LIMB REPLANTATION AFTER AVULSION INJURIES: TECHNIQUES AND TACTICS FOR SUCCESS

Objectives: Retrospective evaluation of cases of limb replantation after avulsion injuries. Evaluation of the techniques and tactics used, that contributed to success and good functional results. Methods: Forty-three patients' records were assessed. All the cases had been submitted to limb replantation after avulsion injuries. Results: The majority of the cases were young men. The most common injury was to the thumbs. The surgical techniques and tactics used were: nerve grafting, vein grafting, transposition of the digital vessels, limb shortening, and heterotopic replantation. The most commonly used technique was vein graft. The limb survival rate was high (93%), as was patient satisfaction. Conclusion: Replantation after avulsion injury depends on the correct diagnosis of the limb viability and the use of appropriate surgical techniques and tactics for each case. The experience of the team of surgeons and a good hospital structure are essential for good results. There are few articles in medical literature about the indications, techniques and results of limb replantation after avulsion injuries. We believe that this retrospective evaluation can bring new information and contributions to the correct management of this highly complex situation. Level of evidence IV, Case Series.

An extraordinary new species of Melanophryniscus (Anura, Bufonidae) from southeastern Brazil

We describe a new species of bufonid from a lowland, sandy soil, restinga habitat in the state of Espirito Santo, southeastern Brazil. Based on the shared occurrence of putative morphological synapomorphies of Melanophryniscus and the results of a phylogenetic analysis of DNA sequences of a broad sample of bufonids, and other anurans, we assign the new species to Melanophryniscus. The new species possesses several peculiar character states that distinguish it from all other Melanophryniscus including, but not limited to: fingers II, III, and V much reduced; nuptial pad with few enlarged, brown-colored spines on medial margin of finger II; seven presacral vertebrae, the last fused with the sacrum; and ventral humeral crest prominent, forming a spinelike projection.

Analysis of MLL2 gene in the first Brazilian family with Kabuki syndrome

Most patients with Kabuki syndrome (KS) are the only person in their family with the condition. However, familial cases of KS have been described showing evidence that this syndrome can be inherited as a dominant trait with variable expressivity. We report on two related individuals with facial findings characteristic of KS. The proposita had arched eyebrows, long and upward slanting palpebral fissures, cleft lip and palate, retromicrognathia, brachydactyly of hands and feet, stubby fingers, nail hypoplasia, and prominent finger pads. Her mother had eyebrows with dispersed lateral half, long and upward slanting palpebral fissures, retrognathia, abnormal and posteriorly rotated ears, prominent finger pads, brachydactyly of feet, learning difficulties, and psychomotor development delay. DNA sequencing revealed a novel missense mutation in the MLL2 gene in both the proposita and her mother. The mutation (p.R5432Q) was found in the exon 51, within the SET domain of the gene, which confers methyltransferase activity on the protein. Therefore, the epigenetic and transcriptional regulatory properties of this protein may be altered and this suggests that the mutation is the cause of phenotype observed in both the patient and her mother. The clinical signs and the molecular evidence in this family further support the notion that KS is an autosomal dominant condition with variable expressivity. To our knowledge this is the first report of a Brazilian family with recurrence of this syndrome. (C) 2012 Wiley Periodicals, Inc.

A relict new species of Oreobates (Anura, Strabomantidae) from the Seasonally Dry Tropical Forests of Minas Gerais, Brazil, and its implication to the biogeography of the genus and that of South American Dry Forests

A new species of Oreobates is described from Cavernas do Peruacu National Park, Januaria, Minas Gerais state, in the Atlantic Dry Forests of Brazil. The new species is distinguished from all other Oreobates by having the following combination of characters: large tympanum, discs broadly enlarged and truncate on Fingers III and IV, smooth dorsal skin, nuptial pads absent, snout subacuminate, and a very short pulsatile (2-3 pulses) single-noted advertisement call with dominant frequency of about 3150 Hz, and no harmonic structure. Molecular phylogenetic analyses using partial sequences of the mitochondrial genes cytochrome b (cyt b) and 16S using multiple outgroups recovered the new species within Oreobates and sister to O. heterodactylus. The latter species inhabits the Dry Forests of Mato Grosso (Cerrado) and Bolivia (Chiquitano forests), and is strictly associated to these habitats, which suggests a preterit connection between Chiquitano and Atlantic Dry Forests. The discovery of a new Oreobates in the Atlantic Dry Forest is of great importance for the conservation of these dry forests, as it is known only from this type of habitat.

Mechanism of resistance to tyrosine kinase inhibitors in philadelphia-positive acute lymphblastic leukaemia (all): from genetic alterations to impaired RNA editing

The Ph chromosome is the most frequent cytogenetic aberration associated with adult ALL and it represents the single most significant adverse prognostic marker. Despite imatinib has led to significant improvements in the treatment of patients with Ph+ ALL, in the majority of cases resistance developed quickly and disease progressed. Some mechanisms of resistance have been widely described but the full knowledge of contributing factors, driving both the disease and resistance, remains to be defined. The observation of rapid development of lymphoblastic leukemia in mice expressing altered Ikaros (Ik) isoforms represented the background of this study. Ikaros is a zinc finger transcription factor required for normal hemopoietic differentiation and proliferation, particularly in the lymphoid lineages. By means of alternative splicing, Ikaros encodes several proteins that differ in their abilities to bind to a consensus DNA-binding site. Shorter, DNA nonbinding isoforms exert a dominant negative effect, inhibiting the ability of longer heterodimer partners to bind DNA. The differential expression pattern of Ik isoforms in Ph+ ALL patients was analyzed in order to determine if molecular abnormalities involving the Ik gene could associate with resistance to imatinib and dasatinib. Bone marrow and peripheral blood samples from 46 adult patients (median age 55 yrs, 18-76) with Ph+ ALL at diagnosis and during treatment with imatinib (16 pts) or dasatinib (30 pts) were collected. We set up a fast, high-throughput method based on capillary electrophoresis technology to detect and quantify splice variants. 41% Ph+ ALL patients expressed high levels of the non DNA-binding dominant negative Ik6 isoform lacking critical N-terminal zinc-fingers which display abnormal subcellular compartmentalization pattern. Nuclear extracts from patients expressed Ik6 failed to bind DNA in mobility shift assay using a DNA probe containing an Ikaros-specific DNA binding sequence. In 59% Ph+ ALL patients there was the coexistence in the same PCR sample and at the same time of many splice variants corresponded to Ik1, Ik2, Ik4, Ik4A, Ik5A, Ik6, Ik6 and Ik8 isoforms. In these patients aberrant full-length Ikaros isoforms in Ph+ ALL characterized by a 60-bp insertion immediately downstream of exon 3 and a recurring 30-bp in-frame deletion at the end of exon 7 involving most frequently the Ik2, Ik4 isoforms were also identified. Both the insertion and deletion were due to the selection of alternative splice donor and acceptor sites. The molecular monitoring of minimal residual disease showed for the first time in vivo that the Ik6 expression strongly correlated with the BCR-ABL transcript levels suggesting that this alteration could depend on the Bcr-Abl activity. Patient-derived leukaemia cells expressed dominant-negative Ik6 at diagnosis and at the time of relapse, but never during remission. In order to mechanistically demonstrated whether in vitro the overexpression of Ik6 impairs the response to tyrosine kinase inhibitors (TKIs) and contributes to resistance, an imatinib-sensitive Ik6-negative Ph+ ALL cell line (SUP-B15) was transfected with the complete Ik6 DNA coding sequence. The expression of Ik6 strongly increased proliferation and inhibited apoptosis in TKI sensitive cells establishing a previously unknown link between specific molecular defects that involve the Ikaros gene and the resistance to TKIs in Ph+ ALL patients. Amplification and genomic sequence analysis of the exon splice junction regions showed the presence of 2 single nucleotide polymorphisms (SNPs): rs10251980 [A/G] in the exon2/3 splice junction and of rs10262731 [A/G] in the exon 7/8 splice junction in 50% and 36% of patients, respectively. A variant of the rs11329346 [-/C], in 16% of patients was also found. Other two different single nucleotide substitutions not recognized as SNP were observed. Some mutations were predicted by computational analyses (RESCUE approach) to alter cis-splicing elements. In conclusion, these findings demonstrated that the post-transcriptional regulation of alternative splicing of Ikaros gene is defective in the majority of Ph+ ALL patients treated with TKIs. The overexpression of Ik6 blocking B-cell differentiation could contribute to resistance opening a time frame, during which leukaemia cells acquire secondary transforming events that confer definitive resistance to imatinib and dasatinib.

Aldo Rossi, autobiografia scientifica: scrittura come progetto. Indagine critica tra scrittura e progetto di architettura

«In altri termini mi sfuggiva e ancora oggi mi sfugge gran parte del significato dell’evoluzione del tempo; come se il tempo fosse una materia che osservo dall’esterno. Questa mancanza di evoluzione è fonte di alcune mie sventure ma anche mi appartiene con gioia.» Aldo Rossi, Autobiografia scientifica. The temporal dimension underpinning the draft of Autobiografia scientifica by Aldo Rossi may be referred to what Lucien Lévy-Bruhl, the well-known French anthropologist, defines as “primitive mentality” and “prelogical” conscience : the book of life has lost its page numbers, even punctuation. For Lévy-Bruhl, but certainly for Rossi, life or its summing up becomes a continuous account of ellipses, gaps, repetitions that may be read from left to right or viceversa, from head to foot or viceversa without distinction. Rossi’s autobiographical writing seems to accept and support the confusion with which memories have been collected, recording them after the order memory gives them in the mental distillation or simply according to the chronological order in which they have happened. For Rossi, the confusion reflects the melting of memory elements into a composite image which is the result of a fusion. He is aware that the same sap pervades all memories he is going to put in order: each of them has got a common denominator. Differences have diminished, almost faded; the quick glance is prevalent over the distinction of each episode. Rossi’s writing is beyond the categories dependent on time: past and present, before and now. For Rossi, the only repetition – the repetition the text will make possible for an indefinite number of times – gives peculiarity to the event. As Gilles Deleuze knows, “things” may only last as “singleness”: more frequent the repetition is, more singular is the memory phenomenon that recurs, because only what is singular magnifies itself and happens endlessly forever. Rossi understands that “to raise the first time to nth forever”, repetition becomes glorification . It may be an autobiography that, celebrating the originality, enhances the memory event in the repetition; in fact it greatly differs from the biographical reproduction, in which each repetition is but a weaker echo, a duller copy, provided with a smaller an smaller power in comparison with the original. Paradoxically, for Deleuze the repetition asserts the originality and singularity of what is repeated. Rossi seems to share the thought expressed by Kierkegaard in the essay Repetition: «The hope is a graceful maiden slipping through your fingers; the memory of an elderly woman, indeed pretty, but never satisfactory if necessary; the repetition is a loved friend you are never tired of, as it is only the new to make you bored. The old never bores you and its presence makes you happy [...] life is but a repetition [...] here is the beauty of life» . Rossi knows well that repetition hints at the lasting stability of cosmic time. Kierkegaard goes on: «The world exists, and it exists as a repetition» . Rossi devotes himself, on purpose and in all conscience, to collect, to inventory and «to review life», his own life, according to a recovery not from the past but of the past: a search work, the «recherche du temps perdu», as Proust entitled his masterpiece on memory. If you want the past time to be not wasted, you must give it presence. «Memoria e specifico come caratteristiche per riconoscere se stesso e ciò che è estraneo mi sembravano le più chiare condizioni e spiegazioni della realtà. Non esiste uno specifico senza memoria, e una memoria che non provenga da un momento specifico; e solo questa unione permette la conoscenza della propria individualità e del contrario (self e non-self)» . Rossi wants to understand himself, his own character; it is really his own character that requires to be understood, to increase its own introspective ability and intelligence. «Può sembrare strano che Planck e Dante associno la loro ricerca scientifica e autobiografica con la morte; una morte che è in qualche modo continuazione di energia. In realtà, in ogni artista o tecnico, il principio della continuazione dell’energia si mescola con la ricerca della felicità e della morte» . The eschatological incipit of Rossi’s autobiography refers to Freud’s thought in the exact circularity of Dante’s framework and in as much exact circularity of the statement of the principle of the conservation of energy: in fact it was Freud to connect repetition to death. For Freud, the desire of repetition is an instinct rooted in biology. The primary aim of such an instinct would be to restore a previous condition, so that the repeated history represents a part of the past (even if concealed) and, relieving the removal, reduces anguish and tension. So, Freud ask himself, what is the most remote state to which the instinct, through the repetition, wants to go back? It is a pre-vital condition, inorganic of the pure entropy, a not-to-be condition in which doesn’t exist any tension; in other words, Death. Rossi, with the theme of death, introduces the theme of circularity which further on refers to the sense of continuity in transformation or, in the opposite way, the transformation in continuity. «[...] la descrizione e il rilievo delle forme antiche permettevano una continuità altrimenti irripetibile, permettevano anche una trasformazione, una volta che la vita fosse fermata in forme precise» . Rossi’s attitude seems to hint at the reflection on time and – in a broad sense – at the thought on life and things expressed by T.S. Eliot in Four Quartets: «Time present and time past / Are both perhaps present in time future, / And time future is contained in time past. / I all time is eternally present / All time is unredeemable. / What might have been is an abstraction / Remaining perpetual possibility / Only in a word of speculation. / What might have been and what has been / Point to one end, which is always present. [...]» . Aldo Rossi’s autobiographical story coincides with the description of “things” and the description of himself through the things in the exact parallel with craft or art. He seems to get all things made by man to coincide with the personal or artistic story, with the consequent immediate necessity of formulating a new interpretation: the flow of things has never met a total stop; all that exists nowadays is but a repetition or a variant of something existing some time ago and so on, without any interruption until the early dawnings of human life. Nevertheless, Rossi must operate specific subdivisions inside the continuous connection in time – of his time – even if limited by a present beginning and end of his own existence. This artist, as an “historian” of himself and his own life – as an auto-biographer – enjoys the privilege to be able to decide if and how to operate the cutting in a certain point rather than in another one, without being compelled to justify his choice. In this sense, his story is a matter very ductile and flexible: a good story-teller can choose any moment to start a certain sequence of events. Yet, Rossi is aware that, beyond the mere narration, there is the problem to identify in history - his own personal story – those flakings where a clean cut enables the separation of events of different nature. In order to do it, he has to make not only an inventory of his own “things”, but also to appeal to authority of the Divina Commedia started by Dante when he was 30. «A trent’anni si deve compiere o iniziare qualcosa di definitivo e fare i conti con la propria formazione» . For Rossi, the poet performs his authority not only in the text, but also in his will of setting out on a mystical journey and handing it down through an exact descriptive will. Rossi turns not only to the authority of poetry, but also evokes the authority of science with Max Plank and his Scientific Autobiography, published, in Italian translation, by Einaudi, 1956. Concerning Planck, Rossi resumes an element seemingly secondary in hit account where the German physicist «[...] risale alle scoperte della fisica moderna ritrovando l’impressione che gli fece l’enunciazione del principio di conservazione dell’energia; [...]» . It is again the act of describing that links Rossi to Planck, it is the description of a circularity, the one of conservation of energy, which endorses Rossi’s autobiographical speech looking for both happiness and death. Rossi seems to agree perfectly to the thought of Planck at the opening of his own autobiography: «The decision to devote myself to science was a direct consequence of a discovery which was never ceased to arouse my enthusiasm since my early youth: the laws of human thought coincide with the ones governing the sequences of the impressions we receive from the world surrounding us, so that the mere logic can enable us to penetrate into the latter one’s mechanism. It is essential that the outer world is something independent of man, something absolute. The search of the laws dealing with this absolute seems to me the highest scientific aim in life» . For Rossi the survey of his own life represents a way to change the events into experiences, to concentrate the emotion and group them in meaningful plots: «It seems, as one becomes older. / That the past has another pattern, and ceases to be a mere sequence [...]» Eliot wrote in Four Quartet, which are a meditation on time, old age and memory . And he goes on: «We had the experience but missed the meaning, / And approach to the meaning restores the experience / In a different form, beyond any meaning [...]» . Rossi restores in his autobiography – but not only in it – the most ancient sense of memory, aware that for at least 15 centuries the Latin word memoria was used to show the activity of bringing back images to mind: the psychology of memory, which starts with Aristotele (De Anima), used to consider such a faculty totally essential to mind. Keith Basso writes: «The thought materializes in the form of “images”» . Rossi knows well – as Aristotele said – that if you do not have a collection of mental images to remember – imagination – there is no thought at all. According to this psychological tradition, what today we conventionally call “memory” is but a way of imagining created by time. Rossi, entering consciously this stream of thought, passing through the Renaissance ars memoriae to reach us gives a great importance to the word and assumes it as a real place, much more than a recollection, even more than a production and an emotional elaboration of images.

The study of a novel zinc finger gene cluster TZF and a genomic region flanking the histone H 4 replacement gene H 4 r of Drosophila melanogaster

Part I : A zinc finger gene Tzf1 was cloned in the earlier work of the lab by screening a ë-DASH2 cDNA expression library with an anti-Rat SC antibody. A ë-DASH2 genomic DNA library and cosmid lawrist 4 genomic DNA library were screened with the cDNA fragment of Tzf1 to determine the genomic organization of Tzf1. Another putative zinc finger gene Tzf2 was found about 700 bp upstream of Tzf1.RACE experiment was carried out for both genes to establish the whole length cDNA. The cDNA sequences of Tzf and Tzf2 were used to search the Flybase (Version Nov, 2000). They correspond to two genes found in the Flybase, CG4413 and CG4936. The CG4413 transcript seems to be a splicing variant of Tzf transcripts. Another two zinc finger genes Tzf3 and Tzf4 were discovered in silico. They are located 300 bp away from Tzf and Tzf2, and a non-tandem cluster was formed by the four genes. All four genes encode proteins with a very similar modular structure, since they all have five C2H2 type zinc fingers at their c-terminal ends. This is the most compact zinc finger protein gene cluster found in Drosophila melanogaster.Part II: 34,056 bp insert of the cosmid 19G11

Regulation und Funktion von hunchback in Neuroblasten-Zellstammbäumen von Drosophila melanogaster

In Drosophila melanogaster wird das Nervensystem von neuralen Vorläuferzellen, den Neuroblasten (NB) gebildet. Diese teilen sich im Stammzellmodus und bringen bei jeder Teilung eine Ganglienmutterzelle (GMZ) hervor. GMZ teilen sich einmal und generieren zwei Zellen, die ein neuronales und/oder gliales Schicksal annehmen. Die Reihenfolge in der ein NB GMZ generiert, ist vermutlich ein zellautonomer Prozess, der an die transiente und sequenzielle Expression der Transkriptionsfaktoren Hunchback (Hb), Krüppel (Kr), Pdm, Castor (Cas) und Grainy head (Grh) gekoppelt ist. Hb ist der erste Faktor dieser Genkaskade. Damit der NB zur nächsten zeitlichen Identität übergehen kann, die in der Regel von Kr bestimmt wird, muss die Hb-Expression im NB beendet werden. Das Abschalten wird transkriptionell reguliert und ist von einer vorhergehenden Mitose abhängig. Im Gegensatz zum NB wird in der GMZ, die bei dieser Teilung entsteht, die Hb-Expression beibehalten. In der vorliegenden Arbeit wurde der Mitose-abhängige Mechanismus aufgeklärt, durch den eine selektive Abschaltung von hb im NB erfolgt, nicht aber in der GMZ. Der Transkriptionsfaktor Seven-up (Svp) beendet die hb-Expression im NB. svp wird vor der entscheidenden Zellteilung im NB transkribiert, aber kaum translatiert. Nach erfolgter Zellteilung wird Svp verstärkt translatiert und sowohl im NB als auch in der neu entstandenen GMZ exprimiert. Im NB führt die Svp-Funktion zur Abschaltung von Hb. In der ebenfalls Svp-positiven GMZ verhindert jedoch Prospero (Pros), das während der Teilung in diese Zelle segregiert ist, die Aktivität von Svp. Um zu überprüfen, ob Svp und Pros die Transkription von Hb entweder direkt oder indirekt regulieren, wurde eine Enhancerfragment-Analyse durchgeführt. Durch diese Unter-suchungen sollten relevante regulatorische Bereiche des hb-Gens identifiziert werden. Zusätzlich zu den bereits bekannten Regionen im 5´-Bereich von hb, konnten 3´ des transkribierten Bereiches weitere nervensystemspezifische Enhancerelemente gefunden werden. Neben der Spezifizierung von früh geborenen Nachkommen von NB hat Hb im Nervensystem die Funktion, den multipotenten Zustand junger NB aufrecht zu erhalten. Um den Beitrag einzelner Domänen des Hb-Proteins an diesen Aufgaben zu bestimmen, wurden verschiedene hb-Varianten, in denen spezifische Domänen deletiert bzw. mutiert waren, im NB7-1 überexprimiert und auf die Induktion von „frühem“ Schicksal getestet. In einem zweiten Ansatz wurden mutante hb-Allele analysiert. Es stellte sich heraus, dass der D-Box eine entscheidende Rolle bei der Spezifizierung von „frühem“ Schicksal zukommt. Die terminalen Zinkfinger sind vermutlich in die Aufrechterhaltung der Kompetenz von NB involviert. Eine Deletion der A-Box erhöht möglicherweise die Aktivität des Hb-Proteins.

Ansatz zur Generierung einer konditionalen, reversiblen Wt1 k.o.-Maus

Ansatz zur Generierung einer konditionalen, reversiblen Wt1 k.o.-Maus Der Wilms-Tumor (WT, Nephroblastom) ist ein embryonaler Nierentumor, der durch die maligne Transformation von undifferenziertem Nierengewebe, sog. nephrogenen Resten, entsteht. WT treten mit einer Inzidenz von 1 in 10.000 Lebendgeburten auf. Das Hauptmanifestationsalter, der normalerweise einseitig und sporadisch auftretenden Tumore, liegt zwischen dem 3. und 4. Lebensjahr. Etwa 10 % der Patienten entwickeln jedoch bilaterale Tumore. In diesen Fällen ist eine Assoziation mit komplexen genetischen Krankheitsbildern (u. a. WAGR-, Denys-Drash-, Frasier- und Beckwith-Wiedemann-Syndrom) festzustellen. In 15 % der sporadischen WT sind Mutationen im WT1 (Wilms-Tumor 1)-Gen beschrieben. WT1 besteht aus zehn Exons und weist typische Merkmale von Transkriptionsfaktoren (z. B. vier Zinkfinger) auf. Zwei alternative Spleißereignisse betreffen Exon 5 (+/−Exon 5) und Exon 9 (Transkripte mit bzw. ohne die codierenden Sequenzen für die AS Lysin-Threonin-Serin; +/−KTS). Die Lage der drei alternativ vorhandenen AS zwischen den Zinkfingern 3 und 4 bestimmt die verschiedenen Funktionen der WT1-Proteine (4 Isoformen) als Transkriptionsfaktor (−KTS) bzw. als RNA-bindendes Protein (+KTS). Das zunächst im Zusammenhang mit WT als Tumorsuppressorgen identifizierte WT1 ist ein Entwicklungsgen mit einem sehr komplexen Expressionsmuster in der Embryonalentwicklung. Dabei ist v. a. die Bedeutung in der Urogenitalentwicklung entscheidend. Konstitutive, homozygote Wt1−/− k.o.-Mäuse sind embryonal (~ E12,5 dpc) letal und bilden u. a. keine Gonaden und keine Nieren. Aus diesem Grund existiert bisher kein Wilms-Tumormodell. Die Herstellung eines konditionalen murinen Tiermodells auf Basis des Tet on/off-Systems zur Untersuchung der Nierenentwicklung bzw. zur Analyse der Wilms-Tumorpathogenese war Ziel dieser Arbeit. Hierfür wurden drei Mauslinien generiert: Zwei transgene sog. Responder-Linien, die eine chimäre spleißbare Wt1-cDNA der Variante musWt1+Exon 5;+/−KTS unter der Kontrolle eines Tet-responsiven Promotors im Genom tragen. Dieses tTA/Dox-abhängig regulierbare Wt1-Transgen (tgWt1) sollte (exogen regulierbar) die Expression des endogenen Wt1-Lokus ausreichend nachahmen, um die kritischen Phasen der Embryogenese zu überwinden und lebensfähige Tiere zu erhalten. Parallel dazu wurde die Wt1-Effektor-Mauslinie (WE2) generiert. Diese trägt einen tetrazyklinabhängigen Transaktivator (tTA) zur Steuerung Tet-regulierbarer Transgene unter der Kontrolle des endogenen Wt1-Promotors. Die durch homologe Rekombination in ES-Zellen erreichte Integration des tTA direkt am Translationsstartpunkt des Wt1-Lokus hat in den Tieren einen heterozygoten Wt1 knock out/tTA knock in zur Folge. Die bisher vorgenommenen Verpaarungen doppelt transgener Wt1-tTA+/−/Resp-Mäuse ergaben keinen Rescue des letalen Wt1 k.o. und es konnten bislang keine Wilms-Tumore induziert werden. Alle im Verlauf der Arbeit generierten Mauslinien wurden umfassend charakterisiert. So konnte für die Tiere der Responder-Linien Wt1-Resp1 (mit zusätzlichen Isolator-Sequenzen zum Schutz des Transgens vor Positionseffekten) und Wt1-Resp2 (ohne Isolatoren) konnte die Tet-induzierbare Expression und die Spleißbarkeit des tgWt1 in MEF-Assays und mittels Effektor-Mäusen auf RNA-Ebene nachgewiesen werden. Die genomische Charakterisierung der WE2-Linie ergab eine ungeklärte etwa 120 kb große Inversion am Wt1-Lokus, die alle 5'-regulatorischen Sequenzen mitsamt des tTA vom Rest von Wt1 trennt. Tiere dieser Linie weisen aber dennoch einen funktionalen Wt1 k.o. auf: Unter den Nachkommen aus Intercross-Verpaarungen von Wt1-tTA+/−-Mäusen lassen sich auf Grund der Letalität keine Wt1−/−-Genotypen nachweisen. Die Charakterisierung der Effektor-Linie auf RNA-Ebene und mittels Reporter-Mäusen liefert ein Wt1-analoges tTA-Expressionsmuster: So findet man eine deutliche tTA-Expression u. a. in Niere (Glomeruli), Uterus, Ovar und Testis. Die hier vorgestellten Experimente ergeben darüber hinaus eindeutige Hinweise einer Beteiligung von Wt1 in der Entstehung der glatten Muskulatur bzw. in der Vaskulogenese.

Utilizzo dell'esame dermoscopico nella valutazione clinica e nel follow up dei nevi melanocitici in sede acrale dell'età pediatrica: studio retrospettivo osservazionale

La dermoscopia, metodica non invasiva, di pratico utilizzo e a basso costo si è affermata negli ultimi anni come valido strumento per la diagnosi e il follow up delle lesioni cutanee pigmentate e non pigmentate. La presente ricerca è stata incentrata sullo studio dermoscopico dei nevi melanocitici a localizzazione palmo-plantare, acquisiti e congeniti, in età pediatrica: a questo scopo sono state analizzate le immagini dei nevi melanocitici acrali nei pazienti visitati c/o l’ambulatorio di Dermatologia Pediatrica del Policlinico Sant’Orsola Malpighi dal 2004 al 2011 per definire i principali pattern dermoscopici rilevati ed i cambiamenti osservati durante il follow up videodermatoscopico. Nella nostra casistica di immagini dermoscopiche pediatriche abbiamo notato un cambiamento rilevante (inteso come ogni modificazione rilevata tra il pattern demoscopico osservato al baseline e i successivi follow up) nell’88,6% dei pazienti ed in particolare abbiamo osservato come in un’alta percentuale di pazienti (80%), si sia verificato un vero e proprio impallidimento del nevo melanocitico e in un paziente è stata evidenziata totale regressione dopo un periodo di tempo di 36 mesi. E’ stato interessante notare come l’impallidimento della lesione melanocitaria si sia verificata per lo più in sedi sottoposte ad una sollecitazione meccanica cronica, come la pianta del piede e le dita (di mani e piedi), facendoci ipotizzare un ruolo del traumatismo cronico nelle modificazioni che avvengono nelle neoformazioni melanocitarie dei bambini in questa sede.

Synthesis of Hand Exoskeletons for the Rehabilitation of Post-Stroke Patients

This dissertation presents the synthesis of a hand exoskeleton (HE) for the rehabilitation of post-stroke patients. Through the analysis of state-of-the-art, a topological classification was proposed. Based on the proposed classification principles, the rehabilitation HEs were systematically analyzed and classified. This classification is helpful to both understand the reason of proposing certain solutions for specific applications and provide some useful guidelines for the design of a new HE, that was actually the primary motivation of this study. Further to this classification, a novel rehabilitation HE was designed to support patients in cylindrical shape grasping tasks with the aim of recovering the basic functions of manipulation. The proposed device comprises five planar mechanisms, one per finger, globally actuated by two electric motors. Indeed, the thumb flexion/extension movement is controlled by one actuator whereas a second actuator is devoted to the control of the flexion/extension of the other four fingers. By focusing on the single finger mechanism, intended as the basic model of the targeted HE, the feasibility study of three different 1 DOF mechanisms are analyzed: a 6-link mechanism, that is connected to the human finger only at its tip, an 8-link and a 12-link mechanisms where phalanges and articulations are part of the kinematic chain. The advantages and drawbacks of each mechanism are deeply analyzed with respect to targeted requirements: the 12-link mechanism was selected as the most suitable solution. The dimensional synthesis based on the Burmester theory as well as kinematic and static analyses were separately done for all fingers in order to satisfy the desired specifications. The HE was finally designed and a prototype was built. The experimental results of the first tests are promising and demonstrate the potential for clinical applications of the proposed device in robot-assisted training of the human hand for grasping functions.

Design and Characterization of Curved and Spherical Flexure Hinges for Planar and Spatial Compliant Mechanisms

A flexure hinge is a flexible connector that can provide a limited rotational motion between two rigid parts by means of material deformation. These connectors can be used to substitute traditional kinematic pairs (like bearing couplings) in rigid-body mechanisms. When compared to their rigid-body counterpart, flexure hinges are characterized by reduced weight, absence of backlash and friction, part-count reduction, but restricted range of motion. There are several types of flexure hinges in the literature that have been studied and characterized for different applications. In our study, we have introduced new types of flexures with curved structures i.e. circularly curved-beam flexures and spherical flexures. These flexures have been utilized for both planar applications (e.g. articulated robotic fingers) and spatial applications (e.g. spherical compliant mechanisms). We have derived closed-form compliance equations for both circularly curved-beam flexures and spherical flexures. Each element of the spatial compliance matrix is analytically computed as a function of hinge dimensions and employed material. The theoretical model is then validated by comparing analytical data with the results obtained through Finite Element Analysis. A case study is also presented for each class of flexures, concerning the potential applications in the optimal design of planar and spatial compliant mechanisms. Each case study is followed by comparing the performance of these novel flexures with the performance of commonly used geometries in terms of principle compliance factors, parasitic motions and maximum stress demands. Furthermore, we have extended our study to the design and analysis of serial and parallel compliant mechanisms, where the proposed flexures have been employed to achieve spatial motions e.g. compliant spherical joints.

Analisi di interruzioni nelle metallizzazioni di celle fotovoltaiche attraverso simulazioni circuitali a modello distribuito

Studio delle variazioni delle prestazioni della cella solare a causa di non omogeneità localizzate nella struttura della cella (in questo caso trattasi di interruzioni nei fingers) . Valutazioni fatte per mezzo di simulazioni a rete elettrica distribuita, utilizzando un modello elettrico sviluppato in precedenza presso il laboratorio di Ingegneria di Cesena.

Danger assessment of a spring-gun device (muzzleloader) loaded only with black powder

We report a case involving a spring-gun device (muzzleloader) loaded solely with gunpowder, installed next to shoes to prevent the neighbors' puppy from removing them. The booby trap was triggered by the 15-year-old dog-owners son when he tried to put the shoes out of the reach of the puppy. The boy suffered second degree superficial burns located mainly at the dorsal side of the right hand and fingers. To estimate the danger of the used weapon, several tests were undertaken on soap blocks from different distances and with different loads of black powder. The particle density per mm and the depth of black powder tattooing in the soap was compared with the boy's injuries, and found conclusive with the gun-owners statement regarding the loading of the weapon. Furthermore, our experiments indicated that the gunpowder load involved was not able to inflict permanent damage, not even to the eyes, at the here estimated firing distance.

Fingering instability down the outside of a vertical cylinder

We present an experimental and numerical study examining the dynamics of a gravity-driven contact line of a thin viscous film traveling down the outside of a vertical cylinder of radius R. Experiments on cylinders with radii ranging between 0.159 and 3.81 cm show that the contact line is unstable to a fingering pattern for two fluids with differing viscosities, surface tensions, and wetting properties. The dynamics of the contact line is studied and results are compared to previous studies of inclined plane experiments in order to understand the influence substrate curvature plays on the fingering pattern. A lubrication model is derived for the film height in the limit that ε = H/R≪1, where H is the upstream film thickness, and in terms of a Bond number ρgR3/(γH), and the linear stability of the contact line is analyzed using traveling wave solutions. Curvature controls the capillary ridge height of the traveling wave and the range of unstable wavelength when ε = O(10-1), whereas the shape and stability of the contact line converge to the behavior one observes on a vertical plane when ε ≤ O(10-2). The most unstable wave mode, cutoff wave mode for neutral stability, and maximum growth rate scale as 0.45 where = ρgR2/γ ≥ 1.3, and the contact line is unstable to fingering when ≥ 0.56. Using the experimental data to extrapolate outside the range of validity of the thin film model, we estimate the contact line is stable when <0.56. Agreement is excellent between the model and the experimental data for the wave number (i.e., number of fingers) and wavelength of the fingering pattern that forms along the contact line.