Dynamics of a heparin-binding domain of VEGF(165) complexed with its inhibitor triamterene.

Vascular endothelial growth factor (VEGF), which has neurotrophic and neuroprotective effects in addition to its major role in angiogenesis, interacts with Aß and accumulates in the senile plaques of Alzheimer's disease (AD) patients' brains. It is known that Aß binds to the heparin-binding domain (HBD) of the 165-amino acid VEGF variant, VEGF(165). In this study, we showed that triamterene (Trm) inhibits VEGF--Aß interaction without affecting other biological activities of VEGF or Aß. We investigated the importance of structural and dynamic features of HBD for its molecular-recognition processes. The binding model of HBD and Trm was constructed based on measurements of chemical shift changes and docking study. The results showed that the loop region (S11-L17) and F18 at the beginning of the first ß-sheet in the HBD constitute the inhibitor binding site. The N1 atom of pteridine ring of Trm forms hydrogen bonding with backbone amide proton of R13, and the phenyl ring took part in a hydrophobic interaction with the aromatic ring of F18. To investigate the functional importance of the inherent structural flexibility of the HBD in VEGF, the dynamic properties of free HBD and HBD--Trm complex were assessed by measuring spin relaxation rates, and the backbone dynamics were investigated by model-free analysis. The residues in the disordered loop region of the N-terminus exhibited conformational exchanges in free HBD, and flexibility of this loop region decreased dramatically upon binding to Trm, suggesting that Aß as well as inhibitor may recognize these unique dynamic features of the HBD. Furthermore, C-terminal residues continued to exhibit slow conformational motions, even in the HBD--Trm complex, implying that these motions at the C-terminus of the HBD might be important for interactions with heparin molecules. The flexibility of HBD demonstrated here should be essential for VEGF function and interaction with other protein partners.

Structural flexibility and the positive charges are the key factors in bacterial cell selectivity and membrane penetration of peptoid-substituted analog of Piscidin 1.

Piscidin 1 (Pis-1) is a novel cytotoxic peptide with a cationic alpha-helical structure isolated from the mast cells of hybrid striped bass. In our previous study, we showed that Pis-1[PG] with a substitution of Pro(8) for Gly(8) in Pis-1 had higher bacterial cell selectivity than Pis-1. We designed peptoid residue-substituted peptide, Pis-1[NkG], in which Gly(8) of Pis-1 was replaced with Nlys (Lys peptoid residue). Pis-1[NkG] had higher antibacterial activity and lower cytotoxicity against mammalian cells than Pis-1 and Pis-1[PG]. We determined the tertiary structure of Pis-1[PG] and Pis-1[NkG] in the presence of DPC micelles by NMR spectroscopy. Both peptides had a three-turn helix in the C-terminal region and a bent structure in the center. Pis-1[PG] has a rigid bent structure at Pro(8) whereas Pis-1[NkG] existed as a dynamic equilibrium of two conformers with a flexible hinge structure at Nlys(8). Depolarization of the membrane potential of Staphylococcus aureus and confocal laser-scanning microscopy study revealed that Pis-1[NkG] effectively penetrated the bacterial cell membrane and accumulated in the cytoplasm, whereas Pis-1[PG] did not penetrate the membrane but remained outside or on the cell surface. Introduction of a lysine peptoid at position 8 of Pis-1 provided conformational flexibility and increased the positive charge at the hinge region; both factors facilitated penetration of the bacterial cell membrane and conferred bacterial cell selectivity on Pis-1[NkG].

Emphysema as a risk factor for the outcome of surgical resection of lung cancer.

It is unclear whether emphysema, regardless of airflow limitation, is a predictive factor associated with survival after lung cancer resection. Therefore, we investigated whether emphysema was a risk factor associated with the outcome after resection for lung cancer. This study enrolled 237 patients with non small cell lung cancer with stage I or II who had surgical removal. Patient outcome was analyzed based on emphysema. Emphysema was found in 43.4% of all patients. Patients with emphysema were predominantly men and smokers, and had a lower body mass index than the patients without emphysema. The patients without emphysema (n=133) survived longer (mean 51.2+/-3.0 vs. 40.6+/-3.1 months, P=0.042) than those with emphysema (n=104). The univariate analysis showed a younger age, higher FEV(1)/FVC, higher body mass index, cancer stage I, and a lower emphysema score were significant predictors of better survival. The multivariate analysis revealed a younger age, higher body mass index, and cancer stage I were independent parameters associated with better survival, however, emphysema was not. This study suggests that unfavorable outcomes after surgical resection of lung cancer should not be attributed to emphysema itself.

Overcoming artifacts from metallic orthopedic implants at high-field-strength MR imaging and multi-detector CT.

At magnetic resonance (MR) imaging and multidetector computed tomography (CT), artifacts arising from metallic orthopedic hardware are an obstacle to obtaining optimal images. Although various techniques for reducing such artifacts have been developed and corroborated by previous researchers, a new era of more powerful MR imaging and multidetector CT modalities has renewed the importance of a systematic consideration of methods for artifact reduction. Knowledge of the factors that contribute to artifacts, of related theories, and of artifact reduction techniques has become mandatory for radiologists. Factors that affect artifacts on MR images include the composition of the metallic hardware, the orientation of the hardware in relation to the direction of the main magnetic field, the strength of the magnetic field, the pulse sequence type, and other MR imaging parameters (mainly voxel size, which is determined by the field of view, image matrix, section thickness, and echo train length). At multidetector CT, the factors that affect artifacts include the composition of the hardware, orientation of the hardware, acquisition parameters (peak voltage, tube charge, collimation, and acquired section thickness), and reconstruction parameters (reconstructed section thickness, reconstruction algorithm used, and whether an extended CT scale was used). A comparison of images obtained with different hardware and different acquisition and reconstruction parameters facilitates an understanding of methods for reducing or overcoming artifacts related to metallic implants.

Morton neuroma: evaluated with ultrasonography and MR imaging.

OBJECTIVE: The purpose of this study was to compare the diagnostic accuracy of both ultrasonography (US) and magnetic resonance imaging (MRI) for the assessment of Morton neuroma. MATERIALS AND METHODS: Our study group was comprised of 20 neuromas from 17 patients, and the neuromas were confirmed on surgery following evaluation with US, MRI, or both US and MRI. The diagnostic criterion for Morton neuroma, as examined by US, was the presence of a round or ovoid, well-defined, hypoechoic mass. The diagnostic criterion, based on MR imaging, was a well defined mass with intermediate to low signal intensity on both the T1- and T2-weighted images. The retrospective comparison between the sonographic and MR images was done by two experienced radiologists working in consensus with the surgical and pathologic correlations. RESULTS: The detection rate of Morton neuroma was 79% for 14 neuromas from 11 patients who had undergone US followed by an operation. The detection rate was 76% for 17 neuromas from 15 patients who had undergone MRI and a subsequent operation. The mean size of the examined neuromas was 4.9 mm on the US images and it was 5.1 mm on the MRI studies. Ten neuromas (71%) were 5 mm or less as measured by US, and three neuromas were not detected, whereas on the MRI analysis, 10 neuromas (59%) were 5 mm or less and four neuromas were not visualized. Among the patients examined during postoperative follow-up, symptoms were completely relieved in 85% and the symptoms were partially relieved in 15%. CONCLUSION: US and MR imaging are comparable modalities with high detection rate for the evaluation of Morton neuroma.

Systematic review and meta-analysis of randomized controlled trials for the management of limited vertical height in the posterior region: short implants (5 to 8 mm) vs longer implants (> 8 mm) in vertically augmented sites.

PURPOSE: The aim of this study was to undertake a systematic review with meta-analysis on randomized controlled trials (RCTs) to compare the rates of survival, success, and complications of short implants to those of longer implants in the posterior regions. MATERIALS AND METHODS: Electronic literature searches were conducted through the MEDLINE (PubMed) and EMBASE databases to locate all relevant articles published between January 1, 1990, and April 30, 2013. Eligible studies were selected based on inclusion criteria, and quality assessments were conducted. After data extraction, meta-analyses were performed. RESULTS: In total, 539 dental implants (265 short implants [length 5 to 8 mm] and 274 control implants [length > 8 mm]) from four RCTs were included. The fixed prostheses of multiple short and control implants were all splinted. The mean follow-up period was 2.1 years. The 1-year and 5-year cumulative survival rates (CSR) were 98.7% (95% confidence interval [CI], 97.8% to 99.5%) and 93.6% (95% CI, 89.8% to 97.5%), respectively, for the short implant group and 98.0% (95% CI, 96.9% to 99.1%) and 90.3% (95% CI, 85.2% to 95.4%), respectively, for the control implant group. The CSRs of the two groups did not demonstrate a statistically significant difference. There were also no statistically significant differences in success rates, failure rates, or complications between the two groups. CONCLUSION: Placement of short dental implants could be a predictable alternative to longer implants to reduce surgical complications and patient morbidity in situations where vertical augmentation procedures are needed. However, only four studies with potential risk of bias were selected in this meta-analysis. Within the limitations of this meta-analysis, these results should be confirmed with robust methodology and RCTs with longer follow-up duration.

Backbone dynamics of an atypical orphan response regulator protein, Helicobacter pylori 1043.

An atypical orphan response regulator protein, HP1043 (HP-RR) in Helicobacter pylori, is proven to be essential for cell growth and does not require the well known phosphorelay scheme. HP-RR was identified as a symmetric dimer with two functional domains, an N-terminal regulatory domain (HP-RR(r)) and a C-terminal effector domain (HP-RR(e)). HP-RR is a new class of response regulator, as a phosphorylation-independent regulator. Previously, we have presented a detailed three-dimensional structure of HP-RR using NMR spectroscopy and X-ray crystallography. In this study, in order to understand the functional importance of flexibilities in HP-RR(r) and HP-RR(e), T1, T2, heteronuclear NOE experiments have been performed and backbone dynamics of HP-RR(r) and HP-RR(e) were investigated. HP-RR(r) is a symmetric dimer and the interface region, α4-ß5-α5 of dimer, showed high rigidity (high S (2) values). Site of rearrangements associated with phosphorylation of HP-RR(r) (Ser(75): R ex = 3.382, Ile(95): R ex = 5.228) showed slow chemical exchanges. HP-RR(e) is composed of three α-helices flanked on two sides by anti-parallel ß-sheets. Low order parameters as well as conformational exchanges in the centers of loop regions known as the DNA binding site and transcription site of HP-RR(e) suggested that flexibility of HP-RR(e) is essential for interaction with DNA. In conclusion, backbone dynamics information for HP-RR implies that structural flexibilities in HP-RR(r) are necessary for the phosphorylation site and the dynamic nature of HP-RR(e) is essential for the regulation of interaction between protein and DNA.

Interaction models of substrate peptides and beta-secretase studied by NMR spectroscopy and molecular dynamics simulation.

The formation of beta-amyloid peptide (Abeta) is initiated from cleavage of amyloid precursor protein (APP) by a family of protease, alpha-, beta-, and gamma-secretase. Sub W, a substrate peptide, consists of 10 amino acids, which are adjacent to the beta-cleavage site of wild-type APP, and Sub M is Swedish mutant with double mutations on the left side of the beta-cleavage site of APP. Sub W is a normal product of the metabolism of APP in the secretary pathway. Sub M is known to increase the efficiency of beta-secretase activity, resulting in a more specific binding model compared to Sub W. Three-dimensional structures of Sub W and Sub M were studied by CD and NMR spectroscopy in water solution. On the basis of these structures, interaction models of beta-secretase and substrate peptides were determined by molecular dynamics simulation. Four hydrogen bonds and one water-mediated interaction were formed in the docking models. In particular, the hydrogen bonding network of Sub M-BACE formed spread over the broad region of the active site of beta-secretase (P5-P3'), and the side chain of P2-Asn formed a hydrogen bond specifically with the side chain of Arg235. These are more favorable to the cleavage of Sub M by beta-secretase than Sub W. The two substrate peptides showed different tendency to bind to beta-secretase and this information may useful for drug development to treat and prevent Alzheimer's disease.

Role of magnetic resonance imaging in entrapment and compressive neuropathy--what, where, and how to see the peripheral nerves on the musculoskeletal magnetic resonance image: part 2. Upper extremity.

The diagnosis of nerve entrapment and compressive neuropathy has been traditionally based on the clinical and electrodiagnostic examinations. As a result of improvements in the magnetic resonance (MR) imaging modality, it plays not only a fundamental role in the detection of space-occupying lesions, but also a compensatory role in clinically and electrodiagnostically inconclusive cases. Although ultrasound has undergone further development in the past decades and shows high resolution capabilities, it has inherent limitations due to its operator dependency. We review the course of normal peripheral nerves, as well as various clinical demonstrations and pathological features of compressed and entrapped nerves in the upper extremities on MR imaging, according to the nerves involved. The common sites of nerve entrapment of the upper extremity are as follows: the brachial plexus of the thoracic outlet; axillary nerve of the quadrilateral space; radial nerve of the radial tunnel; ulnar nerve of the cubital tunnel and Guyon's canal; median nerve of the pronator syndrome, anterior interosseous nerve syndrome, and carpal tunnel syndrome. Although MR imaging can depict the peripheral nerves in the extremities effectively, radiologists should be familiar with nerve pathways, common sites of nerve compression, and common space-occupying lesions resulting in nerve compression in MR imaging.

Role of magnetic resonance imaging in entrapment and compressive neuropathy - what, where, and how to see the peripheral nerves on the musculoskeletal magnetic resonance image: part 1. Overview and lower extremity.

The diagnosis of nerve entrapment and compressive neuropathy has been traditionally based on the clinical and electrodiagnostic examinations. As a result of improvements in the magnetic resonance (MR) imaging modality, it plays not only a fundamental role in the detection of space-occupying lesions but also a compensatory role in clinically and electrodiagnostically inconclusive cases. Although ultrasound has undergone further development in the past decades and shows high resolution capabilities, it has inherent limitations due to its operator dependency. We review the general concepts that should be known to evaluate the entrapment and compressive neuropathy in MR imaging. We also review the course of normal peripheral nerves, as well as various clinical demonstrations and pathological features of compressed and entrapped nerves in the lower extremities on MR imaging, according to the nerves involved. The common sites of nerve entrapment of the lower extremity are as follows: sciatic nerve around the piriformis muscle; tibial nerve at the popliteal fossa and tarsal tunnel, common peroneal nerve around the fibular neck, and digital nerve near the metatarsal head. Although MR imaging can depict the peripheral nerves in the extremities effectively, radiologists should be familiar with nerve pathways, common sites of nerve compression, and common space-occupying lesions resulting in nerve compression in MR imaging.

Solution structure and cell selectivity of piscidin 1 and its analogues.

Piscidin 1 (Pis-1) is a novel cytotoxic peptide with a cationic alpha-helical structure that was isolated from the mast cells of hybrid striped bass [Silphaduang, U., and Noga, E. J. (2001) Nature 414, 268-269]. Pis-1 is not selective for bacterial versus mammalian cells. In the present study, to develop novel antibiotic peptides with selectivity for bacterial cells, we examined the effect of substituting two glycine residues, Gly8 and Gly13, with Ala or Pro on this peptide's structure and biological activities. The bacterial cell selectivity of the peptides decreased in the following order: Gly-->Pro analogues > Gly-->Pro/Ala analogues > Pis-1 > Gly-->Ala analogues. The antimicrobial and hemolytic activities and abilities to permeabilize the model phospholipid membranes were higher for Pis-1 with Gly or Pro at position 8 than for its counterparts with either Gly or Pro at position 13. We determined the tertiary structure of Pis-1 and its analogues in the presence of SDS micelles by NMR spectroscopy. We found that Pis-1 has an alpha-helical structure from Phe2 to Thr21. Also, Pis-1 AA (Gly8, Gly13-->Ala8, Ala13) with higher antibacterial and hemolytic activity than Pis-1 has a stable alpha-helical structure from Phe2 to Thr21. Pis-1 PG (Gly-->Pro8) with bacterial cell selectivity has a hinge structure at Pro8, which provides flexibility in piscidin, followed by a three-turn helix from Val10 to Gly22 in the C-terminal region. Taken together, our results demonstrate that the conformational flexibility provided by introduction of a Pro at position 8, coupled with the primary anchoring of phenylalanines and histidines in the N-terminus to the cell membrane and the optimal length of the C-terminal amphipathic alpha-helix, are the critical factors that confer antibacterial activity and bacterial cell selectivity to Pis-1 PG. Pis-1 PG may be a good candidate for the development of a new drug with potent antibacterial activity but without cytotoxicity.

Chronic tibiofibular syndesmosis injury of ankle: evaluation with contrast-enhanced fat-suppressed 3D fast spoiled gradient-recalled acquisition in the steady state MR imaging.

PURPOSE: To retrospectively determine the accuracy of coronal contrast material-enhanced fat-suppressed three-dimensional (3D) fast spoiled gradient-recalled acquisition in the steady state (SPGR) magnetic resonance (MR) imaging, as compared with that of routine transverse MR imaging, in the assessment of distal tibiofibular syndesmosis injury, with arthroscopy as the reference standard. MATERIALS AND METHODS: The review board of the College of Medicine in Yonsei University approved this study; informed consent was waived. The study group comprised 45 patients (26 men, 19 women; mean age, 32.1 years; range, 18-58 years) with a chronic ankle injury who had undergone MR imaging and arthroscopic surgery. Three independent readers retrospectively reviewed the two sets of MR images (one set of gadolinium-enhanced 3D fast SPGR images and one set of routine T1-, T2-, and intermediate-weighted images). Scores from 1 to 5 in increasing order of the probability of injury were assigned to both sets. Arthroscopy was the reference standard. Syndesmotic recess height was measured on contrast-enhanced images. The two sets of images were compared for diagnostic performance with receiver operating characteristic (ROC) analysis. Dissection and histologic examination of six cadaveric ankles was performed to assess the syndesmotic area and ascertain the enhancing structure at MR imaging. RESULTS: At arthroscopy, syndesmotic injury was found in 24 ankles but not in 21 ankles. Areas under the ROC curve were significantly higher for the contrast-enhanced images (P<.05). The contrast-enhanced set showed higher accuracy, sensitivity, and specificity compared with the routine set for the assessment of syndesmosis injury. Mean syndesmotic recess height was significantly greater (P<.05) in patients with syndesmotic injury. Dissection and histologic examination revealed a highly vascular synovial fold in the syndesmotic area that is expected to enhance at MR imaging. CONCLUSION: In the assessment of chronic syndesmosis injury, coronal gadolinium-enhanced fat-suppressed 3D fast SPGR MR images were more sensitive, specific, and accurate than routine MR images.

Structure of an atypical orphan response regulator protein supports a new phosphorylation-independent regulatory mechanism.

Two-component signal transduction systems, commonly found in prokaryotes, typically regulate cellular functions in response to environmental conditions through a phosphorylation-dependent process. A new type of response regulator, hp1043 (HP-RR) from Helicobacter pylori, has been recently identified. HP-RR is essential for cell growth and does not require the well known phosphorelay scheme. Unphosphorylated HP-RR binds specifically to its own promoter (P(1043)) and autoregulates the promoter of the tlpB gene (P(tlpB)). We have determined the structure of HP-RR by NMR and x-ray crystallography, revealing a symmetrical dimer with two functional domains. The molecular topology resembles that of the OmpR/PhoB subfamily, however, the symmetrical dimer is stable even in the unphosphorylated state. The dimer interface, formed by three secondary structure elements (alpha4-beta5-alpha5), resembles that of the active, phosphorylated forms of ArcA and PhoB. Several conserved residues of the HP-RR dimeric interface deviate from the OmpR/PhoB subfamily, although there are similar salt bridges and hydrophobic patches within the interface. Our findings reveal how a new type of response regulator protein could function as a cell growth-associated regulator in the absence of post-translational modification.

Anterior-inferior labral lesions of recurrent shoulder dislocation evaluated by MR arthrography in an adduction internal rotation (ADIR) position.

PURPOSE: To introduce and evaluate the usefulness of the adduction internal rotation (ADIR) position in MR arthrography for discriminating the following subtypes of Bankart lesions: classic Bankart lesions, anterior labroligamentous periosteal sleeve avulsion (ALPSA) lesions, and Perthes lesions. MATERIALS AND METHODS: The study group consisted of 25 patients who had been referred for MR arthrography of the shoulder and then underwent arthroscopy due to recurrent episodes of shoulder dislocation. MR arthrography was performed in three different positions: neutral, abduction external rotation (ABER), and ADIR. The authors evaluated the statistical significance of the method's discriminative diagnostic ability for the subtypes of Bankart lesions according to the position changes. RESULTS: Twenty-three patients had the following single or complex labral lesions: classic Bankart (N = 8), ALPSA (N = 3), complex classic Bankart/ALPSA (N = 8), and complex ALPSA/Perthes (N = 4). Two patients had no visible anteroinferior labral lesions. The performance of ADIR positioning for differentiating ALPSA lesions was superior to the neutral or ABER positioning. The difference of the discriminative lesion detection ability was statistically significant (P <.01) only for detecting ALPSA lesions in the ADIR position. CONCLUSION: MR arthrography in the ADIR position provides high accuracy for the diagnosis of ALPSA lesions, and complements routine MR arthrography when used to diagnose labroligamentous lesions in patients with recurrent shoulder dislocations.