Exploring the short linear motif-mediated protein-protein interactions of CrkL through ProP-PD.

Adaptor proteins play a pivotal role in cellular signaling mediating a multitude of protein-protein interaction critical for cellular homeostasis. Dysregulation of these interactions has been linked to the onset of various cancer pathologies and exploited by viral pathogens during host cell takeover. CrkL is an adaptor protein composed of an N-terminal SH2 domain followed by two SH3 domains that mediate interactions with diverse partners through the recognition of specific binding motifs. In this study, we employed proteomic peptide-phage display (ProP-PD) to comprehensively explore the short linear motif (SLiM)-based interactions of CrkL. Furthermore, we scrutinized how the binding affinity for selected peptides was influenced in the context of the full-length CrkL versus the isolated N-SH3 domain. Importantly, our results provided insights into SLiM-binding sites within previously reported interactors, as well as revealing novel human and viral ligands, expanding our understanding of the interactions mediated by CrkL and highlighting the significance of SLiM-based interactions in mediating adaptor protein function, with implications for cancer and viral pathologies.

Fatty acid profile, carcass and meat quality traits of young Nellore bulls fed crude glycerin replacing energy sources in the concentrate.

Carcass and meat quality traits of 60 Nellore young bulls fed diets without crude glycerin (CG); with CG replacing corn (CGc; 10% of dry matter - DM) in the concentrate; and with CG replacing soybean hull (CGsh; 10% of DM) in the concentrate were evaluated. Diets were evaluated at two concentrate levels (CLs). The CL did not affect cold carcass weight (CCW; P=0.6074), cold carcass dressing (CCD; P=0.9636), rib fat thickness (RFT; P=0.8696) and longissimus muscle area (LMA; P=0.7524). Animals fed diets with CGc or CGsh showed meat with greater deposition of monounsaturated fatty acid (MUFA; P=0.0022) and CLA (18:2 cis-9, trans-11) contents (P=0.0001) than animals fed diets without CG. The inclusion of 10% of CG in diets CGc or CGsh does not affect the carcass and meat quality traits; however, it increases the MUFA and CLA contents in beef, although these changes are very small in nutritional terms.

In vitro and in vivo studies with tetra-hydro-jasmonic acid (LR2412) reveal its potential to correct signs of skin ageing.

BACKGROUND: LR2412, a synthetic derivative of jasmonic acid, improved the reconstruction and homeostasis of our organotypic skin models. OBJECTIVES: The need for efficient 'anti-ageing' treatments, in particular for the management of photoaged skin, prompted us to investigate this new ingredient for its potential to correct signs of skin ageing in vitro and in vivo and to identify its mode of action. RESULTS: In vitro, penetration of LR2412 was evaluated using a Franz diffusion cell on excised human skin. Its exfoliating properties and interactions with the stratum corneum were studied using electron microscopy and X-ray diffraction. Experiments were performed on a human reconstructed skin model. In vivo, the effects of LR2412 on steroid-induced skin atrophy, a clinical skin ageing model, were assessed vs. vehicle. A patch test study evaluated its effect on deposition of fibrillin-rich microfibrils in the papillary dermis in clinically photoaged volunteers. A clinical study on the appearance of crow's feet wrinkles was conducted over 3 months of daily application. Penetration studies revealed that LR2412 reaches viable epidermis and superficial dermis, which are skin targets of anti-ageing actives. Within the upper layers of the stratum corneum LR2412 accelerates desquamation and improves the mechanical properties. At the dermal-epidermal junction of reconstructed skin, collagen IV, laminin-5 and fibrillin were stimulated. In vivo, LR2412 reversed steroid-induced atrophy. The patch test model confirms the deposition of fibrillin-rich microfibrils, then an in use clinical study revealed that it reduced facial wrinkles. CONCLUSIONS: The in vitro and in vivo data demonstrate that based on its multiple interactions within human skin, LR2412 has potential to partially correct the signs of ageing in intrinsically and photoaged skin.

Management of unruptured brain aneurysms: retrospective analysis of a single centre experience.

Embolization is very effective in preventing bleeding of unruptured aneurysms with lower rates of mortality and morbidity than surgical treatment. Neurosurgery remains, however, a good alternative. This retrospective analysis examined data stored the digital database of Bellaria Hospital Radiology Department, evaluating patients, treatments and devices used as well as outcomes and complications. Therapy should be tailored to each individual case to offer each patient the best treatment. Out of 265 unruptured intracranial aneurysms detected, 182 were treated by embolization. 16 cases presented complications (12 only radiologically found); severe clinical consequences occurred in 3%: one ischaemia and five haemorrhages. Conservative treatment was adopted in 21 patients. Long-term follow-up is mandatory as aneurysms could increase their size and develop an irregular morphology in ten years' time. Endovascular embolization is a very effective treatment with positive outcomes in the majority of cases and a percentage of retreatments of 8%. In order to increase the number of successful cases, a multidisciplinary cooperation with neurosurgeons is strongly recommended.

CT angiography in an acute stroke protocol: correlation between occlusion site and outcome of intravenous thrombolysis.

Thrombolysis with intravenous rt-PA is the current therapy for acute ischemic stroke. Unlike other outcome factors, relatively little is known about the prognostic value of the occlusion site on treatment outcome. We compared the effectiveness and safety of intravenous thrombolysis in patients with different levels of occlusion identified by CT angiography (CTA) in anterior circulation stroke, and analyzed the influence of the occlusion site on treatment outcome in relation to other outcome factors. We selected 71 patients from a stroke database collected between June 2007 and December 2011 at our hospital. All of the studied patients had anterior circulation stroke with intracranial occlusion detected by CTA and were treated with intravenous rt-PA. They were divided into two groups according to the site of occlusion along the middle cerebral artery course: proximal (carotid "T", complete M1 and mild M1 occlusions) and distal (M2/M3 occlusions). Treatment effectiveness was assessed by modified Rankin Scale (mRS) at three months, considering a positive outcome a mRS value ≤ 2. Treatment safety was assessed by evaluating the rate of hemorrhagic complications seen on unenhanced CT at 24 hours. Binary logistic regression was performed to evaluate the interaction between occlusion site and other variables such as sex, age, ASPECT score on admission and baseline NIHSS value in determining treatment outcome. The degree of effectiveness and safety differed when considering patients with proximal and distal occlusions. The percentage of successfully treated cases was 28.6% in the first group compared to 72% in the second, and the rate of hemorrhagic complications was 28.6% and 6% respectively. After adjustment for sex, age, ASPECT score on admission and baseline NIHSS value, occlusion site was the only variable significantly influencing treatment safety and, together with baseline NIHSS value, the only valid predictor of treatment effectiveness. We demonstrated a correlation between the site of arterial occlusion and outcome of intravenous thrombolysis. By helping the choice of the best therapeutic strategy depending on the identified occlusion site, CTA could be usefully added to the examinations included in the Stroke Protocol for the baseline evaluation of patients with suspected acute stroke.

Aneurysmal bone cyst of the mobile spine: the therapeutic role of embolization.

PURPOSE: Our aim is to define the role of embolization in the treatment of aneurysmal bone cyst of the spine in order to include this option in the decision making process. METHODS: From April 2004 to November 2009, seven patients with primary aneurysmal bone cyst of the mobile spine treated by embolization have been prospectively followed-up. All clinical presentations and imagings were recorded. There are many options of embolic agent and techniques used, but all aim to devascularize the tumor. The therapeutic protocol includes: embolization repeated every 8 weeks until the appearance of radiographic signs of healing. Complications, rate of healing and clinical outcome were analyzed. RESULTS: The number of embolizations varied from one to a maximum of seven without related intra- or post-operative complications. One patient, after four selective arterial embolizations, underwent direct percutaneous injection of embolic agents into the cyst. A clinical and radiographical response was achieved in all patients who were found alive and completely free of disease at mean follow-up of 46 months after last treatment and nobody crossed to surgical option. CONCLUSION: Embolization seems to be the first option for spinal aneurysmal bone cyst treatment because of the best cost-to-benefit ratio. It is indicated in intact aneurysmal bone cyst, when diagnosis is certain, when technically feasible and safe and when no pathologic fracture or neurologic involvements are found. If embolization fails, other options for treatment would still be available.

Complications in the treatment of intracranial aneurysms with silk stents: an analysis of 30 consecutive patients.

Flow-diverting stents (Silk and PED) have radically changed the approach to intracranial aneurysm treatment from the use of endosaccular materials to use of an extraaneurysmal endoluminal device. However, much debate surrounds the most appropriate indications for the use of FD stents and the problems raised by several possible complications.We analysed our technical difficulties and the early (less than ten days after treatment) and late complications encountered in 30 aneurysms treated comprising 13 giant lesions, 12 large, five with maximum diameters <10 mm and one blister-like aneurysm. In our experience the primary indications for the use of FD stents can be the symptomatic intracavernous giant aneurysms. Although the extracavernous carotid siphon aneurysms have major risk of bleeding, FD stents are indicated clearly explaining the risks to the patient in case of severe mass effect. There is a very complex assessment for aneurysms of the vertebrobasilar circulation.

Manejo quirúrgico del síndrome de silla turca vacía primaria con compromiso visual campimétrico y sin evidencia de herniación del sistema visual / Surgical treatment of primary empty sella syndrome with visual compromise and no evidence of visual system herniation

Presentamos el caso de una mujer de 36 años con síndrome de silla turca vacía primaria (STVP) caracterizado por cefalea, estrechamiento concéntrico periférico progresivo de la visión y oligomenorrea, quien fue sometida a remodelamiento selar con colocación de un autoinjerto intraselar. La evolución postoperatoria fue con mejoría importante del defecto campimétrico, en ambos ojos.

We report the case of a 36 year old woman with primary empty sella syndrome (PESS) and symptoms consisting in headache, progressive concentric peripheral narrowing of vision and oligomenorrhea, who underwent sellar remodeling with placement of an intrasellar autograft. Post operative course showed bilateral improvement in campimetric defect.

A minimally invasive treatment for lumbar disc herniation: DiscoGel® chemonucleolysis in patients unresponsive to chemonucleolysis with oxygen-ozone.

A multitude of therapies is available to treat disc herniation, ranging from conservative methods (medication and physical therapy) to minimally invasive (percutaneous) treatments and surgery. O2-O3 chemonucleolysis (O2-O3 therapy) is one of the minimally invasive treatments with the best cost/benefit ratio and lowest complication rate. Another substance recently made available exploiting the chemical properties of pure ethanol is DiscoGel®, a radiopaque gelified ethanol more viscous than absolute alcohol 8,9. The present study aimed to assess the therapeutic outcome of DiscoGel® chemonucleolysis in patients with lumbar disc herniation unresponsive to O2-O3 therapy. Thirty-two patients aged between 20 and 79 years were treated by DiscoGel® chemonucleolysis between December 2008 and January 2010. The treatment was successful (improvement in pain) in 24 out of 32 patients. DiscoGel® is safe and easy to handle and there were no complications related to product diffusivity outside the treatment site. The therapeutic success rate of DiscoGel® chemonucleolysis in patients unresponsive to O2-O3 therapy was satisfactory. Among other methods used to treat lumbar disc herniation, DiscoGel® chemonucleolysis can be deemed an intermediate procedure bridging conservative medical treatments and surgery.

Treatment of intracranial aneurysms using flow-diverting silk stents (BALT): a single centre experience.

The Silk stent (Balt, Montmorency, France) is a retractable device designed to achieve curative reconstruction of the parent artery associated with an intracranial aneurysm. We present our initial experience with the Silk flow-diverting stent in the management and follow-up of 25 patients presenting with intracranial aneurysms.Twenty-five patients (age range, 34-81 years; 24 female) were treated with the Silk flow-diverting device. Aneurysms ranged in size from small (5), large (10) and giant (10) and included wide-necked aneurysms, multiple, nonsaccular, and recurrent intracranial aneurysms. Nine aneurysms were treated for headache, 14 for mass effect. None presented with haemorrhage. All patients were pretreated with dual antiplatelet medications for at least 72 hours before surgery and continued taking both agents for at least three months after treatment. A total of 25 Silk stents were used. Control MR angiography and/or CT angiography was typically performed prior to discharge and at one, three, six and 12 months post treatment. A follow-up digital subtraction angiogram was performed between six and 19 months post treatment.Complete angiographic occlusion or subtotal occlusion was achieved in 15 patients in a time frame from three days to 12 months. Three deaths and one major complication were encountered during the study period. Two patients, all with cavernous giant aneurysms, experienced transient exacerbations of preexisting cranial neuropathies and headache after the Silk treatment. Both were treated with corticosteroids, and symptoms resolved completely within a month.In our experience the Silk stent has proven to be a valuable tool in the endovascular treatment of intracranial giant partially thrombosed aneurysms and aneurysms of the internal carotid artery cavernous segment presenting with mass effect. The time of complete occlusion of the aneurysms and the risk of the bleeding is currently not predictable.

The use of flow-diverting stents in the treatment of giant cerebral aneurysms: preliminary results.

The treatment of giant cerebral aneurysms has always been a challenge for neurosurgeons and neuroradiologists. Flow-diverting stents (Silk; Pipeline Embolization Device) are new endovascular devices introduced for the treatment of intracranial aneurysms without release of intrasaccular coils. They are tubular bimetallic endoluminal devices with low porosity. We have employed these stents in the Neuroradiology Unit of Bellaria Hospital (Bologna, Italy) since the end of 2008, treating nine patients with giant carotid cerebral aneurysms using nine Silk stents as soon as the device obtained the CE mark. All patients were pretreated with dual antiplatelet medications before surgery. The Silk stents were deployed through a 4F Balt introducer, which ensured an uneventful and very quick procedure. Control CT angiography or MR angiography was typically performed at discharge and one, three, six and 12 months after treatment. Post-treatment results were: four complete occlusions, three near complete occlusions (residual neck flow) with reduced volume of the aneurysm and two more than 50% reduction of intra-aneurysmal flow. A fatal hemorrhagic complication occurred in one patient, probably due to the antiplatelet treatment. The Silk stent seems a very interesting curative device to treat giant aneurysms with preservation of the parent artery and small adjacent branches. Technical improvements will certainly reduce the thrombogenic effect with the related risks.

A Little Talk on Adamkiewicz's Artery. Some Practical Considerations on the Pre-Operative Identification of this Artery Starting from a Single Team Experience in Pre-Surgical Selective Embolization of Vascularized Spinal Lesions.

The major radicular artery eponymically named "Adamkiewicz's artery" (AKA) is an important vessel supplying the spinal cord, especially the lumbar enlargement. This report emphasizes the importance of anatomical knowledge of this artery and highlights the concept of the potential risk of neurological complications during different procedures: spine orthopedic/neurosurgery, aortic repair (vascular surgery) and endovascular selective embolizations performed by interventional neuro/radiologists. Anatomical considerations are made on the spinal cord arterial circulation with a special focus on the AKA. Our review of the literature considered this anatomical element essential to compare the potential risk of spinal cord ischemic damage during orthopedic/neurosurgical spine procedures, aortic vascular surgery repair procedures and endovascular selective arterial embolizations. Evaluation of the endovascular selective arterial spine embolization risk was based on our series of 410 embolization procedures. Spinal cord infarction and transient or permanent paraplegia may result from inadvertent interruption of the AKA. The presence of intersegmental collaterals may decrease the risk of spinal cord ischemia: this is an important element to bear in mind that may help in spine surgery or aortic repair procedures performed by vascular surgeons. Nevertheless, during aortic repair (open surgery or stent-graft procedures) interruption of bilateral segmental arteries at multiple consecutive levels including that of the AKA may occur thereby increasing the ischemic spinal cord risk, annulling the benefit of intersegmental collaterals. Accidental embolizations of the AKA during endovascular spine procedures (i.e. selective arterial embolizations) performed by interventional neuro/radiologists will cause an almost certain spinal cord infarction due to the consequent embolizations of the anterior spinal artery (ASA).

3T MRI in the Evaluation of Brain Aneurysms Treated with Flow-Diverting Stents: Preliminary Experience.

Deployment of stents across the neck of intracranial aneurysms to isolate the lesion from the circulation is a recently introduced endovascular treatment. These devices are known as flow-diverting stents because the stent mesh design drastically slows the blood flow within the aneurysm sac, thereby stimulating thrombus formation. Treated aneurysms require close follow-up monitoring using an effective minimally invasive method. We devised a dedicated follow-up protocol using a high field strength magnetic resonance system (MR) with gadolinium administration to monitor 11 patients treated by insertion of flow-diverting stents. Findings were compared with the results of a reference imaging procedure (CT angiography). MR accurately demonstrated patency of the stent lumen and monitored the evolution of the aneurysmal sac in all patients. Gadolinium administration proved essential in two patients to depict the complete exclusion of the flow within the aneurysmal sac.

Treatment of carotid siphon aneurysms with a microcell stent. A case report.

SUMMARY: The treatment of giant, large, multiple or wide-necked carotid siphon aneurysms has always represented a challenge for neurosurgeons and neuroradiologists. Very recently the use of stents with tiny holes has been proposed by two companies: Balt Silk Stent in Europe and Pipeline in America. We have used the Silk stent on a few patients and describe our first case who now has an eleven month follow-up. The carotid siphon presented three converging aneurysms sharing a very large common neck. The Silk stent (Balt Extrusion, Montmorency, France) was deployed through a 4F Balt introducer. The procedure was uneventful and very quick. As soon as the stent was positioned contrast medium stagnation was displayed within the aneurysm. The patient's post-operative course was normal and she was discharged three days later in good health.

Carotid Stenting without Angioplasty and without Protection: The Advantages of a Less Invasive Procedure.

SUMMARY: Endovascular stenting is a consolidated alternative to thrombendarterectomy in the treatment of extracranial carotid artery atheromasic stenosis. The most common complication of stenting is a distal embolism causing clinically silent orsymptomatic cerebral ischaemia. To prevent this complication distal embolism protection devices are often used but their effectiveness remains unsettled. In addition, there is some evidence that distal embolism may actually be triggered by the protection systems due to clot formationat their distal surface or in the intimal lesions these systems cause. Another rarer complicationis hyperperfusion syndrome arising during both stenting and thrombendarterectomy but more common in endovascular procedures. To avoid these complications the Neuroradiology Service at Bellaria Hospital (Bologna Local Health Trust) has devised a mini-invasive carotid stenting technique that does not require either distal embolism protection or angioplasty. The technique uses only the radial force exerted by the self-expanding stent to widen the atherosclerotic stenosis slowly and gradually. The goal of treatment has also changed from a prompt restoration of the atheromasic vessel's original calibre to slow transformation of the hemodynamic significance of the stenosis. The technique's success lies mainly in selecting the stenosis to treat using CT angiography to analyse plaque morphology and structure.We used the technique to treat 83 stenotic lesions in 75 patients. The study aims to describe and discussour experience.

Simplified superovulatory treatments in Corriedale ewes.

Two experiments were designed to evaluate the possibility of simplifying superovulatory treatments in Corriedale ewes with use of ovine FSH (oFSH). Ewes received intravaginal progestogen sponges for 14 days. In Experiment 1, several simplified schedules were tested. Ewes were treated with 176 NIH-FSH-S1 units' oFSH given as a single injection in saline, along with 500 IU eCG 48 h before sponge removal (Group A1), in four equal doses (B1), or given as a single injection in a polyvinylpyrrolidone vehicle (C1) 24 h before sponge removal. In Experiment 2, the simplified protocol that exhibited the most desirable results in Experiment 1 (A2) was compared with the same protocol, but using less oFSH (132 units) (B2) and with the most conventional protocol (176 units of oFSH in eight decreasing doses; C2). Estrus was detected and ewes were naturally mated. The ovarian response and embryo production were assessed on Day 6 after estrus. LH was measured at 6h intervals from pessary withdrawal. The onset of estrus and the pre-ovulatory LH surge were advanced (P<0.05) in ewes treated with FSH and eCG. In Experiment 1, protocol A1 produced a greater percentage of superovulated ewes compared to C1 (100.0 compared with 58.3%; P<0.05), increased ovulation rate (13.8 corpora lutea compared with 6.2 and 4.7 for B1 and C1, respectively; P<0.05), and tended to increase the number of transferable embryos compared to B1 (P=0.08). In Experiment 2, percentages of superovulated ewes and ovulation rates were similar among groups; however, Group A2 tended to have more large follicles (P=0.07) than C2. The number of transferable embryos was similar among the three treatments. In conclusion, the reduced-dose oFSH given once along with eCG is the most appropriate superovulatory treatment because it combines simplicity and a lesser dose of gonadotropin, which also implies a reduction in cost, without reducing embryo production.

Use of Glubran 2 acrylic glue in interventional neuroradiology.

INTRODUCTION: Glubran 2 is a cyanoacrylate-based synthetic glue modified by the addition of a monomer synthesized by the manufacturer. With this material it is possible to obtain the stability of endovascular embolization that is needed to treat tumours and vascular disease. MATERIAL AND METHODS: We report our 3-year experience of the use of Glubran for treating extracerebral tumours, spinal tumours, spinal arteriovenous malformations, and brain and spine dural fistulae. Glubran 2 was diluted with Lipiodol and injected in a continuous column with the flow rate monitored by seriography. The injection was stopped when retrograde flow was displayed in the afferent vessel. RESULTS: There were no periprocedural or subsequent clinical complications and the glue resulted in successful selective permanent occlusion with intralesional penetration similar to the angiographic features of microcatheterization. CONCLUSIONS: The embolization procedure was technically straightforward and relatively safe. However, Glubran 2 can be difficult to use and the procedure does carry major risks for patients. Glue injection requires in-depth study of the lesion, its circulation and the collateral circulation to avoid severe complications due to inappropriate use.

Intracranial stenting in the treatment of wide-necked aneurysms.

SUMMARY: We positioned the following self-expanding stents certified for intracranial application: 16 Neuroform (Boston Scientific), three INX (Medtronic), one Leo (Balt). 6F calibre femoral introducers and guiding catheters were used for stent placement changing to 5F calibre introducers and guiding catheters (Envoy, Cordis) for the Neuroform 2 and 3 stents. All procedures were carried out under general anaesthesia and heparinization. Our pharmacological protocol consisted of adjunctive treatment with anti-aggregants during the interventional procedure and for the following six months, without premedication. From November 2000 to August 2006 we treated 28 patients (27 F/1M) with giant wide-necked aneurysms and one dissecting basilar artery aneurysm requiring the placement of 29 stents. We successfully positioned 20 stents: 11 stents combined with coils (8 immediate; 3 late) with complete exclusion of the aneurysm from the circulation in seven cases and subtotal exclusion in four; nine stents not followed by embolization with complete exclusion of the aneurysm from the circulation in six cases and subtotal exclusion in three. Stenting was not possible in nine cases due to extreme vessel tortuosity and the poor flexibility of release systems for the first stents. No late stent occlusion or subarachnoid haemorrhage were encountered after treatment.

Brain AVM Embolization with Onyx(R): Analysis of Treatment in 34 Patients.

SUMMARY: The endovascular approach to arteriovenous malformations (AVM) using different embolizing agents is a well-established treatment option. This report assesses the results of our experience using a non "glueing" embolic material available for several years, commercially known as Onyx(R). We used Onyx to treat 34 consecutive patients in the last four years. All patients were treated in the same department by the same neuroradiological team, with a strictly repetitive technical strategy and procedural protocol. All our patients presented AVMs with Spetzler Grade 3 or more, because in our Institution Grade 1 or 2 AVMs are directly treated by surgical approach. We adopt a multidisciplinary treatment approach (embolization, surgery, radiotherapy) by which embolization is construed as work in progress offering definitive treatment of AVMs without severe risks. Embolization is mainly undertaken as the first step before surgery, to reduce flow and size of the AVM by a "targeted" technique. In addition to reducing lesion size, endovascular treatment aims to seal off AVM areas anatomically or haemodynamically complex for surgical treatment. Occasionally, the reduction in size allows a radiosurgical approach. Embolization seldom results in a definitive cure of AVMs. At the end of multimodal approach, we obtained the complete and definitive cure of AVM in 21/34 patients (two complete obliteration with interventional technique, 19 in combination with surgery); to these were added 5/34 patients who received radiosurgical therapy. No major complications arose during endovascular treatment. One patient had transitory (36 hour) impaired right arm pronation. The CT scan disclosed an asymptomatic mild SAH in the left sylvian fissure but no ischaemic areas. One patient still in treatment died from fatal rebleeding (the clinical onset had been with haemorrhage two weeks before the session) 12 days after the embolization. Excellent or good clinical outcome was obtained in 23/26 patients who completed the therapeutic protocol. Outcome was conditioned by focal symptoms present on admission in three patients due to haemorrhagic onset, but only one patient presented a severe disability on discharge. In our view, the main problem of Onyx is that the apparently easier approach will probably lead to a wider diffusion of these procedures. AVMs are extremely difficult and dangerous to treat: this is not affected by the quality of the embolizing agents used and must be kept in mind at all times.