Uptake of Clostridium botulinum C3 Exoenzyme into Intact HT22 and J774A.1 Cells

The Clostridium botulinum C3 exoenzyme selectively ADP-ribosylates low molecular weight GTP-binding proteins RhoA, B and C. This covalent modification inhibits Rho signaling activity, resulting in distinct actin cytoskeleton changes. Although C3 exoenzyme has no binding, the translocation domain assures that C3 enters cells and acts intracellularly. C3 uptake is thought to occur due to the high concentration of the C3 enzyme. However, recent work indicates that C3 is selectively endocytosed, suggesting a specific endocytotic pathway, which is not yet understood. In this study, we show that the C3 exoenzyme binds to cell surfaces and is internalized in a time-dependent manner. We show that the intermediate filament, vimentin, is involved in C3 uptake, as indicated by the inhibition of C3 internalization by acrylamide, a known vimentin disruption agent. Inhibition of C3 internalization was not observed by chemical inhibitors, like bafilomycin A, methyl-β-cyclodextrin, nocodazole or latrunculin B. Furthermore, the internalization of C3 exoenzyme was markedly inhibited in dynasore-treated HT22 cells. Our results indicate that C3 internalization depends on vimentin and does not depend strictly on both clathrin and caveolae.     

Video-based teleconsultations in pharmaceutical care – A systematic review

BackgroundIn recent years, telemedicine has gained increasing importance in the delivery of pharmaceutical care. The use of video technologies for remote communication between different parties offers the potential to meet the future challenges which arise from the increase in elderly and chronically ill patients. However, the influence of these technologies on patient-related outcomes in pharmaceutical care is not yet sufficiently known.MethodIn December 2018 a systematic literature search was conducted in the databases Medline, Cochrane Library and PubPharm. Randomized controlled trials were considered, which investigate real-time video conferencing between pharmacists on the one hand and patients or other healthcare providers on the other hand. The influence on patient related outcomes compared to standard care was assessed. The bias potential was evaluated using the Cochrane Risk-of-Bias instrument. A total of 4 randomized controlled trials could be included.ResultsThe studies describe partly complex intervention settings with adults and adolescents suffering from asthma, chronic renal failure, HIV infection, hyperlipidemia, hypertension and/or diabetes. None of the studies considers the interprofessional communication of pharmacists with other service providers. No influence on clinical or psychological endpoints was found. In some cases, an increase in adherence and correct medication use is evident. Healthcare utilization is not influenced. In all studies there is an increased risk of systematic bias.DiscussionTeleconsultations with pharmacists can rather be used to ensure the general provision of pharmaceutical care than to improve patient-related outcomes. Further studies are necessary to fully depict the influence of telemedical interventions in pharmaceutical care.     

A comparison of volume clamp method-based continuous noninvasive cardiac output (CNCO) measurement versus intermittent pulmonary artery thermodilution in postoperative cardiothoracic surgery patients

The CNAP technology (CNSystems Medizintechnik AG, Graz, Austria) allows continuous noninvasive arterial pressure waveform recording based on the volume clamp method and estimation of cardiac output (CO) by pulse contour analysis. We compared CNAP-derived CO measurements (CNCO) with intermittent invasive CO measurements (pulmonary artery catheter; PAC-CO) in postoperative cardiothoracic surgery patients. In 51 intensive care unit patients after cardiothoracic surgery, we measured PAC-CO (criterion standard) and CNCO at three different time points. We conducted two separate comparative analyses: (1) CNCO auto-calibrated to biometric patient data (CNCO_bio) versus PAC-CO and (2) CNCO calibrated to the first simultaneously measured PAC-CO value (CNCO_cal) versus PAC-CO. The agreement between the two methods was statistically assessed by Bland–Altman analysis and the percentage error. In a subgroup of patients, a passive leg raising maneuver was performed for clinical indications and we present the changes in PAC-CO and CNCO in four-quadrant plots (exclusion zone 0.5 L/min) in order to evaluate the trending ability of CNCO. The mean difference between CNCO_bio and PAC-CO was +0.5 L/min (standard deviation?±?1.3 L/min; 95% limits of agreement ?1.9 to +3.0 L/min). The percentage error was 49%. The concordance rate was 100%. For CNCOcal, the mean difference was ?0.3 L/min (±0.5 L/min; ?1.2 to +0.7 L/min) with a percentage error of 19%. In this clinical study in cardiothoracic surgery patients, CNCO_cal showed good agreement when compared with PAC-CO. For CNCO_bio, we observed a higher percentage error and good trending ability (concordance rate 100%).     

Cardiac Genetic Predisposition in Sudden Infant Death Syndrome

Background

Sudden infant death syndrome (SIDS) is a leading cause of postneonatal mortality. Genetic heart diseases (GHDs) underlie some cases of SIDS.

Modulation of platelet and leucocyte function by a Chinese herbal formulation as compared with conventional antiplatelet agents

Platelet and leucocyte activity are important in the acute development of thrombosis and in the pathogenesis of ischaemic vascular disease. Dan Shen Di Wan (DS, Cardiotonic Pill or Dantonic(R) Pill) is one of the most commonly used Chinese herbal formulations for treating patients with atherosclerotic disease in China and several Asian countries. We studied the effect of DS on platelet and leucocyte function and compared the effects with conventional antiplatelet agents, cangrelor (ADP P2Y(12) receptor antagonist) and aspirin (acetyl salicylic acid, ASA). Measurements were made by platelet aggregation (%) and activation (CD62P %), platelet-monocyte conjugate formation (P/M, CD42a median fluorescence, mf), platelet-neutrophil conjugate formation (P/N, mf), and leucocyte activation (CD11b median fluorescence on monocytes and neutrophils, mf) in response to 3.3 micromol/L adenosine diphosphate (ADP), 1.0 micromol/L platelet activating factor (PAF), 5.0 micromol/L adrenaline and 0.5 microg/mL collagen. We also evaluated the effect of its main component, water soluble extract of salvia miltiorrhiza (SME) on intracellular calcium mobilization in platelets triggered by 10 micromol/L ADP, 10 micromol/L PAF, 2 microg/mL collagen and 15 micromol/L thrombin receptor activating peptide (TRAP). Overall DS showed inhibition of platelet aggregation, platelet activation, platelet-leucocyte conjugate formation and leucocyte activation in response to all the agonists apart from adrenaline (all p < 0.01). DS showed inhibition of platelet aggregation and leucocyte activation equivalent to cangrelor 100 nmol/L and ASA 100 micromol/L. SME dose-dependently inhibited intracellular calcium mobilization in platelets following stimulation with all the platelet agonists with maximum effective at 0.36 mg/mL (all p < 0.01). When used at 0.18 mg/mL its inhibitory effect was equivalent to cangrelor and ASA. We conclude that DS is a potential inhibitor of both platelet and leucocyte activation.