Protease signaling through protease activated receptor 1 mediate nerve activation by mucosal supernatants from irritable bowel syndrome but not from ulcerative colitis patients.

BACKGROUND & AIMS: The causes of gastrointestinal complaints in irritable bowel syndrome (IBS) remain poorly understood. Altered nerve function has emerged as an important pathogenic factor as IBS mucosal biopsy supernatants consistently activate enteric and sensory neurons. We investigated the neurally active molecular components of such supernatants from patients with IBS and quiescent ulcerative colitis (UC). METHOD: Effects of supernatants from 7 healthy controls (HC), 20 IBS and 12 UC patients on human and guinea pig submucous neurons were studied with neuroimaging techniques. We identify differentially expressed proteins with proteome analysis. RESULTS: Nerve activation by IBS supernatants was prevented by the protease activated receptor 1 (PAR1) antagonist SCHE79797. UC supernatants also activated enteric neurons through protease dependent mechanisms but without PAR1 involvement. Proteome analysis of the supernatants identified 204 proteins, among them 17 proteases as differentially expressed between IBS, UC and HC. Of those the four proteases elastase 3a, chymotrypsin C, proteasome subunit type beta-2 and an unspecified isoform of complement C3 were significantly more abundant in IBS compared to HC and UC supernatants. Of eight proteases, which were upregulated in IBS, the combination of elastase 3a, cathepsin L and proteasome alpha subunit-4 showed the highest prediction accuracy of 98% to discriminate between IBS and HC groups. Elastase synergistically potentiated the effects of histamine and serotonin-the two other main neuroactive substances in the IBS supernatants. A serine protease inhibitor isolated from the probiotic Bifidobacterium longum NCC2705 (SERPINBL), known to inhibit elastase-like proteases, prevented nerve activation by IBS supernatants. CONCLUSION: Proteases in IBS and UC supernatants were responsible for nerve activation. Our data demonstrate that proteases, particularly those signalling through neuronal PAR1, are biomarker candidates for IBS, and protease profiling may be used to characterise IBS.

CT-guided brachytherapy as salvage therapy for intrahepatic recurrence of HCC after surgical resection.

AIM: Prevalence of patients with unresectable recurrence of hepatocellular carcinoma (HCC) after previous resection is rising. The purpose of this study was to determine survival of patients undergoing computed tomography-guided brachytherapy (CT-HDRBT). PATIENTS AND METHODS: Altogether 19 patients with unresectable HCC recurrence were treated with CT-HDRBT at our Institution. Patients underwent single-fraction high-dose irradiation by an iridium-192 source after CT fluoroscopy-guided catheter placement. The median tumor-enclosing target dose was 20 Gy. RESULTS: The median follow-up was 33 months. According to the Kaplan-Meier method, median overall survival after CT-HDRBT was 50 months, and median survival after first hepatic resection was 87 months. The median duration of local tumor control was 32 months and time to disease progression was 20 months. There were no serious complications after CT-HDRBT and no treatment-related deaths. CONCLUSION: CT-HDRBT is a safe, potentially life prolonging technique in patients with recurrence of HCC who have few therapeutic options.

Cerebrolysin protects PC12 cells from CoCl2-induced hypoxia employing GSK3ß signaling.

Cerebrolysin (EVER Neuro Pharma GmbH, Austria) is a peptidergic drug indicated for clinical use in stroke, traumatic brain injury and dementia. The therapeutic effect of Cerebrolysin is thought to ensure from its neurotrophic activity, which shares some properties with naturally occurring neurotrophic factors. However, the exact mechanism of action of Cerebrolysin is yet to be fully deciphered. This study aimed to investigate the neuroprotective effect of Cerebrolysin in a widely used in vitro model of hypoxia-induced neuronal cytotoxicity, namely cobalt chloride (CoCl2)-treatment of PC12 cells. CoCl2-cytotoxicity was indicated by a reduced cell-diameter, cell shrinkage, increased pro-apoptotic Caspase-activities and a decreased metabolic activity. Cerebrolysin maintained the cell-diameter of CoCl2-treated naïve PC12 cells, decreased the activation of Caspase 3/7 in CoCl2-stressed naïve PC12 cells and restored the cells' metabolic activity in CoCl2-impaired naïve and differentiated PC12 cells. Cerebrolysin treatment also decreased the levels of superoxide observed after exposure to CoCl2. Investigating the mechanism of action, we could demonstrate that Cerebrolysin application to CoCl2-stressed PC12 cells increased the phosphorylation of GSK3ß, resulting in the inhibition of GSK3ß. This might become clinically relevant for Alzheimer's disease, since GSK3ß activity has been linked to the production of amyloid beta. Taken together, Cerebrolysin was found to have neuroprotective effects in CoCl2-induced cytotoxicity in PC12 cells.

CT guided bone biopsy using a battery powered intraosseous device.

PURPOSE: To evaluate the feasibility of a battery powered intraosseous device to perform CT-fluoroscopy guided bone biopsy. METHODS: Retrospective analysis of 12 patients in whom bone specimen were acquired from different locations under CT-fluoroscopy guidance using the OnControl bone marrow biopsy system (OBM, Vidacare, Shavano Park, TX, USA). Data of the 12 were compared to a historic cohort in whom the specimen were acquired using the classic Jamshidi Needle, as reference needle using manual force for biopsy. RESULTS: Technical success was reached in 11 of 12 cases, indicated by central localisation of the needle within the target lesion. All specimen sampled were sufficient for histopathological workup. Compared to the historical cohort the time needed for biopsy decreased significantly from 13 ± 6 to 6 ± 4 min (P = 0.0001). Due to the shortened intervention time the radiation dose (CTDI) during CT-fluoroscopy was lowered significantly from 169 ± 87 to 111 ± 54 mGy × cm (P = 0.0001). Interventional radiologists were confident with the performance of the needle especially when using in sclerotic or osteoblastic lesions. CONCLUSION: The OBM is an attractive support for CT-fluoroscopy guided bone biopsy which is safe tool and compared to the classical approach using the Jamshidi needle leading to significantly reduced intervention time and radiation exposure.

Aging of the liver graft and functional quality in the absence of recurrent disease: a 10 year histological follow-up.

BACKGROUND: Although considerable information has been acquired concerning the function of older donor liver grafts it remains unclear how long these organs may fare well in their "cumulative life span". As graft challenge due to recurrent underlying disease can best be eliminated and controlled in patients transplanted for alcoholic liver disease (ALD) we analysed liver biopsies of ALD patients during a 10-year follow-up with special emphasis on donor age and parenchymal quality. MATERIAL/METHODS: Biopsies were taken from 271 patients 1, 3, 5, 10 years after LT. Specimen were analysed and staged concerning inflammation, rejection, fatty involution, and fibrosis/cirrhosis. Donor characteristics were recorded along with clinical and serological parameters, immunosuppressive protocols, rejection episodes, and patient and graft survival. RESULTS: 1, 3, 5 years after LT >80% of patients displayed only minimal fibrosis. In the further course fibrosis rates increased among all patients, with slightly more fibrosis for recurrent drinkers. Donor age did not influence fibrosis progression, long-term patient or graft survival. Regardless of alcohol abuse survival was excellent 5 years after LT, after 10 years abstinent patients did significantly better (82%;68%; p=0.017. CONCLUSIONS: LT for ALD offers excellent long-term survival with minimal histological alteration, especially in abstinent patients. Aged organs generate adequate function in ALD recipients.