Journal articles on the topic 'Bruce (donor)'

To prevent the wooden door frame of traditional rural houses from being stuck due to diamond deformation under earthquake and improve the seismic capacity of rural houses, an innovative method of reinforcing the angular displacement of the wooden door frame with channel steel and the diagonal brace is proposed. The rationality of the finite element simulation is demonstrated by comparing the results of finite element simulation and quasistatic test based on reinforced and unreinforced wooden door frame specimens. On the basis of the finite element model of wooden door frame, the seismic performance of channel type and diagonal brace thickness of reinforced wooden door frame and the seismic performance of friction coefficient of unreinforced wooden door frame are studied, respectively. The results show that the lateral stiffness and the lateral bearing capacity of the reinforced wooden door frame increase with the increase of channel steel type and the diagonal brace thickness. The height of the channel steel section of the seismic reinforcement structure should be half of the unreinforced structure. With the increase of the friction coefficient, the lateral bearing capacity of the unreinforced wooden frame increases, while the ductility of the unreinforced wooden frame decreases.

Trypanosoma brucei brucei and other trypanosomatid species are unique among eucaryotes because transcription of their protein-coding genes is discontinuous. The 5' ends of their mRNAs consist of an identical 35-nucleotide spliced leader which is encoded at a separate locus from that for the body of the protein-coding transcript. We show here that the spliced leader transcript contains a 5' cap structure and suggest that at least one function of the spliced leader sequence is to provide a cap structure to trypanosome mRNAs.

Trypanosoma brucei brucei and other trypanosomatid species are unique among eucaryotes because transcription of their protein-coding genes is discontinuous. The 5' ends of their mRNAs consist of an identical 35-nucleotide spliced leader which is encoded at a separate locus from that for the body of the protein-coding transcript. We show here that the spliced leader transcript contains a 5' cap structure and suggest that at least one function of the spliced leader sequence is to provide a cap structure to trypanosome mRNAs.

Exposure of recently mated female rodents to unfamiliar male scents during daily prolactin surges results in pregnancy failure (the ‘Bruce effect’). Control of nasal contact with male scents during these narrow windows of sensitivity could allow females to maintain or terminate pregnancy, but female behavioural changes specifically during this critical period have not been investigated. We examined the approach or avoidance of familiar stud strain and unfamiliar male scents by recently mated female mice. Females that maintained pregnancy avoided both unfamiliar and familiar male scent during critical periods of susceptibility for the Bruce effect. By contrast, females that did not maintain pregnancy showed a sharp rise in the time spent with unfamiliar male scent during this critical period. Manipulation of the social status of unfamiliar and stud strain scent donors did not affect the likelihood of pregnancy block, although females spent more time with dominant male scents across all time periods. The ability to control the Bruce effect through behaviour during brief sensitivity just before dusk, when females are likely to be in nest sites, provides a mechanism by which females may adjust their reproductive investment according to nest site social stability and likelihood of offspring survival.

Here we report the presence of a trans-sialidase on the surface of Trypanosoma brucei culture-derived procyclic trypomastigotes. The enzyme is not detected in lysates of bloodstream trypomastigotes enriched for either stumpy or slender forms. The trans-sialidase catalyzes the transfer of alpha(2-3)-linked sialic acid residues to lactose. beta-galactopyranosyl residues are at least 100 times better acceptors for sialic acid than alpha-galactopyranosyl residues. In the absence of efficient acceptors, the purified enzyme transfers sialic acid to water, i.e., it acts as a sialidase. Although the T. cruzi and T. brucei trans-sialidases have very similar donor and acceptor specificities, they are antigenically distinct. Sodium dodecyl sulfate-polyacramide gel electrophoresis under nonreducing conditions and silver staining of the purified trans-sialidase reveals a single band of 63 kD. When the surface membrane of live procyclic trypomastigotes is trans-sialylated, using radioactive sialyllactose as the donor substrate, it appears that the only sialylated surface molecule is procyclin. Pronase treatment of live parasites removes only part of the surface sialic acid, in agreement with recent data showing that the glycosylphosphatidylinositol anchor of procyclin is sialylated (Ferguson, M. A. J., M. Murray, H. Rutherford, and M. J. McConville. 1993. Biochem. J. In press).

ABSTRACTNitroheterocyclic prodrugs are used to treat infections caused byTrypanosoma cruziandTrypanosoma brucei. A key component in selectivity involves a specific activation step mediated by a protein homologous with type I nitroreductases, enzymes found predominantly in prokaryotes. Using data from determinations based on flavin cofactor, oxygen-insensitive activity, substrate range, and inhibition profiles, we demonstrate that NTRs fromT. cruziandT. bruceidisplay many characteristics of their bacterial counterparts. Intriguingly, both enzymes preferentially use NADH and quinones as the electron donor and acceptor, respectively, suggesting that they may function as NADH:ubiquinone oxidoreductases in the parasite mitochondrion. We exploited this preference to determine the trypanocidal activity of a library of aziridinyl benzoquinones against bloodstream-formT. brucei. Biochemical screens using recombinant NTR demonstrated that several quinones were effective substrates for the parasite enzyme, havingKcat/Kmvalues 2 orders of magnitude greater than those of nifurtimox and benznidazole. In tests againstT. brucei, antiparasitic activity mirrored the biochemical data, with the most potent compounds generally being preferred enzyme substrates. Trypanocidal activity was shown to be NTR dependent, as parasites with elevated levels of this enzyme were hypersensitive to the aziridinyl agent. By unraveling the biochemical characteristics exhibited by the trypanosomal NTRs, we have shown that quinone-based compounds represent a class of trypanocidal compound.

The process of trans splicing is essential to the maturation of all mRNAs in the Trypanosomatidae, a family of protozoan parasites, and to specific mRNAs in several species of nematode. In Trypanosoma brucei, a 39-nucleotide (nt) leader sequence originating from a small, 139-nt donor RNA (the spliced leader [SL] RNA) is spliced to the 5' end of mRNAs. An intermediate in this trans-splicing process is a Y structure which contains the 3' 100 nt of the SL RNA covalently linked to the pre-mRNA via a 2'-5' phosphodiester bond at the branch point residue. We mapped the branch points in T. brucei alpha- and beta-tubulin pre-mRNAs. The primary branch acceptors for the alpha- and beta-tubulins are 44 and 56 nt upstream of the 3' splice sites, respectively, and are A residues. Minor branch acceptors were detected 42 and 49 nt upstream of the alpha-tubulin splice site and 58 nt upstream of the splice site in beta-tubulin. The regions surrounding these branch points lack homology to the consensus sequences determined for mammalian cells and yeasts; there is also no conservation among the sequences themselves. Thus, the identified sequences suggest that the mechanism of branch point recognition in T. brucei differs from the mechanism of recognition by U2 RNA that has been proposed for other eucaryotes.

The process of trans splicing is essential to the maturation of all mRNAs in the Trypanosomatidae, a family of protozoan parasites, and to specific mRNAs in several species of nematode. In Trypanosoma brucei, a 39-nucleotide (nt) leader sequence originating from a small, 139-nt donor RNA (the spliced leader [SL] RNA) is spliced to the 5' end of mRNAs. An intermediate in this trans-splicing process is a Y structure which contains the 3' 100 nt of the SL RNA covalently linked to the pre-mRNA via a 2'-5' phosphodiester bond at the branch point residue. We mapped the branch points in T. brucei alpha- and beta-tubulin pre-mRNAs. The primary branch acceptors for the alpha- and beta-tubulins are 44 and 56 nt upstream of the 3' splice sites, respectively, and are A residues. Minor branch acceptors were detected 42 and 49 nt upstream of the alpha-tubulin splice site and 58 nt upstream of the splice site in beta-tubulin. The regions surrounding these branch points lack homology to the consensus sequences determined for mammalian cells and yeasts; there is also no conservation among the sequences themselves. Thus, the identified sequences suggest that the mechanism of branch point recognition in T. brucei differs from the mechanism of recognition by U2 RNA that has been proposed for other eucaryotes.

The eukaryotic enzyme NMT (myristoyl-CoA:protein N-myristoyltransferase) has been characterized in a range of species from Saccharomyces cerevisiae to Homo sapiens. NMT is essential for viability in a number of human pathogens, including the fungi Candida albicans and Cryptococcus neoformans, and the parasitic protozoa Leishmania major and Trypanosoma brucei. We have purified the Leishmania and T. brucei NMTs as active recombinant proteins and carried out kinetic analyses with their essential fatty acid donor, myristoyl-CoA and specific peptide substrates. A number of inhibitory compounds that target NMT in fungal species have been tested against the parasite enzymes in vitro and against live parasites in vivo. Two of these compounds inhibit TbNMT with IC50 values of <1 μM and are also active against mammalian parasite stages, with ED50 (the effective dose that allows 50% cell growth) values of 16–66 μM and low toxicity to murine macrophages. These results suggest that targeting NMT could be a valid approach for the development of chemotherapeutic agents against infectious diseases including African sleeping sickness and Nagana.

SUMMARYAutophosphorylation by protein kinases has been implicated as an important control mechanism in signal transduction and growth regulatory pathways in mammalian cells. We have set out to investigate whether any such autophosphorylating protein kinase activities can be found in Trypanosoma brucei. In order to do this, we have developed a system for characterizing such protein kinase activities using an in vitro assay. This assay was carried out by fractionation of trypanosome lysates using isoelectric focusing gel electrophoresis followed by incubation of the gel in γ32P-labelled nucleotide triphosphate and subsequent autoradiography. We have identified two classes of autophosphorylating protein kinase activities. In the first class all were dependent on ATP as the phosphate donor substrate and were all found to have a molecular size of 60 kDa. Differences in the activity of these protein kinases were observed between the bloodstream and procyclic life-cyle stages. Furthermore, the addition of mammalian epidermal growth factor to bloodstream stage lysates stimulated an additional activity. The second class of autophosphorylating protein kinases utilized GTP as the phosphate donor and were all found to be 90 kDa in size. Stage-specific differences were also observed in the activity of these protein kinases.

Attached type lifting scaffold is an important engineering machine which is used for building construction. In this paper, the strength of door-type structure of attached type lifting scaffold is studied. The diagonal brace is removed in the new structure and replaced with two short braces. The two type structure elastic FE models are founded and ANSYS software is used to compute the stress and deformation. The results are compared. The comparison study shows the new structure’ strength and stiffness are both better than the old one. Therefore, the new structure of door-type frame is much better than the old structure not only in cost-saving but also in easy through.

The trypanosomatids are generally aberrant in their protein N-glycosylation pathways. However, protein N-glycosylation in the African trypanosome Trypanosoma brucei, etiological agent of human African sleeping sickness, is not well understood. Here, we describe the creation of a bloodstream-form T. brucei mutant that is deficient in the endoplasmic reticulum enzyme glucosidase II. Characterization of the variant surface glycoprotein, the main glycoprotein synthesized by the parasite with two N-glycosylation sites, revealed unexpected changes in the N-glycosylation of this molecule. Structural characterization by mass spectrometry, nuclear magnetic resonance spectroscopy, and chemical and enzymatic treatments revealed that one of the two glycosylation sites was occupied by conventional oligomannose structures, whereas the other accumulated unusual structures in the form of Glcα1–3Manα1–2Manα1–2Manα1–3(Manα1–6)Manβ1–4GlcNAcβ1–4GlcNAc, Glcα1–3Manα1–2Manα1–2Manα1–3(GlcNAcβ1–2Manα1–6)Manβ1–4GlcNAcβ1–4GlcNAc, and Glcα1–3Manα1–2Manα1–2Manα1–3(Galβ1–4GlcNAcβ1–2Manα1–6)Manβ1–4GlcNAcβ1–4GlcNAc. The possibility that these structures might arise from Glc1Man9GlcNAc2 by unusually rapid α-mannosidase processing was ruled out using a mixture of α-mannosidase inhibitors. The results suggest that bloodstream-form T. brucei can transfer both Man9GlcNAc2 and Man5GlcNAc2 to the variant surface glycoprotein in a site-specific manner and that, unlike organisms that transfer exclusively Glc3Man9GlcNAc2, the T. brucei UDP-Glc: glycoprotein glucosyltransferase and glucosidase II enzymes can use Man5GlcNAc2 and Glc1Man5GlcNAc2, respectively, as their substrates. The ability to transfer Man5GlcNAc2 structures to N-glycosylation sites destined to become Man4–3GlcNAc2 or complex structures may have evolved as a mechanism to conserve dolichol-phosphate-mannose donors for glycosylphosphatidylinositol anchor biosynthesis and points to fundamental differences in the specificities of host and parasite glycosyltransferases that initiate the synthesis of complex N-glycans.

Myristate is the exclusive fatty acid species in the glycosyl phosphatidylinositol (GPI) anchor of the Trypanosoma brucei variant surface glycoprotein (VSG). [3H]Myristate can be incorporated into T. brucei GPIs by two distinct processes known as fatty acid remodelling and myristate exchange. Myristoyllysophosphatidylcholine (M-LPC) can also serve as a myristate donor for VSG in trypanosomes [Bowes, Samad, Jiang, Weaver and Mellors (1993) J. Biol. Chem. 268, 13885–13892]. We have studied in detail the myristoylation of GPIs using a [3H]M-LPC substrate. Labelling of VSG and free GPIs by [3H]M-LPC in cultured trypanosomes occurred at the same rate as with [3H]myristate. Concurrent with GPI labelling, there was rapid hydrolysis of [3H]M-LPC to generate extracellular [3H]myristate. Experiments in a trypanosomal cell-free system indicated that GPI labelling by fatty acid remodelling and myristate exchange was also equally efficient with [3H]M-LPC and [3H]myristate. Furthermore, both ATP and CoA are required for the myristoylation of GPIs by [3H]M-LPC. These experiments suggest that GPI myristoylation from M-LPC involves hydrolysis of M-LPC to free myristate. To address the physiological importance of myristate and M-LPC in VSG myristoylation, we radiolabelled trypanosomes in vivo with both substrates in medium containing serum, and found that [3H]myristate labelled VSG and GPIs more efficiently. Thus, VSG myristoylation by free myristate may be favoured in bloodstream trypanosome infections.

TbTDPX (Trypanosoma brucei tryparedoxin-dependent peroxidase) is a genetically validated drug target in the fight against African sleeping sickness. Despite its similarity to members of the GPX (glutathione peroxidase) family, TbTDPX2 is functional as a monomer, lacks a selenocysteine residue and relies instead on peroxidatic and resolving cysteine residues for catalysis and uses tryparedoxin rather than glutathione as electron donor. Kinetic studies indicate a saturable Ping Pong mechanism, unlike selenium-dependent GPXs, which display infinite Km and Vmax values. The structure of the reduced enzyme at 2.1 Å (0.21 nm) resolution reveals that the catalytic thiol groups are widely separated [19 Å (0.19 nm)] and thus unable to form a disulphide bond without a large conformational change in the secondary-structure architecture, as reported for certain plant GPXs. A model of the oxidized enzyme structure is presented and the implications for small-molecule inhibition are discussed.

Davi Kopenawa e Bruce Albert realizam juntos A queda do céu: palavras de um xamã yanomami. Considerado uma profecia, este livro de autoria complexa dá conta de um mundo (não só yanomami) visitado, colonizado, catequizado, invadido, defendido, ameaçado. O outro, aí, é o napë, palavra que passa a se referir ao conceito de branco, além de inimigo e estrangeiro. A partir da relação entre Kopenawa e Albert, proponho ler A queda do céu como performance que visa uma hospitalidade ainda desconhecida, que dissolveria o paradoxo mesmo que a constitui: pois aquele que é hospitaleiro deve ser o dono da casa, o proprietário, aquele que tem direito a um território e que, assim, o abriria ao outro. Minha hipótese é de que a hospitalidade por vir de Jacques Derrida seja possível hoje para um Yanomami e na relação que ele impõe a quem acorda, aos que dizem um tipo de “sim” no ato de ler A queda do céu. Ademais, A queda do céu, com sua origem múltipla, humana e não humana, corrobora uma autoria que desmente a propriedade e que impele o leitor a uma tomada de posição.

ABSTRACT In kinetoplastids spliced leader (SL) RNA is trans-spliced onto the 5′ ends of all nuclear mRNAs, providing a universal exon with a unique cap. Mature SL contains an m7G cap, ribose 2′-O methylations on the first four nucleotides, and base methylations on nucleotides 1 and 4 (AACU). This structure is referred to as cap 4. Mutagenized SL RNAs that exhibit reduced cap 4 are trans-spliced, but these mRNAs do not associate with polysomes, suggesting a direct role in translation for cap 4, the primary SL sequence, or both. To separate SL RNA sequence alterations from cap 4 maturation, we have examined two ribose 2′-O-methyltransferases in Trypanosoma brucei. Both enzymes fall into the Rossmann fold class of methyltransferases and model into a conserved structure based on vaccinia virus homolog VP39. Knockdown of the methyltransferases individually or in combination did not affect growth rates and suggests a temporal placement in the cap 4 formation cascade: TbMT417 modifies A2 and is not required for subsequent steps; TbMT511 methylates C3, without which U4 methylations are reduced. Incomplete cap 4 maturation was reflected in substrate SL and mRNA populations. Recombinant methyltransferases bind to a methyl donor and show preference for m7G-capped RNAs in vitro. Both enzymes reside in the nucleoplasm. Based on the cap phenotype of substrate SL stranded in the cytosol, A2, C3, and U4 methylations are added after nuclear reimport of Sm protein-complexed substrate SL RNA. As mature cap 4 is dispensable for translation, cap 1 modifications and/or SL sequences are implicated in ribosomal interaction.

AbstractPatellar bone–tendon–bone (pBTB) autografts are often considered the “gold standard” for complete anterior cruciate ligament (ACL) reconstruction and are also associated with significant complications and early-onset knee osteoarthritis (OA). A novel quadriceps tendon allograft with synthetic augmentation, or “internal brace” (QTIB), has been reported to have potential advantages for ACL reconstruction based on animal model data. In this preclinical canine comparison study, we hypothesized that QTIB allograft compared with pBTB autograft would provide superior durability for knee stability, function, and prevention of OA. Under approval from our Institutional Animal Care and Use Committee, adult purpose-bred research hounds (n = 10) underwent arthroscopic complete transection of the ACL followed by either an arthroscopic-assisted all-inside ACL reconstruction using the QTIB allograft (n = 5) or pBTB autograft (n = 5). Contralateral knees were used as nonoperated controls (n = 10). Radiographic and arthroscopic assessments were performed at 2 and 6 months, respectively, after surgery. Anterior drawer, internal rotation, lameness, kinetics, pain, effusion, and comfortable range of knee motion were measured at 2, 3, and 6 months. Biomechanical and histologic assessments were performed at 6 months. All reconstructed knees were stable and had intact ACL grafts 6 months after surgery. At 6 months, QTIB reconstructed knees had significantly less lameness, lower pain, less effusion, and increased range of motion when compared with BTB knees (p < 0.05). BTB knees had significantly higher radiographic OA scores than QTIB knees at 6 months (p < 0.05). Superior outcomes associated with QTIB allograft may be due to the lack of donor site morbidity, the use of a robust tendon graft, and/or protection of the graft from the synthetic augmentation. Robust tendon grafts combined with a synthetic internal brace and platelet-rich plasma (PRP) may allow for more rapid and robust tendon–bone healing and graft “ligamentization,” which protects the graft from early failure and rapid OA development that can plague commonly-used allografts.

Since Robert Dahl's seminal writings on democracy more than two decades ago, interest in the topic has emerged again, especially among scholars analyzing democratic transitions. Great strides have been made in revealing the uncertain nature of these transitions (O'Donnell et al. 1986; Malloy and Seligson 1987; Diamond, Linz, and Lipset 1989; Hakim and Lowenthal 1991; O'Donnell 1994), in methodologically analyzing them as contested and “crafted” rather than spontaneous (Di Palma 1990), and in documenting the class and social forces that make democratic outcomes more likely (Rueschemeyer, Stephens, and Stephens 1992; see also Moore 1966). Despite these advances, there has been little change in our theoretical understanding of democracy. As Bruce Cumings has perceptively noted, recent studies of democratic transition have “given way to atheoretical and idiosyncratic explanations of more or less successful democratic ‘openings’” in which little time is spent elaborating “the decision rule for saying this person is hard-line or soft-line, that system is ‘liberalized autocracy’ instead of ‘limited democracy,’” or for defining democracy itself. If scholars do bring theory into their writings “through the back door of the obscure but telling footnote,” he observes, “rather than advancing their own conception of democracy, [they] uniformly define democracy by reference to Robert Dahl's Polyarchy, a classic pluralist account of the North American system” (Cumings 1989:15–17).

ABSTRACT Aspartate aminotransferases have been cloned and expressed fromCrithidia fasciculata, Trypanosoma brucei brucei, Giardia intestinalis, andPlasmodium falciparum and have been found to play a role in the final step of methionine regeneration from methylthioadenosine. All five enzymes contain sequence motifs consistent with membership in the Ia subfamily of aminotransferases; the crithidial and giardial enzymes and one trypanosomal enzyme were identified as cytoplasmic aspartate aminotransferases, and the second trypanosomal enzyme was identified as a mitochondrial aspartate aminotransferase. The plasmodial enzyme contained unique sequence substitutions and appears to be highly divergent from the existing members of the Ia subfamily. In addition, the P. falciparum enzyme is the first aminotransferase found to lack the invariant residue G197 (P. K. Mehta, T. I. Hale, and P. Christen, Eur. J. Biochem. 214:549–561, 1993), a feature shared by sequences discovered in P. vivax and P. berghei. All five enzymes were able to catalyze aspartate-ketoglutarate, tyrosine-ketoglutarate, and amino acid-ketomethiobutyrate aminotransfer reactions. In the latter, glutamate, phenylalanine, tyrosine, tryptophan, and histidine were all found to be effective amino donors. The crithidial and trypanosomal cytosolic aminotransferases were also able to catalyze alanine-ketoglutarate and glutamine-ketoglutarate aminotransfer reactions and, in common with the giardial aminotransferase, were able to catalyze the leucine-ketomethiobutyrate aminotransfer reaction. In all cases, the kinetic constants were broadly similar, with the exception of that of the plasmodial enzyme, which catalyzed the transamination of ketomethiobutyrate significantly more slowly than aspartate-ketoglutarate aminotransfer. This result obtained with the recombinant P. falciparum aminotransferase parallels the results seen for total ketomethiobutyrate transamination in malarial homogenates; activity in the latter was much lower than that in homogenates from other organisms. Total ketomethiobutyrate transamination in Trichomonas vaginalis and G. intestinalis homogenates was extensive and involved lysine-ketomethiobutyrate enzyme activity in addition to the aspartate aminotransferase activity. The methionine production in these two species could be inhibited by the amino-oxy compounds canaline and carboxymethoxylamine. Canaline was also found to be an uncompetitive inhibitor of the plasmodial aspartate aminotransferase, with aKi of 27 μM.

La presente investigación cualitativa, de abordaje metodológico estudio de caso, tuvo como objetivo describir, analizar y comprender la percepción de las amas de casa sobre la postura corporal al usar la cocina mejorada, Inkawasi - Lambayeque 2015. El referencial teórico sustentado por Bruce (2006) sobre la percepción, Bonito (2004) fundamenta las posturas corporales y Barrick (2011) respalda la importancia de las cocinas mejoradas. Los sujetos de investigación fueron 6 mujeres dedicadas a las tareas del hogar, obtenidas por método de saturación Se realizó la entrevista semiestructurada, y el análisis de contenido temático, emergiendo dos categorías: (1) Percepción de las amas de casa sobre la postura corporal antes de usar la cocina mejorada; (2) Percepción de los cambios posturales que muestran las amas de casa al utilizar las cocinas mejoradas, se tuvo en cuenta los criterios de rigor científico de Morse y los de rigor ético según Sgreccia. Se concluye, que las mujeres de Inkawasi percibieron que la inadecuada postura corporal por el uso de las cocinas tradicionales, les produjo efectos negativos en su salud, y al implementarse la cocina mejorada, les ha generado grandes beneficios, como la disminución del esfuerzo al manipular carga, alivio del dolor en la zona de la espalda, rodillas, brazos y cintura, Además la altura de la cocina les ha permitido deslizar las ollas horizontalmente con mayor eficacia reduciendo la fatiga y el riesgo de lesiones músculo – esqueléticos. Por otro lado al adoptar los cambios posturales, las amas de casa de Inkawasi, se han sentido cómodas, tranquilas, con un mayor confort, mejorando su salud y por ende su calidad de vida.

The purpose of this study was to compare the psychomotor performance of students taught woodwork technology with classroom as against online mode of instruction in Nigerian Universities. Two research questions guided the study, as well, one (1) hypothesis was formulated and tested at .05 level of significance. The quasi experimental design was used and the population of the study was 67Undergraduate Students from the Department of Technical Education, Ebonyi State University Abakiliki. The sample of the study was 48 students comprising of 30 and 18 students from 100level and 200level respectively. The instrument for data collection was the Woodwork Technology Practical Skills Assessment Instrument (WTPSAI) for measuring the psycho-performance of Woodwork Technology students in the manufacturing of stool, office chairs, door frame, panel door, and brace and batten door. The instrument was developed by the researcher which was content and face validated by 3 research experts from Delta State University Abraka, one from Measurement and Evaluation and the other two from Technical Education. Treatment was administered to the two experimental groups using two different mode of instruction, after which data were collected by the researcher through the administration of WTPSAI by lecturer from the institution selected for the study. Data collected were analysed using T-test to test the hypothesis. The study revealed that classroom and online mode of instruction for teaching and learning woodwork technology have significant effects on students’ psychomotor performance in Woodwork Technology. Furthermore, unlike Classroom mode of instruction, Online Mode of Instruction appears to be more effective in terms of the acquisition of psychomotor skills. Based on the findings of the study, it was recommended among others that Woodwork Technology lecturers and instructors should integrate online mode of instruction in the implementation of Woodwork Technology curriculum which will aid in the production of competent woodwork Technology graduates that will be useful to him/her self and to the society.

Retrotransposon Hot Spot (RHS) is the most abundant gene family in Trypanosoma cruzi, with unknown function in this parasite. The aim of this work was to shed light on the organization and expression of RHS in T. cruzi. The diversity of the RHS protein family in T. cruzi was demonstrated by phylogenetic and recombination analyses. Transcribed sequences carrying the RHS domain were classified into ten distinct groups of monophyletic origin. We identified numerous recombination events among the RHS and traced the origins of the donors and target sequences. The transcribed RHS genes have a mosaic structure that may contain fragments of different RHS inserted in the target sequence. About 30% of RHS sequences are located in the subtelomere, a region very susceptible to recombination. The evolution of the RHS family has been marked by many events, including gene duplication by unequal mitotic crossing-over, homologous, as well as ectopic recombination, and gene conversion. The expression of RHS was analyzed by immunofluorescence and immunoblotting using anti-RHS antibodies. RHS proteins are evenly distributed in the nuclear region of T. cruzi replicative forms (amastigote and epimastigote), suggesting that they could be involved in the control of the chromatin structure and gene expression, as has been proposed for T. brucei.

The crude aqueous extract of Guiera senegalensis stem bark was evaluated for its phytochemistry, acute toxicity and in vitro antitrypanosomal efficacy in this study. Tests for alkaloids, flavonoids, tannins, phlabotannins, saponins, steroids, cardenolides, terpenoids, cardiac glycosides, and anthraquinones were conducted. A total of 15 albino rats of both sexes were used and grouped into 5 (A to E) of 3 rats each. Groups A-D were intraperitoneally treated with graded doses of 100, 200, 400, 800mg/kg body weight of the crude aqueous extract of G. senegalensis stem bark. Group E was treated with Physiological Saline Solution serving as the control. All groups were observed for 24 hours for clinical signs and death to determine the median lethal dose (LD50). An in vitro experiment was carried out with 2 drops of blood from a donor rat added to 5 ml of phosphate buffer glucose solution out of which 0.2ml was finally used at 40, 20, 10, 5, 2.5, 0.625, 0.313, 0.156 and 0.078 concentrations of the extract. The phytochemical screening for bioactive substances had tannins, terpenoids, alkaloids, flavonoids, saponins, anthraquinones and cardiac glycosides. Phlabotannins and cardenolides were not detected. The clinical signs observed were sluggishness, awkward posture, loss of appetite, starry hair coat and terminal death within 24 hours with LD50 value of 600mg/kg. The in vitro antitrypanosomal efficacy of the extract showed 100% inhibition of motility against Trypanosoma brucei at 20mg/ml. In conclusion, the crude aqueous extract of G. senegalensis stem bark contains phytochemical components that exhibit inhibitory trypanosomal activity.

Abstract OBJECTIVE: Rheumatoid arthritis frequently affects the craniovertebral junction (CVJ) and may lead to severe neck pain, quadriparesis, and respiratory dysfunction. Surgery in rheumatoid nonambulatory (Ranawat Class IIIb) patients carries a significant risk. This study presents the surgical outcome of Class IIIb patients with CVJ rheumatoid myelopathy and reviews the literature. METHODS: One hundred twelve consecutive patients with rheumatoid cervical myelopathy underwent surgical decompression and stabilization. Thirty-two of the patients (mean age, 66.81 ± 10.25 yr) with CVJ rheumatoid arthritis were in Class IIIb, and all had atlantoaxial subluxation. A halo brace was applied before surgery and continued during surgery. Eleven patients with reducible atlantoaxial subluxation underwent direct posterior fusion. Twenty-one patients with fixed atlantoaxial subluxation underwent transoral decompression and then posterior fusion while they were under anesthesia. RESULTS: At a mean follow-up of 39 months, four patients improved to Class II and 14 improved to Class IIIa, whereas six remained in Class IIIb. Neck pain was relieved in all patients. There was one perioperative death after transoral surgery (posterior fusion not done), and seven other patients died subsequently of causes unrelated to surgery. The morbidity of surgery included construct failure, cerebrospinal fluid leak, superficial wound or graft donor site infection, transient dysphagia, and lung infection. CONCLUSION: A large subset of patients with CVJ rheumatoid myelopathy may reach Class IIIb. These patients have unique management considerations. Surgery (despite high morbidity) often remains the best therapeutic option available to them. Improvement of even one grade in their Ranawat score from Class IIIb to Class IIIa brought about by surgery confers on them a significant benefit in terms of their quality of life and survival.

ABSTRACT It has been shown that the coexistence of methanogenesis and reductive acetogenesis in the hindgut of the wood-feeding termiteReticulitermes flavipes is based largely on the radial distribution of the respective microbial populations and relatively high hydrogen partial pressures in the gut lumen. Using Clark-type microelectrodes, we showed that the situation in Cubitermes orthognathus and other soil-feeding members of the subfamily Termitinae is different and much more complex. All major compartments of agarose-embedded hindguts were anoxic at the gut center, and high H2 partial pressures (1 to 10 kPa) in the alkaline anterior region rendered the mixed segment and the third proctodeal segment (P3) significant sources of H2. Posterior to the P3 segment, however, H2 concentrations were generally below the detection limit (<100 Pa). All hindgut compartments turned into efficient hydrogen sinks when external H2 was supplied, but methane was formed mainly in the P3/4a and P4b compartments, and in the latter only when H2 or formate was added. Addition of H2 to the gas headspace stimulated CH4 emission of living termites, indicating that endogenous H2production limits methanogenesis also in vivo. At the low H2 partial pressures in the posterior hindgut, methanogens would most likely outcompete homoacetogens for this electron donor. This might explain the apparent predominance of methanogenesis over reductive acetogenesis in the hindgut of soil-feeding termites, although the presence of homoacetogens in the anterior, highly alkaline region cannot yet be excluded. In addition, the direct contact of anterior and posterior hindgut compartments in situ permits a cross-epithelial transfer of H2 or formate, which would not only fuel methanogenesis in these compartments, but would also create favorable microniches for reductive acetogenesis. In situ rates and spatial distribution of H2-dependent acetogenic activities are addressed in a companion paper (A. Tholen and A. Brune, Appl. Environ. Microbiol. 65:4497–4505, 1999).

In recent years, the number of reported Human African Trypanosomiasis (HAT) cases caused by Trypanosoma brucei (T.b.) gambiense has been markedly declining, and the goal of ‘elimination as a public health problem’ is within reach. For the next stage, i.e. interruption of HAT transmission by 2030, intensive screening and surveillance will need to be maintained, but with tools and strategies more efficiently tailored to the very low prevalence. We assessed the sequential use of ELISA and Immune Trypanolysis (ITL) on dried blood spot (DBS) samples as an alternative to the traditional HAT field testing and confirmation approach. A cross-sectional study was conducted in HAT endemic and previously endemic zones in Kongo Central province, and a non-endemic zone in Haut Katanga province in the Democratic Republic of the Congo (DRC). Door-to-door visits were performed to collect dried blood spot (DBS) samples on filter paper. ELISA/T.b. gambiense was conducted followed by ITL for those testing positive by ELISA and in a subset of ELISA negatives. In total, 11,642 participants were enrolled. Of these, 11,535 DBS were collected and stored in appropriate condition for ELISA testing. Ninety-seven DBS samples tested positive on ELISA. In the endemic zone, ELISA positivity was 1.34% (95%CI: 1.04–1.64). In the previously endemic zone and non-endemic zone, ELISA positivity was 0.34% (95% CI: 0.13–0.55) and 0.37% (95% CI: 0.15–0.60) respectively. Among the ELISA positives, only two samples had a positive ITL result, both from the endemic zone. One of those was from a former HAT patient treated in 2008 and the other from an individual who unfortunately had deceased prior to the follow-up visit. Our study showed that a surveillance strategy, based on DBS samples and centralized testing with retracing of patients if needed, is feasible in DRC. ELISA seems well suited as initial test with a similar positivity rate as traditional screening tests, but ITL remains complex. Alternatives for the latter, also analyzable on DBS, should be further explored.

Abstract Although majority of chronic myeloid leukemia (CML) patients benefit from targeted therapy, there is an unmet need to identify as early as possible patients who develop resistance to tyrosine kinase inhibitors (TKI) and/or patients who progress to blastic phase (CML-BP). Searching for potential candidates of disease progression we have focused on BIRC (baculoviral IAP repeat-containing) genes expression in various stages of CML. This family includes eight functionally- and structurally-related proteins, most of them are believed to serve as endogenous inhibitors of apoptosis. Overexpression of various BIRC genes has been associated with cancer progression, multidrug resistance, poor prognosis and short survival in several types of neoplasms including hematological malignancies. In CML so far expression of BIRC5 (encoding survivin) and BIRC4 (encoding XIAP) has been associated with progression of the disease. However, there is no data on the role of other BIRC members in myeloproliferative diseases. To study the expression of BIRC genes in CML we employed RT-qPCR following MIQE guidelines. We analyzed sequential samples of peripheral blood leukocytes obtained from CML patients at various stages of the disease. Initially we looked at the samples from patients in chronic phase (CML-CP): at the diagnosis and after development of TKI resistance (confirmed as a loss of cytogenetic response, n=5). Then we analyzed samples from patients who progressed either to accelerated phase (CML-AccPh) or to blastic phase (CML-BP) (n=6). Among eight BIRC genes we observed significant decrease in BIRC3 (encoding cIAP2) and BIRC8 (encoding ILP2) expression. This was associated both with TKI resistance and with progression of CML to accelerated or blastic phase. Simultaneously, we observed marked increase in BIRC5 expression in samples from CML-AccPh and BP as compared to chronic phase (as was previously shown by others) but we observed no significant difference in BIRC5 expression between CML-CP diagnostic and TKI-resistant samples. Expression of BIRC1 (NAIP), BIRC2 (cIAP1), BIRC4 (XIAP), BIRC6 (BRUCE) and BIRC7 (LIVIN) was comparable in sequential samples from CML-CP and CML-BC and was not related to TKI-resistance. We verified these results by comparing larger group of patients in either CML-CP at the diagnosis prior to any treatment (n=15) or CML-BP (n=11). To compare the expression of BIRC family in CML to normal hematopoietic cells, we included also cDNA from healthy blood donors (n=10). In accordance with paired samples analysis, we observed downregulation of BIRC3 and BIRC8 expression in CML-BP (as compared to CML-CP and healthy blood donors), while BIRC5 was upregulated in CML-BP patients (as compared to CML-CP and healthy blood donors). There was no difference in expression of other BIRC family members. In conclusion, this is the first comprehensive analysis of the expression of all eight BIRC genes in the course of CML. In addition to the previously described upregulation of BIRC5, we observed significant downregulation of BIRC3 and BIRC8 associated with TKI-resistance and also with progression to accelerated or blastic phase. Recently Rossi D. et al. (Blood 2012;119: 2854-62) described potential role of disruption of BIRC3 (through mutation or gene deletion) in resistance to fludarabine in chronic lymphocytic leukemia. Together with our results this shows that current view linking BIRC genes upregulation with tumor progression and drug resistance may not be true for all of BIRC genes. Our results suggest novel and unexpected role of BIRC3 and BIRC8 in the clonal evolution of CML and open a new area for further exploration of the role of BIRC in CML progression. Disclosures: No relevant conflicts of interest to declare.

ABSTRACT The soil macrofauna plays an important role in the carbon and nitrogen cycle of terrestrial ecosystems. In order to gain more insight into the role of the intestinal microbiota in transformation and mineralization of organic matter during gut passage, we characterized the physicochemical conditions, microbial activities, and community structure in the gut of our model organism, the humus-feeding larva of the cetoniid beetle Pachnoda ephippiata. Microsensor measurements revealed an extreme alkalinity in the midgut, with highest values (pH > 10) between the second and third crown of midgut ceca. Both midgut and hindgut were largely anoxic, but despite the high pH, the redox potential of the midgut content was surprisingly high even in the largest instar. However, reducing conditions prevailed in the hindgut paunch of all instars (Eh ∼ −100 mV). Both gut compartments possessed a pronounced gut microbiota, with highest numbers in the hindgut, and microbial fermentation products were present in high concentrations. The stimulation of hindgut methanogenesis by exogenous electron donors, such as H2, formate, and methanol, together with considerable concentrations of formate in midgut and hemolymph, suggests that midgut fermentations are coupled to methanogenesis in the hindgut by an intercompartmental transfer of reducing equivalents via the hemolymph. The results of a cultivation-based enumeration of the major metabolic groups in midgut and hindgut, which yielded high titers of lactogenic, propionigenic, and acetogenic bacteria, are in good agreement not only with the accumulation of microbial fermentation products in the respective compartments but also with the results of a cultivation-independent characterization of the bacterial communities reported in the companion paper (M. Egert, B. Wagner, T. Lemke, A. Brune, and M. W. Friedrich, Appl. Environ. Microbiol. 69:6659-6668, 2003).

Hybrid simulation (HS) is an advanced simulation method that couples experimental testing and analytical modeling to better understand structural systems and individual components’ behavior under extreme events such as earthquakes. Conducting HS and real-time HS (RTHS) can be challenging with complex analytical substructures due to the nature of direct integration algorithms when the finite element method is employed. Thus, alternative methods such as machine learning (ML) models could help tackle these difficulties. This study aims to investigate the quality of the RTHS tests when a deep learning algorithm is used as a metamodel to represent the dynamic behavior of a nonlinear analytical substructure. The compact HS laboratory at the University of Nevada, Reno was utilized to conduct exclusive RTHS tests. Simulating a braced frame structure, the RTHS tests combined, for the first time, linear brace model specimens (physical substructure) along with nonlinear ML models for the frame (analytical substructure). Deep long short-term memory (Deep-LSTM) networks were employed and trained to develop the metamodels of the analytical substructure using the Python environment. The training dataset was obtained from pure analytical finite element simulations for the complete structure under earthquake excitation. The RTHS evaluations were first conducted for virtual RTHS tests, where substructuring was sought between the LSTM metamodel and virtual experimental substructure. To validate the proposed RTHS testing methodology and full system, several actual RTHS tests were conducted. The results from ML-based RTHS were evaluated for different ML models and compared against results from conventional RTHS with finite element models. The paper demonstrates the potential of conducting successful experimental RTHS using Deep-LSTM models, which could open the door for unparalleled new opportunities in structural systems design and assessment.

We characterized the extended substrate binding site of recombinant oligopeptidase B enzymes from Trypanosoma cruzi (Tc-OP) and Trypanosoma brucei (Tb-OP), evaluating the specificity of their S3, S2, S1′, S2′ and S3′ subsites. Five series of internally quenched fluorescent peptides based on the substrate Abz-AGGRGAQ-EDDnp [where Abz is o-aminobenzoic acid and EDDnp is N-(2,4-dinitrophenyl)ethylenediamine] were designed to contain amino acid residues with side chains of a minimum size, and each residue position of this substrate was modified. Synthetic peptides of different lengths derived from the human kininogen sequence were also examined, and peptides of up to 17 amino acids were found to be hydrolysed by Tc-OP and Tb-OP. These two oligopeptidases were essentially arginyl hydrolases, since for all peptides examined the only cleavage site was the Arg–Xaa bond. We also demonstrated that Tc-OP and Tb-OP have a very specific carboxypeptidase activity for basic amino acids, which depends on the presence of at least of a pair of basic amino acids at the C-terminal end of the substrate. The peptide with triple Arg residues (Abz-AGRRRAQ-EDDnp) was an efficient substrate for Tc-OP and Tb-OP: the Arg–Ala peptide bond was cleaved first and then two C-terminal Arg residues were successively removed. The S1′ subsite seems to be an important determinant of the specificity of both enzymes, showing a preference for Tyr, Ser, Thr and Gln as hydrogen donors. The presence of these amino acids at P1′ resulted in substrates that were hydrolysed with Km values in the sub-micromolar range. Taken together, this work supports the view that oligopeptidase B is a specialized protein-processing enzyme with a specific carboxypeptidase activity. Excellent substrates were obtained for Tb-OP and Tc-OP (Abz-AMRRTISQ-EDDnp and Abz-AHKRYSHQ-EDDnp respectively), which were hydrolysed with remarkably high kcat and low Km values.

Abstract Granulocyte-Colony stimulating factor (G-CSF) is widely used in the treatment of hematological malignancies. The administration of G-CSF is a novel therapy for ischemic heart disease (IHD), but current data are controversial and the safety and efficacy of G-CSF in acute or chronic IHD is unclear. Aim: to assess safety and efficacy of G-CSF administration and stem cell mobilization in patients with chronic refractory ‘no option’ IHD. Methods: After baseline cardiac assessment (CA) [Seattle Angina Questionnaire (SAQ), Bruce exercise stress test (EST), persantin-Sestamibi and dobutamine-echocardiographic imaging], stable ‘no option’ IHD patients received open-label G-CSF 10μg/kg for 5 days, with an EST (to facilitate myocardial cytokine generation and stem cell trafficking) on the 4th and 6th days. After 3 months, CA and the same regimen of G-CSF and ESTs was repeated, but in addition, leucopheresis and a randomized double-blinded intracoronary infusion of either CD133+ or unselected cells was performed (randomized data remains blinded). Final CA was 3 months thereafter. Results: Thirteen patients (12 male, 1 female, mean age 62) received 21 cycles of G-CSF. There were no deaths, Q-wave AMIs or any complications with long-term sequelae, although, transient troponin-I elevation (n = 3) and thrombocytopenia (n = 2) were observed. Mean CD133+ cell count rose from 1.28 to 56.12 x1012/L (p = 0.001). There was no age-related trend towards lower numbers of G-CSF mobilized CD34+ cells in the IHD patients, as compared to a group of younger (<60 y.o) normal male donors (n = 66). After the first cycle of G-CSF, SAQ angina frequency and physical limitation scores improved, and EST time increased (all p < 0.01). There was further but less marked improvement in each of these parameters after the second cycle of G-CSF and cell infusion. Overall, SAQ angina frequency score improved 46 points (95% CI +22 to +70, p = 0.003). This was reflected by reduced anginal frequency and nitrate use (both p < 0.005). Overall, SAQ physical limitation score improved 26 points (95% CI +17 to +35, p = 0.0003), EST time improved 97 seconds (95% CI +41 to +153, p = 0.005) and Duke Treadmill Score improved 4.3 points (95% CI −0.2 to +8.8, p = 0.058). Conclusions: G-CSF and intracoronary cell infusion is safe in chronic ‘no option’ IHD patients. In this phase I study G-CSF improved anginal frequency, nitrate use and EST performance. A placebo controlled phase II trial investigating G-CSF with an appropriate cytokine stimulus is warranted.

“A Clean, Well-lighted Place,” was first published in Scribner’s Magazine, in 1933, and ever since has been a significant focus of the literary world, with few daring to risk their literary aspirations, in rebuttal of previously published assertions as the dark forebodings of a world bereft of faith or joy. One is left to ponder a literary profession that appears bereft of a critical examination of the work, or, may not being able to see the forest for the trees. Responding to the assertion by Robert Penn Warren, that Hemingway lived in a “world of violent action,” WB Bache preferred to see the writer as a representative of unique craft and insight, and that he should be seen as, “a creative artist.” This is why I too, found Hemingway an enigma, as someone with a unique literary style, but possessing too, a wicked side. In his art, as he was in life, a hard drinking, womanizing, errant joker, who I feel certain here, is having a laugh at us all, from the other side. Sam Bluefarb (1971) wrote of the “Need to break through to some transcendent purpose—esthetic or religious—without which life seems to have little or no meaning.” Indeed, the melancholia which pervades this literary offering drags the reader down, into its darkness and despair, its depths of the maudlin, the mundane. The pathos may be evident, but does the meaning of the story have to be so dark, and so bitter? The “illogical dialogue sequence,” Warren Bennett (1990) ascribed to the tale, appears to be too bad, too lacking substance, too illogical for words, and so devoid of natural development that it takes on an artificiality such that it could only be a frolic that Hemingway is having, at our expense. Hemingway was a disciple of misogyny, this brute found love so often, not with docile, “pleasant”, or amenable women, but independent, vibrant, aggressive, articulate, intelligent, and yes, “feisty.” None of them was just a decoration, all were treated abysmally, and yet they all loved him till they had no more love left to give, until he had drained them of their capacity to continue to love him. These relationships open the door to a less discussed possibility, that “A Clean Well-lighted Place,” was actually a metaphoric celebration of femininity, in praise of womanhood, an explanation of the clean illumination of our lives (places), without whom, we are dark and dull, and lifeless, much like the iconic short story.

Abstract The apoptotic mode of cell death is a major regulatory process in all complex organisms. The slow accumulation of malignant cells in chronic lymphocytic leukemia (CLL) suggests that this disease is caused by a defect in apoptosis regulation. Apoptosis, defined as a programmed cell death, is executed through activity of caspases, cysteine proteases which are regulated by a number of pro- and anti-apoptotic proteins. One of such checkpoints is a control of caspase activation by the family of inhibitor of apoptosis proteins (IAPs). So far eight IAP proteins have been identified in human. They were XIAP, cIAP1, cIAP2, ILP-2, NAIP, survivin and BRUCE/Apollon. Moreover, an important role in the regulation of apoptosis play also three proteins which bind to IAPs and inhibit their activity, such as Smac/DIABLO, HtrA2/Omi and XAF1. Until recently IAPs and their antagonists were poorly researched in CLL. The majority of them were even not investigated in this disease yet. The primary aim of the study was to perform a complex analysis of expression of the IAPs family proteins and their antagonists in leukemic cells (ex vivo) in untreated CLL patients in comparison to the healthy controls. In this study we have also aimed to assess differences in expression of these proteins between patients with stable(SD) and progressive disease (PD). We tried to establish a potential prognostic significance of the IAPs expression and to investigate their correlation with the clinical course of the disease. The last aim was to analyze the expression of these proteins in the cell culture (in vitro settings) in response to the drugs routinely used in CLL. One hundred untreated patients with CLL were included in the study. Patients were divided into two groups: with SD (n=52) and PD (n=48). Twenty seven healthy donors served as a control group. Expression of IAPs and their inhibitors were investigated by flow cytometry, after careful comparative studies by Western blot and fluorescence microscopy. Protein expression was assessed on the basis of the mean fluorescence intensity (MFI). Results were compared with several disease-related parameters, such as clinical stage according to Rai, lymphocyte count, ZAP-70 and CD38 expression or serum beta-2-microglobulin concentration. The secondary endpoint of the study was to compare the results with the outcome of examined patients. Our study revealed significantly higher expression of cIAP1 (median MFI level: 81.1 vs 8.2; p=0.000001) and cIAP2 (median MFI level: 313.5 vs 146.8; p=0.014) and lower expression of IAP-antagonist - Smac/DIABLO (median MFI level: 147.9 vs 301.3; p=0.010) in the group of untreated CLL patients in comparison to the control group. Furthermore, patients with PD showed significantly higher expression of all analyzed IAPs compared with those with SD. Median MFI level for cIAP1 were: 131.2 vs 60.1; p=0.002, for cIAP2: 434.3 vs 301.5; p=0.026, for XIAP: 463.7 vs 225.4; p=0.002 and for survivin: 119.6 vs 38.6; p=0.00006. We also found a positive correlation between all examined protein’s expression and clinical stage according to Rai and high lymphocytosis, but only XIAP correlated with beta-2-microglobulin concentration (p=0.017). Moreover the study showed that simultaneous expression of two or more IAPs is associated with shorter overall survival. Additionally, analyzing the expression of IAPs and their antagonists in the cell cultures (in vitro) in response to cytotoxic drugs (Cladribine, Fludarabine, Mafosfamide) we observed significantly lower level of cIAP1, cIAP2 and XIAP as well as a higher level of proapoptotic Smac/DLABLO in comparison to the control, cytostatic-free cultures. In conclusion, CLL cells are characterized by overexpression of cIAP1 and cIAP2 and downregulation of Smac/DIABLO. The anti-apoptotic pattern of these apoptosis-regulating proteins expression may be responsible for pathological accumulation of malignant cells in CLL. Moreover, expression of all IAP proteins is significantly elevated in patients with PD, what indicates a role of inhibited proclivity of CLL cells to undergo spontaneous apoptosis in the disease progression. Importantly, the co-expression of two or more IAP proteins in tumor cell seems to be unfavorable prognostic factor in CLL patients. Thus, all those data suggest that IAPs/IAP-antagonists system plays an important role in the biology of CLL and further studies to confirm their prognostic usefulness are warranted.

Abstract Introduction Most genomic studies of classical Hodgkin lymphoma (CHL) have been confined to cell lines due to the difficulty of isolating sparsely distributed Hodgkin and Reed-Sternberg (HRS) cells from reactive background tissue. One approach to evaluate primary cases has been to microdissect HRS cells from fresh-frozen tissue biopsies, which has been used for gene expression profiling and array comparative genomic hybridization to assess copy number alterations (Hartmann et al., Haematologica 93, 2006; Brune et al, J Exp Med 205, 2008; Steidl et al., Blood 120, 2012; Steidl et al., Blood 116, 2010; Tiacci et al., Blood 120, 2012). However, laser capture microdissection is technically challenging, does not provide a pure tumor cell population, and yields very small amounts of nucleic acids that may not be adequate for deep sequencing. Typically, generating whole-exome sequence data from fewer than 104 cells, including single cells, requires whole genome amplification (WGA), and is of a quality suitable to detect large scale copy-number alterations, but not nucleotide level information to identify point mutations. Methods We used a flow cytometric cell isolation method, which has enabled rapid isolation of thousands of viable HRS cells from primary CHL tumors (Fromm, et al., Am J Clin Pathol 126, 2006). Here we developed a new ultra-low-input DNA exome sequencing protocol which we combined with flow cytometry using CD64, CD95, CD30, CD5, CD20, CD15, CD40, and CD45, to produce what is to our knowledge the first full exome deep sequencing study of primary cases of Hodgkin lymphoma. We obtained a sequence depth of over 10X with >90% coverage of the target exome in HRS, as well as tumor-infiltrating T cells (also sorted and used as somatic control), in ten primary cases of CHL and performed mutation, copy number variation, and loss-of-heterozygosity (LOH) analysis. Results We have identified 61 recurrent mutations, and 12 genes had somatic mutations in over 30% of cases. Non-synonymous or splice site mutations were seen in genes involved in antigen presentation (B2M), chromosome integrity (BCL7A), NF-kB activation (A20/TNFAIP3) and protein ubiquitination (HECW2 and UBE2A). We also obtained high-resolution copy number variation data indicating specific regions of gains and losses and intragenic chromosomal breakpoints. The most common genetic alterations from the combined analyses were in A20, present either as mutations or LOH, and alterations in b2-microglobulin (B2M), which were found in 80% of the cases sequenced. Alterations of B2M were inactivating and bi-allelic, leading to a lack of expression of MHC class I protein complex on the cell surface. The alterations include start codon mutations, exon-one splice donor site mutations, out of frame first-exon deletions and gene loss through chromosome-level deletion (LOH). Where possible, mutations were confirmed at the RNA level and resulted in lack of B2M protein expression documented by immunohistochemistry in an expanded cohort, where a total of 20 of 27 cases (74%) lacked B2M expression. Ectopic expression of B2M in a CHL cell line induced MHC class 1 expression, indicating that this genetic alteration is singly responsible for this defect in antigen presentation. In addition, B2M inactivation was highly prevalent in nodular sclerosis CHL, but was not found in any of the five cases of mixed cellularity CHL examined, indicating that these two types of CHL may belong to two different genetic categories. Conclusions We report the first exome deep sequencing of purified HRS cells from CHL tumor specimens, and reveal consistent alterations in important biological processes. The methodology developed allows exome sequencing from very low DNA input (10 ng), which has broader applications such as using FNA specimens from multiple tumor types. The genomic landscape of CHL revealed commonalities and differences with other lymphoma subtypes, like the consistent presence of A20 alterations that can lead to NF-kB activation. Inactivating mutations in B2M explain the complete lack of MHC class I expression, which likely affect the microenvironment. Since B2M expression is normal in all the cases evaluated that were histologically classified as mixed cellularity CHL, it is possible that this genetic alteration and resulting lack of B2M protein expression is a more accurate biomarker of CHL subtype than histological distinction. Disclosures: No relevant conflicts of interest to declare.

Abstract Abstract 3372 Poster Board III-260 Background: Although Hodgkin lymphoma is usually characterized by a high chemosensitivity and a good prognosis, a minority of patients with primary refractory disease or early relapse after intensive chemotherapy presents poor outcome (Brice. British Journal of Haematology 2008 ; 141 : 3-13. Moskowitz. British Journal of Haematology 2009 ; 146 : 158-163). AlloSCT is one of the treatment-options that can be proposed for these progressive patients but the optimal management remains controversial because comparative studies are lacking. However, several groups report encouraging survival results with acceptable non relapse mortality after alloSCT with reduced intensity conditioning. Method: We retrospectively studied 31 patients who received an alloSCT in Saint-Louis Hospital, Paris. The aim of the study was to compare their outcome with those of patients with similar characteristics who did not receive alloSCT. In order to have an homogenous group of patients, we only compared patients who relapsed after an autologous SCT (autoSCT) and were alive at least 12 months after autoSCT. To correct the recruitment bias, the propensity score (PS) methodology was used to determine the probability of receiving alloSCT in the group of patients who relapsed after autoSCT, according to a set of baseline characteristics (Rosenbaum. Biometrika 1983;70:41-55). Parameters used for this PS score were age, primary chemorefractory disease and stage III-IV at first relapse. This PS was estimated using logistic regression and used to match 1:1 patients with similar propensity to receive alloSCT, based on the nearest neighbor matching using calipers of width 0.2. AlloSCT recipients characteristics: Thirty-one patients (19 males and 12 females) received an alloSCT between 2000 and 2007. Median age at alloSCT was 26 years. Median number of previous lines of chemotherapy was 3.6 (range from 2 to 7) and 28 (90%) patients were previously autografted. 13 (42%) were considered refractory to primary treatment and 11 (35%) to secondary treatment. Status at alloSCT were: 20 (65%) patients in CR, 9 (29%) in PR and 2 (6%) in progressive disease. Pre-transplant co-morbidity score was 0 in 26 (84%), 1 in 3 (10%) and 2 in 2 (6%) patients. 26 (84%) patients received a reduced intensity conditioning regimen and TBI was performed in 23 (74%) recipients. Most of the patients (n=24, 77%) received peripheral stem cells and donors were HLA-identical siblings in 15 (48%) cases. Median follow-up after autoSCT and alloSCT was 62 and 51 months respectively. All patients engrafted. Grade II-IV acute GVHD occurred in 10 (32%) patients. Chronic GVHD occurred in 13 (42%). Three months after alloSCT, 19 patients were in remission, 5 presented stable disease and 7 were progressive. Eleven patients relapsed despite alloSCT and 11 patients died. The estimated 3-year overall and event free survival were respectively 69% (95% CI [48;82]) and 41% (95% CI [23;58]) and 3-year non relapse mortality was 21% (95% CI [13;29]). Causes of death after alloSCT were: progression in 6 (55%) and toxicity in 5 (45%) patients. From the 31 alloSCT recipients, 15 matched pairs were successfully constituted according to the PS. As expected, the distribution of age (p=0.97), stage III or IV at relapse (p=0.67) and primary chemorefractory disease (p=1) were not different between patients treated by alloSCT and untreated subjects. Based on these matched pairs, the estimated hazard ratio of death was 0.24 (95% CI [0.07;0.80]) for allografted patients compared with non allografted. In conclusion, alloSCT in poor-prognosis Hodgkin lymphoma gives promising survival outcomes although performed in advanced disease and provides survival benefit with acceptable non relapse mortality in comparison with non allograft treatment as suggested by PS method. Disclosures: No relevant conflicts of interest to declare.

Background: According to the United States Department of Health and Human services 8,497 liver transplants were accomplished in the United States in 2016. Of those transplants, only 345 (4.1%) of the transplants came from a live donor and only 9 (0.03%) of the live donor transplants came from a nonbiological paired donor [1]. Very little published data exists regarding Living Donor Liver Transplant (LDLT) influences on the live donor’s responses to exercise post-surgery. Purpose: To study the acute and prolonged effects of exercise and LDLT on heart rate, blood pressure, Lactate, Glucose, estimated maximal aerobic capacity, muscular strength, and the responses during submaximal exercise testing (6MWT and/or submaximal Bruce protocol). Methods: A single female subject, age 53 years, volunteered to participate in this study after already agreeing to become a live liver donor. Data was obtained once prior to the procedure and every month thereafter, for a total of eight months post-surgery. Conclusions: There was minimal changes in aerobic capacity and strength due to lack of consistency with an exercise program. Findings of this case report cannot be generalized to all LDLT donors. However, the information on the recovery of an LDLT donor in respect to exercise testing may be beneficial to clinicians and professionals in prescribing an exercise program for similar patients in similar circumstances.

The October 23, 2011 (Mw = 7.2) and November 9, 2011 (Mw = 5.6) earthquakes increased the damage in the minaret of Van Ulu Mosque, an important historical masonry structure built with solid bricks in Eastern Turkey, resulting in significant shear cracks. It was found that since the door and window openings are not symmetrically placed, they result in unsymmetrical stiffness distribution. The contribution of staircase and the core on stiffness is ignorable but its effect on the mass is significant. The pulpit with chamfered corner results in unsymmetrical transverse displacements. Brace wall improves the stiffness however contributes to the unsymmetrical behaviour considerably. The reason for the diagonal cracks can be attributed to the unsymmetrical brace wall and the chamfered pulpit but the effect of brace wall is more pronounced. After introducing the cracks, a new model was created and calibrated according to the results of Operational Modal Analysis. Diagonal cracks were found to be likely to develop under earthquake loading. Drifts are observed to increase significantly upon the introduction of the cracks.

Abstract Background Anterior cruciate ligament (ACL) injury can lead to reduced function, meniscal lesions, and early joint degeneration. Preservation of a torn ACL using the Internal Brace technique might re-establish normal knee kinematics, avoid donor-site morbidity due to tendon harvesting, and potentially maintain proprioception of the knee. Methods Fifty subjects were recruited for this study between December 2015 and October 2016. Two groups of individuals who sustained a unilateral ACL rupture were included: those who underwent surgery with preservation of the injured ACL (Internal Brace technique; IB) and those who underwent ACL reconstruction using a hamstring tendon graft (all-inside technique; AI). Subjective self-administered scores were used: the German version of the IKDC Subjective Knee Form (International Knee Documentation Committee), the German version of the WOMAC (Western Ontario and McMaster Universities Arthritis Index), SF-36 (short form), the German version of the KOOS (Knee Osteoarthritis Outcome Score), and the German version of themodified Lysholm Score by Lysholm and Gillquist. Anterior tibial translation was assessed using the KT-1000 Arthrometer (KT-1000 Knee Ligament Arthrometer, MEDmetric Corp., San Diego, CA, USA). Magnetic resonance evaluation was performed in all cases. Results Twenty-three subjects (46 %) were men, and the mean age was 34.7 years. The objective IKDC scores were “normal” in 15 and 14 patients, “nearly normal” in 11 and 7 patients, and “abnormal” in 1 and 2 patients, in the IB and AI groups, respectively. KT-1000 assessment showed a sideto-side difference of more than 3 mm on maximum manual testing in 11 (44 %) and 6 subjects (28.6 %) in the IB and AI groups, respectively. In the postoperative MRI, 20 (74 %) and 22 subjects (96 %) in the IB and AI groups had an intact ACL. Anterior tibial translation was significantly higher in the IB group compared with the AI group in the manual maximum test. Conclusions Preservation of the native ACL with the Internal Brace primary repair technique can achieve comparable results to ACL reconstruction using Hamstring autografts over a short term. Clinically relevant limitations such as a higher incidence of pathologic laxity, with patients more prone to pivot-shift phenomenon were observed during the study period.

Abstract Introduction The medial gastrocnemius flap is an established workhorse flap to cover proximal leg defects due to its reliability and simplicity to raise. However, it has the disadvantage of being bulky, requiring a skin graft for coverage, and is associated with loss of muscle power. The pedicled medial sural artery perforator (MSAP) flap has gained popularity as a reconstructive alternative for defects of the lower extremities. We present a case series of pedicled MSAP flaps for reconstructing defects around the knee as an alternative to the medial gastrocnemius flap. Materials and Methods A consecutive series of patients with proximal leg defects following trauma, osteomyelitis, burns, and chronic wounds were included. A hand-held Doppler was used to map out the MSAPs. Defects were reconstructed using pedicled MSAP flaps, preserving the nerve supply to the gastrocnemius muscle. Patient outcomes were recorded, including their Enneking scores postreconstruction. Results A total of 10 pedicled flaps was performed to reconstruct defects around the knee joint between October 2017 and November 2018. All the patients were discharged 1 week postoperatively, and rehabilitation consisted of graduated flexion in a knee brace by means of controlled passive mobilization. Three out of the ten patients developed complications: one patient developed flap congestion, one developed epidermolysis of the tip of the flap, and the other patient had partial necrosis of the skin paddle. The average Enneking score was 29 out of 35. Conclusion The pedicled MSAP flap is a good reconstructive option for proximal leg defects as it is associated with lower donor site morbidity and provides an aesthetically pleasing reconstruction.

This paper is part of a larger project exploring Australian literary responses to the Great War of 1914-1918. It draws on theories of embodiment, mourning, ritual and the recuperative potential of writing, together with a brief discussion of selected exemplars, to suggest that literary works of the period contain and lay bare a suite of creative, corporeal and social impulses, including resurrection, placation or stilling of ghosts, and formation of an empathic and duty-bound community. In Negotiating with the Dead, Margaret Atwood hypothesises that “all writing of the narrative kind, and perhaps all writing, is motivated, deep down, by a fear of and a fascination with mortality—by a desire to make the risky trip to the Underworld, and to bring something or someone back from the dead” (156). She asks an attendant question: “why should it be writing, over and above any other art or medium,” that functions this way? It is not only that writing acquires the appearance of permanence, by surviving “its own performance,” but also that some arts are transient, like dance, while others, like painting and sculpture and music, do “not survive as voice.” For Atwood, writing is a “score for voice,” and what the voice does mostly is tell stories, whether in prose or poetry: “Something unfurls, something reveals itself” (158). Writing, by this view, conjures, materialises or embodies the absent or dead, or is at least laden with this potential. Of course, as Katherine Sutherland observes, “representation is always the purview of the living, even when the order it constructs contains the dead” (202). She argues that all writing about death “might be regarded as epitaph or memorial; such writing is likely to contain the signs of ritual but also of ambiguity and forgetting” (204). Arguably writing can be regarded as participation in a ritual that “affirms membership of the collectivity, and through symbolic manipulation places the life of an individual within a much broader, sometimes cosmic, interpretive framework” (Seale 29), which may assist healing in relation to loss, even if some non-therapeutic purposes, such as restoration of social and political order, also lie behind both rites and writing. In a critical orthodoxy dating back to the 1920s, it has become accepted wisdom that the Australian literary response to the war was essentially nationalistic, “big-noting” ephemera, and thus of little worth (see Gerster and Caesar, for example). Consequently, as Bruce Clunies Ross points out, most Australian literary output of the period has “dropped into oblivion.” In his view, neglect of writings by First World War combatants is not due to its quality, “for this is not the only, or even the essential, condition” for consideration; rather, it is attributable to a “disjunction between the ideals enshrined in the Anzac legend and the experiences recorded or depicted” (170). The silence, we argue, also encompasses literary responses by non-combatants, many of whom were women, though limited space precludes consideration here of their particular contributions.Although poetry and fiction by those of middling or little literary reputation is not normally subject to critical scrutiny, it is patently not the case that there is no body of literature from the war period worthy of scholarly consideration, or that most works are merely patriotic, jingoistic, sentimental and in service of recruitment, even though these elements are certainly present. Our different proposition is that the “lost literatures” deserve attention for various reasons, including the ways they embody conflicting aims and emotions, as well as overt negotiations with the dead, during a period of unprecedented anguish. This is borne out by our substantial collection of creative writing provoked by the war, much of which was published by newspapers, magazines and journals. As Joy Damousi points out in The Labour of Loss, newspapers were the primary form of communication during the war, and never before or since have they dominated to such a degree; readers formed collective support groups through shared reading and actual or anticipated mourning, and some women commiserated with each other in person and in letters after reading casualty lists and death notices (21). The war produced the largest body count in the history of humanity to that time, including 60,000 Australians: none was returned to Australia for burial. They were placed in makeshift graves close to where they died, where possible marked by wooden crosses. At the end of the war, the Commonwealth War Graves Commission (CWGC) was charged with the responsibility of exhuming and reinterring bodily remains in immaculately curated cemeteries across Europe, at Gallipoli and in the Middle East, as if the peace demanded it. As many as one third of the customary headstones were inscribed with “known unto God,” the euphemism for bodies that could not be identified. The CWGC received numerous requests from families for the crosses, which might embody their loved one and link his sacrificial death with resurrection and immortality. For allegedly logistical reasons, however, all crosses were destroyed on site. Benedict Anderson suggested the importance to nationalism of the print media, which enables private reading of ephemera to generate a sense of communion with thousands or millions of anonymous people understood to be doing likewise. Furthermore, Judith Herman demonstrates in Trauma and Recovery that sharing traumatic experience with others is a “precondition for the restitution of a sense of a meaningful world” (70). Need of community and restitution extends to the dead. The practices of burying the dead together and of returning the dead to their homeland when they die abroad speak to this need, for “in establishing a society of the dead, the society of the living regularly recreates itself” (Hertz qtd. in Searle 66). For Australians, the society of the dead existed elsewhere, in unfamiliar terrain, accentuating the absence inherent in all death. The society of the dead and missing—and thus of the living and wounded—was created and recreated throughout the war via available means, including literature. Writers of war-related poems and fiction helped create and sustain imagined communities. Dominant use of conventional, sometimes archaic, literary forms, devices, language and imagery indicates desire for broadly accessible and purposeful communication; much writing invokes shared grief, resolve, gratitude, and sympathy. Yet, in many stories and poems, there is also ambivalence in relation to sacrifice and the community of the dead.Speaking in the voice of the other is a fundamental task of the creative writer, and the ultimate other, the dead, gaze upon and speak to or about the living in a number of poems. For example, they might vocalise displeasure and plead for reinforcements, as, for example, in Ella M’Fadyen’s poem “The Wardens,” published in the Sydney Mail in 1918, which includes the lines: “Can’t you hear them calling in the night-time’s lonely spaces […] Can’t you see them passing […] Those that strove full strongly, and have laid their lives away?” The speaker hears and conveys the pleading of those who have given their breath in order to make explicit the reader’s responsibility to both the dead and the Allied cause: “‘Thus and thus we battled, we were faithful in endeavour;/Still it lies unfinished—will ye make the deed in vain?’” M’Fadyen focusses on soldierly sacrifice and “drafts that never came,” whereas a poem entitled “Your Country’s Call,” published in the same paper in 1915 by “An Australian Mother, Shirley, Queensland,” refers to maternal sacrifice and the joys and difficulties of birthing and raising her son only to find the country’s claims on him outweigh her own. She grapples with patriotism and resistance: “he must go/forth./Where? Why? Don’t think. Just smother/up the pain./Give him up quickly, for his country’s gain.” The War Precautions Act of October 1914 made it “illegal to publish any material likely to discourage recruiting or undermine the Allied effort” (Damousi 21), which undoubtedly meant that, to achieve publication, critical, depressing or negative views would need to be repressed or cast as inducement to enlist, though evidently many writers also sought to convince themselves as well as others that the cause was noble and the cost redeemable. “Your Country’s Call” concludes uncertainly, “Give him up proudly./You have done your share./There may be recompense—somewhere.”Sociologist Clive Seal argues that “social and cultural life involves turning away from the inevitability of death, which is contained in the fact of our embodiment, and towards life” (1). He contends that “grief for embodiment” is pervasive and perpetual and “extends beyond the obvious manifestations of loss by the dying and bereaved, to incorporate the rituals of everyday interaction” (200), and he goes so far as to suggest that if we recognise that our bodies “give to us both our lives and our deaths” then we can understand that “social and cultural life can, in the last analysis, be understood as a human construction in the face of death” (210). To deal with the grief that comes with “realisation of embodiment,” Searle finds that we engage in various “resurrective practices designed to transform an orientation towards death into one that points towards life” (8). He includes narrative reconstruction as well as funeral lament and everyday conversation as rituals associated with maintenance of the social bond, which is “the most crucial human motive” (Scheff qtd. in Searle 30). Although Seale does not discuss the acts of writing or of reading specifically, his argument can be extended, we believe, to include both as important resurrective practices that contain desire for self-repair and reorientation as well as for inclusion in and creation of an empathic moral community, though this does not imply that such desires can ever be satisfied. In “Reading,” Virginia Woolf reminds that “somewhere, everywhere, now hidden, now apparent in whatever is written down is the form of a human being” (28-29), but her very reminder assumes that this knowledge of embodiment tends to be forgotten or repressed. Writing, by its aura of permanence and resurrective potential, points towards life and connection, even as it signifies absence and disconnection. Christian Riegel explains that the “literary work of mourning,” whether poetry, fiction or nonfiction, often has both a psychic and social function, “partaking of the processes of mourning while simultaneously being a product for public reception.” Such a text is indicative of ways that societies shape and control responses to death, making it “an inherently socio-historical construct” (xviii). Jacques Derrida’s passionate and uneasy enactment of this labour in The Work of Mourning suggests that writing often responds to the death of a known person or their oeuvre, where each death changes and reduces the world, so that the world as one knew it “sinks into an abyss” (115). Of course, writing also wrestles with anonymous, large-scale loss which is similarly capable of shattering our sense of “ontological security” (Riegel xx). Sandra Gilbert proposes that some traumatic events cause “death’s door” to swing “so publicly and dramatically open that we can’t look away” (xxii). Derrida’s work of mourning entails imaginative revival of those he has lost and is a struggle with representation and fidelity, whereas critical silence in respect of the body of literature of the First World War might imply repeated turning from “grief for embodiment” towards myths of immortality and indebtedness. Commemorating the war dead might be regarded as a resurrective practice that forges and fortifies communities of the living, while addressing the imagined demands of those who die for their nation.Riegel observes that in its multiplicity of motivations and functions, the literary work of mourning is always “an attempt to make present that which is irrefutably lost, and within that paradoxical tension lies a central tenet of all writerly endeavour that deals with the representation of death” (xix). The literary work of mourning must remain incomplete: it is “always a limiting attempt at revival and at representation,” because words inevitably “fail to replace a lost one.” Even so, they can assist in the attempt to “work through and understand” loss (xix). But the reader or mourner is caught in a strange situation, for he or she inevitably scrutinises words not the body, a corpus not a corpse, and while this is a form of evasion it is also the only possibility open to us. Even so, Derrida might say that it is “as if, by reading, by observing the signs on the drawn sheet of paper, [readers are] trying to forget, repress, deny, or conjure away death—and the anxiety before death.” But he also concedes (after Sarah Kofman), that this process might involve “a cunning affirmation of life, its irrepressible movement to survive, to live on” (176), which supports Seale’s contention in relation to resurrective practices generally. Atwood points out that the dead have always made demands on the living, but, because there is a risk in negotiating with the dead, there needs to be good reason or reward for doing so. Our reading of war literature written by noncombatants suggests that in many instances writers seek to appease the unsettled dead whose death was meant to mean something for the future: the living owe the dead a debt that can only be paid by changing the way they live. The living, in other words, must not only remember the fallen, but also heed them by their conduct. It becomes the poet’s task to remind people of this, that is, to turn them from death towards life.Arthur H Adams’s 1918 poem “When the Anzac Dead Came Home,” published in the Bulletin, is based on this premise: the souls of the dead— the “failed” and “fallen”—drift uncertainly over their homeland, observing the world to which they cannot return, with its “cheerful throng,” “fair women swathed in fripperies,” and “sweet girls” that cling “round windows like bees on honeycomb.” One soul recognises a soldier, Steve, from his former battalion, a mate who kept his life but lost his arm and, after hovering for a while, again “wafts far”; his homecoming creates a “strange” stabbing pain, an ache in his pal’s “old scar.” In this uncanny scene, irreconcilable and traumatic knowledge expresses itself somatically. The poet conveys the viewpoint of the dead Anzac rather than the returned one. The living soldier, whose body is a site of partial loss, does not explicitly conjure or mourn his dead friend but, rather, is a living extension of his loss. In fact, the empathic connection construed by the poet is not figured as spectral orchestration or as mindful on the part of man or community; rather, it occurs despite bodily death or everyday living and forgetting; it persists as hysterical pain or embodied knowledge. Freud and Breuer’s influential Studies on Hysteria, published in 1895, raised the issue of mind/body relations, given its theory that the hysteric’s body expresses psychic trauma that she or he may not recollect: repressed “memories of aetiological significance” result in “morbid symptoms” (56). They posited that experience leaves traces which, like disinterred archaeological artefacts, inform on the past (57). However, such a theory depends on what Rousseau and Porter refer to as an “almost mystical collaboration between mind and body” (vii), wherein painful or perverse or unspeakable “reminiscences” are converted into symptoms, or “mnemic symbols,” which is to envisage the body as penetrable text. But how can memory return unbidden and in such effective disguise that the conscious mind does not recognise it as memory? How can the body express pain without one remembering or acknowledging its origin? Do these kinds of questions suggest that the Cartesian mind/body split has continued valency despite the challenge that hysteria itself presents to such a theory? Is it possible, rather, that the body itself remembers—and not just its own replete form, as suggested by those who feel the presence of a limb after its removal—but the suffering body of “the other”? In Adam’s poem, as in M’Fadyen’s, intersubjective knowledge subsists between embodied and disembodied subjects, creating an imagined community of sensation.Adams’s poem envisions mourning as embodied knowledge that allows one man to experience another’s pain—or soul—as both “old” and “strange” in the midst of living. He suggests that the dead gaze at us even as they are present “in us” (Derrida). Derrida reminds that ghosts occupy an ambiguous space, “neither life nor death, but the haunting of the one by the other” (41). Human mutability, the possibility of exchanging places in a kind of Socratic cycle of life and death, is posited by Adams, whose next stanzas depict the souls of the war dead reclaiming Australia and displacing the thankless living: blown to land, they murmur to each other, “’Tis we who are the living: this continent is dead.” A significant imputation is that the dead must be reckoned with, deserve better, and will not rest unless the living pay their moral dues. The disillusioned tone and intent of this 1918 poem contrasts with a poem Adams published in the Bulletin in 1915 entitled “The Trojan War,” which suggests even “Great Agamemnon” would “lift his hand” to honour “plain Private Bill,” the heroic, fallen Anzac who ventured forth to save “Some Mother-Helen sad at home. Some obscure Helen on a farm.” The act of war is envisaged as an act of birthing the nation, anticipating the Anzac legend, but simultaneously as its epitaph: “Upon the ancient Dardanelles New peoples write—in blood—their name.” Such a poem arguably invokes, though in ambiguous form, what Derrida (after Lyotard) refers to as the “beautiful death,” which is an attempt to lift death up, make it meaningful, and thereby foreclose or limit mourning, so that what threatens disorder and despair might instead reassure and restore “the body politic,” providing “explicit models of virtue” (Nass 82-83) that guarantee its defence and survival. Adams’ later poem, in constructing Steve as “a living fellow-ghost” of the dead Anzac, casts stern judgement on the society that fails to notice what has been lost even as it profits by it. Ideological and propagandist language is also denounced: “Big word-warriors still played the Party game;/They nobly planned campaigns of words, and deemed/their speeches deeds,/And fought fierce offensives for strange old creeds.” This complaint recalls Ezra Pound’s lines in Hugh Selwyn Mauberley about the dead who “walked eye-deep in hell/believing in old men’s lies, then unbelieving/came home, home to a lie/home to many deceits,/home to old lies and new infamy;/usury age-old and age-thick/and liars in public places,” and it would seem that this is the kind of disillusion and bitterness that Clunies Ross considers to be “incompatible with the Anzac tradition” (178) and thus ignored. The Anzac tradition, though quieted for a time, possibly due to the 1930s Depression, Second World War, Vietnam War and other disabling events has, since the 1980s, been greatly revived, with Anzac Day commemorations in Australia and at Gallipoli growing exponentially, possibly making maintenance of this sacrificial national mythology, or beautiful death, among Australia’s most capacious and costly creative industries. As we approach the centenary of the war and of Gallipoli, this industry will only increase.Elaine Scarry proposes that the imagination invents mechanisms for “transforming the condition of absence into presence” (163). It does not escape us that in turning towards lost literatures we are ourselves engaging in a form of resurrective practice and that this paper, like other forms of social and cultural practice, might be understood as one more human construction motivated by grief for embodiment.Note: An archive and annotated bibliography of the “Lost Literatures of the First World War,” which comprises over 2,000 items, is expected to be published online in 2015.References Adams, Arthur H. “When the Anzac Dead Came Home.” Bulletin 21 Mar. 1918.---. “The Trojan War.” Bulletin 20 May 1915.An Australian Mother. “Your Country’s Call.” Sydney Mail 19 May 1915.Anderson, Benedict. Imagined Communities: Reflections on the Origin and Spread of Nationalism. 2nd ed. London: Verso, 1991.Atwood, Margaret. Negotiating with the Dead: A Writer on Writing. New York: Random House, 2002.Caesar, Adrian. “National Myths of Manhood: Anzac and Others.” The Oxford Literary History of Australia. Eds. Bruce Bennett and Jennifer Strauss. Melbourne: Oxford University Press, 1998. 147-168.Clunies Ross, Bruce. “Silent Heroes.” War: Australia’s Creative Response. Eds. Anna Rutherford and James Wieland. West Yorkshire: Dangaroo Press, 1997. 169-181.Damousi, Joy. The Labour of Loss: Mourning, Memory and Wartime Bereavement in Australia. Cambridge: Cambridge UP, 1999.Derrida, Jacques. The Work of Mourning. Chicago: University of Chicago Press, 2001.Freud, Sigmund, and Joseph Breuer. Studies on Hysteria. Pelican Freud Library. Vol. 3. Trans. and eds. James Strachey, Alix Strachey, and Angela Richards. London: Penguin, 1988.Gerster, Robin. Big Noting: The Heroic Theme in Australian War Writing. Melbourne: Melbourne University Press, 1992.Gilbert, Sandra M. Death’s Door: Modern Dying and the Ways We Grieve. New York: W.W. Norton, 2006.Herman, Judith. Trauma and Recovery. New York: Basic Books, 1992. M’Fayden, Ella. “The Wardens.” Sydney Mail 17 Apr. 1918.Naas, Michael. “History’s Remains: Of Memory, Mourning, and the Event.” Research in Phenomenology 33 (2003): 76-96.Pound, Ezra. “Hugh Selwyn Mauberly.” iv. 1920. 19 June 2012. ‹http://www.archive.org/stream/hughselwynmauber00pounrich/hughselwynmauber00pounrich_djvu.txt›.Riegal, Christian, ed. Response to Death: The Literary Work of Mourning. Edmonton, Alberta: University of Alberta Press, 2005. Rousseau, G.S., and Roy Porter. “Introduction: The Destinies of Hysteria.” Hysteria beyond Freud. Ed. Sander L. Gilman, Helen King, Roy Porter, G.S. Rousseau, and Elaine Showalter. Berkeley: University of California Press, 1993.Scarry, Elaine. The Body in Pain: The Making and Unmaking of the World. Oxford: Oxford University Press, 1985.Seale, Clive. Constructing Death: The Sociology of Dying and Bereavement. Cambridge: Cambridge University Press, 1998.Sutherland, Katherine. “Land of Their Graves: Maternity, Mourning and Nation in Janet Frame, Sara Suleri, and Arundhati Roy.” Riegel 201-16.Woolf, Virginia. Collected Essays Volume 2. London: Hogarth, 1966. 28-29.

1909–22: Turkey exterminated over 1.5 million of its ethnically Armenian, and hundreds of thousands of its ethnically Greek and Assyrian, citizens. Most died in 1915. This period of decimation in now widely called the Armenian Genocide (Balakian 179-80).1910: Siamanto first published his poem, The Dance: “The corpses were piled as trees, / and from the springs, from the streams and the road, / the blood was a stubborn murmur.” When springs run red, when the dead are stacked tree-high, when “everything that could happen has already happened,” then time is nothing: “there is no future [and] the language of civilised humanity is not our language” (Nichanian 142).2007: In my novel The Edge of the World a ceramic bowl, luminous blue, recurs as motif. Imagine you are tiny: the bowl is broken but you don’t remember breaking it. You’re awash with tears. You sit on the floor, gather shards but, no matter how you try, you can’t fix it. Imagine, now, that the bowl is the sky, huge and upturned above your head. You have always known, through every wash of your blood, that life is shockingly precarious. Silence—between heartbeats, between the words your parents speak—tells you: something inside you is terribly wrong; home is not home but there is no other home; you “can never be fully grounded in a community which does not share or empathise with the experience of persecution” (Wajnryb 130). This is the stubborn murmur of your body.Because time is nothing, this essay is fragmented, non-linear. Its main characters: my mother, grandmother (Hovsanna), grandfather (Benyamin), some of my mother’s older siblings (Krikor, Maree, Hovsep, Arusiak), and Mustafa Kemal Ataturk (Ottoman military officer, Young Turk leader, first president of Turkey). 1915–2013: Turkey invests much energy in genocide denial, minimisation and deflection of responsibility. 24 April 2012: Barack Obama refers to the Medz Yeghern (Great Calamity). The use of this term is decried as appeasement, privileging political alliance with Turkey over human rights. 2003: Between Genocide and Catastrophe, letters between Armenian-American theorist David Kazanjian and Armenian-French theorist Marc Nichanian, contest the naming of the “event” (126). Nichanian says those who call it the Genocide are:repeating every day, everywhere, in all places, the original denial of the Catastrophe. But this is part of the catastrophic structure of the survivor. By using the word “Genocide”, we survivors are only repeating […] the denial of the loss. We probably cannot help it. We are doing what the executioner wanted us to do […] we claim all over the world that we have been “genocided;” we relentlessly need to prove our own death. We are still in the claws of the executioner. We still belong to the logic of the executioner. (127)1992: In Revolution and Genocide, historian Robert Melson identifies the Armenian Genocide as “total” because it was public policy intended to exterminate a large fraction of Armenian society, “including the families of its members, and the destruction of its social and cultural identity in most or all aspects” (26).1986: Boyajian and Grigorian assert that the Genocide “is still operative” because, without full acknowledgement, “the ghosts won’t go away” (qtd. in Hovannisian 183). They rise up from earth, silence, water, dreams: Armenian literature, Armenian homes haunted by them. 2013: My heart pounds: Medz Yeghern, Aksor (Exile), Anashmaneli (Indefinable), Darakrutiun (Deportation), Chart (Massacre), Brnagaght (Forced migration), Aghed (Catastrophe), Genocide. I am awash. Time is nothing.1909–15: Mustafa Kemal Ataturk was both a serving Ottoman officer and a leader of the revolutionary Young Turks. He led Ottoman troops in the repulsion of the Allied invasion before dawn on 25 April at Gallipoli and other sites. Many troops died in a series of battles that eventually saw the Ottomans triumph. Out of this was born one of Australia’s founding myths: Australian and New Zealand Army Corps (ANZACs), courageous in the face of certain defeat. They are commemorated yearly on 25 April, ANZAC Day. To question this myth is to risk being labelled traitor.1919–23: Ataturk began a nationalist revolution against the occupying Allies, the nascent neighbouring Republic of Armenia, and others. The Allies withdrew two years later. Ataturk was installed as unofficial leader, becoming President in 1923. 1920–1922: The last waves of the Genocide. 2007: Robert Manne published A Turkish Tale: Gallipoli and the Armenian Genocide, calling for a recontextualisation of the cultural view of the Gallipoli landings in light of the concurrence of the Armenian Genocide, which had taken place just over the rise, had been witnessed by many military personnel and widely reported by international media at the time. Armenian networks across Australia were abuzz. There were media discussions. I listened, stared out of my office window at the horizon, imagined Armenian communities in Sydney and Melbourne. Did they feel like me—like they were holding their breath?Then it all went quiet. Manne wrote: “It is a wonderful thing when, at the end of warfare, hatred dies. But I struggle to understand why Gallipoli and the Armenian Genocide continue to exist for Australians in parallel moral universes.” 1992: I bought an old house to make a home for me and my two small children. The rooms were large, the ceilings high, and behind it was a jacaranda with a sturdy tree house built high up in its fork. One of my mother’s Armenian friends kindly offered to help with repairs. He and my mother would spend Saturdays with us, working, looking after the kids. Mum would stay the night; her friend would go home. But one night he took a sleeping bag up the ladder to the tree house, saying it reminded him of growing up in Lebanon. The following morning he was subdued; I suspect there were not as many mosquitoes in Lebanon as we had in our garden. But at dinner the previous night he had been in high spirits. The conversation had turned, as always, to politics. He and my mother had argued about Turkey and Russia, Britain’s role in the development of the Middle East conflict, the USA’s roughshod foreign policy and its effect on the world—and, of course, the Armenian Genocide, and the killingof Turkish governmental representatives by Armenians, in Australia and across the world, during the 1980s. He had intimated he knew the attackers and had materially supported them. But surely it was the beer talking. Later, when I asked my mother, she looked at me with round eyes and shrugged, uncharacteristically silent. 2002: Greek-American diva Diamanda Galas performed Dexifiones: Will and Testament at the Perth Concert Hall, her operatic work for “the forgotten victims of the Armenian and Anatolian Greek Genocide” (Galas).Her voice is so powerful it alters me.1925: My grandmother, Hovsanna, and my grandfather, Benyamin, had twice been separated in the Genocide (1915 and 1922) and twice reunited. But in early 1925, she had buried him, once a prosperous businessman, in a swamp. Armenians were not permitted burial in cemeteries. Once they had lived together in a big house with their dozen children; now there were only three with her. Maree, half-mad and 18 years old, and quiet Hovsep, aged seven,walked. Then five-year-old aunt, Arusiak—small, hungry, tired—had been carried by Hovsanna for months. They were walking from Cilicia to Jerusalem and its Armenian Quarter. Someone had said they had seen Krikor, her eldest son, there. Hovsanna was pregnant for the last time. Together the four reached Aleppo in Syria, found a Christian orphanage for girls, and Hovsanna, her pregnancy near its end, could carry Arusiak no further. She left her, promising to return. Hovsanna’s pains began in Beirut’s busy streets. She found privacy in the only place she could, under a house, crawled in. Whenever my mother spoke of her birth she described it like this: I was born under a stranger’s house like a dog.1975: My friend and I travelled to Albany by bus. After six hours we were looking down York Street, between Mount Clarence and Mount Melville, and beyond to Princess Royal Harbour, sapphire blue, and against which the town’s prosperous life—its shopfronts, hotels, cars, tourists, historic buildings—played out. It took away my breath: the deep harbour, whaling history, fishing boats. Rain and sun and scudding cloud; cliffs and swells; rocky points and the white curves of bays. It was from Albany that young Western Australian men, volunteers for World War I, embarked on ships for the Middle East, Gallipoli, sailing out of Princess Royal Harbour.1985: The Australian Government announced that Turkey had agreed to have the site of the 1915 Gallipoli landings renamed Anzac Cove. Commentators and politicians acknowledged it as historic praised Turkey for her generosity, expressed satisfaction that, 70 years on, former foes were able to embrace the shared human experience of war. We were justifiably proud of ourselves.2005: Turkey made her own requests. The entrance to Albany’s Princess Royal Harbour was renamed Ataturk Channel. A large bronze statue of Ataturk was erected on the headland overlooking the Harbour entrance. 24 April 1915: In the town of Hasan Beyli, in Cilicia, southwest Turkey, my great grandfather, a successful and respected businessman in his 50s, was asleep in his bed beside his wife. He had been born in that house, as had his father, grandfather, and all his children. His brother, my great uncle, had bought the house next door as a young man, brought his bride home to it, lived there ever since; between the two households there had been one child after another. All the cousins grew up together. My great grandfather and great uncle had gone to work that morning, despite their wives’ concerns, but had returned home early. The women had been relieved to see them. They made coffee, talked. Everyone had heard the rumours. Enemy ships were massing off the coast. 1978: The second time in Albany was my honeymoon. We had driven into the Goldfields then headed south. Such distance, such beautiful strangeness: red earth, red rocks; scant forests of low trees, thin arms outstretched; the dry, pale, flat land of Norseman. Shimmering heat. Then the big, wild coast.On our second morning—a cool, overcast day—we took our handline to a jetty. The ocean was mercury; a line of cormorants settled and bobbed. Suddenly fish bit; we reeled them in. I leaned over the jetty’s side, looked down into the deep. The water was clear and undisturbed save the twirling of a pike that looked like it had reversed gravity and was shooting straight up to me. Its scales flashed silver as itbroke the surface.1982: How could I concentrate on splicing a film with this story in my head? Besides the desk, the only other furniture in the editing suite was a whiteboard. I took a marker and divided the board into three columns for the three generations: my grandparents, Hovsanna and Benyamin; my mother; someone like me. There was a lot in the first column, some in the second, nothing in the third. I stared at the blankness of my then-young life.A teacher came in to check my editing. I tried to explain what I had been doing. “I think,” he said, stony-faced, “that should be your third film, not your first.”When he had gone I stared at the reels of film, the white board blankness, the wall. It took 25 years to find the form, the words to say it: a novel not a film, prose not pictures.2007: Ten minutes before the launch of The Edge of the World, the venue was empty. I made myself busy, told myself: what do you expect? Your research has shown, over and over, this is a story about which few know or very much care, an inconvenient, unfashionable story; it is perfectly in keeping that no-one will come. When I stepped onto the rostrum to speak, there were so many people that they crowded the doorway, spilled onto the pavement. “I want to thank my mother,” I said, “who, pretending to do her homework, listened instead to the story her mother told other Armenian survivor-women, kept that story for 50 years, and then passed it on to me.” 2013: There is a section of The Edge of the World I needed to find because it had really happened and, when it happened, I knew, there in my living room, that Boyajian and Grigorian (183) were right about the Armenian Genocide being “still operative.” But I knew even more than that: I knew that the Diaspora triggered by genocide is both rescue and weapon, the new life in this host nation both sanctuary and betrayal. I picked up a copy, paced, flicked, followed my nose, found it:On 25 April, the day after Genocide memorial-day, I am watching television. The Prime Minister stands at the ANZAC memorial in western Turkey and delivers a poetic and moving speech. My eyes fill with tears, and I moan a little and cover them. In his speech he talks about the heroism of the Turkish soldiers in their defence of their homeland, about the extent of their losses – sixty thousand men. I glance at my son. He raises his eyebrows at me. I lose count of how many times Kemal Ataturk is mentioned as the Father of Modern Turkey. I think of my grandmother and grandfather, and all my baby aunts and uncles […] I curl over like a mollusc; the ache in my chest draws me in. I feel small and very tired; I feel like I need to wash.Is it true that if we repeat something often enough and loud enough it becomes the truth? The Prime Minister quotes Kemal Ataturk: the ANZACS who died and are buried on that western coast are deemed ‘sons of Turkey’. My son turns my grandfather’s, my mother’s, my eyes to me and says, It is amazing they can be so friendly after we attacked them.I draw up my knees to my chest, lay my head and arms down. My limbs feel weak and useless. My throat hurts. I look at my Australian son with his Armenian face (325-6).24 April 1915 cont: There had been trouble all my great grandfather’s life: pogrom here, massacre there. But this land was accustomed to colonisers: the Mongols, the Persians, latterly the Ottomans. They invade, conquer, rise, fall; Armenians stay. This had been Armenian homeland for thousands of years.No-one masses ships off a coast unless planning an invasion. So be it. These Europeans could not be worse than the Ottomans. That night, were my great grandfather and great uncle awoken by the pounding at each door, or by the horses and gendarmes’ boots? They were seized, each family herded at gunpoint into its garden, and made to watch. Hanging is slow. There could be no mistakes. The gendarmes used the stoutest branches, stayed until they were sure the men weredead. This happened to hundreds of prominent Armenian men all over Turkey that night.Before dawn, the Allies made landfall.Each year those lost in the Genocide are remembered on 24 April, the day before ANZAC Day.1969: I asked my mother if she had any brothers and sisters. She froze, her hands in the sink. I stared at her, then slipped from the room.1915: The Ottoman government decreed: all Armenians were to surrender their documents and report to authorities. Able-bodied men were taken away, my grandfather among them. Women and children, the elderly and disabled, were told to prepare to walk to a safe camp where they would stay for the duration of the war. They would be accompanied by armed soldiers for their protection. They were permitted to take with them what they could carry (Bryce 1916).It began immediately, pretty young women and children first. There are so many ways to kill. Months later, a few dazed, starved survivors stumbled into the Syrian desert, were driven into lakes, or herded into churches and set alight.Most husbands and fathers were never seen again. 2003: I arrived early at my son’s school, parked in the shade, opened The Silence: How Tragedy Shapes Talk, and began to read. Soon I was annotating furiously. Ruth Wajnryb writes of “growing up among innocent peers in an innocent landscape” and also that the notion of “freedom of speech” in Australia “seems often, to derive from that innocent landscape where reside people who have no personal scars or who have little relevant historical knowledge” (141).1984: I travelled to Vancouver, Canada, and knocked on Arusiak’s door. Afraid she would not agree to meet me, I hadn’t told her I was coming. She was welcoming and gracious. This was my first experience of extended family and I felt loved in a new and important way, a way I had read about, had observed in my friends, had longed for. One afternoon she said, “You know our mother left me in an orphanage…When I saw her again, it was too late. I didn’t know who they were, what a family was. I felt nothing.” “Yes, I know,” I replied, my heart full and hurting. The next morning, over breakfast, she quietly asked me to leave. 1926: When my mother was a baby, her 18 year-old sister, Maree, tried to drown her in the sea. My mother clearly recalled Maree’s face had been disfigured by a sword. Hovsanna, would ask my mother to forgive Maree’s constant abuse and bad behaviour, saying, “She is only half a person.”1930: Someone gave Hovsanna the money to travel to Aleppo and reclaim Arusiak, by then 10 years old. My mother was intrigued by the appearance of this sister but Arusiak was watchful and withdrawn. When she finally did speak to my then five-year-old mother, she hissed: “Why did she leave me behind and keep you?”Soon after Arusiak appeared, Maree, “only half a person,” disappeared. My mother was happy about that.1935: At 15, Arusiak found a live-in job and left. My mother was 10 years old; her brother Hovsep, who cared for her before and after school every day while their mother worked, and always had, was seventeen. She adored him. He had just finished high school and was going to study medicine. One day he fell ill. He died within a week.1980: My mother told me she never saw her mother laugh or, once Hovsep died, in anything other than black. Two or three times before Hovsep died, she saw her smile a little, and twice she heard her singing when she thought she was alone: “A very sad song,” my mother would say, “that made me cry.”1942: At seventeen, my mother had been working as a live-in nanny for three years. Every week on her only half-day off she had caught the bus home. But now Hovsanna was in hospital, so my mother had been visiting her there. One day her employer told her she must go to the hospital immediately. She ran. Hovsanna was lying alone and very still. Something wasn’t right. My mother searched the hospital corridors but found no-one. She picked up a phone. When someone answered she told them to send help. Then she ran all the way home, grabbed Arusiak’s photograph and ran all the way back. She laid it on her mother’s chest, said, “It’s all right, Mama, Arusiak’s here.”1976: My mother said she didn’t like my boyfriend; I was not to go out with him. She said she never disobeyed her own mother because she really loved her mother. I went out with my boyfriend. When I came home, my belongings were on the front porch. The door was bolted. I was seventeen.2003: I read Wajnryb who identifies violent eruptions of anger and frozen silences as some of the behaviours consistent in families with a genocidal history (126). 1970: My father had been dead over a year. My brothers and I were, all under 12, made too much noise. My mother picked up the phone: she can’t stand us, she screamed; she will call an orphanage to take us away. We begged.I fled to my room. I couldn’t sit down. I couldn’t keep still. I paced, pressed my face into a corner; shook and cried, knowing (because she had always told us so) that she didn’t make idle threats, knowing that this was what I had sometimes glimpsed on her face when she looked at us.2012: The Internet reveals images of Ataturk’s bronze statue overlooking Princess Royal Harbour. Of course, it’s outsized, imposing. The inscription on its plinth reads: "Peace at Home/ Peace in the World." He wears a suit, looks like a scholar, is moving towards us, a scroll in his hand. The look in his eyes is all intensity. Something distant has arrested him – a receding or re-emerging vision. Perhaps a murmur that builds, subsides, builds again. (Medz Yeghern, Aksor, Aghed, Genocide). And what is written on that scroll?2013: My partner suggested we go to Albany, escape Perth’s brutal summer. I tried to explain why it’s impossible. There is no memorial in Albany, or anywhere else in Western Australia, to the 1.5 million victims of the Armenian Genocide. ReferencesAkcam, Taner. “The Politics of Genocide.” Online Video Clip. YouTube. YouTube, 11 Dec. 2011. 6 Mar. 2013 ‹http://www.youtube.com/watchv=OxAJaaw81eU&noredirect=1genocide›.Balakian, Peter. The Burning Tigress: The Armenian Genocide. 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