Ophiuroids (Echinodermata; Ophiuroidea) of biogenic habitats on the continental shelf of New Zealand.

The taxonomy of ophiuroids collected in 2009 and 2011, from biogenic habitats across the New Zealand continental shelf, is reviewed. Ophionereis novaezelandiae Mortensen, 1936, and its junior synonym Ophionereis terba Baker & Devaney, 1981 from South-Eastern Australia, is now recognised as a distinct species, and has been removed from synonymy with Ophionereis fasciata Hutton, 1872. Ophiacantha abyssicola var. otagoensis Fell, 1958 is also recognised as a distinct species and has been removed from synonymy with Ophiacantha brachygnatha Clark H L, 1928. Amphiura eugeniae var. latisquama Mortensen, 1924 is raised to species rank and Amphioplus longirima Fell, 1952 treated as a synonym of A. latisquama. Ophiolycus farquhari McKnight, 2003 is transferred to the genus Ophiologimus. The diagnostic characters of several other species are reviewed and colour descriptions and images are included where available. The tropical species Ophiacantha longidens Lyman, 1878, Ophiotreta valenciennesi (Lyman, 1879) and Ophiobyrsa intorta (Koehler, 1922) are reported from New Zealand waters for the first time.

Non-traumatic lower limb vascular complications in endurance athletes. Review of literature.

OBJECTIVES: To review the importance, clinical features, investigations, management and prognosis of non-traumatic vascular injuries, affecting the lower limbs of endurance athletes. DESIGN: Review of literature. MATERIALS AND METHODS: A literature search was conducted from Medline, Pubmed, the National Electronic Library for Health, Google and Yahoo search engines for related articles and case reports regarding non-traumatic vascular complications involving the lower limb of endurance athletes. CONCLUSIONS: Non-traumatic vascular complications affecting the lower limbs include endofibrosis, stenosis/kinking of iliac arteries, dissection of external iliac artery, adductor canal syndrome, popliteal entrapment syndrome, chronic exertional compartment syndrome and effort-induced venous thrombosis. These are important as they affect athletes at the peak of their career and can be confusing to diagnose. The management is relatively well documented and produces good results in short term but the long term results are not known.

Usefulness of immunohistochemistry in delineating renal spindle cell tumours. A retrospective study of 31 cases.

AIMS: To assess the usefulness of immunohistochemistry in delineating tumour diagnoses on a series of morphologically diagnosed renal spindle cell tumours (RSCTs). METHODS AND RESULTS: Formalin-fixed paraffin-embedded tissues from 31 morphologically diagnosed tumours were reinterpreted in light of newly obtained immunohistochemical information. By morphology, six had originally been classified as sarcomatoid carcinoma, five as spindle cell tumour (NOS), four as sarcoma (NOS), three as leiomyoma, three as leiomyosarcoma, and one each as fibrous polyp, hamartoma, neurilemmoma, mesoblastic nephroma, medullary fibroma, angiomyolipoma, haemangiopericytoma, malignant rhabdoid tumour, malignant Triton tumour, and carcinosarcoma. The application of immunohistochemistry verified the original diagnosis in 18 cases (18/31, 58%), confirming the diagnosis of sarcomatoid renal carcinoma (4/6), leiomyoma (2/3), leiomyosarcoma (3/3), sarcoma (NOS) (2/4), carcinosarcoma (1/1), malignant rhabdoid tumour (1/1), malignant Triton tumour (1/1), fibrous polyp (1/1), mesoblastic nephroma (1/1), hamartoma (1/1), and angiomyolipoma (1/1). Different tumour designations were suggested in 13 cases (13/31, 42%), including carcinosarcoma, sarcoma (NOS), leiomyosarcoma, solitary fibrous tumour, monomorphic/biphasic angiomyolipoma, endometrial stromal tumour, and congenital mesoblastic nephroma. CONCLUSIONS: Our data indicate that although morphology is most important in formulating the initial differential diagnosis, the addition of immunohistochemistry is vital in arriving at the correct classification of RSCTs.

Computer imaging and patient satisfaction in rhinoplasty surgery.

The measurement and achievement of improved patient benefit following a particular medical or surgical intervention has become an increasingly relevant part of the provision of effective healthcare. We have retrospectively analysed patient satisfaction in 56 patients following rhinoplasty via the Glasgow Benefit Inventory (GBI), 25 of whom underwent pre-operative computer imaging planning. We have also audited patient reaction to this technique via a concurrent questionnaire in those subjects who underwent imaging, and correlated this with overall patient outcome. Patient satisfaction with cosmetic rhinoplasty following computer imaging was significantly improved compared to those patients who did not receive imaging.

Synthesis, processing and export of cytoplasmic endo-beta-1,4-xylanase from barley aleurone during germination.

We have identified the major endo-beta-1,4-xylanase (XYN-1) in the aleurone of germinating barley grain, and show that it is expressed as a precursor of Mr 61 500 with both N- and C-terminal propeptides. XYN-1 is synthesized as an inactive enzyme in the cytoplasm, and only becomes active at a late stage of germination when the aleurone ceases to secrete hydrolases. A series of processing steps, mediated in part by aleurone cysteine endoproteases, yields a mature active enzyme of Mr 34 000. Processing and extracellular release of the mature enzyme coincide with the programmed cell death (PCD)-regulated disintegration of aleurone cells. We discuss the significance of delayed aleurone cell-wall degradation by endoxylanases in relation to the secretory capacity of the aleurone, and propose a novel role for aleurone PCD in facilitating the export of hydrolases.

Prediction of protein cleavage sites by the barley cysteine endoproteases EP-A and EP-B based on the kinetics of synthetic peptide hydrolysis.

Hordeins, the natural substrates of barley (Hordeum vulgare) cysteine endoproteases (EPs), were isolated as protein bodies and degraded by purified EP-B from green barley malt. Cleavage specificity was determined by synthesizing internally quenched, fluorogenic tetrapeptide substrates of the general formula 2-aminobenzoyl-P(2)-P(1)-P(1)'-P(2)' 1-tyrosine(NO(2))-aspartate. The barley EPs preferred neutral amino acids with large aliphatic and nonpolar (leucine, valine, isoleucine, and methionine) or aromatic (phenylalanine, tyrosine, and tryptophan) side chains at P(2), and showed less specificity at P(1), although asparagine, aspartate, valine, and isoleucine were particularly unfavorable. Peptides with proline at P(1) or P(1)' were extremely poor substrates. Cleavage sites with EP-A and EP-B preferred substrate sequences are found in hordeins, their natural substrates. The substrate specificity of EP-B with synthetic peptides was used successfully to predict the cleavage sites in the C-terminal extension of barley beta-amylase. When all of the primary cleavage sites in C hordein, which occur mainly in the N- and C-terminal domains, were removed by site-directed mutagenesis, the resulting protein was degraded 112 times more slowly than wild-type C hordein. We suggest that removal of the C hordein terminal domains is necessary for unfolding of the beta-reverse turn helix of the central repeat domain, which then becomes more susceptible to proteolytic attack by EP-B.

The untranslated leader sequence of the barley lipoxygenase 1 (Lox1) gene confers embryo-specific expression.

The barley lipoxygenase 1 (Lox1) gene encodes a protein expressed in embryos during grain development and germination and in leaves after methyl-jasmonate (MeJA) treatment. Transient gene expression assays in germinating barley embryos were used to identify cis-regulatory elements involved in the embryo-specific expression of the Lox1 gene. Analysis of transcriptional or translational fusions between Lox1 5' upstream sequences and the gusA reporter gene indicated that the 5'-untranslated leader sequence was involved in embryo-specific expression. Replacement of the leader sequence from the aleurone-specific Chi26 gene with the Lox1 leader sequence resulted in a chimeric gene expressed at high levels in embryo as well as in aleurone cells. Insertion of the Lox1 leader sequence between the 35S minimum promoter (A domain -90/+8) and the gusA reporter gene greatly enhanced promoter activity in a tissue-specific manner. Deletion/replacement analysis of the Lox1 leader sequence, combined with transient expression in germinating embryos and in vitro transcription/translation assays, suggests that the Lox1 leader sequence contains cis-elements regulating qualitative (tissue-specific) and quantitative gene expression.

Allele-dependent barley grain beta-amylase activity.

The wild ancestor of cultivated barley, Hordeum vulgare subsp. spontaneum (K. Koch) A. & Gr. (H. spontaneum), is a source of wide genetic diversity, including traits that are important for malting quality. A high beta-amylase trait was previously identified in H. spontaneum strains from Israel, and transferred into the backcross progeny of a cross with the domesticated barley cv Adorra. We have used Southern-blot analysis and beta-amy1 gene characterization to demonstrate that the high beta-amylase trait in the backcross line is co-inherited with the beta-amy1 gene from the H. spontaneum parent. We have analyzed the beta-amy1 gene organization in various domesticated and wild-type barley strains and identified three distinct beta-amy1 alleles. Two of these beta-amy1 alleles were present in modern barley, one of which was specifically found in good malting barley cultivars. The third allele, linked with high grain beta-amylase activity, was found only in a H. spontaneum strain from the Judean foothills in Israel. The sequences of three isolated beta-amy1 alleles are compared. The involvement of specific intron III sequences, in particular a 126-bp palindromic insertion, in the allele-dependent expression of beta-amylase activity in barley grain is proposed.

Substrate specificity of barley cysteine endoproteases EP-A and EP-B.

The cysteine endoproteases (EP)-A and EP-B were purified from green barley (Hordeum vulgare L.) malt, and their identity was confirmed by N-terminal amino acid sequencing. EP-B cleavage sites in recombinant type-C hordein were determined by N-terminal amino acid sequencing of the cleavage products, and were used to design internally quenched, fluorogenic peptide substrates. Tetrapeptide substrates of the general formula 2-aminobenzoyl-P2-P1-P1'-P2'-tyrosine(NO2)-aspartic acid, in which cleavage occurs between P1 and P1', showed that the cysteine EPs preferred phenylalanine, leucine, or valine at P2. Arginine was preferred to glutamine at P1, whereas proline at P2, P1, or P1' greatly reduced substrate kinetic specificity. Enzyme cleavage of C hordein was mainly determined by the primary sequence at the cleavage site, because elongation of substrates, based on the C hordein sequence, did not make them more suitable substrates. Site-directed mutagenesis of C hordein, in which serine or proline replaced leucine, destroyed primary cleavage sites. EP-A and EP-B were both more active than papain, mostly because of their much lower Km values.

Presencing in practice. Utilising the core of the caring process.

This article is about knowing in nursing, and using presence knowingly. It examines some descriptions from the literature and discusses how issues raised sit with my personal practice-based understanding of presencing. Four short exemplars are given to illustrate some of my experience with presencing, often in difficult circumstances, in clinical nursing practice over the years.

Identification of a methyl jasmonate-responsive region in the promoter of a lipoxygenase 1 gene expressed in barley grain.

A genomic DNA fragment was isolated containing 5' upstream sequences and part of the open reading frame corresponding to the lipoxygenase 1 cDNA (LoxA) expressed in barley grains during development and germination. Lox1 transcription was shown to be methyl jasmonate (MeJA)- and wound-inducible in leaves, but Lox1 transcripts were not detected in mildew-infected leaves, although this is a commonly observed response to pathogenic attack in various plants. Transient gene expression assays were used to identify a promoter region involved in MeJA-responsive expression. Analysis of 5' and 3' promoter deletions indicated that sequences between -363 and -294 conferred MeJA-responsive expression. Deletions/replacements covering this part of the promoter further defined a MeJA-responsive region between -331 and -291. Insertion of the region -328 to -293 into the constitutive CaMV 35S promoter conferred MeJA-responsive expression. The 36 bp fragment contains the motif TGACG as inverted repeats, which has been previously identified as a binding site for bZIP transactivating factors. Site-directed mutagenesis on these TGACG motifs abolished MeJA-responsive expression, clearly identifying them as MeJA-responsive elements. Sequence comparisons found no similar motif in other characterized promoters of MeJA-inducible genes, but suggested a common spatial structure which may serve as a binding site for transacting factors involved in the MeJA signal transduction pathway.

A synthesis of caring praxis and critical social theory in an emancipatory curriculum.

As several schools of nursing in New Zealand are accepting the challenge to prepare nurses better in order to meet changing health care needs, one lecturer discusses the design and implementation of a new degree curriculum at Manawatu Polytechnic. The author has drawn from educational, nursing and feminist literature to provide a background for the central concepts in order to explain their role in the development of the curriculum. It is contended that the caring imperative is central in all student-educator-clinician relationships when the purpose of the curriculum is to emancipate students to become nurses who care for individuals, families and communities in a transformative way. The transformative teacher is also aware of the power of open and regular dialogue when teaching caring within a critical social paradigm, and has a responsibility to ensure students understand the mandate for social action. It is through such consciousness-raising that nurses are empowered to both provide the best possible nursing care, and become agents of social change through an overhauled health care system.

Primary structure of a lipoxygenase from barley grain as deduced from its cDNA sequence.

A full length cDNA sequence for a barley grain lipoxygenase was obtained. It includes a 5' untranslated region of 69 nucleotides, an open reading frame of 2586 nucleotides encoding a protein of 862 amino acid residues and a 3' untranslated region of 142 nucleotides. The molecular mass of the encoded polypeptide was calculated to be 96.392. Its amino acid sequence shows a high homology with that of other plant lipoxygenases identified to date.

The barley 60 kDa jasmonate-induced protein (JIP60) is a novel ribosome-inactivating protein.

The N-terminal region of a 60 kDa, jasmonate-induced protein of barley leaves (JIP60) is shown to be homologous to the catalytic domains of plant ribosome-inactivating proteins (RIP). Western blotting of leaf extracts and in vitro reconstitution experiments indicate that JIP60 is synthesized as a precursor which is processed in vivo. This is in keeping with in vitro translation experiments indicating that a deletion derivative of the N-terminal region, but not the putative precursor, strongly inhibits protein synthesis on reticulocyte ribosomes. The inhibition of ribosome function is associated with depurination of 26S rRNA, characteristic of plant RIPs. This indicates that JIP60 is a novel ribosome-inactivating protein requiring at least two processing events for full activation. JIP60 derivatives do not significantly inhibit in vitro protein synthesis on wheat germ ribosomes. These and other results suggest that JIP60 may be involved in plant defence.

The expression of serine carboxypeptidases during maturation and germination of the barley grain.

cDNA clones encoding three additional serine carboxypeptidases (Ser-CPs) have been isolated from a gibberellic acid-induced barley aleurone cDNA library. The three deduced Ser-CPs belong to the two-chain subfamily of Ser-CPs; they are synthesized as precursors with a putative signal peptide, propeptide, and linker peptide between the A and B chains. Their identification provides the proof for the existence of more than three Ser-CPs in cereal grains, and, based on their sequences, they may exhibit new substrate specificities. The expression of these and of the three previously isolated Ser-CPs from barley grains (CP-MI, CP-MII, and CP-MIII) has been investigated by Northern and Western analysis and RNA PCR. CP-MII is the only Ser-CP to be expressed and accumulate in the developing grain and is stored in its active form in the mature grain. All six Ser-CPs are expressed de novo in the germinating grain, in the scutellum, and/or in the aleurone. Furthermore, at least CP-MI, CP-MII, and CP-MIII are secreted into the endosperm. In addition, all Ser-CPs (except CP-MI) are also expressed in the roots and shoots of the growing seedling. This enzyme family thus appears to be ubiquitous in the barley plant, which suggests that Ser-CPs play additional roles besides their participation in the mobilization of storage proteins.

A role for gamma 3 hordein in the transport and targeting of prolamin polypeptides to the vacuole of developing barley endosperm.

Hordein synthesis, transport and deposition was analysed by immunocytochemistry in developing endosperm cells of wild-type (Carlsberg II) and mutant varieties deficient in B hordein (hor2ca), gamma 1 hordein (Donetsky), gamma 2 hordein and minor B hordein polypeptides (Haisa), or gamma 3 hordein (Nevsky). In all varieties, hordein polypeptides were detected both in the cytoplasm as globules, ranging in diameter from 50 nm to 1.24 microns, and in the vacuole as protein bodies. In the cytoplasmic globules B and C hordein polypeptides are assembled as a core and are surrounded by an outer layer of gamma 1 and gamma 2 hordein. The globules apparently fuse several times in the cytoplasm before entering the vacuole. Absence of gamma 3 hordein in the mutant Nevsky leads to a dramatic change in hordein polypeptide targeting, the hordein storage proteins being largely deposited in the lumen of the rough endoplasmic reticulum. gamma 3 Hordein is unique among the sulphur-rich hordein polypeptides, being monomeric and forming only intramolecular disulphide bridges, while the other B and gamma hordein polypeptides are aggregated by intermolecular disulphide bridges. Retention of hordein in the rough endoplasmatic reticulum in the absence of gamma 3 hordein suggests that gamma 3 hordein may maintain the prolamin storage polypeptides in a transport competent state. The sequence of the mature gamma 3 hordein polypeptide was deduced from a cDNA clone, and compared with gamma 2 hordein. The epitope recognized by the gamma 1 + gamma 2 hordein-specific BX monoclonal antibody used for immunocytochemistry was mapped to include E190 and K193, by synthesizing overlapping oligopeptides.

Evolutionary relationship of the members of the sulphur-rich hordein family revealed by common antigenic determinants.

Five monoclonal antibodies raised against an enriched C hordein fraction have been characterized in detail and were found to be specific for the members of the sulphur-rich hordein family. Two antibodies specific for B hordein polypeptides were identified, one of which reacted predominantly with CNBr cleavage class III polypeptides. γ1 hordein was recognized by two antibodies, of which one also reacted with γ2 hordein and several members of the CNBr cleavage class II B hordein polypeptides. One antibody recognized γ3 hordein but cross-reacted at higher antibody concentration with almost all of the B and C hordein polypeptides. The specificity of the monoclonal antibodies was confirmed by Western blotting of one- or two-dimensionally separated hordein from the B hordein-deficient mutant hor2ca and its wild-type Carlsberg II and the γ3 hordein-deficient genotype Nevsky. The identification of the γ hordein-specific monoclonal antibodies was further supported by immune precipitation of in-vitro transcribed and translated γ2 hordein, and hor2ca and Carlsberg II mRNA translation products. The monoclonal antibodies were used to screen for mutants in γ hordein synthesis. Two mutants, one deficient in γ 1 hordein synthesis and a second in γ 2 or closely related B hordein polypeptides were identified. A model is proposed for the evolution of the sulphur-rich hordein loci Hor5 and Hor2.