Agenesis of the hemidiaphragm: a rare presentation in an adult.

Unilateral diaphragmatic agenesis is a rare finding in adult patients. We report a case of unilateral agenesis of diaphragm in a 22-year-old male patient to highlight the fact that a rare entity of agenesis of diaphragm can have a misleading presentation in adulthood due to both pulmonary and abdominal symptoms.

The use of a powdered polysaccharide hemostat (HemoStase) in live donor nephrectomies controls bleeding and reduces postoperative complications.

OBJECTIVE: This study assesses the use of microporous hemospheres (hemoStase) procedures to control bleeding and reduce associated unfavorable sequelae in live donor nephrectomy (LDN). METHODS: Forty-four consecutive patients who underwent LDN between January 2009 and August 2009 were included in this prospective study. HemoStase (CryoLife, Inc), Kennesaw, Georgia) was used topically in the kidney bed to avoid bleeding. We recorded intraoperative and postoperative bleeding control, re-exploration to control bleeding, surgical site fluid collection, infection, and postoperative wound complications. RESULTS: Hemostasis was achieved in all 44 (100%) patients, none of whom experienced postoperative bleeding, fluid collection, infection or required re-exploration of the surgical site. CONCLUSION: HemoStase is a safe, facile hemostatic agent that effectively controls bleeding and reduces associated postoperative complications in LDN cases.

A fast and safe living donor "finger-assisted" nephrectomy technique: results of 359 cases.

OBJECTIVE: To determine operative parameters and complications, using a modified approach to mini-incision living donor nephrectomy. METHODS: Three hundred fifty-nine consecutive living donor procedures were performed between October 2000 and November 2008 using the finger-assisted, mini-incision living donor nephrectomy. Patient demographics, intraoperative parameters, and postoperative complications were prospectively recorded, including operative time, blood loss, incision length, warm ischemia time, and intraoperative adverse events. RESULTS: Mean donor age was 44.2 +/- 12.3 years (range, 21-75 years), with an average body mass index of 28.2 +/- 5.3 kg/m(2) (range, 17.1-44.9 kg/m(2)). Right-sided donor nephrectomies were performed on 23 patients (6%), and 41 donors (11%) were found to have multiple renal arteries. Median incision length was 6.8 cm (range, 3.5-15 cm). Average operative time was 117 minutes (range, 50-265 minutes), with a median blood loss of 109 mL (range, 20-500 mL) and an average warm ischemia time of 4.5 minutes (range, 1.5-10 minutes). Four patients (1%) required perioperative blood transfusions. There were no other intraoperative complications, no patients required reexploration, and there were no donor deaths. Thirteen patients (4%) developed minor postoperative complications, including two incisional herniae, but no patients developed chronic wound pain, over a median follow-up period of 19 months (range, 2-97 months). CONCLUSION: This prospective series demonstrated that a modified approach to open mini-incision nephrectomy can result in a smaller incision length while maintaining patient safety, with few postoperative complications.

Scattering of the fundamental shear horizontal guided wave by a part-thickness crack in an isotropic plate.

The interaction of the fundamental shear horizontal (SH0) guided mode with part-thickness cracks in an isotropic plate is studied as an extension within the context and general framework of previous work ["Short range scattering of the fundamental shear horizontal guided wave mode normally incident at a through thickness crack in an isotropic plate," J. Acoust Soc. Am. 122, 1527-1538 (2007); "Angular influence on scattering when the fundamental shear horizontal guided wave mode is incident at a through-thickness crack in an isotropic plate," J. Acoust. Soc. Am. 124, 2021-2030 (2008)] by the authors with through-cracks. The symmetric incidence case where the principal direction of the incident beam bisects the crack face at 90 degrees is studied using finite element simulations validated by experiments and analysis, and conclusions are inferred for general incidence angles using insights obtained with the through-thickness studies. The influence of the crack length and the monitoring distance on the specular reflection is first examined, followed by a study of the angular profile of the reflected field. With each crack length considered, the crack depth and operating frequencies are varied. For all crack depths studied, the trend of the results is identical to that for the corresponding through-thickness case and the values differ only by a frequency dependent scale factor. Theoretical analysis is used to understand the physical basis for such behavior and estimates are suggested for the scale factor--exact for the high-frequency scattering regime and empirical for the medium- and low-frequency regimes.

Angular influence on the scattering of fundamental shear horizontal guided waves by a through-thickness crack in an isotropic plate.

The angular influence on the scattering of cylindrical-crested waves of the fundamental shear horizontal (SH0) guided mode by through-thickness cracks in an isotropic plate is studied in the context of array imaging using ultrasonic guided waves. Finite element simulations are used to obtain trends which are subject to analytical study and experimental confirmation. The influence of the incidence angle on reflection behavior is first studied in terms of two complementary cases, that of normal incidence and that of specular reflection at various oblique incidence angles. The normal incidence study suggests that for a given incidence angle, the peak reflection is concentrated around the specular direction, while the oblique incidence studies show that maximum specular reflection occurs in the case of normal incidence. The variation of diffraction with both the angle of incidence and that of monitoring is then taken up and this shows that when the first diffraction from the crack edges can be separated, its angular dependence can be obtained from literature on similar bulk elastic wave scattering problems.

Short range scattering of the fundamental shear horizontal guided wave mode normally incident at a through-thickness crack in an isotropic plate.

Interaction of the fundamental shear horizontal mode with through-thickness cracks in an isotropic plate is studied in the context of low frequency array imaging for ultrasonic guided wave nondestructive evaluation with improved resolution. Circular wave fronts are used and the symmetric case where a line from the wave source bisects the crack face normally is considered. Finite element simulations are employed to obtain trends subject to analytical and experimental validation. The influence of the crack length and of the location of source and measurement positions on the specular reflection from the crack face is first examined. These studies show that low frequency short range scattering is strongly affected by diffraction phenomena, leading to focusing of energy by the crack in the backscatter direction. Study of the diffraction from the crack edges reveals contributions due to a direct diffraction at the edges and multiple reverberations across the crack length. A simple diffraction model is shown to adequately represent cracks up to moderate lengths, providing an easy means of estimating the far field of the waves. The presence of multiple diffraction components is quantitatively established and surface waves on the crack face are identified as equivalent to low frequency symmetric modes of rectangular ridge waveguides.

Growth temperature dependence of optical modal gain and loss in GaN:Eu active medium on Si.

The dependence of optical modal gain and loss on GaN:Eu growth temperature is reported. GaN:Eu thin films were grown on Si substrates with AlGaN transition and cladding layers at temperatures ranging from 600 degrees C to 850 degrees C. The modal gain and loss in the GaN:Eu layer were a strong function of the optically active Eu atomic concentration and of the interface quality between the active layer and the top cladding layer, which in turn depended on the growth temperature. Optimum optical properties of maximum modal gain of ~ 100 cm(-1) and minimum loss of ~ 46 cm(-1) were obtained for growth at 800 degrees C.

Effect of Cassia auriculata Linn. root extract on cisplatin and gentamicin-induced renal injury.

The ethanol extract of the roots of Cassia auriculata was studied for its nephroprotective activity in cisplatin- and gentamicin-induced renal injury in male albino rats. In the cisplatin model, the extract at doses of 300 and 600 mg/kg body wt. reduced elevated blood urea and serum creatinine and normalized the histopathological changes in the curative regimen. In the gentamicin model, the ethanol extract at a dose of 600 mg/kg body wt. reduced blood urea and serum creatinine effectively in both the curative and the preventive regimen. The extract had a marked nitric oxide free-radical-scavenging effect. The findings suggest that the probable mechanism of nephroprotection by C. auriculata against cisplatin- and gentamicin-induced renal injury could be due to its antioxidant and free-radical-scavenging property.

Clinicopathological study of thymomas--correlation of histologic subtype to myasthenia gravis and prognosis.

Thymoma is the most common primary tumor of anterior superior mediastinum. Sixty cases of thymomas over a 12 year period were analysed and the histologic subtype, according to Marino and Muller-Hermilink, classification was correlated with presence or absence of myasthenia gravis (MG) and capsular invasion. Thirty four patients had myasthenia gravis associated with thymoma and there was one case of pure red cell aplasia. There were 3 (1) predominantly cortical, 28 (20) cortical, 12 (9) mixed, 16 (4) medullary thymomas and 1 (0) thymic carcinoma (Figures in parenthesis indicate number of cases associated with MG). Capsular invasion was seen in 25 cases. Association with myasthenia gravis and capsular invasion were seen predominantly in cortical and mixed thymomas which were also associated with aggressive behaviour.

Structure of a BRCA1-BARD1 heterodimeric RING-RING complex.

The RING domain of the breast and ovarian cancer tumor suppressor BRCA1 interacts with multiple cognate proteins, including the RING protein BARD1. Proper function of the BRCA1 RING domain is critical, as evidenced by the many cancer-predisposing mutations found within this domain. We present the solution structure of the heterodimer formed between the RING domains of BRCA1 and BARD1. Comparison with the RING homodimer of the V(D)J recombination-activating protein RAG1 reveals the structural diversity of complexes formed by interactions between different RING domains. The BRCA1-BARD1 structure provides a model for its ubiquitin ligase activity, illustrates how the BRCA1 RING domain can be involved in associations with multiple protein partners and provides a framework for understanding cancer-causing mutations at the molecular level.

Tetralogy of Fallot with flap valve ventricular septal defect producing suprasystemic right ventricular pressure: echocardiographic observations.

Tetralogy of Fallot with restrictive ventricular septal defect and suprasystemic right ventricular pressure is an uncommon anomaly with high mortality rate. Very few such cases have been reported pre-operatively. This report describes echocardiographic features of five patients with restrictive ventricular septal defect due to flap valve in tetralogy of Fallot. In all the five cases, accessory or excessive tricuspid valve tissue obstructed the defect. Recognition of restrictive ventricular septal defect in tetralogy of Fallot is important because it appears to have poor prognosis.

Prediction and structural characterization of an independently folding substructure in the src SH3 domain.

Previous studies of the conformations of peptides spanning the length of the alpha-spectrin SH3 domain suggested that SH3 domains lack independently folding substructures. Using a local structure prediction method based on the I-sites library of sequence-structure motifs, we identified a seven residue peptide in the src SH3 domain predicted to adopt a native-like structure, a type II beta-turn bridging unpaired beta-strands, that was not contained intact in any of the SH3 domain peptides studied earlier. NMR characterization confirmed that the isolated peptide, FKKGERL, adopts a structure similar to that adopted in the native protein: the NOE and 3JNHalpha coupling constant patterns were indicative of a type II beta-turn, and NOEs between the Phe and the Leu side-chains suggest that they are juxtaposed as in the prediction and the native structure. These results support the idea that high-confidence I-sites predictions identify protein segments that are likely to form native-like structures early in folding.

Solvent exchange rates of side-chain amide protons in proteins.

Solvent exchange rates and temperature coefficients for Asn/Gln side-chain amide protons have been measured in Escherichia coli HPr. The protons of the eight side-chain amide groups (two Asn and six Gln) exhibit varying exchange rates which are slower than some of the fast exchanging backbone amide protons. Differences in exchange rates of the E and Z protons of the same side-chain amide group are obtained by measuring exchange rates at pH values > 8. An NOE between a side-chain amide proton and a bound water molecule was also observed.

Phosphorylation on histidine is accompanied by localized structural changes in the phosphocarrier protein, HPr from Bacillus subtilis.

The histidine-containing protein (HPr) of bacterial phosphoenolpyruvate:sugar phosphotransferase system (PTS) serves a central role in a series of phosphotransfer reactions used for the translocation of sugars across cell membranes. These studies report the high-definition solution structures of both the unphosphorylated and histidine phosphorylated (P-His) forms of HPr from Bacillus subtilis. Consistent with previous NMR studies, local conformational adjustments occur upon phosphorylation of His 15, which positions the phosphate group to serve as a hydrogen bond acceptor for the amide protons of Ala 16 and Arg 17 and to interact favorably with the alpha-helix macrodipole. However, the positively charged side chain of the highly conserved Arg 17 does not appear to interact directly with phospho-His 15, suggesting that Arg 17 plays a role in the recognition of other PTS enzymes or in phosphotransfer reactions directly. Unlike the results reported for Escherichia coli P-His HPr (Van Nuland NA, Boelens R, Scheek RM, Robillard GT, 1995, J Mol Biol 246:180-193), our data indicate that phosphorylation of His 15 is not accompanied by adoption of unfavorable backbone conformations for active site residues in B. subtilis P-Ser HPr.

NMR chemical shift perturbation mapping of DNA binding by a zinc-finger domain from the yeast transcription factor ADR1.

Mutagenesis studies have revealed that the minimal DNA-binding domain of the yeast transcription factor ADR1 consists of two Cys2-His2 zinc fingers plus an additional 20 residues proximal and N-terminal to the fingers. We have assigned NMR 1H, 15N, and 13C chemical shifts for the entire minimal DNA-binding domain of ADR1 both free and bound to specific DNA. 1H chemical shift values suggest little structural difference between the zinc fingers in this construct and in single-finger constructs, and 13C alpha chemical shift index analysis indicates little change in finger structure upon DNA binding. 1H chemical shift perturbations upon DNA binding are observed, however, and these are mapped to define the protein-DNA interface. The two zinc fingers appear to bind DNA with different orientations, as the entire helix of finger 1 is perturbed, while only the extreme N-terminus of the finger 2 helix is affected. Furthermore, residues N-terminal to the first finger undergo large chemical shift changes upon DNA binding suggesting a role at the protein-DNA interface. A striking correspondence is observed between the protein-DNA interface mapped by chemical shift changes and that previously mapped by mutagenesis.

Demonstration of protein-protein interaction specificity by NMR chemical shift mapping.

Chemical shift mapping is becoming a popular method for studying protein-protein interactions in solution. The technique is used to identify putative sites of interaction on a protein surface by detecting chemical shift perturbations in simple (1H, 15N)-HSQC NMR spectra of a uniformly labeled protein as a function of added (unlabeled) target protein. The high concentrations required for these experiments raise questions concerning the possibility for non-specific interactions being detected, thereby compromising the information obtained. We demonstrate here that the simple chemical shift mapping approach faithfully reproduces the known functional specificities among pairs of closely related proteins from the phosphoenolpyruvate:sugar phosphotransferase systems of Escherichia coli and Bacillus subtilis.

Influence of N-cap mutations on the structure and stability of Escherichia coli HPr.

This paper describes the effect of N-capping substitutions on the structure and stability of histidine-containing protein (HPr). We have used NMR spectroscopy and conformational stability studies to quantify changes in local and global free energy due to mutagenesis at Ser46, the N-cap for helix B in HPr. Previous NMR studies suggested that helix B of Escherichia coli HPr is dynamic as judged by the rate of exchange of amide protons with solvent. Ser46 was chosen because it is the site of regulatory phosphorylation in HPrs from Gram-positive bacteria, and mutation of this residue to an aspartic acid (S46D) in E. coli HPr (Gram-negative) also makes it a poor substrate in the bacterial phosphoenolpyruvate: sugar phosphotransferase system. Therefore, to understand the mechanism of inactivation of E. coli S46D HPr, as well as the effect of mutagenesis on protein stability, we have characterized three mutants of E. coli HPr: Ser46 has been mutated to an Asp, Asn, and Ala in S46D, S46N, and S46A HPrs, respectively. The results indicate that these N-cap replacements have a marked influence on helix B stability. The effect of mutagenesis on local stability is correlated to global unfolding of HPr. The ability of amino acids to stabilize helix B is Asp > Asn > Ser > Ala. In addition, since there are neither large-scale conformational changes nor detectable changes in the active site of S46D HPr, it is proposed that the loss of phosphotransfer activity of S46D HPr is due to unfavorable steric and/or electrostatic interactions of the Asp with enzyme I of the PTS.

Phosphorylation of serine-46 in HPr, a key regulatory protein in bacteria, results in stabilization of its solution structure.

The serine-phosphorylated form of histidine-containing protein (HPr), a component of the phosphoenolpyruvate:sugar phosphotransferase system from Bacillus subtilis, has been characterized by NMR spectroscopy and solvent denaturation studies. The results indicate that phosphorylation of Ser 46, the N-cap of alpha-helix-B, does not cause a conformational change but rather stabilizes the helix. Amide proton exchange rates in helix-B are decreased and phosphorylation stabilizes the protein to solvent and thermal denaturation, with a delta delta G of 0.7-0.8 kcal mol-1. A mutant in which Ser 46 is replaced by aspartic acid shows a similar stabilization, indicating that an electrostatic interaction between the negatively charged groups and the helix macrodipole contributes significantly to the stabilization.

Structural consequences of histidine phosphorylation: NMR characterization of the phosphohistidine form of histidine-containing protein from Bacillus subtilis and Escherichia coli.

The bacterial phosphoenolpyruvate:sugar phosphotransferase system involves a series of reactions in which phosphoprotein intermediates are formed. Histidine-containing protein (HPr) is phosphorylated on the N delta 1 position of the imidazole ring of His15 by enzyme I and acts as a phosphoryl donor to the sugar-specific enzymes IIA. The structure of phosphorylated HPr from Bacillus subtilis, primarily, and from Escherichia coli has been studied by nuclear magnetic resonance (NMR) spectroscopy. Phosphorylation of His15 results in large downfield shifts in amide proton and nitrogen resonances for residues 16 and 17 but results in only modest or no shifts in other backbone resonances. The exchange rates of these two amide groups are decreased more than 10-fold upon phosphorylation. Analysis of the coupling constants 3JNH alpha revealed no significant changes throughout the protein, indicating that backbone phi dihedral angles do not change detectably. 3J alpha beta and 3JN beta patterns determined from P.E.COSY and HNHB spectra, respectively, revealed a change in one side chain, that of conserved Arg17. Analysis of NOESY spectra revealed a limited number of changes in NOEs involving protons in Ser12, His15, Arg71, and Pro18 in B. subtilis HPr. The NMR results indicate that the Arg17 side chain becomes limited in its conformational range in the phosphorylated protein, taking on a conformation that points its guanidinium group toward the phosphoryl group on His15. In addition, the tautomeric and ionization states of His15 have been determined using 15N and 31P NMR. At neutral pH, the imidazole is predominantly in the protonated form and the phosphoryl group is in the dianionic form in P-His15. Altogether, the results indicate that phosphorylation of His15 yields only a local effect on the protein's structure. The data are consistent with a small change in the disposition of the histidine side chain, allowing phosphoryl group oxygens to serve as hydrogen bond acceptors for the amide protons of residues Ala16 and Arg17, which constitute the first two residues of an alpha-helix. Thus, similar to many proteins that bind phosphoryl moieties noncovalently, the phosphoryl group in P-His15-HPr is situated to allow for a favorable electrostatic interaction at the N-terminal end of an alpha-helix.