The lectin-like domain of TNF protects from listeriolysin-induced hyperpermeability in human pulmonary microvascular endothelial cells — A crucial role for protein kinase C-α inhibition

Listeriosis can lead to potentially lethal pulmonary complications in newborns and immune compromised patients, characterized by extensive permeability edema. Listeriolysin (LLO), the main virulence factor of Listeria monocytogenes, induces a dose-dependent hyperpermeability in monolayers of human lung microvascular endothelial cells in vitro. The permeability increasing activity of LLO, which is accompanied by an increased reactive oxygen species (ROS) generation, RhoA activation and myosin light chain (MLC) phosphorylation, can be completely inhibited by the protein kinase C (PKC) α/β inhibitor GÖ6976, indicating a crucial role for PKC in the induction of barrier dysfunction. The TNF-derived TIP peptide, which mimics the lectin-like domain of the cytokine, blunts LLO-induced hyperpermeability in vitro, upon inhibiting LLO-induced protein kinase C-α activation, ROS generation and MLC phosphorylation and upon restoring the RhoA/Rac 1 balance. These results indicate that the lectin-like domain of TNF has a potential therapeutic value in protecting from LLO-induced pulmonary endothelial hyperpermeability.     

A systematic interaction map of validated kinase inhibitors with Ser/Thr kinases

Protein kinases play a pivotal role in cell signaling, and dysregulation of many kinases has been linked to disease development. A large number of kinase inhibitors are therefore currently under investigation in clinical trials, and so far seven inhibitors have been approved as anti-cancer drugs. In addition, kinase inhibitors are widely used as specific probes to study cell signaling, but systematic studies describing selectivity of these reagents across a panel of diverse kinases are largely lacking. Here we evaluated the specificity of 156 validated kinase inhibitors, including inhibitors used in clinical trials, against 60 human Ser/Thr kinases using a thermal stability shift assay. Our analysis revealed many unexpected cross-reactivities for inhibitors thought to be specific for certain targets. We also found that certain combinations of active-site residues in the ATP-binding site correlated with the detected ligand promiscuity and that some kinases are highly sensitive to inhibition using diverse chemotypes, suggesting them as preferred intervention points. Our results uncovered also inhibitor cross-reactivities that may lead to alternate clinical applications. For example, LY333'531, a PKCbeta inhibitor currently in phase III clinical trials, efficiently inhibited PIM1 kinase in our screen, a suggested target for treatment of leukemia. We determined the binding mode of this inhibitor by x-ray crystallography and in addition showed that LY333'531 induced cell death and significantly suppressed growth of leukemic cells from acute myeloid leukemia patients.     

Severe cardiomyopathy following treatment with the tumour necrosis factor‐α inhibitor adalimumab for Crohn's disease

Adalimumab belongs to the group of tumour necrosis factor‐α inhibitors and has been approved for the treatment Crohn's Disease since 2007. Herein we report a severe adverse reaction to adalimumab in a 25‐year‐old female patient. One week after the initial‐dose of adalimumab (160 mg), which was initiated due to an acute exacerbation of Crohn's disease, the patient developed a fulminant cardiomyopathy. In severe cardiogenic shock, the patient required an extracorporeal membrane‐oxygenation system for 8 days until cardiac recovery.     

Sympathetic skin response: monitoring of CT-guided lumbar sympathetic blocks

PURPOSE: To prospectively evaluate accuracy of sympathetic skin response (SSR) for monitoring computed tomography (CT)-guided lumbar sympathetic blocks, with palpable temperature increase in the foot 30 minutes after injection serving as the reference standard. MATERIALS AND METHODS: Institutional review board approval and written informed consent were obtained. Seventy individual lumbar sympathetic blocks were performed in 13 patients (six female, seven male; mean age, 45 years) with reflex sympathetic dystrophy of the foot. A 22-gauge needle was advanced to the sympathetic trunk at midlumbar level with CT fluoroscopic guidance, and 1 mL of iopamidol (200 mg of iodine per milliliter) and 5 mL of 0.5% bupivacaine were injected. SSR was monitored in both feet before and after bupivacaine injection. SSRs were activated with painless low-strength (5-20-mA) electrical stimuli. SSR ratio (SSR in the injected foot versus SSR in the contralateral foot) was calculated before injection and repeatedly at 1-minute intervals thereafter. Needle tip position and distribution of bupivacaine were measured on CT images. Receiver operating characteristic curves for SSR ratio were calculated until 7 minutes after injection. Logistic regression analyses adjusted for clustering were calculated for SSR ratio, injection parameters, needle tip position, and bupivacaine distribution. RESULTS: Thirty minutes after injection, 83% of procedures were considered clinically successful. An SSR cutoff ratio of 1:10 was used, and sensitivity, specificity, and accuracy of SSR for prediction of clinical success were 84%, 92%, and 86%, respectively, 4 minutes after injection and 95%, 92%, and 94%, respectively, 7 minutes after injection. Needle tip position (P = .19), medial and lateral borders of bupivacaine distribution (P = .11 and .056), and distance between bupivacaine distribution and the vertebral body (P = .41) were not significantly different between successful and unsuccessful injections. CONCLUSION: SSR can be used to correctly identify needle tip position in lumbar sympathetic blocks 6 and 7 minutes after injection.     

Inflammatory tumour of the prostate in a 4-year-old boy

An inflammatory tumour of the prostate is very rare in young boys without genital anomalies. We report a 4.5-year-old boy presenting with pollakisuria, dysuria, secondary urinary incontinence and decreased urine stream. Ultrasonography, magnetic resonance imaging and transrectal tumour biopsy justified the diagnosis of an inflammatory tumour of the prostate with a size of 5 × 3 × 5 cm. Under antibiotic therapy the clinical symptoms disappeared within 6 weeks. Three months later ultrasonography revealed a normal prostate with a residual midline cyst of 3 × 4 mm. Follow-up examinations for 2 years did not show any evidence of relapse. We believe that the inflammatory tumour can be explained by the embryologic development of the prostate and the persistence of an intraprostatic cyst.