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Summary Preproteins are believed to be imported into chloroplasts through membrane contact sites where the translocon complexes of the outer (TOC) and inner (TIC) envelope membranes are assembled together. However, a single TOC–TIC supercomplex containing preproteins undergoing active import has not yet been directly observed. We optimized the blue native polyacrylamide gel electrophoresis (PAGE) (BN-PAGE) system to detect and resolve megadalton (MD)-sized complexes. Using this optimized system, the outer-membrane channel Toc75 from pea chloroplasts was found in at least two complexes: the 880-kD TOC complex and a previously undetected 1-MD complex. Two-dimensional BN-PAGE immunoblots further showed that Toc75, Toc159, Toc34, Tic20, Tic56 and Tic110 were all located in the 880-kD to 1.3-MD region. During active preprotein import, preproteins were transported mostly through the 1-MD complex and a smaller amount of preproteins was also detected in a complex of 1.25 MD. Antibody-shift assays showed that the 1-MD complex is a TOC–TIC supercomplex containing at least Toc75, Toc159, Toc34 and Tic110. Results from crosslinking and import with Arabidopsis chloroplasts suggest that the 1.25-MD complex is also a supercomplex. Our data provide direct evidence supporting that chloroplast preproteins are imported through TOC–TIC supercomplexes, and also provide the first size estimation of these supercomplexes. Furthermore, unlike in mitochondria where translocon supercomplexes are only transiently assembled during preprotein import, in chloroplasts at least some of the supercomplexes are preassembled stable structures.

Background Gold nanoparticles treated with near infrared (NIR) light can be heated preferentially, allowing for thermal ablation of targeted cells. The use of novel intravesical nanoparticle-directed therapy in conjunction with laser irradiation via a fiber optic cystoscope, represents a potential ablative treatment approach in patients with superficial bladder cancer. Objective To examine the thermal ablative effect of epidermal growth factor receptor (EGFR)-directed gold nanorods irradiated with NIR light in an orthotopic urinary bladder cancer model. Methods Gold nanorods linked to an anti-EGFR antibody (Conjugated gold NanoRods - CNR) were instilled into the bladder cavity of an orthotopic murine xenograft model with T24 bladder cancer cells expressing luciferase. NIR light was externally administered via an 808 nm diode laser. This treatment was repeated weekly for 4 weeks. The anti-cancer effect was monitored by an in vivo imaging system in a non-invasive manner, which was the primary outcome of our study. Results The optimal approach for an individual treatment was 2.1 W/cm2 laser power for 30 seconds. Using this in vivo model, NIR light combined with CNR demonstrated a statistically significant reduction in tumor-associated bioluminescent activity (n = 16) compared to mice treated with laser alone (n = 14) at the end of the study (p = 0.035). Furthermore, the CNR+NIR light treatment significantly abrogated bioluminescence signals over a 6-week observation period, compared to pre-treatment levels (p = 0.045). Conclusions Photothermal tumor ablation with EGFR-directed gold nanorods and NIR light proved effective and well tolerated in a murine in vivo model of urinary bladder cancer.

// Thomas W. Flaig 1 , Maren Salzmann-Sullivan 1 , Lih-Jen Su 1 , Zhiyong Zhang 1 , Molishree Joshi 2 , Miguel A. Gijon 2 , Jihye Kim 1 , John J. Arcaroli 1 , Adrie Van Bokhoven 3 , M. Scott Lucia 3 , Francisco G. La Rosa 3 and Isabel R. Schlaepfer 1 1 Division of Medical Oncology, Department of Medicine, University of Colorado, Anschutz Medical Campus, Aurora, Colorado, USA 2 Department of Pharmacology, University of Colorado, Anschutz Medical Campus, Aurora, Colorado, USA 3 Department of Pathology, University of Colorado, Anschutz Medical Campus, Aurora, Colorado, USA Correspondence to: Isabel R. Schlaepfer, email: // Keywords : CPT1A, prostate cancer, enzalutamide, ranolazine, INPP5K Received : February 27, 2017 Accepted : April 10, 2017 Published : April 21, 2017 Abstract Prostate cancer (PCa) is the most common malignancy among Western men and the second leading-cause of cancer related deaths. For men who develop metastatic castration resistant PCa (mCRPC), survival is limited, making the identification of novel therapies for mCRPC critical. We have found that deficient lipid oxidation via carnitine palmitoyltransferase (CPT1) results in decreased growth and invasion, underscoring the role of lipid oxidation to fuel PCa growth. Using immunohistochemistry we have found that the CPT1A isoform is abundant in PCa compared to benign tissue (n=39, p

Most chloroplast proteins are synthesized in the cytosol as higher molecular weight preproteins and imported via the translocons in the outer (TOC) and inner (TIC) envelope membranes of chloroplasts. Toc159 functions as a primary receptor and directly binds preproteins through its dimeric GTPase domain. As a first step toward a molecular understanding of how Toc159 mediates preprotein import, we mapped the preprotein-binding regions on the Toc159 GTPase domain (Toc159G) of pea (Pisum sativum) using cleavage by bound preproteins conjugated with the artificial protease FeBABE and cysteine-cysteine cross-linking. Our results show that residues at the dimer interface and the switch II region of Toc159G are in close proximity to preproteins. The mature portion of preproteins was observed preferentially at the dimer interface, whereas the transit peptide was found at both regions equally. Chloroplasts from transgenic plants expressing engineered Toc159 with a cysteine placed at the dimer interface showed increased cross-linking to bound preproteins. Our data suggest that, during preprotein import, the Toc159G dimer disengages and the dimer interface contacts translocating preproteins, which is consistent with a model in which conformational changes induced by dimer-monomer conversion in Toc159 play a direct role in facilitating preprotein import.

Visual interferences caused by bad weather conditions can have a negative impact on the performance of a visual surveillance system. How can we detect the moving object in a quickly and accurately manner is a very important step for further analysis, e.g., object recognition, object tracking, event detection, or behavior analysis, in a visual surveillance system. However most of the moving object object detection systems are based on a high-level micro-processor for its highly complex algorithm. Moreover, the detection accuracy is often affected by the environment of the system used. In this paper, we proposes using an FPGA (Field Programmable Gate Array) to realize a visual surveillance system so that the real-time requirement can be achieved. Besides, the proposed system can adapted itself to a variety of environmental situations, especially for the condition of light changes, such as the existence of a shadow, raindrop, etc. As we will see in the experiment that a very good performance in terms of power consumption, memory usage, throughput, as well as the capability of adapt to interference can be achieved in the proposed system

A person stays indoors for about 85%∼90% time of his lifetime, and the need for a comfortable indoor environment is getting higher; thus, the air-conditioning dependency becomes intense too. Nowadays, residents focus on both the comfortable living environment and indoor air quality. A closed environment will become hazardous because of carbon dioxide released during respiration and toxic organic solvent vapor released from interior decoration. In order to improve the indoor air quality (IAQ), we must allow outer fresh air into the indoor space and release the dirty air out. But while taking in fresh air, the heat and factory/vehicle exhaust are also introduced. Indoor CO2, HCHO, and VOCs and outer dirty gas threaten human health badly. To solve this problem, we bring up an innovative low-power-consuming full-outer-air-intake natural air-conditioning system that completely separates intake and exhaust air, which is a solution for cross-contamination and makes mass/energy exchange by means of air and water. Design airflow exceeds 300∼500 CFM, steam evaporation mass rate reaches 3.13∼3.88 kg/hr, and heat exchange capacity becomes 1,855∼2,300 kcal/hr. The sensible heat effectiveness is 71%∼112%, and EER exceeds 14.05∼17.42 kcal/W·h. In addition, the system under design can be of positive or negative pressure status according to the user’s or work’s requirement. It creates a comfortable and healthy living environment by supplying clean and fresh outer ambient air with low power consumption.

About 7% of people’s daily time is spent in taking vehicles between office and home. Besides, with the improvement of the living standard in today’s society, people’s requirements for a comfortable environment inside the car are constantly increasing and this must rely on an effective vehicle air conditioner to maintain the comfort of the cabin environment. In general, a vehicle air conditioner uses the air-mixing mode to regulate the temperature control system. In this mode of operation, the compressor needs to work continuously, which is extremely energy consuming. The vehicle’s air conditioner is greatly affected by the inner and outer heat load, which are generated therein. Furthermore, the heat load is instantly changeable. Therefore, only when the controller can adapt to the feature of heat load, then we can find the optimal control method, thus enabling the vehicle’s air conditioner to interact with the actual heat load to supply the balanced cooling capacity and, as a result, create the most comfortable environment inside the cabin with minimum energy consumption. For this purpose, we bring up in this paper a low-energy-consumption smart vehicle air-conditioning control system to detect total heat load, which can change the vehicle’s air-conditioning capacity mode to maintain the average temperature at 25.2°C∼26.2°C and the average humidity at 46.6%∼54.4% in the cabin. When the inner heat load is stable, the rest times of the compressor can reach 16∼23 times per hour, which attains a rate of fuel saving around 21%∼28%. With the proposed architecture, the purpose of the low-energy-consumption vehicle air-conditioning system can be achieved, which, at the same time, creates a comfortable environment inside the cabin.

This paper reports successful synthesis of multifunctional nanoclusters of upconversion nanoparticle (UCNP) and gold nanorod (AuNR) through a PEGylation process. UCNPs emit visible luminescence under near-infrared excitation, producing high-contrast images with no background fluorescence. When coupled with AuNRs, the resulting UCNP-AuNR multifunctional nanoclusters are capable of simultaneous detection and treatment of bladder cancer. These UCNP-AuNR nanoclusters are further functionalized with antibodies to epidermal growth factor receptor (EGFR) to target bladder cancer cells known to overexpress EGFRs. This paper demonstrates, for the first time, efficient targeting of bladder cancer cells with UCNP-AuNR nanoclusters. In addition to high-contrast imaging and consequently high sensitivity detection of bladder cancer cells, highly selective optoporation-assisted chemotherapy was accomplished using a dosage of chemotherapy agent significantly lower than any previous reports, within a clinically relevant incubation time window. These results are highly relevant to the eventual human application in which the nanoclusters and chemotherapy drugs will be directly instilled in bladder via urinary catheter.

The purpose of this study was to gather the perceptions and the attitudes of elementary school teachers in Taiwan R.O.C. regarding inservice computer education programs. The study also investigated elementary school teachers' attitudes towards content, format, scheduling, and administrative' support of inservice education. Data were collected by way of a survey which was measured using a Likert scale. A proportional, stratified random sampling procedure was used to identify 200 elementary schools from the 2,467 elementary schools in Taiwan. The survey was mailed to principals of sampled schools. Only one teacher from each school was chosen as a subject. A response rate of 84% was achieved. Eleven hypotheses were tested at the 0.05 level of significance by means of one-way ANOVAs, chi-squares, t-tests, and Friedman rank sum tests. Major findings were that (1) elementary school teachers should receive inservice credits from colleges or universities for updating professional knowledge to improve classroom performance; (2) teachers need to be involved in the development of purposes, activities, and evaluation methods; (3) educational agencies should provide sufficient inservice programs for elementary school teachers; (4) private elementary school teachers express a stronger need for opportunities to select appropriate inservice programs and for cooperation from administrators, parents of students, and staff than did public elementary school teachers; (5) teachers with more experience in inservice computer education expect more advanced and challenging courses; and (6) teachers' colleges play an important role in providing appropriate inservice computer education programs for elementary school teachers. It was recommended that (a) inservice computer education provide courses of different levels, depending upon the computer knowledge of those enrolled; (b) inservice computer education for elementary school teachers focus on teacher utilities, application packages, computer assisted instruction, computer educational applications, computer operation, and structure; (c) administrators or principals have the first opportunities to participate in inservice computer education; and (d) a nation-wide research center be established to conduct educational studies and necessary follow-up research.

The two-membrane envelope is a defining feature of chloroplasts. Chloroplasts evolved from a Gram-negative cyanobacterial endosymbiont. During evolution, genes of the endosymbiont have been transferred to the host nuclear genome. Most chloroplast proteins are synthesized in the cytosol as higher-molecular-mass preproteins with an N-terminal transit peptide. Preproteins are transported into chloroplasts by the TOC and TIC (translocons at the outer- and inner-envelope membranes of chloroplasts, respectively) machineries1,2, but how TOC and TIC are assembled together is unknown. Here we report the identification of the TIC component TIC236; TIC236 is an integral inner-membrane protein that projects a 230-kDa domain into the intermembrane space, which binds directly to the outer-membrane channel TOC75. The knockout mutation of TIC236 is embryonically lethal. In TIC236-knockdown mutants, a smaller amount of the inner-membrane channel TIC20 was associated with TOC75; the amount of TOC-TIC supercomplexes was also reduced. This resulted in a reduced import rate into the stroma, though outer-membrane protein insertion was unaffected. The size and the essential nature of TIC236 indicate that-unlike in mitochondria, in which the outer- and inner-membrane translocons exist as separate complexes and a supercomplex is only transiently assembled during preprotein translocation3,4-a long and stable protein bridge in the intermembrane space is required for protein translocation into chloroplasts. Furthermore, TIC236 and TOC75 are homologues of bacterial inner-membrane TamB5 and outer-membrane BamA, respectively. Our evolutionary analyses show that, similar to TOC75, TIC236 is preserved only in plants and has co-evolved with TOC75 throughout the plant lineage. This suggests that the backbone of the chloroplast protein-import machinery evolved from the bacterial TamB-BamA protein-secretion system.

Upconverting nanophosphors (UCNPs) absorb two or more photons at 980nm and emit a higher energy photon in the visible range. UCNPs provide distinct advantages as biological imaging agents in that they have no autofluorescence, don’t photobleach, are capable of deep tissue imaging, exhibit no blinking, and are physically robust. Through surface modifications with an amphiphilic polymer, we can not only functionalize UCNPs for targeted imaging, but also tether them to plasmonic nanoparticles such as gold nanorods (AuNRs) for enhanced upconversion and multifunctionality. We developed a process to construct UCNP-AuNR nanoclusters using polyethylene glycol. The UCNP-AuNR nanocluster is further modified with an anti-epidermal growth factor receptor (aEGFR), allowing specific binding to bladder cancer cells that highly express epidermal growth factor receptor. Once the nanoclusters bind to the cell membrane we can (1) perform targeted and high contrast imaging of the bladder cancer cells and (2) utilize localized surface plasmon of AuNRs to selectively kill the cells in situ upon detection by UCNP fluorescence. Successful conjugation and integrity of the UCNP-AuNR nanoclusters were confirmed via electron microscopy. Then, through a combination of brighftield, confocal upconversion fluorescence, and infrared darkfield microscopy we demonstrate selective binding and high-contrast upconversion imaging of the bladder cancer cells. Finally, through a series of in vitro studies, we demonstrate two different methods of cell killing. First, with a continuous wave laser, we demonstrate effective thermal ablation of cells. Second, with a femtosecond pulse laser, we demonstrate optoporation of the cell membrane that allows increased uptake of drugs.

Three stromal chaperone ATPases, cpHsc70, Hsp90C, and Hsp93, are present in the chloroplast translocon, but none has been shown to directly bind preproteins in vivo during import, so it remains unclear whether any function as a preprotein-translocating motor and whether they have different functions during the import process. Here, using protein crosslinking followed by ionic detergent solubilization, we show that Hsp93 directly binds to the transit peptides of various preproteins undergoing active import into chloroplasts. Hsp93 also binds to the mature region of a preprotein. A time course study of import, followed by coimmunoprecipitation experiments, confirmed that Hsp93 is present in the same complexes as preproteins at an early stage when preproteins are being processed to the mature size. In contrast, cpHsc70 is present in the same complexes as preproteins at both the early stage and a later stage after the transit peptide has been removed, suggesting that cpHsc70, but not Hsp93, is important in translocating processed mature proteins across the envelope.

This paper proposes body posture recognition and turning recording system for assisting the care of bed bound patients in nursing homes. The system continuously detects the patient's body posture and records the length of time for each body posture. If the patient remains in the same body posture long enough to develop pressure ulcers, the system notifies caregivers to change the patient's body posture. The objective of recording is to provide the log of body turning for querying of patients' family members. In order to accurately detect patient's body posture, we developed a novel pressure sensing pad which contains force sensing resistor sensors. Based on the proposed pressure sensing pad, we developed a bed posture recognition module which includes a bed posture recognition algorithm. The algorithm is based on fuzzy theory. The body posture recognition algorithm can detect the patient's bed posture whether it is right lateral decubitus, left lateral decubitus, or supine. The detected information of patient's body posture can be then transmitted to the server of healthcare center by the communication module to perform the functions of recording and notification. Experimental results showed that the average posture recognition accuracy for our proposed module is 92%.

Introduction The optimal dose of inhaled metered-dose bronchodilators for intubated patients with chronic obstructive pulmonary disease (COPD) is unknown. In this study, we proposed a bronchodilator dosing schedule based on an individual's airway resistance (Raw) and tested its efficacy in reducing Raw. Methods A total of 51 newly admitted patients with invasively ventilated COPD were randomly assigned to receive personalized or fixed bronchodilator dosing. Personal target Raw was defined by measuring each individual's Raw after maximal pharmacologic bronchodilatation. Thereafter, Raw was measured every 8 h until the 28th day. Patients in the fixed-dosing group received only predetermined doses. Additional doses of bronchodilators were given to patients in the personalized-dosing group when the measured Raw exceeded their target Raw. Results The median daily doses of salmeterol/fluticasone were 9.2 (personalized-dosing) vs 7.6 (fixed-dosing) puffs (P Conclusion Personalized dosing of inhaled bronchodilator administered to invasively ventilated COPD patients can produce a better reduction in Raw. Further studies with larger sample size are required to verify the conclusion of this pilot study.

This paper presents a robotic shopping cart for a shopping mall to provide shopping assistance and information. When customers login to the shopping cart system, the information providing is first started to provide directions and recommendations of the shopping mall. In addition, the robotic shopping cart is developed to be autonomous returning to starting position while customers logout from the service screen. Finally, the purchase record of customers can be send to the centralized system of shopping mall for further analysis of various shopping behaviors.

In this paper we propose a scalable face image compression algorithm based on Principal Component Analysis (PCA) and Entropy Coding. By using PCA and some training face image patterns, we can extract the most representative eigen-image of human faces. To reduce the coding complexity as well as to achieve a higher compression ratio, only the first term of the extracted eigen-images will be used for the encoding of the human face, i.e., only the eigen-image with maximal energy strength will be selected for the encoding process. As we will see in the experiment that a good trade off between the computation complexity, compression ratio, and image quality can be achieved with the proposed algorithm.

Generally, different students employ different learning styles dur-ing their studies and medical students are exposed to diverse methods of teaching. Therefore, understanding students’ learning style preference is an important consideration for a high quality and effective teaching and learning process.The aim of the study was to study the variation of learning styles among pre-clinical medical students of SEGi University, Malaysia. A cross-sectional study was performed by using VARK (Visual, Audio, Reading and Kinaesthetic) questionnaire version 7.2 to assess the learning style preference of 98 (n=98) pre-clinical medical students in SEGi University. The questionnaire consists of 16 items which identify four different learning styles: visual, aural, reading/writing and kin-esthetic. Descriptive statistics were used to identify the learning styles of students. 61 students preferred multimodal as their learning style, out of which 43 (70%) of them were female stu-dents and 18 (30%) were male students. 37 students preferred unimodal as their learning style out of which 22 (59%) of them were female students and 15 (41%) were male students. In addi-tion, female students had more diverse preferences than male students by having 10 out of the other 11 possible combinations in multimodal learning style of preference, whereas the male stu-dents only had 5 out of the 11 combinations. In this study, there was no significant gender difference in the percentages of males and female students who preferred unimodal and multimodal styles of information presentation (P= 0.263; α=0.05). To con-clude, the majority of students of both genders had chosen quad-modal as their learning style preference. The results of this study can provide useful information for improving the quality of the teaching and learning experiences of students.

Tic110 is a major component of the chloroplast protein import translocon. Two functions with mutually exclusive structures have been proposed for Tic110: a protein-conducting channel with six transmembrane domains and a scaffold with two N-terminal transmembrane domains followed by a large soluble domain for binding transit peptides and other stromal translocon components. To investigate the structure of Tic110, Tic110 from Cyanidioschyzon merolae (CmTic110) was characterized. We constructed three fragments, CmTic110A , CmTic110B and CmTic110C , with increasing N-terminal truncations, to perform small-angle X-ray scattering (SAXS) and X-ray crystallography analyses and Dali structural comparison. Here we report the molecular envelope of CmTic110B and CmTic110C determined by SAXS, and the crystal structure of CmTic110C at 4.2 Å. Our data indicate that the C-terminal half of CmTic110 possesses a rod-shaped helix-repeat structure that is too flattened and elongated to be a channel. The structure is most similar to the HEAT-repeat motif that functions as scaffolds for protein-protein interactions.

An efficient approach to the sharpening of color images is proposed in this paper. For this, the image to be sharpened is first transformed to theHSVcolor model, and then only the channel ofValuewill be used for the process of sharpening while the other channels are left unchanged. We then apply a proposed edge detector and low-pass filter to the channel ofValueto pick out pixels around boundaries. After that, those pixels detected as around edges or boundaries are adjusted so that the boundary can be sharpened, and those nonedge pixels are kept unaltered. The increment or decrement magnitude that is to be added to those edge pixels is determined in an adaptive manner based on global statistics of the image and local statistics of the pixel to be sharpened. With the proposed approach, the discontinuities can be highlighted while most of the original information contained in the image can be retained. Finally, the adjusted channel ofValueand that ofHueandSaturationwill be integrated to get the sharpened color image. Extensive experiments on natural images will be given in this paper to highlight the effectiveness and efficiency of the proposed approach.

We construct a robust and precise multi-orientation text detection system in scene images which can extensively locate possible characters with multi-information fusion. In our method, an adaptive multi-channel character grouping algorithm is first proposed to extract all possible character candidates robustly, and an AdaBoost classifier is then to properly identify character candidates as characters or non-characters. A single-link clustering with distance metric learning is thereafter used to adaptively group characters into text regions, and an effective hybrid filter with Convolution Neural Networks (CNN), AdaBoost and Bayesian classifiers is finally designed to precisely verify the extracted text regions. Our proposed technology is extensively evaluated on several public multi-orientation scene text datasets, e.g., MSRA-TD500 and USTB-SV1K, and is much better than state-of-the-art methods.

NaYF4:Yb3+,Er3+ upconverting nanophosphors (UCNPs) are robust and stable nanoparticles that absorb near-infrared (NIR) photons and emit green and red visible photons through energy transfer upconversion. This mechanism provides UCNPs several advantages as a bioimaging agent over traditional fluorescence imaging agent in that NIR excitation allows high-contrast imaging without autofluorescence and that they can be used for deep-tissue imaging. However, additional surface modification of UCNPs is necessary for them to be biocompatible. We use an amphiphilic polymer (poly(maleic anhydride-alt-octadecene) (PMAO) and a hetero-functional polyethylene glycol with amine and thiol ends (NH2-PEG-SH)) to make the UCNPs water-soluble. This reaction yields a carboxylic group that allows functionalization with anti-epidermal growth factor receptor (aEGFR), which provides specific binding of UCNPs to EGFR-expressing bladder cancer cells. Additionally, the thiol ends of the PEGylated UCNPs are able to bind with gold nanorods (AuNRs) to create UCNP-AuNR complexes. The localized surface plasmon of the AuNR then allow localized heating of HTB9 bladder cancer cells, enabling in situ cell killing upon detection by UCNP fluorescence. Here, we report a successful synthesis, surface modification and conjugation of aEGFR functionalized UCNP-AuNR complexes and in vitro imaging and thermal ablation studies using them. Synthesis and surface modification of UCNP-AuNR complexes are confirmed by electron microscopy. Then, a combination of brightfield, NIR confocal fluorescence, and darkfield microscopy on the UCNP-AuNR treated bladder cancer cells revealed successful cancer targeting and imaging capabilities of the complex. Finally, cell viability assay showed that NIR irradiation of UCNP-AuNR conjugated cells resulted highly selective cell killing.

Toc75 is the channel for protein translocation across the chloroplast outer envelope membrane. Toc75 belongs to the Omp85 protein family and consists of three N-terminal polypeptide transport-associated (POTRA) domains that are essential for the functions of Toc75, followed by a membrane-spanning β-barrel domain. In bacteria, POTRA domains of Omp85 family members are located in the periplasm, where they interact with other partner proteins to accomplish protein secretion and outer membrane protein assembly. However, the orientation and therefore the molecular function of chloroplast Toc75 POTRA domains remain a matter of debate. We investigated the topology of Toc75 using bimolecular fluorescence complementation and immunogold electron microscopy. Bimolecular fluorescence complementation analyses showed that in stably transformed plants, Toc75 N terminus is located on the intermembrane space side, not the cytosolic side, of the outer membrane. Immunogold labeling of endogenous Toc75 POTRA domains in pea (Pisum sativum) and Arabidopsis (Arabidopsis thaliana) confirmed that POTRA domains are located in the intermembrane space of the chloroplast envelope.

Recognition of micro-RNA function and their contribution to the biology of disease has given a new insight into disease mechanisms, with these discoveries potentially improving clinical diagnostic and therapeutic options. miR-125b has been identified as an important regulator in various cancers, including prostate cancer, but the mechanism of this regulation remains incompletely understood. In these studies, the effect of castration on miR-125b serum expression was evaluated in mice, simulating androgen deprivation. Furthermore, miR-125b expression was measured by quantitative real-time polymerase chain reaction (qRT-PCR) in LNCaP prostate cancer cells treated with the antiandrogen bicalutamide. Using LNCaP cells, the effect of miR-125b modulation on apoptotic protein and NCOR2, a co-repressor of androgen receptor (AR), was examined by Western blot. A 3′-untranslated region (UTR) luciferase-binding assay was performed to confirm that miR-125b targets NCOR2. We found that surgical castration induced an initial increase in the expression of circulating miR-125b in mice, while sham surgery did not. In addition, AR blockade via bicalutamide was associated with the rapid release of miR-125b into the cell culture medium of prostate cancer cells. A previously studied target of miR-125b, a regulator in the apoptotic pathway, BAK1, could not completely account for the role of miR-125b in prostate cancer. Thus, we looked for additional targets of miR-125b and found that NCOR2, which is a repressor of AR, is a direct target of miR-125b. We found that NCOR2 protein expression was blocked by mimics of miR-125b, and a luciferase-binding assay confirmed that NCOR2 is a direct target of miR-125b. Our data provide novel evidence that miR-125b is an important regulator of the AR with specific ramification for the effectiveness of antiandrogens and other hormonal therapies in prostate cancer.