Academic literature on the topic 'CR4056'

Recent evidence suggests that I2-imidazoline ligands have neuroprotective properties in animal models of neurodegeneration, such as Alzheimer’s disease (AD). We recently demonstrated that the I2-ligand BU224 reversed memory impairments in AD transgenic mice and this effect was not because of reductions in amyloid-β (Aβ) deposition. In this study, our aim was to determine the therapeutic potential of the powerful analgesic I2-imidazoline ligand CR4056 in the 5xFAD model of AD, since this ligand has been proven to be safely tolerated in humans. Sub-chronic oral administration of CR4056 (30 mg/kg for 10 days) led to an improvement in recognition memory in 6-NOur results also revealed a change in the profile of microglia by CR4056, resulting in a suppression of pro-inflammatory activated microglia, but increased the density of astrocytes and the expression of ApoE, which is mainly produced by these glial cells. In addition, CR4056 restored fibrinogen extravasation, affecting the distribution of markers of astrocytic end feet in blood vessels. Therefore, these results suggest that CR4056 protects against Aβ-mediated neuroinflammation and vascular damage, and offers therapeutic potential at any stage of AD.

L'osteoartrite (OA) è una patologia degenerativa cronica molto comune negli anziani. Il dolore rappresenta il sintomo di OA più invalidante e precoce per il paziente. Ad oggi non esistono terapie curative per questa patologia. Inoltre, i farmaci più comunemente usati per contrastare il dolore OA sono spesso associati ad effetti collaterali. CR4056, ligando dei recettori imidazolinici-2, è un nuovo farmaco analgesico dimostratosi efficace in diversi modelli animali di dolore. Gli scopi di questo progetto sono stati analizzare la progressione del dolore OA e valutare l'efficacia di CR4056, comparato ad un FANS standard (naproxene), in due modelli di OA in ratto, in grado di replicare le componenti dolorose e strutturali della patologia umana. L’OA è stata indotta mediante singola iniezione intra-articolare di 1 mg/50 μl di monosodio iodoacetato (MIA) o mediante rottura del menisco mediale (MMT) nel ginocchio destro di ratti maschi. L’iniezione di MIA causa la degenerazione della cartilagine, mediante inibizione locale della glicolisi, mentre la resezione del legamento collaterale mediale e del menisco mediale destabilizza l’articolazione, inducendo degradazione della cartilagine e alterazioni dell’osso subcondrale. Nei modelli MIA e MMT la soglia algogena meccanica è stata valutata come allodinia e rispettivamente iperalgesia primaria o secondaria. Sono stati poi determinati l’asimmetria statica o dinamica del peso supportato delle zampe posteriori ipsilaterale e contralaterale (HPWD), la funzionalità motoria e l’attività locomotoria. L’espressione di proteine correlate al dolore (GFAP, pp38, pERKs e Iba-1) è stata valutata nel midollo spinale lombare ipsilaterale e contralaterale o nei DRG L4 e L5 ipsilaterali. CR4056 (2, 6 e 20 mg/kg) e naproxene (10 mg/kg) sono stati somministrati come trattamenti acuti e subacuti in entrambi i modelli. L’iniezione di MIA ha indotto uno sviluppo significativo di iperalgesia meccanica primaria, allodinia meccanica e asimmetria statica e dinamica di HPWD; nessun cambiamento si è verificato nell’attività locomotoria. CR4056 (6 e 20 mg/kg) ha ridotto significativamente l’allodinia e l’iperalgesia, dopo trattamento acuto (7 e 14 giorni post MIA) e soprattutto subacuto (da 14 a 21 giorni post MIA), mentre il naproxene è stato efficace solo dopo trattamento subacuto. Entrambi i composti sono risultati privi di effetto sull’asimmetria statica e dinamica di HPWD. L’espressione di pp38 e pERKs nel midollo spinale lombare ipsilaterale non si è dimostrata differente tra sham e MIA. L’iniezione di MIA ha invece indotto un aumento significativo delle cellule microgliali attivate (Iba1+) nella corna dorsale ipsilaterale del midollo spinale L4. Il trattamento subacuto con entrambi i composti ha ridotto significativamente tale attivazione della microglia. L’operazione MMT ha indotto uno sviluppo significativo di una progressiva asimmetria statica di HPWD e di una stabile iperalgesia meccanica secondaria. Non sono stati invece evidenziati allodinia, asimmetria dinamica di HPWD e cambiamenti nell’attività locomotoria e funzionalità motoria. CR4056 (20 mg/kg) e naproxene hanno dimostrato un significativo effetto anti-iperalgesico solo dopo trattamento acuto (28 giorni post MMT). Invece, il trattamento subacuto (da 28 a 42 giorni post MMT) con CR4056 (6 mg/kg) ha ridotto significativamente l’asimmetria statica di HPWD, mentre il naproxene è risultato privo di effetti. Non si sono verificate variazioni nell’espressione di GFAP e Iba-1, rispettivamente nei DRG L4 e L5 ipsilaterali e nella corna dorsale ipsilaterale del midollo spinale L4, tra sham e MMT. Entrambi i modelli MIA e MMT sono caratterizzati da una componente di dolore OA comparabile a quella umana, con tuttavia un differente contributo dei meccanismi dolorosi periferici e centrali. Inoltre, i dati ottenuti dimostrano che CR4056 potrebbe costituire un nuovo efficace trattamento per il dolore OA.
Aim: Osteoarthritis (OA) is a disabling and painful condition very common in the elderly. Pain is the earliest symptom of OA. To date there are still no curative drugs for this condition. Moreover, the chronic use of first-line pharmacological treatments to handle OA pain is frequently associated with side effects. CR4056, an imidazoline-2 receptor ligand, is a promising analgesic drug that has been reported to be effective in several animal models of pain. The aims of my project were to analyze and compare the time-related progression of OA pain and to evaluate the efficacy of CR4056, in comparison with a standard NSAID (naproxen), in two well-established rat models of OA, able to mimic the painful and structural components of the human pathology. Methods: Knee OA was induced either by single intra-articular injection of 1 mg/50 μl monosodium iodoacetate (MIA) or by medial meniscal tear (MMT) in the right knee of male rats. The local injection of MIA produces cartilage degeneration, through the local inhibition of glycolisis, while the transection of both the medial collateral ligament and the medial meniscus leads to joint destabilization, resulting in cartilage degeneration and subchondral bone alterations. The withdrawal threshold to mechanical stimulation was assessed both as allodynia and either as primary or secondary hyperalgesia, in MIA and MMT model, respectively. Pain behaviour was further evaluated as static and dynamic hind paw weight bearing (HPWD) asymmetry between the ipsilateral and the contralateral limb, and as changes in motor function and/or locomotor activity. Pain-related proteins (GFAP, pp38, pERKs and Iba-1) expression was assessed in either ipsilateral and contralateral lumbar spinal cord or ipsilateral L4 and L5 dorsal root ganglions (DRGs), in either MIA or MMT model. CR4056 (2, 6 and 20 mg/kg) and 10 mg/kg naproxen were administered as acute and sub-acute treatments in both models. Results: MIA model was characterized by the significant development of primary mechanical hyperalgesia, mechanical allodynia and asymmetry in both static and dynamic HPWD. No changes were detected in locomotor activity after MIA injection. 6 and 20 mg/kg CR4056 significantly and dose-dependently reduced both allodynia and hyperalgesia, after acute (7 and 14 days after MIA) and especially after repeated treatment (from 14 to 21 days post-MIA), whereas naproxen was effective after sub-acute treatment only. Both compounds had no significant effect on static and dynamic HPWD changes. No difference was detected in pp38 and pERKs expression in ipsilateral lumbar spinal cord, between MIA and sham group. On the other hand, a significant increase in the number of Iba-1 positive, morphologically identified, activated microglia in ipsilateral L4 spinal cord dorsal horn occurred, 21 days after MIA injection. Sub-acute treatment with 6 mg/kg CR4056 and naproxen reversed MIA-induced microglia activation. MMT surgery induced the significant development of a progressive asymmetry in static HPWD and a long-lasting secondary mechanical hyperalgesia. No mechanical allodynia nor changes in dynamic HPWD, motor function and locomotor activity were detected after MMT surgery. 20 mg/kg CR4056 and naproxen promoted a mild but significant anti-hyperalgesic effect, after acute treatment (28 days post-surgery) only. Conversely, repeated treatment (from 28 to 42 days post-surgery) with 6 mg/kg CR4056 significantly reduced the progression of static HPWD asymmetry, whereas naproxen had no effects. No difference in GFAP or Iba-1 expression were detected, in either ipsilateral L4 and L5 DRGs or ipsilateral L4 spinal cord dorsal horn, between MMT and sham group. Conclusions: Both MIA and MMT OA models display a pain behaviour comparable to human OA, with however different relative contribution of peripheral and central pain mechanisms. Moreover, the data obtained showed that CR4056 may represent a new effective treatment option for OA pain.

Il dolore cronico è una patologia cronica invalidante attualmente trattata con farmaci prevalentemente oppioidi, che comportano gravi effetti collaterali come stitichezza, depressione respiratoria, dipendenza e tolleranza. I ligandi I2 possiedono proprietà analgesiche e contrastano lo sviluppo della tolleranza se co-somministrati con oppioidi. CR4056, un nuovo farmaco I2-agonista, possiede efficacia analgesica dimostrata in diversi modelli animali di dolore cronico, in cui, inoltre, è stato osservato un forte effetto sinergico con la morfina. Lo scopo del mio studio era quindi quello di esaminare l'interazione tra morfina e CR4056 per quanto riguarda la tolleranza agli oppioidi, per chiarire la loro interazione farmacologica. A questo scopo, ho valutato gli effetti comportamentali della sinergia tra CR4056 e morfina sullo sviluppo e l'espressione della tolleranza nel modello CFA di dolore cronico. E’ stata anche valutata l'azione putativa di CR4056 sugli effetti collaterali indotti da oppioidi e sulla microgliosi spinale. È noto, infatti, che la microglia spinale ha un ruolo importante nel dolore cronico e nella tolleranza indotta da oppioidi, poiché in queste condizioni è stata osservata un'attivazione prolungata della microglia, legata al rilascio di fattori proinfiammatori. Infine, abbiamo mirato a comprendere meglio la sinergia della morfina CR4056 a livello molecolare. Infatti, è stato dimostrato che i neuroni DRG di modelli animali di dolore cronico mostrano una maggiore attivazione del recettore TRPV1, dipendente della fosforilazione di PKCε e dalla sua traslocazione sulla membrana cellulare. Inoltre, molti analgesici sono in grado di inibire la fosforilazione di PKCε in neuroni sensoriali in coltura. In dati preliminari non pubblicati, abbiamo osservato che morfina e CR4056 contrastano la traslocazione di PKCε indotta da fattori infiammatori in colture primarie di DRG di ratto. Pertanto, in questo lavoro abbiamo mirato a chiarire gli effetti delle somministrazioni acute di morfina o CR4056 in DRG ex vivo da ratti trattati con CFA, mediante quantificazione della fosforilazione di PKCε e dell'espressione di TRPV1, e nel midollo spinale, mediante valutazione della microgliosi. I miei risultati hanno dimostrato che l'infiammazione indotta da CFA ha innescato l'iperalgesia meccanica, contrastata dalla morfina o dal CR4056 in acuto. La somministrazione combinata di morfina e CR4056 ha causato una prevenzione dose-dipendente della tolleranza alla morfina, rispetto al gruppo trattato con sola morfina, in un paradigma breve (4 giorni) e lungo (14 giorni). Inoltre, quando CR4056 è stato co-somministrato con la morfina in ratti già tolleranti, è stato in grado di migliorarne l'attività analgesica. Negli stessi animali, l'attivazione della microglia spinale era maggiore nei ratti CFA, trattati con veicolo o morfina, ma non nel gruppo con somministrazione combinata di CR4056-morfina. Inoltre, la sinergia non era accompagnata da una modulazione additiva della costipazione da oppioidi. Nei DRG L4-L5 di ratti CFA abbiamo riscontrato un aumento significativo della fosforilazione di PKC-ε e della colocalizzazione tra PKC-ε e VR1, rispetto ai controlli e agli animali trattati in acuto (con CR4056 o morfina). Negli stessi animali, la microglia spinale è risultata significativamente attivata negli animali trattati con CFA rispetto ai controlli e agli animali trattati con CR4056, ma non ai ratti trattati con morfina. In conclusione, questi dati suggeriscono CR4056 come farmaco valido per prevenire e revertire la tolleranza agli oppioidi senza esacerbarne gli effetti collaterali. Inoltre, i dati in vitro su CR4056 e il meccanismo sinergico con morfina su PKCε sono stati validati in DRG ex vivo da ratti trattati con CFA. Ulteriori studi chiariranno gli effetti del trattamento combinato su DRG e midollo spinale al fine di comprendere il meccanismo della sinergia tra CR4056 e oppioidi.
Chronic pain is a disabling and long-lasting cross-pathology condition, currently treated with mostly opioid drugs, which are leading to severe side-effects such as constipation, respiratory depression, addiction and tolerance. I2 ligands showed analgesic properties and to contrast tolerance development in co-administration with opioids. CR4056, a novel I2-agonist drug, has proved to have analgesic efficacy in several animal models of chronic pain, in which, moreover, a strong synergistic effect with morphine has been observed. The aim of my study was therefore to examine the interaction between morphine and CR4056 regarding opioid tolerance, and to elucidate their pharmacological interaction. To this scope, I assessed behavioral effects of the synergy between CR4056 and morphine on tolerance development and expression in the CFA model of chronic pain. Putative CR4056 action on opioid-induced side effects and spinal microgliosis were also assessed. It is known, in fact, that spinal microglia have an important role in chronic pain and opioid-induced tolerance, since, in these conditions, a sustained microglia activation has been observed, linked to the release of pro-inflammatory factors. Lastly, we aimed to better understand CR4056-morphine synergy at molecular level. In fact, it has been shown that DRG neurons of animal models of chronic pain display higher activation of TRPV1 receptor, depending on PKCε phosphorylation and translocation to cell membrane. Moreover, there is evidence that several analgesics are able to inhibit PKCε phosphorylation in cultured sensory neurons. Interestingly, in preliminary unpublished data, we observed that morphine and CR4056 can contrast PKCε translocation induced by inflammatory factors in primary rat DRG cultures. Therefore, in this work we aimed to elucidate the effects of acute administrations of morphine or CR4056 in ex vivo DRG from CFA-treated rats, by quantification of PKCε phosphorylation and TRPV1 expression, and in the spinal cord, by evaluation of microgliosis. My results demonstrated that CFA-induced inflammation triggered mechanical hyperalgesia, acutely counteracted by morphine or CR4056. Combined administration of morphine with CR4056 caused a dose-dependent prevention of morphine tolerance, which was established in the morphine alone treated group, in a short (4 days) and in a long paradigm (14 days). Moreover, when CR4056 was co-administered with morphine in already tolerant rats, it was able to improve morphine analgesic activity. In the same animals, spinal microglia activation was augmented in CFA-injected rats, either vehicle- or morphine-treated, but not in the group with CR4056-morphine combined administration. Besides, the synergy was not accompanied by an additive modulation of opioid-induced constipation. In L4-L5 DRG of CFA-injected rats we found a significant increase in the phosphorylation of PKC-ε, as well as in the colocalization between PKC-ε and VR1, compared to sham animals and to acutely treated animals (with CR4056 or morphine). In the same animals, activated microglial cells were significantly increased in CFA vehicle-treated animals compared to control and CR4056-treated animals, but not to morphine-treated rats. In conclusion, these data suggest that CR4056 seems to be a valid drug to prevent and rescue opioid tolerance without exacerbate side-effects. Moreover, in vitro data on CR4056 and morphine synergistic mechanism on PKCε were validated in ex vivo DRG from CFA-treated rats. Further studies will be needed to elucidate effects of combined treatment on DRG and spinal cord in order to understand the mechanism of CR4056 and opioids synergy.

This thesis seeks to illuminate the nature, extent and complexity of Templar interactions with their associates, particularly non-Christians, women and Mozarabs, by examining these interactions where the most evidence exists for them---northeastern Spain. Evidence for Temple associations with both Christians and non-Christians is strongest and most prolonged here. The overall nature of these interactions was friendlier than expected in a crusading group. In fact, Templars actively competed with the secular Church, nobility and the king in the Crown of Aragon for lordship over non-Christians because non-Christians were a lucrative tax base. Some non-Christians also sought association with the Templars because the Templars were a strong, international group with friendly ties to the Aragonese kings. The Temple could therefore offer protection from other lords against excessive taxation and exploitation, and physical attack. Documentary evidence shows mutually beneficial interactions as the Temple's (and its non-Christian associates') ongoing preference over time and space. Chapter one examines Templar interactions in general, both with associates and non-associates. Chapter two looks at Templar associations in Novillas, the first Templar house founded in the Crown of Aragon. Chapter three deals with the Tortosa and the lower Ebro Valley, which has the most varied surviving Templar documentation in the areas studied. Chapter four deals with Gardeny (in Lleida/Lerida), which has the largest number of surviving documents for all of the areas in the study. Chapter five looks at Monzon and Barcelona, the main Templar houses for Aragon and Catalonia respectively. The last chapter deals with Huesca, the northernmost house in the study.

This paper describes an ongoing effort to integrate Computational Fluid Dynamic methods into a system for predicting the performances of multi-component propulsors in a turbulent ship flow environment. These methods include a Reynolds-Averaged Navier-Stokes (RANS) code for ship turbulent flow calculations and a Vortex-Lattice Method (VLM) based program MPUF3A for sheet cavitating propeller flow analyses. The combination of these methods provides a new level of realism for modeling the flow field around a ship with thick turbulent boundary layers and multi-component propulsor/hull interaction. The bottlenecks caused by inadequate grid generation methods and a lack of code integration, which have prevented the use of these numerical methods for practical configurations in design time frame, have been substantially alleviated by the incorporation of zonal calculation techniques, advanced gridding tools, and a chimera gridding method that allows arbitrary grid block overlap. Validations of the calculation method were performed for a contra-rotating propeller, CR404 system, and a series 60, CB = 0.6, ship hull with a MAU propeller. Comparisons of the computational results to the measurement data are documented in this paper. Also, to demonstrate the predictive capability of the method, simulations were carried out for the series 60 ship hull with a single propeller and a contrarotating propeller under model and full scale conditions.