p < 0.001 for all measurements). For the supraspinatus muscle, the mean slope of the relationship between shear elastic modulus in the longitudinal plane and the normalized joint torque during scaption contraction was 1.28 ± 0.39 kPa/%MVC (mean R(2) = 0.93 ± 0.21, p < 0.001). Furthermore, similar results were obtained in relation to electromyography root mean square values (mean R(2) > 0.8 and p < 0.001 in all measurements). For the supraspinatus muscle, the mean slope of the relationship between shear elastic modulus in the longitudinal plane and electromyographic (EMG) root mean square was 0.96 ± 0.27 kPa/%EMG (mean R(2) = 0.91 ± 0.08, p < 0.001). The intraobserver and interobserver reliabilities were excellent in all positions (abduction, external rotation, and scaption) and in both the longitudinal and transverse ultrasonography planes (all intraclass correlation coefficients are > 0.85). CONCLUSIONS: Shoulder muscle activity can be noninvasively evaluated with ultrasound shear wave elastography. Clinician and scientists should consider the application of this technique in cases in which evaluation of shoulder muscle activity is required. The next step after this study will be to check the shear elastic modulus of rotator cuff muscle in patients with rotator cuff tear. We plan to evaluate the correlation between shear elastic modulus and joint torque according to tear size and fatty infiltration status of rotator cuff muscle. CLINICAL RELEVANCE: Shear wave electrography can be used to measure various tissue elasticities in both static and dynamic modes. It may be a useful tool to evaluate pre- and postoperative rotator cuff muscle activity in a relatively simple manner. Shoulder function after reverse total shoulder arthroplasty associated with deltoid muscle activity also may be evaluated. Changes in tissue tightness in shoulder disorders caused by increase soft tissue stiffness (ie, adhesive capsulitis and glenohumeral internal rotation deficit) can be evaluated.

The effects of senescence associated secretory phenotype (SASP) from therapy-induced senescent endothelial cells on tumor microenvironment (TME) remains to be clarified. Here, we investigated effects of ionizing radiation (IR)- and doxorubicin-induced senescent HUVEC on TME. MDA-MB-231 cancer cells treated with conditioned medium (CM) from senescent HUVEC or co-cultured with senescent HUVEC significantly increased cancer cell proliferation, migration, and invasion. We found that CXCL11 plays a principal role in the senescent CM-induced aggressive activities of MDA-MB-231 cells. When we treated HUVEC with a neutralizing anti-CXCL11 antibody or CXCL11 SiRNA, or treated MDA-MB-231 cells with CXCR3 SiRNA, we observed synergistic diminution of the ability of the HUVEC SASP to alter the migration and spheroid invasion of cancer cells. ERK activation was involved in the HUVEC SASP-induced aggressive activity of MDA-MB-231 cells. Finally, we observed the in vivo effect of CXCL11 from the senescent HUVEC in tumor-bearing mice. Together, our results demonstrate that SASP from endothelial cells experiencing therapy-induced senescence promotes the aggressive behavior of cancer cells, and that CXCL11 can potentially be targeted to prevent the adverse effects of therapy-induced senescent endothelial cells on the tumor microenvironment.

Long noncoding RNAs (lncRNAs) regulating diverse cellular processes implicate in many diseases. However, the function of lncRNAs in cellular senescence remains largely unknown. Here we identify a novel long intergenic noncoding RNA Linc-ASEN expresses in prematurely senescent cells. We find that Linc-ASEN associates with UPF1 by RNA pulldown mass spectrometry analysis, and represses cellular senescence by reducing p21 production transcriptionally and posttranscriptionally. Mechanistically, the Linc-ASEN-UPF1 complex suppressed p21 transcription by recruiting Polycomb Repressive Complex 1 (PRC1) and PRC2 to the p21 locus, and thereby preventing binding of the transcriptional activator p53 on the p21 promoter through histone modification. In addition, the Linc-ASEN-UPF1 complex repressed p21 expression posttranscriptionally by enhancing p21 mRNA decay in association with DCP1A. Accordingly, Linc-ASEN levels were found to correlate inversely with p21 mRNA levels in tumors from patient-derived mouse xenograft, in various human cancer tissues, and in aged mice tissues. Our results reveal that Linc-ASEN prevents cellular senescence by reducing the transcription and stability of p21 mRNA in concert with UPF1, and suggest that Linc-ASEN might be a potential therapeutic target in processes influenced by senescence, including cancer.

The purpose of this study was to evaluate changes in the quality characteristics of mackerel fish fillet under freezing storage conditions. The changes in product quality were determined by measuring pH, volatile basic nitrogen (VBN); thiobarbituric acid (TBA), lightness (L * ), redness (a * ), yellowness (b * ) values, shearing force, and microbiological evaluation (Total Plate Count, TPC) during 21 days of storage at −20, −5, and −1℃, and their shelf-lives were established. TPC as an effective quality indicator was used to estimate the shelf-life via linear regression analysis. The TPC increased steadily with increasing storage periods at every temperature (p

// Beom-Chan Kim 1, 2 , Hyun-Jung Hwang 1, 3 , Hyoung-Tae An 1, 2 , Hyun Lee 1, 2 , Jun-Sub Park 1, 2 , Jin Hong 1, 2 , Jesang Ko 1, 2 , Chungho Kim 1 , Jae-Seon Lee 3 , Young-Gyu Ko 1, 2 1 Tunneling Nanotube Research Center, Korea University, Seoul, 02841, Korea 2 Division of Life Sciences, Korea University, Seoul, 02841, Korea 3 Department of Molecular Medicine, College of Medicine, Inha University, Incheon, 22212, Korea Correspondence to: Young-Gyu Ko, email: ygko@korea.ac.kr Keywords: gC1qR, lamellipodia, cell migration, antibody, cancer Received: January 07, 2016 Accepted: May 28, 2016 Published: June 24, 2016 ABSTRACT We previously demonstrated that cell-surface gC1qR is a key regulator of lamellipodia formation and cancer metastasis. Here, we screened a monoclonal mouse antibody against gC1qR to prevent cell migration by neutralizing cell-surface gC1qR. The anti-gC1qR antibody prevented growth factor-stimulated lamellipodia formation, cell migration and focal adhesion kinase activation by inactivating receptor tyrosine kinases (RTKs) in various cancer cells such as A549, MDA-MB-231, MCF7 and HeLa cells. The antibody neutralization of cell-surface gC1qR also inhibited angiogenesis because the anti-gC1qR antibody prevented growth factor-stimulated RTK activation, lamellipodia formation, cell migration and tube formation in HUVEC. In addition, we found that A549 tumorigenesis was reduced in a xenograft mouse model by following the administration of the anti-gC1qR antibody. With these data, we can conclude that the antibody neutralization of cell-surface gC1qR could be a good therapeutic strategy for cancer treatment.

Osteomyelitis originating in the epiphysis of the long bones is quite rare and is usually found at either the distal femur or the proximal tibia. We report the case of a 12-year-old male with epiphyseal osteomyelitis that had developed in the distal tibia. To the best of our knowledge, this is the first published case report. The patient's history of a trauma that resembled an ankle sprain had delayed the diagnosis and subsequently led him to develop septic arthritis. The ankle is a common site of simple trauma; however, epiphyseal osteomyelitis is rare at this site. Therefore, if the symptoms continue or worsen after trauma, the clinician should check the affected site and take a more aggressive approach to make an early diagnosis.

Cellular senescence refers to an irreversible growth arrest that is triggered by various intrinsic and extrinsic stresses. Many recent studies have demonstrated that cellular senescence plays a crucial role in the regression of tumors exposed to ionizing radiation (IR), but the underlying mechanism remains unknown. Here we show that the activation of integrin β4 is essential for IR-induced cellular senescence. IR treatment results in the phosphorylation of integrin β4 at tyrosine residue 1510, leading to activation of the integrin α6β4-Src-AKT signaling pathway. We further reveal that the IR-induced phosphorylation of integrin β4 is regulated by the cholesterol content and membrane fluidity. We also find that IR-induced p53-caspase signaling is independent of integrin α6β4-Src-AKT signaling. Finally, we show that siRNA- or inhibitor-mediated blockade of integrin α6β4-Src-AKT signaling switches the post-irradiation fate from senescence to apoptosis, under p53 activated condition, in both cancer cells and tumor tissues of xenograft mice. On the basis of our finding that, integrin α6β4 is specifically activated and acts primarily to induce premature senescence in irradiated cancer cells, we propose that this integrin may be a valuable target and biomarker for radiotherapy.

This study was conducted to establish roasting conditions for optimization of Citri Unshii Pericarpium antioxidant activity using response surface methodology (RSM). A central composite design was applied to investigate the effects of two independent variables, namely roasting temperature (40∼100°C; X1) and roasting time (5∼15 min; X2), on responses such as electron donating ability (Y1), total phenolic content (Y2), total flavonoid content (Y3), and hydroxyl radical scavenging activity (Y4). The maximum electron donating ability was 72.38% at a roasting temperature of 71.12°C and roasting time of 9.39 min. The maximum total phenolic content was 10.76 mg tannic acid equivalents/g at a roasting temperature of 69.71°C and roasting time of 8.39 min. The maximum total flavonoid content was 105.99 mg quercetin equivalents/100 g at 72.54°C and 8.64 min. The maximum hydroxyl radical scavenging activity was 60.33% at 68.97°C and 9.84 min. Based on the superimposition of three dimensional RSM with respect to electron donating ability, total phenolic content, total flavonoid content, and hydroxyl radical scavenging activity under various conditions, optimum conditions were established as follows: roasting temperature of 70.90°C and roasting time of 9.03 min.

Cellular senescence is a state of irreversible growth arrest that can be triggered by multiple mechanisms, including telomere shortening, the epigenetic derepression of the INK4α/ARF locus and DNA damage. Senescence has been considered a tumor‑suppressing mechanism that permanently arrests cells at risk for malignant transformation. However, accumulating evidence shows that senescent cells have deleterious effects on the tissue microenvironment. Some of these effects could be attributed to the senescence‑associated secretory phenotype that has the ability to promote tumor progression. However, secreted proteins from senescent tumor cells and their effects on the tumor microenvironment due to ionizing radiation (IR) exposure have not yet been fully elucidated. In the present study, we analyzed cytokines secreted from IR‑induced senescent MCF7 cells by using cytokine microarrays and confirmed by western blot analysis that increased secretion of osteoprotegerin (OPG), midkine (MDK) and apolipoprotein E3 (ApoE3) occurs in these cells. Invasive, migratory and wound‑healing activities were observed in MDA‑MB‑231 and MCF‑10A cells following treatment with recombinant human OPG, MDK and ApoE3 proteins. Additionally, tube‑formation activity was assessed in OPG‑, MDK‑ and ApoE3‑treated human umbilical vein endothelial cells (HUVECs). We found that OPG, MDK and ApoE3 affected cell motility and tube‑formation activity. Since OPG markedly affected cell motility, we examined the effect of senescent conditioned media containing neutralizing OPG antibodies on migration and wound‑healing activity. Our results demonstrated that IR‑induced senescent tumor cells influence the tumor microenvironment by increasing the production of cytokines, such as OPG, MDK and ApoE3. Furthermore, these data suggest that OPG is likely a promising target capable of reducing the deleterious effects on the tumor microenvironment during radiation therapy.

Background Numerous authors have evaluated the strength of the rotator cuff muscles in patients with shoulder instability. However, only limited data are available with regard to neuromuscular control in patients with traumatic anterior shoulder instability, in particular at 90° of abduction. This study was designed to assess muscle strength and neuromuscular control ability using time to peak torque and acceleration time in nonathletic patients with traumatic anterior shoulder instability. Methods Isokinetic muscle performance testing was performed in 20 male nonathletic anterior shoulder instability patients compared with 20 side-matched asymptomatic volunteers. Isokinetic muscle performance testing was performed at an angular velocity of 180°/s with 90° of shoulder abduction. Muscle strength and neuromuscular control (time to peak torque and acceleration time) of the internal rotators (IRs) and external rotators (ERs) were measured. Results There were no significant differences in muscle strength of the IRs and ERs between the 2 groups. The injured shoulder showed delayed neuromuscular control in both the IRs and ERs in the instability patients compared with the normal control subjects (time to peak torque, P = .023 for IRs and P = .020 for ERs; acceleration time, P = .035 for IRs and P = .021 for ERs). Conclusion The neuromuscular control of both the IRs and ERs was decreased in male nonathletic patients with traumatic anterior shoulder instability even though muscle strength was not altered. Therefore, clinicians and therapists should implement exercises that aim to restore neuromuscular control in the rehabilitation of nonathletic patients with anterior shoulder instability.

Essential oils of various plants have been known for potential biological effects such as antibacterial, antifungal, spasmolytic, antiplasmodial activities and insect-repellent property. Recently, the essential oils have attracted considerable interest in oral disease therapy. This essential oil has been known as being effective on easing sick house syndrome, giving forest aroma therapy effect and acting as repellent against pest. The essential oil of Pinus koraiensi, a native plant from Hongcheon-gun, Gangwon-do, was obtained by hydrodistillation. In light of its medicinal importance, in this study its composition, antibacterial activity and the reducing effect of offensive odor have been analyzed. The composition of essential oil was determined by GC and GC-MS. We have identified 14 compounds, of which 1R--pinene (19.38 %), 3-carene (10.21 %), camphene (9.82 %), limonene (9.00 %), bicyclo[2,2,1] heptan-2-ol (8.76 %) and -phellandrene (7.98 %) were the main components. Essential oils from P. koraiensis, Chamaecyparis obtusa, Abies holophylla and Pinus densiflora were compared in terms of alleviating effect of malodors caused from formaldehyde, ammonia, trimethylamine and methylmercaptan. P. koraiensis essential oil was found to decrease the amounts of ammonia and trimethylamine by 75.17 % and 77.36 %, respectively. Antibacterial activity against Streptococcus mutans and Streptococcus sobrinus, which were known as oral cavity inducer, was investigated using the paper disc agar diffusion method. The inhibition zone was observed against S. mutans (5.97 mm) and S. sobrinus (1.40 mm), respectively. P. koraiensis essential oil shown effective deodorization and inhibitory activity against oral cavity in this study might be potential material in oral sanitary industry.

Traditionally, Codonopsis lanceolata root have been used as a source of natural heath food. This study was initiated to investigate the impacts of methyl jasmonate (MeJA) and salicylic acid (SA) on adventitious growth C. lanceolata , the production of secondary metabolites, such as flavonoids, total phenolic compound, antioxidative activity (DPPH). The highest phenolics content was observed in treatment of 20 uM MeJA (74.53 mg/g). The content of total flavonoids followed the similar pattern as that of total phenolics, showing 38.45 mg/g of C. lanceolata treated by 20 uM MeJA. The DPPH scavenging activity was 24.2 (IC 50 ) of C. lanceolata treated by 20 uM MeJA. These results provide useful information for enhancing biological properties of cultural roots of C. lanceolata. Keywords Antioxidative, Codonopsis lanceolata , Methyl jasmonate, Salicylic acid, Secondary metabolite 서 론 더덕( Codonopsis lanceolata )은 초롱꽃과(Campanulaceae)의 숙근성(perennial plant) 다년생 식물로서 뿌리는 도라지( Platycodon grandiflorum

RNA-binding proteins (RBPs) are involved in mRNA splicing, maturation, transport, translation, storage and turnover. Here, we identified ACOT7 mRNA as a novel target of human WIG1. ACOT7 mRNA decay was triggered by the microRNA miR-9 in a WIG1-dependent manner via classic recruitment of Argonaute 2 (AGO2). Interestingly, AGO2 was also recruited to ACOT7 mRNA in a WIG1-dependent manner in the absence of miR-9, which indicates an alternative model whereby WIG1 controls AGO2-mediated gene silencing. The WIG1–AGO2 complex attenuated translation initiation via an interaction with translation initiation factor 5B (eIF5B). These results were confirmed using a WIG1 tethering system based on the MS2 bacteriophage coat protein and a reporter construct containing an MS2-binding site, and by immunoprecipitation of WIG1 and detection of WIG1-associated proteins using liquid chromatography-tandem mass spectrometry. We also identified WIG1-binding motifs using photoactivatable ribonucleoside-enhanced crosslinking and immunoprecipitation analyses. Altogether, our data indicate that WIG1 governs the miRNA-dependent and the miRNA-independent recruitment of AGO2 to lower the stability of and suppress the translation of ACOT7 mRNA.

To help reduce high intake of sodium in the Korean diet, sodium-reduced fried fish cakes (SRFFCs) were prepared and evaluated with regard to color, textural properties, and sensory attributes as indicators of quality. The quality characteristics of 30% SRFFCs were not notably different from those of full sodium FFCs; however, substitution of sodium with potassium altered the color and decreased consumer acceptance on sensory evaluation items. These results suggest that the SRFFCs that will be accepted by consumers can be prepared without compromising the quality.

Postmenopausal women are at an increased risk of developing coronary artery disease, primarily due to dyslipidemia that accompanies the loss of estrogen secretion. This study was performed to investigate the effects of Opuntia humifusa seed extracts on blood flow and serum lipid level in ovariectomized rats. Twenty-eight 9-week old female Sprague-Dawley rats were randomly assigned to four groups as sham-operated rats (SHAM), ovariectomized rats (OVX-CON), and ovariectomized rats that were treated with Opuntia humifusa seed extracts (OVX-OHS 2% and OVX-OHS 6%). The diets were fed to the rats for 7 weeks after operation. Serum total cholesterol and triglyceride contents decreased in the SHAM group compared to the OVX-CON group. Seven weeks of feeding of Opuntia humifusa seed extracts resulted in significant (p

In this study, the effect of the Opuntiahumifusa extracts on proliferation, alkaline phosphatase (ALP) activity, collagen synthesis and ROS level of a cell was investigated using an osteoblast. Opuntiahumifusawas separated intoOpuntiahumifusapeel (OH-P), seed (OH-Se) and stem (OH-St).These were subjected to extraction by using hot water and ethanol. The proliferation of the MC3T3-E1 osteoblastic cells that were treated with OH-Se water extract were increased by approximately 120%. Regarding the effects of OH-Se on ALP activity, the 50 μg/ml ethanol extract group showed the highest activity. The synthesis of collagen increased significantly in response to treatment with OH-Se water extract. The ROS scavenging effects of Opuntiahumifusawere investigated for involvement of oxidativedamage, cell culture and staining. Also, when OH-Se water extract 100 μg/ml was added, the ROS level decreased by 54%. These results indicate that Opuntiahumifusa extracts have an anabolic effect on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases.

Pine needles have long been used as a traditional health-promoting medicinal food in Korea. This study was carried out to investigate the effects of pine needle extracts on the antioxidant activity, and proliferation of osteoclastic RAW 264.7 cells. Pine needle extracts were examined using hot water, ethanol, hexane, hot water-ethanol, and hot water-hexane. The effects of the pine needle extracts were examined by comparing the results with that of a commercial agents, proanthocyanidin. Analysis of each extract indicated that hot water-ethanol and ethanol extracts contained the highest total polyphenol concentrations. The hot water-ethanol and ethanol extracts also showed relatively the highest SOD-like activity. The proliferation of osteoclastic RAW 264.7 cells treated with pine needle extracts was decreased by lower than 70%. In addition, the hot water and ethanol extracts of pine needle significantly reduced the number of tartrate-resistant acid phosphatase-positive () multinucleated cells from osteoclatic RAW 264.7 cells. These results indicate that pine needle extracts had an anabolic effect on bone through the promotion of osteoclast differentiation, suggesting that they could be used for the treatment of common metabolic bone diseases.

Pine (pinus densiflora) needles have long been used as a traditional health-promoting medicinal food in Korea. This study was conducted to investigate the effects of pine needle extracts on the hepatic antioxidant system in the damaged liver of carbon tetrachloride (CCl₄)-treated rats. Nine-week-old Sprague Dawley rats were divided into four groups: normal group (NOR), CCl₄-treated group (CCL), pine needle hot water extract and CCl₄-treated group (CCL-P), and Vitamin C and CCl₄-treated group (CCL-V). The enzyme activities and antioxidant effects of the pine needle hot water extracts were investigated at the levels of liver homogenates and serum of rats intoxicated with CCl₄. Serum GOT and GPT activities by CCl₄ treatment increased compared to those of the NOR group. However, they tended to decrease in the hot water extract-administered group. Liver SOD activity in the CCL group was significantly lower than the NOR group (p＜0.05). However, they increased in the CCL-P group compared to the CCL group. Further, the CAT and GPx activities of serum treated with CCl₄ were higher compared to those of the NOR group but lower in the CCL-P group compared to CCL group. These results suggest that pine needle hot water extract increases antioxidant activities.

Estrogen deficiency in peri- and postmenopausal women results in variety of neurovegetative, psychic and somatic symptoms, and may contribute to severe diseases within the aged female population. Ecklonia stolonifera (ES) is an edible brown algae traditionally used in fishery towns in Far East Asia. This study was performed to investigate the effects of ES extracts on blood flow and serum lipid concentration in ovariectomized rats. Weight-matched female Sprague-Dawley strain rats were assigned to four groups. Three groups were surgically ovariectomized (OVX). The fourth group was sham operated. Rats were randomly assigned to the following groups: sham-operated rats (Sham), ovariectomized control rats (OVX-CON), ovariectomized rats supplemented with ES extract at 50 mg/kg bw/day (OVX-ES50) and ovariectomized rats supplemented with ES extract at 200 mg/kg bw/day (OVX-ES200). Serum total cholesterol and triglyceride contents decreased in the sham group compared to the OVX-CON group. Six weeks feeding of ES extract resulted in a significant (p

Osteoporosis is a disease involving a decrease in bone mineral density and an increased risk of fractures. The MC3T3-E1 pre-osteoblastic cell line is a well-accepted model of osteogenesis in vitro. Pine needles have long been used as a traditional health-promoting medicinal food in Korea. In this study, MTT assay, the alkaline phosphatase (ALP) activity and collagen synthesis of osteoblast cells were investigated to determine the effects of pine needle extracts on cell proliferation and differentiation. Pine needle extracts were prepared using hexane, ethanol and water. The effects of the pine needle extracts were examined by comparing the results with those of commercial agents, such as proanthocyanidin. The MOT3-E1 cells exposed to proanthocyanidin showed increased proliferation in a concentration-dependent manner. The cells exposed to the hexane extract showed a similar increase in proliferation to that observed with proanthocyanidin. The hexane extract showed the highest ALP activity. Moreover, a supplement of pine needle extracts induced collagen synthesis in MC3T3-E1 cells. The pine needle extract produced the highest level of collagen synthesis at concentrations of 10~50 ㎍/㎖. These results indicate that pine needle extracts have an anabolic effect on bone by promoting osteoblastic differentiation, and may be used in the treatment of common metabolic bone diseases.

Prostaglandin E(2) (PGE(2)) has a strong protective effect on the gastric mucosa in vivo; however, the molecular mechanism of a direct cytoprotective effect of PGE(2) on gastric mucosal cells has yet to be elucidated. Although we reported previously that PGE(2) inhibited gastric irritant-induced apoptotic DNA fragmentation in primary cultures of guinea pig gastric mucosal cells, we show here that PGE(2) inhibits the ethanol-dependent release of cytochrome c from mitochondria. Of the four main subtypes of PGE(2) receptors, we also demonstrated, using subtype-specific agonists, that EP(2) and EP(4) receptors are involved in the PGE(2)-mediated protection of gastric mucosal cells from ethanol-induced apoptosis. Activation of EP(2) and EP(4) receptors is coupled with an increase in cAMP, for which a cAMP analogue was found here to inhibit the ethanol-induced apoptosis. The increase in cAMP is known to activate both protein kinase A (PKA) and phosphatidylinositol 3-kinase pathways. An inhibitor of PKA but not of phosphatidylinositol 3-kinase blocked the PGE(2)-mediated protection of cells from ethanol-induced apoptosis, suggesting that a PKA pathway is mainly responsible for the PGE(2)-mediated inhibition of apoptosis. Based on these results, we considered that PGE(2) inhibited gastric irritant-induced apoptosis in gastric mucosal cells via induction of an increase in cAMP and activation of PKA, and that this effect was involved in the PGE(2)-mediated protection of the gastric mucosa from gastric irritants in vivo.

Acyl-CoA thioesterase 7 (ACOT7) is a major isoform of the ACOT family that catalyzes hydrolysis of fatty acyl-CoAs to free fatty acids and CoA-SH. However, canonical and non-canonical functions of ACOT7 remain to be discovered. In this study, for the first time, ACOT7 was shown to be responsive to genotoxic stresses such as ionizing radiation (IR) and the anti-cancer drug doxorubicin in time- and dose-dependent manners. ACOT7 knockdown induced cytostasis via activation of the p53–p21 signaling pathway without a DNA damage response. PKCζ was specifically involved in ACOT7 depletion-mediated cell cycle arrest as an upstream molecule of the p53–p21 signaling pathway in MCF7 human breast carcinoma and A549 human lung carcinoma cells. Of the other members of the ACOT family, including ACOT1, 4, 8, 9, 11, 12, and 13 that were expressed in human, ACOT4, 8, and 12 were responsive to genotoxic stresses. However, none of those had a role in cytostasis via activation of the PKCζ–p53–p21 signaling pathway. Analysis of the ACOT7 prognostic value revealed that low ACOT7 levels prolonged overall survival periods in breast and lung cancer patients. Furthermore, ACOT7 mRNA levels were higher in lung cancer patient tissues compared to normal tissues. We also observed a synergistic effect of ACOT7 depletion in combination with either IR or doxorubicin on cell proliferation in breast and lung cancer cells. Together, our data suggest that a low level of ACOT7 may be involved, at least in part, in the prevention of human breast and lung cancer development via regulation of cell cycle progression.

A 34-year-old male presented with right knee instability without any trauma. He had been diagnosed with right knee gouty arthritis 2 years prior. An arthroscopic examination revealed abundant calcific material deposited around the knee joint, including in the ACL tissue, and that the ACL was torn at the femoral attachment site. Treatment involved a synovectomy to remove calcific material, followed by an ACL reconstruction. Histology evaluation revealed gouty arthritis with the presence of tophi in the synovium, soft tissue, and ACL tissue. The case presented here indicates the possibility of pathologic rupture of the ACL associated with gouty tophus infiltration of that ligament. Level of evidence IV.

Full article available online at OrthoSuperSite.com/view.aspx?rID=79490 A woman presented with knee pain and locking. Pain was exacerbated at the end of the range of motion, especially during extension, with locking symptoms similar to those as- sociated with a meniscus bucket handle tear. Ligamentous laxity was not defi nite. Plain radiographs showed multiple calcifi ed loose bodies. Magnetic resonance imaging (MRI) showed a lobulated mass that was hypointense to muscle on T1-weighted sequences and hyperintense to muscle on T2-weighted sequences in the anterior cruciate ligament (ACL). Arthroscopically, multiple loose bodies were observed in the intercondylar notch and pos- terolateral compartment. A huge mass replaced the normal ACL and was caught in the intercondylar notch. The mass in the intercondylar notch caused loss of extension range of motion (ROM) because the piece caused a mechanical blockage. However, the loss of fl exion ROM was likely caused by a loss of elasticity of the ligament rather than mechani- cal blockage. We resected the ACL mass, and removed the free bodies from the postero- lateral corner. It was not possible to preserve the ACL fi bers. Histological examination confi rmed a diagnosis of osteochondromatosis. All symptoms resolved postoperatively. At 20 months postoperatively, the patient was pain free and had regained full knee motion without recurrence evidenced by follow-up MRI. However, ACL removal caused the knee instability. To date the patient has not undergone ACL reconstruction because she prefers conservative treatment and has experienced little discomfort in activities of daily living. To our knowledge, this is the fi rst report to describe synovial osteochondromatosis wholly replacing the ACL fi bers and causing mechanical blocking of both extension and fl exion.

In contrast to transport across basolateral membranes, the mechanism governing transport of organic anions across the luminal membranes of proximal tubules has remained unclear. We recently found Tetracycline transporter-like protein (TETRAN), a human ortholog of yeast Tpo1p that can transport anionic Non-steroidal anti-inflammatory drugs (NSAIDs). In this study, we examine the expression and function of TETRAN. TETRAN mRNA is expressed in various human tissues, including kidney. When overexpressed in cultured cells, TETRAN was predominantly localized on cytoplasmic membranes. Immunohistochemical analysis of human and mouse kidney tissue showed that TETRAN was expressed at the luminal membranes of proximal tubules. Overexpression of TETRAN in cultured cells facilitated the uptake of organic anions such as indomethacin (a NSAID) and fluorescein. The results suggest that TETRAN is a novel human organic anion transporter, and that it serves as a transporter for some NSAIDs and various other organic anions at the final excretion step.

Recombinant whole heavy chains (H, 100 kDa) and their N-terminal (Hn, 50 kDa) and C-terminal (Hc, 50 kDa) half fragments of Clostridium botulinum type C and D neurotoxins were expressed as glutathione S-transferase (GST) fusion proteins in Escherichia coli. GST eliminated-preparations of H (10 microg), Hn (5 microg), Hc (5 microg), or a mixture of Hn (5 microg) and Hc (5 microg) of types C and D were mixed with an equal volume of adjuvant, and then were twice injected into mice subcutaneously. After immunization, the mice were challenged with up to 10(6) the minimum lethal doses (MLD)/0.5 ml of C or D toxin, the type of which was same as that of the immunogens. All of the mice immunized with antigens except for Hn survived against 10(5) to 10(6) MLD/0.5 ml of the toxins, but the mice immunized with Hn were killed by 100 MLD/0.5 ml. The mice immunized with a mixture of C-Hc and D-Hc, each 5 microg, also showed a high level of resistance against both C and D toxins. Antibody levels immunized with GST fused-or GST eliminatedpreparation were quite similar. These results indicate that recombinant GST-fused Hc can be used as a safe and effective vaccine for type C and D botulism in animals. It also became clear that one time inoculation with a large amount of C-Hc or D-Hc, 100 microg, is useful for vaccine trials in mice.

Non-steroidal anti-inflammatory drugs (NSAIDs), such as indomethacin, have serious gastrointestinal side effects. Since their direct cytotoxicity was suggested to be involved in this side effect, we here tried to identify NSAID-resistant genes. We screened for Saccharomyces cerevisiae genes whose overexpression causes indomethacin resistance and identified the TPO1 gene, which encodes a major facilitator superfamily transporter. Its overexpression or deletion made yeast cells resistant or sensitive, respectively, to some NSAIDs. A BLAST search identified the possible human orthologue of Tpo1p, tetracycline transporter-like protein (TETRAN), whose overexpression in cultured human cells caused resistance to some NSAIDs, suggesting that TETRAN is an efflux pump for some NSAIDs.

The structural genes for strain C-Stockholm (c-st) phage particles, a representative type C toxin-converting phage of Clostridium botulinum, have been determined. First, by determining the N-terminal amino acid sequences of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) bands of c-st phage particles, it became clear that four proteins, 14, 25, 32 and 42 kDa, are the products of the ORFs, cst166, cst165, cst160 and cst164, respectively, of the c-st phage genome. The Western blot analyses reacting these phage bands with an antiphage serum prepared previously indicated that the products of cst165 and cst160 are the main proteins of the phage particles. Then, six candidates for the phage structural proteins, including cst165 and cst160 gene products, were prepared as recombinant proteins. Also, the protein corresponding to the cst164 gene product was excised from SDS-PAGE gels. The antibodies against these seven proteins were prepared in rabbits, and finally, the reaction of these antibodies to the c-st phage particles was analyzed by electron microscopy. It was concluded that a sheath protein and a head protein of the c-st phage are the products of genes cst160 and cst165, respectively, and that these two proteins are conserved in the other three converting phages, but not in the nonconverting phage.

Clostridium botulinumtype B strain produces two forms of progenitor toxin, 16S and 12S. The 12S toxin is formed by association of a neurotoxin (NTX) and a non-toxic non-haemagglutinin (NTNH), and the 16S toxin is formed by conjugation of the 12S toxin with a haemagglutinin (HA). HA consists of four subcomponents designated HA1, HA2, HA3a and HA3b. When mice were immunized with formalin-detoxified NTX, 12S or 16S, a significantly greater amount of anti-NTX antibody (Ab) was produced in the mice injected with 16S than in NTX- or 12S-injected mice. Immunization with NTX mixed with HA1 and/or HA3b also increased the anti-NTX Ab production, whereas NTX mixed with HA2 did not, indicating that HA1 and HA3b have adjuvant activity. This was further confirmed by immunizing mice with human albumin (Alb) alone or Alb mixed with either HA1 or HA3b. When mouse-spleen cells were stimulated with NTX, 16S or different HA subcomponents, 16S, HA1, HA3b and the mixture of HA1 and HA3 significantly increased interleukin 6 (IL6) production compared with NTX alone. Transcription of IL6 mRNA was low after stimulation with NTX alone, but increased to 16S-stimulation levels when NTX was mixed with HA1 or HA3b. In flow cytometry using labelled Abs against CD3 and CD19, the percentage of CD19 cells was higher following stimulation with 16S or NTX mixed with HA1 or HA3b compared with stimulation with NTX. The percentage of CD3 cells remained unchanged. These results suggest strongly that HA1 and HA3b demonstrate adjuvant activity via increasing IL6 production.