Which Version™? Exploring the digital modelling of architecturally important sites and the implications on the understanding of the historical narrative the model reflects

The preservation of architecturally important buildings and monuments takes on various forms, with the reasons for the need to retain them varying from region to region, nation by nation. The parameters and criteria for this preservation can be informed by politics, economic considerations, the prevailing zeitgeist etc. Similarly, the wanton destruction of architecture can be influenced by numerous agendas, with recent examples including the destruction of the monumental Buddha statues in Afghanistan by the Taliban and the damage to the ancient city of Palmyra by daesh during the ongoing Syrian civil war. With the destruction of architecturally important buildings and monuments, the photographic, archaeological and academic studies related to them have been used to restore as much of the original form as possible. With the advent of digital methods of recording and creating Computer Aided Design models, with the capacity for producing virtual immersive versions, the capacity to both preserve and then share remotely those buildings expands. With particular reference to Palmyra, a city very much on the front line with no guarantee Daesh will not recapture it and recommence their deliberate, targeted cultural destruction, digital models may be the only way to study the city, and preserve it, for the foreseeable future. Cultural vandalism, the destruction or modification of architecture and monuments, the wanton eradication of buildings considered important to some but inconvenient blocks to further development to others, are part of architectural history’s long story. Other instances, such as the destruction of buildings associated with the Nazi regime, can be seen as part of a cathartic exercise of renewal and understandable revenge. However, versions of these buildings exist in fiction and museum models and are likely the source of veneration for those of a particular political perspective. With the range of complexities surrounding what is preserved and why it is preserved, there are questions which arise as to who has control over the preserved model, what time period does it cover, how is ‘Disneyfication’, or the ‘Vegas’ version avoided etc. These will be explored as part of this paper with its aim being the examination of the themes set out here to begin a conversation on how sites like Palmyra can be preserved and recorded. For example, the restoration of Palmyra to a pre-daesh state could arguably include the Tadmur prison, a notorious site for alleged torture since the Assad family came to power. Is it an honest reflection of modern Palmyra, a site made up of influences from civilisations across a 2,000-year period, if its modern reflection as an example of the failure of civilisation is not also recognised and preserved? As will be set out in the emerging conclusions of the paper, the importance of establishing a multifaceted context, and engaging in an inclusive dialogue with all the affected parties is key to attempting to establish What Version™ eventually emerges

Methyl jasmonate-elicited herbivore resistance: does MeJA function as a signal without being hydrolyzed to JA?

Treatment with methyl jasmonate (MeJA) elicits herbivore resistance in many plant species and over-expression of JA carboxyl methyltransferase (JMT) constitutively increases JA-induced responses in Arabidopsis. When wild-type (WT) Nicotiana attenuata plants are treated with MeJA, a rapid transient endogenous JA burst is elicited, which in turn increases levels of nicotine and trypsin proteinase inhibitors (TPIs) and resistance to larvae of the specialist herbivore, Manduca sexta. All of these responses are impaired in plants silenced in lipoxygenase 3 expression (asLOX3) but are restored to WT levels by MeJA treatment. Whether these MeJA-induced responses are directly elicited by MeJA or by its cleavage product, JA, is unknown. Using virus-induced gene silencing (VIGS), we silenced MeJA-esterase (NaMJE) expression and found this gene responsible for most of the MeJA-cleaving activity in N. attenuata protein extracts. Silencing NaMJE in asLOX3, but not in WT plants, significantly reduced MeJA-induced nicotine levels and resistance to M. sexta, but not TPI levels. MeJA-induced transcript levels of threonine deaminase (NaTD) and phenylalanine ammonia lyase (NaPAL1) were also decreased in VIGS MJE (asLOX3) plants. Finally the performance of M. sexta larvae that fed on plants treated with JA or MeJA demonstrated that silencing NaMJE inhibited MeJA-induced but not JA-induced resistance in asLOX3 plants. From these results, we conclude that the resistance elicited by MeJA treatment is directly elicited not by MeJA but by its de-methylated product, JA

Purification, cloning and heterologous expression of methyl jasmonate esterase from Lycopersicon esculentum

Aus Lycopersicon esculentum Zellsuspensionskulturen konnte ein bisher unbekanntes Enzym isoliert und beschrieben werden, das die Hydrolyse von Methyljasmonat (MeJA) zu Jasmonsäure (JA) katalysiert. Das Enzym wurde als Methyljasmonat-Esterase (MeJA-Esterase) bezeichnet. Mittels Methyl-[2-14C]JA und [Methyl-3H]MeJA wurden qualitative und quantitative Enzymtestsysteme etabliert, welche die Reinigung und Charakterisierung des Enzyms erlaubten. Methyljasmonat-Esterase Aktivität konnte in 18 taxonomisch unterschiedlichen Zellsuspensionskulturen höherer Pflanzen sowie in differenziertem Gewebe (Blüte, Wurzel, Stengel und Blatt) von Lycopersicon esculentum cv. Moneymaker nachgewiesen werden. In einem 6-stufigen Reinigungsverfahren wurde das native Enzym mit einer Ausbeute von 2.2 % bis zur Homogenität 767-fach angereichert. Die native MeJA-Esterase kommt nativ als monomeres 26 kDa großes Protein vor. Unter denaturierenden Bedingungen konnte ein Molekulargewicht von 28 kDa bestimmt werden. Eine Analyse mittels ESI-TOF-Massenspektrometrie ergab ein Molekulargewicht von 28547 Da. Die native MeJA-Esterase hatte ein basisches pH-Optimum von 9.0. Optimale katalytische Aktivität zeigte die MeJA-Esterase bei einer Reaktionstemperatur von 40 C. Der isoelektrische Punkt lag bei pH 4.7. Eine vollständige und irreversible Hemmung der MeJA-Esterase konnte durch 5 mM Phenylmethylsulfonylfluorid (PMSF), einem Serinprotease-Inhibitor erzielt werden. Dieses Ergebnis lieferte einen Hinweis darauf, dass die MeJA-Esterase eine katalytische Triade mit einem reaktiven Serin-Rest besitzt. N-Methylmaleimid, Iodacetamid, Bestatin, Pepstatin und Leupeptin konnten die MeJA-Esterase nicht inhibieren. Nach der Reinigung der MeJA-Esterase wurde das Protein partiell mit der Endoproteinase LysC verdaut. Mittels Sequenzierung der Spaltpeptide und N-terminaler Sequenzierung der MeJA-Esterase konnte von vier Peptiden die Sequenz bestimmt werden. Ein Datenbankvergleich (SwissProt und EMBL) dieser Peptide mit bekannten Sequenzen zeigte eine hohe Homologie (bis zu 80 %) zu verschiedenen Esterasen und α-Hydroxynitrillyasen. Die Peptide konnten somit eindeutig als Bestandteile einer Esterase identifiziert werden. Zur Identifizierung des MeJA-Esterase Gens wurden aus den Peptidsequenzen degenerierte Primer abgeleitet und zur weiteren Klonierung verwendet. Über eine Reverse Transkription mit anschließender PCR wurde ein internes cDNA-Fragment (513 bp) amplifiziert. Mittels RACE (Rapid Amplification of cDNA Ends) konnten das 5´-und 3´-Ende der MeJA-Esterase cDNA ermittelt werden. Die Nucleotidsequenz umfasste einen offenen Leserahmen von 786 bp. Die davon abgeleitete Aminosäuresequenz codierte ein offenes Leseraster für ein Protein von 262 Aminosäuren. Datenbankvergleiche der vollständigen Aminosäuresequenz zeigten Homologien von 33 – 47 % zu Esterasen und α-Hydroxynitrillyasen. Die Aminosäuren der katalytischen Triade, die in den homologen Proteinen hochkonserviert waren, konnten bei der MeJA-Esterase als Serin-83, Asparaginsäure-211 und Histidin-240 ermittelt werden. Diese drei Aminosäuren bilden vermutlich das katalytische Zentrum der MeJA-Esterase. Darüber hinaus konnte eine hochkonservierte Signatur, die allen Lipasen gemeinsam ist in der Aminosäuresequenz der MeJA-Esterase identifziert werden. Diese Ergebnisse erlauben eine Einordnung der MeJA-Esterase in die Superfamilie der „alpha/beta-Fold“-Hydrolasen. Untersuchungen der Primärstruktur der MeJA-Esterase legten den Schluss nahe, dass es sich um ein cytosolisches Enzym handelt. Eine Southern-Blot Analyse mit genomischer DNA aus L. esculentum wurde zur Abschätzung der Kopienzahl der zum Protein der MeJA-Esterase korresporendierenden Gene durchgeführt. Dabei wurden zwei bis sieben DNA-Abschnitte ermittelt, die mit der Volllänge-Sonde der MeJA-Esterase hybridisierten. Dieses Ergebnis lässt vermuten, dass die MeJA-Esterase zu einer Genfamilie gehört. Unklar bleibt jedoch, ob es sich um mehrere homologe Gene handelt, oder ob eine Hybridisierung der Volllänge-Sonde mit Pseudogenen erfolgte. Die heterologe Expression der MeJA-Esterase cDNA wurde erfolgreich durchgeführt. Hierdurch wurde der Beweis erbracht werden, dass die klonierte cDNA tatsächlich für das Gen der MeJA-Esterase codierte. Nach Klonierung der cDNA in den pQE70-Expressionsvektor und Transformation in kompetente E. coli (M15) konnte im Proteinrohextrakt eine spezifische Enzymaktivität von 1.64 pkat/mg detektiert werden. In einem 4-stufigen Reinigungsverfahren wurde das heterolog exprimierte Enzym mit einer Ausbeute von 0.8 % bis zur Homogenität 283-fach angereichert. Untersuchungen zur Substratspezifität zeigten, dass native und heterolog exprimierte MeJA-Esterase Methyljasmonat zu Jasmonsäure hydrolysierten. In beiden Fällen handelte es sich jedoch um kein hochspezifisches Enzym. Für die native MeJA-Esterase konnte ein KM-Wert von 14.7 ± 0.8 µM und für die heterolog exprimierte MeJA-Esterase ein KM-Wert von 24.3 ± 2.3 µM ermittelt werden.From cell suspension cultures of Lycopersicon esculentum a so far unknown enzyme was purified, catalyzing the cleavage of methyl jasmonate to jasmonic acid. The isolated enzyme was termed as methyl jasmonate esterase (MeJA esterase). Qualitative and quantitative enzyme assays using methyl-[2-14C]jasmonic acid and [methyl-3H]methyl jasmonate as substrates were established for purification and characterization of the enzyme. Screening of 18 suspension plant cell cultures of taxonomically distant species revealed that MeJA-Esterase activity occurs in all plant species so far analyzed. MeJA esterase activity was also found in flowers, roots, stems and leaves of L. esculentum (cv. Moneymaker). MeJA esterase was purified in a six-step purification scheme. The enzyme was purified 767-fold to give a homogenous protein with a yield of 2.2 %. The native enzyme exhibited a Mr of 26 kDa (gel-filtration chromatography) which was similar to the Mr determined by SDS-PAGE (Mr of 28 kDa) and ESI-TOF MS analysis (Mr of 28547 kDa). MeJA esterase revealed a pH optimum of pH 9.0 and a temperature optimum of 40 C. Chromatofocussing of MeJA esterase gave an isoelectric point (pI) of 4.7. Phenylmethanesulfonyl fluoride (PMSF), a serine protease inhibitor, led to irreversible and complete inhibition of MeJA esterase at a concentration of 5 mM. This result suggests that the enzyme has a catalytic triade with an active serine residue. N-Methylmaleimide, iodacetamide, bestatin, leupeptin and pepstatin did not inactivate the enzyme. Proteolysis of the purified and pure enzyme with endoproteinase LysC and subsequently sequencing revealed three peptide fragments. N-Terminal sequencing yielded an additional peptide fragment. Sequence alignment of these fragments showed high homologies (up to 80 %) to certain esterases and hydroxynitrile lyases. MeJA esterase peptides could be identified as components of an esterase. For the identification of the MeJA esterase gene degenerated primers were designed on the basis of the partial amino acid sequences obtained from the purified MeJA esterase. Degenerated primers were used for cloning. Reverse transcription followed by PCR amplified an internal cDNA fragment (513 bp). Full-length cDNA of MeJA esterase was amplified using RACE (Rapid amplification of cDNA ends). Sequencing and annealing of the 5´- and the 3´-sequence revealed an open reading frame of 789 bp encoding a 262 amino acid protein. Sequence analysis and alignment with known proteins from Genbank showed high overall-homology of 33 – 47 % to esterases and hydroxynitrile lyases. Since all the aligned proteins belong to the extremely divergent family of alpha/beta-hydrolase fold proteins it could be assumed that MeJA esterase is a member of this protein family. MeJA esterase shows the highly conserved amino acid residues forming the catalytic triad – nucleophile, acid and a histidine – represented by serine-83, aspartic acid-211 and histidine-240. Additionally a higly conserved lipase motive could be identified in the MeJA esterase amino acid sequence. Analysis of the primary structure of the enzyme makes a cytosolic location probably. Southern blot analysis of genomic cDNA from cell suspension cultures of L. esculentum was carried out to estimate gene copies of MeJA esterase. MeJA esterase coding sequence hybridized with two to seven cDNA fragments. It might be possible that MeJA esterase belongs to a gene family. It remains still unclear if there are several homologous genes or pseudogenes hybridized with the MeJA esterase sequence. To get unequivocal evidence of the identity of cloned sequence, MeJA esterase cDNA was successfully subcloned into a bacterial vector (pQE70) for heterologous expression. Crude extracts of E. coli M15 cells harbouring the pQE-MeJA esterase plasmid showed MeJA esterase activity (1.64 pkat/mg) while wild type M15 cells did not show any MeJA esterase activity. A four step purification protocol was employed to purify the enzyme to homogeneity (283-fold) with a yield of 0.8 %. Analysis of substrate specifity showed that native and heterologously expressed MeJA esterase cleaved methyl jasmonate to jasmonic acid. However MeJA esterase is not a highly specific enzyme. Enzyme kinetics of native MeJA esterase revealed a KM value of 14.7 ± 0.8 µM while heterologous expressed MeJA esterase revealed a KM value of 24.3 ± 2.3 µM. Northern blot analysis was used to determine MeJA esterase expression in different plant organs. MeJA esterase transcripts were present in all tomato plant tissues. High levels of MeJA esterase mRNA could be found in roots and flowers while low to moderate amounts where present in leafs and stems of tomato plants

Pride, plague & protest : a comparative study of defiance in AIDS campaigns, testimonies and protest art by affected gay and Black African men

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Therapy of children with septo-optic dysplasia (SOD)- experiences in the ophthalmic clininc and the paediatric clinic of the University Erlangen-Nuernberg

Die septo-optische Dysplasie (SOD) ist eine seltene Erkrankung bislang ungeklärter Genese mit starker phänotypischer Variabilität. Sie wird zu den sog. Mittelliniendefekten gezählt. Neben ophthalmologischen und neuroendokrinologischen Zeichen und Symptomen gehört auch ein fehlendes Septum pellucidum zum Befundkomplex. Ophthamologisch liegen eine Mikropapille mit Sehnervhypoplasie und Nystagmen vor. Die neuroendokrinologischen Ausfälle betreffen die hypothalamo-hypophysäre Hormonachse. Unser Patientenkollektiv setzte sich aus insgesamt sechs Mädchen und vier Jungen zusammen, die zwischen 1992 und 2006 in der Kinder- und Augenklinik behandelt wurden. Anhand der Daten wurden der Diagnosezeitpunkt, der Verlauf der Erkrankung und durchgeführte Untersuchungen, die Bildgebung, ophthalmologische (Re-) Evaluation und neuroendokrinologische Funktionsdiagnostik umfassten, untersucht. Die Verdachtsdiagnose SOD und die Sicherung dieser Diagnose können bereits frühzeitig über die ophthalmologische Untersuchung erhoben werden. Daher empfiehlt es sich, bei okulären Auffälligkeiten (z.B. Mikropapille) im Neugeborenen- und Säuglingsalter eine SOD zu erwägen und entsprechend weiterführende neuroendokrinologische Aufklärung einzuleiten.Septo-optic dysplasia (SOD) is a rare disease of so far unknown aetiology and high phenotypical variability. It is part of the so-called midline defects. Neuroendocrinically and ophthalmologic symptoms as well as the agenesia of septum pellucidum belong to the diagnostic findings. Ophthalmologic diagnostics include hypoplasia of the optic nerve with micropapilla and nystagmus. Our patient population consisted of six girls and four boys, all examined between 1992-2006 in our paediatric and ophthalmic clinic. According to the records there were evaluated the date of diagnosis, the progress of the disease and examinations, like radiological staging, ophthalmologic (re-)evaluation and neuro-endocrinically testing. The suspicion of SOD and its confirmation can be earlier diagnosed by further evaluation of ophthalmologic abnormalities. Therefore it makes sense to think of this option in the case of ocular findings (e.g. micropapilla) in the newborn and infant period in order to add neuro-endocrinically examination

Phytochemistry

Screening of 18 suspension plant cell cultures of taxonomically distant species revealed that a methyl jasmonate hydrolysing enzyme activity (0.21-5.67 pkat/mg) occurs in all species so far analysed. The methyl jasmonate hydrolysing esterase was purified from cell cultures of Lycopersicon esculentum using a five-step procedure including anion-exchange chromatography, gel-filtration and chromatography on hydroxylapatite. The esterase was purified 767-fold to give an almost homogenous protein in a yield of 2.2%. The native enzyme exhibited a M-r of 26 kDa (gel-filtration chromatography), which was similar to the M-r determined by SDS-PAGE and MALDI-TOF analysis (M-r of 28547 kDa). Enzyme kinetics revealed a K-m value of 15 muM and a V-max value of 7.97 nkat/mg, an pH optimum of 9.0 and a temperature optimum of 40 degreesC. The enzyme also efficiently hydrolyzed methyl esters of abscisic acid, indole-3-acetic acid, and fatty acids. In contrast, methyl esters of salicylic acid, benzoic acid and cinnamic acid were only poor substrates for the enzyme. N-Methylmaleimide, iodacetamide, bestatin and pepstatin (inhibitors of thiol-, metal- and carboxyproteases, respectively) did not inactivate the enzyme while a serine protease inhibitor, phenylmethylsulfonyl fluoride, at a concentration of 5 mM led to irreversible and complete inhibition of enzyme activity. Proteolysis of the pure enzyme with endoproteinase LysC revealed three peptide fragments with 11-14 amino acids. N-Terminal sequencing yielded an additional peptide fragment with 10 amino acids. Sequence alignment of these fragments showed high homologies to certain plant esterases and hydroxynitrile lyases that belong to the alpha/beta hydrolase fold protein superfamily. (C) 2002 Published by Elsevier Science Ltd

Cloning and expression of a tomato cDNA encoding a methyl jasmonate cleaving esterase.

Jasmonic acid and its methyl ester are ubiquitous plant signalling compounds necessary for the regulation of growth and development, as well as for the response of plants to environmental stress factors. To date, it is not clear whether methyl jasmonate itself acts as a signal or if its conversion to jasmonic acid is mandatory prior to the induction of a defense response. We have cloned a cDNA, encoding a methyl jasmonate-cleaving enzyme, from tomato cell suspension cultures. Sequence analysis revealed significant similarity to plant esterases and to (S)-hydroxynitrile lyases with an alpha/beta-hydrolase fold structure. The coding sequence was heterologously expressed in Escherichia coli and purified in a catalytically active form. Transcript levels, as well as enzymatic activity, were determined in different tomato tissues. High transcript levels and enzyme activities were found in roots and flowers, while the mRNA level and activity were low in stems and leaves. Moreover, when tested in methyl jasmonate- and elicitor-treated cell suspension cultures, transcript levels were found to decrease, indicating that this particular enzyme might be a regulator of jasmonate signalling

Completion of FLOT Therapy, Regardless of Tumor Regression, Significantly Improves Overall Survival in Patients with Esophageal Adenocarcinoma

Esophageal cancer is the eighth most common cancer worldwide, with poor prognosis and high mortality. The combination of surgery and systemic therapy provide the best chances for long-term survival. The purpose of this study was to analyze the impact of the FLOT protocol on the overall survival of patients following surgery for esophageal adenocarcinoma, with a focus on the patients who did not benefit in terms of pathological remission from the neoadjuvant therapy. A retrospective analysis of all the patients who underwent esophagectomies from 2012 to 2017 for locally advanced adenocarcinomas of the esophagus at a tertiary medical center was performed. The results show that the completion of systemic therapy, regardless of the tumor regression grading, had a significant positive impact on the overall survival. The patients with complete regression and complete systemic therapy showed the best outcomes. Anastomotic insufficiency did not negatively impact the long-term survival, while complications of the systemic therapy led to significantly reduced overall survival. We conclude that adjuvant systemic therapy should, when possible, always be completed, regardless of the tumor regression, following an esophagectomy