Optical nonlocalities and additional waves in epsilon-near-zero metamaterials.

We analyze the optical properties of plasmonic nanorod metamaterials in the epsilon-near-zero regime and show, both theoretically and experimentally, that the performance of these composites is strongly affected by nonlocal response of the effective permittivity tensor. We provide the evidence of interference between main and additional waves propagating in the room-temperature nanorod metamaterials and develop an analytical description of this phenomenon. Additional waves are present in the majority of low-loss epsilon-near-zero structures and should be explicitly considered when designing applications of epsilon-near-zero composites, as they represent a separate communication channel.

A comparison between red clover silage and grass silage feeding on fatty acid composition, meat stability and sensory quality of the M. Longissimus muscle of dairy cull cows.

Sixteen Holstein-Friesian dairy cull cows were offered either ad libitum grass (G) or red clover (RC) silage for 12 weeks. Dry matter (DM), total nitrogen, and pH was higher for the RC than the G silage and organic matter, water-soluble carbohydrate, fibre, DM digestibility, ammonia-N, vitamin E and acetic acid higher for the G silage (P<0.05). Fatty acid compositions were different (P<0.05) with G silage having higher levels of C12:0, C14:0, C16:1cis-9, C18:3n-3 and total fatty acids whereas RC had higher levels of C18:0, C18:2n-6 and C20:0. Daily liveweight gain was high and not different between groups (average 1.22 kg/d). Body condition score and back fat thickness at slaughter along with conformation, fat grade and slaughter weight were not different between groups. Animals offered the G silage produced larger M. longissimus length (P<0.01) and a trend (P<0.1) for width. RC fed animals had higher proportions of C18:3n-3 (P<0.001), total n-3 fatty acids (P<0.01) and total PUFA compared to animals offered the G silage despite greater intakes of these fatty acids on G (P<0.001). Vitamin E concentration and stability of aged meat during simulated retail display, were lower (P<0.05) from animals offered RC than animals offered G. Shear force and ultimate pH, however were not different in steaks and sensory attributes were similar; the only difference being a higher score (P<0.01) for fishy in the RC steaks. The results suggest that feeding high DMD silage to dairy cull cows can result in a high standard of finish and liveweight gain. Furthermore RC silage as opposed to G silage can increase the beneficial fatty acid profile of the resultant meat.

Towards nonlinear plasmonic devices based on metallic nanorods.

The nonlinear optical properties of gold nanorod arrays have been studied with a two-colour continuously working (CW) pump-probe spectroscopy. The hybridization of the nanorod arrays with a nonlinear polymer (poly-3BCMU) has been investigated for the purposes of active photonic component design. The sensitivity of the plasmonic resonances of the hybrid nanostructured system to pump-induced changes in the refractive index of the polymer surroundings has been used to control the optical extinction of the array. Both reversible and non-reversible behaviour has been observed reflecting the combined effects from the third-order nonlinear response of the hybrid metallo-dielectric system and post-photopolymerization triggered reactions in the polymer.

Optical transmission measurements of silver, silver-gold alloy and silver-gold segmented nanorods in thin film alumina.

Silver nanorods have been grown by electrodeposition into thin film porous alumina. Transmission measurements show two peaks related to the transverse and longitudinal resonance of the nanorods. The behaviour of the longitudinal resonance peak is found to vary with nanorod length and the spectral position to depend on nanorod diameter. As the distance between the nanorods is decreased a small blue-shift of the longitudinal peak is observed. Depositing a small gold cap on top of the silver nanorods causes a red-shift of the longitudinal peak whilst, conversely, the longitudinal peak of gold nanorod arrays is comparatively insensitive to the deposition of a silver cap. Gold-silver alloy nanorods were also deposited from a mixed salt bath and a linear dependence of the transverse peak position on alloy composition was observed.

Poor outcome following epidural abscess complicating epidural analgesia for labour.

Abscess is a rare complication of epidural analgesia that may occur post-partum. A case is described where a lumbar epidural abscess initially presented with back and leg pain 7 days after seemingly uneventful block for labour and forceps delivery of twins. Definitive diagnosis of abscess was delayed until 5 days later when neurological sequelae became apparent. Despite surgical evacuation, recovery was delayed and incomplete. The importance of prompt diagnosis and the involvement of anaesthetic personnel are emphasised.

Rotations and rotation matrices.

In molecular replacement, a model, described as a list of orthogonal coordinates, is to be moved into a new position and orientation. The orthogonal coordinate system also has to be related to the crystallographic system, which may not be orthogonal, and crystallographic symmetry must be considered, which applies to coordinates expressed as fractions of the unit cell. Elementary properties of rotation matrices and their representation as polar or Eulerian angles are discussed.

Alterations in cerebral potentials evoked by rectal distension in irritable bowel syndrome.

OBJECTIVE: Central nervous system correlates of the visceral hyperalgesia documented in patients with irritable bowel syndrome are limited. Reproducible cerebral evoked potentials can be recorded in response to rhythmic balloon distension of the rectum in healthy adults. Irritable bowel syndrome patients and healthy subjects were studied to compare the characteristics of mechanically-evoked rectal cerebral potentials obtained during fasting and after the ingestion of a standard meal. METHODS: Twenty-two pairs of age-matched healthy female subjects and female irritable bowel syndrome patients were studied. Cerebral evoked potentials were recorded in response to rhythmic rectal distension (two distension series each of 100 repetitions at 0.8 hertz); cerebral evoked potential recordings were repeated after a 1000 kcal (46% fat) liquid meal. Trait and state anxiety questionnaires were also completed. RESULTS: Compared to healthy subjects, irritable bowel syndrome patients demonstrated higher prevalence of cerebral evoked potential early peaks (latency < 100 ms) postprandially, and uniformly shorter cerebral evoked potential latencies both before and after feeding. CONCLUSION: These findings provide further objective evidence for defective visceral afferent transmission in irritable bowel syndrome patients.

Influence of patient and donor cytokine genotypes on renal allograft rejection: evidence from a single centre study.

Cytokines are key immune mediators and it has been suggested that cytokine gene polymorphisms affecting expression influence rejection or tolerance. This study sought to examine this hypothesis with the aim of identifying predictive genotype markers for rejection. The study group consisted of 120 consecutive first cadaveric recipient-donor pairs transplanted at a single centre, between 1994 and 1997. PCR utilising sequence-specific primers (SSP) methodology was optimised for genotyping recipient and donor DNA for the following polymorphisms: tumour necrosis factor (TNF) -alpha (-308, G/A), interleukin (IL)-10 (-1082, G/A), IL-4 (-590, C/T), transforming growth factor (TGF) -beta1 (+915, G/C). Recipient-donor pairs were divided into rejectors (n = 28) and non-rejectors (n = 92). Each group was further stratified according to number of rejection episodes and HLA-DR mismatching. Recipient-donor pairs both lacking the IL-4*T allele (recipient low producer/donor low producer) were significantly increased in the rejector group (P = 0.02). Also, the combination of recipient IL-10*A negative/donor IL-10*A positive (recipient high producer/donor low producer), was significantly decreased in multiple rejectors (P = 0.04). No significant associations were detected between TNF-alpha and TGF-beta1, and rejection. This study suggests that the combination of recipient-donor IL-4 and IL-10 genotypes may be important in renal transplantation outcome. The results appear to corroborate the protective role of both of these cytokines, possibly due to their ability to suppress inflammation. However, due to conflicting results from this and other studies, a multi-centre collaborative study may be required to determine whether cytokine genotypes are significant, independent predictors of renal allograft rejection.

A third specificity-determining site in mu 2 adaptin for sequences upstream of Yxx phi sorting motifs.

Internalization signals of the Yxx phi type (phi = bulky hydrophobic side chain) interact with the mu 2 chain of AP-2 adaptors. Internalization activity is intolerant of non-conservative substitution of either the tyrosine or the phi side chains, which bind to hydrophobic pockets in mu 2 adaptin in a conformation described as 'a two pinned plug into a socket'. P-selectin, a type I transmembrane protein, contains the Yxx phi-like sequence YGVF in its cytoplasmic domain, but substitution of either the tyrosine or phenylalanine with alanine in the full-length protein causes only small changes in the rate of endocytosis. It is shown here that the sequence YGVF contained within a peptide corresponding to the 17 COOH-terminal amino acids of P-selectin binds to mu 2 adaptin in the same fashion previously seen for other Yxx phi motifs. In addition, the P-selectin peptide binds to a third hydrophobic pocket in mu 2 adaptin through a leucine at position Y-3 in the peptide. This structure suggests that some sequences can function as a 'three pinned plug', in which internalization activity is not critically dependent on any one of the three interacting side chains.

Simultaneous binding of PtdIns(4,5)P2 and clathrin by AP180 in the nucleation of clathrin lattices on membranes.

Adaptor protein 180 (AP180) and its homolog, clathrin assembly lymphoid myeloid leukemia protein (CALM), are closely related proteins that play important roles in clathrin-mediated endocytosis. Here, we present the structure of the NH2-terminal domain of CALM bound to phosphatidylinositol-4,5- bisphosphate [PtdIns(4,5)P2] via a lysine-rich motif. This motif is found in other proteins predicted to have domains of similar structure (for example, Huntingtin interacting protein 1). The structure is in part similar to the epsin NH2-terminal (ENTH) domain, but epsin lacks the PtdIns(4,5)P2-binding site. Because AP180 could bind to PtdIns(4,5)P2 and clathrin simultaneously, it may serve to tether clathrin to the membrane. This was shown by using purified components and a budding assay on preformed lipid monolayers. In the presence of AP180, clathrin lattices formed on the monolayer. When AP2 was also present, coated pits were formed.

The structural basis for the recognition of acetylated histone H4 by the bromodomain of histone acetyltransferase gcn5p.

The bromodomain is an approximately 110 amino acid module found in histone acetyltransferases and the ATPase component of certain nucleosome remodelling complexes. We report the crystal structure at 1.9 A resolution of the Saccharomyces cerevisiae Gcn5p bromodomain complexed with a peptide corresponding to residues 15-29 of histone H4 acetylated at the zeta-N of lysine 16. We show that this bromodomain preferentially binds to peptides containing an N:-acetyl lysine residue. Only residues 16-19 of the acetylated peptide interact with the bromodomain. The primary interaction is the N:-acetyl lysine binding in a cleft with the specificity provided by the interaction of the amide nitrogen of a conserved asparagine with the oxygen of the acetyl carbonyl group. A network of water-mediated H-bonds with protein main chain carbonyl groups at the base of the cleft contributes to the binding. Additional side chain binding occurs on a shallow depression that is hydrophobic at one end and can accommodate charge interactions at the other. These findings suggest that the Gcn5p bromodomain may discriminate between different acetylated lysine residues depending on the context in which they are displayed.

The structure and function of the beta 2-adaptin appendage domain.

The heterotetrameric AP2 adaptor (alpha, beta 2, mu 2 and sigma 2 subunits) plays a central role in clathrin-mediated endocytosis. We present the protein recruitment function and 1.7 A resolution structure of its beta 2-appendage domain to complement those previously determined for the mu 2 subunit and alpha appendage. Using structure-directed mutagenesis, we demonstrate the ability of the beta 2 appendage alone to bind directly to clathrin and the accessory proteins AP180, epsin and eps15 at the same site. Clathrin polymerization is promoted by binding of clathrin simultaneously to the beta 2-appendage site and to a second site on the adjacent beta 2 hinge. This results in the displacement of the other ligands from the beta 2 appendage. Thus clathrin binding to an AP2-accessory protein complex would cause the controlled release of accessory proteins at sites of vesicle formation.

Protection of radical intermediates at the active site of adenosylcobalamin-dependent methylmalonyl-CoA mutase.

Adenosylcobalamin-dependent methylmalonyl-CoA mutase catalyzes the interconversion of methylmalonyl-CoA and succinyl-CoA via radical intermediates generated by substrate-induced homolysis of the coenzyme carbon-cobalt bond. From the structure of methylmalonyl-CoA mutase it is evident that the deeply buried active site is completely shielded from solvent with only a few polar contacts made between the protein and the substrate. Site-directed mutants of amino acid His244, a residue close to the inferred site of radical chemistry, were engineered to investigate its role in catalysis. Two mutants, His244Ala and His244Gln, were characterized using kinetic and spectroscopic techniques. These results confirmed that His244 is not an essential residue. However, compared with that of the wild type, k(cat) was lowered by 10(2)- and 10(3)-fold for the His244Gln and His244Ala mutants, respectively, while the K(m) for succinyl-CoA was essentially unchanged in both cases. The primary kinetic tritium isotope effect (k(H)/k(T)) for the His244Gln mutant was 1.5 +/- 0.3, and tritium partitioning was now found to be dependent on the substrate used to initiate the reaction, indicating that the rearrangement of the substrate radical to the product radical was extremely slow. The His244Ala mutant underwent inactivation under aerobic conditions at a rate between 1 and 10% of the initial rate of turnover. The crystal structure of the His244Ala mutant, determined at 2.6 A resolution, indicated that the mutant enzyme is unaltered except for a cavity in the active site which is occupied by an ordered water molecule. Molecular oxygen reaching this cavity may lead directly to inactivation. These results indicate that His244 assists directly in the unusual carbon skeleton rearrangement and that alterations in this residue substantially lower the protection of reactive radical intermediates during catalysis.

Dislocation and hypertrophy of the medial head of the clavicle: an unusual late complication of radical neck dissection.

We report three cases of a rare late complication of neck dissection: anterior dislocation and hypertrophy of the sternal head of the clavicle manifesting as a hard lump in the lower neck. We describe the mode of presentation, etiology, and methods of prevention.

Some notes on choices in data collection.

Collecting optimum X-ray diffraction data involves a number of choices and compromises, including choice of crystal, source, rotation range, exposure time and programs for integration and scaling. This paper presents a series of questions which should be considered in planning a data-collection experiment.

Crystal structure of substrate complexes of methylmalonyl-CoA mutase.

X-ray crystal structures of methylmalonyl-CoA mutase in complexes with substrate methylmalonyl-CoA and inhibitors 2-carboxypropyl-CoA and 3-carboxypropyl-CoA (substrate and product analogues) show that the enzyme-substrate interactions change little during the course of the rearrangement reaction, in contrast to the large conformational change on substrate binding. The substrate complex shows a 5'-deoxyadenine molecule in the active site, bound weakly and not attached to the cobalt atom of coenzyme B12, rotated and shifted from its position in the substrate-free adenosylcobalamin complex. The position of Tyralpha89 close to the substrate explains the stereochemical selectivity of the enzyme for (2R)-methylmalonyl-CoA.

A structural explanation for the binding of multiple ligands by the alpha-adaptin appendage domain.

The alpha subunit of the endocytotic AP2 adaptor complex contains a 30 kDa "appendage" domain, which is joined to the rest of the protein via a flexible linker. The 1.9 A resolution crystal structure of this domain reveals a single binding site for its ligands, which include amphiphysin, Eps15, and epsin. This domain when overexpressed in COS7 fibroblasts is shown to inhibit transferrin uptake, whereas mutants in which interactions with its binding partners are abolished do not. DPF/W motifs present in appendage domain-binding partners are shown to play a crucial role in their interactions with the domain. A single site for binding multiple ligands would allow for temporal and spatial regulation in the recruitment of components of the endocytic machinery.

Importance of the pleckstrin homology domain of dynamin in clathrin-mediated endocytosis.

The GTPase dynamin plays an essential role in clathrin-mediated endocytosis [1] [2] [3]. Substantial evidence suggests that dynamin oligomerisation around the necks of endocytosing vesicles and subsequent dynamin-catalysed GTP hydrolysis is responsible for membrane fission [4] [5]. The pleckstrin homology (PH) domain of dynamin has previously been shown to interact with phosphoinositides, but it has not been determined whether this interaction is essential for dynamin's function in endocytosis [6] [7] [8] [9]. In this study, we address the in vivo function of the PH domain of dynamin by assaying the effects of deletions and point mutations in this region on transferrin uptake in COS-7 fibroblasts. Overexpression of a dynamin construct lacking its entire PH domain potently blocked transferrin uptake, as did overexpression of a dynamin construct containing a mutation in the first variable loop of the PH domain. Structural modelling of this latter mutant suggested that the lysine residue at position 535 (Lys535) may be critical in the coordination of phosphoinositides, and indeed, the purified mutant no longer interacted with lipid nanotubes. Interestingly, the inhibitory phenotype of cells expressing this dynamin mutant was partially relieved by a second mutation in the carboxy-terminal proline-rich domain (PRD), one that prevents dynamin from binding to the Src homology 3 (SH3) domain of amphiphysin. These data demonstrate that dynamin's interaction with phosphoinositides through its PH domain is essential for endocytosis. These findings also support our hypothesis that PRD-SH3 domain interactions are important in the recruitment of dynamin to sites of endocytosis.

Fructose-sorbitol malabsorption and symptom provocation in irritable bowel syndrome: relationship to enteric hypersensitivity and dysmotility.

BACKGROUND: Fructose-sorbitol (F-S) mixtures can provoke symptoms in irritable bowel syndrome (IBS) patients, and a proportion of IBS patients also have enteric hypersensitivity to distension. We hypothesized, therefore, that sugar malabsorption and fermentation to produce hydrogen gas may provoke symptoms to a greater extent in IBS patients hypersensitive to distension than in those patients without such hypersensitivity. Our aims were therefore to compare, in IBS patients, symptoms and breath hydrogen responses after F-S, on the basis of jejunal sensitivity and jejunal motor function. METHODS: Fifteen female IBS patients (44 +/- 15 years) underwent, on separate occasions, 3-h breath hydrogen analyses after ingesting 10 g lactulose and 25 g fructose with 5 g sorbitol. Jejunal sensitivity and motor function were determined by balloon distension and 24-h manometry studies, respectively. Cumulative symptom scores and breath hydrogen production were analysed on the basis of the presence or absence of jejunal hypersensitivity and dysmotility. RESULTS: Four and seven patients had jejunal hypersensitivity for initial perception and pain, respectively. Eleven, nine, and nine patients had jejunal dysmotility for fasting phase 3, phase 2, and fed motor activity, respectively. Of the patients with symptom provocation after F-S (n = 8 within 3 h, n = 12 within 12 h) or with F-S malabsorption (n = 10), the relative proportion did not differ on the basis of the presence or absence of jejunal hypersensitivity or of motor dysfunction. Symptom scores and hydrogen production also were not different in these subgroups. CONCLUSIONS: Although carbohydrate malabsorption can provoke symptoms in some IBS patients, there is no consistent association between such a phenomenon and the presence of either jejunal hypersensitivity or dysmotility.