Miracle-dz/UniFAR-FTUs huggingface dataset:
Open Conformation of Ezrin Bound to Phosphatidylinositol 4,5-Bisphosphate and to F-actin Revealed by Neutron Scattering*

Background: The structure of activated ezrin is not known. Results: We have determined the conformation of activated ezrin upon binding to PIP2 and to F-actin. Conclusion: Activated ezrin forms more extensive contacts with F-actin than generally depicted. Significance: This study provides new insight into the mechanisms by which ezrin assembles signaling complexes at the membrane-cytoskeleton interface. Ezrin is a member of the ezrin-radixin-moesin family (ERM) of adapter proteins that are localized at the interface between the cell membrane and the cortical actin cytoskeleton, and they regulate a variety of cellular functions. The structure representing a dormant and closed conformation of an ERM protein has previously been determined by x-ray crystallography. Here, using contrast variation small angle neutron scattering, we reveal the structural changes of the full-length ezrin upon binding to the signaling lipid phosphatidylinositol 4,5-bisphosphate (PIP2) and to F-actin. Ezrin binding to F-actin requires the simultaneous binding of ezrin to PIP2. Once bound to F-actin, the opened ezrin forms more extensive contacts with F-actin than generally depicted, suggesting a possible role of ezrin in regulating the interfacial structure and dynamics between the cell membrane and the underlying actin cytoskeleton. In addition, using gel filtration, we find that the conformational opening of ezrin in response to PIP2 binding is cooperative, but the cooperativity is disrupted by a phospho-mimic mutation S249D in the 4.1-ezrin/radixin/moesin (FERM) domain of ezrin. Using surface plasmon resonance, we show that the S249D mutation weakens the binding affinity and changes the kinetics of 4.1-ERM to PIP2 binding. The study provides the first structural view of the activated ezrin bound to PIP2 and to F-actin.
Relationship and associated mechanisms between ambulatory blood pressure and clinic blood pressure with prevalent cardiovascular disease in diabetic hypertensive patients

Abstract The present study was to compare the association between ambulatory blood pressure (ABP) and clinic BP (CBP) with prevalent cardiovascular diseases (CVD); and the underlying mechanism would also be investigated concurrently. Diabetic hypertensive patients were enrolled and divided into 2 groups based on presence of CVD. Twenty-four hour-ABP monitoring was performed and between-group differences were evaluated and logistic regression analysis was conducted. A total of 568 diabetic hypertensive patients were enrolled, and the mean age was 60.8 years, male accounted for 67.8%. Mean durations of diabetes mellitus and hypertension were 6.1 ± 2.7 and 5.4 ± 3.3 years, respectively, and 20.6% had prevalent CVD. Compared to patients without CVD, patients with CVD had significantly higher body mass index (BMI), plasma aldosterone concentration (PAC), and serum sodium level. No significant between-group differences in CBP were observed. However, 24 hour-SBP, daytime-SBP and nighttime-SBP were all significantly higher in patients with CVD compared to those without CVD. Pearson correlation analysis showed that BMI was positively correlated with PAC and serum sodium level. Logistic regression analyses showed that the association between clinic SBP and DBP with CVD were progressively attenuated to nonsignificant. In contrast, both ambulatory SBP and DBP were independently associated with CVD. However, after being further adjusted for PAC, no significant association was observed between ambulatory SBP and CVD. In diabetic hypertensive patients, ABP is superior to CBP in relation to CVD. The association between ambulatory SBP and CVD may be dependent on aldosterone excess.
Solution structure of Escherichia coli glutaredoxin-2 shows similarity to mammalian glutathione-S-transferases.

Glutaredoxin 2 (Grx2) from Escherichia coli is distinguished from other glutaredoxins by its larger size, low overall sequence identity and lack of electron donor activity with ribonucleotide reductase. However, catalysis of glutathione (GSH)-dependent general disulfide reduction by Grx2 is extremely efficient. The high-resolution solution structure of E. coli Grx2 shows a two-domain protein, with residues 1 to 72 forming a classical "thioredoxin-fold" glutaredoxin domain, connected by an 11 residue linker to the highly helical C-terminal domain, residues 84 to 215. The active site, Cys9-Pro10-Tyr11-Cys12, is buried in the interface between the two domains, but Cys9 is solvent-accessible, consistent with its role in catalysis. The structures reveal the hither to unknown fact that Grx2 is structurally similar to glutathione-S-transferases (GST), although there is no obvious sequence homology. The similarity of these structures gives important insights into the functional significance of a new class of mammalian GST-like proteins, the single-cysteine omega class, which have glutaredoxin oxidoreductase activity rather than GSH-S-transferase conjugating activity. E. coli Grx 2 is structurally and functionally a member of this new expanding family of large glutaredoxins. The primary function of Grx2 as a GST-like glutaredoxin is to catalyze reversible glutathionylation of proteins with GSH in cellular redox regulation including stress responses.
Demonstration by a Novel Genetic Technique That Leader Peptidase Is an Essential Enzyme of Escherichia coli

It was previously shown that two separate regions of DNA are required for expression of the cloned leader peptidase gene on plasmid pTD101 (T. Date and W. Wickner, Proc. Natl. Acad. Sci. U.S.A. 78:6106-6110, 1981). Both loci have been mapped in detail, and their roles have been established. A 1.3-kilobase region, termed the L region, encodes the 37,000-dalton leader peptidase protein. Another region, termed the P region, is about 1.5 kilobases away from the L region and is less than 350 base pairs long. The P region acts in cis to the L region, suggesting that it plays a role as a promoter. A technique for inactivation of the leader peptidase gene on the Escherichia coli chromosome has been developed to examine whether the leader peptidase which we had cloned is essential for cell growth. A specific plasmid (P− L−) which deletes both the P region and a substantial portion of the L region was constructed and transformed into a polA mutant strain. The plasmid cannot replicate in this strain; thus, the plasmid-borne ampicillin resistance is lost unless the plasmid DNA recombines into the chromosome. Integration of the P− L− plasmid did not yield any viable ampicillin-resistant cells, whereas the three control plasmids, P+ L+, P+ L−, and P− L+, did. When the P− L− plasmid was transformed into the polA(Ts) strain, the strain could only grow in the presence of ampicillin at a permissive temperature, suggesting that integration of the plasmid into the host chromosome leads to inactivation of the chromosomal leader peptidase gene. Southern hybridization analysis demonstrated that the integration of plasmids into the chromosome occurred at the homologous site. This study demonstrates that expression of the leader peptidase gene is critical for cell growth.
Proteasomal degradation of myocardin is required for its transcriptional activity in vascular smooth muscle cells

Myocardin is a transcriptional co‐activator of serum response factor (SRF) and can be degraded through ubiquitin–proteasome system. Our preliminary studies unexpectedly revealed that accumulation of myocardin in response to proteasome inhibition by MG132 or lactacystin resulted in decrease of transcriptional activity of myocardin as indicated by reduced expression of SMC contractile marker genes (SM α‐actin, SM22, and calponin) and muscle‐enriched microRNAs (miR‐143/145 and miR‐1/133a), and reduced contractility of human vascular smooth muscle cells (SMCs) embedded in collagen gel lattices, suggesting that myocardin degradation is required for its transcriptional activity. Further studies using chromatin immunoprecipitation assay revealed that proteasome inhibition, although increased the occupancy of myocardin and SRF on the promoter of SM α‐actin gene, abolished myocardin‐dependent recruitment of RNA polymerase II. We further examined the degradation of myocardin in epithelioid and spindle‐shaped SMCs and revealed that myocardin in more differentiated spindle‐shaped SMCs was more quickly degraded and had shorter half‐life than in epithelioid SMCs. In neointimal lesions, we found that stabilization of myocardin protein was companied by downregulation of transcripts of ubiquitin and proteasome subunits, further illustrating the mechanism underlying reduction of myocardin transcriptional activity. In summary, our results have suggested that proteasomal degradation of myocardin is required for its transcriptional activity. J. Cell. Physiol. 226: 1897–1906, 2011. © 2010 Wiley‐Liss, Inc.
Effect of yoga therapy on facial emotion recognition deficits, symptoms and functioning in patients with schizophrenia

Behere RV, Arasappa R, Jagannathan A, Varambally S, Venkatasubramanian G, Thirthalli J, Subbakrishna DK, Nagendra HR, Gangadhar BN. Effect of yoga therapy on facial emotion recognition deficits, symptoms and functioning in patients with schizophrenia.
Inequalities in Indigenous Oral Health: Findings from Australia, New Zealand, and Canada

The objective was to compare absolute differences in the prevalence of Indigenous-related inequalities in dental disease experience and self-rated oral health in Australia, Canada, and New Zealand. Data were sourced from national oral health surveys in Australia (2004 to 2006), Canada (2007 to 2009), and New Zealand (2009). Participants were aged ≥18 y. The authors measured age- and sex-adjusted inequalities by estimating absolute prevalence differences and their corresponding 95% confidence intervals (95% CIs). Clinical measures included the prevalence of untreated decayed teeth, missing teeth, and filled teeth; self-reported measures included the prevalence of “fair” or “poor” self-rated oral health. The overall pattern of Indigenous disadvantage was similar across all countries. The summary estimates for the adjusted prevalence differences were as follows: 16.5 (95% CI: 11.1 to 21.9) for decayed teeth (all countries combined), 18.2 (95% CI: 12.5 to 24.0) for missing teeth, 0.8 (95% CI: –1.9 to 3.5) for filled teeth, and 17.5 (95% CI: 11.3 to 23.6) for fair/poor self-rated oral health. The I2 estimates were small for each outcome: 0.0% for decayed, missing, and filled teeth and 11.6% for fair/poor self-rated oral health. Irrespective of country, when compared with their non-Indigenous counterparts, Indigenous persons had more untreated dental caries and missing teeth, fewer teeth that had been restored (with the exception of Canada), and a higher proportion reporting fair/poor self-rated oral health. There were no discernible differences among the 3 countries.
Widespread Distribution of a Tet W Determinant among Tetracycline-Resistant Isolates of the Animal Pathogen Arcanobacterium pyogenes

ABSTRACT Tetracycline resistance is common among isolates of the animal commensal and opportunistic pathogen Arcanobacterium pyogenes. The tetracycline resistance determinant cloned from two bovine isolates of A. pyogenes was highly similar at the DNA level (92% identity) to the tet(W) gene, encoding a ribosomal protection tetracycline resistance protein, from the rumen bacterium Butyrivibrio fibrisolvens. The tet(W) gene was found in all 20 tetracycline-resistant isolates tested, indicating that it is a widely distributed determinant of tetracycline resistance in this organism. In 25% of tetracycline-resistant isolates, the tet(W) gene was associated with a mob gene, encoding a functional mobilization protein, and an origin of transfer, suggesting that the determinant may be transferable to other bacteria. In fact, low-frequency transfer of tet(W) was detected from mob+ A. pyogenes isolates to a tetracycline-sensitive A. pyogenes recipient. The mobile nature of this determinant and the presence of A. pyogenes in the gastrointestinal tract of cattle and pigs suggest that A. pyogenes may have inherited this determinant within the gastrointestinal tracts of these animals.
Development and Validation of a Simple Isocratic HPLC Method for Simultaneous Estimation of Phytosterols in Cissus quadrangularis

Cissus quadrangularis L. is a promising remedy prescribed in the ancient Ayurvedic literature for bone fracture healing properties. As this activity has been extensively investigated and well established, a range of formulations containing C. quadrangularis has been marketed. This work reports the development and validation of a reliable RP-HPLC method for the analysis of phytosterols in the various extracts of the plant. The proposed method utilizes a Cosmosil C8 column (250 ΄ 4.6 mm) with a compatible Phenomenex C8 guard column with isocratic elution of acetonitrile and water (95:5 v/v) at 25°. An effluent flow rate of 2 ml/min and UV detection at 202 nm was used for the analysis of phytosterols. The described method was linear in the range of 1–500 μg/ml, with excellent correlation coefficients. The precision, robustness and ruggedness values were also within the prescribed limits (less than 2%). The recovery values were within the range, which indicates that the accuracy of the analysis was good and that the interference of the matrix with the recovery of phytosterols was low. The phytosterols were found to be stable in a stock solution for 48 h (% RSD was below 2%) and no interfering extra peaks were observed under controlled stress conditions. The proposed method is simple, specific, precise, accurate, and reproducible and thus can be used for routine analysis of C. quadrangularis phytosterols in quality control laboratories.
DOES SIZE MATTER MOST? THE EFFECT OF GROWTH HISTORY ON PROBABILISTIC REACTION NORM FOR SALMON MATURATION

Abstract Body size is widely believed to affect the occurrence of sexual maturation. Recent studies have used changes in the age-specific body size at which the probability of maturing is 50%, a feature of probabilistic reaction norms, to quantify purported evolution of life histories. However, body size results from a combination of growth rates during successive developmental stages. Therefore, to understand the evolution of the maturation schedule, it is necessary to comprehend the relationships among body size, growth history, and maturation schedule. We examined the relationships among body size, previous growth history, and maturation probability in chum salmon (Oncorhynchus keta). In this study, previous growth history was estimated from yearly specific growth increments that provide information describing body size. Previous growth history was found to be more closely linked to maturation probability than body size. The most recent growth condition was the most important factor affecting whether a fish matured during the subsequent breeding season. Because individuals of similar body size and same age can have different growth histories, the relationship between body size and maturation probability could be plastically modified by growth history. This may violate an assumption required to infer evolution, namely that size-related maturation trends in probabilistic reaction norms are immune to growth history.
Thioredoxin-Interacting Protein Mediates Apoptosis in Early Brain Injury after Subarachnoid Haemorrhage

Early brain injury (EBI) is considered to be the major factor associated with high morbidity and mortality after subarachnoid haemorrhage (SAH). Apoptosis is the major pathological mechanism of EBI, and its pathogenesis has not been fully clarified. Here, we report that thioredoxin-interacting protein (TXNIP), which is induced by protein kinase RNA-like endoplasmic reticulum (ER) kinase (PERK), participates in EBI by promoting apoptosis. By using adult male Sprague-Dawley rats to establish SAH models, as well as Terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) staining, immunofluorescence, and western blot, we found that TXNIP expression significantly increased after SAH in comparison to the sham group and peaked at 48 h (up to 3.2-fold). Meanwhile, TXNIP was widely expressed in neurons and colocalized with TUNEL-positive cells in the hippocampus and cortex of SAH rats. After administration of TXNIP inhibitor-resveratrol (60 mg/kg), TXNIP small interfering RNA (siRNA) and the PERK inhibitor GSK2656157, TXNIP expression was significantly reduced, accompanied by an attenuation of apoptosis and prognostic indicators, including SAH grade, neurological deficits, brain water content, and blood-brain barrier (BBB) permeability. Collectively, these results suggest that TXNIP may participate in EBI after SAH by mediating apoptosis. The blockage of TXNIP induced by PERK could be a potential therapeutic strategy for SAH treatment.
Solution structure of the fifth repeat of factor H: a second example of the complement control protein module.

Modules which share the same consensus sequence are assumed to have common structural features, at the secondary and tertiary level. In order to test the extent of such similarities, it is necessary to examine the structures of several examples from each module family. Recently, the first three-dimensional structure of a complement control protein (CCP) module (the 16th repeat of human factor H, H16) was determined using a combination of two-dimensional NMR and simulated annealing [Norman, D.G. , Barlow, P.N. , Baron, M. , Day, A.J., Sim, R.B. , & Campbell, I.D. (1991) J. Mol. Biol. 219, 717-725]. Using the same techniques, the three-dimensional structure of a second CCP module (the 5th repeat of human factor H, H5) has now been determined. The primary sequence of H5 contains 17 residues which are identical and in equivalent position to those in H16. Thirteen of these 17 are part of the consensus sequence. The similarities between the secondary structure of H5 and that of H16 are extensive. This implies that the consensus sequence dictates a particular secondary structure. The tertiary structure of H5, a compact hydrophobic core wrapped in beta-strand and sheet, bears much overall resemblance to that of H16. However, there is a deletion in the first strand of H5, and an insertion in a loop, resulting in slightly shorter overall length. This is associated with a rearrangement of residues within the hydrophobic core. The side chain of the highly conserved Tyr29, which occupies a central position within the core of H16, lies on the periphery of the core of H5.
Effects of Lower Extremity Passive Stretching: Pilot Study of Children and Youth with Severe Limitations in Self-Mobility

Purpose The purpose of this pilot study was to investigate changes in passive range of motion (PROM) of children with severe limitations in self-mobility during periods of physical therapy intervention and naturally occurring periods of no intervention (school vacations). Methods A multiple-case time-series design was used to monitor hip and knee PROM over a seven-month period for seven students aged four to 18 years receiving physical therapy in a school setting. PROM measurements were graphed and the Wilcoxon signed rank test was used to evaluate the PROM changes during intervention and nonintervention phases for each individual student and for the students as a group. Results When PROM changes were pooled for all subjects and all joint motions, the Wilcoxon signed rank test revealed a significant decrease in PROM only after the first nonintervention phase. Significant decreases in PROM were observed in only two of the students after the first nonintervention phase. Conclusion Children and youth with severe limitations in self-mobility may lose PROM during nonintervention periods lasting more than five weeks.
Posaconazole Treatment in Hematology Patients: A Pilot Study of Therapeutic Drug Monitoring

Background: Posaconazole is indicated for prophylaxis and salvage therapy of invasive fungal infections. Based on pharmacokinetic–pharmacodynamic data, minimum serum concentrations for each indication have been proposed, for example, for prophylaxis >0.5–0.7 mg/L and for primary therapy >1.0 mg/L. Several drugs and comorbidities have been identified to hinder reaching target concentrations. It is postulated that patients with interacting drugs or comorbidities should be monitored for posaconazole concentrations. Patients and Methods: Patients aged 18 years and above were included for retrospective analysis if at least 1 serum posaconazole concentration was measured in our hospital between June 2009 and May 2010. Serum posaconazole concentrations were measured using a validated liquid chromatographic method with tandem mass-spectrometric analytical method. Patient characteristics, underlying disease, comedication, comorbidities, and therapeutic drug monitoring (TDM) interventions were collected retrospectively, based on (electronic) medical records. Results: Seventeen patients were included, from whom 42 samples for posaconazole measurement were collected. In total, 8 patients did not reach adequate posaconazole concentration. Sixty percent of patients using a proton pump inhibitor (PPI) did not reach target concentration with a corresponding median concentration of 0.48 mg/L. PPI usage was shown to significantly increase the risk of attaining below-target serum posaconazole concentration (P = 0.04 for all measurements). Graft-versus-host disease and diarrhea were associated with significant below-target concentrations (P = 0.03 and P < 0.001, respectively, for all measurements). One patient developed a breakthrough pulmonary aspergillosis at low posaconazole concentration (0.37 mg/L). Two patients had high concentrations (>3 mg/L), without adverse events. After TDM intervention, 3 out of 4 patients (75%) reached target concentration by spreading the administration of the dose. Conclusions: Below-target posaconazole concentrations were significantly more frequent in PPI users, graft-versus-host disease, and diarrhea. TDM seemed to be a helpful tool to identify low concentrations and to optimize posaconazole treatment.
Poxvirus ankyrin repeat proteins are a unique class of F-box proteins that associate with cellular SCF1 ubiquitin ligase complexes

F-box proteins direct the degradation of an extensive range of proteins via the ubiquitin-proteasome system. Members of this large family of proteins are typically bipartite. They recruit specific substrates through a substrate-binding domain and, via the F-box, link these to core components of a major class of ubiquitin ligases (SCF1). F-box proteins thus determine the specificity of SCF1-mediated ubiquitination. F-box-like motifs were recently detected in poxvirus ankyrin repeat (ANK) proteins but clear compositional differences to typical F-box proteins raise questions regarding the classification and function of the motif. Here we show that all five ANK proteins of a representative poxvirus, Orf virus, interact in vivo with core components of the SCF1 ubiquitin ligase complex. Interaction is dependent on the poxviral F-box-like motif and the adaptor subunit of the complex (SKP1). The viral protein does not block enzymatic activity of the complex. These observations identify the poxviral motif as a functional F-box. They also identify a new class of F-box that in contrast to cellular counterparts is truncated, has an extreme C-terminal location and is paired with an ANK protein-binding domain. ANK proteins constitute the largest family of poxviral proteins but their function and the significance of their abundance have remained an enigma. We propose that poxviruses use these unique ANK/F-box proteins to dictate target specificity to SCF1 ubiquitin ligases and thereby exploit the cell's ubiquitin-proteasome machinery.