Polyswitch lentivectors: "all-in-one" lentiviral vectors for drug-inducible gene expression, live selection, and recombination cloning

Lentiviral vectors are now widely considered one of the safest and most efficient tools for gene delivery and stable gene expression. Even though inducible gene expression cassettes are mandatory for many genetic engineering strategies, most current systems suffer from various issues, such as the requirement of two vectors, which decreases the overall efficiency of the transduction, leakiness and/or insufficient levels of activation of the inducible promoter, lack of selectable marker, low titers, or general issues associated with the cloning of large plasmids. In this article, we describe the design and functional characterization of a set of "all-in-one" multicistronic autoinducible lentivectors. They combine: (1) an optimized drug-inducible promoter; (2) a multicistronic strategy to express living color, selectable marker, and transactivator; and (3) acceptor sites for easy recombination cloning of genes of interest. These polyswitch lentivectors have good titers, very low basal activity, and reversible high induced activity, and can accept a growing number of genes already cloned in entry plasmids. These combined features make them a novel, powerful, and versatile tool for current and future genetic engineering approaches.     

Post-extinction fluoxetine treatment prevents stress-induced reemergence of extinguished fear

Rationale

The post-extinction exposure of rats to a sub-conditioning procedure (SCP; i.e., retraining with a shock intensity that is too weak to induce by itself significant fear conditioning) has been reported to provoke the reemergence of extinguished fear. This phenomenon can be prevented by chronic fluoxetine treatment.

Objectives

We sought to examine another potential inducer of fear reemergence, acute stress, in rats and determine whether fluoxetine prevents this phenomenon.

Methods

Because in previous studies fluoxetine was administered before extinction, we first analyzed its effect on the SCP-associated reemergence of auditory-cued conditioned fear in rats injected after extinction to avoid any interaction between fluoxetine and extinction learning. Next, we used the same protocol but replaced the SCP with acute stress.

Results

We found that the SCP and acute stress, which were carried out 3 weeks after fear extinction, similarly provoked the reemergence of extinguished fear in rats injected with vehicle during the 3-week period. In contrast, the animals treated with fluoxetine during this period behaved similarly to those not exposed to an inducer of fear reemergence.

Conclusions

Our data establish acute stress as an inducer of fear reemergence. The results provide further support for the hypothesis that fluoxetine interfered with mechanisms that reactivated extinguished fear, even when administered after fear extinction.     

Diagnostic performance and impact on patient management of <Superscript>68</Superscript>Ga-DOTA-TOC PET/CT for detecting osteomalacia-associated tumours

Purpose

Oncogenic osteomalacia is an endocrine disorder induced by small benign tumours (TIO) producing excessive fibroblast growth factor-23 (FGF23). The only way of curing oncogenic osteomalacia is surgical resection of the culprit TIO, which is extremely difficult to detect using conventional imaging modalities due to its small size and variable location in the body. Since TIO frequently overexpress somatostatin receptors, a clinical utility of SPECT or PET with radiolabelled somatostatin analogues has been reported. Among them, ⁶⁸Ga-DOTA-TOC has recently been granted a marketing authorization, facilitating its routine application. We report here the results of the first series evaluating the diagnostic performance of ⁶⁸Ga-DOTA-TOC PET/CT in detecting TIO and its impact on patient management.

Methods

⁶⁸Ga-DOTA-TOC PET/CT and clinical and imaging data from 15 patients with clinical and biochemical signs of oncogenic osteomalacia were retrospectively reviewed. The ⁶⁸Ga-DOTA-TOC PET/CT findings were compared with the results of post-surgical pathology and clinical and biochemical follow-up.

Results

⁶⁸Ga-DOTA-TOC PET/CT resulted in the detection of one focus suspicious for TIO in nine of 15 patients (60%), and a tumour was surgically removed in eight. Post-operative pathology confirmed a TIO in those eight patients whose symptoms diminished promptly and biochemical anomalies resolved. ⁶⁸Ga-DOTA-TOC PET/CT sensitivity, specificity and accuracy were 73%, 67% and 71%, respectively. ⁶⁸Ga-DOTA-TOC PET/CT findings affected patient management in 67% of cases. In particular, ⁶⁸Ga-DOTA-TOC PET/CT was able to detect the TIO with a negative or a false-positive result of a previous ¹¹¹In-pentetreotide SPECT/CT in 5/8 patients (63%) or a previous FDG PET/CT in 7/11 patients (64%). No close relationship was found between the positivity of ⁶⁸Ga-DOTA-TOC PET/CT and the serum level of a biochemical marker. However, a true-positive result of ⁶⁸Ga-DOTA-TOC PET/CT was obtained in only one patient with a non-elevated serum level of FGF23.

Conclusion

⁶⁸Ga-DOTA-TOC PET/CT is an accurate imaging modality in the detection of TIO; in particular, it is worthwhile after failure of somatostatin receptor SPECT(/CT) or FDG PET/CT.

     

超声乳化、囊袋内人工晶状体植入联合玻璃体切割术治疗增殖性糖尿病视网膜病变

目的:评价晶状体超声乳化、囊袋内人工晶状体植入联合玻璃体切割术治疗增殖性糖尿病视网膜病变的临床效果。方法:回顾性分析了33例(41眼)行晶状体超声乳化、后房型人工晶状体植入、玻璃体切割联合手术治疗增殖性糖尿病视网膜病变患者的病程、视力及手术并发症等临床资料。结果:41眼均为囊袋内植入人工晶状体,后囊膜完整。41眼中36眼(88%)视力提高,视力>0.05者占73%(30/41),>0.2者占46%(19/41),平均随访时间9mo。31眼术前未接受过眼内激光治疗,术中行全视网膜光凝。5眼术后玻璃体再次出血,1眼术后视网膜脱离复发,其中4眼再次行玻璃体视网膜手术。结论:晶状体超声乳化、囊袋内人工晶状体植入联合玻璃体切割术治疗增殖性糖尿病视网膜病变是安全有效的,有利于保持血—房水屏障,减少术后虹膜红变,早期恢复患者视力,避免再次白内障手术或二期人工晶状体植入。     

Sex matters: association between callous-unemotional traits and uncinate fasciculus microstructure in youths with conduct disorder

Brain Imaging and Behavior - Among youths with conduct disorder, those with callous-unemotional traits are at increased risk for persistent antisocial behaviour. Although callous-unemotional traits...     

Ueber Ependymveränderungen bei tuberculöser Meningitis

Ohne ZusammenfassungHierzu Taf. VI.     

Economic impact of clinical pharmaceutical activities in hospital wards: A systematic review

BackgroundThe positive impact of clinical pharmacy services (CPS) in improving clinical outcomes such as reduction of drug related problems is well demonstrated. Despite these results, the deployment of these activities is not systematically observed in the hospital setting.ObjectivesThis systematic review first aimed to describe existing evidence regarding economic evaluation of ward-based CPS focusing on the entire treatment of a patient in a hospital setting. Secondly, the quality of economic evaluations of existing evidence was assessed.MethodsA comprehensive literature search was performed in PubMed/Medline, Science Direct and the NHS Economic Evaluation databases from January 2000 to March 2019. English or French language articles describing an economic evaluation of ward-based CPS on inpatients in hospital settings were included. Articles not describing a single study, dealing with a CPS not considering the entire medication regimen of the patient or presenting both inpatient and outpatient CPS were excluded. Selected articles were analyzed according to Drummond's check-list for assessing economic evaluations.ResultsForty-one studies were included. About one third were American publications. CPS implemented in ICU represented about half of the selected articles. Pharmacist-to-bed ratios varied according to countries and care unit type with the most favorable ratios in ICU and in American studies. Cost-avoidance was mostly used to express economic impact and ranged from €1579 to €3,089 328. Studies yielding the greater economic impact were conducted in the USA with implementation of full-time equivalents pharmacists or establishing of collaborative practice agreements. Only 6 articles dealt correctly with at least 7 of the 10 Drummond's checklist assessment criteria.ConclusionThis review suggests that the existing evidence is not sufficient to conclude to a positive economic impact of CPS conducted according to clinical pharmacy guidelines. Funding resources, remuneration of clinical pharmacy activities and provision of standardized national clinical and economic databases appear to be essential evolutions to improve CPS development.