A mini-twister variant and impact of residues/cations on the phosphodiester cleavage of this ribozyme class.

Nucleolytic ribozymes catalyze site-specific cleavage of their phosphodiester backbones. A minimal version of the twister ribozyme is reported that lacks the phylogenetically conserved stem P1 while retaining wild-type activity. Atomic mutagenesis revealed that nitrogen atoms N1 and N3 of the adenine-6 at the cleavage site are indispensable for cleavage. By NMR spectroscopy, a pKa value of 5.1 was determined for a 13C2-labeled adenine at this position in the twister ribozyme, which is significantly shifted compared to the pKa of the same adenine in the substrate alone. This finding pinpoints at a potential role for adenine-6 in the catalytic mechanism besides the previously identified invariant guanine-48 and a Mg2+ ion, both of which are directly coordinated to the non-bridging oxygen atoms of the scissile phosphate; for the latter, additional evidence stems from the observation that Mn2+ or Cd2+ accelerated cleavage of phosphorothioate substrates. The relevance of this metal ion binding site is further emphasized by a new 2.6 Å X-ray structure of a 2′-OCH3-U5 modified twister ribozyme

'You can take a horse to water but you can't make it drink': Exploring children's engagement and resistance in family therapy

The final publication is available at Springer via http://dx.doi.org/10.1007/s10591-012-9220-8Children’s engagement and disengagement, adherence and non-adherence, compliance and non-compliance in healthcare have important implications for services. In family therapy mere attendance to the appointments is no guarantee of engaging in the treatment process and as children are not the main initiators of attendance engaging them through the process can be a complex activity for professionals. Through a conversation analysis of naturally occurring family therapy sessions we explore the main discursive strategies that children employ in this context to passively and actively disengage from the therapeutic process and investigate how the therapists manage and attend to this. We note that children competently remove themselves from therapy through passive resistance, active disengagement, and by expressing their autonomy. Analysis reveals that siblings of the constructed ‘problem’ child are given greater liberty in involvement. We conclude by demonstrating how therapists manage the delicate endeavour of including all family members in the process and how engagement and re-engagement are essential for meeting goals and discuss broader implications for healthcare and other settings where children may disengage

Case Reports1. A Late Presentation of Loeys-Dietz Syndrome: Beware of TGFβ Receptor Mutations in Benign Joint Hypermobility

Background: Thoracic aortic aneurysms (TAA) and dissections are not uncommon causes of sudden death in young adults. Loeys-Dietz syndrome (LDS) is a rare, recently described, autosomal dominant, connective tissue disease characterized by aggressive arterial aneurysms, resulting from mutations in the transforming growth factor beta (TGFβ) receptor genes TGFBR1 and TGFBR2. Mean age at death is 26.1 years, most often due to aortic dissection. We report an unusually late presentation of LDS, diagnosed following elective surgery in a female with a long history of joint hypermobility. Methods: A 51-year-old Caucasian lady complained of chest pain and headache following a dural leak from spinal anaesthesia for an elective ankle arthroscopy. CT scan and echocardiography demonstrated a dilated aortic root and significant aortic regurgitation. MRA demonstrated aortic tortuosity, an infrarenal aortic aneurysm and aneurysms in the left renal and right internal mammary arteries. She underwent aortic root repair and aortic valve replacement. She had a background of long-standing joint pains secondary to hypermobility, easy bruising, unusual fracture susceptibility and mild bronchiectasis. She had one healthy child age 32, after which she suffered a uterine prolapse. Examination revealed mild Marfanoid features. Uvula, skin and ophthalmological examination was normal. Results: Fibrillin-1 testing for Marfan syndrome (MFS) was negative. Detection of a c.1270G > C (p.Gly424Arg) TGFBR2 mutation confirmed the diagnosis of LDS. Losartan was started for vascular protection. Conclusions: LDS is a severe inherited vasculopathy that usually presents in childhood. It is characterized by aortic root dilatation and ascending aneurysms. There is a higher risk of aortic dissection compared with MFS. Clinical features overlap with MFS and Ehlers Danlos syndrome Type IV, but differentiating dysmorphogenic features include ocular hypertelorism, bifid uvula and cleft palate. Echocardiography and MRA or CT scanning from head to pelvis is recommended to establish the extent of vascular involvement. Management involves early surgical intervention, including early valve-sparing aortic root replacement, genetic counselling and close monitoring in pregnancy. Despite being caused by loss of function mutations in either TGFβ receptor, paradoxical activation of TGFβ signalling is seen, suggesting that TGFβ antagonism may confer disease modifying effects similar to those observed in MFS. TGFβ antagonism can be achieved with angiotensin antagonists, such as Losartan, which is able to delay aortic aneurysm development in preclinical models and in patients with MFS. Our case emphasizes the importance of timely recognition of vasculopathy syndromes in patients with hypermobility and the need for early surgical intervention. It also highlights their heterogeneity and the potential for late presentation. Disclosures: The authors have declared no conflicts of interes

Il mercato delle macchine agricole nel Sud-Est Europa (Bosnia - Erzegovina, Bulgaria, Croazia,Polonia, Romania, Serbia e Turchia)

Negli ultimi anni si è assistito ad un continuo spostamento sempre più ad est dei confini dell’Unione Europea, che si è dimostrata aperta ad accogliere i Paesi democratici europei in grado di affrontare le sfide economiche dell’adesione e di attuarne le norme. Dalla nascita ufficiale dell'UE, avvenuta nel 1993 e che coinvolgeva 12 Paesi, rapidamente si è passati a 15 (1995), per arrivare nel 2004 ad una UE a 25. A breve, l’1/1/2007, questi confini si allargheranno ulteriormente a seguito dell’accesso della Romania e della Bulgaria; si arriverà così ad un totale di 27 Paesi. Ma il fenomeno non si è ancora arrestato poiché altri Paesi ambiscono ad entrare nell’Unione,alcuni sono già candidati (Turchia e Croazia), per altri occorre ancora tempo e probabilmente, nel lungo periodo, i confini politici dell’UE coincideranno con i confini geografici dell’Europa. Nell’ambito di questo processo di allargamento ad est sono entrati o entreranno a far parte dell’Unione Paesi che fino a poco tempo fa non conoscevano l’economia di mercato e che nel giro di appena qualche decennio sono stati coinvolti nel processo di globalizzazione con l’apertura delle frontiere verso il mondo occidentale e l’avvento, anche nelle loro economie, del fenomeno del consumismo. In questo quadro appena delineato appare evidente che la struttura socio-economica e produttiva diquesti Paesi subirà dei grandi mutamenti, sicuramente seguendo la dinamica delle trasformazioniavvenute negli attuali Paesi industrializzati