Identification of hepta-histidine as a candidate drug for Huntington's disease by in silico-in vitro- in vivo-integrated screens of chemical libraries.

We identified drug seeds for treating Huntington's disease (HD) by combining in vitro single molecule fluorescence spectroscopy, in silico molecular docking simulations, and in vivo fly and mouse HD models to screen for inhibitors of abnormal interactions between mutant Htt and physiological Ku70, an essential DNA damage repair protein in neurons whose function is known to be impaired by mutant Htt. From 19,468 and 3,010,321 chemicals in actual and virtual libraries, fifty-six chemicals were selected from combined in vitro-in silico screens; six of these were further confirmed to have an in vivo effect on lifespan in a fly HD model, and two chemicals exerted an in vivo effect on the lifespan, body weight and motor function in a mouse HD model. Two oligopeptides, hepta-histidine (7H) and Angiotensin III, rescued the morphological abnormalities of primary neurons differentiated from iPS cells of human HD patients. For these selected drug seeds, we proposed a possible common structure. Unexpectedly, the selected chemicals enhanced rather than inhibited Htt aggregation, as indicated by dynamic light scattering analysis. Taken together, these integrated screens revealed a new pathway for the molecular targeted therapy of HD

Mechanisms underlying clinical efficacy of Angiotensin II type 2 receptor (AT(2)R) antagonist EMA401 in neuropathic pain: clinical tissue and in vitro studies

BACKGROUND: The clinical efficacy of the Angiotensin II (AngII) receptor AT(2)R antagonist EMA401, a novel peripherally-restricted analgesic, was reported recently in post-herpetic neuralgia. While previous studies have shown that AT(2)R is expressed by nociceptors in human DRG (hDRG), and that EMA401 inhibits capsaicin responses in cultured hDRG neurons, the expression and levels of its endogenous ligands AngII and AngIII in clinical neuropathic pain tissues, and their signalling pathways, require investigation. We have immunostained AngII, AT(2)R and the capsaicin receptor TRPV1 in control post-mortem and avulsion injured hDRG, control and injured human nerves, and in cultured hDRG neurons. AngII, AngIII, and Ang-(1-7) levels were quantified by ELISA. The in vitro effects of AngII, AT(2)R agonist C21, and Nerve growth factor (NGF) were measured on neurite lengths; AngII, NGF and EMA401 effects on expression of p38 and p42/44 MAPK were measured using quantitative immunofluorescence, and on capsaicin responses using calcium imaging. RESULTS: AngII immunostaining was observed in approximately 75% of small/medium diameter neurons in control (n = 5) and avulsion injured (n = 8) hDRG, but not large neurons i.e. similar to TRPV1. AngII was co-localised with AT(2)R and TRPV1 in hDRG and in vitro. AngII staining by image analysis showed no significant difference between control (n = 12) and injured (n = 13) human nerves. AngII levels by ELISA were also similar in control human nerves (4.09 ± 0.36 pmol/g, n = 31), injured nerves (3.99 ± 0.79 pmol/g, n = 7), and painful neuromas (3.43 ± 0.73 pmol/g, n = 12); AngIII and Ang-(1-7) levels were undetectable (<0.03 and 0.05 pmol/g respectively). Neurite lengths were significantly increased in the presence of NGF, AngII and C21 in cultured DRG neurons. AngII and, as expected, NGF significantly increased signal intensity of p38 and p42/44 MAPK, which was reversed by EMA401. AngII mediated sensitization of capsaicin responses was not observed in the presence of MAP kinase inhibitor PD98059, and the kinase inhibitor staurosporine. CONCLUSION: The major AT(2)R ligand in human peripheral nerves is AngII, and its levels are maintained in injured nerves. EMA401 may act on paracrine/autocrine mechanisms at peripheral nerve terminals, or intracrine mechanisms, to reduce neuropathic pain signalling in AngII/NGF/TRPV1-convergent pathways

On-line electrogeneration of copper-peptide complexes in microspray mass spectrometry

The interaction of copper ions with peptides was investigated by electrospray mass spectrometry. Two electrospray micro-emitters were compared, the first one with a platinum electrode using a copper(II) electrolyte solution containing a peptide sample, and the second one with a sacrificial copper anode in a water/methanol solution containing only a peptide (i.e., angiotensin III, bradykinin, or Leu-enkephalin). The former yielded mainly Cu2+ complexes either with histidine residues or with the peptide backbone (Cu+ complexes can be also formed due to gas-phase reactions), whereas the latter can generate a mixture of both Cu+ and Cu2+ aqueous complexes that yield different complexation patterns. This study shows that electrospray emitters with soluble copper anodes enable the study of Cu(I)-peptide complexes in solutio

Development of an animal-borne blood sample collection device and its deployment for the determination of cardiovascular and stress hormones in phocid seals

The research was supported by Bilateral Program between Japan and the United Kingdom and a Grant-in-Aid for Scientific Research (23247010) to Y. Takei and by Grant-in-Aid for challenging Exploratory Research (15K14567) to I. Suzuki from the Japan Society for the Promotion of Science. This work was also supported by funding from the U.K. Natural Environment Research Council (Grant SMRU1001).An animal-borne blood sampler with data-logging functions was developed for phocid seals, which collected two blood samples for the comparison of endocrino-logical/biochemical parameters under two different conditions. The sampler can be triggered by preset hydrostatic pressure, acceleration (descending or ascending), temperature, and time, and also man-ually by light. The sampling was reliable with 39/50 (78%) successful attempts to collect blood samples. Contamination of fluids in the tubing to the next blood sample was 1%, following the prior clearance of the tubing to a waste syringe. In captive harbor seals (Phoca vitulina), the automated blood-sampling method was less stressful than direct blood withdrawal, as evidenced by lower levels of stress hormones (P < 0.05 for ACTH and P = 0.078 for cortisol). HPLC analyses showed that both cortisol and cortisone were circu-lating in seal blood. Using the sampler, plasma levels of cardiovascular hormones, atrial natriuretic peptide (ANP), AVP, and ANG II were compared in grey seals (Halichoerus grypus), between samples collected when the animals were on land and in the water. HPLC analyses determined that [Met12] ANP (1-28) and various forms of angiotensins (ANG II, III, and IV) were circulating in seal blood. Although water immersion profoundly changes the plasma levels of cardiovascular hormones in terrestrial mammals, there were only tendencies toward an increase in ANP (P = 0.069) and a decrease in AVP (P = 0.074) in the seals. These results suggest that cardiovascular regulation in phocid seals may have undergone adaptation during evolution of the carnivore to a semiaquatic lifestyle.PostprintPeer reviewe

Welsh and immigrant new firms and new firm founders : are they really different?

School of Managemen

Flight data display studies for real time computer flight evaluation Final report

Real time displays for in-flight monitoring of Saturn launch vehicle

Uniwersytet - zarys ewolucji idei podstawowej

Two basic ideas of what a university should be have competed throughout history. The oldest conception of the university can be traced to Plato’s Academy. This idea was later developed by Wilhelm von Humboldt and revived by Allan Bloom. According to this approach the university is a multi-functional institution which combines education with the pursuit of truth. Mediaeval universities, the Napoleonic university and the idea of university as a higher vocational school developed according to the assumptions of the newer model. There are also a number of intermediate approaches where the point of departure is the didactic function but the didactic process is based on the specific values of the older approach, i.e., general and theoretic knowledge. These models were radically questioned in the second half of the twentieth century. The crisis of the university is partly a function of the unfavourable cultural climate (disbelief in the value of the scientific method and the validity of knowledge and learning) and also to some extent of the incoherent goals of university education (educating the intellectual versus educating the employee) and the faulty organisation of the university as an institution.Na przestrzeni dziejów ścierały się dwie podstawowe idee uniwersytetu. Najstarszą koncepcję uniwersytetu można dostrzec w Akademii Platońskiej, została ona później rozwinięta przez Wilhelma von Humboldta i przypomniana przez Allana Blooma. Uniwersytet jest tu postrzegany jako instytucja wielofunkcyjna, która łączy kształcenie z poszukiwaniem prawdy. Zgodnie z założeniami nowszej koncepcji powstały uniwersytety średniowieczne, uniwersytet napoleoński oraz idea uniwersytetu jako wyższej szkoły zawodowej. Istnieją również stanowiska pośrednie, dla których funkcja dydaktyczna stanowi punkt wyjścia, jednak sam proces dydaktyczny jest oparty na wartościach specyficznych, charakterystycznych dla starej koncepcji: na wiedzy ogólnej i teoretycznej. Idee uniwersytetu zostały w pełni zakwestionowane w drugiej połowie XX w. Kryzys uniwersytetu jest częściowo pochodną niesprzyjającego klimatu kulturowego: zwątpienia w wartość metody badań naukowych oraz w prawomocność wiedzy i nauki, a częściowo niekoherencji celów edukacji uniwersyteckiej: kształcenie inteligenta versus  kształcenie pracownika, oraz złej organizacji uniwersytetu jako instytucji

Pharmacological effects of raas blockade in ischemic nephropathy

Background: The management of ischemic nephropathy due to atherosclerotic renal artery stenosis has become increasingly conservative in the modern era, with current guidelines recommending optimized medical therapy as the initial step. The doubts raised by the recently published trials of revascularization strategies have led to a renewed focus on pharmacological strategies promoting blood pressure control and renal protection. It is essential to further elucidate the pathophysiological mechanisms underlying hypoperfusion induced renal microvascular dysfunction with subsequent tissue injury and fibrogenesis. The role of renin angiotensin aldosterone system as a mediator of the main pathophysiological consequences of ischemic nephropathy is well known. However, more recent experimental evidence on the adrenergic system and intrarenal tubular feedback mechanisms has stimulated new interest towards a multi-target therapeutic approach. Methods: This review focuses on the pharmacology of the principle therapeutic drug classes currently used in the treatment of atherosclerotic renal artery stenosis with an analysis of their metabolic aspects and use in clinical practice based on evidence from clinical trials. Results and Conclusions: An optimal pharmacologic approach is crucial for a successful prevention of renal injury and cardiovascular events in this high-risk population. Antihypertensive treatment should include renin angiotensin aldosterone system blockade medication not only for their antihypertensive properties, but especially for those cardio and renoprotectiv