Quantification of diphtheria toxin mediated ADP-ribosylation in a solid-phase assay

BACKGROUND: Because of reduced vaccination programs, the number of diphtheria infections has increased in the last decade. Diphtheria toxin (DT) is expressed by Corynebacterium diphtheriae and is responsible for the lethality of diphtheria. DT inhibits cellular protein synthesis by ADP-ribosylation of the eukaryotic elongation factor 2 (eEF2). No in vitro system for the quantification of DT enzymatic activity exists. We developed a solid-phase assay for the specific detection of ADP-ribosylation by DT. METHODS: Solid phase-bound his-tag eEF2 is ADP-ribosylated by toxins using biotinylated NAD(+) as substrate, and the transferred biotinylated ADP-ribose is detected by streptavidin-peroxidase. DT enzymatic activity correlated with absorbance. We measured the amount of ADP-ribosylated eEF2 after precipitation with streptavidin-Sepharose. Quantification was done after Western blotting and detection with anti-his-tag antibody using an LAS-1000 System. RESULTS: The assay detected enzymatically active DT at 30 ng/L, equivalent to 5 mU/L ADP-ribosylating activity. Pseudomonas exotoxin A (PE) activity was also detected at 100 ng/L. We verified the assay with chimeric toxins composed of the catalytic domain of DT or PE and a tumor-specific ligand. These chimeric toxins revealed increased signals at 1000 ng/L. Heat-inactivated DT and cholera toxin that ADP-ribosylates G-proteins did not show any signal increase. CONCLUSIONS: The assay may be the basis for the development of a routine diagnostic assay for the detection of DT activity and highly specific inhibitors of DT.     

Appreciative inquiry for leading in complex systems: supporting the transformation of academic nursing culture

Increasingly complex environments in which nurse educators must function create distinct challenges for leaders in nursing education. Complexity is found in the presence of knowledge-driven economies, advancements in technology, and the blurring of campus boundaries created by online learning versus traditional classroom education. A dual bureaucracy of faculty and administration coexists in nursing education. The transformation of bureaucratic culture is a strategic challenge for academic leaders who strive to move dichotomous groups toward a collective vision of a preferred future. This article advocates for the affirmative administrative process of appreciative inquiry for academic nursing leadership, in nudging the dual bureaucracy toward transformational change. The intent and characteristics of appreciative inquiry are discussed, appreciative leadership strategies and actions are explained, methods for leading cultural paradigm shift are outlined, and an exemplar of the actualization of appreciative inquiry is presented.     

The effects of smoking on corneal endothelial cells: a cross-sectional study on a population from Isfahan,Iran

Purpose: The aim of this study is to compare the effect of smoking in corneal endothelial cell number and morphology by specular microscopy on a non-smoker population.

Methods: Our cross-sectional study was performed on 150 participants from a non-smoker population. Non-contact specular microscopy (Tomey Corporation Inc., Nagoya, Japan) was performed in the center of the cornea of all subjects. The cell density (CD), average cell size (AVG), percent of hexagonality (HEX%) and central corneal thickness (CCT) were calculated and compared in both groups.

Results: Totally, 76 eyes of 76 smokers and 74 eyes of 74 non-smokers were enrolled in the study from 2015 to 2016. The mean age of smokers and non-smokers were 48.61?±?17.04 and 46.39?±?13.02, respectively. The mean number of pack/year among the smokers was 17.36?±?14.68. Also, the mean values of AVG and CD were significantly different for these two groups (p?=?0.011 and p?=?0.039, respectively). Other corneal endothelial variables did not show a significant difference between smokers and non-smokers (p?>?0.05). However, smokers with severe nicotine dependency had significantly greater AVG and lower CD in comparison with the non-smokers (p?=?0.004 and p?=?0.013, respectively).

Conclusion: Our study showed that smoking can cause significant changes in some of the corneal endothelial variables, but not all of them.     

Pharmacovigilance workflow in Europe and Italy and pharmacovigilance terminology

The terminology used in pharmacovigilance can cause confusion, because there are similar terms that describe different phenomena (e.g. adverse reactions, adverse drug reactions, and side effects). Incorrect use of terminology can have negative effects on the reporting of adverse drug reactions and on the interpretation of these reports. To explain the most common terms used in pharmacovigilance, this article first describes the pharmacovigilance workflow process in the European Union and, as an example, in Italy. Then, the article reviews common pharmacovigilance terms.     

EU pharmacovigilance regulatory requirements of anticancer biosimilar monoclonal antibodies

An increasing number of innovative oncology monoclonal antibodies (mAbs) have been introduced into the global market, and biosimilar versions have now also been approved in Europe. Being complex to develop and difficult to manufacture, the biosimilar is a drug similar but not identical in physicochemical characteristics, efficacy, and safety to an original biological drug already approved in the European Union, for which marketing exclusivity rights have expired. Generally, the safety monitoring of biosimilars follows the same requirements that apply to all biologicals, even if specific pharmacovigilance measures exist and some of them are still being debated. The manufacturing process, immunogenicity, traceability, and extrapolation of indication are keywords which may impact on the achievement of additional knowledge about the safety of a biosimilar mAb. In this article, we aim to discuss elements that play a central role in the pharmacovigilance legislation of biosimilar mAbs.     

An historical overview over Pharmacovigilance

Pharmacovigilance started about 170 years ago, although it was not yet named as such at that time. It is structured activity in the professional health field, with important social and commercial implications aimed at monitoring the risk/benefit ratio of drugs, improving patient’s safety and the quality of life. In this commentary we report the milestones of pharmacovigilance up to the present day, in order to understand all the steps that have characterized the historical evolution; from the first reports, which were essentially letters or warnings sent by clinicians to publishers of important and famous scientific journals, up to today’s modern and ultra-structured electronic registries. The historical phases also help us to understand why pharmacovigilance helped us to achieve such important results for man’s health and for pharmacology itself, and to identify the challenges that await Pharmacovigilance in future years.     

Impact of emergency medicine faculty on door to thrombolytic time

The objective of this study was to determine the impact of Emergency Medicine (EM) faculty presence on timely thrombolytic therapy for acute myocardial infarction in the Emergency Department (ED). We performed a retrospective study of data regarding acute myocardial infarction patients in the ED of a large urban teaching hospital. Data were collected from January 1, 1998 to December 31, 1999 when EM faculty were not present in the ED and from January 1, 2001 to December 31, 2002 when they were. We compared median time from patient arrival to thrombolytics, percent of patients receiving thrombolytics within 30 min of arrival, and percent of patients with indications for primary revascularization who received it before and after EM faculty presence. The results indicate that EM faculty presence resulted in a decrease in median time from arrival to thrombolytic administration of 17 min (95% CI: 9, 28). Before EM faculty presence, the median time was 44 min as compared to 24 min post-EM faculty presence. Patients received thrombolytic therapy within 30 min 25.8% of the time before EM faculty presence as compared to 65.4% with EM faculty presence; an absolute increase of 39.6% (95% CI: 23.0%, 56.3%). Primary revascularization occurred in 56.9% of eligible patients pre-EM faculty presence and 81.4% post-EM faculty presence; an increase of 24.5% (95% CI: 13.6%, 35.4%). We conclude that the introduction of Emergency Medicine faculty significantly improved the quality of care for acute myocardial infarction patients in a large urban Emergency Department.     

A retrospective case–control study of gastrostomy use in children undergoing hematopoietic cell transplantation

Background

Maintaining a good nutritional status during the hematopoietic cell transplantation (HCT) procedure is challenging in the pediatric population.

Methods

In a multicentric retrospective study, we compared the outcome of nutritional status and HCT-related parameters in 227 pediatric patients during and after HCT between 2005 and 2015. 112 patients received a gastrostomy before the start of HCT (GS group), and 115 did not receive a gastrostomy (NGS). Data collection was performed at HCT, 3, 6, and 12 months post-HCT.

Results

At time point of HCT the Standard Deviation Score (SDS) of weight was 0.17 in the NGS group, and 0.71 in the GS group (p = .01) Patients in the NGS group lost more weight during the first 3 months after HCT than patients in the GS group. At 12 months, patients in the NGS remained at a lower weight, while patients in the GS group slightly increased their weight. There were no differences between the groups in the incidence of acute graft-versus-host-disease (GvHD), overall survival, and non-relapse mortality. However, the number of febrile episodes requiring intravenous treatment with antibiotics, was higher in the GS group as compared to the NGS group, during the first 3 months post-HCT (p < .001).

Conclusions

Our results indicate that gastrostomy can be utilized in children undergoing HCT without any negative effects on mortality. Therefore, the use of a gastrostomy appears to be a safe option to maintain a good nutritional status during the HCT procedure.     

Local treatment of rhabdomyosarcoma of the female genital tract: Expert consensus from the Children's Oncology Group,the European Soft-Tissue Sarcoma Group,and the Cooperative Weichteilsarkom Studiengruppe

The International Soft-Tissue Sarcoma Consortium (INSTRuCT) was founded as an international collaboration between different pediatric soft-tissue sarcoma cooperative groups (Children's Oncology Group, European Pediatric Soft-Tissue Sarcoma Group, and Cooperative Weichteilsarkom Studiengruppe). Besides other tasks, a major goal of INSTRuCT is to develop consensus expert opinions for best clinical treatment. This consensus paper for patients with rhabdomyosarcoma of the female genital tract (FGU-RMS) provides treatment recommendations for local treatment, long-term follow-up, and fertility preservation. Therefore, a review of the current literature was combined with recommendations of the treatment protocols of the appropriate clinical trials. Additionally, opinions of international FGU-RMS experts were incorporated into recommendations. Results were that the prognosis of FGU-RMS is favorable with an excellent response to chemotherapy. Initial complete surgical resection is not indicated, but diagnosis should be established properly. In patients with tumors localized at the vagina or cervix demonstrating incomplete response after induction chemotherapy, local radiotherapy (brachytherapy) should be carried out. In patients with persistent tumors at the corpus uteri, hysterectomy should be performed. Fertility preservation should be considered in all patients. In conclusion, for the first time, an international consensus for the treatment of FGU-RMS patients could be achieved, which will help to harmonize the treatment of these patients in different study groups.     

The effects of saffron supplementation on the measures of renal function indicators: a systematic review and meta-analysis

International Urology and Nephrology - Saffron (Crocus sativus L.) has been proposed as a potential agent to improve renal function in animal studies. But, due to insufficient evidence in human...